spex handbook 2011

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Handbook of SAMPLE PREPARATION and HANDLING

Eleventh Edition Contributing EditorsRalph H. Obenauf, Ph.D.John MartinRichard BostwickTim Osborn-JonesLea Anderson-Smith, Ph.D.Ben MurrayKeith TuckerDave SamuelsSandy ManganSteve SiniscalcoSarah Geiger SPEX SamplePrep, LLC15 Liberty St.Metuchen, NJ 08840Phone: (732) 623-0465Fax : (732) 906-2492Email : [email protected]: www.spexsampleprep.com European Division:SPEX Ceriprep Ltd.2 Dalston GardensStanmoreMiddlesex HA7 1BQUnited KingdomPhone: +44 (0) 208 204 6656Fax : +44 (0) 208 204 6654Email: [email protected]: www.spexsampleprep.com

Order Online 24/7: www.spexsampleprep.com2

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Introduction

SPEX SamplePrep, LLC has been providing superior sample preparation equipment and supplies since 1954. Our mission is to provide quality products backed by exceptional service and expertise. SPEX SamplePrep equipment is used to prepare samples for a wide range of analytical technologies including XRF, AA, and ICP. Much of our equipment is also used for cutting-edge research in pharmaceuticals, superconductors, polymers, forensics and genetics.

Our Products

SPEX SamplePrep products include:

• Freezer/Mill® – Uniquely designed to cryogenically grind samples normally considered difficult or impossible to pulverize at ambient temperature

• Geno/Grinder® – Used for high-throughput cell disruption and tissue homogenization

• Mixer/Mill® – High-energy ball mills for pulverizing brittle materials, mixing powders and emulsions, mechanical alloying, and nanomilling

• Shatterbox® – Swing mills used for rapidly pulverizing brittle materials such as cement, slag, ores, ceramics, etc.

• Katanax® Electric Fusion Fluxers – Automated electric fluxers for borate fusions of cement, ores, ceramics and other materials for analysis by XRF, AA, ICP and wet chemistry

• X-Press® and Bench-Press® – Laboratory presses ideal for pressing sample disks for XRF, IR, and OES

• XRF Accessories – Spec-Caps® for supporting pressed disks, disposable X-Cells® and window film for liquid samples, PrepAid® Binders and Grinding Aids

Our Quality

SPEX SamplePrep equipment has become the industry standard for reliability and durability in the laboratory and our products have been used in landmark studies such as those of NASA’s moon rocks and the identification of the remains of Czar Nicholas II and his family. Our expertise and products can help you achieve accurate, consistent results by ensuring reproducible samples, analysis after analysis. Let our five decades of sample preparation experience work to your advantage in the lab.

Our Customers

For the past five decades, SPEX SamplePrep’s customers have relied on our unique knowledge and expertise to select the proper products for their sample preparation needs. Our exceptional sales and service teams recommend and support reliable sample preparation equipment for the following fields:

• Cement Manufacturing • Pharmaceuticals and Drug Testing • Mining • Cosmetics • Geology and Mineralogy • Forensics and Crime Scene Investigation • Polymers • Environmental Protection and Remediation • RoHS/WEEE • Food and Agriculture • Medicine and Genetics

How To Order

By MailMail your orders to: SPEX SamplePrep, LLC 15 Liberty St Metuchen, NJ 08840

By Phone(732) 623-0465 – US+44 (0) 208 204 6656 - UK

By Fax(732) 906-2492 – US+44 (0) 208 204 6654 – UK

Via [email protected] – [email protected] - UK

By Webwww.spexsampleprep.com


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We are pleased to announce the launch of our brand new website to support the growing demand for our products and services. Our website offers a complete e-commerce solution to aid in your purchase of laboratory equipment and accessories for every sample size and application. Registering on our website is a simple process and gives you access to:

• Product Offering and Pricing

• Online Ordering

• Order Status & Order History - Check your complete order history

• Live Chat - For all the questions you need answered immediately, our sales staff can assist you

• Newsletters - Register for our newsletters to receive all the latest updates on SPEX SamplePrep equipment and for exclusive offers and promotions

Log on today to find out why SPEX SamplePrep is the better solution:

www.spexsampleprep.com

When placing your order please include:

SPEX SamplePrep Account Number (if known)Purchase Order NumberBilling AddressShipping AddressCatalog number (in full as shown herein)Quantity (in units, not pieces)Item DescriptionPreferred Shipping MethodPayment MethodTax Exempt Number (if applicable)Copy of Tax Exempt Certificate (if applicable)

Distributors

SPEX SamplePrep distributes products through a global distribution network. To find a distributor in your country please visit:

www.spexsampleprep.com/purchase-options/distributors.aspx

Alternatively you can contact our main office for more information on distributors in your country.

Prices and Quotations

Prices are subject to change without notice. We will gladly quote current prices on request. Verbal and written quotes are valid for 30 days unless otherwise stated. Acceptance by the seller is expressly limited to the Standard Terms of SPEX SamplePrep, LLC.

Confirming Orders

Mark confirming orders clearly. Failure to do so will result in a 25% restocking charge for duplicate shipments.

Domestic Orders (US)

PaymentPayment terms are net 30, days based on credit approval. If you do not have an established account with SPEX SamplePrep, LLC, you may provide credit information, three commercial references, Dun and Bradstreet Number, or remit payment including freight charges with order. Payment may also be made by credit card (Visa, MasterCard or American Express), or wire transfer.

ShippingOur shipping terms are FCA Ex Works Metuchen, NJ. Any other terms are by special arrangement only. Most items are shipped by UPS or motor freight, depending on the size and weight of the shipment. Various express/overnight services are also available. All shipping charges are prepaid and the cost is added to the invoice. If you are sending payment with the order, consult us first for shipping charges.

Minimum OrdersThe minimum order is $100. Replacement parts and repair charges are excluded from this minimum.

Export Orders

SPEX SamplePrep maintains authorized dealers in many countries around the world. In Europe we are represented by our subsidiary, SPEX CertiPrep, Ltd. To find an authorized dealer in your country visit our website or contact SPEX SamplePrep via email at [email protected].

Orders from countries without authorized SPEX SamplePrep dealers may be placed directly with SPEX SamplePrep, 15 Liberty St., Metuchen, NJ 08840. However, export ordering terms, instructions, and quotations must be obtained in advance. Orders received without prior agreement may be returned to the purchaser.


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PaymentPrepayment of merchandise cost and shipping charges is required for all export orders. Payment may be made by check, wire transfer, letter of credit, or credit card (Visa, MasterCard, American Express). Minimum order for letter of credit payments is $2,500.00. All banking charges must be to buyer’s account. Any letter of credit that states “banking charges are for the account of the beneficiary” WILL NOT BE ACCEPTED.

Minimum OrdersThe minimum order is $100. Replacement parts and repair charges are excluded from this minimum.

Warranty

SPEX SamplePrep LLC guarantees its products against defects of materials and workmanship for one year from the date of original shipment. Repairs and replacements made under warranty are guaranteed for the remaining original warranty period. The warranty excludes wear parts: parts that wear out through use and have to be replaced periodically for proper operation. Wear parts include not only gaskets, drive belts, grinding media, and the like, but also pneumatic cylinders, the Geno/Grinder ball slide, the Freezer/Mill coil, and the Mixer/Mill clamp retaining spring assembly. Also excluded from the warranty are grinding vials and containers made from tungsten carbide, alumina ceramic, zirconia ceramic, silicon nitride, and agate. The customer pays return freight for warranty claims, but SPEX SamplePrep will pay return freight to the customer if the warranty claim is valid. SPEX SamplePrep reserves the right to judge whether a malfunction is due to defects in materials or workmanship, or to wear, negligence or misuse.

All Katanax instruments have been carefully inspected and tested before shipping and are warranted to be free from defects in workmanship and material for a period of one year from date of shipment. Hard ceramic parts may exhibit small cracks developed under heat, and will not be considered defective unless this situation impairs functionality of the instruments. Heating elements are warranted for a period of six (6) months.

To Arrange a Return Shipment

We want you to be happy with your purchase from SPEX SamplePrep. Please bring any problem to our attention, but DO NOT RETURN any item before contacting us for a RMA (Returned Materials Authorization) number, and instructions. Unauthorized returns will be refused. Cost for all return transportation is the responsibility of the customer. Credit for returned merchandise is issued only after the goods have been received and inspected, and returns are subject to a 25% restocking fee up to a maximum of $200.00. Address all shipments to SPEX SamplePrep, LLC, 15 Liberty St., Metuchen NJ 08840; always include the RMA number.

Service

The majority of our products are designed and built in our factory in Metuchen, New Jersey. The SPEX SamplePrep Service Department is an integral part of our operation, and can be counted on to maintain, upgrade, repair, or rebuild SPEX SamplePrep equipment with factory parts and the necessary expertise. Our trained service technicians can also advise you on the most likely cause of a problem. Repairs are subject to a minimum two-hour labor charge, and prices are subject to change without notice. Out-of-warranty repairs and parts are guaranteed for 30 days. Before returning equipment for service, contact our technicians for an RMA number and cost estimate, and include a purchase order with the return. Address returns to our 15 Liberty St. address as described above. The customer pays freight both ways for equipment to be serviced.

Product Specifications

Every effort has been made to provide complete and accurate product information in this catalog. However, specifications are subject to change without notice, and changes may be made from time to time to improve the performance of our products.

Precautions

SPEX SamplePrep products are not for any household application. Our acceptance of a purchase order is made with the assumption that our products will be used only by qualified individuals who are trained in appropriate procedures. Users are responsible for knowledge and understanding of the potential hazards of the material with which they are working. SPEX SamplePrep does not recommend unattended operation of any of our laboratory equipment.


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Table of Contents

Sample Preparation Techniques

Sample preparation techniques are the building blocks used to prepare samples for specific scientific analyses. Good techniques are the foundation to ensuring high quality, reliable analytical results. Our vast line of equipment and accessories are used for a variety of common techniques as well as specialties, such as cryogenic grinding. Below is a short description of each technique along with some insight into the use of our equipment.

Grinding & Pulverizing

SPEX SamplePrep’s laboratory mills and grinders are capable of pulverizing samples in both ambient and cryogenic environments. This versatility allows for grinding of a wide variety of samples, including metals, rocks, plastics, plant and animal tissue, bone, pharmaceuticals, and more.

Mixing & Blending

The non-homogeneity of materials in the real world presents a serious sampling problem for scientists. SPEX SamplePrep’s mills are effective at mixing and blending dry materials such as pigments, powders, pharmaceuticals, and cosmetics as well as samples that require mixing for effective extraction with a solvent or buffer.

Tissue Homgenization /Cell Lysis

For Biotechnology and Forensic applications, effective cell disruption and DNA/RNA extraction are critical. SPEX SamplePrep’s Geno/Grinder and Freezer/Mills aid scientists by providing high-throughput sample preparation solutions at both ambient and cryogenic temperatures.

Pressing & Pelletizing

Analytical spectroscopic methods such as XRF, OES, and IR often require samples in the form of a round, flat-surfaced disc. SPEX SamplePrep’s laboratory presses are ideal for preparing sample discs for these methods. In addition, our automatic and air-actuated presses are designed to fit on the bench top, requiring less space than standard floor model presses.

Borate Fusion

For samples that are difficult to prepare as homogeneous pressed powders (cement), difficult to dissolve in acid (zirconia and alumina), or both (metal ores and silicate rocks), borate fusions are widely used. SPEX SamplePrep offers the full line of Katanax Electric Fusion Fluxers & SPEX CertiPrep Fusion Flux for preparation of samples for XRF, AA, and ICP.

Table of Contents

Introduction

How to Use This Handbook. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15Sample Preparation Techniques Reference Chart . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

SECTION 1 :: Products and Accessories

Chapter 1SPEX SamplePrep 2010 Geno/Grinder® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19Clamps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19Grinding Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19Titer Plates and Cap-Mats . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22Vials and Vial Sets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23Racks and Holders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27Kryo-Tech® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28Cryo-Blocks. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Chapter 2SPEX SamplePrep 6970EFM Enclosed Freezer/Mill® . . . . . . . . . . . . . . . . . . . . . . . . 30SPEX SamplePrep 6870 Freezer/Mill®. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31SPEX SamplePrep 6770 Freezer/Mill®. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33Vials & Vial Sets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33Vial Racks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35Extractors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35Sample Adapters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36Accessory Packages & Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37Liquid Nitrogen Handling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

Chapter 3SPEX SamplePrep 8000D Dual Mixer/Mill®. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40SPEX SamplePrep 8000M Mixer/Mill®. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41


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Table of Contents

SPEX SamplePrep 5100 Mixer/Mill® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43Vials & Vial Sets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43Small Vials and Vial Sets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43Large Vials and Vial Sets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44Comparison of Grinding Vials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47Grinding Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48Sample Adapters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49SPEX SamplePrep PrepAid® Grinding Aids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

Chapter 4SPEX SamplePrep 8530 Enclosed Shatterbox® . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 SPEX SamplePrep 8500 Shatterbox® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52Grinding Dishes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52Racks and Holders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54PrepAid® Grinding Aids & Sample Binders. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54

Chapter 5SPEX SamplePrep 3635 Automated X-Press® . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55SPEX SamplePrep 3628 Air-Actuated Bench-Press® . . . . . . . . . . . . . . . . . . . . . . . . 563621 SPEX SamplePrep-Carver Model C . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 573626 SPEX SamplePrep-Carver 12-ton Press . . . . . . . . . . . . . . . . . . . . . . . . . . . . 583622 SPEX SamplePrep-Carver Model M . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59Pellet Die Sets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59Replacement Pellets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60Sleeve-and-Plunger Sets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60Spec-Caps® for Reinforcing XRF Sample Disks. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61SPEX SamplePrep PrepAid® Binders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62

Chapter 6Katanax® K1 Prime Automated Electric Fluxer . . . . . . . . . . . . . . . . . . . . . . . . . . . 64Katanax® K2 Prime Automated Electric Fluxer . . . . . . . . . . . . . . . . . . . . . . . . . . . 65Crucibles and Molds for Katanax K1 and K2 Automated Electric Fluxers . . . . . . . . . . . . . . . . . . 66Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66Crucibles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66Racks & Holders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67Accessory Packages & Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

Platinumware, Labware & Handling Tools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67Molds & Mold Holders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67Mold Holders for the K1 Prime Automated Electric Fluxer. . . . . . . . . . . . . . . . . . . . . . . . . . . 68Mold Holders and Plates for the K2 Prime Automated Electric Fluxer . . . . . . . . . . . . . . . . . . . . 68Graphite Crucibles for muffle furnaces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69Fusion Flux and Additives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

Chapter 7XRF Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74Spec-Caps® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74PrepAid Binders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75X-Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76Thin Film . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78Certified Reference Materials for Organic and Inorganic Spectroscopy and Chromatography . . . . . . 81Services available from the CRM Division . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81Products available from the CRM Division . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82

SECTION 2 :: Technical Information

Chapter 1Tissue Homogenization / Cell Lysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86Bead Milling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86Cryogenic Grinding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

Chapter 2Grinding & Pulverizing / Mixing & Blending. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88When to Pulverize . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88How to Select a Laboratory Mill and Grinding Container . . . . . . . . . . . . . . . . . . . . . . . . . . . 88SPEX SamplePrep Container Materials Selection Chart . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90Composition of Standard Steels Used in SPEX SamplePrep Equipment & Accessories. . . . . . . . . . . 91Key Points About Cryogenic Grinding. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92Limits of Cryogenic Grinding. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93Guide to Freezer/Mill Sample Capacity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93Cryogenic Grinding for RoHS/WEEE. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94Grinding Tests with the Freezer/Mill . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96Key Points About Grinding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97Grinding Container Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97


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Table of Contents

Contamination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98Grinding Aids. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98Cleaning Containers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99Respiritory Protection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100Infrared Mulls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100Pelletizing Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100Mechanical Alloying. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100SPEX SamplePrep Mixer/Mills . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102Pulverizing and Blending Grinding Tests with 8000 Series Mixer/Mills . . . . . . . . . . . . . . . . . . 105SPEX SamplePrep Shatterbox . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107Grinding Tests with the 8501 and 8504 Grinding Containers . . . . . . . . . . . . . . . . . . . . . . . . 109

Chapter 3Pressing and Pelletizing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110Preparing Powder Samples for XRF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110Particle Size Effects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111Pressing Sample Disks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113Technique for Pressing a Sample Disk . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114The Use of the Vacuum in Pellet Pressing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114Set-Up and Loading of the SPEX SamplePrep Evacuable Pellet Die . . . . . . . . . . . . . . . . . . . . 115Care and Maintenance of the SPEX SamplePrep Evacuable Pellet Die . . . . . . . . . . . . . . . . . . . 115Proper Use of the SPEX SamplePrep Spec-Cap . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116Binders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117Sleeve-and-Plunger Technique: Binder as Sample Disk Matrix . . . . . . . . . . . . . . . . . . . . . . . 119Sample Disk Removal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119Anti-Sticking Agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120Sample Disk Handling and Care . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121Trouble Shooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122Selecting a Pellet Die . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123

Chapter 4Fusion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124When to Employ Borate Fusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124How to Prepare a Fused Sample. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126

How to Select a Fusion Flux . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126Additives for Fusions: Non-Wetting Agents, Fluidizers, and Oxidizers . . . . . . . . . . . . . . . . . . . 127Graphite Crucibles and Handling Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128

Chapter 5XRF Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130Reinforcing XRF Pellets with Spec-Caps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130X-Cells for XRF Liquid Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130Bubble-Free Cell Insert . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131Selecting Thin Film Windows for XRF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132Thin Film Transmission Characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134Chemical Resistance of Kapton®. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135Chemical Resistance of Mylar® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136Chemical Resistance of Polypropylene . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137Chemical Elements and Their Symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139

SECTION 3 :: Application Notes

Geno Grinder . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142SP001: Cell Disruption . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142SP011: DNA Extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143SP013: Grinding / Disruption . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 145SP014: RNA Extraction from Aspergillus parasiticus mycelium . . . . . . . . . . . . . . . . . . . . . . 146SP016: Lysing / Homogenization. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 148SP017: Lysis Time and Other Variables on DNA Extraction . . . . . . . . . . . . . . . . . . . . . . . . . 149SP018: High-Throughput Disruption of Yeast in a 96-Well Format . . . . . . . . . . . . . . . . . . . . 151SP019: Comparison of Methods for the Isolation of DNA from Soybeans . . . . . . . . . . . . . . . . 153SP020: Lysing of Bacterial Cells in the Geno/Grinder . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156SP021: Extraction of RNA/cDNA and Genomic DNA from Tissue with Real-Time PCR . . . . . . . . . 158 SP022: The Benefits of the Geno/Grinder for Pesticide Residue Analysis. . . . . . . . . . . . . . . . . 162SP023: Comparison of Pesticide Extraction in Agricultural Products . . . . . . . . . . . . . . . . . . . 172SP024: Pesticide Anaylsis: Standard QuEChERS vs Modified Method . . . . . . . . . . . . . . . . . . . 175SP025: Lysing / Homogenization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183


Table of Contents


Introduc tion

Freezer Mill . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184SP002: Ginding/Homogenization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184HT 001: Grinding Bone in the Freezer/Mill . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185SP005: DNA/RNA Extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187SP006: Milling/Blending (Homogenizing) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189SP007: Grinding/Homogenization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190SP008: Structural Changes in Polymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192SP009: Reduction/Homogenization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193SP015: RNA Extraction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194

Mixer Mill . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196SP003: Nano Technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196HT002: Crude Fat (Hexane Extractables) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 198

Katanax . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 200SP026: Preparation and XRF Analysis of Fly Ash Fused Beads . . . . . . . . . . . . . . . . . . . . . . . 200

Appendix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203Periodic table. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203XRF Sample Preparation: Selected References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205Trademark References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 206Questionnaire for test grinding of Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207 Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208Subject Index. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208Product Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 216

How to Use the SPEX SamplePrep Handbook of Sample Preparation and Handling

The SPEX SamplePrep Handbook can help you get the most from your analyses by increasing the reliability and reproducibility of your prepared samples. It suggests how the most heterogeneous, unmanageable materials can be transformed into samples ideal for your analysis, whether as fine, homogeneous powders, compressed powder sample disks, fused glass disks, or solutions. There are techniques for avoiding sample contamination, inhibiting caking during pulverization, protecting sample disks against breakage, and copying with many other sample preparation difficulties. The Handbook also includes a complete listing of reliable sample preparation products to make the analyst’s life easier.

The Handbook also includes a complete listing of reliable sample preparation products to make the analyst’s life easier.

The Handbook is divided into 3 sections:

• Section 1 - Product information such as descriptions and catalog numbers for ordering

• Section 2 - Technical information associated with these products along with sample preparation techniques and application

• Section 3- Our Application Notes library regarding the use of our equipment for your specific application

Each chapter within these sections pertains to a particular product, as well as the sample preparation technique as it relates to a particular product. Margin tabs have been included to differentiate these areas. The sample preparation techniques are discussed in further detail at the end of this introduction.

Procedural tips and applications-oriented product descriptions are supported by photographs, diagrams, and product comparison charts.

A subject index (pg. 208) makes it easy to locate information about particular products, samples, sample preparation techniques, or analytical methods. A numerical product index (pg. 216) helps you find product descriptions for SPEX SamplePrep laboratory equipment and supplies by model or catalog number.

Order Online 24/7: www.spexsampleprep.com16 17

Products& Accessories

Section 1

Would you like to test our sample preparation equipment with your own samples?

Now You Can With SPEX SamplePrep’s Demo ProgramContact SPEX SamplePrep to arrange a test grind in our labs by our experts. Or try out our sample

preparation equipment in your laboratory for one week.

Best of all, our demo program is complimentary!

You only pay shipping and handling to and from your facility. Once you try our quality products you’ll ask yourself how you ever lived without them.

Products Available For Demo:

Katanax K1 Prime Electric Fluxer

8000D & 8000M Mixer/Mill

6770/6870/6970EFM Freezer/Mill

2010 Geno/Grinder

8530 Shatterbox

3635 X-Press

Contact (732) 623-0465 Email: [email protected]

Katanax K1 Prime is only available for local demos. We recommend test samples be sent to us directly to be processed in our demo lab.


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SPEX SamplePrep 2010 Geno/Grinder®

2010-115 Geno/Grinder – For 115V/60Hz operation2010-230 Geno/Grinder – For 230V/50Hz operation, CE Approved

• Versatile high-throughput plant and animal tissue homogenizer/cell lyser with a unique, vigorous, vertical pulverizing motion

• Handles two standard deep-well titer plates or multiple vials from 0.6 to 50 mL

• Optional large clamp assembly (2199) enables processing of up to four deep-well titer plates or sixteen 50 mL vials simultaneously

• Controls allow the operator to set the run time in 5 second increments and precise vertical grinding speed between 500 and 1750 strokes per minute (SPM)

• Sample preparation is completed in as little as 2 minutes with the addition of 1-2 grinding balls to the sample with a buffer solution

• Safety interlocks and pneumatic cylinder stabilize the lid for safe operation

• Supplied with height-adjustment spacers to handle titer plates, racks, vials, and other configurations up to 2¼ in. height

• LCD screen displays the full timer setting along with the time, in minutes and seconds, remaining in the grinding cycle

Typical Samples and Applications

• Plant materials such as seeds, stems, roots, leaves, fruits, vegetables

• Animal tissue

• Isolation of nucleic acids (DNA/RNA), proteins, enzymes, bacteria, and yeast

• Extraction of pesticide residue using QuEChERS Method

• Extraction of oil from fish and seafood.

Specifications:

Dimensions: 25 ¼ in. (64 cm) x 15 in. (36 cm) x 20½ in. (52 cm)

Weight: 87 lbs. (39 kg) net

Voltage: 115V/60Hz or 230V/50Hz

Power Cord: 3-prong standard North American plug for 115V/60Hz version 2-prong European CEE 7/7 plug for 230V/50Hz version

Accessories

ClampsThe 2010 Geno/Grinder is supplied with the Standard Size Clamp Assembly, which is designed to accommodate two standard deep-well titer plates or a variety of small vials with a maximum height of 2¼ in (57 mm). For taller vials or to stack titer plates, the Large Capacity Clamp Assembly is available

2199 Large Capacity Clamp AssemblyAn oversized clamp that accommodates centrifuge tubes or a double load of titer plates. Designed to hold up to sixteen standard 50 mL centrifuge tubes (115 mm length) with 2196-16-PE Holder, or twenty-four standard 15 mL centrifuge tubes (119 mm length) with 2197 Holder. Clamp also holds four deep-well titer plates, stacked two per side. Rubber mat for positioning between stacked titer plates is included.

Grinding MediaGrinding Balls & Magnets Stainless steel balls are useful for grinding large or tough samples. The choice of ball size should be determined based on the sample material and the size of the grinding vial. After use, the grinding balls can be discarded or cleaned for re-use.

2100 Grinding Ball Dispenser Simultaneously dispenses one 5/32 in. (4 mm) steel grinding ball (2150) into each well of a standard 96-well titer plate, including 2200 and 2210 titer plates.

2110M 24-Pin Magnet24 magnet-tipped pins on plate with handle. Inserts or removes 3/8 in. (9.5 mm) steel grinding balls (2155) from titer plates and vial sets. Works with 2230 titer plate, 2240-PC and 2240-PE Vial Sets, and 3116PC vials in 2190 Vial Rack. Used with 2110S Separator.

2110S Separator for 24-Pin MagnetSeparates 3/8 in. (9.5 mm) steel grinding balls (2155) from 2110M 24-pin magnet. Drops balls into 2230 24-well titer plates, 2240-PC and 2240-PE Vial Sets, or removes balls from magnets after balls are pulled out of wells or vials.

Patented and Patent Pending


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2110M-12 Pack of 12 Magnetic TipsEach Magnetic Tip is a micropipette tip with a strong magnet. Converts micropipette dispensers into pin magnets for removing 2150 grinding balls from 96-well titer plates. Pack of 12 Magnetic Tips.

2110M-96 Pack of 96 Magnetic TipsEach Magnetic Tip is a micropipette tip with a strong magnet. Converts micropipette dispensers into pin magnets for removing 2150 grinding balls from 96-well titer plates. Pack of 96 Magnetic Tips.

2150 Grinding Balls, 5/32 in. (4 mm)Made of 440C stainless steel. Used with 2100 Grinding Ball Dispenser and all 96-well titer plates. Sold in bags of 5,000.

2155 Grinding Balls, 3/8 in. (9.5 mm)Made of 440C stainless steel. For use in 2230 titer plate or in 2190 Vial Rack with 3116PC, 2241-PC and 2241-PE Vials to grind extra large or tough samples. Sold in bags of 100.

2156 Grinding Balls, 7/16 in. (11 mm)Made of 440C stainless steel. For grinding large or tough samples in 2230 titer plates and 2250 Vial Sets. Sold in bags of 100.

Grinding BeadsMolecular Biology Grade Grinding Beads are treated to inactivate contaminating enzymes and have been tested accordingly. Low Binding Grinding Beads are coated to reduce non-specific binding of nucleic acids and proteins and are used for lysing dilute samples of cells. Acid Washed Grinding Beads are treated to remove fine particles and contaminants. They are suitable for most basic applications. SPEX SamplePrep offers all three grades in a variety of sizes.

2160 Silica Grinding Beads, 800-1000 µmAcid washed silica beads, size ranges from 800-1000 µm, 200 gram bottle.

2161 Silica Grinding Beads, Molecular Biology Grade, 800-1000 µmAcid washed RNase/DNase-free treated silica beads, size ranges from 800-1000 µm, 200 gram bottle.

2162 Low Binding Silica Beads, 800 µmAcid washed and chemically treated to keep samples from binding to titer plate wells, 200 gram bottle.

2165 Silica Grinding Beads, 400-600 µmAcid washed silica beads, size ranges from 400-600 µm, 200 gram bottle.

2166 Silica Grinding Beads, Molecular Biology Grade, 400-600 µmAcid washed RNase/DNase-free treated silica beads, size ranges from 400-600 µm, 200 gram bottle.



2170 Silica Grinding Resin, 100-400 µmAcid washed irregular silica resin, size ranges from 100-400 µm, 200 gram bottle.

2171 Silica Grinding Resin, Molecular Biology Grade, 100-400 µmAcid washed RNase/DNase-free treated irregular silica resin, size ranges from 100-400 µm, 200 gram bottle.

2180 Zirconium Grinding Beads, Molecular Biology Grade, 200-400 µmAcid washed RNase/DNase-free treated zirconium beads, size ranges from 200-400 µm, 250 gram bottle.

2181 Low Binding Zirconium Beads, 100 µmAcid washed and chemically treated to keep samples from binding to titer plate wells, 250 gram bottle.

2182 Low Binding Zirconium Beads, 200 µmAcid washed and chemically treated to keep samples from binding to titer plate wells, 250 gram bottle.

Grinding Cylinders Our ceramic cylinders are chemically inert and are useful when mixing samples for extraction methods such as pesticide residue analysis and extraction of oil from fish and seafood. The angle-cut ends help the cylinders to shear the sample matrix during processing, resulting in a thorough extraction of analyte into the solvent. Typically, 2 cylinders are used per sample vial.

2183 Ceramic Grinding Cylinder, 3/8 in. x 7/8 in.Large size ceramic grinding cylinder with angle-cut ends for use with standard 50 mL centrifuge tubes. Sold in bags of 100.

2184 Ceramic Grinding Cylinder, 5/16 in. x 5/8 in.Medium size ceramic grinding cylinder with angle-cut ends for use with standard 15 mL centrifuge tubes. Sold in bags of 100.

2185 Ceramic Grinding Cylinder, 5/32 in. x 5/16 in.Small size ceramic grinding cylinder with angle-cut ends for use with standard 5 or 15 mL centrifuge tubes. Sold in bags of 100.


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Titer Plates and Cap-Mats

While most titer plates can be used in the Geno/Grinder, these sturdy titer plates have been tested extensively with various samples, and resist perforation by steel grinding balls even at high clamp speeds. They can be used for many applications including sample libraries, mother-to-daughter automated plate pipetting, large sample dilutions and cell suspensions. Suitable for RNA extraction when used with 2600 Cryo-Station. Titer plates are made of polypropylene, with alphanumeric marks for well identification. Sold singly or in cases of 100.

Cap-Mats seal titer plate wells, preventing spills and well-to-well contamination. Rugged silicone rubber, may be sterilized and re-used. Sold singly or in cases of 10.

2200-100 96-Well Titer Plate, Square Wells, Case of 100Square 2.4 mL wells with a working capacity of 2.0 mL. Case of 100 titer plates.

2200 96-Well Titer Plate, Square WellsSame as above, but a single titer plate.

2201-10 Cap-Mats for 2200 Titer Plates, Case of 10Sealing mats for the 2200 Titer Plates, above. Case of 10 Cap-Mats.

2201 Cap-Mat for 2200 Titer PlateSame as above, but a single Cap-Mat.

2210-100 96-Well Titer Plate, Round Wells, Case of 100Round 2.0 mL wells, rugged polypropylene, alphanumeric marks for well identification. For use with the 2650 Cryo-Block, which is inserted in gaps between wells to keep plates and samples chilled to preserve RNA for extraction.

2210 96-Well Titer Plate, Round Wells Same as above, but a single titer plate.

2211-10 Cap-Mat for 2210 Titer Plate, Case of 10Sealing mats for 2210 Titer Plates. Tough silicone rubber, prevents leaks and cross-well contamination. Can be sterilized and re-used. Sold in cases of 10 mats.


2220-100 48-Well Titer Plate, Case of 100Rectangular 5 mL wells with a working capacity of 2 mL per well. Use with 2221 Cap-Mat and multiple 5/32 in. (4 mm) steel grinding balls (2150) per well. Case of 100 titer plates.

2220 48-Well Titer PlateSame as above, but a single titer plate.

2221-10 Cap-Mat for 2220 Titer Plate, Case of 10Cap-Mats seal 2220 48-Well Titer Plates. Case of 10 Cap-Mats.


2230-100 24-Well Titer Plate, Case of 100Square 10 mL wells with a working capacity of 4 mL per well. Use with 2231 Cap-Mat, and one 3/8 in. (9.5 mm) steel grinding ball (2155) or 7/16 in. (11 mm) steel grinding ball (2156) per well. Case of 100 titer plates.

2230 24-Well Titer PlateSame as above, but a single titer plate.

2231-10 Cap-Mat for 2230 Titer Plate, Case of 10Cap-Mats seal 2230 24-Well Titer Plates. Case of 10 Cap-Mats.


Vials and Vial Sets

Vial sets are an alternative to titer plates. Vials can be filled and sealed individually, and the larger vials can hold bigger samples than titer plates. Replacement vials and steel balls can be purchased separately.

2240-PC Pre-Cleaned Polycarbonate Vial Set, 5 mL, Case of 10 Each set includes 24 pre-cleaned 2241-PC vials, each with a 3/8 in. (9.5 mm) steel grinding ball (2155). Ready to use, in a polyethylene carrier with foam vial holder. Carrier fits Geno/Grinder clamp; used in pairs. Grinding load per vial 1.5 mL. Vials can also be used with 2662 Cryo-Block or 2190 Vial Rack. Case of 10 sets, each set containing 24 vials pre-loaded with steel balls.

2240-PEF Pre-Cleaned Frosted Polyethylene Vial Set, 5 mL, Case of 10 Each set includes 24 vials with screw-cap. Ready to use, in a polyethylene carrier with foam vial holder. Carrier fits Geno/Grinder clamp; used in pairs. Grinding load per vial 1.5 mL. Holds one 3/8 in. (9.5 mm) steel grinding ball (2155). Vials can also be used with 2662 Cryo-Block or 2190 Vial Rack. Case of 10 sets, each set containing 24 vials.


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2241-PC Polycarbonate Vial, 5 mLPolycarbonate vial with screw-on polyethylene cap. ½ in. diameter x 2 in. long (12.7 mm x 50.8 mm). Holds one 3/8 in. (9.5 mm) steel grinding ball (2155), fits 2240-PC Vial Set, 2662 Cryo-Block, and 2190 Vial Rack. Not pre-cleaned or pre-loaded with grinding ball. Grinding load per vial 1.5 mL. Sold in units of 240.

2241-PEF-200 Frosted Polyethylene Vial, 5 Ml Frosted Polyethylene vial with screw-cap, 1/2 in. diameter x 2 in. long (12.7 mm x 50.8 mm). Holds one 3/8 in. (9.5 mm) steel grinding ball (2155). Can be used with 2662 Cryo-Block or 2190 Vial Rack. Not pre-cleaned or pre-loaded with grinding ball. Grinding load per vial 1.5 mL. Sold in units of 200.

2250 Pre-Cleaned Polycarbonate Vial Set, 15 mL Each set includes 5 pre-cleaned 2251PC vials, each with a 7/16 in. (11 mm) steel grinding ball (2156). Ready to use, in a polyethylene carrier with foam vial holder. Carrier fits Geno/Grinder clamp; used in pairs. Grinding load per vial 6 mL. Vials can also be used with 2660 Cryo-Block. Case of 10 sets, each set containing 5 vials pre-loaded with steel balls.

2251PC Polycarbonate Vial, 15 mL, ShortPolycarbonate vial with screw-on cap, 1 1/8 in. diameter x 1 2/3 in. long (2.9 cm x 4.2 cm). Holds all sizes of grinding media up to 7/16 in. (11 mm) steel grinding balls (2156). Fits 2250 Vial Set, 2660 Cryo-Block. Pre-cleaned. Grinding load per vial 6 mL. Sold in units of 100.

2252-PC-30 Polycarbonate Vial, 15 mL, Tall Polycarbonate vial with screw-on cap, 5/8 in. diameter x 4 ¾ in. long (1.6 cm x 12.1 cm). Holds grinding media up to 3/8 in. Fits 2197 Holder and 2661 Cryo-Block. For use with 2199 Large Clamp Assembly. Grinding load per vial 6 mL. Sold as a pack of 30 Vials.

2253-PC-48 Polycarbonate Vial, 50 mL Polycarbonate vial with screw-on cap, 1 1/8 in. diameter x 4 ½ in. (2.9 cm x 11.4 cm). Holds grinding media up to ½ in. Fits 2196-16-PE Holder and 2664 Cryo-Block. Grinding load per vial 20 mL. Sold as a pack of 48 Vials.

2253C-48 Cap for 2253-PC-48 Extra caps for the 50 mL Polycarbonate Vial. Sold as a pack of 48.

2254-PE-48 LDPE Vial, 50 mL Low density polyethylene vial with screw-on cap, 1 1/8 in. diameter x 4 ½ in. (2.9 cm x 11.4 cm). Holds grinding media up to ½ in. Fits 2196-16-PE Holder and 2664 Cryo-Block. Grinding load per vial 20 mL. Sold as a pack of 48 Vials.

2255 Cryogenic Grinding Vial Pack Used to grind tough materials (e.g. bone) at cryogenic temperatures. Package includes four sets of cryogenic vials, each with 1 cylinder, 2 end plugs, and 2 tungsten carbide balls. For use with 2199 Large Capacity Clamp Assembly and 2260 Cryo-Block. 6754 extractor required for end plug removal.

2255C4 Polycarbonate Center Cylinder, pack of 4Pack of 4 replacement cylinders for 2255 Grinding Vial Set.

2255C20 Polycarbonate Center Cylinder, pack of 20Pack of 20 replacement cylinders for 2255 Grinding Vial Set.

2301-100MB 2 mL Tube with 100 µm Molecular Biology Grade Silica Beads Self-standing 2 mL microfuge tube with screw-cap, prefilled with 100 µm Molecular Biology Grade Silica Beads. Package of 100.

2301-400MB 2 mL Tube with 400 µm Molecular Biology Grade Silica BeadsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 400 µm Molecular Biology Grade Silica Beads. Package of 100.

2301-800MB 2 mL Tube with 800 µm Molecular Biology Grade Silica BeadsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 800 µm Molecular Biology Grade Silica Beads. Package of 100.

2302-100AW1 2 mL Tube with 100 µm Acid Washed Zirconium BeadsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 1275 mg of 100 µm Acid Washed Zirconium Beads. Package of 100.

2302-100AW2 2 mL Tube with 100 µm Acid Washed Zirconium BeadsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 600 mg of 100 µm Acid Washed Zirconium Beads. Package of 100.

2302-100LB 2 mL Tube with 100 µm Low Binding Zirconium BeadsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 100 µm Low Binding Zirconium Beads. Package of 100.


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2302-200AW 2 mL Tube with 200 µm Acid Washed Zirconium BeadsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 200 µm Acid Washed Zirconium Beads. Package of 100.

2302-1700AW 2 mL Tube with 1.7 mm Acid Washed Zirconium BeadsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 1.7 mm Acid Washed Zirconium Beads. Package of 50.

2302-3000AW 2 mL Tube with 3 mm Acid Washed Zirconium BeadsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 3 mm Acid Washed Zirconium Beads. Package of 50.



2303-MM1 2 mL Tube with Mixed Matrix 1Self-standing 2 mL microfuge tube with screw-cap, prefilled with 500 μm Garnet and a 6 mm Zirconium Grinding Satellite. Package of 100.

2303-MM2 2 mL Tube with Mixed Matrix 2Self-standing 2 mL microfuge tube with screw-cap, prefilled with 800 μm and 1.4 mm Acid Washed Zirconium Beads. Package of 100.

2303-MM3 2 mL Tube with Mixed Matrix 3Self-standing 2 mL microfuge tube with screw-cap, prefilled with 100 μm Silica Beads, 1.4 mm Zirconium Beads and a 4 mm silica bead, Acid Washed. Package of 100.

2304-100AW 2 mL Tube with 100 µm Acid Washed Silica BeadsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 100 µm Acid Washed Silica Beads. Package of 100.



2305-2800SS 2 mL Tube with 2.8 mm Stainless Steel BallsSelf-standing 2 mL microfuge tube with screw-cap, prefilled with 2.8 mm Stainless Steel Grinding Balls. Package of 50.

3116PC 5 mL Polycarbonate VialPolycarbonate vial with slip-on polyethylene cap, ½ in. diameter x 2 in. long (12.7 mm x 50.8 mm). Holds one 3/8 in. (9.5 mm) steel grinding ball (2155), fits 2190 Vial Rack. Grinding load per vial 1.5 mL. Sold in units of 100.

Racks and Holders

These Racks and Holders are designed to hold vials securely in place on the Geno/Grinder during processing. They are lightweight and can also be used on the benchtop and to transport samples within the laboratory.

Racks2190 Vial RackHolds 24 3116PC vials, 24 2241-PC vials, or 24 2241-PEF-200 vials. All three vials hold a single 3/8 in. (9.5 mm) steel grinding ball (2155) to grind unusually large or tough samples.

Holders2196-16-PE Holder for 50 mL Centrifuge TubesThick, rugged foam block holds 16 standard 50 mL centrifuge tubes, 115 mm length. For use with 2199 Large Capacity Clamp Assembly.

2197 Holder for 15 mL Centrifuge TubesThick, rugged foam block holds 24 standard 15 mL centrifuge tubes, 119 mm length. For use with 2199 Large Capacity Clamp Assembly.

2198-24-PE Holder for 6133PC VialThick, rugged foam block holds 24 vials, ¾” x 2”.

2300 Holder for 2 mL microfuge TubesThick, rugged foam block holds 48 microfuge Tubes.


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Kryo-Tech®

These accessories are used to chill samples to cryogenic temperature and to maintain temperature during grinding. Additional products for liquid nitrogen handling, such as protective gloves and portable Dewars, can be found in the Freezer/Mill section.

2600 Cryo-Station A holding station for samples which must be kept chilled to preserve RNA for extraction.

The insulated outer jacket of the Cryo-Station is chilled with liquid nitrogen and can be filled automatically or manually. Two titer plates can be placed in the Cryo-Station well to be filled and kept cold. The well can also hold a chilled cutting board to cut up sensitive samples for RNA analysis.

Cryo-Blocks

Cryo-Blocks are made of aluminum. They hold and chill titer plates and vials for grinding plant and animal tissue for nucleic acid and protein extractions. Cryo-blocks can be quickly chilled in the 2600 Cryo-Station or directly in liquid nitrogen, and will keep samples cold during grinding to preserve RNA and proteins from heat degradation. The 2650, 2660, 2662, and 2665 Cryo-Blocks have about the same footprint and height as deep-well titer plates, and fit the standard clamp. The 2260, 2661 and 2664 Cryo-Blocks fit the 2199 Large Capacity Clamp. Cryo-Blocks are sold in pairs because they are used two at a time, to balance the load in the clamp.

2260 Cryo-Block for 2255 Vial, set of 2Aluminum block holds eight 2255 vials. Set includes two Cryo-Blocks, allowing 16 samples to be run simultaneously. For use with 2199 Large Capacity Clamp Assembly.

2650 Cryo-Adapter for Titer PlatesExtruded aluminum insert for 2210 Titer Plate. When inserted in titer plate and chilled in 2600 Cryo-Station, 2650 Cryo-Adapter keeps samples in titer plate cold during grinding, preserving RNA and proteins. Sold as a pair.

2660 Cryo-Block for 15 mL Polycarbonate Vials Aluminum block holds six 2251PC vials, as used in 2250 Vial Set. Cryo-Blocks are chilled in 2600 Cryo-Station to preserve RNA and proteins during grinding. Sold in pairs.

2661 Cryo-Block for 15 mL Centrifuge TubesAluminum block holds fifteen 15 mL conical bottom centrifuge tubes. Cryo-blocks are chilled in 2600 Cryo-Station to preserve RNA and proteins during grinding. Use with 2195 Large Capacity Clamp Assembly. Sold as a pair.

2662 Cryo-Block for 4 mL Polycarbonate VialsAluminum block holds twenty-four 2241-PC vials, as used in 2240-PC Vial Set. Cryo-Blocks are chilled in 2600 Cryo-Station to preserve RNA and proteins during grinding. Sold as a pair.

2663 Cryo-Block for 4 mL Polyethylene VialsAluminum block holds twenty-four 2241-PE vials, as used in 2240-PE Vial Set. Cryo-Blocks are chilled in 2600 Cryo-Station to preserve RNA and proteins during grinding. Sold as a pair.

2664 Cryo-Block for 50 mL Centrifuge TubesAluminum block holds six standard conical-bottom 50 mL centrifuge tubes for cryogenic milling. Cryo-Blocks are chilled in 2600 Cryo-Station to preserve RNA and proteins during grinding. Use with 2195 Large Capacity Clamp Assembly. Sold as a pair.

2665 Cryo-Block for Micro-centrifuge or PCR TubesAluminum block holds 96 standard (0.6 mL) micro-centrifuge tubes. Cryo-Blocks are chilled in 2600 Cryo-Station to preserve RNA and proteins during grinding. Sold as a pair.

2666 Cryo-Block for 48 Micro-centrifuge or PCR TubesAluminum block holds 48 standard (1.5 - 2.0 mL) microcentrifuge tubes. Cryo-Blocks are chilled in 2600 Cryo-Station to preserve RNA and proteins during grinding. Sold as a pair.

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Order Online 24/7: www.spexsampleprep.com

SPEX SamplePrep 6970EFM Enclosed Freezer/Mill®

6970M-115 Freezer/Mill – Single Grinding Coil for 115V/60Hz operation

6970M-230 Freezer/Mill – Single Grinding Coil for 230V/50Hz operation, CE Approved

6970D-115 Freezer/Mill – Dual Grinding Coils for 115V/60Hz operation

6970D-230 Freezer/Mill – Dual Grinding Coils for 230V/50Hz operation, CE Approved

• Totally enclosed cryogenic impact grinder with Auto-Fill System. Eliminates liquid nitrogen (LN) exposure

• Available with single (6970M) or dual (6970D) grinding chambers for increased sample throughput

• Grind up to 8 samples using the standard Freezer/Mill Grinding Vials.

• Grinds samples in the 0.1 to 100 gram range

• Each Coil holds one Large Vial, one Mid-Size Vial, or up to four standard Small Vials

• Color touch screen controls for programming grinding cycles and time, impact rate, and pre-cooling and cooling times

• Grinding chambers in 6970D can be programmed and operated independently or simultaneously

• Stores 10 grinding programs

• Safety features include LN sensor and door interlock

• Liquid nitrogen required for operation

• 6970D and 6970M supplied with choice of 6870L Accessory Package for Large Vials or 6870S Accessory Package for Small Vials

Specifications:

Dimensions: 22 in. (56 cm) x 18¼ in. (46 cm) x 17 in. (43 cm)

Weight: 70 lbs. (31.7 kg) net (6970D)



SPEX SamplePrep 6870 Freezer/Mill®

6870-115 Freezer/Mill – For 115V/60Hz operation

6870-230 Freezer/Mill – For 230V/50Hz operation, CE Approved

• Large cryogenic impact grinder with self-contained liquid nitrogen tub and insulated case.

• Touch-screen controls for programming grinding cycles, times, impact rate, pre-cooling and cooling times.

• Stores 10 grinding programs.

• Grinds samples in the 0.1 to 100 gram range.

• Coil holds one Large Vial, one Mid-Size Vial, or up to four Standard Small Vials.

• 6820 Auto-Fill System available for automated filling with liquid nitrogen.

• Safety features include LN sensor and lid interlock.

• Liquid nitrogen required for operation.

• 6870 Freezer/Mill supplied with choice of 6870L Accessory Package for Large Vials or 6870S Accessory Package for Small Vials.

• Accessory package for Mid-Sized Vials sold separately.

Specifications:

Dimensions: 20 ½ in. (52 cm) x 21 ½ in. (55 cm) x 18 in. (46 cm)

Weight: 42 lbs. (19 kg) net; 48 lbs. (22 kg) gross



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SPEX SamplePrep 6770 Freezer/Mill®

6770-115 Freezer/Mill – For 115V/60Hz operation

6770-230 Freezer/Mill – For 230V/50Hz operation, CE Approved

• Cryogenic impact grinder with self-contained liquid nitrogen tub and insulated case.

• Touch-screen controls for programming grinding cycles and time, impact rate, pre-cooling and cooling times.

• Stores 10 grinding programs.

• Grinds samples in the 0.1 gram to 5.0 gram range.

• Coil holds one standard Small Grinding Vial or one 6753 Microvial Set

• Pre-cooling chamber holds up to three additional vials.

• Safety features include LN sensor and lid interlock.

• Liquid nitrogen required for operation.

• 6770 Freezer/Mill supplied with 6754 Extractor/Vial Opener and 6755 Small Vial Rack.

Specifications:

Dimensions: 17 in. (43 cm) x 10½ in. (27 cm) x 13½in. (35 cm)

Weight: 19 lbs. (8.6 kg) net; 25 lbs. (11.3 kg) gross



Accessories

Vials & Vial SetsVials for the Freezer/Mill product line consist of a center cylinder, two end plugs, and an impactor. The center cylinder forms the grinding chamber, while the end plugs are removable for extraction of sample and cleaning. The solid, cylindrical impactor is magnetically shuttled from end to end in the vial, with grinding occurring as the sample is struck by the impactor against the end plug. Sample capacity is approximate. Actual sample capacity range for a vial depends on the nature of the material.

6751 Small Grinding Vial Set Set includes stainless steel impactor, two end plugs, and four polycarbonate center cylinders (6751C4). Set includes one complete vial plus three spare center cylinders. Transparent plastic allows visual check of grinding progress. Sample capacity 0.5 - 4.0 mL (0.1 - 5.0 g).

6751C4 Small Polycarbonate Center CylinderFor 6751 vial set; sold in package of 4. Polycarbonate is durable but should be cleaned with hot water and detergent, not organic solvents. It can be sterilized with a dilute bleach solution.

6751C20 Small Polycarbonate Center CylinderSame as above, but a package of 20.

6752 Small Steel Center Cylinder For 6751 vial set. Replaces 6751C when sample contact with plastic is not advised. Made of nonmagnetic stainless steel.

6753 Microvial Set Three 6753V Microvials with holder. Each 6753V has stainless steel end plugs, center cylinder and impactor. Holder contains three 6753V Microvials for simultaneous operation. Sample capacity 0.1 - 0.5 mL (0.1 - 0.5 g) each.

6761 Small Poly-Vial Set For grinding soft tissues without metal contamination. End plugs and center cylinder are polycarbonate, impactor is polycarbonate-encapsulated steel. Set includes one complete vial plus three spare center cylinders. Sample capacity 0.5 - 4.0 mL (0.1 - 5.0 g).

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6771 Small Cr-Free Vial Set For RoHS/WEEE testing of electronic components for chromium. End plugs and impactor are Cr-free ASTM 06 steel. Will not add Cr to sample like stainless steel components of 6751 Vial. Set includes three extra 6751C cylinders. Sample capacity 0.5 - 4.0 mL (0.1 - 0.5 g).

6781S Small Stainless Steel Grinding Vial SetSet includes stainless steel impactor, stainless steel center cylinder, two end plugs, and pack of 100 O-rings. O-rings are used on end plugs to prevent sample leakage during grinding. For use when contact with plastic is not recommended. Made of non-magnetic stainless steel.

6801 Large Grinding Vial Set Set includes stainless steel impactor and two end plugs, plus four polycarbonate center cylinders (6801C4). Set includes complete vial plus three spare center sections. Transparent plastic allows visual check of grinding progress. Sample capacity up to 50 mL (1 - 100 g).

6801C4 Large Polycarbonate Center Cylinder For 6801 and 6871 Vial Sets. Polycarbonate is durable but should be cleaned with detergent and water, not organic solvents. It can be sterilized with a dilute bleach solution. Sold in units of 4.

6801C20 Large Polycarbonate Center CylinderSame as 6801C4, but a package of 20.

6802 Large Steel Center Cylinder For 6801 Vial Set. Used in place of 6801C when polycarbonate cylinder is not compatible with sample. Made of nonmagnetic stainless steel.

6871 Large Cr-Free Vial SetFor grinding electronic components to test for chromium under RoHS/WEEE directives. Cr-free ASTM 06 steel impactor and end plugs, polycarbonate center cylinder (6801C). Will not add Cr like stainless steel components of 6801 vial. Set includes one complete vial plus three extra cylinders. Sample capacity up to 50 mL (1 - 100 g).

6881 Mid-Size Vial Set Grinding vial intermediate in size between 6751 and 6801: sample capacity up to 25 mL (1 - 40 g). Stainless steel end plugs and impactor, polycarbonate center cylinder (6881C). Set includes one complete vial plus three extra cylinders. Must be used with 6884 and 6886 Adapters.

6881C4 Mid-Size Polycarbonate Center CylinderFor 6881, 6883, and 6885 Vial Sets. Polycarbonate is durable but should be cleaned with detergent and water, not organic solvents. Sold in units of 4.

6881C20 Mid-Size Polycarbonate Center CylinderSame as 6881C4, but a package of 20.

6883 Mid-Size Cr-Free VialFor grinding electronic components to test for chromium under RoHS/WEEE directives. Cr-free ASTM 06 steel impactor and end plugs, polycarbonate center cylinder (6881C). Vial set includes one complete vial and three spare cylinders. Sample capacity up to 25 mL (1 - 40 g). Must be used with 6884 and 6886 Adapters.

6885 Mid-Size Poly-VialFor grinding soft tissues without metal contamination. End plugs and center cylinder are polycarbonate, impactor is polycarbonate-encapsulated steel. Vial set includes one complete vial plus three extra center cylinders. Sample capacity up to 25 mL (1 - 40 g). Must be used with 6884 and 6886 Adapters.

Vial Racks6755 Vial Rack for Small VialsFor 6751, 6753, 6761, and 6771 Vial Sets. Glass-reinforced acetal rack holds up to sixteen vials for storage and handling. One 6755 Vial Rack is supplied with each 6770 Freezer/Mill and 6870S Accessory Package.

6805 Vial Rack for Large VialsEpoxy-coated steel rack holds six 6801 or 6871 Vial Sets for storage and handling. One 6805 is supplied with each 6870L Accessory Package.

Extractors6814 AutoExtractor™ for Freezer/Mill® Vial Sets

6814-115 AutoExtractor™ – For 115 V/60 Hz operation.

6814-230 AutoExtractor™ – For 230 V/50 Hz operation.The 6814 AutoExtractor is an electrically powered version of our 6754 and 6804 Extractor/Vial Openers. Cold vials can now be opened easily. The 6814 will open large Vial Sets (6801, 6871), and includes a special adapter for small Vial Sets (6751, 6753, 6761, 6771). Increases throughput, reduces operator effort. Not adaptable for Mid-Size Vial Sets.

6753C Microvial Sample Extraction Tool For removing ground samples from Microvials. The 6753C is inserted into an individual vial, then pushed through to clear sample from the tube. Shown with 6753V Microvial.

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6754 Extractor/Vial Opener for Small Vials The Extractor/Vial Opener lifts 6700-series vials in and out of the Freezer/Mill, and is used to remove the vial end plugs from the center section. One 6754 is supplied with each 6770 Freezer/Mill and as part of the 6870S Accessory Package.

6804 Extractor/Vial Opener for Large and Mid-Sized Vials The Extractor/Vial Opener lifts vial sets in and out of the Freezer/Mill, and is used to remove the vial end plugs from the center section. Lever assists end-plug extraction. One

6804 is supplied with each 6870L Accessory Package. Add 6884 Adapter for use with Mid-Size Vials.

Sample Adapters6806 Multi-Vial Adapter Required when using one to four 6751, 6761, or 6771 Vials in the 6870 or 6970EFM Freezer/Mills. The Multi-Vial Adapter is an insert placed inside the coil to separate and align small vials during grinding. One 6806 is supplied with each 6870S Accessory Package.

6884 Mid-Size Vial Adapter for 6804 Extractor/Vial Opener Adapts 6804 Extractor/Vial Opener for use with Mid-Size Vials. Insert attaches to bell of 6804, is easily installed, removed.

6886 Mid-Size Vial Adapter for 6800-Series Freezer/MillsPositions and aligns Mid-Size vials in 6800, 6850, 6870, and 6970EFM Freezer/Mills.

Accessory Packages & KitsEach 6970EFM or 6870 Freezer/Mill is supplied with your choice of 6870L Large Vial Accessory Package or 6870S Small Vial Accessory Package. The 6870 Freezer/Mill can hold up to four small vials* (6751, 6761, 6771) or one mid-size vial (6881, 6883, 6885). The Accessory Package for mid-size vials must be purchased separately. Mid-size vials require the 6870M Mid-Size Vial Accessory Package, consisting of one 6884 Mid-Size Vial Adapter for 6804 Extractor/Vial Opener, and one 6886 Mid-Size Vial Adapter for 6800-Series or 6970EFM Freezer/Mills. Small vials require the 6870S Small Vial Accessory Package, consisting of one 6754 Extractor/Vial Opener, one 6755 Small Vial Rack, and one 6806 Multi-Vial Adapter.

*Note: the 6753 Microvial Set can only be run one at a time in the 6870 Freezer/Mill.

6870S Small Vial Accessory Package For 6751, 6753, 6761, and 6771 Vial Sets. Includes 6754 Extractor/Vial Opener, 6755 Vial Rack, and 6806 Multi-Vial Adapter.

6870M Mid-Size Vial Accessory Package For 6881, 6883, and 6885 Mid-Size Vial Sets. Includes 6884 Mid-Size Vial Adapter for 6804 Extractor/Vial Opener, and 6886 Mid-Size Vial Adapter for large Freezer/Mills.

6870L Large Vial Accessory Package For 6801 and 6871 Vial Sets. Includes 6804 Extractor/Vial Opener and 6805 Vial Rack.

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Liquid Nitrogen HandlingOur products are designed to aid the user in the safe handling of liquid nitrogen. The boiling point of liquid nitrogen is -195.80º C. When working with liquid nitrogen directly or indirectly, cryogenic gloves must be worn to protect hands. Our 6820 Auto-Fill System is an option for filling the 6870 Freezer/Mill without direct LN contact. We also offer transfer hoses as well as 25 L and 50 L Dewars for dispensing LN.

6820 Auto-Fill System for the 6870 Freezer/MillThe 6820 Auto-Fill System is designed to automatically maintain the required level of liquid nitrogen within the 6870 Freezer/Mill during operation. The Auto-Fill System is a more efficient and safer way to handle the volume of liquid nitrogen required for processing samples in the 6870 Freezer/Mill. This automated system reduces the loss of liquid nitrogen and saves the time lost when manually filling the reservoir.

The Auto-Fill System requires a cryogenic transfer hose, such as the SPEX SamplePrep 6906 or 6907 Cryogenic Transfer Hoses, which are sold separately. Transfer hoses are available in different lengths and can be constructed with different materials. The transfer hose must be configured with a ½ inch (12.7 mm) JIC female flare nut at each end for proper installation. The 6820 package does include an adapter to allow compatibility with the 6906 and 6907 Cryogenic Transfer Hoses.

The Auto-Fill System is also supplied with a safety relief valve that connects to the liquid nitrogen supply via a ½ inch (12.7 mm) JIC female flare nut. The safety relief valve attaches at the high point of the transfer hose, and vents nitrogen gas pressure that may build up in the hose when there is liquid nitrogen in the hose and the valves at both ends are closed.

The 6820 Auto-Fill System should be ordered at the same time as the 6870 Freezer/Mill so it can be installed and calibrated at our factory.

6900 Cryogenic Gloves Designed to protect the hands and arms from the hazards associated with ultra-cold materials. Highly recommended for safe operation of SPEX SamplePrep Freezer/Mills. These multi-layer-insulated gloves are lightweight and flexible, allow excellent dexterity, and are comfortable to wear for extended periods of time. The gloves are sold as a pair and are available in small (6900S), medium (6900M), large (6900L) and X-large (6900XL).

6905 Portable Cryogenic Dewar, 25 L Capacity Cryogenic Dewar designed for storing and dispensing small amounts of liquid nitrogen. Sturdy, aluminum, five-wheel roller base makes moving Dewar from room to room safe and easy. Includes liquid withdrawal device for dispensing liquid nitrogen directly into the Freezer/Mill at rates up to 8 liters per minute. Capacity 25 liters. Compatible with 6806 and 6807 Cryogenic Transfer Hoses.

6910 Portable Pressurized Cryogenic Cylinder, 50 L CapacityCryogenic cylinder designed for storing and dispensing liquid nitrogen with pressure of up to 22 psi. Sturdy, stainless steel, four-wheel design makes moving the cylinder from room to room, safe and easy. Perfect for use with the 6970EFM or 6870 with autofill. Capacity 50 liters. Compatible with 6906 and 6907 transfer hoses.

6906 Short Cryogenic Transfer Hose4 ft. (1.2 m) long flexible stainless steel hose suitable for transferring cryogenic fluids such as liquid nitrogen. Fitted with a 3/8 in. (9.5 mm) NPT male fitting on one end and a CGA295 female fitting on the other end. Compatible with 6905 and 6910 Portable Cryogenic Dewar, 6970EFM and 6820 Auto-Fill system.

6907 Long Cryogenic Transfer Hose6 ft. (1.8 m) long flexible stainless steel hose suitable for transferring cryogenic fluids such as liquid nitrogen. Fitted with a 3/8 in. (9.5 mm) NPT male fitting on one end and a CGA295 female fitting on the other end. Compatible with 6905 and 6910 Portable Cryogenic Dewar, 6970EFM and 6820 Auto-Fill system.

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SPEX SamplePrep 8000D Dual Mixer/Mill®

8000D-115 Mixer/Mill - For 115 V/60 Hz operation

8000D-230 Mixer/Mill - For 230 V/50 Hz operation, CE Approved

• Efficient two-clamp laboratory mill for pulverizing hard, brittle samples

• Also called a shaker mill or high-energy ball mill

• Sample sizes in the 10 gram range for grinding, and up to 60 mL for blending powders or mixing emulsions

• Reduces samples to analytical fineness

• Can be utilized for mechanical alloying and nanomilling

• Features include a variable-range electronic timer, dual clamps for multiple sample capacity, safety interlock system, and forced-air cooling

• Timer is factory-set for 100-minute range. Optional chip is available to extend timer range to 1,000 or 10,000 minutes

• For efficiency, clamps are run in balance with the same vial and load in each clamp

• Accepts grinding/mixing vials from 2 3/8 in. (6 cm) to 3 1/8 in. (7.9 cm) in length and up to 2 ¼ in. (5.7 cm) in diameter

• The 8010 and 8011 adapters permit processing of multiple samples in ½ in. x 1 in. (12.7 mm x 25.4 mm), ½ in. x 2 in. (12.7 mm x 50.8 mm), and ¾ in. x 2 in. (19.1 mm x 50.8 mm) vials

Specifications:

Dimensions: 24 in. (61 cm) x 17 in. (43 cm) x 11 in. (28 cm)

Net Weight: 82 lb. (37 kg)

Gross Weight: 98 lb. (44 kg)

Voltage: 115 V/60 Hz or 230 V/50 Hz

Motor: 1/3 HP, 1725 RPM @ 60 Hz or 1425 RPM @ 50 Hz

Controls: Start and stop buttons, programmable 100-minute timer with digital display.

Power Cord: 3-prong grounded plug for 115 V/60 Hz version 2-prong European plug for 230 V/50 Hz version

Clamp Movement: 2 1/3 in. (5.9 cm) back-and-forth, 1 in. ( 2.5 cm) side-to-side Clamp speed in complete back-and-forth cycles per minute: 1060 (115 V/60 Hz) or 875 (230 V/50 Hz)

SPEX SamplePrep 8000M Mixer/Mill®

8000M-115 Mixer/Mill - For 115 V/60 Hz operation

8000M-230 Mixer/Mill - For 230 V/50 Hz operation, CE Approved

• Also called a shaker mill or high-energy ball mill

• Sample sizes in the 10-gram range for grinding, and up to 60 mL for blending powders or mixing emulsions

• Reduces samples to analytical fineness

• Can be utilized for mechanical alloying and nanomilling

• Features include a variable-range electronic timer, safety interlock system, and shock-mounted electric motor for tabletop operation

• Timer is factory-set for 100-minute range. Optional chip is available to extend timer range to 1,000 or 10,000 minutes

• Accepts grinding/mixing vials from 1 ¾ in. (4.5 cm) to 3 1/8 in. (7.9 cm) in length and up to 2 ¼ in. (5.7 cm) in diameter

• The 8010 and 8011 adapters permit processing of multiple samples in ½ in. x 1 in. (12.7 mm x 25.4 mm), ½ in. x 2 in. (12.7 mm x 50.8 mm), and ¾ in. x 2 in.(19.1 mm x 50.8 mm) vials

Specifications:


Weight: 76 lbs. net (35 kg); 92 lbs. gross (41 kg)



Controls: Start and stop buttons, programmable 100-minute timer with digital display.

Power Cord: 3-prong standard North American plug for 115 V/60 Hz version 2-prong European CEE 7/7 plug for 230 V/50 Hz version

Clamp Movement: 2 1/3 in. (5.9 cm) back-and-forth, 1 in. (2.5 cm) side-to-side Clamp speed in complete back-and-forth cycles per minute: 1060 (115 V/60 Hz) or 875 (230 V/50 Hz)

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SPEX SamplePrep 5100 Mixer/Mill®

5100-115 Mixer/Mill – For 115 V/60 Hz operation

5100-230 Mixer/Mill – For 230 V/50 Hz operation• High-energy “shaker mill”

• Pulverizes and/or homogenizes samples of varying composition

• Grinding containers constructed with a variety of materials

• Typically grinds samples in the 0.5 gram to 1.5 gram range

• Holds three vials of ½ in. (12.7 mm) diameter or two vials of ¾ in. (19.1 mm) diameter (length of vials up to 2¼ in. (57.2 mm).

• For grinding up to 3 mL or blending up to 10 mL of sample

• Push-button, resettable 30-minute timer on the 5100-115 Mixer/Mill

• Push-button, resettable 72-minute timer on the 5100-230 Mixer/Mill

Specifications:


Weight: 15.2 lbs. net (7 kg), 19.8 lbs. gross (9 kg)



Controls: Push-button, resettable 30-minute timer on 115 V/60 Hz version Push-button, resettable 72-minute timer on 230 V/50 Hz version


Clamp Movement: ½ in. (12.7 mm) front-to-back, 3/8 in. (9.5 mm) side-to-side Clamp speed in complete back-and-forth cycles per minute: 3000 (115 V/60 Hz or 2500 (230 V/50 Hz)

Mixer Mill Accessories

Vials & Vial SetsMixer/Mill vials are available individually or as part of a set which includes end caps and grinding media. They are available in a wide variety of sizes and materials for maximum on-the-job flexibility over a wide range of applications.

Small Vials and Vial SetsFor use in all Mixer Mills (8000 Series requires adapter)

3111 Polystyrene Vial ½ in. (12.7 mm) diameter x 1 in. (25.4 mm) long. Includes slip-on polyethylene cap. Volume 2.5 mL; mixing load approximately 1 mL. Accepts SPEX SamplePrep 3112 or 3119 methacrylate balls. Sold in units of 100.

3116 Polystyrene Vial ½ in. (12.7 mm) diameter x 2 in. (50.8 mm) long. Includes slip-on polyethylene cap. Volume 5 mL; mixing load approximately 2 mL. Accepts SPEX SamplePrep 3112 or 3119 methacrylate balls. Sold in units of 100.

3116PC Polycarbonate Vial½ in. (12.7 mm) diameter x 2 in. (50.8 mm) long. Includes slip-on polyethylene cap. Volume 5 mL; mixing load approximately 2 mL. Accepts SPEX SamplePrep 3112 or 3119 methacrylate balls. Sold in units of 100.

6133 Polystyrene Vial ¾ in. (19.1 mm) diameter x 2 in. (50.8 mm) long. Includes slip-on polyethylene cap. Volume 12 mL; mixing load approximately 5 mL. Accepts SPEX SamplePrep 3112, 3119, or 8006A methacrylate balls. Sold in units of 100.

6133PC Polycarbonate Vial¾ in. (19.1 mm) diameter x 2 in. (50.8 mm) long. Includes slip-on polyethylene cap. Volume 12 mL; mixing load approximately 5 mL. Accepts SPEX SamplePrep 3112, 3119, and 8006A methacrylate balls. Sold in units of 100.

3114 Stainless Steel Vial Set ½ in. (12.7 mm) diameter x 1 in. (25.4 mm) long. Made of 303 stainless steel; includes slip-on cap and ¼ in. (6.35 mm) stainless steel ball. Volume 2.5 mL; grinding load 0.2–0.6 mL; mixing load 1 mL.

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3117 Hardened Tool Steel Vial Set ½ in. (12.7 mm) diameter x 1 in. (25.4 mm) long. Made of hardened tool steel. Includes slip-on cap and ¼ in. (6.35 mm) steel ball. Volume 2.5 mL; grinding load 0.2 – 0.6 mL; mixing load 1 mL.

3127 Hardened Tool Steel Vial Set ¾ in. (19.1 mm) diameter x 1 7/8 in. (47.6 mm) long. Made of hardened tool steel. Includes center cylinder with two slip-on caps and 1/4 in. (6.35 mm) steel ball. Volume 5 mL; grinding load 0.5–1.0 mL; mixing load 2 mL.

3118 Agate Vial Set 5/8 in. (15.9 mm) diameter x 1 1/3 in. (33 mm) long. Handmade from flawless Brazilian agate; must be run inside cushioned SPEX SamplePrep 6133 vial (included); includes stopper and ¼ in. (6.35 mm) agate ball. Volume 1.6 mL; grinding load 0.2–0.6 mL; mixing load 1 mL.

5004 Tungsten Carbide-Lined Vial Set ¾ in. (19.1 mm) diameter x 2 1/8 in. (54 mm) long. Includes two slip-on Delrin® caps with tungsten carbide inserts, six disposable methacrylate center cylinders, and two 5/16 in. (7.9 mm) tungsten carbide balls. Volume 5 mL; grinding load 0.5–1.5 mL; mixing load 3 mL.

5004C Methacrylate Center CylinderDisposable methacrylate center cylinder; for SPEX SamplePrep 5004 Vial Set. Sold in units of 20.

Large Vials and Vial SetsFor use in 8000 Series Mixer/Mills

8001 Hardened Steel Vial Set For wet or dry grinding/mixing. Vial size 2 ¼ in. (5.7 cm) diameter x 3 in. (7.62 cm) long. Vial body and cap liner made of hardened tool steel. Set includes screw-on cap with O-ring, two ½ in. (12.7 mm) and four ¼ in. (6.35 mm) steel balls. Volume 65 mL; grinding load 3–10 mL; mixing load approximately 25 mL.

8002 Polystyrene Vial For wet or dry mixing of liquids and powders. Vial size 2 1/8 in. (5.4 cm) diameter x 2 ½ in. (6.7 cm) long. Polystyrene body with plastic screw-on cap. Volume 135 mL; mixing load approximately 50 mL. For use with SPEX SamplePrep 3112 or 8006A methacrylate balls. Sold in units of 100.

8003 Alumina Ceramic Vial Set For wet (use 8015 Clamp) or dry grinding/mixing. Vial size 2 ¼ in. (5.7 cm) diameter x 2 ¾ in. (7.0 cm) long. Set includes high purity 99.5% alumina ceramic vial body and two slip-on caps, one ½ in. (12.7 mm) alumina ceramic ball, and eight corprene gaskets. Volume 45 mL; grinding load 2–8 mL; mixing load approximately 20 mL.

8015 Clamp for 8003 Vial SetClamps permit slurry grinding in SPEX SamplePrep 8003 Vial Set; sold in pairs. One pair required for each 8003.

8004 Tungsten Carbide Vial Set For wet or dry grinding/mixing. Vial size 2 ¼ in. (5.7 cm) diameter x 2 ½ in. (6.35 cm) long. Set includes tungsten carbide-lined body, two screw-on tungsten carbide-lined caps, two 7/16 in. (11.2 mm) tungsten carbide balls, and eight corprene gaskets. Volume 55 mL; grinding load 3–10 mL; mixing load approximately 25 mL.

8004SS Steel-Jacketed Tungsten Carbide Vial Set For wet or dry grinding/mixing in extreme environments. Vial size 2 ¼ in. (5.7 cm) diameter x 2 ½ in. (6.35 cm) long. The screw-on caps and body are 304 stainless steel, lined with tungsten carbide. Gaskets are Viton for enhanced resistance to heat and chemicals. Set includes two 7/16 in. (11.2 mm) tungsten carbide balls and six extra Viton gaskets. Volume 55 mL; grinding load 3-10 mL; mixing load approximately 25 mL.

8005 Zirconia Ceramic Vial Set For wet or dry grinding/mixing. Vial size 2 ½ in. (6.35 cm) diameter x 2 11/16 in. (6.8 cm) long. Set includes solid zirconia ceramic vial and slip-on cap, two ½ in. (12.7 mm) zirconia ceramic balls, and seven corprene gaskets. Volume 45 mL; grinding load 2–8 mL; mixing load approximately 20 mL.

8006 Methacrylate Vial Set For wet or dry mixing of liquids and powders. Vial size 2 ¼ in. (5.7 cm) diameter x 2 5/8 in. (6.7 cm) long. Rugged construction allows grinding of soft materials without metallic contamination. Heavy-gauge methacrylate body with methacrylate-lined aluminum screw-on cap. Set includes corprene gasket, two 8006A ½ in. (12.7 mm) and four 3112 3/8 in. (9.5 mm) methacrylate balls. Volume 55 mL; mixing load approximately 25 mL.

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8007 Stainless Steel Vial Set For wet or dry grinding/mixing. Vial size 2 ¼ in. (5.7 cm) diameter x 3 in. (7.62 cm) long. Vial body and cap liner made of hardened 440C stainless steel. Set includes screw-on cap with O-ring, two ½ in. (12.7 mm) and four ¼ in. (6.35 mm) stainless steel balls. Volume 65 mL; grinding load 3–10 mL; mixing load approximately 25 mL.

8008 Silicon Nitride Vial Set For wet or dry grinding/mixing. Vial size 2 ½ in. (6.35 cm) diameter x 2 11/16 in. (6.8 cm) long. Set includes solid silicon nitride vial and slip-on cap, two ½ in. (12.7 mm) silicon nitride balls, and seven corprene gaskets. Volume 45 mL; grinding load 2–8 mL; mixing load approximately 20 mL.

8009 Round-Ended Hardened Steel Vial Set For wet or dry grinding/mixing. Vial size 2 3/8 in. (6.0 cm) diameter x 3 in. (7.62 cm) long. Hardened steel vial body has grinding chamber with rounded ends for more efficient grinding/mixing. Includes screw-on cap, O-ring, two ½ in. (12.7 mm) and four ¼ in. (6.35 mm) steel balls. Volume 35 mL; grinding load 3–10 mL; mixing load approximately 25 mL.

8014 Agate Vial Set For dry grinding/mixing. Vial size 2 ¼ in. (5.7 cm) diameter x 2 ¾ in. (7.0 cm) long. Set includes all-agate vial body, two slip-on caps, two ½ in. (12.7 mm) agate balls, and eight corprene gaskets. Volume 45 mL; grinding load 3–8 mL; mixing load approximately 20 mL.

6134 Polystyrene Vial For wet or dry mixing. Vial size 1 in. (2.54 cm) diameter x 3 in. (7.62 cm) long. Includes polyethylene slip-on cap. Volume 35 mL; mixing load approximately 15 mL. For use with SPEX SamplePrep 3112 or 8006A methacrylate balls. Sold in units of 100.

6135 Polyethylene Vial For wet or dry mixing. Vial size 1 ½ in. (3.81 cm) diameter x 3 in. (7.62 cm) long. Polyethylene vial with attached cap. Volume 75 mL; mixing load approximately 40 mL. For use with SPEX SamplePrep 3112 or 8006A methacrylate balls. Sold in units of 100

Comparison of Grinding Vials

Grinding Typical Grinding Blending Sample Suitable for Slurry P/N Sample Volume Sample Weight Volume Grinding or Blending

Hardened Steel

3117 0.2-0.6 mL 0.5 g 1 mL No3127 0.5-1.0 mL 1 g 2 mL No8001 3-10 mL 10 g 25 mL Yes8009 3-10 mL 10 g 25 mL Yes

Stainless Steel

3114 0.2-0.6 mL 0.5 g 1 mL No8007 3-10 mL 10 g 25 mL Yes

Tungsten Carbide

5004 0.5-1.5 mL 1 g 2.5 mL Yes8004 3-10 mL 10 g 25 mL Yes8004SS 3-10 mL 10 g 25 mL Yes

Alumina Ceramic

8003 2-8 mL 10 g 20 mL YesAgate

3118 0.2-0.6 mL 0.5 g 1 mL No8014 5-10 mL 10 g 25 mL No

Zirconia Ceramic

8005 2-8 mL 10 g 20 mL YesSilicon Nitride

8008 2-8 mL 10 g 20 mL YesPlastic

3111 - - 1 mL Yes3116 - - 2 mL Yes3116PC - - 2 mL Yes6133 - - 5 mL Yes6133PC - - 5 mL Yes6134 - - 15 mL Yes6135 - - 40 mL Yes8002 20-50 mL - 50 mL Yes8006 3 – 10 mL - 20 mL Yes

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Grinding Media3112 Methacrylate Balls Methacrylate Balls 3/8 in. (9.5 mm) diameter; fit all SPEX SamplePrep plastic vials. Sold in units of 100.

3119 Methacrylate Balls Methacrylate Balls 1/8 in. (3.2 mm) diameter; fit all SPEX SamplePrep plastic vials. Sold in units of 100.

8006A Methacrylate Balls Methacrylate balls ½ in. (12.7 mm) diameter; fit all SPEX SamplePrep plastic vials ¾ in. (19.1 mm) diameter and larger. Sold in units of 100.

3114SB Stainless Steel Ball Set ¼ in. (6.35 mm) diameter stainless steel (440C) ball; for SPEX SamplePrep 3114 Vial Set. Four balls per set.

3117B Hardened Tool Steel Ball Set¼ in. (6.35 mm) inch diameter steel ball; for SPEX SamplePrep 3117 and 3127 Vial Sets. Four balls per set.

3118A Agate Ball¼ in. (6.35 mm) diameter agate ball; for SPEX SamplePrep 3118 Vial Set.

5004A Tungsten Carbide Ball Set5/16 in. (7.9 mm) diameter tungsten carbide ball; for SPEX SamplePrep 5004 Vial Set. Four balls per set.

8001B Steel Ball SetTwo ½ in. (12.7 mm) and four ¼ in. (6.35 mm) hardened steel balls for SPEX SamplePrep 8001 and 8009 Vial Sets.

8003A Alumina Ceramic Ball½ in. (12.7 mm) diameter alumina ceramic ball for SPEX SamplePrep 8003 Vial Set.

8004A Tungsten Carbide Ball7/16 in. (11.2 mm) diameter tungsten carbide ball for SPEX SamplePrep 8004 and 8004SS Vial Sets.

8005A Zirconia Ceramic Ball½ in. (12.7 mm) diameter zirconia ceramic ball for SPEX SamplePrep 8005 Vial Set.

8007B Stainless Steel Ball SetTwo ½ in. (12.7 mm) and four ¼ in. (6.35 mm) hardened 440C stainless steel balls for SPEX SamplePrep 8007 Vial Set.

8008A Silicon Nitride Ball½ in. (12.7 mm) diameter silicon nitride ball for SPEX SamplePrep 8008 Vial Set.

8014A Agate Ball½ in. (12.7 mm) diameter agate ball for SPEX SamplePrep 8014 Vial Set.

Sample AdaptersFor 8000 Series Mixer/MillsSpecial adapters enable simultaneous running of multiple samples in the 8000 Series Mixer/Mills. The 8010 and 8011 multiple-sample adapters accommodate ½ in. (12.7 mm) and 3/4 in. (19.1 mm) diameter vials, respectively.

8010 Multiple-Sample Adapter Holds seven vials, each ½ in. (12.7 mm) in diameter, for simultaneous running in 8000M Mixer/Mill. Suitable vials include SPEX SamplePrep 3111, 3114, 3116 and 3117. Vials run at one time should be same length. Can be used in 8000D Mixer/Mill with 3116 vials, or with 3111, 3114, and 3117 vials if 8012 adapter is also used.

8011 Multiple-Sample Adapter Holds four vials, each ¾ in. (19.1 mm) in diameter, for simultaneous running in 8000M Mixer/Mill. Suitable vials include SPEX SamplePrep 3118, 3127, 5004 and 6133. Vials run at one time should be same length. Can be used in 8000D Mixer/Mill with 3118, 5004, and 6133 vials. To use 3127 vials in 8000D, 8012 adapter is also necessary.

8012 Vial Clamp Adapter for the 8000D Mixer/Mill Required when running 8010 adapter with vials 1 in. (25.4 mm) in length (3111, 3114, 3117) or when running 3127 vials with 8011 adapter in the 8000D.

SPEX SamplePrep PrepAid® Grinding Aids3640 PrepAid Propylene Glycol – (C3H8O2) Liquid grinding aid, 25 grams (approximately 24 mL) in dropper bottle. Instructions included.

3650 PrepAid Vertrel® XF Liquid fluorocarbon grinding aid. Prevents caking, reduces contamination, evaporates after grinding. One-quart bottle. Instructions included.

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SPEX SamplePrep 8530 Enclosed Shatterbox®

8530-115 Enclosed Shatterbox® - For 115 V/60 Hz operation

8530-230 Enclosed Shatterbox® - For 230 V/50 Hz operation, CE Approved

• Features a cam-activated clamp that requires only moderate hand pressure to secure the grinding dish in the Shatterbox and is adjustable to secure containers of different heights

• LCD display with environmentally safe, push-button membrane switch

• Soundproof enclosure reduces noise levels

• Equipped with a safety interlock system and lockable casters

• Wide choice of grinding containers, sold separately

Specifications:

Dimensions: 19 in (48 cm) x 19 in (48 cm) x 40 in (102 cm)

Weight 225 lbs. gross (102 kg)


Motor: ½ horsepower @ 835 RPM


SPEX SamplePrep 8500 Shatterbox®

8500-115 Shatterbox® – For 115 V/60 Hz operation

8500-230 Shatterbox® – For 230 V/50 Hz operation• Features a cam-activated clamp that requires only moderate

hand pressure to secure the grinding dish in the Shatterbox and is adjustable for containers of different heights

• Perforated metal skirt provides ventilation for continuous operation, and a rubber cover protects against dust

• Includes an anti-skid wood/rubber floor mount and an extra set of drive belts

• Grinding containers are sold separately. Timer is not supplied

Specifications:

Dimensions: 13 in. (33 cm) x 13 in.(33 cm) x 23 1/2 in. (60 cm)

Weight 135 lbs. gross (61 kg)


Motor: ½ horsepower @ 835 RPM


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Accessories

Grinding Dishes8501 Hardened Steel Grinding Container Grinding container with puck, dish, and lid made of 52-100 hardened tool steel. 6.9 in. diameter x 2.9 in. high, 17.5 lbs. (17.5 cm x 7.5 cm, 8 kg). Nominal volume 340 mL, recommended grinding load 20-50 mL. Includes puck, ring, dish with O-ring gasket, and lid.

8504 Tungsten Carbide Grinding ContainerGrinding container lined with tungsten carbide. Dish and lid are steel shells with tungsten carbide inserts; puck and ring are solid tungsten carbide. 6.9 in. diameter x 3.2 in. high, 29 lbs. (17.5 cm x 8 cm, 12.5 kg). Nominal volume 240 mL, recommended grinding load 20-50 mL. Includes puck, ring, dish with O-ring gasket, and lid.

8505 Alumina Ceramic Grinding Container Grinding container lined with 99.5% alumina ceramic. Steel lid and aluminum dish are lined with alumina ceramic plates, puck is solid alumina ceramic. 6.3 in. diameter x 3.2 in. high, 11 lbs. (15.2 cm x 8 cm, 5 kg). Nominal volume 170 mL, recommended grinding load 15-40 mL. Includes puck, dish with O-ring gasket, and lid.

8506 Zirconia Ceramic Grinding Container Grinding container lined with Mg-stabilized zirconia ceramic. Steel lid and aluminum dish are lined with zirconia ceramic, puck is solid zirconia ceramic. 6 in. diameter x 3.2 in. high, 13 lbs. (15.2 cm x 8 cm, 6 kg). Nominal volume 170 mL, recommended grinding load 15-40 mL. Includes puck, dish with O-ring gasket, and lid.

8507 Small Hardened Steel Grinding ContainerSmall grinding container with puck, dish and lid made of 52-100 hardened steel, 3.6 in. diameter x 2.3 in high, 4 lbs. (9 cm x 5.4 cm, 1.9 kg). Nominal volume 90 mL, recommended grinding load 2-20 mL. Includes puck, dish with gasket, and lid. Must be used either one or three at a time with 8507R Rack.

8508 Small Tungsten Carbide Grinding Container Small grinding container with puck, dish and lid all made of solid tungsten carbide. Dish has reinforcing aluminum band. 3.6 in. diameter x 2.3 in. high, 7.7 lbs. (9.2 cm x 5.9 cm, 3.5 kg). Nominal volume 80 mL, recommended grinding load 2-20 mL. Includes puck, dish, and lid. Must be used either one or three at a time with 8507R Rack.

8521 Large Hardened Steel Grinding Container Large grinding container with dish, ring, puck, and lid, all made of 52-100 hardened tool steel. 8 in. diameter x 3.3 in. high, 29 lbs. (20.3 cm x 8.2 cm, 13 kg). Nominal volume 650 mL, recommended load 30-100 mL. Includes puck, ring, dish with O-ring gasket and lid.

Typical Suitable for Sample Sample Suitable for Slurry P/N Material Volume Weight Blending Grinding

8501 Hardened Steel 20-50 mL 50 g Y Yes



8504 Tungsten Carbide 20-60 mL 50 g Y Yes

8508 Tungsten Carbide 5-20 mL 15 g Y Yes

8505 Alumina Ceramic 15-40 mL 40 g Y Yes

8506 Zirconia Ceramic 15-40 mL 40 g Y Yes

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Racks and Holders 8507R Rack Holds one or three 8507 or 8508 grinding containers in Shatterbox. Rack base plate has handles to lift Rack into or out of Shatterbox, and four locating pins for positioning one or three grinding containers. Rack lid is matched to Shatterbox clamp, and holds grinding containers down during operation. When 8507R Rack is used with three grinding containers, all three should be the same kind, either 8507 or 8508.

8525 TransporterFor lifting and moving heavier Shatterbox grinding containers (8501, 8504, 8521). Spring-loaded clamp grips lip of dish, twin handles make for easy two-handed lifting.

PrepAid® Grinding Aids & Sample Binders3640 PrepAid Propylene Glycol – (C3H8O2)Liquid grinding aid, 25 grams (approximately 24 mL) in dropper bottle. Instructions included.

3650 PrepAid Vertrel® XFLiquid fluorocarbon grinding aid. Prevents caking, reduces contamination, evaporates after grinding. One-quart bottle. Instructions included.

SPEX SamplePrep 3635 Automated X-Press®

3635-115 Automated X-Press – For 115 V/60 Hz operation

3635-230 Automated X-Press – For 230 V/50 Hz operation CE Approved

• Unique programmable laboratory press

• 35-ton (31.8 metric ton) hydraulic press ideal for repetitive pressing of sample pellets for XRF, IR, and other analytical methods

• Automation improves sample uniformity, consistency and throughput

• Program allows the technician to enter final pressure desired, time to hold at the final pressure, and interval to return back to zero pressure

• Typical pressing cycle is less than two minutes

• Accepts all SPEX SamplePrep die sets

• A safety interlock and automatic pump shut-off safety valve ensure safe operation

Specifications:

Dimensions: 23 in. (58 cm) x 11 in. (28 cm) x 32 ½ in. (82 cm)

Weight: 121 Ibs. net (55 kg); 190 lbs. gross (87 kg)


Motor: 1/3 horsepower


Force: 10–35 tons ram pressure (9.1-31.8 metric tons or 20,000 – 70,000 Ibs.)

Platen Movement: 1.0 inch (2.5 cm)

Platen Size: 3.25 inches diameter (8 cm)

Daylight: Adjustable 2.0 to 6.0 inches (5 to 15 cm)

Upper Screw Adjustment: 3.5 inches (9 cm)

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SPEX SamplePrep 3628 Air-Actuated Bench-Press®

3628 Bench-Press, CE Approved• Air-actuated, manually controlled laboratory press

• 25-ton (22.7 metric ton) maximum pressure is ideal for pressing sample pellets for XRF, IR, and other analytical methods

• Compact design ideally suited for laboratory benchtop applications.

• Patented air-actuation system combines ease of use of a motorized press with the simplicity of a manual press

• Single push-button operation

• Pressure is measured with a dial and the pressure control valve can be set to produce repetitive, reproducible pressures

• Neither hand-pumping nor electrical hook-up is required

• Can be operated off an air compressor or a tank of compressed air

• Requires 90 psi (6.1 atm. or 6.2 bars) and 7 cubic feet per minute (0.2 cubic meters per minute or 198 liters per minute) of air

Specifications:

Dimensions: 12 ½ in. (32 cm) x 16 in. (47 cm) x 30 in. (76 cm)

Weight: 120 lbs. net (55 kg) 205 lbs. gross (93 kg)

Force: 0-25 tons (0-22.7 metric tons or 0-50,000 lbs.)

Platen Movement: 1 ¾ in. (4.4 cm)

Platen Size: 5 ½ in. diameter (14 cm)

Daylight: Adjustable 1 ¼ to 5 ¾ inches (3.2 to 14.6 cm)

3621 SPEX SamplePrep-Carver Model C

Full-size 12-ton (10.9 metric ton) manual hydraulic press with buffer plate accepts all SPEX SamplePrep pellet dies.

Specifications


Weight: 200 Ibs. net (90 kg)

Force: 0–12 tons (0-10.9 metric tons or

0–24,000 Ibs.)

Platen Movement: 5 in. (12.7 cm)

Platen Size: 6 x 6 in. (15 x 15 cm)

Daylight: Adjustable, 0 to 9 in. (0 to 23 cm)

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3626 SPEX SamplePrep-Carver 12-ton Press

Benchtop 12-ton (10.9 metric ton) manual hydraulic press. Ideally suited for IR work with SPEX SamplePrep 3616 - 13 mm Die Set.

Specifications

Dimensions: 20 ¼ in. (51 cm) x 10 ¼ in. (26 cm) x

8 in. (20 cm)


Force: 0–12 tons (0-10.9 metric tons or 0–24,000 Ibs.)

Platen Movement: 5 1/8 in. (13 cm)

Platen Size 4 in. round (10 cm)

Daylight: 5 ¾ inches (14.6 cm)

3622 SPEX SamplePrep-Carver Model M

25-ton (22.7 metric ton) manual hydraulic press with buffer plate accepts all SPEX SamplePrep pellet dies.

Specifications



Force: 0–25 tons (0-22.7 metric tons or 0–50,000 Ibs.)

Platen Movement: 6 ½ in. (16.5 cm)

Platen Size: 9 x 9 in. (23 x 23 cm)

Daylight: Adjustable, 0 to 9 in. (0 to 23 cm)

Accessories

Pellet Die SetsSPEX SamplePrep dies are designed to fit all SPEX SamplePrep presses, and they are equally compatible with other standard laboratory presses using stand-alone dies.

3610 Die Set – 10 mmUsed to form KBr disks for IR analysis. Set includes evacuable die body, base, plunger, two 10 mm polished steel pellets, O-ring vacuum seals, and knock-out ring for sample disk extraction. Load limit 10 tons (9.1 metric tons); weight 2.2 lbs (1 kg).

3613 Die Set – 13 mm Used to form KBr disks for IR analysis. Set includes evacuable die body, base, plunger, two 13 mm polished steel pellets, O-ring vacuum seals, and knock-out ring for sample disk extraction. Load limit 10 tons (9.1 metric tons); weight 2.2 lbs (1 kg).

3623 Die Set – 31 mm To form 31 mm standard sample disks for XRF. Set in cludes evacuable die body, base, plunger, two polished steel pellets, O-ring vacuum seals, and knock-out ring for sample disk extraction. Load limit 50 tons (45.4 metric tons); weight 6.6 lbs (3 kg). Compatible with SPEX SamplePrep 3619 and 3619A Spec-Caps. 31 mm Tungsten Carbide Pellets available (3623C).

3616 Die Set – 35 mm To form 35 mm sample disks. Set includes evacuable die body, base, plunger, two 35 mm polished steel pellets, O-ring vacuum seals, and knock-out ring for sample disk extraction. Load limit 50 tons (45.4 metric tons); weight 8.8 lbs (4 kg). Compatible with SPEX SamplePrep 3615 (33 mm) Spec-Caps. 35 mm Tungsten Carbide Pellets available (3616C).

3614 Die Set – 40 mm To form 40 mm sample disks. Set includes evacuable die body, base, plunger, two 40 mm polished steel pellets, O-ring vacuum seals, and knock-out ring for sample disk extraction. Load limit 50 tons (45.4 metric tons); weight 10 lbs (4.5 kg). Compatible with SPEX SamplePrep 3617 (38 mm) Spec-Caps. 40 mm Tungsten Carbide Pellets available (3614C).

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Replacement Pellets Replacement or spare steel pellets for die operation can be purchased separately. For routine pressing of abrasive materials, SPEX SamplePrep offers 31 mm, 35 mm and 40 mm tungsten carbide pellets which can be substituted for polished steel pellets. Tungsten carbide pellets are much harder than steel, but more brittle, and are not guaranteed against breakage. All replacement pellets are sold in pairs.

3613ST Steel Pellets – 13 mm For 3613 die, sold in pairs.

3623ST Steel Pellets – 31 mmFor 3623 die, sold in pairs.

3623C Tungsten Carbide Pellets - 31 mmFor 3623 die, sold in pairs.


3616C Tungsten Carbide Pellets – 35 mmFor 3616 die, sold in pairs.


3614C Tungsten Carbide Pellets – 40 mmFor 3614 die, sold in pairs.

Sleeve-and-Plunger Sets Sleeve-and-Plunger Sets allow easy preparation of XRF disks with binder matrix supporting a layer of sample. Makes sturdy disks, prevents damage to die; ideal for smaller amounts (<1 g) of analyte. These sets can be purchased separately, or together with SPEX SamplePrep dies at a discount. Please contact SPEX SamplePrep for further information.

3623W Die Sleeve-and-Plunger Set – 31 mmUsed with 3623, 31 mm Evacuable Pellet die and PrepAid Sample Binder. Forms stable XRF sample disks with binder matrix supporting small quantity of sample. Instructions included.

3616W Die Sleeve-and-Plunger Set – 35 mmUsed with 3616, 35 mm Evacuable Pellet Die and PrepAid Sample Binder. Forms stable XRF sample disks with binder matrix supporting small quantity of sample. Instructions included.

3614W Die Sleeve-and-Plunger Set – 40 mmUsed with 3614, 40 mm Evacuable Pellet Die and PrepAid Sample Binder. Forms stable XRF sample disks with binder matrix supporting small quantity of sample. Instructions included.

Spec-Caps® for Reinforcing XRF Sample Disks SPEX SamplePrep Spec-Caps are shallow, thin-walled aluminum cups which are routinely used in the production of pressed-powder sample disks for OES, XRF, and other analytical techniques. The Spec-Cap forms the bottom and sides of the finished pellet. Thus reinforced, sample disks are resistant to chipping and breaking, and are more easily handled, marked, and stored than unclad disks.

3619 Spec-Cap® - 30 mm30 mm wide x 8 mm deep, for 3623 die. Produces pellets 31 mm x 5 mm. Outside painted to prevent seizure in die and to facilitate labeling. Sold in units of 100.

3619M Spec-Cap – 30 mmSame as 3619. Sold in units of 1000.

3619A Pre-Flared Spec-Cap – 31 mm31 mm wide x 8 mm deep, for 3623 die. Produces pellets 31 mm x 5 mm. The 3619A has been pre-flared for a snug fit in the 3623 31 mm Evacuable Pellet Die, and has an unpainted surface. It may be substituted for the 3619 30 mm Spec-Cap in all applications where flaring is recommended. Eliminates need for 3618 Edge-Flaring Tool. Sold in units of 100.

3619AM Pre-Flared Spec-Cap – 31 mmSame as 3619A. Sold in units of 1000.

3615 Spec-Cap – 33 mm33 mm wide x 8 mm deep, for 3616 die. Produces pellets 35 mm x 5 mm. Outside painted to prevent seizure in die and to facilitate labeling. Sold in units of 100.


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3617 Spec-Cap – 38 mm38 mm wide x 9 mm deep, for 3614 die. Produces pellets 40 mm x 5 mm. Outside is painted to prevent seizure in die and to facilitate labeling. Sold in units of 100.


3618 Edge-Flaring Tool For 3617 and 3619 Spec-Caps. This tool widens the Spec-Cap rim to ensure a snug fit against the walls of the die, thus preventing sample loss and pellet jamming.

3625 Edge-Flaring Tool For 3615 Spec-Caps.

SPEX SamplePrep PrepAid® Binders 3642-150 PrepAid Cellulose Binder – (C6H10O5)n≤ 20 µm powder, 150g bottle. Can be blended with sample at 10% by weight to form XRF sample disk, or used undiluted as sample matrix with Sleeve-and-Plunger Set. Will bond 200-250 disks or provide matrix for 25-35 supported sample disks. Instructions included.

3642-450 PrepAid Cellulose Binder – (C6H10O5)nSame as 3642-150, but a 450 g bottle. Will bond 600-750 disks or provide matrix for 72-105 supported sample disks. Instructions included.

3644-150 PrepAid UltraBind®< 20 µm powder, 150g bottle. Can be blended with sample at 10% by weight to form XRF sample disks, or used undiluted as sample matrix with Sleeve-and-Plunger Set. Combines the moisture resistance of Paraffin Binder along with the ease of removing a prepared disk from the die set associated with the Cellulose Binder. UltraBind may include low levels of NaCl. Will bond 200–250 disks or provide matrix for 25–35 supported-sample disks. Instructions included.

3644-450 PrepAid UltraBind®Same as 3644-150, but a 450g bottle. Will bond 600-750 disks or provide matrix for 75-105 supported sample disks. Instructions included.

3646-150 PrepAid Paraffin Binder – (CnH2n+2)≤ 30 µm powder, 150g bottle. Will bond 200-250 disks. Can be blended with sample at 10–20% by weight to form moisture-resistant XRF sample disk. Instructions included.

3646-450 PrepAid Paraffin Binder – (CnH2n+2)Same as 3646-150, but a 450g bottle. Will bond 600-750 disks. Instructions included.

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Katanax® K1 Prime Automated Electric Fluxer

Available in 115 V/60 Hz and 230 V/50 Hz, CE Approved models

• Single-position automated electric fusion machine.

• Prepare glass disks for XRF or solutions for AA, ICP, and wet chemistry analysis.

• Throughput of 5-7 samples/hour.

• Includes built-in and customizable fusion methods.

• Multi-lingual, foolproof fusion method development wizard.

• Independent heaters for the crucible and mold enable optimal temperature control and homogenization.

• Real-time temperature display.

• User-adjustable holding temperature to minimize ramping time.

• Both disk and solution units included.

• Please specify mold size when ordering.

• Solution Agitation Unit available.

Complete K1 Prime Kit includes:

• K1 Prime Fluxer

• Mold Heating/Cooling Unit

• Unbreakable Teflon Beaker

• Mold Holder

• Power Cord

• Instruction Manual

• Tool Kit (hex keys, spare fuses)

Specifications

Voltages 110-127VAC, 220-240VAC (50-60Hz)

Power (max) 1300W

Dimensions 15.6” (39.5cm) x 10.9” (27.5cm) x 23.1” (58.5cm)

Mass: 66 lbs (30kg)

Power Cord 3-prong standard North American plug for 115V / 60 Hz Model 2-prong European CEE 7/7 plug for 230V / 50Hz Model

Katanax® K2 Prime Automated Electric Fluxer

For 230 V/ 50/60 Hz operation, CE Approved

• Six-position automated electric fusion machine

• Prepare glass disks for XRF or solutions for AA, ICP, and wet chemistry analysis

• Throughput of 20-30 samples/hour

• Includes built-in and customizable fusion methods

• Multi-lingual, foolproof fusion method development wizard

• Closed electric oven ensures all crucibles and molds are at the same temperature

• Real-time temperature display

• User-adjustable holding temperature to minimize ramping time

• Both disk and solution units are included

• Please specify mold size when ordering

• Solution kit available, accommodates 5 Teflon® beakers.

Complete K2 Prime Kit includes:

• K2 Prime Fluxer

• Special outlet for installation hook-up

• Tool Kit (hex keys, spare fuses)

• Instruction Manual

Specification:

Voltages 220-240 VAC (50-60Hz)

Power (max) 3000 W

Dimensions 19” (48 cm) x 36” (91 cm) x 19” (48 cm)

Mass 95 lbs (43 kg)

Power Cord 3-prong, 240V standard North American plug

Patent Pending

Patent Pending

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Crucibles and Molds for Katanax K1 and K2 Automated Electric Fluxers

The Katanax K1 and K2 accept crucibles with the following characteristics:

• Capacity - 30 mL

• Straight-walled

• Height – 32 mm

• Outer Diameter – 41 mm

• Made of 95% platinum, 5% gold (borate fusions) or 100% zirconium (peroxide fusions)

• Standard or reinforced

The Katanax K1 and K2 accept common round molds with the following characteristics:

• Depth - 6 mm

• Inner Base Diameter - 30 to 40 mm

• Outer Rim Diameter - 39 to 49 mm

Accessories

SPEX SamplePrep offers crucibles made of 95% platinum and 5% gold, the standard non-wetting alloy for borate fusions. Zirconium crucibles are available for peroxide fusions. Pt/Au crucibles and molds are offered in regular and heavy-duty versions; heavy-duty platinumware lasts longer, has better temperature control, and is less likely to warp. While the Katanax K1 and K2 Automated Electric Fluxers accept most common molds, they require crucibles with very particular specifications. All platinumware is sold separately from the instrument. Price quotations are available upon request by contacting SPEX SamplePrep Customer Service at (732) 623-0465. Please note that prices are quoted daily and may change due to market fluctuations.

Crucibles KP1001A Regular Crucible 95% Pt/5% Au, 26 g. Straight-walled crucible designed specifically for the Katanax K1 and K2 Automated Electric Fluxers. Capacity 30 mL.

KP1002A Heavy-Duty Crucible 95% Pt/5% Au, 30 g. Straight-walled crucible designed specifically for the Katanax K1 and K2 Automated Electric Fluxers with a thicker rim and bottom for greater durability. Capacity 30 mL.

KP0049A Zirconium Crucible 100% Zirconium, 55 g. Straight-walled crucible designed specifically for the Katanax K1 and K2 Automate Electric Fluxers. Capacity 30 mL.

Racks & Holders7151R Rack For holding six 7152 or 7152HP graphite crucibles.

Accessory Packages & Kits7151 Rack and Tongs For holding six 7152 or 7152HP graphite crucibles. Ni-Cr (nichrome) alloys used for rack withstand temperatures as high as 1200° C.

Platinumware, Labware & Handling Tools7151T Long TongsFor handling crucibles; 20 in.

7154 Short TongsFor handling crucibles; 9 in.

Molds & Mold Holders KP1003A Mold – 30 mm95% Pt/5% Au, 18 g.

KP1004A Heavy-Duty Mold – 30 mm95% Pt/5% Au, 30 g.

KP1005A Mold – 32 mm95% Pt/5% Au, 21 g.


KP1007A Mold – 35 mm95% Pt/5% Au, 25 g.

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KP1009A Mold – 40 mm95% Pt/5% Au, 34 g.

KP1010A Heavy-Duty Mold – 40 mm 95% Pt/5% Au, 48 g.

KP1011A Platinum Crucible Heater Lid 95% Pt/5% Au, 16 g. Platinum crucible heater lid used in place of the zirconium crucible heater lid that is supplied with the K1. Recommended for users concerned with possible contamination due to the zirconium lid.

KP1015A Zirconium Crucible Heater Lid100% Zirconium crucible heater lid as supplied with the K1. Protects the crucible, conserves heat during fusion, and aids in pouring.

KP0010A Teflon® BeakerUnbreakable Teflon beaker used for preparing solutions.

Mold Holders for the K1 Prime Automated Electric Fluxer KP0037A Mold Holder, for 30 mm molds

KP0038A Mold Holder, for 32 mm molds



Mold Holders and Plates for the K2 Prime Automated Electric FluxerAll Mold Plates are designed to hold six molds. Custom sizes are available.

KP0235A Mold Plate, for 35 mm molds

KP5603A – Mold holder, 30mm, 6-position




Graphite Crucibles for Muffle Furnaces 7152 Graphite Crucibles 31.8 mm O.D. x 28.6 mm, 8.4 mL capacity. Regular purity graphite; sold in units of 100.

7152HP Graphite Crucibles31.8 mm O.D. x 28.6 mm, 8.4 mL capacity. High purity graphite; sold in units of 10.

7153 Graphite Crucibles Micro, 9.5 mm O.D. x 19 mm, 0.6 mL capacity. High-purity graphite; sold in units of 100.

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7155 Graphite Crucibles 38.1 mm O.D. x 46.0 mm, 20 ml capacity, 25.4 mm I.D. flat bottom. High-purity graphite; sold in units of 10.

7156 Graphite Crucibles 38.1 mm O.D. x 46.0 mm, 20 ml capacity, 25.4 mm I.D. flat bottom. Regular-purity graphite; sold in units of 10.

7157 Graphite Crucibles For casting 31 mm glass disks, 44.4 mm O.D. x 38.1 mm, 27 ml capacity, 31 mm I.D. flat bottom. High-purity graphite; sold in units of 10.

7158 Graphite CruciblesFor casting 31 mm glass disks, 44.4 mm O.D. x 38.1 mm, 27 mL capacity, 31 mm I.D. flat bottom. Regular-purity graphite; sold in units of 10.

7159 Graphite Crucibles For casting 33.7 mm glass disks, 50.8 mm O.D. x 38.1 mm, 32 mL capacity, 33.7 mm I.D. flat bottom. High-purity graphite; sold in units of 10.

7160 Graphite CruciblesFor casting 33.7 mm glass disks, 50.8 mm O.D. x 38.1 mm, 32 mL capacity, 33.7 mm I.D. flat bottom. Regular-purity graphite; sold in units of 10.

7161 Graphite Crucibles For casting 40 mm glass disks, 57.1 mm O.D. x 38.1 mm, 44.5 mL capacity, 40 mm I.D. flat bottom. High-purity graphite; sold in units of 10.

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7162 Graphite CruciblesFor casting 40 mm glass disks, 57.1 mm O.D. x 38.1 mm, 44.5 mL capacity, 40 mm I.D. flat bottom. Regular-purity graphite; sold in units of 10.

NOTE: Glass disks cast in graphite crucibles will have rough faces which should be polished flat for analytical accuracy.

Fusion Flux SPEX SamplePrep is proud to offer the full line of SPEX Fusion Fluxes and additives. These fluxes have exceptional qualities due to their purity and coarse glass-bead texture.

Some of these fluxes include lithium bromide or lithium iodide as a non-wetting agent. A small amount of LiBr or LiI is added to the flux when making a “glass bead” (lithium borate sample disk), to keep the melt from sticking to the crucible or mold. Additional LiBr or LiI is added to the flux before making a solution for ICP/AA analysis, so the entire melt is transferred to the beaker. LiBr solution in a dropper bottle is a convenient way to add non-wetting agent to a fusion, at 6-7 mg per drop.

Please call SPEX SamplePrep for a free sample of flux, or information on a custom flux.

SPEX Fusion Flux Mixture Composition (%) Pure Ultra-Pure

LiT 100.00 FFB-1000-02 FFB-1000-03

LiT/LiBr 99.50/0.50 FFB-1005-02 FFB-1005-03

LiT/LiI 99.50/0.50 FFB-1007-02 FFB-1007-03

LiT/LiM 67.00/33.00 FFB-6700-02 FFB-6700-03

LiT/LiM/LiBr 66.67/32.83/0.50 FFB-6705-02 FFB-6705-03

LiT/LiM/LiI 66.67/32.83/0.50 FFB-6707-02 FFB-6707-03

LiT/LiM 50.00/50.00 FFB-5000-02 FFB-5000-03

LiT/LiM/LiBr 49.75/49.75/0.50 FFB-5005-02 FFB-5005-03

LiT/LiM/LiI 49.75/49.75/0.50 FFB-5007-02 FFB-5007-03

LiT/LiM 35.00/65.00 (12/22) FFB-3500-02 FFB-3500-03

LiT/LiM/LiBr 34.83/64.67/0.50 (11.94/21.89/0.17) FFB-3505-02 FFB-3505-03

LiM 100.00 FFB-0000-02 FFB-0000-03

LiM/LiBr 99.50/0.50 FFB-0005-02 FFB-0005-03

LiM/LiBr 98.50/1.50 FFB-0007-02 FFB-0007-03

AdditivesAdditive Description Part Number

LiBr crystal (125g) FFB-100-03

LiBr – 15mL solution FFB-103-03

LiBr – 15mL solution (10pk) FFB-105-03

LiI crystal (125g) – trihydrate salt FFB-110-03

LiI – 15mL solution FFB-113-03

LiI – 15mL solution (10pk) FFB-115-03

LiF crystals (125g) FFB-200-03

LiNO3 (250g) FFB-300-03

LiNO3 (500g) FFB-301-03

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XRF Accessories

Introduction The XRF spectroscopist is confronted with an almost infinite variety of samples. Liquid samples, from water to oil, require virtually no preparation. Many solid samples, if homogeneous, can be machined to the proper shape and run directly. Others, such as refractories, ores, and biological materials, are too inhomogeneous, and must be pulverized or fused. Then the homogenized material can be dissolved for analysis as a liquid sample, pressed into pellet form, or cast as a glasslike solid.

SPEX SamplePrep manufactures disposable sample-handling accessories for powder and liquid XRF samples. Over the last twenty years, millions of powder samples for XRF have been pressed and protected in the SPEX SamplePrep Spec-Cap, an aluminum cap for reinforcing pellets. Millions more liquid XRF samples have been run in SPEX SamplePrep X-Cells. These and the related products listed here will save you time and money, and assure you of reproducible, contamination-free results.

Spec-Caps® SPEX SamplePrep® Spec-Caps are shallow, thin-walled aluminum cups which are routinely used in the production of pressed-powder sample disks for OES, XRF, and other analytical techniques. The Spec-Cap forms the bottom and sides of the finished pellet. Thus reinforced, sample disks are resistant to chipping and breaking, and are more easily handled, marked, and stored than unclad disks.

3619 30 mm Spec-Cap®30 mm wide x 8 mm deep, for 3623 die. Produces pellets 31 mm x 5 mm. Outside painted to prevent seizure in die and to facilitate labeling. Sold in units of 100.

3619M 30 mm Spec-CapSame as 3619. Sold in units of 1000.

3619A 31 mm Pre-Flared Spec-Cap31 mm wide x 8 mm deep, for 3623 die. Produces pellets 31 mm x 5 mm. The 3619A has been pre-flared for a snug fit in the 3623 31 mm Evacuable Pellet Die, and has an unpainted surface. It may be substituted for the 3619 30 mm Spec-Cap in all applications where flaring is recommended. Eliminates need for 3618 Edge-Flaring Tool. Sold in units of 100.

3619AM 31 mm Pre-Flared Spec-CapSame as 3619A. Sold in units of 1000.

3615 33 mm Spec-Cap 33 mm wide x 8 mm deep, for 3616 die. Produces pellets 35 mm x 5 mm. Outside painted to prevent seizure in die and to facilitate labeling. Sold in units of 100.


3617 38 mm Spec-Cap38 mm wide x 9 mm deep, for 3614 die. Produces pellets 40 mm x 5 mm. Outside is painted to prevent seizure in die and to facilitate labeling. Sold in units of 100.


3618 Edge-Flaring Tool For 3617 and 3619 Spec-Caps. This tool widens the Spec-Cap rim to ensure a snug fit against the walls of the die, thus preventing sample loss and pellet jamming.

3625 Edge-Flaring Tool For 3615 Spec-Caps.

This tool widens the Spec-Cap rim to ensure a snug fit against the walls of the die, thus preventing sample loss and pellet jamming.

PrepAid Binders 3642-150 PrepAid Cellulose Binder – (C6H10O5)n≤ 20 µm powder, 150 g bottle. Can be blended with sample at 10% by weight to form XRF sample disk, or used undiluted as sample matrix with Sleeve-and-Plunger Set. Will bond 200-250 disks or provide matrix for 25-35 supported sample disks. Instructions included.

3642-450 PrepAid Cellulose Binder – (C6H10O5)nSame as 3642-150, but a 450 g bottle. Will bond 600-750 disks or provide matrix for 72-105 supported sample disks. Instructions included.

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3644-150 PrepAid UltraBind®< 20 µm powder, 150 g bottle. Can be blended with sample at 10% by weight to form XRF sample disks, or used undiluted as sample matrix with Sleeve-and-Plunger Set. Combines the moisture resistance of Paraffin Binder along with the ease of removing a prepared disk from the die set associated with the Cellulose Binder. UltraBind may include low levels of NaCl (See Section 2, Chapter 3). Will bond 200–250 disks or provide matrix for 25–35 supported-sample disks. Instructions included.

3644-450 PrepAid UltraBind®Same as 3644-150, but a 450 g bottle. Will bond 600-750 disks or provide matrix for 75-105 supported sample disks. Instructions included.

3646-150 PrepAid Paraffin Binder – (CnH2n+2)≤ 30 µm powder, 150 g bottle. Will bond 200-250 disks. Can be blended with sample at 10–20% by weight to form moisture-resis tant XRF sample disk. Instructions included.

3646-450 PrepAid Paraffin Binder – (CnH2n+2) Same as 3646-150, but a 450 g bottle. Bonds 600-750 disks. Instructions included.

X-Cells3527 X-Cell, 40 mm Closed X-Ray Cell Polyethylene snap-ring and cup with snap-post vent and reservoir. Requires window film. Sold in units of 100.

3527I Bubble-Free Cell Insert Patented insert for the 3527 40 mm X-cell. Prevents trapped air from forming a bubble under the window film when the X-cell is used in the upright position. Sold separately from 3527 X-Cell in units of 100. Instructions included. For normal optics XRF spectrometers made by most major manufacturers.

3529 X-Cell, 31 mm Closed X-Ray CellPolyethylene snap-ring and cup with snap-post vent and reservoir. Requires window film. Sold in units of 100.

3561 X-Cell, 31 mm Universal Closed X-Ray Cell with CollarPolyethylene snap-ring, collar, and cup with snap-post vent and reservoir. Squared snap-ring design insures proper positioning in standard 31 mm sample holders. Requires window film. Sold in units of 100.

3565 X-Cell, 43 mm Closed X-Ray Cell For Horiba Sulfur Analyzers. Polyethylene snap-on collar and cup with vent and reservoir. Features recessed window, and fill mark on inside of cup. Requires window film. Sold in units of 100.

3571 X-Cell, 31 mm Double Open End X-Ray Cell with Collar Two polyethylene snap-rings, collar, and open body. Versatile design accepts liquids, pastes, powders and other solids. Squared snap-ring insures proper positioning in standard 31 mm sample hold ers. Can be run with open back, sealed window back, or micro-porous film. Requires window film. Sold in units of 100.

3577 X-Cell, 31 mm Micro X-ray Cell with Collar Two polyethylene snap-rings, collar, and open body. Unique cell for small, air-sensitive or hazardous samples. 6.3 mm window is centered in standard 31 mm sample holder. Can be run with open back, sealed window back, or micro-porous film. For hazardous or sensitive samples, top can be fitted with a standard 7 x 13 mm serum bottle closure for injection, purging, or sample isolation. Requires window film. Sold in units of 100.

SPEX SamplePrep X-Cells

Note: All measurements in mm unless otherwise noted

X-Cell Overall Overall Window Capacity Number Description Height O.D. Plane O.D. Aperture (mL)

3529 31 mm X-Cell 22.0 32.0 30.5 24.5 8.0

3561 31 mm Universal X-Cell 22.0 31.6 31.6 24.5 8.0

3571 31 mm Open End X-Cell 22.0 31.6 31.6 24.5 10.0

3527 40 mm X-Cell 22 39.5 38.0 31.5 13

3565 43 mm X-Cell 19 48.0 43.0 38.8 11.5*

3577 Micro X-Cell 22 31.6 31.6 6.3 0.5

*To fill line

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SPEX SamplePrep X-Cells are designed to fit the major brands of XRF spectrometers such as PANalytical, Rigaku, Bruker, and Thermo Instruments. The requirements of the application will dictate the type and size of X-Cell and which window material to use.

Thin Film3511 Kapton® Window Film (Roll) 0.3 mil (8 µm) thick. Supplied as continuous roll 2 7/8 in. (73 mm) wide, 50 ft. (15 m) long, in box with serrated edge for tearing film to desired length.

3512 Kapton Pre-Cut Circles0.3 mil (8 µm) thick. Supplied as pre-cut circles 2 ½ in. (63.5 mm) diameter, 500 sheets per box.

3515 Mylar® Pre-Cut Circles (Large) 0.25 mil (6 µm) thick. Supplied as pre-cut circles 3 ¼ in. (82.6 mm) diameter, 500 sheets per box.

3516 Mylar Window Film (Roll) 0.12 mil (3 µm) thick. Supplied as continuous roll 2 7/8 in. (73 mm) wide, 300 ft. (92 m) long, in box with serrated edge for tearing to desired length.

3517 Mylar Window Film (Roll)0.25 mil (6 µm) thick. Supplied as continuous roll 2 7/8 in. (73 mm) wide, 300 ft. (92 m) long, in box with serrated edge for tearing film to desired length.

3518 Mylar Pre-Cut Circles 0.25 mil (6 µm) thick. Supplied as pre-cut circles 2 ½ in. (63.5 mm) diameter, 500 sheets per box.

3520 Polypropylene Window Film (Roll) 0.2 mil (5 µm) thick. Supplied as continuous roll 2 7/8 in. (73 mm) wide, 300 ft. (92 m) long, in box with serrated edge for tearing film to desired length.

3521 Polypropylene Pre-Cut Circles 0.2 mil (5 µm) thick. Supplied as pre-cut circles 2 ½ in. (63.5 mm) diameter, 500 sheets per box.

3524 Micro-porous Teflon® Film (Roll) Film has 0.2 µm pores. Equalizes pressure when installed on back of 3571 and 3577 X-Cells. Not a window film. Supplied as continuous roll 2 in. (51 mm) wide, 17 ft. (5 m) long in box with serrated edge for cutting film to length.

3525 Ultralene® Window Film (Roll) 0.16 mil (4 µm) thick. Supplied as continuous roll 2 7/8 in. (73 mm) wide, 300 ft. (92 m) long, in box with serrated edge for cutting film to desired length.

3526 Ultralene® Pre-Cut Circles 0.16 mil (4 µm) thick. Supplied as pre-cut circles 2 ½ in. (63.5 mm) diameter, 500 per box with paper interleaving.

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Thickness Thickness Film P/N Description (mil) (micron) Type Qty Diameter

3511 Kapton .3 8 Roll 50 ft (15m) -

3512 Kapton .3 8 Precut 500 Circles 2 ½ in.(63.5mm)

3515 Mylar .25 6 Precut 500 Circles 3 ¼ in (82.6mm)

3516 Mylar .12 3 Roll 300 ft (92m) -

3517 Mylar .25 6 Roll 300 ft (92m) -

3518 Mylar .25 6 Precut 500 Circles 2 ½ in (63.5mm)

3520 Polypropylene .20 5 Roll 300 ft (92m) -

3521 Polypropylene .20 5 Precut 500 Circles 2 ½ in (63.5mm)

3524 Micro-Porous Teflon .20 Micro-Porous Roll 17 ft (5m) -

3525 Ultralene .16 4 Roll 300ft (92m) -

3526 Ultralene .16 4 Precut 500 Circles 2 ½ in (63.5mm)

Certified Reference Materials for Organic and Inorganic Spectroscopy and Chromatography

SPEX CertiPrep® has been providing Certified Reference Materials (CRMs) for spectroscopy and chromatography for over fifty-five years. Our products are manufactured in Metuchen, NJ, and used by laboratories around the world.

As new technologies have developed, we made it our business to produce superior Certified Reference Materials for those instruments and technologies. We have a full line of inorganic and organic standards for AA, ICP, ICP-MS, IC, GC, GC/MS, HPLC and LC/MS.

We built our reputation on providing chemists and environmental scientists with products that exhibit Quality, Reliability and Convenience. Our Web site offers online ordering capabilities, product searches, and a technical knowledgebase. Our SPoints program rewards our valued customers with SPoints that are redeemable for great gifts.

To request the latest SPEX CertiPrep CRM literature, visit our Web site at www.spexcertiprep.com, or contact CRM customer service via email at [email protected] or via phone at either 732-549-7144 or 1-800-LAB-SPEX, x444.

Services available from SPEX CertiPrep

Extended Customer ServiceTechnical Customer Service is available 8:00 am - 5:30 p.m. (EST) Monday - Friday.

Total Customer SatisfactionSPEX CertiPrep has always been committed to providing every customer with “Total Customer Satisfaction.” We take pride in ensuring every customer is satisfied with our products and services. Our support does not end when the product reaches the customer’s facility; we have technical support available to answer any question.

New SPEX CertiPrep websiteSPEX CertiPrep’s easy-to-navigate Web site was designed to give our customers easier access to our large product offering and technical knowledgebase. Customers can view catalog products and pricing, place orders, and submit requests for custom standards. Registered users can also check order status, download Certificates of Analysis, and view their current SPoints balance. Our Web site also offers technical whitepapers, posters, and presentations. Visitors can contact us through the Web site’s “Live Chat” control for instant order and technical assistance. Visit us at www.spexcertiprep.com today!

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AccreditationsSPEX CertiPrep is certified by UL DQS for ISO 9001 and accredited by A2LA as complying with the requirements of ISO/IEC 17025 and ISO/IEC Guide 34. These certifications and accreditations cover both our organic and inorganic Certified Reference Materials, giving us the most comprehensive scope in the industry.

SPEXertificate® SPEX CertiPrep’s SPEXertificate® is the most comprehensive Certificate of Analysis in the industry! The SPEXertificate® includes actual reported values for the final solution – not reported values of the starting materials or by a calculation. The certificate also includes trace impurity measurements for up to 68 elements, measurement uncertainty, and NIST traceability when available.

Products available from SPEX CertiPrep

Custom Standards SPEX CertiPrep offers a comprehensive custom Certified Reference Materials program to fit the needs of each customer. Custom standards are designed around the requirements of our customer’s individual methods or procedures. Our chemists evaluate the custom standards request and determine elements/analytes compatibility and standard matrix needed to keep the product stable. We have an established database of chemical interactions from our years of standard manufacturing experience to help ensure every custom standard we product is of the highest quality.

We offer custom standards for most of our inorganic product lines in volumes of 100 mL, 250 mL, 500 mL and 1L. Organic customs are available in ampules from 1 to 20 mL. We can recommend standards that will work with specific methods and prevent interferences. Contact us 1-800-LAB-SPEX or www.spexcertiprep.com for further information.

Assurance® Grade Standards for ICP and AASPEX CertiPrep’s Assurance® Grade Single Element Standards are high quality calibration standards for both AA and ICP instrumentation. Each Assurance® Grade Single Element standard comes with a Certificate of Analysis that lists the values of the main element measured by two independent methods, and reports actual measured impurities of up to 30 elements.

Assurance® Multi-Element Solution StandardsSPEX CertiPrep offers a large catalog of multi-element standards formulated for the most common environmental analyses. Stock standards include a number of blends containing metals of concern in groundwater, drinking water and solid waste.

Assurance® Contract Laboratory Program (CLP) StandardsCLP standards are a complete series of multi-element standards and blanks designed for use in the U.S. Environmental Protection Agency’s Contract Laboratory Program, or CLP. These solutions provide

everything needed for instrument calibrations, calibration verifications, interference checks, calibration blanks, and sample spikes. The standards are to be used in conjunction with the US EPA’s Statement of Work for Inorganic Analysis; Multi-Media/ Multi-Concentration Document Number ILM06.0/ILM05.2.

Claritas PPT® Grade Standards for ICP-AES and ICP-MS. SPEX CertiPrep Claritas PPT standards are a class of certified inorganic reference standards designed specifically for today’s new generation of trace ICP-AES and ICP-MS instrumentation. Our chemists have formulated this line of high-purity standards for user convenience and stability, while providing documentation tailored to extremely low levels of detection. Every Claritas PPT Standard is supplied with a comprehensive SPEXertificate® of Analysis that reports actual measured values in the solution of both the major analytes and up to 68 trace element impurities at PPT levels.

Ion ChromatographyThe Certificate of Analysis for every Ion Chromatography Standard reports ICP trace metal analysis of the starting materials, classical wet assay values for the major constituents, and an ion chromatography check for the major ion(s). SPEX CertiPrep Eluents are made from high purity salts and filtered ASTM Type I water.

Ion Selective Electrode StandardsIon Standards are compatible with all of the leading Ion Selective Electrodes and ionic strength adjustment buffers and are made from high purity salts and filtered ASTM Type I water.

Organometallic Oil Standards SPEX CertiPrep presents a comprehensive offering of 37 single-element standards at 1000 µg/g and 5000 µg/g in base oil; popular 23-, 21-, 12- and 5-element blends in base oil; sulfur in #2 diesel fuel; sulfur oil standards for diesel fuel analysis in base oil; and base oil and kerosene blanks. These standards are commonly used for the determination of wear metals in engine oils and other lubricants in machines such as automobiles, aircraft, heavy equipment, trucks, locomotives, and military vehicles.

Consumer Safety StandardsSPEX CertiPrep offers a line of Certified Reference Materials for a range of Consumer Safety Products analysis. We offer standards designed for the analysis of metals in plastic toys, Phthalates in polyethylene, and RoHS/WEEE analysis.

Organic Reference Materials for GC, GC/MS, HPLC & LC/MSEach standard is sold in 1 mL ampules and comes with a pre-labeled “transfer/storage” vial for extending shelf life. The concentration of each analyte in the standard is optimized, taking into account the vapor pressure, evaporation rates, breakdown rates and dilution schemes. SPEX CertiPrep offers stock standards used with environmental testing methods for 500 Series Drinking Water Methods, 600 Series Wastewater Methods, and 8000 Series Solid and Hazardous Waste Methods.

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Contamination Control ProductsMade for Chemists by Chemists®. Our line of contamination control products are designed by SPEX CertiPrep’s chemists in response to the need for cost-effective, easy-to-use equipment and high-purity matrix/wash blanks for the clean laboratory environment. Our Pipette Washer/Dryer is lightweight, compact, convenient to use, and made of durable polyethylene. Independent valves control flow of water to the back and front rows of washer. The unit contains 23 cone-shaped plastic pipette holders which are designed to accommodate pipettes from 0.5-250 mL in size. An optional pump and basin are also available.

Sub-boiling Acid Stills are available in PTFE or Quartz; these Acid Stills allow the preparation of ultra-pure acids right in the laboratory at a fraction of the cost of purchasing them ready-made.

Solids and Matrix Based Reference MaterialsSPEX CertiPrep offers a wide range of Solid and Matrix based Certified Reference Materials to customers worldwide. We provide over 8000 RMs and CRMs sourced from over 100 producers across the globe.

CRMS and RMs for:• Metal – Disks • Oil & Coal• Metal – Chips • Food & Agriculture• Ferro-Alloys • Environment & Health• Geological Samples • Biological Fluids• Refractory Samples • Pure Chemicals• Industrial By-Products/Process • Physical Testing

Please visit our website for more details: www.spexcertiprep.com.

TechnicalInformation

Section 2


Sec tion 2

Chapter 1

Tissue Homogenization / Cell Lysis

Chapter 1


Sec tion 2Tissue Homogenization / Cell Lysis

SPEX SamplePrep Freezer/Mills are cryogenic laboratory mills which chill samples in liquid nitrogen and pulverize them with a magnetically driven impactor. Each sample is placed in a separate grinding vial which is immersed in a liquid nitrogen bath inside the mill. Thus there is no cross-sample contamination, and the low temperature of the sample is maintained during grinding. These features have made the SPEX Freezer/Mill the most effective in the world, the “mill of last resort” for many normally ungrindable samples.

Freezer/Mill technology, field-proven since the 1960s, has been built into three models. All models include significant safety features, and electronic controls which allow an entire grinding sequence to be programmed.

Bead Milling

Bead milling is the preferred method to disrupt a variety of microorganisms and plant or animal tissues. In bead milling, grinding media such as steel or ceramic balls or glass beads are vigorously agitated inside a sealed vial or titer plate with the sample. The most commonly used grinding balls are steel ball bearings of 4mm in diameter. Disruption or cell lysis occurs as a result of the crushing action of the grinding media as they collide with the sample. Low shearing of nucleic acids can be achieved by varying the agitation speed of the mill. It is considered the method of choice for disruption for yeast and fungi and tough-to-disrupt cells like bacteria and microalgaew.

The method is one of the few that totally avoids possible cross-contamination between samples because both vials and grinding media are disposable. SPEX SamplePrep offers a comprehensive range of grinding media, vials and titer plates for samples ranging from 0.6mL up to 50mL. The size and amount of the grinding media used is important. Speed and effectiveness of disruption can be increased dramatically by increasing the density, form and amount of grinding media in the sample vial or titer plate. Tough tissues may require precooling to embrittle the sample and this also serves to preserve any temperature sensitive components such as RNA or Proteins. SPEX SamplePrep offers a full range of cryogenic, Kryo-Tech® accessories for this purpose. The loading of the beads should always allow adequate movement inside the sample vials but can be up to 80% of the total sample volume, provided there is still adequate agitation of the bead slurry. The higher the ratio of grinding media to sample volume, the faster the rate of cell disruption. After homogenization, the beads settle and the cell extract can be removed.

The Geno/Grinder® offers analysts a versatile high-throughput bead mill for plant or animal tissue homogenization. The patented design with a true linear grinding motion provides the most efficient mechanism for tissue homogenization and cell lysis. The powerful and compact design enables complete disruption with or without buffer in about 1-3 minutes with high yields. The Geno/Grinder® has also been successfully used for pesticide residue extraction from fruit and vegetable samples using the QuEChERS technique.

Details of different extraction techniques and appropriate grinding media for different sample types can be found in the application notes section of this handbook.

Cryogenic Grinding

Cryogenic grinding of plant and animal tissue is a very effective technique for the Microbiologist. Samples that require extraction of nucleic acids must be kept at -80º C or lower during the entire extraction process. Utilizing SPEX SamplePrep Freezer/Mills will maintain the sample temperatures well below these critical temperatures during the automated grinding sequence. For samples that are soft or flexible at room temperature, cryogenic grinding may be the only viable technique for processing your samples. A detailed explanation of the various cryogenic techniques can be found in Section 2, Chapter 2.

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Chapter 2


Grinding & Pulverizing / Mixing & Blending

Chapter 2

89Order Online 24/7: www.spexsampleprep.com

Sec tion 2Grinding & Pulverizing / Mixing & Blending

Chart A – SPEX SamplePrep Sample Container Materials Selection Chart

This chart reviews the key properties of each grinding container material, including major and minor elements, hardness, resistance to abrasion, durability, and efficiency. These are usually the basis for selecting the proper container for a particular application.

Chart B – Compostition of Standard Steels Used in SPEX SamplePrep Equipment and Accessories

This chart is essentially an extension of Chart C that lists all the steel alloys used in SPEX SamplePrep grinding containers and accessories, with the theoretical composition of each given in percentages.

To discuss any SPEX SamplePrep mills and containers, and their suitability for your applications, feel free to contact our product specialists. We also offer test-grinding of your samples in our applications lab, or we can loan you a mill for short-term trials in your own lab. The more you know about SPEX SamplePrep mills, the more confident you can be that they will do the job you need to get done.

The recommended maximum sample volume for grinding is approximately 20% of a container’s volume, for blending 40–50%. Optimum grinding efficiency is achieved with samples toward the middle of the volume range.

The “typical grinding sample” is quartz sand, with a density of approximately 1.3g/mL, in a volume conducive to efficient grinding. Clearly the analyst must choose the proper weight for his sample, based on its density and other characteristics.

When to Pulverize

The nonhomogeneity of materials in the real world presents a serious sampling dilemma: how to represent a large, often nonuniform whole with a small analytical sample. Often the best approach is first to take a quantity of the material large enough to be compositionally representative and reduce it to a fine homogeneous powder. Then the sample can be adapted to a particular analytical technique, and used as is, pressed into a sample disk, fused, dissolved, etc.

SPEX SamplePrep has laboratory mills and grinding containers capable of pulverizing and blending all kinds of samples, from metals to plastics, rocks to living tissue, and pesticides to pharmaceuticals.

How to Select a Laboratory Mill and Grinding Container

SPEX SamplePrep mills are intended for the analytical laboratory. They are not for batch or production milling, but for rapid, efficient grinding of analytical samples ranging in amounts from less than one gram up to approximately 100 grams. The hallmark SPEX SamplePrep Freezer/Mills, Mixer/Mills and Shatterbox all have separate grinding containers for individual samples, so after use the container is cleaned and not the mill. SPEX SamplePrep mills were designed for spectroscopists who were concerned not only about reducing samples to uniform homogeneous powders, but also about what was added to the samples during processing.

The information that follows addresses these concerns and can help you to select the proper SPEX SamplePrep mill and grinding container for your application.

If your samples can be pulverized by impact at room temperature, you have a choice of several proven SPEX SamplePrep mills. The 8500 and 8530 Shatterboxes are ring-and-puck mills that are ideal for rapid grinding of samples up to approximately 100 grams, in 5 minutes or less. The 8000M and 8000D Mixer/Mills are high-energy ball mills that not only pulverize samples in the 10 gram range but are suitable for blending powders and making emulsions up to approximately 60 mL. The 5100 Mixer/Mill is a smaller high-energy ball mill, with about 10% the sample capacity of the 8000-series Mixer/Mills. All these mills have multiple-sample capacity with smaller samples, and a wide choice of grinding container sizes and materials, for maximum on-the-job flexibility over a wide range of applications.

The 6770, 6870 and 6970 EFM Freezer Mills are unique cryogenic mills that can grind almost any “ungrindable” sample, including all plant and animal tissue, most polymers, and countless other samples that are resistant to room-temperature milling. Freezer/Mills require liquid nitrogen for chilling the sample, and maintaining cryogenic temperatures in the vial during grinding. Forty years after its introduction the Freezer/Mill remains the most widely used and most effective cryogenic mill in the world.

The following charts are available to assist you in your selection of a SPEX SamplePrep mill and grinding container, or containers. (If your lab deals with a variety of samples, more than one type of container may be necessary).

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Chapter 2



Chart A: SPEX SamplePrep Container Materials Selection Chart

Material Major Minor Hardness Resistance Durability Comparitive Element(s) Element(s) to Abrasion Efficiency

Hardened Steel Fe Cr, Si, Mn, C Mohs: 5 ½ - 6 Moderate High High Rockwell:C 60-65

Stainless Steel Fe, Cr Ni, Mn, S, Si1 Mohs: 5 – 5 ½ Moderate High High Rockwell: C 55-60

Cr-Free Steel Fe C, Mn, Si, Mo Mohs:5 – 5 ½ Moderate High High Rockwell: C 55-60

Tungsten Carbide W, C, Co Te, Ti, Nb Mohs: 8 ½ + Knoop: High Long-wearing Very High 1400-1800 Subject to breakage

Alumina Ceramic Al Si, Ca, Mg Mohs: 9 Very High Long-wearing Moderate Rockwell: R45N 83 Brittle Knoop: 2100

Agate Si Al, Na, Fe, K, Ca, Mg1 Mohs: 6 – 7 Extremely Very long- Moderate Knoop: 550 – 800 High wearing

Zirconia Zr Mg, Hf Mohs 8 ½ Extremely Very long- Moderate Rockwell: R45N 74-79 High wearing Knoop 1160

Silicon Nitride Si Y, Al, Fe, Ca Mohs 8 ½ + Extremely Very long- Moderate Knoop 1600 High wearing

Plastic C - Mohs 1 ½ Low Low Low for Disposable grinding; High for blending

1 All reported less than 0.02%Ch

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Chapter 2



Limits of Cryogenic Grinding

Not every substance which is difficult to grind at room temperature can be pulverized by cryogenic milling. For this reason we at SPEX SamplePrep recommend that you discuss your samples and grinding requirements with one of our specialists before selecting a mill. If there is any question about whether a sample can be ground in a Freezer/Mill we will request that a portion be submitted for test-grinding.

Nearly every naturally occurring material of biological origin can be cryogenically ground, usually to a fine particle size: hair, muscle tissue, bone, wood, plant stems and roots, seeds, cotton fiber, etc.

Polymers in general can be ground, but their physical form is important. Many flexible plastics which can routinely be milled in pellet form are difficult to grind as thin fibers and films, which remain flexible even at -200°C. For the same reason, polymers in general cannot be ground to as fine a particle size as biological samples. As the particles become smaller they become more flexible, and progressively more difficult to grind. Many silicone compounds remain elastic and hard-to-grind in any form.

The behavior of metals and metal alloys during cryogenic milling is highly variable. For example, impure copper as well as many copper alloys can be ground without much trouble, while pure copper remains malleable. Metal samples as a rule should be test-ground.

When a sample initially resists grinding, it may not be because it is “ungrindable.” Many such samples respond to the strategies of reducing the sample volume, increasing grinding time, and even lengthening the pre-cooling period to 30 minutes or so. Successes have also been scored by using “filler” grinding agents such as detergent and quartz sand.

Guide to Freezer/Mill Sample Capacity

The optimum volume, weight, grinding time, and impact frequency for any sample ground in a SPEX SamplePrep Freezer/Mill are determined by experimentation, the experience of the operator, and the requirements of the analyst. As a rule of thumb, the smaller the sample and the longer it is ground, the finer the particle size will be. In cryogenic grinding, temperature also affects the outcome: the colder a sample, the more finely it can be ground.

The 6751 Vial has a sample volume limit of about 5 mL; bulkier samples may restrict movement of the impactor. For efficient grinding, typical sample volume is 1 to 2 mL, corresponding to weights of 1 to 2 grams for polymers and 2 to 4 grams for bone. Specialized grinding vials with similar capacity are the 6761 Poly-Vial for metal-free grinding of delicate samples, and the 6771 Cr-Free Vial for grinding electronic components without adding chromium.

The 6753 Microvial set includes three 6753V Microvials. Each Microvial can hold a small amount of sample, typically between 30 mg and 200 mg.

The 6801 Vial has at least 50 mL maximum sample capacity, 10 times that of the 6770 Freezer/Mill. Typical sample volumes are 10 to 20 mL, equivalent to weights of 10 to 20 grams for polymers and 20 to 40 grams for bone. The new series of Mid-Size Vials has about half the capacity of the 6801 vial, and

Key Points About Cryogenic Grinding

While cryogenic grinding has long been a vital sample preparation tool for the analytical chemist, it rarely receives publicity. The recent identification of the remains of Czar Nicholas II of Russia not only solved a mystery nearly eighty years old but also emphasized the importance of cryogenic techniques in both forensic and archaeological research. DNA was successfully extracted from the Romanov bone fragments after they had been ground in a SPEX SamplePrep Freezer/Mill.

Many analytical samples which are too flexible or sensitive to be impact-ground at room temperature can be embrittled by chilling, and then pulverized. These include polymers, rubber, textiles, cereal grains, hair, fingernails, skin, bone, and muscle tissue. There are also many samples which degrade in various ways during normal grinding, but whose critical properties are preserved by chilling. Coal, for example, can be cryogenically ground to retain its more volatile components, and clay minerals may be pulverized for XRD study without distorting their crystal structure. Bone, fingernails, and other biological materials can be cryogenically ground in preparation for nucleic acid extraction without damaging DNA and even RNA through heating.

Cryogenic grinding presents unique challenges for designers of laboratory mills, as the low temperatures required are very hard on mechanical equip ment. The SPEX SamplePrep Freezer/Mills impels a steel impactor magnetically while the sample vial is immersed in liquid nitrogen. Chilling times vary with the mill and the insulating ability of the sample but can range from 10 minutes to 30 minutes or more. Once chilled, samples can be pulverized quickly; grinding times rarely exceed a few minutes with either mill.

As with samples ground at room temperature, those processed cryo genically vary enormously in their “grindability,” and it is difficult to predict the optimum sample size, chilling time, or grinding time. In addition, a given sample material may exist in several forms, some of which are easy to grind while others are very difficult. For example, polypropylene is supplied to molders as small pellets, but can be made into thin films or fibers; the pellets grind fairly easily, as they become rigid when chilled, but the films and fibers may remain flexible and become difficult to grind cryogenically.

We strongly suggest submitting samples for test grinding if there is any question about the suitability of our mills and techniques for your application.

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Free Grinding Vial (6771, 6871, or 6883). This vial also contains a magnetically driven Cr-free impactor. The vial is then loaded into the Freezer/Mill and immersed in liquid nitrogen until the contents are thoroughly chilled, usually a matter of 10-15 minutes, but as much as 20-30 minutes for complicated components such as the USB flash drive and RAM module. The sample inside the vial is pulverized, and because the sample is isolated in a closed grinding vial, cross-sample contamination is easily controlled and sample integrity is maintained. Grinding time can vary greatly depending on the type of sample and its pliability but a grinding time of 8-10 minutes can be expected in the case of the USB flash drive and RAM module. The resultant fine, uniform powder (Fig. 2) can then be used for XRF, fusion, or other types of analyses.

We now offer a range of powder Polymers, Solder and Glass standards to come designed for RoHS WEEE compliance limits. Please contact our European Division, SPEX CertiPrep Ltd. for additional information.

includes a Poly-Vial and Cr-Free Vial as well as the standard 6881 Mid-Size Vial. The 6870 Freezer/Mill can also hold up four 6751, 6761, and 6771 vials for simultaneous grinding of multiple smaller samples.

Cryogenic Grinding for RoHS/WEEE

The Restriction of Hazardous Substances (RoHS) and Waste Electrical and Electronic Equipment (WEEE) Directives of the European Union were introduced to minimize the accumulation of hazardous waste in landfills from the disposal of electrical and electronic equipment. The concentrations of hazardous substances such as lead, cadmium, mercury, chromium VI, polybrominated diphenylethers (PBDEs), and polybrominated biphenyls (PBBs) are restricted in electrical and electronic products and/or components.

RoHS/WEEE states that if the component can be mechanically separated, then each component is subject to the RoHS limits. The definition of exactly what this means is an ongoing process. However, one thing is certain; in order to get an accurate analytical result, these products and components must be reduced to homogeneous, representative samples. Many components such as circuit boards, wire, solder, polymers and resins are difficult if not impossible to grind using traditional methods. Cryogenic grinding in the SPEX SamplePrep Freezer/Mill® is the easiest way to homogenize these materials.

Consider analyzing circuit boards: for example, USB flash drives that act as portable hard disk drives, and the larger RAM modules that are installed on computer motherboards.

These samples (Fig. 1) are composites of many materials: metals, resins, substrates, etc. A small piece taken from a board here or there at random would not be representative of the whole board, much less an entire batch of boards. Hence, sampling many boards, and in effect homogenizing them, assures that the sample used for analysis is representative of the batch. However, memory boards are made to be rugged and durable over a wide range of conditions, and are very difficult to grind into powder. Furthermore, some of the components may be more fragile than others. Metals, resins and other materials all have distinct and different properties when put through conventional laboratory mills. Some components may grind at room temperature, and some may not. Some may require a shearing action and other abrasion. When all of these components are part of the same integral memory board, it is impractical to break down the board into its several components, each suited for their own special type of mill.

For such situations, cryogenic grinding in the SPEX SamplePrep Freezer/Mill is the answer. The general principle behind cryogenic grinding is to chill samples until they are brittle, and then break them up through impact, crushing, or shearing. In the case of the Freezer/Mill, the sample, such as the USB flash drive or RAM module, would be first cut into manageable pieces. These are then placed in the appropriately sized SPEX SamplePrep Chromium-

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Chapter 2



Key Points About Grinding

Grinding Container Materials

Your selection of appropriate grinding and mixing containers is as important as your choice of a mill. The proper grinding vial or dish will greatly enhance analytical accuracy. The grinding container is generally harder than the material to be ground and should be of a substance whose presence in the sample will not interfere with analysis. SPEX SamplePrep’s containers have been selected with care to offer optimum capability in the lab.

Methacrylate and polystyrene vials and balls are ideal for pulverizing soft, brittle materials and for mixing and storing powders. With these vials only traces of organic impurities are added to your sample.

Hardened steel is durable and tough, suitable for general-purpose grinding; it’s the workhorse of the lab. When hard substances are ground in steel containers, some Fe and Cr contamination can be expected. Stainless steel is less subject to chemical attack, but contributes Ni as well. Cr-free steel rusts easily but can be used to grind samples for RoHS/WEEE testing. Halide-releasing compounds corrode steel and should be ground in tungsten carbide, alumina, agate, zirconia, or silicon nitride containers.

Tungsten carbide vials and dishes are the most effective and versatile of all. Tungsten carbide is substantially harder and heavier than steel, grinding is faster and contamination is minimal. Other than tungsten, cobalt (a binder) is the major contaminant. If cared for, tungsten carbide containers will last indefinitely. They’re recommended as a long-term “best buy” for grinding almost anything.

Alumina ceramic is ideal for extremely hard samples or in cases where steel and tungsten carbide contaminants are objectionable. All SPEX SamplePrep alumina components are 99.5% pure aluminum oxide, with some silicon, calcium, and magnesium present. It is lightweight and brittle but very abrasion-resistant. Necessary for an important minority of samples, alumina ceramic vials are a helpful addition to any lab’s grinding armory.

Agate is harder than steel, and chemically inert to almost anything except HF. It’s also brittle and must be handled with care. Agate vials are for the grinding and mixing of samples when organic and metallic contamination is equally undesirable. Agate is 99.9% silica and is extremely wear-resistant.

Silicon nitride is a tough space-age material with remarkable wear characteristics, and hardness superior to agate and zirconia. If it is important to have a container whose only major contaminant is silicon, consider SPEX SamplePrep silicon nitride. It is extremely durable compared to agate, and while it contains some yttria and alumina, overall contamination levels will be very, very low.

Zirconia is a ceramic which in many ways approaches the ideal grinding medium. Since it is both hard and tough it wears very slowly, adding little contamination. It is about one and one-half times as dense as alumina, grinding almost as fast as steel. And because it is mostly zirconium oxide with low percentages of magnesium oxide and hafnium oxide, the contamination SPEX SamplePrep zirconia ceramic does contribute is often not important to the analyst.

Grinding Tests with the Freezer/Mill

Material Form Weight (g) Time* Final Size Mesh

Aluminum Foil 2 mil Piece 11 3 x 2 100-200

Black loaded thermoplastic Pellets 3 3 x 1 100-200

Candle Wax Chunk 1.5 2 100-200

Cardboard Corrugated 0.5 2 x 2 200

Fish Scales 10mm Flakes 1.5 2 x 2 200

Hair Dog Hair Clipping 0.5 2 200

Hot Melt Adhesive Chunk 0.5 2 x 2 50

Human Tooth Whole Tooth 2 1 x 2 200

Human Vertebrae 7mm Pieces 2 2 x 2 200

Mouse Skin Raw, ½ Animal 22 3 200

Nylon3 3mm Beads 2 2 x 24 100-200

Permalloy 5 2mm Shot 2 3 30

Polyethylene 10mil Sheet 1 2 200

Polypropylene Fibers 1.5 3 x 2 100-200

Rubber ½ in. Thick Sheet 0.5 2 x 2 50-100

Rubber, Oil-Extended 5mm Shearings 1.55 2 25-50

Sheep Wool Wad 0.5 2 200

Space Food Stick 2 2 100

Teflon 2mil Tape 3 2 x 2 100

Duct Tape Tape 0.5 2 x 2 100

Urethane Elastomer Pellets 1 3 x 1 100-200

Wood Chips 1.3 2 x 2 100

*3 x 2 denotes grinding periods of 2 minutes each with re-cooling of approximately 4 minutes between grinds1) 0.5g of Tide detergent was added2) Equal weight of sodium sulfate was added as dehydrating agent.3) Three different nylons yielded similar results4) 2x – two-minute grinds with one-minute cooling period between5) Equal amount of sand added; purpose: ethanol-toluene extraction

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Dry grinding is simpler and quicker, but requires much more careful matching of the technique to the sample. If the caking is due to moisture, as in many soils and cements, the sample can be dried before grinding. Other samples can be successfully ground with a variety of additives. Dry soaps/detergents are lubricants, and some also include an abrasive; graphite is an anti-static agent as well as a lubricant; there are many proprietary grinding aids as well, which may contain an abrasive, a lubricant and a binding agent. Other grinding aids include polyvinyl alcohol, phenyl acetate and aspirin.

Howard Kanare of Construction Technology Laboratories, Inc. has publicized the use of propylene glycol (one drop for up to ten grams of sample, roughly 0.3 wt.%) for laboratory fine grinding of Portland cement and many minerals. In a swing mill such as the SPEX SamplePrep Shatterbox, oven-dried samples can be ground quickly to less than ten microns without agglomeration or sticking to the mill walls. Propylene glycol must be used safely, after consulting the material safety data sheet; it is available as a PrepAid product.

Vertrel® XF, a DuPont product sold as a cleaning fluid, is finding increased acceptance as a grinding aid. A fluorocarbon fluid, it prevents sample caking during grinding, and quickly evaporates from an open grinding container without leaving any residue. Our experience is that the “grindability” of almost any sample is enhanced by the use of Vertrel XF, contamination is lowered, and the grinding container is easier to clean. Typical XRF samples such as cement, rock, clinker, and similar material can be routinely ground below 10 microns. Vertrel XF also lowers contamination levels from the grinding container, and is available as a PrepAid product.

The technique that follows was pioneered by John Anzelmo and colleagues at Bruker AXS and is here adapted for SPEX SamplePrep equipment: in an 8501 Hardened Steel Grinding Container load together 10 grams of sample, 2.5 grams of 3642 Cellulose or 3644 Ultrabind binder, and 7 mL of Vertrel XF. Grind in an 8530 Shatterbox for 2.0 minutes, open the grinding container in a hood until the Vertrel XF has evaporated. Prepare a 3614 40mm Pellet Die with a flared 3617 Spec-Cap, and transfer the ground sample/binder powder to the die. Press at 20 tons for 0.3 minutes in a 3635 X-Press.

Reference

Anzelmo, John; Seyfarth, Alexander; and Arias, Larry (2001).

Approaching a Universal Sample Preparation Method for XRF Analysis of Powder Materials. Advances in X-ray Analysis, Vol. 44, JCPDS – International Centre for Diffraction Data, Newtown Square PA.

Cleaning Containers

Grinding containers should be cleaned between sample runs to avoid cross contamination, and the procedure can be as simple or as complex as your analytical objectives warrant. In some applications a simple wipedown with ethanol may suffice; another practical approach is to brush out a container, then briefly grind an expendable portion of the next sample and discard it.

Contamination

Processing a sample always contaminates it. Successful analysis depends on recognizing the sources of contamination and controlling them. When the contaminants are known and can be quantified, analytical results can be refined accordingly.

As the grinding container is the major source of contamination, its selection is critical. In general, one’s objective should be to minimize contamination levels while avoiding elements which will interfere with analysis. An example is the grinding of steel slags in a tungsten carbide container: tungsten carbide grinds rapidly, and the expected low-level contaminants of tungsten, carbon, and cobalt are not generally looked for in these slags. SPEX SamplePrep’s selection of grinding container materials gives you maximum flexibility in choosing the best approach for your samples and analytical aims.

Major, minor, and trace elements predictably found in SPEX SamplePrep grinding containers are listed in Chart A, Section 2, Chapter 2. However, strictly speaking, almost no two grinding containers will have exactly the same elemental profile. There are many different steels, carbides, and ceramics, each with specific compositions. Often the formulas are proprietary, so that a type of tungsten carbide engineered to have specific properties will have a different makeup from two different manufacturers. In addition, there are inevitable variations from batch to batch of the same material, both in the exact proportions of the major elements and in trace element composition.

Because of these variations in grinding container composition, we strongly recommend determining the exact elemental profile of your individual grinding containers, preferably with your own analytical equipment and techniques. The simplest approach is to grind samples of known composition and see what is added by grinding. Lacking known samples, one may grind portions of a single sample for increasing lengths of time, and check to see which elements increase in proportion to grinding time. Once the contributed impurities and their proportions are known for a grinding container, the resulting profile can be fitted to the analytical results, regardless of the actual contamination level. (While this level is important, it clearly will vary with the composition and condition of the grinding container, the size, hardness, and toughness of the sample, and grinding time).

Grinding Aids

When samples agglomerate or “cake” during grinding, further particle size reduction is clearly inhibited. Caking can result from moisture, heat, static charge accumulation, the fusing of particles under pressure, and other causes. Many of the techniques which make sample preparation an “art” are devoted to getting around caking, and we can only hint at the possibilities.

Slurry grinding is an obvious approach; if particles remain in suspension during grinding, they are unlikely to cake. Water, alcohol, or other liquids are added to the sample before grinding, and removed afterwards. Although slurry grinding is a reasonably reliable way of grinding a sample to micron-sized particles, it is sloppy and time-consuming, requires a leak-proof grinding container, and adds extra steps to one’s sample preparation procedure.

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Mixer/Mill, widely known as the “SPEX Mill”, has become the industry standard for mechanical alloying applications. The high energy of the milling action, and the durability of the motor, allow running for extended periods.

Over the years nearly one hundred articles have been published in refereed scientific journals regarding the SPEX SamplePrep Mixer/Mill and its use for mechanical alloying. This includes mechanical alloying techniques, evaluations of grinding vial materials, and numerous other topics. The following publication list is intended to highlight the more recent publications that we are aware of and is not intended to be comprehensive. If you are considering the Mixer/Mill for your own mechanical alloying application, we strongly encourage you to do your own search for applicable publications and references.

Effect of the heating rate on crystallization behavior of mechanically alloyed Mg50Ni50 amorphous alloy. Aydinbeyli, N., Nuri Celik, O., Gasan, H., Aybar, K. International Journal of Hydrogen Energy, Vol. 31, Issue: 15, December, 2006. pp. 2266-2273.

Effect of ball milling on simultaneous spark plasma synthesis and densification of TiC-TiB2 composites. Locci, A.M., Orru, R., Cao, G., Munir, Z.A. Materials Science and Engineering A, Vol. 434, Issue: 1-2, October 25, 2006. pp. 23-29.

Temperature of the milling balls in shaker and planetary mills. Takacs, L., McHenry, J. S. Journal of Materials Science, Vol. 41, Issue: 16, August 2006. pp. 5246 – 5249.

Modeling of comminution processes in Spex Mixer/Mill. Concas, A., Lai, N., Pisu, M., Cao, G. Chemical Engineering Science, Vol. 61, Issue: 11, June, 2006. pp. 3746-3760.

Effect of mechanical alloying conditions on the microstructure evolution and electrode characteristics of Mg63Ni30Y7. Khorkounov, B., Gebert, A., Mickel, Ch., Schultz, L. Journal of Alloys and Compounds, Vol. 416, Issue: 1-2, June 8, 2006. pp. 110-119.

A study of mechanical alloying processes using reactive milling and discrete element modeling. Ward, T.S., Chen, W., Schoenitz, M., Dave, R.N., Dreizin, E.L. Acta Materialia, Vol. 53, Issue: 10, June, 2005. pp. 2909-2918.

Microstructural evolution during mechanical alloying of Mg and Ni. Rojas, P., Ordonez, S., Serafini, D., Zuniga, A., Lavernia, E. Journal of Alloys and Compounds, Vol. 391, Issue: 1-2, April 5, 2005. pp. 267-276.

Mechanical milling of magnesium powder. Hwang, S., Nishimura, C., McCormick, P.G. Materials Science and Engineering: A, Vol. 318, Issue: 1-2, November, 2001. pp. 22–33.

Formation of supersaturated solid solutions by mechanical alloying. Huang, B.-L., Perez, R.J., Lavernia, E.J., Luton, M.J. Nanostructured Materials, Vol. 7, Issue: 1-2, January 2, 1996. pp. 67-79.

Synthesis of nanocrystalline Fe-B-Si powders. Perez, R.J., Huang, B.-L., Crawford, P.J., Sharif, A.A., Lavernia, E.J. Nanostructured Materials, Vol. 7, Issue: 1-2, January 2, 1996. pp. 47-56.

For more thorough cleaning one may grind one or more batches of pure quartz sand, and then wash the container thoroughly. In extreme cases, such as the plating of container walls with a malleable metal, chemical cleaning or multiple grinds with quartz may be necessary.

An effective single-step grinding procedure for most grinding containers is to grind pure quartz sand together with hot water and detergent, then rinse and dry the container. Drying is speeded by the use of a blow-dryer or similar appliance. A safety advantage of this cleaning method is that it controls respirable airborne dust.

A cleaning procedure is easily evaluated by grinding and analyzing a known sample, or even by checking the impurities appearing in successive batches of ground quartz sand. It should be noted that grinding containers become more difficult to clean with age because of progressive pitting and scratching of the grinding surfaces.

Hardened steel and even stainless steel containers can rust. While iron oxide coatings can be removed by warm dilute oxalic acid solution or abrasive cleaning, we recommend that steel containers be thoroughly dried after cleaning and, if stored, kept in a plastic bag with a desiccating agent.

Respiratory Protection

The general objective of sample grinding is, of course, to convert an inhomogeneous solid to a fine, homogeneous powder. Inevitably, some of this powder is released into the environment, usually during the emptying or cleaning of the grinding container. We strongly recommend the wearing of an approved dust-mask during this procedure, and suggest the use of a laboratory fume hood. Even harmless rock and cement samples can become potentially harmful to the respiratory system when finely pulverized.

Infrared Mulls

KBr pellets and Nujol mulls are quickly prepared with SPEX SamplePrep Mixer/Mills using small plastic (3111), agate (3118) and steel vials (3114). The polystyrene spectrum appears as a constant background and can easily be subtracted. Steel and agate produce no IR background.

Pelletizing Samples

Powder can be turned into pellets for X-ray and IR analysis with SPEX SamplePrep presses and dies. We offer 12-ton and 25-ton manual presses, the 25-ton air-actuated 3628 Bench-Press, and the 35-ton 3635 Automated X-Press.

Mechanical Alloying

Mechanical alloying, also referred to as reactive milling, is a process originally developed for the production of oxide dispersion strengthened super alloys. Today, mechanical alloying is often used as a solid-state powder processing technique that generates powders with unique microstructures. A high-energy ball mill can be used to accomplish this. Over the past few decades, the SPEX SamplePrep

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8000 Series Mixer/Mills

The SPEX SamplePrep 8000 Series Mixer/Mills are efficient compact laboratory mills capable of pulverizing one or two samples in the 10-gram range. There are two versions: the classic 8000M Mixer/Mill with one clamp, and the 8000D Mixer/Mill with two clamps.

The single clamp 8000M (previously the 8000) is known simply as “the SPEX Mill” to thousands of users and has been in service for over forty years. The clamp mechanism has changed little since the earliest versions, simply because it has proven extremely durable. Many of the earliest 8000 Mixer/Mills are still in service. Now the wind-up timer has been replaced by an electronic 100-minute timer (extended timer available) and the mill’s safety features have been improved.

The dual-clamp 8000D Mixer/Mill combines two clamps on one shaft with an electronic 100-minute timer (extended timer available). As the two clamps move in balance, vibration is reduced and component life extended. A fan keeps the clamps and motor cool during operation.

Applications8000 Series Mixer/Mills have been used for pulverizing rocks, minerals, sand, cement, slag, ceramics, catalyst supports, and hundreds of other brittle, often hard samples. Early on, the Mixer/Mill was used to grind samples, and then blend them with graphite for arc/spark spectroscopy. Now similar samples are ground and then blended with binder before being pressed into samples disks for XRF. The vigorous motion of the clamp is also excellent for making emulsions such as paints, inks, and pharmaceuticals. The 8000M Mixer/Mills have also achieved notoriety for their ability to mechanically alloy small quantities of materials.

Grinding Vials8000 Series grinding vials are 2 ¼ in. (5.7 cm) wide and up to 3 in. (7.1 cm) long. They have an internal volume of 50-60 mL with a grinding capacity of about 10 mL and a blending capacity of 25 mL. They are available in hardened and stainless steel, tungsten carbide, alumina ceramic, zirconia ceramic, silicon nitride, agate, and methacrylate.

We recommend that vials for the 8000D Mixer/Mill be purchased and used in pairs to maintain the balanced motion of the clamps. If only one sample needs to be ground, the second similar vial should be clamped in place without the sample or grinding ball.

Grinding ActionThe SPEX SamplePrep 8000 Series Mixer/Mills are functionally described as shaker mills or high-energy ball mills. They shake one or two containers back and forth several thousand times a minute. In each clamp, the vial, which contains a sample and one or more balls, is shaken in a complex motion which combines back-and-forth swings with short lateral movements, each end of the vials describing a “figure-eight”. The length of that swing is the same as the internal length of the vial, about two inches

SPEX SamplePrep Mixer/Mills

5100 Mixer/Mill

The SPEX SamplePrep 5100 is a highly effective impact grinder and blender for samples weighing 1 gram or less and offers many advantages over other small mixers.

ApplicationsThe 5100 Mixer/Mill is ideal for grinding small amounts of brittle samples such as minerals, rocks, or glass. It has even been used for grinding gallstones. The 5100 Mixer/Mill can also be used for mixing powders and slurry grinding and is commonly used to blend samples with KBr for IR analysis.

Grinding VialsThe continuously adjustable jaws accept vials from 1 in. (25.4 mm) to 2½ in. (63.5 mm) long and up to 3/4 in. (19 mm) in diameter without adapters. It holds three 1/2 in. (12.7 mm) diameter or two 3/4 in. (19 mm) diameter vials. Vials are available in plastic (polystyrene and polycarbonate), hardened and stainless steel, agate, and tungsten carbide. All of the vials except for the agate are able to accommodate slurry grinding.

Grinding ActionThe high grinding and mixing efficiency of the 5100 Mixer/Mill is a result of its three-dimensional action. A one-inch (25.4 mm) component of motion along the axis of the vial is complemented by two motions at right angles to the vial axis: a 3/16 in. (4.8 mm) horizontal movement and a ¼ in. (6.35 mm) vertical oscillation. The resulting action is consistent with the general shape and size of the vials. That is, at every stroke there is an impact at the end of the vial – over 100 per second – to crush the material rapidly and reproducibly.

ControlsThe 115 V/60 Hz version of the 5100 Mixer/Mill has a push-button, resettable 30-minute timer. The 230 V/50 Hz version has a push-button, resettable 72-minute timer.

Safety FeaturesThe rugged construction of the 5100 Mixer/Mill ensures a long life of heavy work. The steel housing protects the entire clamp mechanism and vials and the lid is equipped with a locking latch.

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Pulverizing and Blending Grinding Tests with 8000 Series Mixer/Mills

Material Form Vial – Time (min) Amount (g) % Passing Method 325 mesh

Antimony Pieces L-D 5 26 97Asbestos Fluff WC-D 10 * *Bauxite 60 mesh HS-W 30 3 *Bismuth Chunks PJ-D 20 5 75Bone Chunks AC-D 10 * 35Boron Carbide Chunks WC-D 15 7 100Brake Linings Chunks WC-D * * *Carbon (activated) Pieces HS-D 10 10 90Carbnauba Wax Piece PJ-D 2 5 20Cement (Portland) Powder AC-W 30 20 100Chrome Ore Chunk WC-D 10 15 39Chromium Chunk WC-W 20 10 50Cobalot Pieces WC-W 10 10 91Copper Shot WC-D 15 2 95Ferro Cr 100 mesh WC-W 20 5 94Ferro Nb - WC-W 60 5 10Floor Tile Chunk WC-D ** ** **Germanium Pieces L-D 5 5 38Ilmenite Grains WC-D 10 5 98Limonite Ore Grains HS-W 30 3 100Porcelain Chunk WC-D 15 6 83Potassium Pyrosulfate Fused Button PV-D 10 5 100Reforming Catalyst 3mm Beads AC-D 5 5 *Sand Grains WC-D 2 12 86Silica Chips L-D 30 15 *Silica Chips AC-D 20 5 97Silicon Chunks WC-D 15 10 92Silicon 6mm Lumps L-D 10 5 30

(5.08 cm). With each swing the balls impact against one end of the vial, simultaneously milling the sample to a powder and blending it. Due to the amplitude and velocity of the clamp’s swing, each ball develops fairly high G-forces, enough to pulverize the toughest rocks, slags, and ceramics.

ControlsThe electronic timer of the 8000 Series Mixer/Mills displays the programmed running time in minutes and seconds. While the mill is operating, the timer also counts down the amount of time left in the run. Push-button controls include start, stop and pause functions as well as timed programming. The timer is factory-set for a maximum of 100 minutes but this may be extended to 1,000 or 10,000 minutes for special applications such as mechanical alloying with the installation of a special chip. Prolonged continuous operation of Mixer/Mills requires special maintenance and warranty restriction may apply. Contact SPEX SamplePrep for details.

Safety FeaturesThe 8000 Series Mixer/Mill’s rugged and durable construction is the reason why many of the original mills are still in use. A steel cabinet protects the entire clamp mechanism and vials. Both of the 8000 Series Mixer/Mills are equipped with safety interlocks so that they cannot be operated with the lid open. Each clamp has a lock nut to prevent it from loosening while the mill is running. The motors are also equipped with a thermal overload protector and the 230-volt 8000M and 8000D Mixer/Mills are CE Approved.

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SPEX SamplePrep Shatterbox

Since its introduction over thirty years ago, the SPEX SamplePrep Shatterbox has become the most popular “swing mill” in the United States. Now as then, it is the most efficient way to pulverize up to 100 grams of brittle material to analytical fineness.

There are two Shatterbox models, the full-sized, 8530 Enclosed Shatterbox, and the original 8500, which offers the same grinding performance as the 8530 but without the timer and cabinet.

ApplicationsShatterboxes typically grind cement mix, rocks, slags, soils, ceramics, and ores, but have been used for hundreds of other materials including sulfur pellets, dried marsh-grass, and pharmaceuticals. The original SPEX SamplePrep swing mill, the 8500 Shatterbox, was developed for cement plants and steel mills with open, noisy lab conditions and is still used in many locations where the working environment requires a rugged, simple mill. The 8530 Shatterbox is ideal for labs concerned with minimizing noise and maximizing safety.

Grinding ContainersThe standard grinding containers, with a nominal volume of about 100 mL, is available in hardened steel (8501), tungsten carbide (8504), alumina ceramic (8505), and zirconia ceramic (8506). Typical grinding loads for samples such as cement, rock, or slag are 50 grams for the steel and tungsten carbide containers and 30 grams for the ceramic containers. Note that the actual capacity of the containers is much higher, and (for example) the steel container has been used for as little as 4 grams and as much as 100 grams. However, grinding efficiency decreases as the container becomes more tightly packed.

There are also small grinding containers made of hardened steel (8507) and tungsten carbide (8508), which can be run either one or three at a time with the 8507R Rack. These grinding containers have about a fifth the capacity of the standard grinding containers and a typical sample size of 10 grams.

An oversize steel grinding container (8521) is also available and has approximately a 150 mL capacity. This corresponds to a typical sample size of 75-100 grams. The 8525 Transporter can be used to move the heavier grinding containers (8501, 8504, 8521) in and out of the Shatterbox.

Grinding ActionThe Shatterbox swings a dish-shaped grinding container in a tight, high-speed horizontal circle. Inside the container are the sample, a puck, and (in most containers) a ring. As these grinding elements swing free inside the container, the sample is rapidly crushed between the walls of the container, puck, and ring, and further reduced by the millstone-like action of the puck and ring against the container floor. Grinding times are usually between two and five minutes, with resultant particle size well below 200 mesh (approximately 50 microns). With grinding aids, smaller samples can be reduced below 10 microns.

(con’t) Pulverizing and Blending Grinding Tests with 8000 Series Mixer/Mills

Material Form Vial – Time (min) Amount (g) % Passing Method 325 mesh

Slag (blast furnance) - HS-W 20 3 100Slag (copper) 100 mesh WC-W 10 5 84

Slag (open hearth) - HS-W 20 3 100

Straw - HS-D 10 5 **

Tomato Stems - HS-D 10 5 **

Transite Chunks WC-D * * *

Tungsten Carbide - WC-W 15 10 100

Tungsten Lumps WC-D 10 25 50

Welding Flux - WC-W 30 5 82

Wood Pieces AC-D 10 1 50

Zirconium Carbide - AC-W 30 15 100

Table Reference

HS – 8001 Hardened Steel Vial L – 8006 Acrylic VialWC – 8004 Tungsten Carbide Vial W – Wet ground (water or trichloroethylene slurry)PJ – 8002 Polystyrene Vial D – Dry groundAC – 8003 Ceramic Vial PV – 6133 Polystyrene Vial*- Suitable for X-ray or optical emission spectroscopy **- Satisfactory for extractions

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Grinding Tests with the 8501 and 8504 Grinding Containers

8501 8504 % % Form as Time Amount Passing Time Amount PassingMaterial Received (min) (g) 325 mesh (min) (g) 325 mesh

Asbestos Fibrous 12 20 100 - - -

Cement (Portland raw mix) >60 mesh 2.5 40* 100 - - -

Ferro-chromium >100 mesh 5 25 100 - - -

Ferro-maganese >200 mesh 3 25 100 - - -

Ferro-molybdenum <80 mesh 4 25 100 - - -

Ferro-niobium <80 mesh 3 25 100 - - -

Ferro-silicon <80 mesh 4 25 100 - - -

Fero-titanium <80 mesh 6 25 100 - - -

Fiberglass Thin sheets 2 10 100 2 15 100

Flourite >100 mesh 3 50 100 - - -

Garnet (synthetic) Chunks 2 200 15 10 200 96

Glass Chunks 6 35 100 2 35 100

Graphite Mineral Fiber Fiber 2 15 100 - - -

Insulation Fiber 4 15 100 - - -

Oil Shale 6.4mm 3 60 100 - - -

Pesticide <100 mesh 15 50 100 - - -

Phosphate, raw mix <60 mesh 2.5 40 100 - - -

Iron Powder <80 mesh 6 5 68 - - -

Sand <10 mesh 10 100 100 4 100 100

Slag, blast furnace Chunks 1 10** 100 - - -

*Sodium alkylarylsulfonate added, 5%**Household detergent (Tide) added, 10%

ControlsThe 8500 Shatterbox can be plugged into a standard lab timer that is not included with the mill. The 8530 Shatterbox features a LCD display with an environmentally safe, push-button membrane switch.

Safety FeaturesThe 8500 Shatterbox contains a motor protected by a perforated shield to allow ample air-cooling for continuous running, and a rubber “skirt” shields the drive mechanism but is easy to move aside when the drive belts need to be replaced. It is shipped with a rubber-matted plywood base to prevent it from walking during operation. The old screw-down clamp has been replaced with the cam-activated clamp to accommodate all SPEX SamplePrep grinding containers.

The 8530 Shatterbox includes a sound-insulated steel cabinet with lockable casters, with which the mill can be moved about the lab and then fixed in place. Safety features of the 8530 Shatterbox include a safety interlock and gas cylinders for the lid. The interlock holds the lid closed whenever the mill is running, and the cylinders control movement of the lid when it is closed or opened.

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Sec tion 2Pressing & Pelletizing

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There are many different procedures for preparing powdered samples for XRF analysis. Typically, however, a representative quantity of the sample is first pulverized, then split to obtain enough powder for an XRF sample disk, usually 6 to 10 grams. That powder is blended with a binder if necessary, and placed in a pellet die (with or without a Spec-Cap) to be pressed into a sample disk which will hold together and has a flat, compositionally uniform surface. This disk is then placed in the sample holder of the XRF spectrometer.

An alternate technique, particularly useful when only a small amount of sample is available, incorporates a thin layer of sample on a disk of binder. The SPEX SamplePrep Sleeve-and-Plunger set, used with the appropriate pellet die, makes this procedure easy.

In the end, an exact procedure must be developed for each type of sample. The sampling method and the amount of sample to be ground, the type of mill and the grinding time, what size of die and what pressure to use, whether to include a binder or press the disk in a Spec-Cap, all these details may vary and must be worked out by the analyst to suit his or her samples, equipment, and analytical requirements. Running a series of standards or known samples will help to confirm that your chosen procedure results in accurate, reproducible data.

Particle Size Effects

It is impossible to say a priori how fine to pulverize a given sample. It has been known for many years that the X-ray fluorescence intensity from a sample will increase as the particle size of the sample is decreased. This is due to the reduction in the size and extent of the voids in the sample. By the same reasoning, as the particle size of one of two sample components is decreased, it will yield a higher intensity relative to the component of fixed particle size. Further, if the particle size of both components is decreased, their respective intensities may increase or decrease depending on their relative absorption coefficients. Fortunately, when the particle size becomes small enough, the intensities stabilize.

As an X-ray enters a pelletized sample disk the exciting radiation is absorbed by the matrix. When a particle within the matrix absorbs radiation, the resulting fluorescence is also absorbed, in whole or in part, by the matrix. Hence there is a limit to the depth to which the existing radiation can penetrate and result in fluorescence emitted from the sample. This depth is usually called the critical or infinite thickness.

As particle size becomes small relative to the critical thickness, fluorescence intensities emitted from different sample components approach stable values. This is shown graphically on the next page. In case A, the average penetration of the X-ray is of the same order of magnitude as the large particles, and a change in the size of these particles would have a substantial effect on the fluorescent intensity. This results from the filling of the voids by components with smaller particle sizes. In case B, since the average penetration depth covers many particles of many components, changing the particle size should have little or no effect.

Introduction

Analytical spectroscopic methods such as XRF, OES, and IR often require samples in the form of round, flat-surfaced disks. Although sample disks can be cast from a fusion melt, (refer to section on Fusion Products beginning in Section 2, Chapter 4), they often begin as a powder and are pressed to shape in a pellet die.

The following how-to suggestions apply chiefly to XRF sample preparation, but the basic principles of forming sample disks are the same for OES, spark ablation, IR, etc. The major differences are the diameters of the disks, and the nature of the binders or additives. IR disks are 13 mm across and consist largely of pure potassium bromide (KBr) with the pulverized sample blended in; no Spec-Cap-type jacket is used, as the sample disk must be able to transmit infrared light. OES and spark ablation systems generally use 31 mm disks. As the disk must be electrically conductive for these techniques, most samples are blended with 50% pelletizing-grade graphite, and often pressed in an aluminum Spec-Cap. The diameter of XRF sample disks is dependent on the size of the spectrometer sample holder, which may be 31 mm, 35 mm, 40 mm, or even larger. XRF disks do not require a binder or Spec-Cap if the sample coheres under pressure, but most analysts use a binder or a Spec-Cap or both. The prime requirement for an XRF binder is that it does not contribute impurities. XRF binders include cellulose, paraffin, graphite, orthoboric acid, polyvinyl alcohol, and proprietary products with special properties, e.g. Ultrabind.

For preparation of pellet samples, SPEX SamplePrep offers a full range of laboratory presses, pellet die sets, and die accessories. Presses include a 12-ton and 25-ton SPEX SamplePrep-Carver manual presses, the 25-ton 3628 Air-Actuated Bench-Press, and the 35-ton 3635 Automated X-Press. Pellet dies are available in the standard sizes of 13 mm, 31 mm, 35 mm, and 40 mm. Accessories include: aluminum Spec-Caps to form and protect 31 mm, 35 mm, 40 mm pellets; cellulose and paraffin Prep-Aid binders, as well as SPEX SamplePrep’s own UltraBind; and Sleeve-and-Plunger sets to form sample pellets with a thin layer of sample on a binder matrix.

While methods and mills for pulverizing are as varied as the samples themselves, pellet pressing requires simpler tools. SPEX SamplePrep presses, dies, and pelletizing accessories are sophisticated in design, but functionally simple and very effective.

Reproducibility is a cardinal virtue in sample preparation. SPEX SamplePrep dies and presses ensure production of uniform sample disks, whatever the sample or analytical technique.

Preparing Powder Samples for XRF

X-ray fluorescence spectrometry often requires the sample to be in a homogeneous powdered form with a planar surface. Although this can be accomplished by spreading the powder on the window film of a cell designed for liquid samples (such as a SPEX SamplePrep X-Cell), compression of the powder in a pellet die yields a denser, flatter surface that provides greater analytical accuracy and sensitivity, especially for wavelength XRF.

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In practice, the limiting par ticle size is often determined empirically by grinding the sample for a given length of time, measuring the particle size, analyzing the sample, then repeating at longer grind ing times until the intensities reach a plateau. SPEX SamplePrep offers laboratory mills and grinding vials that are ideal for this pro cedure.

Pressing Sample Disks

To make a sample disk, you need a sufficient quantity of homogeneous powder, often a mixture of thoroughly pulverized sample material and an appropriate binder; a pellet die set, including two pellets of polished steel or other hard, smooth material for sand wiching the sample powder; and a machine for exerting the necessary pressure on the powder. An optional aluminum cup such as the Spec-Cap contains the powder, protects the die bore against abrasive samples during pressing, and subsequently protects the pellet against chipping or breaking. An alternate technique, which yields a durable disk of solid binder with a thin layer of sample on one side, involves the use of a sleeve-and-plunger set with a pellet die.

The powder to be pressed must be fine-grained and homogeneous, in any case. If the pulverized sample material is not readily compressed into a stable, cohesive disk, it may be necessary to add a binding/briquetting agent such as cellulose, paraffin, or UltraBind. A binder can be uniformly blended with the sample or used as a backing for a thin layer of sample. Binders when properly used enhance analytical accuracy by stabilizing the sample disk and keeping it from distorting or crumbling during analysis and/or storage.

In an ideal sample disk pressed for XRF, the analytical surface of the disk is not only flat but also uniform, and truly representative of the original sample. This is true whether the sample was pressed without binder in a Spec-Cap, blended with a binder before pressing, or pressed in a thin layer on a binder matrix. Compact, flat, stable, and uniform sample disks can be ensured by the use of reliable SPEX SamplePrep dies, presses, accessories, and PrepAid binders.

While the pressing of powders into disks for XRF analysis is, in all its details, a fairly intricate exercise, it can be routinized to yield consistent, reliable analytical results. A review follows of the processes and tools involved in the actual pressing (pellet dies, binders, and anti-sticking agents, etc.). The discussion assumes that the analyte has been properly sampled (so that it is representative) and adequately pulverized (to minimize particle size effects).

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analysts do not bother to evacuate their 31 mm, 35 mm or 40 mm dies during pressing, but in fact the withdrawal of air and moisture from the sample can improve disk compaction and quality. Troublesome samples will often benefit from this technique. When evacuating a die, it is advisable to make sure both the upper and lower O-rings are in place and in good condition.

Set-Up and Loading of the SPEX SamplePrep Evacuable Pellet Die

Most evacuable pellet dies operate in the same way, by pressing the analytical sample between two polished pellets of steel or tungsten carbide. The simplest way of loading the die is to assemble the die barrel and die base; insert the lower polished pellet, polished side up, into the bore; pour the sample into the bore, level it, and add the upper polished pellet, polished side down; then insert the plunger, and place the assembled die in the press. This approach works well if the sample coheres well under pressure and is neither abrasive nor adhesive. Once the sample disk is pressed, it should be fairly easy to remove it from the die, either by hand or with the use of the knock-out ring supplied with each SPEX SamplePrep die.

When the sample does not hold together well after pressing, or sticks to the die or scratches it, there are various pelletizing aids available. These include Spec-Caps, PrepAid binders, Sleeve-and-Plunger sets, and various anti-sticking agents. Each has its proper application and technique, detailed later in this Chapter.

Care and Maintenance of the SPEX SamplePrep Evacuable Pellet Die

The SPEX SamplePrep evacuable pellet die is a precision tool which must be handled carefully and diligently maintained for proper operation. Although in design and function such dies are very simple, precise fit of the working parts is absolutely necessary, and easily jeopardized. A pellet die in good condition will produce thousands of sample disks without difficulty; a damaged or heavily worn die is likely to produce frustration, delays, and chipped or broken sample disks. Any damage to the polished pellets or the bore of a pellet die should be corrected immediately.

In a pellet die in proper condition, the polished pellets should pass smoothly through the die bore without binding, but their fit should be so precise that the pulverized sample will not “leak” around the pellet edges. A good test is to assemble the die bore and base, seal the evacuation port with a fingertip, and place a polished pellet (polished side up) in the bore. It should remain at or near the top of the bore, and spring back when pushed down lightly, due to compression of the air inside the die. When the evacuation port is unsealed, the pellet should drop smoothly to the bottom of the bore. If the polished pellet sinks immediately with the evacuation port sealed, it is either a loose fit, or the seal between the die body and base is damaged.

Polished pellets are made with close tolerances and sharp edges so that a sample powder will not “feather” into the gap between the pellet and the bore. (When this happens, the sample disk may have a raised, crumbly lip and the disk and upper polished pellet may be difficult to remove from the die). The edges of these pellets are by far the most vulnerable part of a die, as they can be dented or chipped by dropping them even a short distance onto a hard surface. Such dents are extremely dangerous to

Technique for Pressing a Sample Disk

In pressing samples for XRF, the loaded pellet die is placed in a hydraulic press, and the pressure is raised to a level that will cause the sample or sample/binder mixture to cohere into a stable sample disk. Manual and motorized hydraulic presses are available for this task: SPEX SamplePrep offers 12-ton and 25-ton SPEX SamplePrep-Carver manual presses, the 25-ton 3628 Air-Actuated Bench-Press, and the 35-ton 3635 Automated X-Press. Any SPEX SamplePrep pellet die will fit these presses.

A basic pressing sequence consists of raising the pressure to a specific level, holding it there for a certain length of time, and then releasing it, preferably slowly.

The maximum pressure for a given task varies considerably, depending on the size of the die and the nature of the sample. Obviously maximum pressures should not exceed the load limits of the die. The 13 mm die has a 10-ton limit, so some care must be exercised, but 31 mm and larger dies usually have load limits higher than either the capacity of the press or the pressure required to form a sample disk. Typical pressures for a 31 mm disk are from 20 to 25 tons; for a 35 mm disk, from 22 to 30 tons; and for a 40 mm disk, from 25 to 35 tons. Some samples cohere adequately at low pressures, but uniform high pressure is recommended. As “infinite depth” is very shallow for XRF analysis of most elements, a matter of microns, compaction of the sample decreases pore space and increases analytical accuracy.

Holding time and bleed time are both important. If a sample is simply brought to maximum pressure, and the pressure is abruptly released, the sample disk often does not hold together. This may be due to elastic rebound of gases trapped in the sample, because a binder may take time to completely penetrate the sample, or for other reasons. A holding time at maximum pressure of at least 30 seconds is recommended. Some analysts hold pressure for 5 minutes or more. During holding time the pressure should be maintained as well as possible. An advantage of the 3635 Automated X-Press is that it turns on the pump if the pressure drops more than 1 ton during the holding period.

A gradual release of pressure after the hold period is perhaps even more important than prolonged holding time. A minimum bleed time of 15 seconds is recommended. For samples that do not bind well, several minutes may be appropriate. A slow, careful bleed period can be difficult to accomplish with manual presses, as the pressure release control is often not sensitive, but it is still never a good idea to dump pressure abruptly in any pelletizing procedure. Another significant advantage of the 3635 X-Press is that lengthy hold times and precisely controlled pressure release can be programmed in, and will remain the same, sample after sample. The overall uniformity of the sample disks (and hence of the analytical results) will inevitably be greater.

The Use of the Vacuum in Pellet Pressing

All SPEX SamplePrep evacuable pellet dies have a vacuum hose attachment, which enables a vacuum pump to be hooked up to the die before and during a pressing operation. Most samples contain gases, moisture, and pore space, and removal or reduction of these can in fact affect the stability and uniformity of the sample disk. In the case of 13 mm dies, which are primarily used to produce KBr disks for infrared spectrometry, the use of a vacuum is necessary to draw moisture out of the KBr. Most XRF

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3615, 3617, and 3619 Spec-Caps acts as a lubricant, making it easier to remove the sample disk from the die.

There are two types of Spec-Caps, unpainted with flared walls (3619A, makes 31 mm disks) and painted with straight walls (3619, 30 mm, makes 31 mm disks; 3615, 33 mm, makes 35 mm disks; and 3617, 38 mm, makes 40 mm disks). The painted, straight-walled Spec-Caps can be used in two ways. The simpler is to fill the Spec-Cap with sample, level it off, and assemble the die around the filled Spec-Cap. This approach allows many samples to be set up and marked in advance, and when successful is quite efficient, but there are handicaps; perhaps the greatest is that unless the sample material is unusually dense or incompressible, the sample disk will be very thin, prone to cracking and possibly (in the case of low-Z elements) less than “infinitely thick.” There is also a chance of the Spec-Cap wall crumpling inward, and the technique of assembling the die around a loaded Spec-Cap requires some finesse.

Alternatively, the flared Spec-Cap technique may be used with either the painted Spec-Caps or the 3619A Pre-Flared Spec-Caps. Here the Spec-Cap is flared before being placed in the die; 3619 and 3617 Spec-Caps are flared by the user with the 3618 Edge-Flaring Tool, 3615 Spec-Caps are flared by the user with the 3625 Edge-Flaring Tool, and the 3619A Spec-Cap is flared at the factory. With this technique the die bore and base are assembled, the lower polished pellet inserted, and the flared Spec-Cap pushed down against that lower pellet. Then the sample is poured into the die, with the upper polished pellet and the plunger following. During pressing, the sample powder is forced inside the Spec-Cap. When the proper amount of sample is added, the top of the sample disk and the edge of the Spec-Cap will coincide. Obviously the sample weight will vary with the density of the sample and the size of the disk, but the general range of sample weight is, for a 31 mm disk, 5 to 8 grams; for a 35 mm sample disk, 7 to 10 grams; and, for a 40 mm disk, 8 to 12 grams.

Binders

Many analytical samples cohere well under pressure; those samples which crumble or ablate after pressing require a binder. Binders are usually blended with the sample after pulverizing and before pressing, but can also serve as a pellet matrix, supporting a thin layer of sample.

Binders can be liquids or powders, and range from commonly available reagents to brand name products with secret formulas. Their use should lead to a stable, crumble-proof sample disk achieved with a minimum of dilution, contamination, and effort.

Generic binders include cellulose, paraffin, boric acid, and graphite. Of these, cellulose and paraffin are available through SPEX SamplePrep in extra-pure, finely powdered form, ideal for blending and pelletizing. PrepAid® Cellulose (3642) has a particle size of less than 20 microns, and blends quickly and completely with samples at 10% by weight. It can also be used undiluted as a sample matrix with the sleeve-and-plunger technique. It will neither stick to a die nor contaminate a sample, and does not cake in the bottle during storage. Disks stabilized with cellulose should, however, be kept in a desiccator if they are to be retained, as with time they can absorb moisture and slowly swell and crack.

UltraBind, an exclusive SPEX SamplePrep product, satisfies our notions of the perfect all-around binder:

the integrity of the die. Not only can they cause the pellets to bind in the die, but also, under the high pressures of pelletizing, the stressed edge of the pellet can spall off. The resulting chip can be dragged through the bore, scarring it deeply, and potentially jamming the plunger and ruining the die.

Minor damage to the polished pellets and die bore should be immediately corrected with a fine-grained (e.g. 600 grit) emery paper. If after this the pellet will still pass smoothly through the die bore, and its leading edge is not significantly chipped, the pellet and die can continue in service. (A badly chipped pellet should be retired, as sample can wedge into the space left by the chip, making it difficult to extract the pellet from the die. In addition, further chipping is likely to occur under pressure). A lightly scarred die bore, properly smoothed, can continue in use.

Damage to the polished face of the pellet should also be avoided, but will probably not affect the functioning of the die. Analytical accuracy is what suffers. Scratches on the order of 20–30 microns can cause shielding effects in the sample disk, and overall abrasion of the polished pellet face can very slightly change the geometry and distance in the critical relationship between X-ray tube, sample disk, and detector. Obviously if analytical results are being distorted because of the condition of the polished pellets, it is time to replace them, but the degree to which such distortion is tolerable will vary considerably from user to user. A simple way to check polished pellets is to press two sample disks of identical material, one with a pristine polished pellet and the other with the worn pellet, and compare the analytical data.

In handling the pellet die, some simple rules should be kept in mind: keep the die clean, and always treat it as the precision tool it is. Pellet dies should be cleaned after every use, to avoid both sample cross-contamination and the possibility of disk jamming or sticking from sample build-up. In cleaning the polished pellets, treat them like glass; in other words, use the same cleaning technique you would for a glass lens or mirror. Steel has a hardness similar to glass, and it is important to avoid scratching the polished surface.

Remember that the polished pellets are the most critical parts of the die, and the most easily damaged. When inserting the polished pellets into the bore of the die, take extra care that they do not jam; the fit is so precise that a very slight tilt will cause them to stick. When this happens, free the pellet gently. Above all, do not push it down further and make the situation worse, as this can cause the pellet to chip, and quite possibly ruin the die. A simple technique for inserting the polished pellet into the die bore is to hold the trailing edge of the pellet lightly with the finger-tips, and rotate it gently in the mouth of the bore to make sure it is properly lined up. When a polished pellet is placed in the diebore, it should move freely. If it does not, careful corrective action should be taken immediately.

Proper Use of the SPEX SamplePrep Spec-Cap

Spec-Caps are shallow aluminum cups in which a sample is pressed. A sample disk properly prepared with a Spec-Cap will be encased by the Spec-Cap on one side and around its edge, allowing the disk to be written on and handled without crumbling or contamination. The Spec-Cap also protects the bore of the die from abrasive samples, which with time can enlarge or damage the die, and the paint on the

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to attain very fine particle size. When a grinding aid such as Vertrel® XF (3650) is used, the binder can usually be added to the sample before grinding.

Sleeve-and-Plunger Technique: Binder as Sample Disk Matrix

Besides being blended with a powdered sample, binders can also be used in unblended form as a sample support. A typical XRF disk made with a SPEX SamplePrep Sleeve-and-Plunger Set consists of a thin layer of powder supported by a binder matrix. Since most of the X-ray fluorescence emission measured by an XRF spectrometer comes from within a few microns of the surface, the technique is successful with small quantities of sample, often a gram or less. Other advantages are that the disk is quite strong and stable without a Spec-Cap, is easy to remove from the die, and will not scratch the bore of the die. The sample is also presented to the spectrometer in nearly undiluted form, as the binder penetrates the pore space between particles rather than separating them. (Binders have a much lower specific gravity than most sample materials. Even though samples are typically blended with 10% binder by weight, the disk may have a much higher proportion of binder by volume, diluting the sample and diminishing its XRF emissions).

When a SPEX SamplePrep Sleeve-and-Plunger Set is used to make XRF sample disks, this is the technique. First the barrel and base are assembled, and the lower polished pellet placed in the bore of the die. Then the sleeve is inserted in the bore. The sample is poured onto that part of the polished pellet left exposed, and the plunger inserted in the sleeve to even out the sample powder and tamp it down. Now the plunger is carefully withdrawn followed by the sleeve. This leaves a thin layer of formed sample in the die, onto and around which the binder is poured. Finally the upper polished pellet is inserted in the bore, followed by the die plunger, and the assembled die is transferred to the press and put through a pressing cycle. The finished disk consists of solid binder with a uniform sample layer “bull’s-eye” in the target area of the disk. Sleeve-and-Plunger Sets are available for SPEX SamplePrep 31 mm, 35 mm, and 40 mm dies, and are described, beginning in Section 1, Chapter 5 of this Handbook.

Sample Disk Removal

When the sample disk has been formed, and the die taken out of the press, the disk must then be removed from the die. In some cases this can be done by inverting the die and standing it on the plunger, removing the base, and pressing down on the edge of the die barrel with the heels of the hands. As the die barrel moves down, the plunger pushes the polished pellets and sample disk up. If hand pressure will not budge the plunger, the knock-out ring (supplied with each SPEX SamplePrep pellet die) should be used.

The knock-out ring is a tube which fits into the recess in the lower end of the die barrel. With the die base removed and the knock-out ring in place, mechanical force can be applied to the die, easing the polished pellets and the sample disk out of the die without damaging them.

it blends well with samples, and is self-lubricating, strong when palletized, and moisture-resistant. Most batches, however, have low amounts of NaCl as a relic of the manufacturing process. A fine (20 µm) powder, Ultrabind blends easily with samples to yield a disk which is easy to remove from the die, and durable enough to withstand rough handling. It also resists cracking and swelling in storage, and is an excellent disk matrix when used with the Sleeve-and-Plunger technique, described in this chapter. UltraBind® (3644) is described on page 200 along with other PrepAid binders.

Sample disks formed with 10% to 20% paraffin are air-stable, as the waxy binder seals the surface against moisture. Powdered Paraffin (3646) is also offered by SPEX SamplePrep. As with cellulose, the fine (30 µm) powder blends evenly with samples and does not contaminate them. However, paraffin should not be used undiluted with the sleeve-and-plunger technique, as in this concentration it will stick to the die.

Boric acid is commonly available in pure form, and can be used diluted or undiluted to make a stable sample disk. However, it is much more hygroscopic than cellulose and hence it typically supplied in granular form, requiring grinding in a mill or mortar-and pestle to become a fine enough powder to blend evenly with a sample. Some analysts add boric acid to the sample during the final stage of grinding to achieve this. Sample disks bonded with boric acid must be kept in a desiccator if they are to be preserved.

Graphite powder makes sample disks unaffected by moisture, and blends rapidly with any sample. It also serves a its own lubricant, making it easy to free disks from the pellet die. Its disadvantage for XRF is that it must be used in a proportion of at least 50% of the sample disk by weight. Graphite was used extensively in the development of sample-disk pressing techniques, but that was because of its high conductivity. The 31 mm sample disk accepted as the standard size by most XRF manufacturers and users was originated for the point-to-plane technique in arc/spark optical emission spectroscopy, where a high-voltage current is passed through the sample. Sample disks pressed with graphite are also used for spark ablation techniques.

Solid binders also include powdered wax, pulverized acrylic plastic, and others. Liquid binders are also used; they are blended with a sample prior to pressing. Most appear to be a solution of polymer(s) in solvent(s); their reputed advantage is that during blending, all the sample particles receive a light coating of the polymer, so that subsequent pelletizing is more direct and efficient.

There are also materials which are touted as aids for both grinding and binding. Generally these are in tablet form, so that a uniform amount can be easily added to the sample just before grinding. The classic example of this genre is aspirin, but there are more sophisticated, proprietary grinding/binding tablets available. Of the use of binders in general, it should be remarked that their most conspicuous property is that of caking under pressure, or at least of flowing under pressure and then caking when the pressure is relieved. Hence there is a good chance that adding a binder to the sample before grinding will lead to caking in the grinding container. In this connection it should be noted that a very finely ground sample (e.g. cement-mix ground to below 10 microns) is more likely to bind well under pressure than the same sample ground to moderate fineness, and that grinding aids are often needed

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Sample Disk Handling and Care

For maximum precision in XRF analysis, the surface of the sample disk should be very flat and very clean. It follows that these disks should be handled carefully, to avoid both surface contamination and damage. Sample disks should be handled only by their edges, and preferably not with bare skin; sweat, skin oils, and incidental contaminants are easily transferred to the analytical surface of the sample disk.

Physical damage to the disk surface also affects accuracy: scratches, chips, pits, and swelling or curving should all be watched for and guarded against. As a general rule, it is best to analyze a sample disk as soon as possible after it is pressed. If it must be kept as a reference sample or standard, it should be preserved in a desiccator. Most pressed powders will absorb moisture with time, and swell. Some samples exhibit a kind of elastic rebound shortly after pressing. (Flatness can be checked by holding a sample disk against a polished pellet). Sample disks can also be degraded by X-rays. Even the heat produced by the X-ray beam can cause small flakes or particles to break loose from the face of the disk, leaving the disk with an irregular and possibly non-representative sample surface, and contaminating the spectrometer.

There can be sample disks which, after the use of binders and the most careful pressing procedures, will still degrade in the spectrometer. The most common remedy is to place a piece of XRF window film (e.g. SPEX SamplePrep 4 micron Ultralene) where it will protect the spectrometer; one ingenious solution has been to place a pressed 31 mm sample disk inside a SPEX SamplePrep 3527 40 mm X-Cell.

If a pressed sample disk is to be used as a standard, it is advisable to have that standard made in duplicate, with the spare disk carefully preserved. The standard in daily use should be periodically compared with the standard kept in reserve. Often the first symptom of sample disk degradation is a decline in analytical accuracy.

Whether the sample disk is extracted by hand, or with the press and knockout ring, the die barrel and plunger should be kept in an inverted position to minimize the risk of damaging the sample disk during extraction.

The use of lubricants in the die usually makes it possible to extract the sample disk by hand, as well as minimizing the chances of the sample disk jamming or sticking in the die bore. Consult the Anti-Sticking Agents section, detailed below, for additional information.

Anti-Sticking Agents

In commercial molding operations, whether of plastics, metals, or other materials, releasing agents are often necessary to effect the easy removal of the product from the mold. This is the case with XRF sample disk preparation, but releasing agents are not widely used because of concerns about sample contamination. Even samples which are not inherently sticky can become somewhat adhesive when forced against a steel plate by 25 tons of ram pressure, and in general it is difficult to remove a pressed sample disk from a die by hand pressure alone.

The use of the press together with the die’s knock-out ring is common practice, and as long as the disk does not bind to the die bore or the polished pellets, the knock-out ring approach works well enough. However, the use of a lubricant in the bore of the die does not have to contaminate the face of the sample if that lubricant is carefully applied, and the sample disk is carefully handled during and after removal from the die. For example, there is a fluoropolymer spray which can be squirted into the bore of a die before the die is loaded. After application, the volatile carrier evaporates, leaving a thin coating of fluoropolymer on the die bore. When the die is then loaded and the sample pressed, the sample disk and the polished pellets should have lubricant only along their edges, and should be easy to remove from the die. Even so, any such lubricant should be thoroughly tested before it is incorporated in a pressing pro cedure.

Every XRF analyst knows the anguish of having a sample disk emerge from the die firmly attached to the polished pellet; at best the disk is ruined. This may be due to the pellet being damaged or poorly cleaned, but it is more likely that the sample is at fault. While spraying the polished pellet with lubricant will prevent sample adhesion, it will also contaminate the face of the sample disk with lubricant. The simplest non contaminating approach is to place a piece of thin plastic film between the polished pellet and the sample before pressing. SPEX SamplePrep 3516 Mylar window film is recommended because of its 0.12 mil thinness. After the sample powder is added to the die and leveled, a piece of film can be placed over the bore, and the upper polished pellet pushed down on top of it. (A more elegant approach is to cut circles of film the size of the die bore. When the pressed sample disk has been removed from the die, the film is peeled off and discarded).

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To prevent further damage, always remember. . . Never drop the pellets, or allow the edges to become dented. Never tilt the pellets so that they become jammed when being placed in the die. (If they become jammed, free them as gently as possible). Always put the squared-off end of the plunger into the die; if the stepped/beveled end of the plunger contacts a polished pellet under pressure, the pellet can chip.

Selecting a Pellet Die

The choice of die is generally determined by the requirements of the analytical instrument. SPEX SamplePrep offers dies to make four standard sizes of sample disks: 13 mm for IR spectrometers, 31 mm for most OES and XRF instruments, and 35 mm and 40 mm for newer XRF spectrometers. Dies are clearly labeled with bore size and maximum safe load.

Each SPEX SamplePrep die set is a complete unit. Made of hardened stainless steel for durability and extra wear, the SPEX SamplePrep die includes a body with detachable base, a plunger, and two polished steel pellets. Sample material is pressed in the die bore between the polished pellets, yielding a compact sample disk ready for the spectrometer’s sample holder. A convenient “knock-out ring” allows easy extraction of the steel pellets and sample disk from the die. Each precision-machined SPEX SamplePrep die set also incorporates a vacuum hose attachment. This allows evacuation of gases, volatiles, and moisture during pressing, assisting compaction and preventing possible sample disk rupture under vacuum-path conditions.

Trouble Shooting

A carefully conceived and implemented disk pressing procedure should give the analyst few difficulties. However, it is flying in the face of experience to assume that nothing can go wrong. The following pointers are essentially an index to this section of the Handbook, indexed by problem:

If the sample disk comes out of the die smoothly, but breaks. . .

• increase pressure, hold time, and/or bleed time .• use Spec-Caps.• use a binder.• grind the sample finer.• evacuate the die during pressing.

If the sample disk is hard to remove from the die. . .

• use Spec-Caps.• use a lubricant.• prepare disks with a binder matrix.• check that the bore of the die and the polished pellets are clean and undamaged;

maintain or repair as necessary.

If the sample disk sticks to the polished pellet. . .

• put a thin sheet of SPEX SamplePrep Mylar, polypropylene, etc. between the sample and the polished pellet, before pressing.

If sample disks degrade rapidly in use. . .

• use a binder.• evacuate the die during pressing.• protect the spectrometer with SPEX SamplePrep XRF Window Film.• store the sample disk in a desiccator if it is to be retained.

If the polished steel pellets become scratched in normal usage. . .

• grind the sample finer.• use polished tungsten carbide pellets, e.g. SPEX SamplePrep 3623C.

If the polished pellets become chipped or jammed. . .

• STOP IMMEDIATELY, and take corrective action to avoid making matters worse.

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Most of these samples are naturally oxygen–rich and do not require chemical transformation prior to borate fusions. However, hybrid oxidation/fusion techniques have been developed for reliable borate fusions of sulfides, carbides and some ferro-alloy materials formerly considered out-of-bounds for the technique.

Borate fusion has become increasingly popular as a preparation technique for XRF sample disks. Because fusion eliminates particle size and mineralogical effects and produces a homogeneous sample, it has proven to be the best method for materials that have these characteristics when X-ray fluorescence analysis is the method of choice. The fusion method will reduce matrix effects but not eliminate them. Borate fusion may not be the method of choice if the analyst is interested in trace analysis, since the sample is diluted during fusion. There have been recent advances in improving the performance of fusions for trace analysis.

The pressed powder method can be highly accurate when carefully done with multi-phase samples such as cements, rock, sand and ore but such samples are subject to segregation during grinding and pressing, and to matrix effects (e.g. particle size and mineralogical effects). Borate glass disks are easier to preserve than pressed powder disks because they are stable if carefully stored in desiccators. Synthetic standards for XRF can also be made from pure oxides with the borate fusion method, as borate glass is essentially a solid solution with few matrix-matching problems.

In preparing samples for AA, ICP, and other liquid-analyzing techniques, the major advantage of borate fusion is that it is often simpler and quicker than dissolution with acid in a microwave pressure vessel. A complete fusion/solution procedure, from ignition of the heating elements to decanting of a clear solution, can take fifteen minutes or less in an automated fluxer. While borate fusions do require some caution in evacuating heat and fumes, and the use of dilute HCl or HNO3 to dissolve the melt, hazardous reagents such as HF and other concentrated mineral acids are not necessary.

Borate fusion methods offer a wide range of applications but may not be suitable for all materials. Fusion destroys the original form of the sample, so structural and molecular information should be measured before the fusion is made. The high temperature of borate fusion (1000˚ to 1150˚ C) drives off compounds of volatile metals such as Hg, Sn, and Sb, while other compounds form during fusion. Extra steps necessary to prepare organic materials and reduced inorganics for fusion can extend turnaround time but still may be the most accurate method to choose. For many samples borate fusion is the simplest, quickest, and most accurate analytical approach.

Both fusion and pulverizing/pressing are important and widely used sample preparation methods, each with their own advantages. SPEX SamplePrep has a full range of equipment for either approach. Please consult our applications specialists to help determine which method is more suitable for your laboratory.

Introduction

Borate fusion is an extremely effective method of preparing cement, refractories, ceramics, rock, and similar materials for elemental analysis by XRF, AA, and ICP. The samples are mixed in powdered form with a flux, either lithium tetraborate or a mixture of lithium tetraborate and lithium metaborate. The sample must be ground to a fine powder before the fusion; the more consistent the particle size, the more reproducible and accurate the fusion will be. The sample mixture is heated until the flux melts and the sample dissolves in it, yielding a homogenous melt. The sample forms borate salts and this eliminates particle size and mineralogical effects. The use of borate fusions will also minimize matrix effects which are seen in XRF analysis. The melt can be cast as a glass disk for XRF, or quickly dissolved in dilute nitric acid or hydrochloric acid for analysis in solution by AA or ICP. Recent advances also allow borate fusion of samples containing sulfides, ferroalloys, etc.

One of the advantages of borate fusion is the short preparation time, typically ten to fifteen minutes to make glass disks or solutions with automated fluxers. For samples prepared as liquids, borate fusion can be quicker overall than microwave dissolution in pressure vessels, and the use of hazardous acids (e.g. HF) is avoided.

SPEX SamplePrep offers two approaches to borate fusion. The Katanax K1 and K2 Prime Series Automated Electric Fluxers for rapid, reproducible fusions, and graphite crucibles for smaller-scale operations with muffle furnaces. SPEX SamplePrep also provides the full line of SPEX Fusion Flux.

When to Employ Borate Fusion

Borate fusions are widely used for samples which are either difficult to prepare as homogenous pressed powder (e.g. cement), hard to dissolve in acid (e.g. zirconia and alumina), or both (e.g. metal ores and silicate rocks). For borate fusions to be successful, the sample when fused must be in the form of an oxidized, inorganic compound. Cement is usually a blend of carbonates and silicates; zirconia and alumina are oxides; and so forth. Compounds without oxygen such as sulfides, carbides, chlorides, etc, must be oxidized before being fused. Reduced metals must also be oxidized. Organic compounds must be ashed. (An example of this is the analysis of metallurgical coal. The coal sample must be ashed and the ash is then fused. The sulfur in the coal will volatilize in the process and therefore sulfur cannot be measured). Once fusion is complete, the melt can be cast as a glass disk or poured into a dilute acid solution and dissolved. Platinum-group metals cannot be fused with borates because these compounds reduce during fusion and the metals will not only remain insoluble in the flux but can also alloy with the 95% platinum/5% gold crucibles and damage or destroy them.

Many users of the K1 and K2 Prime Series Automated Electric Fluxers are in the following industries: cement, glass, ceramics, mining and minerals. Samples analyzed include not only raw materials like dolomite, sand, basalt and iron ore, but also their industrial products and by-products such as cement, building materials and mining concentrates. Additional industrial samples include pigments such as TiO2, and slags from smelters, blast furnaces, refineries and glass plants.

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Other fluxes include sodium tetraborate (Na2B4O7), sodium metaphosphate (NaPO3), and potassium pyrosulfate (K2S2O7). These have lower melting points than lithium borate fluxes and more specialized applications. Alternative fluxes and fluxing techniques are discussed in Bock (1979) and in Sulcek and Povondra (1989), both cited in the bibliography at the back of this Handbook. Melting point may be a factor in the selection of a flux, as the higher temperature of a fusion, the greater the degree of volatilization. However the utility of lithium tetraborate and lithium tetraborate/metaborate mixtures is so great that most analytical fusions are carried out with these fluxes at temperatures between 1000˚ and 1150˚ C.

SPEX SamplePrep offers the full line of SPEX Fusion Flux. This includes lithium tetraborate, lithium metaborate, several lithium tetraborate/metaborate blends, sodium tetraborate, and a lithium tetraborate/lithium carbonate mixture. Many of these fluxes are available in pure and ultra-pure grades. All are supplied with a certificate of analysis for trace metal impurities.

The most popular blended flux, 2:1 “tet:met,” is also available with added LiBr non-wetting agent for greater convenience. LiBr solution is available separately for quick addition of a non-wetting agent to a flux.

Most SPEX Fusion Flux are of the “glass bead” type, pre-fused and coarse-grained for maximum homogeneity and ease of use, and minimum absorption of moisture. The exceptions are the lithium borate/carbonate flux (a physical mixture) and so-called “fluffy” lithium tetraborate. The latter is fine-grained and less dense than the “glass bead” fluxes, but is available for those who find its physical characteristics superior for their application.

All fusion fluxes sold by SPEX SamplePrep are manufactured to the highest standards available. SPEX SamplePrep guarantees both the purity and the quality of the fluxes and additives supplied to our customers.

Additives for Fusions: Non-Wetting Agents, Fluidizers, and Oxidizers

Non-wetting agents (NWA) are iodides and bromides which can be added in small quantities to a fusion so the molten flux will not stick to the crucible or mold. The visible effect of a non-wetting agent is to increase the surface tension of the melt. A fused disk with too little NWA will have a concave upper surface and may be difficult to remove from the mold, whereas a molten flux bead with excessive NWA will ball up when poured and not form a complete disk.

When glass disks for XRF are being made, a non-wetting agent is mixed with the flux and the sample before fusion starts. Typically the amount of NWA is about 0.2% of the weight of the flux, e.g. 12 mg of NWA for 6 grams of flux. Certain samples such as iron ores, which greatly increase the “stickiness” of a melt, require additional NWA. As non-wetting agents gradually volatize during a fusion, somewhat longer fusions (as for some technical ceramics) may also need greater amounts of NWA. The ideal amount of NWA for a specific procedure is usually determined by experiment.

How to Prepare a Fused Sample

Fusions are accomplished in several steps. First the sample is mixed with a flux in an appropriate ratio (usually between 1:2 and 1:10), with the addition of a non-wetting agent to prevent the flux from sticking to the crucible and the mold. Typical amounts of flux/sample mixture are 6 to 7 grams for a 31 mm glass disk, and 1 to 2 grams for a solution. The sample is heated past the melting point of the flux in an inert, heat-resistant crucible. Most borate fusions are performed in crucibles made of 95% platinum and 5% gold, a standard non-wetting alloy. Some borate fusions are done in graphite crucibles.

During the fusion, heating is maintained and the crucible regularly agitated until the sample has completely dissolved in the molten flux. At this point the melt is either poured into a mold and annealed to form a glass disk for XRF, or poured into dilute mineral acid (e.g. 10% HNO3) and stirred until the glass flux dissolves. In some cases (notably pyrosulfate fusions) the melt is left to harden in the crucible, and the crucible and the glass together are placed in an acid solution to dissolve the glass.

It is possible to achieve great sample throughput with an automated, programmable fluxer. The Katanax K1 Prime, when programmed for cement samples, can produce up to seven fused glass disks in an hour, using 0.6 grams of cement mixed with 6 grams of flux and about half a percent of LiBr, a non-wetting agent. The 5-position Katanax K2 Prime, when programmed for similar samples can produce up to 25 fused disks per hour.

The same borate fusion procedures can be carried out manually in a muffle furnace with SPEX SamplePrep graphite crucibles. The larger flat-bottomed SPEX SamplePrep crucibles can be used to cast glass disks as well as perform fusions, while the crucibles are handled and agitated with SPEX SamplePrep tongs. The time per sample with the muffle furnaces is usually much longer than with an automated fluxer.

Each approach has its advantages. Again, please contact our applications specialists to determine the optimum equipment for your requirements.

How to Select a Fusion Flux

Most fusions involve the use of lithium tetraborate (Li2B4O7, M.P. 920˚ C), lithium metaborate (LiBO4, M.P. 845˚ C), or a mixture of the two. As a rule lithium tetraborate is better suited for the dissolution of basic oxides, and is preferred for cement and most ores. Lithium metaborate or “met/tet” mixtures are more suitable for acidic oxides such as silicate rocks and silica-alumina refractories. Individually or together, these lithium borates will dissolve oxides, carbonates, silicates, sulfates, etc. Metals, sulfides, nitrates, carbides, phosphides, etc. cannot be fused directly in lithium borate fluxes, and will often attack platinum-gold crucibles, or alloy with them. However, many of these materials can be first oxidized with standard techniques and then successfully fused. Methods have been developed for fusing sulfide-rich material such as copper ore. The sample is mixed with lithium or sodium nitrate and preheated to oxidize the sulfide to sulfate. When this has been done the fusion can proceed as normal without any loss of sulfur from the fusion.

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Sec tion 2Fusion

SPEX SamplePrep flat-bottomed graphite crucibles (7157– 7162) are designed both as fusion crucibles and as molds for XRF cast glass disks; fusion of the sample and annealing of the disks can be completed in the same container. These crucibles have tapered walls to facilitate removal of the glass disk. The three different crucible sizes (31 mm, 33.6 mm, and 40 mm) produce disks which will fit the standard XRF masks of 31 mm, 35 mm, and 40 mm. The crucible bottoms are flat but not polished, so that for greatest analytical accuracy the disks should be flat-lapped.

While graphite is an ideal crucible material for many applications, withstanding temperatures well in excess of any required for even the most difficult borate fusions, it does oxidize slowly above 430° C; over a period of hours some erosion of the crucible can occur, the graphite converting to CO2. Because of this, graphite is not recommended for extremely lengthy fusions, or for fusions where the sample might be reduced.

SPEX SamplePrep also offers a complete line of borate fusion fluxes and non-wetting agents.

When making solutions by pouring the molten flux into a dilute mineral acid, it is desired to have complete transfer from the crucible to the beaker. This can require a much higher proportion of non-wetting agent than is necessary to pour a glass disk. A quantity of flux plus sample not exceeding 2 grams might require 50 to 100 mg of NWA. Lithium iodide and bromide are popular non-wetting agents because they do not add an impurity to the flux. However lithium bromide is hydroscopic, so it is usually made into a saturated solution and added to the flux from a dropper bottle. Lithium iodide, sodium iodide, and cesium iodide are somewhat more air-stable, and easier to use as solids. While it is simpler to add a drop or two of NWA than it is to weigh out 10 or 20 mg of a solid, liquid NWA cannot be added to a hot crucible while a fusion is in progress. Note that non-wetting agents should be used with care when copper-bearing samples are being fused, as copper halides are extremely volatile.

Lithium fluoride can be used as a fluidizer, lowering the melting point of a flux and making it flow far more easily. At 10% by weight, it lowers a flux’s melting point by about 100˚ C.

Oxidizers such as lithium nitrate and sodium nitrate can be useful in eliminating unoxidized components from a sample that will not fuse. Graphite, often present in cement mix, is relatively harmless but can leave a black film on a glass disk or even cause it to crack. Graphite can be oxidized to CO2. Other sample components such as phosphides and sulfides may be corrosive enough to damage or wreck a crucible in a single fusion. If they are oxidized to phosphates and sulfates they will be comparatively harmless, and their cations will be present in the fused glass disk for analysis. As oxidizers have much lower melting points then borate fluxes, any fusion including them should proceed at a low temperature until oxidation is complete.

Graphite Crucibles and Handling Equipment

SPEX SamplePrep graphite crucibles are a cost-effective alternative to the metal crucibles (platinum/gold, zirconium, etc.) used to contain samples during fusions. SPEX SamplePrep graphite crucibles are disposable, eliminating both the need for time consuming cleaning procedures and the possibility of cross-sample contamination. Chemically inert and heat-resistant, graphite will not combine with samples during fusion. And SPEX SamplePrep’s unique nickel-chromium rack and tongs make handling easy.

SPEX SamplePrep graphite crucibles are available in two purities: high-purity, for trace-level analytical work, and economical regular-purity, good for most fusions. SPEX SamplePrep high-purity graphite has total metallic impurity (TMI) limits of 10 ppm, measured by DC arc spectrometry. SPEX SamplePrep regular-purity graphite typically has from 10 to 100 ppm each of Mg, Fe, Al, Si, and Cu; as these impurities are distributed throughout the graphite, contamination of the sample is minimal.

SPEX SamplePrep 7152 is the standard 9 mL fusion crucible found in most laboratories; long-time users should note the introduction of the 7152HP, the same crucible in purified graphite. For fusions where the 7152 is too small, SPEX SamplePrep 7156 is a standard graphite crucible with about three times the capacity of the 7152. (7155 is the high-purity version of the 7156).

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polyethylene, closed X-Cells are avail able in both 31 mm and 40 mm sizes to fit most spectrometers. For special applications there are 31 mm Double Open End and Micro X-Cells, and a 43 mm Closed Cell for Horiba Sulfur Analyzers.

The patented closed X-Cell design features a snap-open seal for vent ing volatile samples and a reservoir cup for catching any liquids that expand through the vent. This assures the analyst of a smooth, flat window for improved precision, yet guards against spillage in inverted-optic spectrometers. A roughened surface within the reservoir cup facilitates labeling.

Each closed X-Cell consists of a cup and snap-on ring. For routine liquid samples, the cup is filled with liquid, a piece of film is spread over the top, then the snap-ring is pushed down over the cup to form a flat drum-like window. The cell is turned over and, if desired, the snap-post on the rear can be pushed to break the seal and to permit venting of trapped bubbles or expanded liquid while the sample is run. The filled X-Cell is then positioned in the spectrometer mask.

X-Cells can also be used to run solution residues and small quantities of pastes or powders. For solution residues, the film window is prepared as for solutions, then dished slightly with the rounded end of a glass rod. A drop of solution placed there and warmed under an IR lamp dries to a smudge for in situ analysis. If powder or paste is spread on the surface and another piece of film overlaid before snapping on the ring, the sandwiched sample is ready for X-ray analysis. The 3527 40 mm X-Cell can also be used to hold crumbling 31 mm sample disks.

SPEX SamplePrep also supplies versatile Double Open End X-Cells which are ideal for many unusual or hard-to-handle samples: slurries, sludges, pastes, viscous materials such as glue, tar, or RTV sealant, metal films and machine parts, etc. The 3571 X-Cell has a body open at both ends, two snap-on rings, and a collar to hold film in place while the ring is being applied. After window film has been attached to one end of the cell, it can be inverted, and the sample placed, poured, dabbed, or otherwise located in the cell, directly against the window. The top of the 3571 X-Cell can then be left open, sealed with film, or protected but allowed to “breathe” with a piece of 3524 Microporous Film.

If you deal with small air-sensitive or hazardous samples, then the 3577 Micro X-Cell is the solution to your problems. While it fits in the standard 31 mm sample holder, the window is 6.3 mm in diameter, and only 0.5 mL fills it up. Collimate your beam down to reduce scatter and you’re ready to go. The Micro X-Cell can be run with an open or window back, but it will also accept a 7 x 13 mm serum bottle closure for injection, purging, or the isolation of special samples.

Bubble-Free Cell Insert

The Bubble-Free Cell Insert is a simple, patented insert for the 40mm XRF liquid cell (3527) which prevents trapped air from forming a bubble under the window film when the cell is used in the upright position. The 3527I Bubble-Free Cell Insert is pushed into a cell before the sample is added, and forms an inverted-cone shelf under which air bubbles can be trapped once the cell is filled and sealed.

In many XRF systems, liquid cells are run in an inverted position, so any bubbles rise to the back of the cell and do not interfere with the analysis. The drawback of this “inverted optics” configuration

Introduction

The XRF spectroscopist is confronted with an almost infinite variety of samples. What they have in common is that they must all be presented to the XRF spectrometer in the form of a homogeneous, flat disk. Liquid samples, from water to oil, are run in sample cells covered with a thin film. Many solid samples, if homogeneous, can be machined to the proper shape and run directly. Others, such as refractories, ores, and biological materials, are too inhomogeneous and must be pulverized or fused. Then the homogenized material can be dissolved for analysis as a liquid sample, pressed into pellet form, or cast as a glasslike solid.

SPEX SamplePrep manufactures disposable sample-handling accessories for powder and liquid XRF samples. Over the last twenty years, millions of powder samples for XRF have been pressed and protected in SPEX SamplePrep Spec-Caps. Millions more liquid XRF samples have been run in SPEX SamplePrep X-Cells. These and the related products listed here will save you time and money, and assure you of reproducible, contamination-free results.

SPEX SamplePrep also provides an extensive line of sample preparation equipment particularly suited for XRF spectroscopy. There are laboratory mills and grinding containers capable of converting most solid samples into homogeneous powders, as well as dies and presses capable of compressing those powders into flat, uniform sample disks, ready for XRF analysis. There are also the Katanax K1 and K2 Prime Automated Electric Fluxers for rapid, automated borate fusions, yielding XRF glass disks, or solutions for AA, ICP, etc. For further fusion work there are also full lines of graphite crucibles, borate fusion fluxes, and fusion additives such as non-wetting agents and oxidants. These products are described elsewhere in this handbook; please feel free to consult our sample preparation specialists to determine the optimum equipment and techniques for your application.

Reinforcing XRF Pellets with Spec-Caps

SPEX SamplePrep Spec-Caps are shallow, thin-walled aluminum cups which are routinely used in the production of pressed-powder sample disks for XRF. The Spec-Cap forms the bottom and partial sides of the completed sample disk (also called pellet, briquette, or planchet). Thus reinforced, the disks are resistant to chipping and breaking and are more easily handled, marked and stored than unclad disks.

The sample holders of most X-ray spectrometers will accept 31 mm (1¼ in.) sample disks. For these instruments, 3619 and 3619A Spec-Caps are ideal. Several spectrometer manufacturers have shifted toward 35 mm and 40 mm sample disks, and for these larger sizes we recommend 3615 and 3617 Spec-Caps, respectively.

X-Cells for XRF Liquid Samples

SPEX SamplePrep X-Cells are used throughout the world for fast, dependable, reproducible running of liquid samples in XRF spectrometers. They are designed to fit into the major suppliers of XRF spectrometers such as PANalytical, Rigaku, Bruker, and Thermo Instruments. The requirements of the application will dictate the type of X-Cell and which window material to use. Made of contaminant-free

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2. Transmission: X-ray transmission through polymer films is affected by the thickness of the film as well as its composition. Of the SPEX SamplePrep XRF window films, 8 µm Kapton is the least transparent to X-rays; 6 µm Mylar is more transparent, as is 5 µm polypropylene. The thinnest films (4 µm Ultralene, and 3 µm Mylar) have the greatest degree of X-ray transmission, and should be considered for light-element analysis in particular.

3. Uniform Thickness: The absorption of X-rays at a given wavelength is directly proportional to the thickness of the film. Thus you will get uneven fluorescent intensity with an uneven film. SPEX SamplePrep films are selected for uniform thickness so your results are reproducible.

4. Purity: No XRF window film is absolutely free of metallic impurities, but some are cleaner than others. While Mylar film makes a suitable window for most analyses, it may contain trace levels (ppm) of Ca, P, Fe, Cu, Zn, or Sb. Polypropylene film has been known to contain trace levels of Ca, Zr, P, Fe, Zn, Cu, Ti, and Al. Our cleanest film overall is Ultralene. Kapton is clean in most respects but now includes a phosphate-based surface coating to improve its handling characteristics. Our best advice is that you run a blank from every new roll or box you open since impurities can be different not only between types of film but also between different lots of the same material. SPEX SamplePrep cannot be held responsible for variation in film composition.

5. Chemical Attack: Your sample’s chemical characteristics may dictate your choice of window film. Some samples simply attack some films, exposing the spectrometer to possible damage if the window leaks. For each new sample, we strongly recommend that you test the window film by exposure to the sample for several times your longest anticipated running time. The tables which follow may be of some help in selection, but they are not meant to replace actual testing.

In general, Kapton is highly resistant to acids and organic chemicals and almost everything except strong bases. Mylar is vulnerable to strong acids and strong bases and resistant to organic chemicals. Polypropylene and Ultralene are reasonably resistant to acids, bases, and many organic chemicals, but can be attacked by aromatic and halogenated hydrocarbons. All of these films are attacked by strong oxidizing agents. Microporous Teflon is not used as a window film as such, but is essentially impervious to chemical attack.

X-Cells are made of polyethylene, which is generally resistant to acids, bases, and most organic chemicals except halogenated hydrocarbons. Window films, being much thinner than X-Cell bodies, are likely to perforate long before the X-Cell is affected. However, polyethylene begins softening at temperatures well below 100°C, so if you are working with hot samples, pre-test the X-Cell body as well as the window film.

CAUTION: SPEX SamplePrep window films and X-Cells are not intended to be used in vacuum-path XRF systems. An abrupt pressure change in an XRF spectrometer’s sample chamber can stretch or burst window film, or otherwise cause the X-Cell to leak or come apart. It should also be noted that some volatile chemicals have the ability to generate pressure inside an X-Cell, and can stretch a film to bursting or (rarely) cause slow leakage by migrating through the joint between the film and the cell. Whenever there is a question about whether a SPEX SamplePrep X-Cell, window film, or combination

is that if the window film ruptures, the sample can spill and potentially contaminate or damage the spectrometer. However, an increasing number of samples are being run in so-called “normal optics” XRF spectrometers, where the X-ray source is above the cell instead of underneath. Bubbles trapped during assembly of the cell can collect against the window, displacing the sample and attenuating the X-rays reaching the sample and emitted by it. To use an X-Cell fitted with a SPEX SamplePrep Bubble-Free Cell Insert, assemble the cell, invert it, and tap it. Any bubbles should rise through the hole in the insert, and when the cell is turned right-side-up the bubbles will be trapped below the insert’s shelf, away from the window film. Most but not all bubbles get into an X-Cell during assembly; some samples outgas when heated by the X-ray beam. Tests have shown that a pinhole near the edge of the window film will allow such gases to escape without bulging or rupturing the film.The 3527I Bubble-Free Cell Insert coupled with the 3527 X-Cell will fit normal optics XRF spectrometers made by Diano, Rigaku/USA, Inc. and Shimadzu Scientific Instruments, Inc.

Selecting Thin Film Windows for XRF

An X-Cell liquid sample cell must be covered by a thin film which serves as the sample window. The film must effectively contain the liquid while allowing uniform transmission of the X-rays to the sample. Your choice of a window film depends on your analytical priorities and the nature of the sample. Is strength more important than X-ray transmission, or vice versa? Are there certain impurities you cannot tolerate in the film? Is the sample chemically aggressive, capable of weakening or penetrating certain films? Is it hot when you pour it into the X-Cell, or will it heat up during analysis and perhaps soften the film? Will it release volatiles and put pressure on the film from inside the cell?

Often the final choice of film balances several requirements. We recommend comparing films, and for that purpose we offer the SPEX-Pack, with samples of the films and X-Cells we supply. We also suggest strongly that if there is any question about the compatibility of a sample with certain films and/or X-Cells, you test them together outside the spectrometer. You are welcome to call us for further information at 1-800-LAB-SPEX x465.

When choosing a thin film window for your application, consider the following film characteristics:1. Strength: Leakage or rupture of the window during a run could cause damage to spectrometers with inverted optics. While SPEX SamplePrep accepts no responsibility for damage resulting from window leakage, the films offered by SPEX SamplePrep have been selected for their strength and durability. Kapton is the closest thing to a bombproof XRF window film. It is much stronger than any other SPEX SamplePrep film, heat-resistant to over 300° C, and chemically impervious to almost everything except strong alkali solutions. Six micron (6 µm) Mylar is also unusually strong for its thickness. The remaining films (5 µm Polypropylene, 4 µm Ultralene, and 3 µm Mylar ) have roughly comparable mechanical strengths, but differ in other key properties. Ultralene remains our best film in its balance of X-ray transmission, chemical resistivity, strength, and purity. All SPEX SamplePrep XRF window films except Kapton are weakened by heating much above 100° C.

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Chemical Resistance of Kapton® Percent Retained (typical)

Substance Conditions Tensile Strength Elongation Tensile Modulus

Acetic Acid, Glacial 36 days, 110°C 85 62 102

Acetone 365 days, R.T. 67 62 160

Benzene 365 days, R.T. 100 82 100

Chlorofrom 365 days, R.T. 93 74 96

p-Cresol 22 days, 200°C 100 77 102

Hexane 365 days, R.T. 97 89 100

Jet Fuel 190 days, R.T. 86 92 84

Methanol 365 days, R.T. 100 73 140

Sodium Hydroxide, 10% 5 days, R.T. - Degrades -

Toluene 365 days, R.T. 94 66 97

Transformer Oil 180 days, 150°C 100 100 100

Water: pH = 1 14 days, 100°C 65 30 100 pH = 4.2 14 days, 100°C 65 30 100 pH = 7.0 166 days, 100°C 65 20 100 pH = 8.9 14 days, 100°C 65 20 100 pH = 10.0 4 days, 100°C 60 10 100

Reprinted by permission of E.l. du Pont de Nemours & Co., Kapton,Summary of Properties E-50533.

thereof is likely to perform properly in a particular analytical situation, the analyst should first test these products in a way which does not risk contaminating the spectrometer. You are welcome to discuss any such problem with our applications specialists by calling (732) 623-0465.

Thin Film Transmission Characteristics

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Chemical Resistance of PolypropyleneSubstance Temperature (°C) Effect

Acetic Acid, Glacial 80 B

Acetic Acid, 50% 80 A

Acetone 60 A

Acetophenone 20 B

Ammonia, 30% 20 A

Ammonium Hydroxide, 10% 60 A

Amyl Acetate 20 B

Amyl Alcohol 60 B

Amyl Chloride 20 C

Aniline 60 A

Aqua Regia 20 B,E

Aviation Fuel 20 B

Benzene 20 B

Benzyl Alcohol 80 A

Brine 60 A

Bromine, Aqueous 20 C

Calcium Hypochlorite, 20% 60 B

Carbon Disulfide 20 B

Carbon Tetrachloride 20 C

Chlorine Gas 20 D

Chlorobenzene 20 C

Chloroform 20 C

Chromic Acid, 50% 60 A,E

Cyclohexanol 60 B

Cyclohexanone 20 B

Decaline 20 C

Detergents, 2% 100 A

Ethyl Acetate 60 B

Ethanol 80 A

Ethylene Glycol 60 A

Ethyl Ether 20 B

Chemical Resistance of Mylar® Percent Retained (typical)

Substance Conditions Tear Strength Elongation Tensile Strength

Acids Glcial Acetic Acid 24 hours, 75°C 100 100 100Hydrochloric Acid, 10% 100 100 75Hydroflouric Acid, Con. 0 0 0Nitric Acid, 10% 80 50 50Nitric Acid, 40% 75 50 75Sulfuric Acid, 30% 100 100 100Bases Ammonium 24 hours, 75°C 85 65 40Hydroxide, 2% 90 65 40Sodium Hydroxide, 10% 0 0 0Solvents Dioxane (1.4) 24 hours, 75°C 100 100 100Ethyl Acetate 100 100 100Ethanol 100 100 100Methyl Ethyl Ketone 100 100 100Toluene 100 100 100Trichloroethylene 100 100 100Miscellaneous Detergent, 25% 24 hours, 75°C 100 100 125Dimethyl Formamide 100 100 -Sodium Sulfide, 4% 72 45 40Tricresyl Phosphate 100 100 -Impregnants Hydrocarbon Oil, hi. vis. 900 hours, 100°C 92 88 87Hydrocarbon Oil, low. vis. (immersion) 96 67 123Silicone, DC 200 79 141 97Varnishes Asphalt Base GE9466 168 hours, 150°C 92 18 113Glyptal Resin GE2480 (baked) 90 80 60Phenolic Resin GE1678 92 3 73Silicone Resin GESR80 58 230 100Silicon Resin GESR28 100 54 97

Reprinted by permission of E.l. du Pont de Nemours & Co., Bulletin M-3C.

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Element Symbol

Actinium AcAluminum AlAmericium AmAntimony SbArgon ArArsenic AsAstatine AtBarium BaBerkelium BkBeryllium BeBismuth BiBoron BBromine BrCadmium CdCalcium CaCalifornium CfCarbon CCerium CeCesium CsChlorine ClChromium CrCobalt CoCopper CuCurium CmDysprosium DyEinsteinium EsErbium ErEuropium EuFermium FmFluorine FFrancium FrGadolinium GdGallium GaGermanium GeGold AuHafnium HfHelium HeHolmium HoHydrogen HIndium InIodine I

Element Symbol

Iridium IrIron FeKrypton KrLanthanum LaLawrencium LrLead PbLithium LiLutetium LuMagnesium MgManganese MnMendelevium MdMercury HgMolybdenum MoNeodymium NdNeon NeNeptunium NpNickel NiNiobium NbNitrogen NNobelium NoOsmium OsOxygen OPalladium PdPhosphorus PPlatinum PtPlutonium PuPolonium PoPotassium KPraseodymium PrPromethium PmProtactinium PaRadium RaRadon RnRhenium ReRhodium RhRubidium RbRuthenium RuSamarium SmScandium ScSelenium SeSilicon Si

Element Symbol

Silver AgSodium NaStrontium SrSulfur STantalum TaTechnetium TcTellurium TeTerbium TbThallium TlThorium ThThulium TmTin SnTitanium TiTungsten WUranium UVanadium VXenon XeYtterbium YbYttrium YZinc ZnZirconium Zr

Chemical Elements and Their SymbolsChemical Resistance of Polypropylene (con’t)Substance Temperature (°C) Effect

Formaldehyde 60 AGasoline 60 CHexane 60 BHydrochloric Acid, 30% 60 B,EHydrochloric Acid, 20% 60 AHydrogen Peroxide 60 BIodine Tincture 20 AKetones 20 ALinseed Oil 60 ALubricating Oil/Motor Oil 60 BNitric Acid, 70% 20 C,ENitric Acid, 10% 100 AParaffin 60 BPetroleum Ether 20 CPhenol 60 APlating Solutions 60 APyridine 20 ASilicone Oil 60 ASugars and Syrups 60 ASulfamic Acid 80 ASulfuric Acid, 98% 20 CSulfuric Acid, 50% 60 BSulfuric Acid, 10% 100 AToluene 20 CTrichloroethylene 20 CTurpentine 20 CWhisky 100 AWine 60 A

Code: A-Negligible Effect B-Limited Absorption or Attack C-Extensive Absorption or Permeation D-Extensive Attack E-May Produce Cracking Under StressReprinted by permission of Hercules Inc., Bulletin PPD-32E

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Section 3










2010 Geno/Grinder

8530 Shatterbox

3635 X-Press




Sec tion 3 :


Sec tion 3 :

Extraction of Nucleic Acids from Sugar Beet LeavesWith kind permission of the University of Kiel, Institute for Plant Cultivation and Plant Breeding

Within the framework of a TILLING project (Targeting Induced Local Lesions In Genomes) nucleic acids from leaf material are to be isolated from potential mutants. First a representative range of EMS (Ethyl methane sulfonate)-mutants is produced from 2000-5000 M2 plants and identified through TILLING. TILLING technology is a new and very effective method of reverse genetics for the production and identification of loss/gain of function-mutations of commercially valuable genes without genetic modification. To establish a TILLING platform with automated DNA extraction and PCR amplification of target genes, nucleic acid extraction of consistent yield and good quality is necessary. The effective detection of point mutation occurs via hetero duplex formation between DNA molecules of wild type and mutant.

ExperimentDuring the first project year, DNA from 2688 individual plants was isolated by using a 96- well micro titer plate kit. (NucleoSpin Multi- 96 Plant, Machery & Nagel, Düren). The plant tissue (0.5-1 g) was freeze dried in a 2ml Eppendorf vessel and then ground in the Geno/Grinder 2000 with two metal balls at a setting of 1000 (1000 strokes min-1) for 2 x 1 minute. For this, four mounts (plastic blocks) were produced in-house, each holding 24 Eppendorf vessels, and clamped into the Geno/Grinder. This type of sampling allows a flexible configuration of the individual samples. After grinding the tissue to a fine powder the samples were charged with 300 μl C1-buffer and homogenized in the Geno/Grinder for 30 seconds at a setting of 1000 (1000 strokes min-1).

Following lysis, the centrifuged samples are transferred into micro titer plates (round wells). After mixing, filtration, several rinse steps and drying the samples are finally eluted. DNA quantification is carried out by photometry and gel electrophoresis. Afterwards the samples can be stored long-term at -20°C.

YieldsApproximately 40% of the samples yielded between 100 and 1500ng DNA. Only 2% of the samples did not yield an adequate amount of DNA. The maximum yields were around 16 μg. For a routine PCR 1ng of highpure DNA is used. A further reduction of the DNA amount is possible. The selection of the mutant population is carried out with DNA pools. With a pool composition of 8 different samples the utilized DNA amount is reduced to 0.125ng/individual plant so that with 100ng DNA, the selection of DNA pools with 800 different target sequences becomes possible. The in vitro reproduction of linear DNA with GenomiPhi (GE Healthcare) allows further DNA analyses with small sample sizes.

Application Note SP011: DNA Extraction :: APPLICATION: Leaf Material

SP011

Mechanical Disruption for High-Throughput Fatty Acid Extraction from Animal Tissue SamplesWith kind permission of Sanofi-Aventis Germany GmbH

The mechanical homogenization for extraction of fatty acids is traditionally performed with an Ultra-Turrax in a buffer solution. However, both the purification and the slow sample preparation process in the analytical laboratory are labour-intensive. This method is no longer suitable for high sample throughput. Therefore it makes sense to combine the increased sample throughput with mechanical cell disruption. The use of 96-well micro titer plates and deep well plates is suitable for this process. The research in biotechnology/ gene technology also often demands the handling of a considerable number of samples. Sample preparation is often the bottleneck in the process of efficient analysis work. Up to now efforts to mechanically disrupt cells through grinding have been based on the modification of traditional ball or swing mills with an adapter for micro titer plates. The Geno/Grinder® 2000 is currently the first available mill specifically designed for cell disruption. The Geno/Grinder is a purpose-built ball mill that was originally conceived for the disruption of plant seeds but can also be used for animal and human samples.

Materials and MethodsTwo deep well plates (PP), used for collecting and storing of biological materials with 1.5 – 2 ml wells were placed next to each other in the Geno/Grinder clamp assembly. Freeze dried rat muscle or livers were placed in each plate well and spiked with 100mM KH2PO4 buffer at pH 2 or trichloro-acetic acid. Then two grinding balls were added. When all wells were filled, the plate was covered with a sealing mat. Thus prepared, the deep-well plates were clamped into the Geno/Grinder.

The samples were shaken vigorously for one minute at 1700 strokes per minute. This resulted in a fine, homogeneous sample dispersion within each well. Because of the vertical movement of the Geno/Grinder clamps, all wells are moved uniformly. This results in uniform and reproducible homogenization yields for all samples. As well as using titer plates with 96 wells it is also possible to use other formats, e.g. the “Biomek micro centrifuge 24 position tube rack” for Eppendorf Safe-Lock reaction tubes.

Results and DiscussionsThe results show that the Geno/Grinder 2000 is well suited for cell disruption of muscle and internal organs, i.e. liver. However, the quality of the results depends on the grinding accessories. Using several small Zirconium oxide beads with a diameter of 2mm does not give a satisfactory result. The sheer force is not sufficient to grind frozen muscle tissue. In contrast to this, it was possible to successfully homogenize the frozen material with stainless steel grinding balls of 4mm diameter.

Application Note SP001: Cell Disruption :: APPLICATION: Cell Lysis

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Perchloric Acid Extraction of Leaves for Measurement of Starch and Soluble MetabolitesWith kind permission of Dr. Marilyn Pike from The John Innes Centre, Department of Metabolic Biology, Norwich, UK

ExperimentThe harvested plant material (up to about 300 mg) is pre-weighed into 5ml polycarbonate vials (SPEX SamplePrep cat no. 2240- PC) suitable for use in 2000 Geno/Grinder® instrument. The tubes are then capped and placed immediately into liquid nitrogen (the time from harvest to freezing should be as short as possible) NB: Plant material can be harvested and frozen in any cryo-vial, then transferred to the Geno/Grinder tubes just before grinding if necessary.

Then transfer the tubes to dry ice to keep material frozen and add 500 μl ice cold 0.7M perchloric acid to each tube. Allow the acid to freeze. Place one good quality stainless steel ball, 10 mm diameter in 316 grade material, into each vial. Cap the tubes securely and place into the grinder foam holder.

Place and secure holder containing tubes into machine as per Geno/Grinder instructions.

Grind samples for 90 sec at 1500 strokes per minute (500 on the dial at x1 speed) — or until sample is smoothly pulverized. Transfer the tubes to ice immediately after grinding.

Add an additional 2500 μl ice cold 0.7M perchloric acid to each tube and vortex briefly. Centrifuge for 10 min at 4º C and14,000 xG.

Remove and neutralise the resulting supernatant to pH 6-7 by addition of 2 M KOH, 0.4 M MES, 0.4 M KCl. After centrifugation to remove the insoluble perchlorate, the neutralised solution can be used for assay of soluble metabolites. All steps should be carried out at 0-4 ºC.

The pellet contains the starch. After washing with water to remove the perchloric acid, it can be used for assay of starch according to Smith and Zeeman (2006).

ReferenceSmith AM and Zeeman SC (2006) Quantification of starch in plant tissues. Nature Protocols 1, 1342-1345.

Application Note SP013: Grinding / Disruption :: APPLICATION: Starch and Soluble Metabolite Measurement in Leaves

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Application Note SP011: DNA Extraction :: APPLICATION: Leaf Material

ConclusionHomogenization of sugar beet leaves for the purpose of extracting and isolating DNA for subsequent PCR amplification requires a sufficient amount of good quality nucleic acid yield (>1ng). With a basic grinding process of 2 x 1 minute grinds of dry leaf material and subsequent wet homogenization of 30 seconds only 2% of more than 2600 did not yield suitable DNA material. This demonstrates the excellent suitability of the Geno/Grinder 2000 for high throughput processing of sugar beet leaf material.

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Application Note SP014: RNA Extraction from Aspergillus parasiticus mycelium :: APPLICATION: RNA Extraction

Cryo-Station. After addition of Puresol the samples were homogenized in the SPEX SamplePrep 2000 Geno/Grinder for 2 minutes at 1050 strokes per minute. The sample tubes containing the homogenized sample buffer mix were transferred to a room temperature foam holder before homogenizing for an additional 2 minutes in the Geno/Grinder. During this final homogenization the Puresol buffer and ground mycelium thawed and mixed completely. Samples were removed from the Geno/Grinder and incubated at room temperature for 10 minutes before transferring to sterile 2 mL Eppendorf tubes. Total RNA was extracted according to manufacturer’s protocols, using the Aurum Total RNA fatty and fibrous tissue kit (Biorad).

YieldsTotal RNA was quantified using a ND-1000 Spectrophotometer (NanoDrop Technologies, Wilmington, DE) and visualized by gel electrophoresis (figure 1) to ensure quality. RNA concentration varied from 1.5 μg/uL to 3.8 μg/uL, in a final volume of 50 μL, with a A260/280 ratios varying from 2.01 to 2.19 or better. For a routine qRT-PCR 500 ng to 1 μg of total RNA is used as compared to 3 to 5 μg for the typical microarray experiment. More than adequate RNA for both microarray experiments and subsequent real time validation were isolated using this protocol.

ConclusionThough several environmental factors can affect the final quality of RNA, which directly affects both microarray and real time experiments, we demonstrate here a highly reliable methodology capable of isolating high quality RNA from small quantities of fungal mycelium. This protocol allows simultaneous processing of large numbers of samples, thus demonstrating the suitability of the Geno/Grinder for high throughput processing of fungal mycelium.

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Figure 1.1.0% Agarose gel. Lanes 1 and 14, 1 Kb ladder (Invitrogen). Lanes 2-5, replicates A-D of A. parasiticus SU-1 900 ng of total RNA isolated from Mycelium grown 48 hours in Yeast Extract Media. Lane 6-9, 10-13, 15-18, 19-22, and 23-26 replicates A-D of A. parasiticus SU-1 total RNA isolated from Mycelium collected at 3, 6, 12, 24 and 48 hours after shift from Yeast Extract Method to Yeast Extract Sucrose Media.

RNA Extraction from Aspergillus parasiticus myceliumWith kind permission of J.R. Wilkinson and R. Shivaji, Department of Biochemistry and Molecular Biology, Mississippi State University, P.O. Box 9650, Mississippi State, MS 39762, USA

IntroductionRecent genomic efforts on toxigenic and nontoxigenic Aspergillus species have advanced our understanding of the biology and genetics of these filamentous fungi. However, it is clear that these complex experiments suffer greatly from the variability in the quality of RNA between each replicate and it is critical to establish a platform to isolate high quality RNA for use in both microarray and qRT-PCR. In that context we describe here RNA isolation of A. parasiticus during a simple carbohydrate shift.

ExperimentFor this experiment A. parasiticus SRRC 143 (or SU-1; ATCC # 56775 and 201461) a wild-type strain used for biochemical and genetic studies in both B group (B1 and B2) and G group (G1 and G2) aflatoxins was chosen. Fresh spores were generated by plating 50 μL of 108 spores onto Difco Potato-Dextrose Agar (PDA) (American Scientific Products, Charlotte, NC) and incubated at 30ºC. The spores were collected from 5-day cultures with sterile 0.05% Triton X-100 (EMD Chemicals Inc.,

Gibbstown, NJ) A. parasiticus spores were inoculated to a final concentration of 105 spores/mL in 200 mL yeast extract (YE) liquid medium consisting of 25 g/L of yeast extract (DIFCO) and incubated for 48 hours at 30ºC with constant shaking at 150 rpm. The fungal mycelia were harvested by filtration in vacuo. The harvested mycelia were divided into 1 gram aliquots. One sample was frozen in liquid nitrogen and stored at - 80ºC for RNA preparation and the remaining aliquots were used to inoculate 100 mL of YES medium (60 g/L of sucrose and 25 g/L of yeast extract) and incubated at 30ºC with 150 rpm shaking until harvested at 3, 6, 12, 24, and 48 hours post-inoculation. The harvested fungal mycelium samples were flash frozen in liquid nitrogen and stored at -80ºC for RNA preparation and aflatoxin extraction.

Sample ExtractionBefore samples were processed in 24-well polyethylene vials (SPEX SamplePrep Cat. No. 2240-PE) containing one 3/8” (9.5 mm) stainless steel grinding ball per tube, they were arranged within a 24-well aluminum Cryo Adapter block (SPEX SamplePrep catalog 2263) and pre-chilled for 15 minutes in a Cryo-Station (SPEX SamplePrep Cat. No. 2600). Using a prechilled spatula 50 mg to 100 mg of the frozen fungal tissue was transferred to the pre-chilled tubes. The chilled samples and adapter block were clamped in the SPEX SamplePrep 2000 Geno/Grinder and ground at a rate of 1050 strokes per min (dial setting of 150) for 1 minute and 30 seconds. The samples and block combination were quickly removed from the Geno/Grinder and returned to the Cryo-Station. To each ground sample 1 mL of Puresol from the Aurum Total RNA fatty and fibrous tissue kit (Biorad) was added. To ensure that all ground tissue was suspended in Puresol the samples still housed within the adapter block was inverted 2-3 times and returned to the

Application Note SP014: RNA Extraction from Aspergillus parasiticus mycelium :: APPLICATION: RNA Extraction

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Effect of Lysis Time and Other Variables on DNA Extraction from Fresh Basil Lysed in 2 ml Tubes With the Geno/Grinder®With kind permission of Sunrise Science, San Diego, CA

ABSTRACTMechanical cell lysers are used for effective extraction of genomic DNA from samples containing plant or animal tissue. However, all mechanical lysers do not provide the same outcome. The 2000 Geno/Grinder® was compared with a competitive cell lyser (Competitor A) for extraction of DNA from fresh basil. In addition, homogenization time and operating rate were varied for the Geno?Grinder, while buffers, grinding media and tube size remained constant. Results indicated that use of the Geno/Grinder provided DNA with higher molecular weight than the competitive instrument. In addition, optimal homogenization conditions using the Geno/Grinder were found to be 90 sec. at a rate of 2000 cyc/min.

SAMPLE SETUP: : TUBES: 2 ml conical-bottom screw-cap tubes on a standard 48 well rack.

: : GRINDING MEDIA: Garnet and single 1/4 inch ceramic bead (“Lysing Matrix A”).

: : SAMPLE: 100 mg fresh basil leaf, harvested from live plant immediately prior to lysis.

>1 gram of leaves were chopped into small pieces and mixed. 100 mg of chopped leaf mixture was placed into each of 10 tubes.

: : LYSIS REAGENTS: 400 ml of Buffer Ap1 and 4 ml of RNaseA from commercially available genomic DNA isolation kit were added to each tube.

: : HOMOGENIZATION:

Tube # 1 2 3 4 5 6 7 8 9 10

Homogenizer* CA CA GG GG GG GG GG GG GG GG

Time (sec) 40 40 40 40 60 60 90 90 120 120

Speed (cyc/min) 6000 6000 2000 2000 2000 2000 2000 2000 2000 2000

*CA = Competitor A Instrument *GG = Geno/Grinder® Instrument (SPEX SamplePrep LLC)

Application Note SP017: Lysis Time and Other Variables on DNA Extraction :: APPLICATION: DNA Extraction from Fresh Basil Lysed

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Quick DNA Extraction from Rice SeedWith kind permission of RiceTec Inc., Alvin, TX

IntroductionDNA extractions can be a very time consuming and tedious process. Finding a quick method in which DNA could be extracted and used for PCR is essential. Described below is a quick “dirty” method that produces a high enough concentration of DNA that can be used for PCR.

Sample ExtractionSamples are prepared using a 96-well 1 mL assay block. Dispense one 5/32 in. (4 mm) stainless steel bead into each well using the Grinding Ball Dispenser (SPEX SamplePrep Cat. No. 2100). Next, add one seed to each well. Dispense extraction buffer into each well and securely cap each well. After the samples have been capped, grind them in the SPEX SamplePrep 2000 Geno/Grinder at 500 strokes/minute for two minutes. Centrifuge for 1 minute to bring all liquid to the bottom of the assay block. Incubate the samples in about 1in. (25.4 mm) of water at 95ºC for 20 minutes then place them on ice for approximately 10 minutes or until samples are cool to the touch. Centrifuge again for 1 minute. Add neutralizing extraction buffer and seal the assay block with sealing film. Centrifuge the samples for 10 minutes at 3000 rpm. Transfer 300 μL of the supernatant to a clean 96-well plate. DNA can be further purified with clean-up kits available on the market.

YieldsThe total concentrations yielded from the samples range from 3-7 ng/μL in a final volume of 200 μL with purities of 1.5-1.8.

ConclusionThe method described above is sufficient for PCR and it takes less time than the standard chloroform extraction. Total time ranges from one to two hours.

Application Note SP016: Lysing / Homogenization :: APPLICATION: DNA Extraction from Rice Seeds

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High Throughput Disruption of Yeast in a 96-Well Format

The wealth of information generated from years of biochemical, genetic, and molecular analyses has made yeasts both model biological systems and tools for biopharmaceutical scientists. Consequently, yeast are a popular host for gene expression studies and for the production of recombinant proteins. Though many yeast species are in use, including Pichia, Hansenula, and Debaryomyces, the most popular yeast continues to be Saccharomyces.

Yeast mRNA and intracellular proteins are often times difficult to extract intact from cells by traditional enzymatic methods. Lysing enzymes are often crude preparations containing RNases and proteases that will not only attack the cell wall, but also the molecules of interest. Furthermore, protoplasts generated from enzymatic digestion usually require lysis with detergents that will also denature many proteins to inactivity. Therefore, mechanical cracking/fracturing of the yeast cell is often required to liberate the molecules.

Mechanical disruption of yeasts has traditionally been accomplished by using either a press or bead mill (i.e., bead beater). In both approaches, samples are processed individually. For experiments where large numbers of yeast clones must be examined in a high throughput screening environment, individual processing is a major bottleneck and impractical. Consequently, a method is needed that combines mechanical disruption of cells in a high throughput format. The Geno/GrinderTM (SPEX SamplePrep, Metuchen, NJ), a bead mill originally designed to smash seeds in deep well plates, can be used to disrupt yeast in a microwell plate format.

Materials and MethodsSaccharomyces cerevisiae was cultured overnight in YPD medium (1% yeast extract, 2% peptone, 2% glucose) at 30°C with agitation (150 rpm). Using a sterile 96 well microtiter plate (Corning Life Science Products), 100 ml of culture broth was added to each well along with 50 mg of acid washed glass beads (Sigma, G-8772). A range of glass bead sizes was used including 106 mesh (very small), 150-212 mesh, 212-300 mesh, and 425-600 mesh (large). Culture broth without glass beads was also prepared as a

Application Note SP018: High Throughput Disruption of Yeast in a 96-Well Format :: APPLICATION: DNA/RNA and Other Extractions

Figure 1. Negative Control yeast without glass beads. The yeast remained intact after processing for 10 min. in the Geno/Grinder.

SP018

DNA PURIFICATIONPost-lysis purification was performed using a commercially available plant kit according to the kit manual.

DATA

Outside Marker (lanes 1 and 14): Kb ladder Inside Marker (lanes 2 and 13): Lambda Hind III

INTERPRETATIONAfter homogenization for 40 sec. using the competitive cell lyser, DNA of lower molecular weight (lower quality) was obtained than for samples processed at any of the four run times using the Geno/Grinder.

At both 90 and 120 sec., nearly all the DNA isolated using the Geno/Grinder was higher than 6,557 bp, indicating that significant shearing or degradation had not occurred, even at the longer processing time. Little to no difference in the results was observed at these two processing times. While run times for 40 and 60 sec. on the Geno/Grinder yielded high molecular weight DNA, insufficient quantities were obtained.

CONCLUSIONSThe best conditions for processing fresh basil were found to be 90 sec. at 2000 cyc/min with the 2000 Geno/Grinder. Increasing the processing time to 120 sec. did not appear to offer any benefit. In addition, use of the Geno/Grinder gave higher quality DNA than was obtained using the competitive lyser.

Further optimization of the processing time may be possible by varying the type and size of grinding media. However, that was beyond the scope of this project and was not attempted.

Optimal lysing conditions for the re-designed 2010 Geno/Grinder may differ somewhat from those reported here.

Application Note SP017: Lysis Time and Other Variables on DNA Extraction :: APPLICATION: DNA Extraction from Fresh Basil Lysed

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Comparison of Methods for the Isolation of DNA from SoybeansWith kind permission David Burden, Ph.D., President of BT&C Incorporated

The isolation of nucleic acids from intact seeds requires mechanically disrupting the seed followed by the extraction and subsequent purification of the nucleic acid. The mechanical disruption is often performed manually with a mortar and pestle, an approach that is not practical for high throughput screening of seeds as manual grinding is slow and reuse of mortar and pestles may lead to cross-contamination. Alternatively, nucleic acids can be isolated from seeds in a microwell plate format using a ball mill that mechanically disrupts the seeds. Conventional isolation methodologies can then be used to purify the nucleic acids from the seed homogenates.

The efficiency of seed disruption is dependant upon the type of ball mill used in the grinding process. Standard bead mills adapted to microwell plates are modeled after “paint shakers” and move the plates in a “figure-eight” motion. This motion does not lead to uniform seed disruption. The Geno/Grinder® is designed to effectively disrupt cellular materials by oscillating the plate vertically. This motion allows balls to impact the seeds more directly than standard mills where balls impact the well walls in addition to the seed. Soybeans that are soaked overnight in water are effectively and uniformly homogenized in less than 3 minutes. The resulting pulp can then be used as a source of DNA for genetic analysis.

Materials and MethodsSoybeans were placed in each 2.0 mL deep-well of a 96-well plate containing a 4 mm stainless steel grinding ball (Cat. No. 2150) and soaked for 12 hours in distilled water prior to processing. Using a grinding ball dispenser a grinding ball was also placed on top of each seed. The plate was sealed with a fitted Teflon®/silicone mat and placed in the Geno/Grinder. A piece of adsorbent paper was placed on top of the plate and the plate was locked into the grinder. The seeds were disrupted for 2.5 minutes at 1500 rpm. The deep well plate with lid was then centrifuged at 1500 rpm to pellet lysate and condense liquid from the rim and walls of the well. Without centrifugation, the probability of well to well cross contamination of genetic material is greatly increased. Once centrifuged, the lid is carefully removed.

Four different DNA methods were compared for the isolation of DNA from the soybean homogenates, namely the CTAB, Wizard Genomic, Qiagen Plant DNeasy, and Qiagen Genomic methods. Each of these methods is summarized briefly below.

Application Note SP019: Comparison of Methods for the Isolation of DNA from Soybeans :: APPLICATION: DNA/RNA and Other Extractions

SP019

negative control. The plate was sealed with a rubber-sealing mat and locked into the Geno/Grinder. The yeast cells were disrupted for 5 and 10 minutes at 1500 rpm. Following disruption, the yeast cultures were examined by phase contrast microscopy and photographed.

Results and DiscussionThe efficiency of yeast disruption is dependent upon the size of the glass beads and the duration of the grinding. Figures 1-3 demonstrate the efficiency of cell disruption with 425-600 mesh glass beads at 5 and 10 minutes. Lower mesh glass beads were less effective in cracking cells with the 106 mesh beads being particularly ineffective. It is concluded that 425-600 mesh glass beads are most suited for disrupting Saccharomyces.

The design of the Geno/Grinder is also believed to be a factor in the disruption process. Standard bead mills adapted to microwell plates are modeled after “paint shakers” and move the plates in a “figure eight” motion. This motion is not believed to lead to uniform cell disruption. The Geno/Grinder is designed to effectively disrupt cellular materials by oscillating the plate vertically. This motion allows glass beads to impact the cells more directly than standard mills where beads impact the well walls in addition to the cells.

Application Note SP018: High Throughput Disruption of Yeast in a 96-Well Format :: APPLICATION: DNA/RNA and Other Extractions

Figure 2. Yeast disrupted for 5 min. with 425-600 mesh glass beads. Cracked yeast appear as dark “ghosts” while intact yeast continue to refract light and appear as bright.

Figure 3. Yeast disrupted for 10 min. with 425-600 mesh glass beads. Most yeast have been effectively disrupted as is demonstrated by cell ghosts and cellular debris. Some intact yeast remain.

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DNA Purification Using a Qiagen Genomic ColumnAfter grinding, the soy was suspended in 10 mL of Buffer G2. A Qiagen genomic tip 100 column was equilibrated with Buffer QBT. Following equilibration, the soy sample was applied to the column, and the column was washed two times with Buffer QC. Elution of the DNA was done with 5 mL Buffer QF. The DNA was precipitated with 0.7 volumes of isopropanol and freezing at -80° C for 20 minutes. The sample was thawed at room temperature and centrifuged to pellet the DNA. The pellet was washed with 70% ice-cold ethanol and then centrifuged. The ethanol was removed, and any residual ethanol was removed by centrifuging in the DNA Speed Vac. The DNA was then resuspended in 50 µL Buffer TE.

A comparison of the DNA yields for the isolation procedures was made by agarose gel electrophoresis (0.7% agarose in 1X TAE buffer) which is illustrated in Figure 1.

The genomic DNA was tested for genetically modified sequences, i.e., GMO analysis, using PCR. Samples were analyzed for the soy lectin gene and Cauliflower Mosaic Virus 35S promoter/Petunia transcription sequence marker found in genetically modified soybean (Roundup ReadySoya). Analysis of the reactions was done electrophoretically (1X TAE with 3% agarose) and is illustrated in Figure 2.

Results and DiscussionThe isolation method used to extract DNA from soybean homogenate affects the quantity and quality of the genomic DNA isolated. Agarose gel electrophoresis of the genomic DNA (Fig. 2) illustrates a significant difference in yield and fragment size of the DNA. Based on the fluorescence of the DNA smear in the ethidium bromide stained gel, the CTAB method produced the greatest yield, but fragment size is extremely small as compared to the other DNA preparations and measured against the molecular weight markers. Depending upon the application of the DNA, lower yields of large sized fragments (e.g., the Wizard method or Genomic Column method) may be preferred.

PCR analysis of the samples was positive for the soy lectin gene and negative for the 35 S promoter/Petunia TS sequence. This analysis demonstrates that soybean homogenates generated from the Geno/Grinder are suitable for genetic analysis. This is particularly useful for high throughput PCR analysis in agricultural biotechnology laboratories or laboratories performing GMO analysis.


Figure 1. Comparison of methods for DNA isolation from Soybean. Lanes 1, 4 and 7 – Lambda HindIII digest, Lane 2 – Qiagen Genomic DNA Kit, Lane 3 – CTAB Methood, Lane 5 – Promgea Wizard Kit, Lane 6 – Qiagen DNeasy Kit.

Figure 2. PCR analysis of isolated DNA from soybean

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DNA Purification Using the Qiagen DNeasy™ Plant KitAfter grinding, the soy was mixed with 400 µL Buffer AP1 and 4 µL RNase stock solution. The mixture was incubated for 10 minutes at 65° C. Buffer AP2 (130 µL) was then added to the lysate, mixed, and incubated for 5 minutes on ice. The precipitates were removed with the QIAshredder™ spin column. The eluent was mixed with 0.5 volume of Buffer AP3 and 1 volume of ethanol (96-100%). After mixing, the lysate was applied to the DNeasy mini spin column and centrifuged at 9000 rpm for 1 minute. During this step, the DNA binds to the column. The flow-through is discarded. The column was washed with Buffer AW, and then centrifuged at maximum speed in order to dry the membrane. The DNA was eluted with 50 µL Buffer AE by incubating for 5 minutes at room temperature and then centrifuging for 1 minute at 9000 rpm.

DNA Purification with the Promega Wizard® Genomic DNA Purification KitAfter grinding, the soy was mixed with 600 µL Nuclei Lysis Solution and incubated for 15 minutes at 65° C. RNase was added followed by a15 minute incubation at 37° C and a 5 minute cooling to room temperature. Protein Precipitation Solution was added followed by centrifugation to pellet precipitated proteins. The supernatant containing the DNA was transferred to another microfuge tube containing isopropanol. The sample was mixed gently by inversion and then centrifuged at 9000 rpm. The supernatant was removed and the pellet was washed with 70% ethanol. The tube was centrifuged and the ethanol was decanted. Excess ethanol was removed by inverting the tube onto absorbent paper and air-drying for 15 minutes. The DNA was resuspended in 50 µL DNA Rehydration Solution during incubation for 1 hour at 65° C.

CTAB Method for DNA PurificationThe ground soy was diluted in 500 µL CTAB buffer (20 g CTAB/L, 1.4 M NaCl, 0.1 M Tris/HCl, 20 mM EDTA). This mixture was incubated for 30 minutes at 65° C. It was then centrifuged for 10 minutes at 9000 rpm. The upper layer was extracted with an equal volume of chloroform. After mixing for 30 seconds, the mixture was centrifuged for 10 minutes at 9000 rpm. The supernatant was transferred to a new tube, and two volumes of CTAB precipitation solution (5 g CTAB/L, 0.04 M NaCl) were added. The mixture was incubated for 60 minutes at room temperature and then centrifuged for 5 minutes at 9000 rpm. The supernatant was removed, and the precipitate was dissolved in 1.2 M NaCl. This solution was chloroform extracted. The upper layer (aqueous phase) was transferred to a new tube and 0.7 volumes of isopropanol was added. The solution was then frozen for 20 minutes at -80° C. The sample was thawed at room temperature and centrifuged. The resulting DNA pellet was washed with ice-cold 70% ethanol and centrifuged. The ethanol was discarded, and residual ethanol was removed with a DNA SpeedVac. The DNA pellet was resuspended in TE buffer.


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Experiments were performed with 250 – 500 ml of bacteria: in one case the gram-negative, salt-tolerant Halomonas elongata, and in the other, gram-positive Bacillus. Both organisms were grown to late log state at 370 C, with shaking, then harvested via centrifugation at 7000 RPM for 10 minutes. Cells were washed once with sterile 8% (w/v) NaCl solution to remove spent growth medium. After washing, the cell pellet was suspended in 20 ml of sterile 8% NaCl solution. Each well of the test plate was filled with 0.5 ml of this suspension; it was important that the cells to be lysed were present in high concentration. 0.4 grams of silica beads were added to each well, and the plate was sealed. The filled titer plates were then shaken in the Geno/Grinder at a setting of 1450 strokes per minute, in one-minute intervals for periods ranging from one minute to twelve minutes. Culture filtrate from the shaken plates was then measured for optical density at 260 nm. The average data for three separate “well sets” is shown in Figure 1.

ConclusionThe 2000 Geno-Grinder is capable of lysing bacterial cells in 96-well titer plates with 400-600 μ silica beads as grinding media. With increased grinding time culture filtrate shows a definite increase in optical density, indicating release of nucleic acids during grinding. Grinding times of 6 to 9 minutes appear to be suitable for producing measurable amounts of nucleic acid.

Application Note SP020: Lysing of Bacterial Cells in the Geno/Grinder :: APPLICATION: DNA/RNA and Other Extractions

SP020

Lysing of Bacterial Cells in the Geno/GrinderWith kind permission of Russell H. Vreeland, West Chester University, January 2003

The SPEX SamplePrep 2000 Geno/Grinder was tested to determine if this technology could be used to lyse bacterial cells. Standard 96-well titer plates were used with 400-600 μ silica grinding beads (Molecular Biology Grade, cat. no. 2166).

The delivery of the beads into each cell of the titer plate can be accomplished in a number of ways. In this case micropipette tips were filled to the mark with grinding beads, and each tip emptied into a titer plate well. This technique will deliver approximately 0.4 grams of silica beads per well. Other delivery systems are commercially available.

Application Note SP020: Lysing of Bacterial Cells in the Geno/Grinder :: APPLICATION: DNA/RNA and Other Extractions

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Each real-time TaqMan PCR reaction contained 400 nM of each primer, 80 nM of the TaqMan probe and commercially available PCR mastermix (TaqMan Universal PCR Mastermix, Applied Biosystems) containing 10 mM Tris-HCl (pH 8.3), 50 mM KCl, 5 mM MgCl2, 2.5 mM deoxynucleotide triphosphates, 0.625 U AmpliTaq Gold DNA polymerase per reaction, 0.25 U AmpErase UNG per reaction and 5 µl of the diluted cDNA sample or the gDNA in a final volume of 25 µl. The samples were placed in 96 well plates and amplified in an automated fluorometer (ABI PRISM 7700 Sequence Detection System, Applied Biosystems). Amplification conditions were 2 min at 50°C, 10 min at 95°C, 40 cycles of 15 s at 95°C and 60 s at 60°C.

ReferencesGAPDH pseudogenes:

Galland, F., M. Stefanova, V. Pirisi, and D. Birnbaum. 1990. Characterization of a murine glyceraldehyde-3-phosphate dehydrogenase pseudogene. Biochimie. 72:759-62.

Garcia-Meunier, P., M. Etienne-Julan, P. Fort, M. Piechaczyk, and F. Bonhomme. 1993. Concerted evolution in the GAPDH family of retrotransposed pseudogenes. Mamm. Genome. 4:695-703. Leutenegger, C.M. , B. von Rechenberg, K. Zlinsky, C. Mislin, M. Akens, J. Auer, and H. Lutz:

Quantitative real-time PCR for equine cytokine mRNA in nondecalcified bone tissue embedded in methyl methacrylate. Calcified Tissue International, 65:378-383, 1999.

Leutenegger, C.M., A.M. Alluwaimi, W. Smith, L. Perani, J.M. Cullor. Quantitation of bovine cytokine mRNA in milk cells of healthy cattle by real-time TaqMan polymerase chain reaction. Veterinary Immunology and Immunopathology. 77: 275-287, 2001.

Leutenegger, C.M.. The real-time TaqMan PCR and applications in Veterinary Medicine. Veterinary Sciences Tomorrow, Online Journal, Jan 1, 2001.

Application Note SP021: Extraction of RNA/cDNA and Genomic DNA from Tissue with Real-Time PCRAPPLICATION: DNA/RNA and Other Extractions

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Extraction of RNA/cDNA and Genomic DNA from Tissue with Real-Time PCRWith kind permission of Christian M. Leutenegger, Ph.D., University of California, Davis, [email protected], January 2003

Sample Collection, Preparation and Tissue GrindingFresh samples of animal tissue were collected, trimmed to approximately 50 – 100 mg of wet weight, snap-frozen in liquid nitrogen, and stored at –800º C. The animals were man, dog, cat, mouse, cow, and horse, as well as fish and clams; see Table 1. Before DNA and/or RNA extraction, the tissues were transferred frozen to a deep-well titer plate standing on a block of dry ice. Each well contained two 4-mm stainless steel balls (SPEX CertiPrep cat. no. 2150) and 500 microliters of buffer (Applied Biosystems nucleic acid purification lysis buffer). The plates were sealed with a plastic cover and subjected to grinding in the 2000 Geno/Grinder for 2 minutes at a setting of 1000 strokes per minute. After 30 minutes at 40º C, lysates were used for either gDNA extraction or total RNA extraction. Conditions were optimized for an Applied Biosystems 6700 automated nucleic acid (ANA) workstation, according to the manufacturer’s instructions. The final amount of tissue subjected to RNA and/or DNA extractions was between 10 and 20 mg.

Quality Control of Extracted RNA/cDNA and Genomic DNA Using Real-Time Taqman® PCRTo assess the quality of the extracted RNA, complementary DNA (cDNA) was synthesized using Invitrogen products: 200 units of SuperScript III, 600 ng of random hexadeoxyribonucleotide (pd(N)6) primer (random hexameter priner), 10 U RnaseOut (Rnase inhibitor), and 1 mM dNTPs in a final volume of 40 l. The reverse transcription reaction took place for 50 minutes at 500 C. After addition of 60 µl of water, the reaction was terminated by heating for 5 minutes to 95ºC and cooling on ice. The quality of the cDNA is judged according to the CT value obtained with a endogenous control TaqMan PCR system from a defined amount of tissue. Samples with values above a certain threshold indicate impaired sample quality and degradation of total RNA and warrant re-extraction of a back-up sample. To assess the quality of cDNA, we normally use TaqMan PCR systems targeting species specific glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or ribosomal genes (such as 18S rRNA or ITS-2).

To assess the quality of extracted genomic DNA, species specific TaqMan PCR systems were developed targeting single copy genes to allow the quantitation of genome equivalents and cell numbers. An overview of TaqMan systems used for gDNA quality control is given in Table 1. The gDNA quality from a defined amount of tissue is judged according to the CT value using a TaqMan PCR system targeting a single copy gene. Samples with values above a certain threshold indicate degraded DNA and warrant re-extraction of a back-up sample. GAPDH TaqMan systems can be used to target the single copy GAPDH pseudogene (Galland et al., 1990; Garcia-Meunier et al., 1993).


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Table 2: Composition of real-time fluorogenic Taqman PCR assay

GAPDH IL4 IL10 IL 12 p35 IL 12 p40 IFN-g Il16

2x Mastermix 12.5 12.5 12.5 12.5 12.5 12.5 12.5

10x Buffer A 1x 1x 1x 1x 1x 1x 1x

MgCl2 (mM) 5 5 5 5 5 5 5

dATP (mM) 300 300 300 300 300 300 300

dCTP (mM) 300 300 300 300 300 300 300

dGTP (mM) 300 300 300 300 300 300 300

dUTP (mM) 300 300 300 300 300 300 300

Forward primer (nM) 400 400 400 400 400 400 400

Reverse primer (nM) 400 400 400 400 400 400 400

TaqMan probe (nM) 80 80 80 80 80 80 80

AmpliTaq Gold (U) 0.75 0.75 0.75 0.75 0.75 0.75 0.75

AmpErase UNG (U) 0.25 0.25 0.25 0.25 0.25 0.25 0.25

Template (cDNA) 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml

H2O Ad 25 ml Ad 25 ml Ad 25 ml Ad 25 ml Ad 25 ml Ad 25 ml Ad 25 ml

10 x TaqMan Buffer A contains 500 mM KCl, 100 mM Tris-HCl, 100 mM EDTA, 600 nM, passive reference ROX, pH 8.3 at room temperature


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Table 1: Overview of tissue samples, species and TaqMan systems used to assess quality of cDNA and gDNA

Tissue Species RNA/cDNA Genomic DNA Target Gene CT Value Target Gene CT Value

Skin Dog GAPDH <21 GAPDH <24

Skeletal Muscle Dog GAPDH <24 GAPDH <25

Lung Cow GAPDH <24 GAPDH <25

Brain Cat GAPDH <24 CCR5 <25

Lymph Node Cat GAPDH <21 CCR5 <22

Spleen Cat GAPDH <21 CCR5 <22

Thymus Cat GAPDH <21 CCR5 <22

Tonsil Cat GAPDH <21 CCR5 <22

Cartilage Horse GAPDH <24 IGF-I <26

Heart Mouse GAPDH <24 IGF-I <24

Tail Mouse GAPDH <21 IGF-I <21

Fin Rainbow Trout ITS-2 <14 IGF-I <26

Gill Rainbow Trout ITS-2 <14 IGF-I <24

Spine Tissue Rainbow Trout ITS-2 <14 IGF-I <24

Mussel Rainbow Trout ITS-2 <14 IGF-I <26

Liver Rainbow Trout ITS-2 <14 IGF-I <21

Cranium Rainbow Trout ITS-2 <16 IGF-I <24

Gill Koi ITS-2 <14 Glucokinase <24

Liver Koi ITS-2 <14 Glucokinase <21

Digestive Gland Clams 18S rRNA <11 NA NA

Mussel Clams 18S rRNA <11 NA NA

Liver Human GAPDH <21 IL-2 <21

Skeletal Muscle Human GAPDH <24 IL-2 <24

Kidney Human GAPDH <24 IL-2 <24

NA: Not Available

CT Value: Cycle Threshold Value: PCR cycle at which the fluorescent intensity exceeds the threshold


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Sample PreparationSample preparation for pesticide residue analysis has typically followed DuPont Report No. AMR 3705-95, “Analytical Method for the Determination of Famoxadone and Cymoxanil Residues in Various Matrices.” (2) In this procedure, ground-up samples are weighed into extraction jars, followed by the addition of water to allow them to re-hydrate prior to extraction. Acetonitrile is added and samples are ground with a laboratory homogenizer. The plant matrix is then allowed to settle-out and liquid extracts are filtered and collected in mixing cylinders containing sodium chloride. The mixing cylinders are capped, shaken, and inverted to aid in the dissolution of sodium chloride and then allowed to stand while the acetonitrile (upper layer) and water phases separated. Separate acetonitrile aliquots are taken for cymoxanil and famoxadone analysis.

Cymoxanil and Famoxadone AnalysisFor the analysis of cymoxanil, the acetonitrile aliquot is back extracted with hexane, concentrated, diluted with water and passed through a conditioned strong anion exchange (SAX) solid phase extraction (SPE) cartridge stacked on top of a conditioned carbon black SPE cartridge. Cymoxanil passes through the SAX cartridge and is retained on the carbon black cartridge. It is then selectively eluted off the cartridge and the concentrated cymoxanil residues are dissolved in a hexane/ethyl acetate mixture. The resulting solutions are passed through silica SPE cartridges. Cymoxanil is retained, selectively eluted, and concentrated into a methanol/water mixture adjusted to pH 3. Samples are filtered and analyzed by HPLC with UV detection.

For the analysis of famoxadone, the acetonitrile aliquot is back extracted with hexane, the acetonitrile fraction is concentrated to approximately 2 mL and carefully taken to dryness. The residue is dissolved in 10% ethyl ether/90% hexane (v/v) and passed through a glass chromatography column packed with a layer of sodium sulfate, Florisil, and sodium sulfate. Each column is washed with additional 10% ethyl acetate/90% hexane (v/v). The eluate is concentrated, carefully taken to dryness, reconstituted in acetonitrile and water and analyzed by LC/UV.

Sample Prep for LC/MS AnalysisThere are approximately 20 clean-up steps in the AMR 3705-95 procedure, which limits the number of samples to 8-10 per day. A modification to this procedure, laid out in DuPont 13753 -“Analytical Method for the Determination of Cymoxanil and its Metabolites in Leafy Vegetables Using LC/MS” (3), significantly reduces the number of clean-up steps. In this procedure, samples are extracted using an acetonitrile/water mixture as described in DuPont Report No. AMR 3705-9, Revision 2. For cymoxanil, NaCl is added to an aliquot to separate the aqueous phase from the organic phase. The aqueous phase is discarded, and the acetonitrile layer containing cymoxanil is passed through a SAX SPE column. The extract is then cleaned further using a hexane liquid/liquid extraction followed by an Envi Carb SPE column. Cymoxanil is not retained on either of these columns, so the eluent is passed directly into an LC/MS system for analysis. Based on the sensitivity and selectivity of LC/MS, the number of sample clean-up steps is reduced to approximately 10, allowing for the analysis of up to 16 samples per day.

APPLICATION NOTE SP022: The Benefits of the Geno/Grinder for Pesticide Residue AnalysisAPPLICATION: Pesticide Residue Extraction

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The Benefits of the Geno/Grinder® High-Throughput Tissue Homogenizer to Increase Sample Throughput for Pesticide Residue Analysis by LC/MS/MSWith kind permission of: Joseph McClory, E. I. DuPont de Numours and Co., Inc., Keith Tucker, SPEX SamplePrep LLC, Robert Thomas, Scientific Solutions

SummaryThe high-throughput analysis of pesticides has been hindered by the slow and laborious sample preparation stage. Using traditional clean-up procedures, over twenty steps are required to get the sample in a convenient form for analysis, which limits sample throughput to typically 8 samples per day. This study evaluates a new approach using an innovative laboratory tissue homogenizer, to extract various pesticide residues from different kinds of plant materials and to identify and quantitate the active ingredients by LC/MS/MS. It shows that by automating the sample preparation procedure, the number of clean-up steps can be significantly reduced, resulting in an increased sample throughput over traditional approaches.

DiscussionThree of the leading pesticides sold by DuPont are Famoxate®, Curzate® and Tanos® to control various fungal diseases in crops. Famoxate is a fungicide containing the active ingredient famoxadone and is mainly used to control early blight tomatoes. It protects from spore germination and fungal growth by its strong adhesion to leaf and stem surfaces. Curzate is a fungicide that contains cymoxanil as the active ingredient and is mainly used for late blight potatoes, but is also applied to grapes, tomatoes, cucumbers and leafy vegetables. Its protection mechanism is to penetrate the surface to induce host defense response to stop lesion growth and sporulation. Tanos is a fungicide containing both famoxadone and cymoxanil and the cumulative effect of these two active ingredients offers even better control of late blight potatoes. The typical pesticide residue set at DuPont contains 8 samples and usually takes an 8 hour workday to complete. The goal of this study was to take advantage of recent developments in sample preparation with the 2000 Geno/Grinder® and instrument sensitivity and selectivity using the API 5000 LC/MS/MS system to increase the number of samples analyzed per day by a factor of three. The methodology follows OPPTS 860-1360 Multiresidue Test Method (1), which has been developed by the United States Environmental Protection Agency (USEPA) Office of Prevention, Pesticides and Toxic Substances (OPPTS), for use in the testing of pesticides and toxic substances. Under this method, test data must be submitted to the Agency for review under Federal regulations and dictates that average recoveries have to be 70-120% with a coefficient of variation (CV) or RSD of less than 15%.


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quadrupole, which is slightly pressurized by the introduction of a collision gas like hydrogen or helium. In the MS/MS mode, the first quadrupole is used in a mass- resolving mode to select the precursor ion. The second quadrupole (or pressurized multipole collision cell) is used to produce fragmentation of the precursor or parent ion. The final quadrupole is used in a mass-resolving mode to provide mass analysis of the resulting fragmented or daughter ions. These species are then compared to reference spectra/data in order to produce unambiguous identification of the biomolecules of interest. The LC/MS/MS instrument used for this study is shown in Figure 2.

The benefit of the MS/MS design is that it can also be used in MS mode for quantitation purposes. In this mode, the first quadrupole is typically used in the rf-only mode, as a wide bandpass filter to transmit molecular ions of a wide mass range. The collision cell is also used in the rf-only mode, but this time no collision gas is flowing, so the cell is just used to transmit ions to the last quadrupole, which is used in the mass resolving mode. The ability to switch between both modes is very important, in order to maximize the amount of data being generated, particularly when small amounts of sample are being analyzed. There is no question that the higher number of daughter or fragmented ions that are being identified, will lead to better confirmation of the parent molecules. This is especially important when analyzing data from an LC separation. Cymoxanil, for example, which is an aliphatic nitrogen compound with a formula of C7H10N4O3 and molecular weight of 198

typically elutes off the column after 10-15 minutes, over a period of 20-30 seconds. For this reason it is very important to be able to switch very rapidly between SIM and MRM mode, to collect both the molecular and fragmented information, for positive and unambiguous identification of these species. Once identification is made in the MRM mode, quantitation of unknown samples is normally carried out in the SIM mode.


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Geno/Grinder®To further reduce the clean-up steps, a new piece of technology was evaluated, called the 2000 Geno/Grinder® (SPEX SamplePrep, Metuchen, NJ). The Geno/Grinder is a laboratory mill and tissue homogenizer specifically designed for vigorous vertical shaking of deep-well titer plates. It was originally designed to prepare plant tissue for extractions of nucleic acids, proteins, and other constituents by shaking the tissue, steel balls and a buffering agent together in each well of a titer plate. The application of this tool can be expanded to include microorganisms when small silica beads are used instead of steel grinding balls. Microbes can also be disrupted in standard 96-well titer plates as opposed to deep-well plates. Sample material that can be prepared includes bacteria, yeasts, molds, seeds, stems, roots, leaves, and certain animal tissue. Due to the strength of the vertical shaking motion of the equipment, many seeds and other forms of plant tissue can also be pulverized dry in plates with the help of one or two grinding balls per well. Another benefit of this technology is that additional time savings are achieved because the sample vials are disposable, which means the laborious process of cleaning the homogenizer probes is eliminated.

Sample Clean-Up Using the Geno/GrinderFor the leaves of watery crops like tomatoes and potatoes, 10 g of sample was extracted twice with 20 mL of a 90/10 mixture of acetonitrile/water solution, using strong anion exchange (SAX) and the solid phase extraction (SPE) process described earlier. To ensure complete extraction of the compounds from the plant material, steel balls were added to the samples to pulverize the plant material. The samples were than extracted by shaking at high speeds in a Geno/Grinder at approximately 700-1200 cycles per minute, depending on the sample. The sample extract was brought to 50.0 mL by adding acetonitrile. Approximately 1 mL of the extract was filtered and 100 μL of sample diluted to 1.0 mL in an HPLC vial with mobile phase, and analyzed directly by LC/MS/MS. Using the Geno/Grinder has cut down the number of clean-up steps to 3, compared to 20 with the AMR 3705-95 method and 10 with the DuPont 13753 method.

ExperimentalTo evaluate the efficiency of the sample prep and clean-up stage described above, cymoxanil, the active ingredient in many of the fungicides, and its various metabolites, were determined in a number of wet and dry crop/plant materials. All samples generated were analyzed using the API 5000 LC/MS/MS (Applied Biosystems, Foster City, CA).

This instrument is coupled with an HPLC system to first separate the biological species of interest, based on their elution times off a column. The eluent is then introduced into the mass spectrometer for ionization to identify and quantitate the species using triple quadrupole MS technology. The principle of this technology is based on confirmation of a particular molecular ion by the generation of its daughter species using collisionally-induced dissociation or fragmentation (4). This technique, commonly known as multiple reaction monitoring (MRM), uses two resolving quadrupole mass filters, separated by another


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A 0.01 ppm cymoxanil LOQ Fortification Sample (DuPont - 13753) and control (blank) in the SIM mode is shown in Figure 3. It can be seen very clearly that there is a significant peak eluted at about 15 minutes, which corresponds to the cymoxanil. Confirmation of the compound is then made by measuring the intensities of the fragmented daughter ions in the MS/MS mode, by comparing them to known ratios in a data base of pesticides. The MS/MS spectrum (60- 300 Daltons) of cymoxanil in MRM mode is shown in Figure 4, while the resulting calibration curve for 0.2, 0.5, 1.0, 5.0 and 10.0 μg/L is shown in Figure 5.


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Figure 4. The MRM spectrum for cymoxanil showing the fragmented ions

Figure 3. Elution of cymoxanil using the LC/MS mode

Instrument MethodologyReversed-phase liquid chromatography was used to separate cymoxanil and its metabolites from the rest of the extractants. The LC/MS/MS instrument was operated in single ion monitoring MS negative ion mode for quantitative analysis. Peak area was used for quantitation. For confirmation of the presence of the analyte in unknown samples, the relative intensities of the fragmented ions were measured using the MRM mode. A brief summary of the HPLC and LC/MS/MS conditions are shown in Table 1.

HPLC System: Agilent 1100 SeriesHPLC Settings:

Injection Volume: 40 μL

LC Column: Agilent Zorbax® XDB-C8 (15.0 cm x 4.6 mm i.d., 5 μ)

Column Temperature: 30°C

Mobile Phases: 0.01 M Acetic Acid and Acetonitrile

LC/MS/MS System: Applied Biosystems API 5000LC/MS/MS Settings:

Ionization Source Electrospray

Polarity: Negative

Mode: Selected Ion Monitoring (SIM) and Multiple Reaction Monitoring (MRM)

Monitoring Mass/Charge: 197 Daltons (Cymoxanil, M-1)

Elution Timeframe: 0-17 min

Table 1. Brief Summary of the HPLC and LC/MS/MS instrumental conditions used for this study


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ResultsThe LC/MS separation of the experimental new insecticide under investigation and its metabolites (A-G) are shown in Figure 6.

Figure 6. The LC/MS separation of the experimental compound (DuP-1) and its metabolites (A-G)

The MRM (1-298 Daltons) of a 0.2 ppm standard of the metabolites D and E together with a tomato sample spiked at the LOQ is shown in Figure 7. Table 3 compares the Geno/Grinder sample preparation and clean-up method with a traditional homogenizer (Tissuemizer® probe) sample preparation using Dupont-13753 methodology described earlier. It shows quantitative data (ppm in sample) for the experimental new compound and one of its metabolites (D), for various crop samples. This methodology was then used to analyze a variety of wet, dry, oily and acidic crops for the active ingredient together with its metabolites. Table 4 shows spike recovery and RSD data for one of the watery crops (tomato), showing that DuP-1 and all its metabolites are within the guidelines stated in EPA OPPTS 860-1360 Multiresidue Test Method for testing of pesticides and toxic substances, which states that test data must have average recoveries between 70-120% with a precision of less than 15% RSD. Although they will not be presented here, the study also looked at other crops including limes (acidic crop), almonds (oily crop) and wheat straw (dry crop) and achieved similar spike recoveries and precision values.

Table 2 shows % recovery data and RSD for a 0.05 ppm and 0.50 ppm reference samples.

Reference AMR-3705-95 DuPont-13753 Geno/GrinderValue (% Recovery) (% Recovery) (% Recovery)

0.05 ppm (LOQ) 84 ± 3.4 (n=4) 93 ± 3.7 (n=5) 78 ± 3.1 (n=6)

0.50 ppm (10xLOQ) 68.5 ± 1.5 (n=2) 89 ± 2.4 (n=5) 78 ± 6.9 (n=4)

Table 2: % Recovery data and RSD for a 0.05 ppm (LOQ) and 0.50 ppm (10xLOQ) reference samples

Sample PreparationOnce the methodology was optimized for the separation and quantitation of cymoxanil, the following Geno/Grinder sample preparation and clean-up procedures were used for comparison purposes with traditional sample preparation methods. The basis of this method was then used to investigate an experimental new insecticide (DuP-1) and to identify and quantitate its active ingredient and metabolites in various wet/dry plant and crop materials

• Extract twice with Acetonitrile/H20 in Geno/Grinder for 2 minutes at 1200 cycles/min• Dilute 1ml of extract to 5ml with H20 (adjust to pH 2.5)• Filter through SAX SPE and collect extract• Dilute extract to 10.0ml• Filter through PTFE membrane into an LC vial• Analyze by LC/MS/MS


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Figure 5. Calibration curve for 0.2, 0.5, 1.0, 5.0 and 10.0 µg/L cymoxanil


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Table 4. Spike recovery and precision data for tomatoes showing that DuP-1 and all its metabolites are within the guidelines stated in EPA OPPTS 860-1360 Multiresidue Test Method

DuP-1 Met A Met B Met C Met D Met E Met F Met G

% Recovery & RSD of LOQ Sample 78+ 4.7 93+ 4.6 101+6.1 100+9.5 98+ 1.8 97+ 7.5 113+7.0 107+6.6

% Recovery & RSD of 10xLOQ Sample 80+ 5.8 82+ 6.8 83+ 8.0 85+ 4.8 78+ 8.1 76+ 4.5 82+ 6.9 77+ 9.6

860-1360 “Multiresidue Test Method for Pesticides and Toxic Substances”

ConclusionIt has been shown that the extraction of pesticide residues with the Geno/Grinder and identification and quantitation of its active ingredients and metabolites using LC/MS/MS, provides an extremely efficient, rugged and high throughput analytical method. Compared to the traditional sample preparation, clean-up and detection methods, sample throughput was increased by a factor of 3, from 8 samples to 24 samples per day.

Literature Cited1) Environmental Protection Agency, Office of Prevention, Pesticides and Toxic

Substances (OPPTS) Method

2) DuPont Report No. AMR 3705-95: “Analytical Method for the Determination of Famoxadone and Cymoxanil Residues in Various Matrices.”

3) DuPont Report No. 13753: “Analytical Method for the Determination of Cymoxanil and its Metabolites in Leafy Vegetables Using LC/MS”

4) B. A. Thomson, D.J. Douglas, J.J. Corr, J. W. Hager, C.A. Joliffe. Improved collisionally activated dissociation efficiency and mass resolution on a triple quadrupole mass spectrometer system. Analytical Chemistry, 67, 1696-1704 (1995)

Figure 7. The MRM of a 0.2 ppm standard of the metabolites D and E (top) together with a tomato sample spiked at the LOQ (bottom). The transition is shown at a molecular weight of 1-298 Daltons.

Table 3. Comparison between the Geno/Grinder and DuPont-13753 sample preparation methods for a variety of wet and dry crop samples

Crop DuPont - 13753 Geno/Grinder DuPont - 13753 Geno/Grinder Method Method Method Method

DuP-1 (ppm) Metabolite D (ppm)

Alfalfa 14.4 15.7 0.15 0.14

Corn Stover 0.003 0.002 N/D N/D

Cucumber 0.002 0.0015 N/D N/D

Green Onion 0.012 0.022 N/D N/D

White Straw 0.009 0.0075 0.003 0.002


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Clean UpAfter mixing, all samples from Sets A and B were centrifuged at 3500 rpm for 3 min. An aliquot (12-13 ml) of the supernatant liquid was taken from each sample and transferred to a clean 15 ml centrifuge tube. To each sample was added Primary Secondary Amine (PSA, 25 mg/ml sample), Graphitized Carbon Black (GCB, 5 mg/ml sample), and anhydrous Magnesium Sulfate (75 mg/ml sample) and the tubes were capped. Sample Set A was shaken by hand for 30 sec. Sample Set B was shaken on the Geno/Grinder for 30 sec. at 1500 spm. Following this treatment, both sample sets were visibly lighter in color.

All samples were centrifuged at 3200 rpm for 30 sec. The supernatant liquid was again removed from each sample and transferred to a clean 15 ml centrifuge tube. Samples were evaporated to near dryness using a slow stream of nitrogen gas and gentle heating. The total volume of each sample was brought to 1 ml in toluene and an internal standard mix (SPEX CertiPrep: CLPS-190) was added. The samples were analyzed by GC-MS using a SIM method.

GC-MS ConditionsAn HP 5890 gas chromatograph coupled to a 5972 mass selective detector was used for the analysis of the samples. The GC-MS was equipped with an HP 7673 GC/SFC injector. The analytes were separated on a HP-5 capillary column (3.0 m x 0.25 mm x 0.25 µm). The GC oven temperature program was set to an initial temperature of 70º C for one minute, and raised to 230º C at 20º C/min with a total run time of 15.0 minutes.

The MS was operated in electron impact ionization in the SIM mode a scan range of 35-450 m/z. The GC-MS interface and MS source were both 250º C The injected volume of sample extract was 2 µL with splitless injection.

Chemstation B.02.05 and Enviroquant G1701 BA Ver. B.01.00 were used for the data collection and analysis of samples.

Results and DiscussionThe results are shown in Table 1 and graphically in Figure 1. For all three matrices, the measured pesticide concentration was greater when the Geno/Grinder was used to shake the samples. The increase in recovery was similar for apple and strawberry, 18% and 20 % respectively. Both of these fruits contained the pesticide Flutianil. For pepper, recovery of the pesticide Etoxazole increased 35%. This indicates that use of the Geno/Grinder improves pesticide extraction from the fruit or vegetable matrix. At a setting of 1500 strokes/min., samples are shaken in a complete up and down motion 1500 times during a one minute run. In contrast, a lab worker can shake a sample approximately 200 times in one minute. Due to the more rapid and vigorous agitation using the Geno/Grinder, mixing should be more thorough than can be achieved manually and it is not surprising that extraction is improved over the manual method.

APPLICATION NOTE SP023: Comparison of Pesticide Extraction in Agricultural ProductsAPPLICATION: QuEChERS/Pesticide Extraction

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Comparison of Pesticide Extraction in Agricultural Products Using a Manual Shaking Method and Mechanical Mixing with the Geno/GrinderWith kind permission of: Matt Hengel, University of California, Davis, Lea Anderson-Smith, SPEX SamplePrep, Patricia Atkins, SPEX CertiPrep

ABSTRACTSince its introduction in 2003 by Anastassiades and Lehotay et al, the QuEChERS method1 (quick, easy, cheap, effective, rugged, safe) has proven to be effective and convenient for analysis of multiple pesticides in agricultural products. Increasing concern over the health effects of residual pesticides on fruits and vegetables has led to increased testing of these products to determine the levels of pesticides on produce when it goes to market. The QuEChERS method has allowed analysts to process a greater number of samples in a shorter period of time than with previous methods.

In this study, samples of strawberry, apple, and green pepper were prepared for analysis using both the standard QuEChERS method and a modified method involving mechanical mixing using the Geno/Grinder. The Geno/Grinder is a mechanical disrupter that grinds and mixes materials using a rapid vertical motion. GC-MS results of samples prepared using both methods were compared.

Experimental Sample PreparationMason jars of frozen, homogenized strawberry, apple, and green pepper (a blend of bell and non-bell) were obtained from UC Davis (IR-4 Project). Each fruit or vegetable contained a known pesticide, applied to the crop in the field; strawberry and apple contained Flutianil, and green pepper contained Etoxazole. The jars were thawed at room temperature prior to measuring out 15 g samples into 50 ml centrifuge tubes.

To each tube was then added 15 ml Acetonitrile containing 1% Acetic Acid. To the strawberry and apple samples, 6.0 g anhydrous Magnesium Sulfate and 1.5 g Sodium Acetate were added. To the green pepper samples, 6.0 g anhydrous Magnesium Sulfate and 1.0 g Sodium Chloride were added. All sample tubes were capped and the samples of each fruit or vegetable were divided into two sets, A and B. Set A was shaken manually, while Set B was shaken mechanically using the Geno/Grinder. Each sample set consisted of 5-7 samples.

Method 1 – Manual MixingSample Set A (from each material: strawberry, apple, and green pepper) was shaken by hand for 1 minute.

Method 2 – Mechanical MixingTwo angle cut ceramic cylinders (3/8” x 7/8”) were added to each tube in Sample Set B. The tubes were capped and shaken on the Geno/Grinder at 1500 strokes/min (spm) for 1 minute.


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Analysis of Pesticides in Fruit and Vegetable Products using a Standard QuEChERS Method and a Modified Method Involving the Geno/Grinder.With kind permission of: Lea Anderson-Smith, SPEX SamplePrep, Patricia Atkins, SPEX CertiPrep

IntroductionPesticide residues in agricultural food sources are widely considered to cause adverse health effects when consumed by humans. In particular, much of the produce sold in the U.S. is imported and concern over pesticide levels in these fruits and vegetables in comparison to those grown domestically has resulted in increased testing for pesticide residues.

In 2003, the QuEChERS method for pesticide analysis was introduced by Anastassiades and Lehotay et al. QuEChERS is an acronym for Quick, Easy, Cheap, Effective, Rugged, and Safe and allows for analysis of multiple pesticides, while offering faster and easier handling over previous methods. The QuEChERS method is now widely used and has been adopted by the AOAC as method 2007.01 “Determination of Pesticide Residues in Foods by Acetonitrile Extraction and Partitioning with Magnesium Sulfate.” In Europe, the official method is EN 15662.

For a typical analysis, 10-15 g samples of chopped and homogenized agricultural produce are placed in 50 ml centrifuge tubes and an extracting solvent, such as acetonitrile, and anhydrous magnesium sulfate and sodium acetate or sodium chloride salts are added. The tubes are capped and shaken by hand for one minute to mix the contents and extract the pesticide into the solvent. The samples are then treated with clean up materials, concentrated, and analyzed by GC/MS or LC/MS.

In this study, the Geno/Grinder was employed to homogenize the fruit/vegetable samples and to mix the produce rapidly and thoroughly with the salts and solvent in an effort to improve the extraction step. The goal of the study was to determine whether the use of the Geno/Grinder during the extraction step would increase pesticide recovery over the traditional, manual QuEChERS method.

ExperimentalThree fruit and vegetable matrices were chosen for the purpose of evaluating materials with differing density and toughness: 1) strawberries – soft; 2) apples – dense and tough; and 3) celery – fibrous. Strawberries are quite soft and preliminary tests verified that they could easily be ground to a mushy, liquefied substance using the Geno/Grinder (grinding method described below). Apples are tougher and denser and, not surprisingly, a longer grinding time was required to effectively mash the fruit to an applesauce-like consistency. While celery is not as dense as apple, it is fibrous and tough and therefore difficult to grind effectively.

Fresh strawberries, apples (Granny Smith), and celery were purchased from a local supermarket and cut

APPLICATION NOTE SP024: Pesticide Anaylsis: Standard QuEChERS vs Modified MethodAPPLICATION: QuEChERS / Pesticide Extraction

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The addition of two ceramic grinding cylinders to each tube shaken on the Geno/Grinder also may have aided the extraction process. The ceramic cylinders mash and grind the fruit or vegetable matrix further during the shaking process, thereby increasing surface area and allowing a more complete extraction of pesticide from the matrix. In fact, the matrices in the samples shaken on Geno/Grinder visibly appeared more thoroughly ground than samples shaken by hand.

Table 1. Comparision of GC-MS Results for Samples Prepared Manually and Using the Geno/Grinder

Manual Geno/Grinder

Matrix Conc. (ppb) Std. Dev. RSD (%) Conc. (ppb) Std. Dev. RSD (%) Increase using Geno/Grinder (%)

Strawberry 20 1.0 5 24 1.7 7 20

Apple 9 0.8 9 11 1.4 12 18

Pepper 17 2.5 15 23 2.4 10 35

An additional benefit of using the Geno/Grinder for sample preparation is elimination of variation in shaking technique. Samples are held vertically in a foam support and every sample within a run is subjected to the same shaking conditions. Also, since the operating rate and run time can be set on the Geno/Grinder, identical conditions from run to run are ensured. However, when samples are shaken manually, shaking conditions can vary and this can affect how well the samples are mixed. As a worker becomes tired from shaking multiple samples, he/she will likely slow down and later samples may not be as well shaken as earlier one. If two are more people are preparing samples, there is also the possibility that the workers will not shake the samples in the same manner. Again, this can affect the effectiveness of the extraction.

Finally, use of the Geno/Grinder allows for greater sample throughput. While a lab worker can shake 2-4 samples (50 ml tubes) simultaneously by hand, the Geno/Grinder can accommodate 16. Thus, more samples can be prepared in a given amount of time.

1. QuEChERS method developed by the USDA Eastern Regional Research Center. M. Anastassiades, SIJ. Lehotay, D. Stajnbaher, F.J. Schenck, J.AOAC International 86, p. 412-431 (2003).


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fruits and vegetables tended to become packed down in the bottom of the tube and effective grinding was difficult to achieve. Alternatively, addition of the full 15 ml of solvent prior to grinding provided too much fluid and the grinding media were not able to make sufficient contact with the produce to achieve effective grinding. Through experimentation, it was determined that addition of 5 ml of solvent was optimal for effective grinding of a 15 g sample.

To each tube was then added 6 g anhydrous magnesium sulfate, 1.5 g anhydrous sodium acetate, and the remaining 10 ml acetonitrile (1% glacial acetic acid). The tubes were recapped, placed back on the Geno/Grinder and shaken for 1 min. at 1500 rpm. The liquid in the strawberry tubes was observed to be pink in color, the apple extract was pale yellow, and the celery extract a very saturated green color. All material was well mixed and free-flowing.

The samples were then centrifuged at 3500 rpm for 3 min. The supernatant liquid was removed from each tube, measured, and divided into two equal samples (5 ppm max. pesticide after this step) and transferred into 15 centrifuge tubes.

PSA (25 mg x Vol (ml) supernatant) and GCB (5 mg x Vol (ml) supernatant) were added to each tube. The sample tubes were capped, shaken on the Geno/Grinder for 30 sec. at 1500 rpm, then centrifuged at 3200 rpm for 1 min.

Modified QuEChERS Method using the Geno/Grinder – All-In-OneAdditional samples of spiked strawberry were handled in a second manner. In an effort to determine whether the pre-homogenization step can be eliminated for soft fruits, 6 g anhydrous magnesium sulfate, 1.5 g anhydrous sodium acetate, and 15 ml acetonitrile (1% glacial acetic acid) were added to tubes containing 15.1 g of chunked strawberry. The tubes were capped and clamped on the Geno/Grinder. After 2 min. of grinding at 1500 rpm the strawberry was fairly well ground, but some visible chunks remained. The samples were ground an additional 2 min. (4 min. total), at which point the strawberry was very well ground and thoroughly mixed with the salts and solvent. The samples were centrifuged, the supernatant removed, and clean up was conducted as above, using the Geno/Grinder.

Attempts to combine the homogenization and extraction steps for apple and celery were less successful. When the salts were combined with small chunks of produce, a tube was filled to capacity and the produce and grinding media could not move freely. Thus, impact of the grinding media on the produce was impeded and only partial grinding was achieved.

Preparation of SamplesFollowing clean up and centrifugation, the supernatant was removed from each sample, transferred to a clean 15 ml tube (all methods), and concentrated down to near dryness (approximately 100 µL) by gently heating the tubes while passing a slow stream of nitrogen over the sample. Toluene was added to each sample to bring the total sample volume to 1 ml. In many cases a viscous drop of material settled to the


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into 1/4-1/2 inch-sized chunks, weighed out in 15.1 g quantities, and placed into 50 ml round-bottomed LDPE centrifuge tubes. Each sample was spiked with 250 µl of a 40 µg/ml solution of CAL-CARB-13 (a mix of 13 common pesticides available from SPEX CertiPrep) in dichloromethane (10 ppm per sample). The pesticide mix was introduced into each tube by syringe, taking care to control the flow of pesticide solution onto the fruit/vegetable sample. The tubes were capped and gently shaken by hand for 15 sec. to ensure an even distribution of the pesticide solution over the entire sample. The samples were placed in a refrigerator at 4°C and stored overnight.

Traditional QuEChERS MethodThe spiked strawberry samples from four centrifuge tubes (each containing 15.1 g) were collectively transferred into a single-serve blender and homogenized to a smooth consistency. Samples of 15.1 g were re-measured from this mixture and transferred back into the same centrifuge tubes. Care was taken to transfer out liquid as well as solid material. Apple and celery samples were spiked and homogenized in the same manner as the strawberry.

To each tube of homogenized produce was added 6.0 g anhydrous magnesium sulfate, 1.5 g anhydrous sodium acetate, and 15 ml acetonitrile containing 1% glacial acetic acid. The tubes were capped and shaken by hand for 1 min. The liquid in the strawberry tubes was observed to be pink in color, the apple extract was pale yellow, and the celery extract a very saturated green color.

All tubes were then centrifuged at 3500 rpm for 3 min. The supernatant liquid was removed, measured, and divided into two equal portions (5 ppm maximum pesticide per sample after this step) and transferred into 15 ml centrifuge tubes.

Primary secondary amine (PSA) (25 mg x Vol (ml) supernatant) and graphitized carbon black (GCB) (5 mg x Vol (ml) supernatant) were added to each tube. The sample tubes were capped and shaken by hand for 30 sec., then centrifuged at 3200 rpm for 1 min.

Modified QuEChERS Method using the Geno/GrinderTo centrifuge tubes containing spiked produce samples were added 3 ceramic grinding cylinders (3/8” x 7/8” angle cut) and 5 ml acetonitrile containing 1% glacial acetic acid. The tubes were capped and placed into a tube holder, clamped in place on the Geno/Grinder, and ground at 1500 rpm. After grinding for 2 min., the strawberry samples were well ground to a pulpy, liquid consistency. Since apple and celery are tougher materials, 6 min. grind times at 1500 rpm were required to achieve a well ground, mushy consistency.

During preliminary test runs, it was found that addition of a small amount (5 ml) of solvent improved the grinding process by aiding the movement of the produce in the tube. Without solvent, the harder


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Table 1 – Recovery of Pesticides in Produce Samples (ppm)

Pesticide Strawberry Apple Celery

Geno/Grinder All in one QuEChERS Geno/Grinder QuEChERS Geno/Grinder QuEChERS

Carbofuran 2.33 2.2 0.8 2.4 1.2 1.0 ND*

Carbaryl 1.8 1.6 ND 1.8 0.4 ND ND

Diphenylamine 1.8 2.2 ND 6.2 3.8 0.9 ND

Chlorpropham 2.7 3.3 0.3 3.0 1.1 1.5 0.1

Dichloran 0.7 0.4 ND 0.4 ND 0.2 ND

Chlorothalonil 0.7 ND ND 1.2 ND ND ND

Pirimicarb 1.8 1.4 ND 1.4 ND 0.1 ND

Vinclozolin 2.5 2.9 0.1 2.8 1.1 2.3 ND

Metalaxyl 3.1 3.2 1.3 3.0 1.8 2.3 1.5

Parathion 2.9 3.4 1.3 3.0 1.9 3.2 1.4

Systhane 2.8 3.1 1.0 3.0 1.6 2.3 1.1

Azinphos-Methyl 2.5 2.7 1.9 3.1 2.9 3.2 2.4

*ND = Not Detected

Figure 1 – Recovery of Pesticides in Strawberry Samples


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bottom of the centrifuge tube after addition of toluene. The material appeared to be soluble in aqueous solvents, but not in non-polar solvents and had a syrupy consistency.

The toluene solution was removed by syringe and injected into GC sample vials, while the residue material was left in the tube. For strawberry the residue was red for the samples that had been prepared using the Geno/Grinder and yellow for samples prepared using the standard QuEChERS method. In the case of apple, the residue was golden for Geno/Grinder samples and brown for standard QuEChERS samples. For celery, a yellow residue remained for Geno/Grinder samples, while for standard QuEChERS samples a pale yellow drop was observed.

Sample AnalysisSamples were analyzed using an HP 5890-GC with a CV-5 capillary column and a 5972-MSD detector. Scan range was 35-450 m/z with signal-to-noise of 3:1. Injected sample size was 1 µl.

Results and DiscussionAt the beginning of the study, each sample was spiked with 250 µl of a solution of 13 pesticides, each present in a concentration of 40 ppm/ml. Thus, 10 ppm of each pesticide was introduced into each produce sample. After extraction and centrifugation, the supernatant was removed and divided into two equal replicates to continue the study. Therefore, each sample was divided into two and the maximum concentration of each pesticide in a sample at this point was 5 ppm.

Recovery results for all three produce matrices are shown in Table 1 as total concentration recovered in ppm. Results for strawberry are shown graphically in Figure 1. The blue bars represent the samples that were prepared by manually shaking homogenized samples with the salts and acetonitrile. The red bars represent the samples that were homogenized using the Geno/Grinder prior to the addition of the salts. Finally, the green bars (All in one) show the results for the samples that were ground and mixed with the salts and acetonitrile simultaneously (no pre-homogenization).

For all pesticides detected, significantly higher recoveries were obtained for the samples prepared using the Geno/Grinder than for those prepared by hand. Interestingly, the samples ground simultaneously with the salts and solvent (All in one)performed as well, or in some cases better, than the samples that were ground on the Geno/Grinder before addition of the salts. This indicates that this method is viable for soft produce matrices.

The poorest recoveries were obtained for Dichloran and Chorothalonil. This was the case for all matrices evaluated. In fact, for celery Chlorothalonil was not detected, even when the Geno/Grinder was used. Because the method used in this study was not optimized for a particular type of pesticide, it is not surprising that some pesticides were not stable under the conditions used.


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Figure 3 - Recovery of Pesticides in Celery Samples

It is clear from the preceding graphs that use of the Geno/Grinder significantly increased pesticide recovery in this study. Since the Geno/Grinder shakes the samples so much more vigorously and rapidly than can be accomplished by hand, it is not surprising that extraction was more effective. During a 1 minute run at a setting of 1500 rpm, the Geno/Grinder shakes a tube in a complete up and down cycle 1500 times, whereas a human can shake a tube about 200 times in 1 minute. In addition, the sample tubes run on the Geno/Grinder each contained 3 ceramic, angle-cut grinding cylinders. These aid in mixing the sample thoroughly with the solvent and salts, but also grind the produce, ensuring a thorough extraction of the pesticide. Thus, for soft produce matrices, it may be possible to eliminate the pre-homogenization step, thereby further reducing sample preparation time.

In addition to improved pesticide recovery, the use of the Geno/Grinder increases throughput since a greater number of samples can be homogenized and extracted at one time. In this study, twelve 50 ml tubes were shaken simultaneously on the Geno/Grinder, whereas a maximum of four tubes were shaken by hand at one time. During the clean-up step, the Geno/Grinder accommodated twenty-four 15 ml tubes, while a maximum of six were shaken manually.

ConclusionProduce samples of varying density and toughness were evaluated and in all cases significantly greater pesticide recovery was obtained for samples prepared using the Geno/Grinder than for samples shaken manually.

In addition, since run time and operating rate are automatically controlled by the Geno/Grinder, all samples within a run and from run to run are shaken in a consistent manner, eliminating variability. The Geno/Grinder can also shake up to sixteen 50 ml centrifuge tubes in one run, thus increasing sample


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Figure 2 shows the recovery results for apple. Again, the Geno/Grinder method outperformed the standard manual method. Greater recoveries were obtained for all detected pesticides when the Geno/Grinder was used to grind the apple and mix the resulting pulpy mass with the salts and acetonitrile, with the exception of Azinphos-Methyl where the recovery for the two methods was similar. For most pesticides, the recoveries were similar to those obtained from the strawberry samples. However, the Diphenlyamine results from the Geno/Grinder samples gave a recovery of 6.2 ppm, which is greater than the 5 ppm that was introduced to the sample. The recovery for this pesticide using the manual method (3.8 ppm) was also higher than for the other pesticides for these same samples. Diphenylamine is commonly used in crop protection for apples and the high concentration for this pesticide is likely a result of its presence on the apple before the addition of the pesticide spike. Nevertheless, the results for the Geno/Grinder samples were significantly higher than for the standard QuEChERS samples.

Figure 2 - Recovery of Pesticides in Apple Samples

Results for celery are shown in Figure 3. As with strawberry and apple, the recoveries for the Geno/Grinder samples are much higher than for the manually prepared samples. For all three matrices evaluated, the Geno/Grinder extracted some pesticides that were not detected in the samples prepared by hand. Particularly noteworthy in the case of celery is Vinclozolin; the recovery is on par with Metalxyl and Systhane for samples prepared using the Geno/Grinder, but was completely undetected in the standard QuCEhERS samples.


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Quick DNA Extraction from Rice Seed (Wet)With kind permission of RiceTec Inc, Alvin TX

ABSTRACTDNA extractions can be a very time consuming and tedious process. Finding a quick method in which DNA could be extracted and used for PCR is essential. Described below is a quick “dirty” method that produces a high enough concentration of DNA that can be used for PCR.

Sample ExtractionSamples are prepared using a 96 well 1ml assay block. Dispense one 5/32” stainless steel bead (4mm) into each well using the Grinding ball dispenser (SPEX SamplePrep cat. # 2100). Next, add one seed to each well. Dispense extraction buffer into each well and securely cap each well. After the samples have been capped, grind them in the Geno/Grinder2000 (SPEX Sample Prep 2000-115) at 500 strokes/minute for two minutes. Centrifuge for 1 min to bring all liquid to the bottom of the assay block. Incubate the samples in about 1” of water at 95ºC for 20 minutes then place them on ice for approximately 10 minutes or until samples are cool to the touch. Centrifuge again for 1 minute. Add neutralizing extraction buffer and seal the assay block with sealing film. Centrifuge the samples for 10 minutes at 3000rpm. Transfer 300µL of the supernatant to a clean 96 well plate. DNA can be further purified with clean-up kits available on the market.

YieldsThe total concentrations yielded from the samples range from 3-7ng/µL in a final volume of 200µL with purities of 1.5-1.8.

ConclusionThe method described above is sufficient for PCR and it takes less time than the standard chloroform extraction. Total time ranges from one to two hours.

APPLICATION NOTE SP025: Lysing / HomogenizationAPPLICATION: DNA Extraction from Rice Seeds

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throughput. This not only reduces time spent on sample preparation, but also reduces fatigue for laboratory workers.

Finally, produce samples can be ground in 50 ml centrifuge tubes using the Geno/Grinder, possibly eliminating the need to pre-homogenize the produce. Soft fruits, such as strawberries, can be ground in the presence of the salts and extracting solvent used in the QuEChERS method with no negative impact on pesticide recovery.


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How To Subject: Grinding Bone in the Freezer/Mill

HOW TO Guidelines: Cryogenic Grinding of Bone in SPEX SamplePrep Freezer/Mills

Any plant or animal tissue can be ground in a SPEX Freezer/Mill, and this includes all bone: fresh or aged, wet or dry. Freezer/Mill grinding vials are immersed in liquid nitrogen (LN) before and during grinding, first to make the sample brittle enough to grind, and then to keep it brittle during the actual grinding. The actual grinding is done by a rod-shaped steel impactor that is magnetically shuttled back and forth between two steel end plugs.

Success in Freezer/Mill cryo-grinding depends on two main things: The sample must be cold enough to be brittle. The impactor must be able to move freely back and forth.

The quantity of sample per load and the size of the sample pieces can heavily affect the grinding efficiency of the mill, particularly with bone, which can be very tough. A simple test is to load the vial, then shake it back and forth to make sure the impactor can move from end to end of the vial. If the sample pieces are too large or there is too much sample in the vial, the impactor can jam, or not move far enough to grind effectively. In extreme cases, if the impactor is deflected sideways or wedged against the polycarbonate cylinder by a large piece of bone, the cylinder can crack or break.

To grind bone in the standard 6751 vial, we recommend cutting it before grinding into pieces approximately 5 mm in diameter, or smaller. The quantity to be ground in a 6751 Vial can vary from less than half a gram up to four or five grams, depending on the toughness of the sample and the size of the pieces. The optimum sample size and quantity for your samples should be determined empirically. It may be possible to grind one or two large pieces of bone at a time – for example, a 2-gram chunk in one end of the vial – but the uniformity of the results will be better and there will be much less risk of cracking a cylinder if the bone is cut into smaller pieces first.

For bone in the 6751 vial we recommend at least 10 minutes of Pre-Cool time. Depending on the quantity and size of the bone pieces, grinding can take anywhere from 1 minute to 10 minutes. Grinding times longer than 2 minutes should be split into 2- minute grinding periods separated by 2 minutes of cooling. Grinding can be speeded up by increasing the grinding rate from the factory setting of 10 to as much as 14 or 15.

If the 6751C polycarbonate center cylinders persist in cracking or breaking, we recommend the use of the 6752 steel center cylinders. These do not form a tight seal with the end plugs when chilled (as do the polycarbonate cylinders), and the joints between the end plugs and steel cylinder should be sealed by a piece of Teflon plumber’s tape wrapped around the shank of each end plug.

Preparation of Domestic Waste for Elemental AnalysisWith kind permission of IBU-tec GmbH & Co. KG

Domestic waste presents a very inhomogeneous mix made up of very different substances. Apart from polymer synthetics and organic substances like cork, wood, and paper there may be inorganic materials and metals present. In order to use this domestic waste in a useful way it is burned in waste incinerating plants to gain heat combustion.

Non-Uniform CompositionAs well as the elemental analysis the G.H.V. and N.H.V. (gross and net heat value) is of interest. To this end the waste has to be ground and homogenized as well as possible. The variable composition does not allow the use of a traditional ball mills or swing mills. Cutting mills could possibly grind these samples but not to the particle size necessary for organic elemental analysis.

Brittleness Through CoolingThe Freezer/Mill (a cryogenic mill) is effective where these grinding processes fail. Flexible materials like plastics and biological samples embrittle at liquid nitrogen temperatures. The sample is hermetically sealed in the grinding vial and immersed in liquid nitrogen, so cross contamination is eliminated. As the Freezer/Mill is driven magnetically there is no wear and tear on moving parts. For the best possible grinding results the sample has to be cooled to the optimum. To this end the roughly pre-ground sample can be pre-cooled in the grinding vial or poured into it already cooled. After 20 minutes of pre-cooling the sample was ground for three minutes, followed by one minute of ‘inter-cooling’ to ensure good brittleness and then ground further. This cycle was repeated three times.

ResultsCryogenic grinding with the Freezer/Mill showed good results with the homogenization of domestic waste. The obtained powders displayed good particle size and homogeneity, suitable for elemental analysis. Following the results of organic elemental analysis the G.H.V. and N.H.V. of the waste sample can be calculated and thus ascertained if and how much fuel has to be added for optimum combustion in the incinerating plants. With sulfur analysis information regarding a possible environmental impact can be gained. Finally, ash analysis after incineration of domestic waste samples gives information about the remaining residues that have to be disposed of.

Application Note SP002: Ginding/Homogenization :: APPLICATION: Domestic Waste

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RNA Extraction from Cartilage Tissue Using Cryogenic GrindingWith kind permission of ALTANA Pharma AG

The object of the study was to determine if there is a connection between the seriousness of arthritic diseases, especially of the knee joint and certain genes. To this end the expression profile of various genes in the tissue of healthy and diseased probands was compared. Total RNA was isolated from cartilage tissue and the activity of certain genes was analyzed via PCR or micro array analysis.

Human Cartilage TissueKnee cartilage tissue samples were taken from various patients as well as healthy probands, snap-frozen in liquid nitrogen and stored at -80°C. One to two grams of these samples were taken for RNA extraction and added frozen to the grinding container. With the multi-vial adapter option up to four samples can be homogenized simultaneously in the Freezer/Mill® 6850. By utilizing the precool function of the 6850, the samples were held at about -196°C for another two minutes. This assured sufficient cooling and therefore optimum brittleness.

523Grinding Procedure / HomogenizationFor technical reasons,—due to the large surface area and the small particle size the cartilage powder showed a tendency to thaw quickly, which should be absolutely avoided (RNA degradation)—the phenol-chloroform reagent (peqGold RNAPure™, peqLAB; Erlangen, Germany) used for further preparation had already been added to the samples (approx. 2 to 2.5ml). After adding the reagent the grinding vials were placed in upright position into liquid nitrogen until the buffer solidified. Then the cartilage samples and the impactor were added to the cooled vial. In order to assure that the impactor not be frozen onto the reagent, the Freezer/Mill was started briefly to check the movement by listening to the sound of the mill.

The grinding was done in three cycles of two minutes each at maximum frequency. Between grinding cycles samples were cooled for two minutes to achieve optimum brittleness again. Thus the whole procedure took 12 minutes. The resulting powder was transferred into Falcon tubes and charged with more peqGOLD reagent (approx. 20ml).

PreparationThe RNA was subsequently purified using a phenol-chloroform extraction method according to the producer’s protocol. In order to guarantee optimum RNA clean-up and quality the resulting liquid phase was applied onto RNeasy midi columns.

Application Note SP005: DNA/RNA Extraction :: APPLICATION: PCR

SP005HOW TO Guidelines HT001

The large 6801 vial can grind up to 40 or 50 grams of bone, and the size of the pieces is not important unless they are large enough to prevent the impactor from moving. However, the 6801 vial usually takes longer to grind a sample than the 6751 vial, and for a large sample we recommend 15 minutes of pre-cooling. As with the 6751 vial, grinding should be programmed for two minutes on and two minutes of cooling for as many cycles as necessary to grind the bone to the desired particle size range. The grinding rate can be left at the factory setting of 10 or increased until the impactor starts to “stutter;” the maximum effective rate for the 6801 vial to run smoothly is usually 12.

Grinding performance in the Freezer/Mill is affected by the quantity of sample, the size of the sample pieces, and grinding time. To improve performance, decrease the overall sample quantity, pre-cut the sample into smaller, more uniform pieces, or grind for additional cycles. 10 minutes of pre-cooling for the 6751 vial and 15 minutes of pre-cooling for the 6801 vial ought to be enough for bone, but for very large pieces of bone in the 6801 you may need to increase the pre-cooling time to 20 minutes.

Again, we recommend running the Freezer/Mill for 2 minutes per grinding cycle with 2 minutes of cooling between cycles, to let the sample and coil cool back down. It is possible to run the Freezer/Mill continuously, but as the coil heats up it becomes less efficient. Bone will probably remain “grindable” for many minutes of continuous grinding, but for difficult-to-grind samples like synthetic fibers and polymer sheets, it is probably better to keep them as cold as possible during grinding.

Generally speaking, all components of Freezer/Mill vials can be cleaned with hot water and detergent or soap, and decontaminated by dipping in a solution of 10% bleach or dilute mineral acids. After cleaning they should be dried. Polycarbonate cylinders can be damaged by excessive heat (as in autoclaving) or exposure to organic solvents including alcohol. Steel impactors and end plugs, and steel cylinders, may be sterilized by disinfectants, alcohol, autoclaving, etc., but should not be left in contact with water and aqueous fluids for any length of time, as magnetic steel can rust under these conditions.

For further advice about Freezer/Mill applications and maintenance, please call Customer Service at 1-800-LAB-SPEX x465, or 732-549-7144 x465. You can also contact us via email at [email protected].

How To Subject: Grinding Bone in the Freezer/Mill


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Preparation of Rice Husks for the Analysis of Their ContentsWith kind permission of IBU-tec GmbH & Co. KG

For a very large number of the global population (approximately 50%) rice is the primary foodstuff. People in Asia and Africa virtually exist only on rice. As a result of this there are incredibly large rice crop areas especially in Asia.

Health AspectHowever, almost everywhere the rice is eaten as white rice (polished), i.e. the bran and the germ are removed. It has been known for a long time that this can cause deficiency diseases, especially avitaminose (Beri-Beri). As well as the vitamins B1, B2, B3 and K the husks also contain important minerals (calcium, magnesium, selenium, zinc). Therefore the husks can be considered as a resource. For the purpose of content analysis the rice husks were ground and homogenized in the Freezer/Mill® 6850, using the large grinding vials.

Cool GrindingThe Freezer/Mill has a magnetically driven impactor. This impactor is inserted into the vial along with the sample to be ground, this assembly is then placed into the coil assembly within the mill, which is lowered into the liquid nitrogen bath. Grinding is accomplished by alternating the magnetic field in the coil assembly forcing the impactor into the end caps. The most important aspect for a successful grinding procedure is the sufficient cooling of the milling material to ensure brittleness of the sample. To this end the filled vial is inserted into the opening of the coil assembly and immersed into a liquid nitrogen bath. For brittleness the sample was left in the nitrogen bath for approximately 12 minutes. Subsequently the sample was ground for three minutes, then after waiting for one minute to renew good brittleness, it was ground again. All in all three of these cycles were performed, which corresponds to a grinding time of nine minutes. For this kind of grinding program approximately eight grams of starting material was used.

ResultsAll in all very good homogenizations of the rice husks were obtained. This was shown by the fineness of the resulting powder (analytical fineness) on the one hand and the analysis results which exhibited good reproducibility on the other hand.

OutlookThe Freezer/Mill has proved to be very suitable for milling and blending of slightly brittle yet flexible plant parts for analysis. With the Freezer/Mill 6850 this can be done in units of a few grams. The low temperature causes brittleness of the material and any generated heat, which would have impeded effective grinding, is avoided.

Application Note SP006: Milling/Blending (Homogenizing) :: APPLICATION: Plant Parts

SP006

Application Note SP005: DNA/RNA Extraction :: APPLICATION: PCR

After photometric examination of the RNA, the samples were analyzed either by quantitative PCR after reverse transcription or alternatively with the Affymetrix GeneChip® system.

The grinding vials were cleaned in several steps. First they were cleaned mechanically with tissue paper and 70% alcohol, then with soap suds. After rinsing with water they were again rinsed with 70% alcohol. This cleaning process should always be carried out promptly as the thawing phenolreagent corrodes the vials (the stainless steel accessories are not affected). Prior to the next use the whole grinding set was autoclaved.

Yield of cartilage RNA was between 2 and 20 μ/g RNA depending on the patient’s samples.

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B) Material Analysis: INOC-TestDepending on the stabilizer content, about 200 to 400mg of sample material and 10.0mg of AIBN (initiator) are weighed accurately into the reaction vessel. 10.0ml of Cumol is then added with a pipette. The vessel is connected to the oxidation apparatus (see fig.) The Cumol efficiently extracts the stabilizer contained in the pulverized fleece material permitting AIBN oxidation.

The INOC Test measures time dependent oxygen consumption during the reaction. From the data obtained, a diagram is generated enabling visual evaluation of the stabilizer content in the samples.1)

ConclusionThe 6750 Freezer/Mill efficiently reduces the particle size of samples that are difficult to grind without thermal degradation of the material, for example fleece materials to a particle size of less than 500μm. Sample preparation time is drastically reduced to less than 15 minutes due to the increased brittleness of the samples at these very low temperatures.

Apparatus:

Application Note SP007: Grinding/Homogenization :: APPLICATION: Polymer Yams & Fleece Materials

Water bath

Valve Capillaries

Pre-set level(constant pressure)

Flexible tubing

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Grinding Polymers for Qualitative and Quantitative AnalysisWith kind permission of the Federal Institute for Materials Research and Testing (BAM)

In the application discussed here, the 6750 Freezer/Mill® was used for grinding compact synthetic (pellets, webbing fabric, textiles, etc.) as well as polymer yarns, fleece material, filaments (polypropylene (PP), polyethylene or polyester). Grinding was required for accurate qualitative and quantitative analysis of additives contained in synthetic materials (e.g. antioxidants using HPLC and INOC-Test). The Freezer/Mill is ideal for this application, providing the required particle size reduction in a relatively short time period.

Example: PP Fleece MaterialsThe material is first cut into approximately 2mm squares. The grinding vials are then filled half way with sample; then ground as per the program below:

Pre-cooling : 15 min Run : 1.5 min Cycle cool : 2 min Cycles : 5 Rate : 10Hz

For the analysis discussed here, the method above was found to be the optimum grinding program for PP fleece materials. The yield is high (80% - 90%) and during the short grinding times the material does not warm enough to lose its brittleness or decompose. The number of grinding cycles is dependent on the quantity of sample placed in the vial and the desired final particle size. In order to exclude particles that are too coarse the ground fleece material can be sieved, for example passed through a 500μm sieve.

A) Sample Preparation for HPLC Analysis of AdditivesApproximately 2.0g of the ground material is weighed into a 125ml flask to an accuracy of 1mg and 40ml toluene is added. The sample is brought to a boil with constant stirring. After approximately 30 minutes the material has completely dissolved, the solution is than slowly cooled to 50°C. In order to precipitate the polymer, 40ml of methanol is added with further stirring. The mixture is then cooled to room temperature and filtered through a Nutsche (suction) filter. The filtrate is then passed through a 0.45μm filter. The stabilizers necessary for analysis are now in solution and ready for oxidation and HPLC separation.

Application Note SP007: Grinding/Homogenization :: APPLICATION: Polymer Yams & Fleece Materials

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Nafion®-Powder for the Production of Fuel Cell ElectrodesWith kind permission of DLR (German Aerospace Center) Stuttgart; Electrochemical & Electrical Engineering

The Freezer/Mill®, a cryogenic mill is useful where other grinding procedures fail. Flexible materials like synthetics and biological samples are embrittled at Liquid Nitrogen temperatures. The sample is hermetically sealed and cross contamination is impossible. Since the Freezer/Mill is driven magnetically there is no wear and tear on the movable parts.

Special PolymerNafion® a polymer similar to Teflon (perfluorosulfonate polymer), which has been used for over 30 years as a proton conducting membrane in fuel cells, serves as an electrolyte. Because of its properties it proved very suitable for this application. The DLR has now developed a method to apply the Nafion in powder form. For the production of this Nafion powder, the Freezer/Mill, a cryogenic mill is utilized. As a polymer Nafion is very flexible and soft which obviously makes milling difficult. The Nafion granules (5mm particle size) are reduced to approx. 10μm during grinding. The resulting powder is mixed with a catalyst and sprayed dry onto to the 25 - 175 μm thick membrane.

Fuel CellThe fuel cell generates electrical output through electro-chemical oxidization of hydrogen into a proton and an electron as well as oxygen reduction as counter reaction. In order to reduce the necessary activation energy for this reaction a catalyst has to be used. In the PEM-MC (Polymer- Electrolyte-Membrane-Fuel Cell) platinum is used as a catalyst. The direct methanol fuel cell needs ruthenium as a catalyst in addition. These polymer membranes are utilized in various fuel cells.

Result (Conclusion)Despite the special properties of Nafion, which as a polymer is very soft and flexible, it was possible to obtain a very fine powder after a combined grinding time of 20 minutes with the freezer/mill. No particle size determination was carried out, although because of the thickness of the applied layer the maximum particle size can be specified as less than 25μm.

Application Note SP009: Reduction/Homogenization :: APPLICATION: Polymers

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Structural Changes in Polymers by Means of Cryogenic GrindingWith kind permission of Prof. Dr. U. Köster, Dept. of Chemical Engineering , University of Dortmund

One goal in materials research is to develop new synthetic materials with characteristics that cannot be accomplished by the usual smelting process, as the polymers tend to separate during the heating process.

Pure PolymerTrials to introduce permanent changes in polymers by means of Cryogenic Mechanical Milling (CMM) with the Freezer/Mill® were started with two polymers; isotactic polypropylene (iPP) and syndiotactic polystyrol (sPS). After melting and re-crystallization, both cryogenically ground (i.e.at–196°C) polymers showed characteristics that differ from the original polymers. DSC (Differential Scanning Calorimetry) tests showed changed crystallization temperatures, which indicates reduced molecular weight or a change in molecular weight distribution. The longer the samples were subjected to cryogenic grinding the bigger the change.

Further experiments showed that the crystalline structure is increasingly destroyed during cryogenic grinding. Presumably CMM leads to chain fission and therefore to the formation of macro radicals. This can lead to cross-linking and an increased amorphic content in the partially crystallized polymers. Another explanation would be the formation of nano structures, small fragments of the original polymer compounds.

CompoundsAs well as the pure polymer results the behavior of cryogenically ground polymer compounds is, of course, very interesting. When mixing the two polymers iPP and sPS through melting (extrusion) separation occurs. Such polymer blends are dispersions of one part within the other. Some CMM sample tests suggest that re-combination of constitutive different macro radicals partly results in co-polymers. These co-polymers could act as phasing facilitators between the incompatible blending components of iPP and sPS. Evidence of this is the altered morphology of CMM blends. Here the DSC experiments also showed changed crystallization behavior. More graphic are the morphology light microscopy pictures. While, after melting and re-crystallization, the extruded samples show morphology characteristics for roughly separated polymer blends, in a CMM sample the iPP is finely distributed within the sPS.

ConclusionCMM using the Freezer/Mill causes changes to structure, thermal behaviour and morphology of homopolymers and polymer compounds. While morphological differences through melting and re-crystallization are partly reversible, structural changes remain. Aside of chain fission and the resulting macro radicals, cross-linking and reduced crystallization level has been observed. Polymer blends produced by CMM show a considerably improved dispersion of both polymers within each other. Exactly this changed degree of dispersion gives reason to hope for new materials with new and perhaps even predictable characteristics.

Application Note SP008: Structural Changes in Polymers :: APPLICATION: Polymer Blending

SP008


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Application Note SP015: RNA Extraction :: APPLICATION: Molecular Effects of Pollution

Materials and MethodsWe have devised the following multi-step approach in preparing our samples and using the SPEX Freezer/Mill.

• All SPEX microvials and other utensils used to extract the sample from the Freezer/Mill are autoclaved before use to avoid sample contamination.

• The SPEX Freezer/Mill is pre-chilled before the grinding of the samples using liquid nitrogen.

• The tissue samples are sliced into smaller pieces to improve the grinding.

• Sliced tissue samples are pre-chilled in liquid nitrogen in order to prevent RNA degradation.

• The pre-chilled samples are placed into a SPEX microvial and kept in liquid nitrogen until placement into the SPEX Freezer/Mill.

• The SPEX Freezer/Mill is topped off with liquid nitrogen and the vials containing samples are put into the SPEX Freezer/Mill.

• The following grinding process has shown to be effective in preventing heating the samples. The SPEX Freezer/Mill is turned on for 2 minutes and then off for 1 minute and the cycle is repeated for a total of 10 to 15 minutes of grinding.

• Once the samples have been ground, they are kept in the SPEX microvials in liquid nitrogen until further analysis.

• Samples are then ready for total RNA isolation by phenol and guanidium thiocyanate extraction.

ConclusionThe SPEX Freezer/Mill has been a substantial asset to our research by allowing the isolation of total RNA from our small tissue samples with improved quality and less degradation than previous techniques.

ReferencesGodard et al. (2004). Induction of Cetacean Cytochrome P4501A1 by -naphthoflavoneExposure of Skin Biopsy Slices. Toxicological Sciences. 80, 268-275.

Dr. Celine Godard-Codding and Carolyne LaCerte

SP015

Molecular Effects of Pollution in Cetacean Samples Using the Cytochrome P450, CYP 1A1, 1A2, and 1B1 as BiomarkersWith kind permission of Dr. Celine Godard-Codding, Ocean Alliance & The Whale Conservation Institute, Lincoln, MA

Ocean Alliance is dedicated to the conservation of whales and the ocean environment through research and education. Ocean Alliance recently completed a five-year research voyage around the world known as the Voyage of the Odyssey (2000 to 2005). The Voyage of the Odyssey was launched to address the need for a globally integrated dataset allowing a consistent analysis of exposure to, and potential effects of, persistent organochlorines and other pollutants in marine life.

Many marine mammals harbor large fatty reserves in their body where high levels of organochlorines and other lipophilic contaminants can accumulate.Marine mammals are subject to bioaccumulation and biomagnifications of those fat-soluble contaminants due to their relatively long life span and high trophic position within marine food chains. Therefore marine mammals can be considered environmentally relevant candidates for use as sentinel species when assessing marine pollution. The sperm whale (Physeter macrocephalus) was chosen as the indicator species for the Voyage of the Odyssey.A total of 960 sperm whale skin and blubber samples and over 100 samples from predatory fish were collected during the voyage.

Biomarker ProjectThe toxicology program of the Voyage of the Odyssey encompasses many projects. The biomarker project focuses on the use of the cytochrome P450 1 (CYP1) enzymes. CYP1 enzymes can metabolize and/or activate environmental pollutants, such as polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs). In many animal species CYP1 induction is used as a biomarker of exposure and molecular effects to the above contaminants. We recently validated the CYP1 biomarker in cetaceans (Godard et al., 2004).

The goal of our project is the investigation of the molecular effects of pollution in cetacean samples using the cytochrome P450 CYP 1A1, 1A2, and 1B1 as biomarkers. Our methods include isolating RNA sequencing, cloning and quantifying the expression of the 3 CYP genes in sperm whale skin biopsies. The sperm whale skin samples available for this particular biomarker project weigh only between 50-100ug. The amount of total RNA that can be extracted from such small samples is low; therefore we investigated the use of the SPEX Freezer/Mill in order to increase the quantity of total RNA that can be extracted from each sample and its quality.

Application Note SP015: RNA Extraction :: APPLICATION: Molecular Effects of Pollution

SP015


Sec tion 3 :


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Li2B4O7. This indicates that the end product of the reaction does not depend on the composition of the mixture but the conditions on the boundary layers.

ConclusionHigh-energy ball milling is especially suitable for particle size reduction and the study of subsequent chemical and physical changes. The characteristics of particle size reduction and subsequent growth was similar with all analyzed oxides. Some materials showed phase transformations while others showed chemical reactions, which did not occur with the microcrystalline starting material.

Application Note SP003: Nano Technology :: APPLICATION: Production of Nano Powders

SP003

Production of Nano-Crystalline Oxidized Ceramics with High-Energy Ball MillingS. Indris, D. Bork, P. Heitjans, J. Mater. Synth. Process 8, 245 (2000) With kind permission of Prof. P. Heitjans, Institute for Physical Chemistry and Electro-Chemistry, University of Hannover

Analyzing particle size effects in nanocrystalline materials requires a technique in which one can adjust the particle size. In this study various nano-crystalline materials were produced using a ball mill (8000M Mixer/Mill®, SPEX SamplePrep; equipped with alumina and zirconia vials). Ball milling is particularly suitable for this task as it is easy to use and allows the grinding of a relatively large amount of material as well as a large variety of different materials.

The analyzed media were: Li2O, LiNbO3, LiBO2, B2O3, TiO2 and Li2O:B2O3 mixtures. The average particle size was determined by the grinding time and subsequently analyzed by means of X-ray Diffraction (XRD) and Transmission Electron Microscopy (TEM). The lithium containing materials were selected because of their potential use as solid electrolytes. TiO2 is interesting with regard to its use as a photo catalyst. For hygroscopic materials the corundum grinding vial was filled in an argon atmosphere and put into an airtight stainless steel container.

Particle SizesThe different oxides displayed different grinding characteristics, but a minimum particle size of approx. 20nm was obtained after 8 to 10 hours of grinding. Particle size was determined by XRD analysis and TEM data. The nano-crystalline samples are metastable and heating led to particle growth, which was shown by DSC. This was taken into account during the sintering process when a solid, compact ceramic was to be produced from the samples. Prior studies by other groups showed the particular suitability of two-step sintering with a lower temperature during the second step.

TiO2, which was analyzed by two methods, showed partial phase transformation during the grinding process. When subjected to ball milling Rutile, comprising additional impurities, was obtained as pure Rutile (without impurities) in smaller particle sizes. Anatas, which showed the same impurities, exhibited an elevated proportion of these impurities after grinding. Therefore TiO2 has only limited applications in high-energy ball milling experiments.

Chemical ReactionsMixing of ceramic components and subsequent pressing produces a heterogeneous material with a multitude of different boundary layers. This network of different interfaces can be modified by varying the particle size. During analysis of a 50:50 mix of Li2O:B2O3 chemical changes due to this chemical-mechanical procedure were detected. After a short period of time only the lines of the original compounds are detected with XRD analysis, while new lines appear after 4 hours. The newly formed product was

Application Note SP003: Nano Technology :: APPLICATION: Production of Nano Powders

SP003


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Calculation % Crude Fat = Dry Extract Wt.(g)x100 (Note7) (asis) SampleWt.(g)

1 The material for extraction should be at or below equilibrium moisture. Wet materials should be dried by a method which does not result in fat oxidation.

2 Hexane is a very flammable solvent and all work should be performed in a well ventilated area with no open flames. Always insure that the hood is functioning properly before use.

3 High oil content samples should be ground in a cutting mill (e.g., Brinkman Model ZM-1); low oil content samples may be ground in a cutting mill or attrition mill.

4 Alternatively, the flask can be weighed and used for solvent evaporation as described later, if desired. Also, take care that no hexane is aspirated out of the flask on low oil samples.

5 Care must be taken when evaporating hexane. If the hexane is heated at too high a rate, the evaporation will occur with occasional violent boiling which will result in the loss of crude fat or oil.

6 Prolonged evaporation or drying of the extract at elevated temperatures may cause fat oxidation and high results.

7 It is customary to report the result as % oil when analyzing corn germ samples.

Analytical Methods of the Member Companies of the Corn Refiners Association, Inc.

Accepted 4-19-89

HT002

How To Subject: Crude Fat (Hexane Extractables)

HT002

HOW TO Guidelines : Feedstuffs Analysis – Crude Fat (Hexane Extractables)

PrincipleLipids are recovered by simultaneous milling and extraction with hexane in a sealed ball mill vial.

ScopeThe method is applicable to feedstuffs, corn germ and other components derived from the milling of corn (Note 1).

Special Apparatus1 Mill: Model 8000 Spex Mixer/Mill, equipped with a Cat. No. 8001 hardened steel vial

for grinding, and both 12 mm and 6 mm steel balls.

2 Filter Funnel: Gleman Cat. No. 4201 (47 mm) or Cat. No. 4230 (47 mm), or equivalent.

3 Filter Paper: Whatman Cat. No. 7195-004, a cellulose nitrate 5 micron filter which is hexane compatible.

4 Bath: Steam-heated water bath in a well ventilated hood.

5 Oven: Forced air or convection oven, operating at 100 °C.

Reagents1 n-Hexane: Reagent Grade or HPLC Grade (Note 2)

ProcedureGrind a 50-100 g sample to a uniform particle size (10 mesh) in a grinder or cutting mill (Note 3). Mix the sample thoroughly and weigh accurately about 4-5 g into the steel grinding vial. Add one 12mm and two 6 mm steel balls and 25 mL of n-hexane. Seal the vial and mill in the Mixer/Mill for 15 mins.

Place a filter paper in the funnel and filter the milled sample slurry into a clean and dry vacuum flask (Note 4). Quantitatively transfer the residual oil from the steel milling vial and cap with the aid of three 10 mL portions of hexane, and wash residual oil from the sample with another three 10 mL portions of hexane. Transfer the filtered hexane and oil miscella into a dried and tared 150 mL beaker. Rinse the vacuum flask residue into the beaker with small portions of hexane.

Evaporate the solvent on a steam-heated water bath under the hood (Note 5). Place the beaker with residue in the air oven and dry it 1 hour at 100 °C (Note 6). Remove the beaker and cool in a desiccator. Weigh the beaker and calculate the dry extract weight.

How To Subject: Crude Fat (Hexane Extractables)


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Measured results were also consistent for the ten scans of each bead. An example for one bead is shown in Appendix 1 and is indicative of results obtained from repetitive scans of the other nine beads. Complete data for this study are on file at SPEX SamplePrep, LLC, Metuchen, NJ, USA.

Table 1. Fly Ash Composition (%) using FFB-5005-02

Analyte Average Bead 1 Bead 2 Bead 3 Bead 4 Bead 5

SiO2 52.08 52.11 52.07 52.11 52.08 52.06

Al2O3 26.75 26.74 26.76 26.74 26.74 26.76

Fe2O3 6.52 6.52 6.52 6.52 6.52 6.52

K2O 4.14 4.14 4.13 4.14 4.14 4.15

CaO 3.46 3.46 3.45 3.46 3.46 3.46

MgO 2.04 2.04 2.04 2.04 2.04 2.05

P2O5 1.42 1.42 1.43 1.42 1.43 1.43

TiO2 1.26 1.26 1.25 1.26 1.26 1.26

Na2O 1.23 1.22 1.25 1.22 1.24 1.23

SO3 0.16 0.16 0.16 0.16 0.16 0.16

Table 2. Fly Ash Composition (%) using FFB-6705-02

Analyte Average Bead 1 Bead 2 Bead 3 Bead 4 Bead 5

SiO2 52.07 52.05 52.12 52.09 52.09 52.02

Al2O3 26.75 26.75 26.79 26.75 26.69 26.76

Fe2O3 6.49 6.50 6.47 6.50 6.50 6.50

K2O 4.15 4.15 4.15 4.15 4.16 4.16

CaO 3.47 3.48 3.45 3.48 3.48 3.47

MgO 2.05 2.06 2.02 2.06 2.06 2.06

P2O5 1.42 1.41 1.41 1.41 1.42 1.42

TiO2 1.26 1.26 1.25 1.26 1.26 1.26

Na2O 1.24 1.24 1.23 1.24 1.24 1.25

SO3 0.16 0.16 0.16 0.16 0.16 0.16

APPLICATION NOTE SP026: Preparation and XRF Analysis of Fly Ash Fused BeadsAPPLICATION: Cement, Slag & Fly Ash

SP026

Preparation and XRF Analysis of Fly Ash Fused Beads

ABSTRACTFly ash is a waste product from the combustion of coal and is comprised of the mineral particles that rise with the flue gases. Recovered fly ash is used as a component in certain cement mixes and improves the durability and strength of concrete.

In this study, a sample of fly ash was pulverized and blended, and then fused into two sets of glass beads using a Katanax K1 electric fusion fluxer. Two different lithium borate fusion fluxes from SPEX CertiPrep were used to prepare five beads with each flux. The beads were then analyzed by energy dispersive X-ray fluorescence spectroscopy (ED-XRF) and the results were compared both within a set and between the two sets to evaulate consistency of beads produced using the K1 and effect of flux composition on the XRF measurements.

Fusion MethodThe fly ash was roasted in a porcelain crucible for 2 hr. at 750° C to eliminate residual carbon (~0.2%). 6.40 g of flux and 0.80 g of fly ash (a flux/sample ratio of 8:1) were weighed into a Pt/Au crucible, gently mixed by hand with a plastic laboratory spatula, and fused into a 32 mm glass bead using the K1. The fusion program used was a modified version of the K1’s fixed OXIDE program, with the main fusion stage (Step 4) set for 15 minutes at 1035° C.

Five beads were prepared in this manner for each of two fluxes: SPEX CertiPrep FFB-5005-02 (1:1 lithium tetraborate:lithium metaborate with 0.5% lithium bromide as a non-wetting agent) and FFB-6705-02 (2:1 lithium tetraborate:lithium metaborate with 0.5% lithium bromide).

AnalysisThe two sets of glass beads were analyzed by ED-XRF. Each bead was measured ten times, with the results averaged for each bead. The results for the five beads within a set were then averaged again to obtain overall results for the set. The results are shown in Tables 1 & 2 as percentage of composition for each analyte, with the major components being silica and alumina. Oxides of iron, potassium, calcium, magnesium, phosphorus, titanium, sodium, and sulfur were present in lesser amounts and comprised the remainder of the fly ash.

XRF results show excellent consistency within a set, indicating that the fly ash sample was homogenous and that the K1 prepares fused beads with a high level of reproducibility. In addition, the overall averages from Table 1 and Table 2 are nearly identical. Thus, it is apparent that the choice of flux composition had no effect on the XRF results. Since both fluxes produced clear glass beads with the fly ash sample, either is appropriate to use with this fly ash.


SP026


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Appendix

ConclusionThe Katanax K1 electric fusion fluxer is easy to operate and produces high quality, reproducible glass beads. The two fluxes used in this study gave XRF results that indicate that flux choice did not influence the analysis. In addition, the high degree of bead to bead consistency within a set indicates that the elemental composition of the flux is uniform.

Table 3. Fly Ash results for 10 successive XRF scans of Bead 4 prepared using FFB-5005-02.

Analyte SiO2 Al2O3 Fe2O3 K2O CaO

Scan 1 52.09 26.81 6.52 4.14 3.47Scan 2 52.05 26.81 6.51 4.13 3.46Scan 3 52.14 26.71 6.52 4.15 3.45Scan 4 52.04 26.71 6.51 4.15 3.47Scan 5 52.07 26.80 6.51 4.14 3.48Scan 6 52.11 26.76 6.53 4.12 3.46Scan 7 52.04 26.71 6.53 4.15 3.47Scan 8 52.05 26.75 6.51 4.16 3.42Scan 9 52.14 26.67 6.52 4.15 3.48Scan 10 52.07 26.67 6.52 4.15 3.46Average 52.08 26.74 6.52 4.14 3.46

Analyte MgO P2O5 TiO2 Na2O SO3

Scan 1 2.04 1.42 1.26 1.18 0.16Scan 2 2.02 1.42 1.26 1.24 0.16Scan 3 2.04 1.42 1.27 1.21 0.16Scan 4 2.05 1.45 1.26 1.29 0.16Scan 5 2.02 1.42 1.26 1.23 0.16Scan 6 2.03 1.42 1.26 1.20 0.16Scan 7 2.07 1.46 1.26 1.24 0.16Scan 8 2.05 1.46 1.26 1.27 0.16Scan 9 2.07 1.42 1.27 1.21 0.16Scan 10 2.03 1.42 1.25 1.31 0.16Average 2.04 1.43 1.26 1.24 0.16


SP026

Order Online 24/7: www.spexsampleprep.com204 Order Online 24/7: www.spexsampleprep.com 205

Appendix

XRF Sample Preparation: Selected References

Reference Books Bennett, H. and Oliver, G., XRF Analysis of Ceramics, Minerals and Allied Materials, JohnWiley & Sons, New York, 1992. The most thorough, practical work about borate fusions, with methods for specific types of samples.

Bertin, E., Principles and Practice of X-Ray Spectrometric Analysis, 2nd ed., Plenum Press, New York, 1975. Much useful information throughout on sample preparation methods and equipment.

Bock, R., Decomposition Methods in Analytical Chemistry, International Textbook Co., Glasgow, 1979. Authoritative text on all decomposition techniques with many fusion methods given.

Buhrke, V., Jenkins, R. and Smith, D., A Practical Guide for the Preparation of Specimens for X-ray Fluorescence and X-ray Diffraction Analysis, JohnWiley & Sons, Inc., New York, 1998. Comprehensive reference for XRF and XRD sample preparation.

Claisse, F. and Blanchette, J. S., Physics and Chemistry of Borate fusion: Theory and Application, Fernand Claisse, Inc., Quebec, P.Q., Canada, 2004. A compact, essential guide to the theory and practice of borate fusions.

Jenkins, R., Gould, R.W., and Gedcke, D., Quantitative X-Ray Spectrometry, Marcel Dekker, Inc., New York, 1981. Chapter 7, Specimen Preparation, pgs. 363–394.

Sulcek, Z. and Povondra, P., Methods of Decomposition in Inorganic Analysis, CRC Press, Inc., Boca Raton, 1989. Many specific fusion methods. Lachance, G., and Claisse, F., Quantitative X-Ray Fluorescence Analysis: Theory and Application, JohnWiley & Sons., New York, 1995. Section 9.3, Specimen Preparation, pgs. 245–251.

Van Grieken, R., and Markowicz, A., Editors, Handbook of X-Ray Spectrometry: Methods and Techniques, Marcel Dekker, Inc., New York, 1993. Chapter 13, Sample Preparation for XRF.

Works of a Periodical Nature Advances in X-ray Analysis, Vols. 1 (1957) to the present, available on CDROM and through the website of the International Centre for Diffraction Data®: www.icdd.com. The annual proceedings of the Denver X-ray Conference, with papers on every aspect of X-ray analysis, including sample prepartion.

X-Ray Spectrometry, a bimonthly journal published by JohnWiley & Sons Ltd., Baffins Lane, Chichester,West Sussex, England. Occasional articles on sample preparation.

Most of the above publications have detailed bibliographies for further reading.










2010 Geno/Grinder

8530 Shatterbox

3635 X-Press




Appendix


Appendix

Questionnaire for test grinding of Samples

Up to 3 samples will be ground at no charge, and a report and recommendations will be returned within three weeks.

SPEX SamplePrep Mill for Test: ______________________________________________

SPEX SamplePrep Grinding or Mixing container: ___________________________________

Sample Description and Composition: _________________________________________

If sample is incompatible with any grinding container or considered hazardous in any way please include MSDS

Purpose for Grinding: ___________________________________________________

Emission Spectroscopy: _____________ X-Ray Spectroscopy: ________________________

Extraction: ________________________ Other: _____________________________

Desired Particle Size after Grinding, if known: _____________________________________

If contamination with certain metals or plastics is objectionable, please list them along

with the approximate ppm level(s) of concern: ____________________________________

Notes or Comments: ____________________________________________________

For Return of Sample and Report: ____________________________________________

Name and Title: _______________________________________________________

Company Name: ______________________________________________________

Mailing Address: ______________________________________________________

Phone: ____________________________________________________________

Date: _____________________________________________________________

Mail to SPEX SamplePrep, 15 Liberty Street, Metuchen NJ 08840

Please contact SPEX SamplePrep before sending in your test sample: 1-732-623-0465: [email protected]

Samples are required to be sent on customers account, excess sample that is considered to be hazardous will be returned.

Trademark References

Registered Trademarks of SPEX SamplePrep, LLC™:• AutoExtractor • Geno/Grinder • Spec-Cap• Bench-Press • Mixer/Mill • UltraBind• ElectroFluxer • PrepAid • Ultralene• Freezer/Mill • Shatterbox • X-Press

Registered Trademarks of SPEX CertiPrep, Inc.®:• Assurance • SPEXertificate• Certi-Strips • SPEX CertiPrep Ltd• Claritas PPT • Triple Checked for Quality• Enviro-PT • VAL-U-CAL

Registered Trademarks of International Centre for Diffraction Data®:• ICDD

Registered Trademarks of Applied Biosystems:• TaqMan

Registered Trademarks of E.I. DuPont de Nemours & Co.:• Delrin • Kapton• Teflon • Vertrel XF

Registered Trademarks of DuPont Teijin Films:• Mylar

Registered Trademark of Katanax Inc:• Katanax

Registered Trademarks of Qiagen:• QIAshredder• DNeasy

Registered Trademarks of Roche Molecular Systems and Hoffman LaRoche:• PCR

Registered Trademarks of Promega Corporation:

• Wizard


Bench Press, 56

Binders, 62-63, 75-76, 99, 117-118

Borate Fusion Flux, 72

Bubble -Free Insert, 76, 131-132

Cap -Mats, 22-23

Cellulose Binder, 62-63, 75-76, 117-118

Centrifuge Tube Holder, 27

Ceramic Grinding Media, 21

Certified Reference Materials, 81-84

Chemical Attack on Window Films, 133-138

Clamp Assembly for Geno/Grinder, 19

Cleaning Grinding Containers, 99-100

Composition of Grinding Containers, 90

Contamination, 98

Cr- Free Vial, 34-35

Critical (Infinite) Thickness, 111-112

Crucibles Graphite, 69-72, 128-129 Platinum, 66 Zirconium, 67

Crucible Tongs, 67

Cryo- Block, 28-29

Cryogenic Gloves, 38

Cryogenic Grinding, 86, 92-96

Cryo Station, 28, 146-147

Cryogenic Transfer Hose, 39

Demo Program, 16, 140, 204

Dewar, 38-39

Die Sleeve and Plunger, 60-61, 119

Dies, Evacuable, for Pelletizing, 59, 114-116

Adapters, for Freezer/Mill, 36 for Mixer/Mill, 49

Agate, Balls, 48 Composition, 90 Usage, 97 Vial for Mixer/Mill, 44, 46

Air- Actuated Press, 56

Alumina Ceramic, Balls, 48 Composition, 90 Container for Shatterbox, 52 Grinding Application, 97 Usage, 97 Vial for Mixer/Mill, 45

Application Notes, Geno/Grinder, 142-183 Freezer/Mill, 184-195 Mixer/Mill, 196-199 Katanax, 200-202

AutoExtractor, 35

Automated Press, 55

Auto Fill System, 38

Ball Dispenser, 19

Balls, Agate, 48-49 Alumina Ceramic, 48 Methacrylate, 48 Silicon Nitride, 48 Stainless Steel, 19-20, 48, Tool Steel, 48 Tungsten Carbide, 48 Zirconia, 48

Beads, Silica, 20-21, 25 Zirconium, 21, 25-26

S ubjec t IndexS ubjec t Index


Kapton XRF Window Film, 78, 80, 133

KBr Dies, 59, 123

KBr Mulls, 100

Laboratory Presses, 55-58

Liquid Sample Cells for XRF, 76-77, 130-131

Liquids, XRF Analysis of, 130-131

Lithium Borate Fluxes, 72, 126-127

Magnet, 19

Manual Presses, 56-58

Mechanical Alloying, 100-101, 104

Methacrylate, Balls, 48 Usage, 97 Vials for Mixer/Mill, 43-45, 47

Micro X -Cells, 77, 13

Microvial, 33, 93 Microvial Extraction Tool, 35

Mills, Freezer/Mill, 30-32 Mixer/Mill, 40-41 Shatterbox, 50-51

Minimum Orders, 5-6

Mixer/Mills, 40-42, 102-104

Molds, 67-68

Motorized Presses, 55, 110

Mulls, 100

Mylar XRF Window Film, 78, 80, 120, 133, 136

Non Wetting Agents, 73, 127-128

Ordering Information, 3-4

Paraffin Binder, 63, 76, 117-118

Particle Size Effects in XRF, 111-112

Dispenser, 19 Disposable XRF Liquid Cells, 76-78, 130-132

DNA Extraction, 92, 153-155

Dual Mixer/Mill, 40, 103-104

Edge Flaring Tool, 62, 75

Edge Flaring Tool, 62, 75, 117

Enclosed Shatterbox, 50, 107-108

Extractor & Vial Opener, 35-36

Films, XRF Window, 78-80, 132-138

Flux, 72, 126-127

Fluxers, 64-65

Freezer/Mill, 30-32

Fusion, 64-73, 124-129

Fusion Fluxes, 72, 126-127

Fusion Machines, 64-65, 124

Geno/Grinder, 18, 86

Gloves, Cryogenic, 38

Graphite Crucibles, 69-72, 128-129

Grinding Aids, 49, 54, 98-99

Grinding Containers, 52-53

Grinding Key Points, 97-101

Hardened Steel, Composition, 90 Containers for Shatterbox, 52-53 Usage, 97 Vials for Mixer/Mill, 44-46

Hardness of Grinding Container Materials, 90

How To Order, 3-4

Hydraulic Presses, 55-58

Infinite (Critical) Thickness, 111-112

Infrared Mulls, 100



Return Shipment, 6

RoHS/WEEE, Analysis, 83, 93-95 Vials, 34-35

Sample Cells, Liquid, for XRF, 74, 76-78, 110, 121, 130-133

Shatterbox, 50-51, 107-109

Shatterbox, Grinding Containers, 52-53, 109

Shipping, 5

Silica Grinding, Beads, 20-21 Pre-Filled Grinding Vials, 25-26 Resin, 21

Silicon Nitride, Grinding Media, 49 Usage, 97 Vials for Mixer/Mill, 46

Sleeve- and- Plunger, 60-61, 110-111, 119

Snap -On Rings for X- Cells, 76-77

Solid Reference Materials, 84

Spec Caps, 61-62, 74-75, 99, 110-111, 113, 116-117, 130

SPEX SamplePrep Carver Presses, 58-59, 110, 114

Stainless Steel, Composition, 90-91 Grinding Media, 20, 27, 48 Pellets, 87 Usage, 97, 100, 102-103 Vials for Freezer/Mill, 33-34, Vials for Mixer/Mill, 43, 45-47

Thin Films for XRF, 78-80, 132-138

Titer Plates, 22-23

Transmission, of XRF Window Films, 134

Transporter, 54

Pellet Caps, for XRF (Spec Caps), 74-75, 116-117, 130

Pellet Dies, 59-61, 115-116, 123

Pelletizing, 55-63, 110-123

Pellets for Pellet Dies, 60

Plastic, Grinding Media, 48 Films for XRF, 78-79, 132-138 Vial Comparison, 47 Vials for Mixer/Mill, 43-44, 97

Platinumware, 67-68

Polycarbonate, Center Section, 33-34, 185-186 Vials for Freezer/Mill, 33-35 Vials for Geno/Grinder, 24-25, 27 Vials for Mixer/Mill, 43

Polyethylene Vials for Mixer/Mill, 46

Polypropylene, XRF Window Film, 79-80, 132-134, 137

Polystyrene, Usage, 97 Vials, Comparison, 90-91 Geno/Grinder, 23-27 Freezer/Mill, 33-35 Mixer/Mill, 43-46, 97-98

Powder Samples, Preparation for XRF, 110-111

Pre -Flared Spec Caps, 61, 74, 117

PrepAid, 49, 54, 62-63, 75-76, 98-99, 117

Presses, 55-58, 110

Pressing, 55-63, 110-123

Propylene Glycol Grinding Aid, 49, 54

Rack, Shatterbox, 54 Freezer/Mill, 35, 37



Zirconia, Balls, 48 Composition, 90 Container Usage, 107 Container, 52 Vial, 45, 47, 97, 196

Zirconium, Beads, 21 Composition, 90 Crucibles, 67 Heater Lid, 68 Pre-filled Vials, 25-26

Tungsten Carbide, Balls, 48 Composition, 90 Containers, 52-53 Pellets, 60 Usage, 97-98, 107 Vials for Mixer/Mill, 44-45

UltraBind, 62, 76, 110, 113, 117-118,

Ultralene XRF Window Film, 79, 80, 132-133,

Vertrel XF, 49, 54, 99

Vials Adapter, For Freezer/Mill, 36-37, 187 For Mixer/Mill, 49, 102

Vials, Agate, 44, 46-47, 90, 97, 100, 102-103 Alumina Ceramic, 45, 47, 90, 97, 103, 196, Cr -Free, 34-35, 90, 93-94 Hardened Steel, 44, 46-47, 97, 99-100, 102-103, 198-199 Plastic, 43, 44, 46-47, 90 Racks, 27, 35, 37, Silicon Nitride, 46-47, 90, 97 Stainless Steel, 33-34, 43, 45-47, 90, 97, 100, 102-103, 196 Tungsten Carbide, 44-45, 47, 90, 97, 102-103, 107 Zirconia, 45, 47, 90, 97, 196

Warranty, 6

Window Film, for XRF Cells, 78-80, 132-138

X- Cells, 76-77, 110, 121, 130-133

X -Press, 55, 99-100, 114

X- Ray Transmission, Thin Films, 78-80, 132-138

XRF, Accessories, 74-80, 130-138 Liquid Sample Cells, 76-77, 130-131 Pellet Caps (Spec Caps), 61-62, 74-75, 99, 110-111, 113, 116-117, 130 Window Films, 78-80, 132-138

Yeast Extractions, 86, 146-147, 151-152



2211 Cap-Mat for 2210 Titer Plate ............................................................................... 22 2211-10 Cap-Mat for 2210 Titer Plate, Case of 10 ............................................................. 222220 48-Well Titer Plate .............................................................................................. 23 2220-100 48-Well Titer Plate, Case of 100 .......................................................................... 222221 Cap-Mat for 2220 Titer Plate ............................................................................... 23 2221-10 Cap-Mat for 2220 Titer Plate, Case of 10 ............................................................. 232230 24-Well Titer Plate .............................................................................................. 23 2230-100 24-Well Titer Plate, Case of 100 .......................................................................... 232231 Cap-Mat for 2230 Titer Plate ............................................................................... 23 2231-10 Cap-Mat for 2230 Titer Plate, Case of 10 ............................................................. 232231-10 Cap-Mat for 2230 Titer Plate, Case of 10 ............................................................. 232240-PC Pre-Cleaned Polycarbonate Vial Set, 5 mL, Case of 10 ........................................ 232240-PEF Pre-Cleaned Frosted Polyethylene Vial Set, 5 mL, Case of 10 .............................. 23 2241-PC Polycarbonate Vial, 5 mL .................................................................................... 23 2241-PEF-200 Frosted Polyethylene Vial, 5 mL .......................................................................... 242250 Pre-Cleaned Polycarbonate Vial Set, 15 mL ........................................................ 24 2251PC Polycarbonate Vial, 15 mL, Short ........................................................................ 24 2252-PC-30 Polycarbonate Vial, 15 mL, Tall ........................................................................... 24 2253C-48 Cap for 2253-PC-48 ............................................................................................ 24 2253-PC-48 Polycarbonate Vial, 50 mL .................................................................................. 24 2254-PE-48 LDPE Vial, 50 mL ................................................................................................. 252255 Cryogenic Grinding Vial Pack .............................................................................. 25 2255C20 Polycarbonate Center Cylinder, pack of 20 .......................................................... 25 2255C4 Polycarbonate Center Cylinder, pack of 4 ............................................................ 252260 Cryo-Block for 2255 Vial, set of 2 ........................................................................ 28 2300 Holder for 2 mL microfuge Tubes ........................................................................ 272301-100MB 2 mL Tube with 100 μm Molecular Biology Grade Silica Beads ........................... 25 2301-400MB 2 mL Tube with 400 μm Molecular Biology Grade Silica Beads ........................... 25 2301-800MB 2 mL Tube with 800 μm Molecular Biology Grade Silica Beads ........................... 25 2302-1000AW 2 mL Tube with 1.0 mm Acid Washed Zirconium Beads ...................................... 26 2302-100AW1 2 mL Tube with 100 μm Acid Washed Zirconium Beads ...................................... 25 2302-100AW2 2 mL Tube with 100 μm Acid Washed Zirconium Beads ...................................... 25 2302-100LB 2 mL Tube with 100 μm Low Binding Zirconium Beads ...................................... 25 2302-1400AW 2 mL Tube with 1.4 mm Acid Washed Zirconium Beads ...................................... 26 2302-1700AW 2 mL Tube with 1.7 mm Acid Washed Zirconium Beads ...................................... 26 2302-200AW 2 mL Tube with 200 μm Acid Washed Zirconium Beads ...................................... 26

Product Number Product Name Page Number2010 Geno-Grinder ..................................................................................................... 18 2100 Grinding Ball Dispenser ...................................................................................... 19 2110M 24-Pin Magnet ................................................................................................... 19 2110M-12 12 Pack of 12 Magnetic Tips................................................................................ 20 2110M-96 96 Pack of 96 Magnetic Tips................................................................................ 20 2110S Separator for 24-Pin Magnet .............................................................................. 192150 Grinding Balls, 5/32 in. (4 mm ........................................................................... 20 2155 Grinding Balls, 3/8 in. (9.5 mm .......................................................................... 20 2156 Grinding Balls, 7/16 in. (11 mm ......................................................................... 20 2160 Silica Grinding Beads, 800-1000 μm .................................................................. 20 2161 Silica Grinding Beads, Molecular Biology Grade, 800-1000 μm .......................... 20 2162 Low Binding Silica Beads, 800 μm ...................................................................... 20 2165 Silica Grinding Beads, 400-600 μm .................................................................... 20 2166 Silica Grinding Beads, Molecular Biology Grade, 400-600 μm ............................ 20 2167 Low Binding Silica Beads, 400 μm ...................................................................... 21 2168 Low Binding Silica Beads, 100 μm ...................................................................... 21 2170 Silica Grinding Resin, 100-400 μm ..................................................................... 21 2171 Silica Grinding Resin, Molecular Biology Grade, 100-400 μm ............................. 21 2180 Zirconium Grinding Beads, Molecular Biology Grade, 200-400 μm .................... 21 2181 Low Binding Zirconium Beads, 100 μm .............................................................. 21 2182 Low Binding Zirconium Beads, 200 μm .............................................................. 21 2183 Ceramic Grinding Cylinder, 3/8 in. x 7/8 in ......................................................... 21 2184 Ceramic Grinding Cylinder, 5/16 in. x 5/8 in ....................................................... 21 2185 Ceramic Grinding Cylinder, 5/32 in. x 5/16 in ..................................................... 21 2190 Vial Rack ............................................................................................................. 27 2196-16-PE Holder for 50 mL Centrifuge Tubes ..................................................................... 272197 Holder for 15 mL Centrifuge Tubes ..................................................................... 27 2198-24-PE Holder for 6133PC Vial ........................................................................................ 272199 Large Capacity Clamp Assembly ......................................................................... 19 2200 96-Well Titer Plate, Square Wells ........................................................................ 22 2200-100 96-Well Titer Plate, Square Wells, Case of 100 .................................................... 222201 Cap-Mat for 2200 Titer Plate ............................................................................... 22 2201-10 Cap-Mats for 2200 Titer Plates, Case of 10 .......................................................... 222210 96-Well Titer Plate, Round Wells......................................................................... 22 2210-100 96-Well Titer Plate, Round Wells, Case of 100 ..................................................... 22

Produc t IndexProduc t Index


3520 Polypropylene Window Film (Roll ...................................................................... 79 3521 Polypropylene Pre-Cut Circles 0.2 mil (5 μm) thick ............................................. 79 3524 Micro-porous Teflon® Film (Roll .......................................................................... 79 3525 Ultralene® Window Film (Roll ............................................................................ 79 3526 Ultralene® Pre-Cut Circles ................................................................................... 79 3527 X-Cell, 40 mm Closed X-Ray Cell ......................................................................... 76 3527I Bubble-Free Cell Insert ....................................................................................... 763529 X-Cell, 31 mm Closed X-Ray Cell ......................................................................... 76 3561 X-Cell, 31 mm Universal Closed X-Ray Cell with Collar........................................ 77 3565 X-Cell, 43 mm Closed X-Ray Cell For Horiba Sulfur Analyzers ............................. 77 3571 X-Cell, 31 mm Double Open End X-Ray Cell with Collar ...................................... 77 3577 X-Cell, 31 mm Micro X-ray Cell with Collar.......................................................... 77 3610 Die Set – 10 mm ................................................................................................ 59 3613 Die Set – 13 mm ................................................................................................ 593613ST Steel Pellets – 13 mm ........................................................................................ 603614 Die Set – 40 mm ................................................................................................ 59 3614C Tungsten Carbide Pellets – 40 mm ..................................................................... 60 3614ST Steel Pellets – 40 mm ........................................................................................ 603614W Die Sleeve-and-Plunger Set – 40 mm ................................................................ 613615 Spec-Cap – 33 mm ............................................................................................. 613615M Spec-Cap – 33 mm ............................................................................................. 61 3615 33 mm Spec-Cap ................................................................................................ 75 3616 Die Set – 35 mm ................................................................................................ 593616C Tungsten Carbide Pellets – 35 mm ..................................................................... 60 3616ST Steel Pellets – 35 mm ........................................................................................ 60 3616W Die Sleeve-and-Plunger Set – 35 mm ................................................................ 61 3617 Spec-Cap – 38 mm ............................................................................................. 62 3617 38 mm Spec-Cap ................................................................................................ 75 3617M Spec-Cap – 38 mm ............................................................................................. 62 3617M 38 mm Spec-Cap ................................................................................................ 753618 Edge-Flaring Tool ................................................................................................ 62 3618 Edge-Flaring Tool ................................................................................................ 75 3619 Spec-Cap® - 30 mm ............................................................................................ 61 3619 30 mm Spec-Cap® .............................................................................................. 74 3619A Pre-Flared Spec-Cap – 31 mm ........................................................................... 61 3619A 31 mm Pre-Flared Spec-Cap ............................................................................... 74

2302-3000AW 2 mL Tube with 3 mm Acid Washed Zirconium Beads ......................................... 26 2303-MM1 2 mL Tube with Mixed Matrix 1 .......................................................................... 26 2303-MM2 2 mL Tube with Mixed Matrix 2 .......................................................................... 26 2303-MM3 2 mL Tube with Mixed Matrix 3 .......................................................................... 26 2304-100AW 2 mL Tube with 100 μm Acid Washed Silica Beads ............................................. 26 2304-400AW 2 mL Tube with 400 μm Acid Washed Silica Beads ............................................. 26 2304-800AW 2 mL Tube with 800 μm Acid Washed Silica Beads ............................................. 26 2305-2800SS 2 mL Tube with 2.8 mm Stainless Steel Balls ...................................................... 272600 Cryo-Station ....................................................................................................... 28 2650 Cryo-Adapter for Titer Plates .............................................................................. 28 2660 Cryo-Block for 15 mL Polycarbonate Vials........................................................... 29 2661 Cryo-Block for 15 mL Centrifuge Tubes ............................................................... 29 2662 Cryo-Block for 4 mL Polycarbonate Vials............................................................. 29 2663 Cryo-Block for 4 mL Polyethylene Vials............................................................... 29 2664 Cryo-Block for 50 mL Centrifuge Tubes ............................................................... 29 2665 Cryo-Block for Micro-centrifuge or PCR Tubes .................................................... 29 2666 Cryo-Block for 48 Micro-centrifuge or PCR Tubes ............................................... 29 3111 Polystyrene Vial .................................................................................................. 43 3112 Methacrylate Balls .............................................................................................. 48 3114 Stainless Steel Vial Set ........................................................................................ 43 3114SB Stainless Steel Ball Set ........................................................................................ 483116 Polystyrene Vial .................................................................................................. 43 3116PC 5 mL Polycarbonate Vial ..................................................................................... 27 3116PC Polycarbonate Vial .............................................................................................. 433117 Hardened Tool Steel Vial Set ............................................................................... 44 3117B Hardened Tool Steel Ball Set ............................................................................... 483118 Agate Vial Set ..................................................................................................... 44 3118A Agate Ball ........................................................................................................... 483119 Methacrylate Balls .............................................................................................. 48 3127 Hardened Tool Steel Vial Set ............................................................................... 44 3511 Kapton® Window Film (Roll ................................................................................ 78 3512 Kapton Pre-Cut Circles ........................................................................................ 78 3515 Mylar® Pre-Cut Circles (Large ............................................................................. 78 3516 Mylar Window Film (Roll .................................................................................... 78 3517 Mylar Window Film (Roll .................................................................................... 78 3518 Mylar Pre-Cut Circles .......................................................................................... 78



6752 Small Steel Center Cylinder ................................................................................. 33 6753 Microvial Set ....................................................................................................... 33 6753C Microvial Sample Extraction Tool ........................................................................ 356754 Extractor/Vial Opener for Small Vials .................................................................. 36 6755 Vial Rack for Small Vials ...................................................................................... 35 6761 Small Poly-Vial Set ............................................................................................. 33 6770 SPEX SamplePrep 6770 Freezer/Mill® ................................................................. 32 6771 Small Cr-Free Vial Set ......................................................................................... 34 6781S Small Stainless Steel Grinding Vial Set ............................................................... 346801 Large Grinding Vial Set ....................................................................................... 346801C20 Large Polycarbonate Center Cylinder .................................................................. 34 6801C4 Large Polycarbonate Center Cylinder .................................................................. 346802 Large Steel Center Cylinder ................................................................................. 34 6804 Extractor/Vial Opener for Large and Mid-Sized Vials .......................................... 36 6805 Vial Rack for Large Vials ...................................................................................... 35 6806 Multi-Vial Adapter .............................................................................................. 36 6814 AutoExtractor™ for Freezer/Mill® Vial Sets.......................................................... 35 6820 Auto-Fill System for the 6870 Freezer/Mill ......................................................... 38 6870 SPEX SamplePrep 6870 Freezer/Mill® ................................................................. 31 6870L Large Vial Accessory Package.............................................................................. 37 6870M Mid-Size Vial Accessory Package ........................................................................ 37 6870S Small Vial Accessory Package ............................................................................. 376871 Large Cr-Free Vial Set .......................................................................................... 34 6881 Mid-Size Vial Set ................................................................................................. 34 6881C20 Mid-Size Polycarbonate Center Cylinder ............................................................. 35 6881C4 Mid-Size Polycarbonate Center Cylinder ............................................................. 346883 Mid-Size Cr-Free Vial .......................................................................................... 35 6884 Mid-Size Vial Adapter for 6804 Extractor/Vial Opener ........................................ 36 6885 Mid-Size Poly-Vial .............................................................................................. 35 6886 Mid-Size Vial Adapter for 6800-Series Freezer/Mills ........................................... 36 6900 Cryogenic Gloves ................................................................................................ 38 6905 Portable Cryogenic Dewar, 25 L Capacity ........................................................... 38 6906 Short Cryogenic Transfer Hose ............................................................................ 39 6907 Long Cryogenic Transfer Hose ............................................................................. 39 6910 Portable Pressurized Cryogenic Cylinder, 50 L Capacity ...................................... 39 6970D SPEX SamplePrep 6970EFM Enclosed Freezer/Mill®– Dual Grinding Coils.......... 30

3619AM Pre-Flared Spec-Cap – 31 mm ........................................................................... 61 3619AM 31 mm Pre-Flared Spec-Cap ............................................................................... 74 3619M Spec-Cap – 30 mm ............................................................................................. 61 3619M 30 mm Spec-Cap ................................................................................................ 743621 3621 SPEX SamplePrep-Carver Model C ............................................................. 57 3622 3622 SPEX SamplePrep-Carver Model M ............................................................ 58 3623 Die Set – 31 mm ................................................................................................ 59 3623C Tungsten Carbide Pellets - 31 mm ...................................................................... 60 3623ST Steel Pellets – 31 mm ........................................................................................ 60 3623W Die Sleeve-and-Plunger Set – 31 mm ................................................................ 603625 Edge-Flaring Tool ................................................................................................ 62 3625 Edge-Flaring Tool ................................................................................................ 75 3626 3626 SPEX SamplePrep-Carver 12-ton Press ...................................................... 58 3628 SPEX SamplePrep 3628 Air-Actuated Bench-Press® ........................................... 56 3635 SPEX SamplePrep 3635 Automated X-Press® ..................................................... 55 3640 PrepAid Propylene Glycol ................................................................................... 49 3640 PrepAid Propylene Glycol ................................................................................... 54 3642-150 PrepAid Cellulose Binder ................................................................................... 62 3642-450 PrepAid Cellulose Binder ................................................................................... 62 3644-150 PrepAid UltraBind® ............................................................................................. 62 3644-450 PrepAid UltraBind® ............................................................................................. 623646-150 PrepAid Paraffin Binder – (CnH2n+2 ................................................................. 63 3646-450 PrepAid Paraffin Binder – (CnH2n+2 ................................................................. 633650 PrepAid Vertrel® XF ............................................................................................. 49 3650 PrepAid Vertrel® XF ............................................................................................. 54 5004 Tungsten Carbide-Lined Vial Set ......................................................................... 44 5004A Tungsten Carbide Ball Set ................................................................................... 48 5004C Methacrylate Center Cylinder ............................................................................. 445100 SPEX SamplePrep 5100 Mixer/Mill® ................................................................... 42 6133 Polystyrene Vial .................................................................................................. 436133PC Polycarbonate Vial .............................................................................................. 43 6134 Polystyrene Vial .................................................................................................. 46 6135 Polyethylene Vial ................................................................................................ 46 6751 Small Grinding Vial Set ....................................................................................... 33 6751C20 Small Polycarbonate Center Cylinder .................................................................. 33 6751C4 Small Polycarbonate Center Cylinder .................................................................. 33



8011 Multiple-Sample Adapter................................................................................... 49 8012 Vial Clamp Adapter for the 8000D Mixer/Mill ..................................................... 49 8014 Agate Vial Set ..................................................................................................... 46 8014A Agate Ball ........................................................................................................... 498015 Clamp for 8003 Vial Set ...................................................................................... 45 8500 SPEX SamplePrep 8500 Shatterbox® .................................................................. 51 8501 Hardened Steel Grinding Container .................................................................... 52 8504 Tungsten Carbide Grinding Container ................................................................. 52 8505 Alumina Ceramic Grinding Container ................................................................. 52 8506 Zirconia Ceramic Grinding Container .................................................................. 52 8507 Small Hardened Steel Grinding Container .......................................................... 52 8507R Rack .................................................................................................................... 548508 Small Tungsten Carbide Grinding Container ....................................................... 53 8521 Large Hardened Steel Grinding Container .......................................................... 53 8525 Transporter ......................................................................................................... 54 8530 SPEX SamplePrep 8530 Enclosed Shatterbox® ................................................... 50FFB SPEX Fusion Flux .................................................................................................. 72 FFB SPEX Fusion Flux Additives ................................................................................. 73 K1P Katanax® K1 Prime ............................................................................................. 64 K2P Katanax® K2 Prime ............................................................................................. 65 KP0010A Teflon® Beaker .................................................................................................... 68 KP0037A Mold Holder, for 30 mm molds ........................................................................... 68 KP0038A Mold Holder, for 32 mm molds ........................................................................... 68 KP0039A Mold Holder, for 35 mm molds ........................................................................... 68 KP0040A Mold Holder, for 40 mm molds ........................................................................... 68 KP0049A Zirconium Crucible .............................................................................................. 67 KP0235A Mold Plate, for 35 mm molds ............................................................................. 68 KP1001A Regular Crucible ................................................................................................. 66 KP1002A Heavy-Duty Crucible ........................................................................................... 66 KP1003A Mold – 30 mm ................................................................................................... 67 KP1004A Heavy-Duty Mold – 30 mm ................................................................................ 67 KP1005A Mold – 32 mm ................................................................................................... 67 KP1006A Heavy-Duty Mold – 32 mm ................................................................................ 67 KP1007A Mold – 35 mm ................................................................................................... 67 KP1008A Heavy-Duty Mold – 35 mm ................................................................................ 68 KP1009A Mold – 40 mm ................................................................................................... 68

6970M SPEX SamplePrep 6970EFM Enclosed Freezer/Mill®- Single Grinding Coil .......... 307151 Rack and Tongs ................................................................................................... 67 7151R Rack .................................................................................................................... 67 7151T Long Tongs ......................................................................................................... 677152 Graphite Crucibles .............................................................................................. 69 7152HP Graphite Crucibles .............................................................................................. 697153 Graphite Crucibles .............................................................................................. 69 7154 Short Tongs......................................................................................................... 67 7155 Graphite Crucibles .............................................................................................. 70 7156 Graphite Crucibles .............................................................................................. 70 7157 Graphite Crucibles .............................................................................................. 70 7158 Graphite Crucibles .............................................................................................. 71 7159 Graphite Crucibles .............................................................................................. 71 7160 Graphite Crucibles .............................................................................................. 71 7161 Graphite Crucibles .............................................................................................. 71 7162 Graphite Crucibles .............................................................................................. 72 8000D SPEX SamplePrep 8000D Dual Mixer/Mill........................................................... 40 8000M SPEX SamplePrep 8000M Mixer/Mill® ................................................................ 418001 Hardened Steel Vial Set ...................................................................................... 44 8001B Steel Ball Set ...................................................................................................... 488002 Polystyrene Vial .................................................................................................. 44 8003 Alumina Ceramic Vial Set .................................................................................... 45 8003A Alumina Ceramic Ball ......................................................................................... 488004 Tungsten Carbide Vial Set ................................................................................... 45 8004A Tungsten Carbide Ball ......................................................................................... 48 8004SS Steel-Jacketed Tungsten Carbide Vial Set ........................................................... 458005 Zirconia Ceramic Vial Set ..................................................................................... 45 8005A Zirconia Ceramic Ball .......................................................................................... 488006 Methacrylate Vial Set ......................................................................................... 458006A Methacrylate Balls .............................................................................................. 488007 Stainless Steel Vial Set ........................................................................................ 46 8007B Stainless Steel Ball Set ........................................................................................ 488008 Silicon Nitride Vial Set......................................................................................... 46 8008A Silicon Nitride Ball .............................................................................................. 498009 Round-Ended Hardened Steel Vial Set ................................................................ 46 8010 Multiple-Sample Adapter................................................................................... 49



KP1010A Heavy-Duty Mold – 40 mm ................................................................................ 68 KP1011A Platinum Crucible Heater Lid .............................................................................. 68 KP1015A Zirconium Crucible Heater Lid............................................................................. 68 KP5603A Mold holder, 30mm, 6-position.......................................................................... 68 KP5604A Mold holder, 32mm, 6-position.......................................................................... 68 KP5605A Mold holder, 35mm, 6-position.......................................................................... 68 KP5606A Mold holder, 40mm, 6-position.......................................................................... 68

Produc t Index

spex handbook 2011

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