versatile hybrid silica based (u)hplc columns the ... · pdf filethe selectivity company...

The Selectivity CompanyThe Selectivity Company

YMC-TriartVersatile hybrid silica based (U)HPLC columns

GENERAL

CATALOGuE

2013/2014

uHPLC

HPLC

BiO-LC

2 Introduction

Co., Ltd.YMC Karasuma-Gojo Bld. 284 Daigo-cho,Karasuma Nisiiru Gojo-dori Shimogyo-ku,Kyoto 600-8106 JapanTEL. +81(0)75-342-4515, FAX +81(0)75-342-4550www.ymc.co.jp

America, Inc.941 Marcon Boulevard Suite 201Allentown, PA18109 USATEL. +1-610-266-8650, FAX +1-610-266-8652www.ymcamerica.com

Europe GmbHSchöttmannshof 19D-46539 DinslakenGermanyTEL. +49(0)2064/427-0, FAX +49(0)2064/427-222www.ymc.de

India Ltd.CX - 07, 3rd Floor, Lobe - 1,A-Towers, The Corenthum, Plot No- A-41,Sector - 62, Noida - 201301 (U.P.) IndiaTEL. +91(0)120-4276020 - 25, FAX +91(0)120-4276026www.ymcindia.com

Co., Ltd. Shanghai Rep. OfficeFar East International Plaza A2404No. 319 Xianxia Road, Shanghai 200051P.R. ChinaTEL: +86-21-6235-1388, FAX: +86-21-6235-1398

Korea Co., Ltd.#208, Owners Tower, 16-5, Sunae-dong,Bundang-gu, Seongnam-si, Gyeonggi-do,463-825 KoreaTEL. +82-31-716-1631, FAX +82-31-716-1630www.ymckorea.com

If any of your requirements for selecting a chromatography vendor are not addressed in this general catalogue, please contact “your” YMC office below. You will find that YMC is always highly respon-sive to customer requirements, and feedback is guaranteed.

Taiwan Co., Ltd.3F, No. 1353, Zhongzheng Rd.,Taoyuan City, Taoyuan Country 330,Taiwan (R.O.C.)TEL. +886-3-2150-630, FAX +886-3-2150-286www.ymctaiwan.com

Worldwide Availability

Introduction 3

* Application data by courtesy YMC Co., Ltd.

IntroductionCompany Profile ........................................................ 8-9YMC Websites ........................................................ 10-11

ProFamilyIntroduction ................................................................ 51YMC UltraHT Pro C18 / Hydrosphere C18 ............ 52-59General .................................................................. 60-67YMC-Pack Pro C18 ............................................... 68-69YMC-Pack Pro C8 ................................................. 70-71YMC-Pack Pro C4 ................................................. 72-73YMC-Pack Pro C18 RS ......................................... 74-77Hydrosphere C18 ................................................... 78-79Ordering Information .............................................. 80-81

Reversed Phase “Classics”Introduction ................................................................. 83Products .............................................................. 84-107YMC-Pack ODS-AQ, ODS-A, ODS-AM, ODS-AL,PolymerC18, YMCbasic, YMC-Pack C8 (Octyl),Ph (Phenyl), C4 (Butyl), TMS (C1), CN (Cyano)Ordering Information ........................................... 108-110

Normal Phase ChemistriesIntroduction ................................................................ 113Products ............................................................ 114-125YMC-Pack SIL (Silica), PVA-Sil, CN (Cyano),Diol, Polyamine II, NH2 (Amino), TMS (C1)Ordering Information ........................................... 126-127

BiochromatographyIntroduction ................................................................ 129Selection Guide .................................................. 130-131Ion Exchange (IEX) ............................................ 132-141YMC-BioPro QA/SP, YMC-BioPro QA-F/SP-FOrdering Information ........................................... 141-142Size Exclusion (SEC) ......................................... 143-154YMC-Pack DiolOrdering Information .................................................. 155Reversed Phase (RP) ........................................ 157-164YMC-Triart C18, YMC-Pack Pro C18, ODS-A,ODS-AQ, Hydrosphere C18Ordering Information ........................................... 165-166Glass Columns .................................................. 167-170ECO, ECOPLUS, PilotOrdering Information ........................................... 171-175

Chiral ColumnsIntroduction ................................................................ 177YMC Chiral NEA(R)(S) ......................................... 178-181YMC Chiral CD BR .............................................. 182-185Ordering Information .................................................. 186

Speciality ColumnsIntroduction ................................................................ 189YMC30 / YMC Carotenoid .................................. 190-191YMC PAH ........................................................... 192-193J’sphere ODS ..................................................... 194-199Ordering Information ................................................. 200

MiscellaneousTechnical Information / Introduction .......................... 217Column Handling ................................................ 218-220Mobile Phases for RP-Columns ................................ 221HPLC Column Performance ...................................... 222Inspection Reports .................................................... 223FAQ .................................................................... 224-227Troubleshooting .................................................. 228-231Basic Data ................................................................. 232Solvent Miscibility Table ............................................ 233Column Selection Guide ..................................... 234-235Linear Scale Up ......................................................... 236Preparative Column Selection Guide ........................ 237Substance Index ................................................. 238-246Full Listing of all Chemistries and Dimensions .......... 247

YMC-Actus SEMI-PREPIntroduction ................................................................. 43Stability ....................................................................... 44Axial Compression Technology .................................... 45Application ............................................................. 46-47Ordering Information ............................................... 48-49

YMC-Triart HYBRIDIntroduction ................................................................. 13First Choice / Particle Technology .............................. 14pH & Temperature Stability ......................................... 15Method Transfer between HPLC UHPLC .......... 16-17UHPLC ................................................................... 18-21LC/MS ................................................................... 22-24Pharmaceuticals .................................................... 25-29Food ....................................................................... 30-31Peptides ....................................................................... 31HILIC ...................................................................... 32-33QC Data ................................................................. 34-39Ordering Information ................................................... 40

➞

➞

FlashIntroduction ............................................................... 203Advantages ............................................................... 204Application / Product Finder ..................................... 205Ordering Information .......................................... 206-207

Contents

Preparative & Process LCIntroduction .............................................................. 209General .............................................................. 210-211YMC Silica(s) ............................................................ 212YMC BioPro - IEX Ion Exchange Media .................... 213YMC-Triart ................................................................. 214

What’s new? .............................................. 5

4 Introduction

UniversalYMC-Triart

for acidic, basic andneutral analytes

TransferScalable particles:

EASYUHPLC ↔ HPLC Flexible

YMC-Triart:pH 1-12

Temperaturesup to 70°C


YMC-TriartVersatile hybrid silica based (U)HPLC columns

www.ymc.de


YMC*Gel HG-seriesHigh grade silica phases for preparative HPLC

www.ymc.de

Available Literature (Selection)

IEXSECRP

NP/HILIC

YMC Phases forBiochromatography

UltraFastHPLC

Application Guide

The Selectivity Company

YMC-DispoPackAT

Premium Flash Cartridges

Schöttmannshof 1946539 Dinslaken

Phone: +49 (0) 2064 427-0Fax: +49 (0) 2064 427-222

E-Mail: [email protected]

www.ymc.de


YMC-TriartScaleable and pH stable hybrid silica process material

www.ymc.de

Multilayerhybrid process

material Stableat high pH -

increased CIP-stability

Innovativemicro-reactor

technology

Analytical Preparative

PrepBulk

media

YMC-BioProIEX materials

Also available:

Posters

Product Information

Technical Guide Lines

Manuals

DMF / RSF / CoA

etc.

Introduction 5


What’s new?

Versatile (U)HPLC-hybrid columnsWith YMC-Triart challenging pH-stability and high temperatures are no longer a limitation to your work.

YMC-Triart C18, C8 and HILIC-Diol in their particle sizes of 1.9 µm, 3 µm, 5 µm or 10 µm combined with various column dimensions makes it suitable for any LC-equipment: (U)HPLC, semi-preparative to process scale bulk. Process scale particle sizes are available in multi-ton scale.

UniversalYMC-Triart




YMC-Triart:pH 1-12


page

13

page

43

page

203

YMC-Actus for fast semi-preparative HPLC high throughput separations Semi-preparative chromatography is the link between analytical HPLC and preparative LC. Even though the chromatographic systems used for semi-preparative LC do not reach the size of preparative LC systems, the objectives remain the same:

• Purificationandisolationofmaximumsamplequantity

• Savingsintimeandcosts.

With YMC-Actus time is on your side!

YMC-DispoPackAT Flash Cartridges - highquality products for your purification successBy using the new cartridges with 25 µm spherical silica you are able to obtain higher resolution in less than half of the time, compared to classical 40/63 irregular particles.

YMC-DispoPackAT is available in sizes ranging from 12 g to 800 g and in following functional groups: SIL (silica gel), NH2 (amino), Diol, and ODS (C18).

page

238

Substance IndexIn order to facilitate the search for applications, the substance index contains 1091 substances and it will provide reference to the cor-responding page in the catalogue or in the ”YMC Application Data Collections ”, respectively.

6 Introduction

[Kinkalku-ji] japanese

Kinkaku-ji (Temple of the Golden Pavilion), also known as Rokuon-ji (Deer Garden Temple), is a ZenBuddhist temple in Kyoto, Japan. The garden complex is an excellent example of Muromachi period garden

design. The Muromachi period is considered to be a classical age of Japanese garden design. Thecorrelation between buildings and its settings were greatly emphasized during this period. It was a way to integrate the structure within the landscape in an artistic way. The garden designs were characterized by

a reduction in scale, a more central purpose, and a distinct setting. A minimalistic approach was brought to the garden design, by recreating larger landscapes in a smaller scale around a structure.

It is designated as a National Special Historic Site and a National Special Landscape, and it is one of 17locations comprising the Historic Monuments of Ancient Kyoto World Heritage Site. It is also one of the

most popular buildings in Japan, attracting a large number of visitors annually.

Introduction 7


8 Introduction

Company ProfileYMC is a leading specialist supplier of high performance products for liquid chromatography, with headquarters in Kyoto, Japan, and with subsidiaries in the USA, in India, China, Korea, Taiwan and Europe.

As a mission statement, it is our ambition to provide chromatographic solutions for any compound from its discovery through scale-up into production and its ultimate quality control in the laboratory. YMC can always be expected to provide meaningful support, chromatographic tools and assistance for routine R&D, fast LC/UHPLC, high-throughput screening, LC-MS, automation or process-scale engineering.

YMC Co., Ltd. manufacturing facility in Komatsu, Japan

Today, YMC has developed beyond their traditional, yet still high performance silica-based materials to produce three new, distinct high technology platforms:

• anewgenerationofsilica-basedmaterialsforprocess chromatography, YMC HG-series, with enhanced physical and chemical properties

•amethacrylate-based,porousandnon-porousbead for ion exchange chromatography (IEX) from analytical to process scale with particle sizes from 5 to 75 µm

• YMC-Triart,anewhybrid-styleorganic/inorganicparticle, a “global first” which provides a fully scalable series of products from analytical to process scale. Available in up to multi-ton capacity per year and with major benefits regarding mechanical/chemical stability which result in enhanced column lifetime.

However, it is not only product specification that demonstrates YMC qualities, but also the perceived ethical values of having people willingly contribute competent and consistent performance along with exceptional reliability even for difficult and demanding separations. YMC strive to exceed expectations – day after day, year after year – with worldwide availability and with guaranteed long-term supply.

YMC Europe GmbH, Dinslaken, Germany

Introduction 9


Company ProfileHands-on support services are offered by Ko-matsu/Japan, Allentown/PA, USA and Dinslak-en/Germany: local application laboratories and the Komatsu factory provide method develop-ment, optimisation and scale-up as well as custom synthesis and toll manufacturing from milligram to ton scale.

Facilities include preparative HPLC, fraction concentration equipment, crystallisation, vac-uum filtration, vacuum drying, freeze drying, all supported by state-of-the-art analytical instru-mentation. The “live experience” by YMC spe-cialists added value aspect to make customers successful in their work: for robust, valid and fast methods in the lab, with high sensitivity, and increased output. YMC training courses are available with a well-defined balance of theory and practical work at either novice or advanced levels, all of which can be customised to indi-vidual requirements.

Supply

Extensive local inventory and highly motivated order processing staff ensure speedy product supply to virtually any destination. Individual items are stocked based on statistical consump-tion or customised as specified by purchasing officers or procurement agreements in place for just-in-time supply. In addition, authorized YMC International Distributors are encouraged to maintain local inventory, too, occasionally complemented by consignment products pro-vided by YMC, so that lead times are brought down to a minimum.

10 Introduction

YMC Websites- more than 700 pages full of useful information

- detailed description of more than 80 stationary phases

- extensive applications database

- latest news at a glance on the homepage

Although we have put all our efforts into providing you with the best information on the successful implementation of YMC products for liquid chromatography in this catalogue, printed documents by nature remain unidirectional media compared with lively internet platforms.

www.ymc.co.jp

for Asia

www.ymcamerica.com

for the Americas

www.ymc.de

for EMEA Countries

Introduction 11


YMC Website “www.ymc.de”

“Contact”:

If you just want to know the “classical” contact information including phone number, email and postal address.

“News”:

Get all the latest information and be up-to-date by choosing www.ymc.de. YMC continuously de-velops new products to facilitate the routine in laboratories. In addition new brochures, applications and other support can be found at a glance and can be downloaded immediately. You will also be able to read our latest YMC Newsletter or browse in our news archive.

“Download area”:

You will find applications, brochures, publications, support information and much more in pdf-file format in our download area.

“Online Communication Platform - OCP”:Our online communication platform is a new and unique alternative

way to contact YMC Europe in addition to phone or email. You can askour product specialists for technical support or you can be transferred

to our sales administration team if you have a supply question.Try out the next generation of online customer support.

Instead of simply reproducing this catalogue in an internet format, you will find that www.ymc.de incorporates many additional features: updates, various commonly used contact information, down-loads, search and useful links, an application database as well as taking the challenge with our new “YMC Online Communication Platform”. OCP will bring you – online – to the desk of an experienced product specialist for your personal, direct access to online support, which can include technical questions, requests for specific application methods or “simple questions” such as local supply facilities or product information. Visit us at www.ymc.de.

12 YMC-Triart

UniversalYMC-Triart




YMC-Triart:pH 1-12



YMC-Triart 13

Contents

First Choice / Particle Technology ........................................ page 14

pH & Temperature Stability ................................................... page 15

Method Transfer between HPLC UHPLC ..................... page 16-17

UHPLC ............................................................................ page 18-21

LC/MS .............................................................................. page 22-24

Pharmaceuticals ............................................................. page 25-29

Food ................................................................................ page 30-31

Peptides ............................................................................... page 31

Diol-HILIC ........................................................................ page 32-33

QC Data ...........................................................................page 34-39

Ordering Information ............................................................. page 40

YMC-Triart

Chromatographers always seek to push the limits of HPLC columns to greater extremes to allow them to perform day-to-day with ever-changing pH, buffers and temperature ranges. The column for the laboratory of today must be suitable for harsh pH conditions in combination with high temperature ranges without sacrificing selectivity. In addition narrow, symmetrical peak shapes are necessary in order to cope with rapid analysis of demanding samples. This has required manufacturers to seek more innovative ways to produce suitable stationary phases.

In order to meet these goals, YMC has developed a new particle technology. This is based on a multi-layered particle produced via a tightly controlled granulation technology which has been adapted from micro-reactor technology. The revolutionary production technique provides a multi-layer silica-organic hybrid stationary phase, which provides an outstandingly narrow pore size and particle size distribution. This in turn, results in low back pressures and high load-ability.

Introduction

➞➞

14 YMC-Triart

Particle technology

YMC-Triart is a multi-layered material prepared using tightly controlled particle formation tech-nology which has been adapted from micro-reactor technology. This recently developed production process results in exceptionally narrow particle and pore size distributions.

With YMC-Triart, challenging pH and high tem-perature conditions are no longer a limitation to the day-to-day work in laboratories. Most im-portantly, due to its unique particle composition, a balanced hydrophobicity and silanol activity are achieved which makes YMC-Triart a ”First Choice“ column in method development.

inorganic siloxane layer

organic siloxane layer

YMC-Triart hybrid structure

Specification YMC-Triart C18 YMC-Triart C8 YMC-Triart Diol-HILIC

Baseorganic/inorganic

silicaorganic/inorganic

silicaorganic/inorganic

silica

Stationary phase C18 (as USP L1) C8 (as USP L7) Diol (as USP L20)

Particle size 1.9, 3 and 5 µm 1.9, 3 and 5 µm 1.9, 3 and 5 µm

Pore size 12 nm 12 nm 12 nm

Bonding polymeric type polymeric type —

End-cappingmulti-stageendcapping

multi-stageendcapping

—

pH range 1 ~ 12 1 ~ 12 2 ~ 10

Temperature range(upper limit)

pH 1-7: 70 °C,pH 7-12: 50 °C

pH 1-7: 70 °C,pH 7-12: 50 °C

50 °C

Conventional hybrid silica-based ODS columns tend to be less hydrophobic than silica-based columns. YMC-Triart C18 has a higher carbon load, giving it a hydrophobicity comparable to that of standard ODS columns, thereby making it a ”versatile first-choice“ column for method develop-ment.

Hydrophobicity k‘ (amylbenene)

Hyd

roge

n bo

ndin

g ca

paci

tya

(caff

eine

/ben

zene

)

Standard ODSThe most versatilecolumns as first-choice.

First choice column for method development

Triart C18Symmetry C18

ZORBAX Extend-C18

Triart C8


YMC-Triart 15

0 10 20 30 40 50

YMC-Triart C18

0%

20%

40%

60%

80%

100%

120%

Hybrid-silica based C18Silica based C18

Versatile wide pH stability

Triethylamine (pH 11.5, 40 °C)*Phosphate buffer (pH 11.5, 40 °C)*

Column: 5 µm, 150 x 4.6 mm IDPart No.: TA12S05-1546WTEluent: 50 mM K2HPO4-K3PO4 (pH 11.5) / methanol (90/10)Flow rate: 1.0 ml/minTemperature: 40 °CSample: benzyl alcohol

0 300250200150100500%

20%

40%

60%

80%

100%

120%

YMC-Triart C18

Silica based C18

Time (hr)

Rat

e of

theo

retic

al p

late

num

ber

Column: 5 µm, 150 x 4.6 mm IDPart No.: TA12S05-1546WTEluent: 50 mM triethylamine (pH 11.5) / methanol (90/10)Flow rate: 1.0 ml/minTemperature: 40 °CSample: benzyl alcohol

Stability at high temperature

pH 1, 70 °C*pH 6.9, 70 °C*

Column: 5 µm, 50 x 2.0 mm IDPart No.: TA12S05-0502WTEluent: 20 mM KH2PO4-K2HPO4 (pH 6.9) / acetonitrile (90/10)Flow rate: 0.2 ml/minTemperature: 70 °CSample: phenol Column: 5 µm, 50 x 2.0 mm ID

Part No.: TA12S05-0502WTEluent: acetonitrile / water (60/40)Flow rate: 0.2 ml/minTemperature: 70 °CSample: butyl benzoate

YMC-Triart phases show great chemical stability due to the newly developed hybrid-silica. Even under high pH or high temperature conditions, the lifetime of YMC-Triart phases is more than 10x greater than conventional reversed phase columns.

0%

20%

40%

60%

80%

100%

120%

0 100 200 300 400 500 600 700

YMC-Triart C18

Silica based C18alkaline-resistant type

Silica based C18

0 120100806040200%

20%

40%

60%

80%

100%

120%

YMC-Triart C18

Silica based C18

Hybrid-silica based C18

Time (hr)

Rat

e of

theo

retic

al p

late

num

ber

The columns are stored in acetonitrile/water/TFA (10/90/1) 70 °C, and the performance tested every 20 hours.

YMC-Triart C18 YMC-Triart C18YMC-Triart C8

YMC-Triart C18YMC-Triart C8

YMC-Triart C18

pH & temperatureR

ate

on

N [%

]

Time [h]

Rat

e o

n N

[%]

Time [h]

16 YMC-Triart

Transfer HPLC UHPLC

YMC-Triart C183 µm TF(2) 1.43 1.9 µm TF(2) 1.47

X-Bridge BEH C18 and Acquity UPLC BEH C182.5 µm TF(2) 2.24 1.7 µm TF(2) 2.35

KinetexTM C182.6 µm TF(2) 6.49 1.7 µm TF(2) 4.94

Column: 50 x 2.0 mm ID or 2.1 mm IDEluent: 20 mM KH2PO4-K2HPO4 (pH 6.9) / acetonitrile (65/35)Temperature: 40 °CFlow rate: 0.2 ml/minDetection: UV at 235 nm

1. Chlorpheniramine (basic)2. Dextromethorphan (basic)3. Propyl paraben (internal standard)

Case Studies**

Secure your method transfer!

Differences in selectivity, retention time, and also peak shapes between different particle sizes of commercially available C18 phases in the same brand (or an alternative as recommended by its manufacture) have been observed.

YMC has adressed this issue of method transfer. YMC-Triart columns show identical selectivity and excellent peak shapes for basic compounds for all 3.0 µm to 1.9 µm particle sizes. It allows predictable scale up from UHPLC to conventional HPLC and even to semi-preparative LC, and vice versa.

KinetexTM C18 columns show significant peak tailing and have limited scalability due to lack of larger particle sizes.

** These observations might not be representative for all applications but have been reported in some cases.

➞➞


YMC-Triart 17

Evaluation of method transfer performance!*

YMC-Triart C18150 x 2.0 mm ID, 5 µmTA12S05-1502WT4.5 MPa (650 psi)

YMC-Triart C1850 x 2.0 mm ID, 1.9 µmTA12SP9-0502PT37 MPa (5400 psi)

5% B (0-3 min)5-30% B (3-10 min)30-45% B (10-55 min)0.2 ml/min

5% B (0-0.3 min)5-30% B (0.3-1.0 min)30-45% B (1.0-6.0 min)0.6 ml/min

Eluent: A) water/ formic acid (100/0.1) B) acetonitrile/ formic acid (100/0.1)Temperature: 40 °CDetection: UV at 240 nmInjection: 1 µl (5 µg/ml)

55 min

6 min

1. Oxine-copper (Copper 8-quinolinolate)2. Asulam3. Thiram4. Triclopyr5. Mecoprop6. Flazasulfuron7. Siduron8. Halosulfuron- methyl9. Azoxystrobin

Method transfer between HPLC UHPLC*

HypersilGOLD5 µm

XBridge C18 5 µm

L-column2 ODS5 µm

Triart C183 µm 1.9 µm

Kinetex C18 2.6 µm

Kinetex C18 1.7 µm

ACQUITY UPLC BEH C18 1.7 µm

Hypersil GOLD1.9 µm

L-column2 ODS2.0 µm

Hydrophobicity [k‘ (Amylbenzene)]

Hyd

rog

en b

ond

ing

cap

acit

y[a

(C

affe

ine

/Ben

zene

)]

0.06

0.08

0.10

0.12

2 4 6 8

Eluent: methanol/water (80/20) [Amylbenzene] methanol/water (30/70) [Caffeine, Benzene]

With the introduction of UHPLC, sub-2-µm particles became neccessary. Therefore smaller par-ticles have been added to existing column lines. Consequently, sub-2-µm particles may exhibit differences in chromatographic performance.By introducing YMC-Triart, YMC provides matching chromatographic behaviour for all particles sizes!

Transfer HPLC UHPLC➞➞

➞➞

18 YMC-Triart

UHPLC

Column: 50 x 2.0 or 2.1 mm IDEluent: 20 mM KH2PO4-K2HPO4 (pH 6.9)/ acetonitrile (65/35)Flow rate: 0.2 ml/minDetection: UV at 235 nmTemperature: 40 °C

TF(2)=1.47

TF(2)=5.05

TF(2)=6.49

TF(2)=4.94

1 2 3

1 23

12,3

12,3

1.9 µm

2.7 µm

2.6 µm

1.7 µm

YMC-Triart C18

Ascentis Express C18

Kinetex C18

Higher resolution and good loadability*

Lower HETP means higher resolution!*

Column: 50 x 2.0 or 2.1 mm IDEluent: acetonitrile / water (60/40)Detection: UV at 254 nmSample: Butyl benzoate

0,015

0,010

0,005

0,000

HE

TP

(m

m/p

late

s)

0 2 4 6 8 10 12

Linear velocity (mm/sec)

2.7 µmAscentis Express(Core-shell type)

2.6 µmKinetex C18(Core-shell type)

1.7 µmKinetex C18 (Core-shell type)

1.9 µmYMC-Triart C18


YMC-Triart 19

UHPLC & MS

Determination of Artificial Sweeteners with LC-MS/MS

Acesulfam-K Saccharin Cyclamat-Na

Non-biological markers of wastewater entries in ground and surface water

Extracted Ion Chromatogram (XIC) of Acesulfam-K, 0.1 µg/L Extracted Ion Chromatogram (XIC) of Saccharin, 0.1 µg/L

Extracted Ion Chromatogram (XIC) of Cyclamat-Na, 0.1 µg/L

Column: YMC-Triart C18, 12 nm, 1.9 µm, 100 x 3.0 mm IDPart No.: TA12SP9-1003PTLC-System: Agilent 1100 HPLC system and CTC Analytics HTC-Pal AutosamplerMS/MS System: Applied Biosystems MDS Sciex API 4000, ESI negativeTemperature: 35°CFlow rate: 0.3 ml/minInjection: 40 µL, direct injectionEluent: A: H2O (containing 10 mmol NH4 formate) B: MeOH (containing 10 mmol NH4 formate)Gradient: Time 0 6.0 6.1 12.0 % B 2 75 2 2

by courtesy of: Thomas Class, Sandro JooßPTRL Europe, Helmoholtzstraße 22, Science Park I, D-89081 Ulm

20 YMC-Triart

UHPLC

Angiotensin I, II and III* Insulin*

Column: YMC-Trairt C18, 12 nm, 1.9 µm, 50 x 2.0 mm IDPart No.: TA12SP9-0502PTEluent: A) H2O + 0.1% TFA B) acetonitrile + 0.1% TFAGradient: 30% B (0 min); 30-32% B (0-5 min); 32% B (55 min)Flow rate: 0.6 ml/minDetection: UV at 220 nmPressure: 611 barInjection: 0.5 µlTemperature: 30 °C

1. Angiotensin I2. Angiotensin II3. Angiotensin III

1. Bovine insulin2. Human insulin3. Porcine insulin

Macrolide antibiotics*

Column: YMC-Trairt C18, 12 nm, 1.9 µm, 50 x 2.0 mm IDPart No.: TA12SP9-0502PTEluent: A) 20 mM K2HPO4 + 20 mM KH2PO4 (pH 7.9) B) acetonitrileGradient: 60% B (0.5 min); 60-70% B (0.5-1.5 min); 70% B (3.5 min)Flow rate: 0.45 ml/minDetection: UV at 210 nmPressure: 520 barInjection: 1 µlTemperature: 50 °C

1. Erythromycin2. Spiramycin

Sulpha drugs*

Column: YMC-Trairt C18, 12 nm, 1.9 µm, 50 x 2.0 mm IDPart No.: TA12SP9-0502PTEluent: H2O + formic acid (pH 2.5) / acetonitrile (75/25)Flow rate: 0.75 ml/minDetection: UV at 280 nmPressure: 740 barInjection: 0.5 µlTemperature: 50 °C

1. Uracil2. Sulphathiazole3. Sulphamerazin4. Sulphamethaxole

1

2

3

1

23

1

2

1

2

3

4

Column: YMC-Trairt C18, 12 nm, 1.9 µm, 50 x 2.0 mm IDPart No.: TA12SP9-0502PTEluent: 20 mM KH2PO4 + K2HPO4 (pH 7.9) / acetonitrile (22/78)Flow rate: 0.7 ml/minDetection: UV at 220 nmPressure: 720 barInjection: 0.5 µlTemperature: 40 °C


YMC-Triart 21

UHPLC

Tetracycline antibiotics*

Eluent: A) water / HFBA* (100/0.1) B) acetonitrile / HFBA* (100/0.1)Temperature: 37 °CDetection: UV at 290 nmInjection: 1 µl (0.1 mg/ml)*heptafluorobutyric acid

Fast LC for conventional HPLC*

optimisation

Down scaling

YMC-Triart C18, 5 µm, 150 x 2.0 mm ID

1. Oxytetracycline2. Tetracycline3. Chlortetracycline

Betablockers*

1. Atenolol2. Pindolol3. Nadolol4. Metoprolol5. Propranolol

3

2

1

4 5

3

2

1

Column: YMC-Trairt C18, 12 nm, 1.9 µm, 50 x 2.0 mm IDPart No.: TA12SP9-0502PTEluent: 5 mM CH3COONH4 / acetonitrile (87/13)Flow rate: 0.65 ml/minDetection: UV at 280 nmPressure: 662 barInjection: 1 µlTemperature: 40 °C

Column: YMC-Trairt C18, 12 nm, 1.9 µm, 50 x 2.0 mm IDPart No.: TA12SP9-0502PTEluent: A) 20 mM CH3COONH4 + ammonia (pH 9.0) B) acetonitrileGradient: 25% B (1.0 min); 75% B (1-6 min)Flow rate: 0.35 ml/minDetection: UV at 254 nmPressure: 450 barInjection: 1 µlTemperature: 40 °C

Baseline separation,but long analysis time

insufficientseparation

Baseline separation,with short time

YMC-Triart C18, 3 µm, 50 x 2.0 mm ID

5% B, 0.4 ml/min,12.2 MPa

5% B, 0.2 ml/min, 5.6 MPa10% B, 0.2 ml/min, 5.7 MPa

22 YMC-Triart

LC/MS

LC/MS compatibility*

Column: 5 µm, 50 x 2.0 mm IDPart No.: TA12S05-0502WTEluent: A) water / formic acid (100/0.1) B) acetonitrile / formic acid (100/0.1) 5% B (0-1 min), 5-100% B (1-5 min), 100% B (5-10 min), 100-5% B (10-10.1 min), 5% B (10.1-12.5 min)Flow rate: 0.4 ml/minTemperature: 40 °CDetection: ESI positive, TIC (Mass Range: 50-1000)

Column bleeding, caused by the fragments of stationary phase, is the main reason for background noise and restric-tions on detection limits. No bleed is observed in the test of total ion current (TIC) measured by LC/MS with blank or with YMC-Triart C18. So in terms of the signal/noise ratio (S/N ratio), YMC-Triart C18 can be expect to not only reduce the background noise but to also increase the sensitivity of the analysis.

— YMC-Triart C18

— No column

LC/MS analysis of benzodiazepine derivates*

Column: YMC-Triart C18 (5 µm, 12 nm) 50 x 2.0 mm IDPart No.: TA12S05-0502WTEluent: A) 10 mM formic acid B) acetonitrileGradient: 25-50% B (0-10 min)Flow rate: 0.2 ml/minTemperature: 40 °CDetection: Bruker Daltonics micrOTOF, ESI, positive modeInjection: 5 µl (100 ng/ml)

Peak 1: 100ng_ml_RB4_01_2291.d: EIC 343.051±0.01+All MSPeak 2: 100ng_ml_RB4_01_2291.d: EIC 343.078±0.01+All MS

Courtesy of J. Watanabe, Bruker Daltonics K. K.


YMC-Triart 23

LC/MS

LC/MS analysis of tetracycline antibiotics*

Column: YMC-Triart C18 (5 µm, 12 nm) 50 x 2.0 mm IDPart No.: TA12S05-0502WTEluent: acetonitrile / water / formic acid (15/85/0.1)Flow rate: 0.4 ml/minTemperature: 40 °CDetection: A) UV at 270 nm B) ESI positive-modeInjection: 10 µl (1 µg/ml)

24 YMC-Triart

LC/MS

Analysis of 360 pesticides in a single run

Column: YMC-Triart C18 (3 µm, 100 x 2.0 mm ID)Part No.: TA12S03-1002WTEluent: A) 5 mM ammonium formate / water B) 5 mM ammonium formate / methanolFlow rate: 0.25 ml/minTemperature: 45 °C

Injection: 5 µlGradient: 0 min: 30% B, 0.1 min: 50% B, 18 min: 100% B, 21 min: 100% B, 21.01 min: 30% B, 29 min: 30% BTotal run time: 30 minSample: 100 ng/ml pesticide mix in acetonitrile

by courtesy of: József LászlóWIREC, WESSLING International Research and Educational Centre Nonprofit Co. (Hungary)


YMC-Triart 25

Separation of alkaloids*

Column: YMC-Triart C18 (5 µm, 12 nm) 50 x 2.0 mm IDPart No.: TA12S05-0502WTEluent: 20 mM CH3COOH-CH3COONH4 (pH 4.9) / acetonitrile (80/20)Flow rate: 0.2 ml/minTemperature: 40 °CDetection: UV at 220 nmInjection: 1 µl (0.02-0.1 mg/ml)

1. Scopolamine2. Atropine3. Cinchonine4. Quinine5. Dihydroquinine

Barbiturates in human serum*

Column: YMC-Triart C18 (5 µm, 12 nm) 50 x 2.0 mm IDPart No.: TA12S05-0502WTEluent: A) 20 mM HCOONH4-NH3 (pH 9.5) B) methanolGradient: 0-90% B (0-7 min)Flow rate: 0.2 ml/minTemperature: 25 °CDetection: UV at 240 nmInjection: 1 µl

A) Standard (10 µg/ml)

B) Spiked human serum

Solid-phase extraction methodYMC Dispo SPE C18 100 mg/1ml

Condition2 ml methanol

2 ml water

Load500 µl spiked human serum

solution (each 10 µg)

Elute500 µl methanol/water (85/15)

Dilute500 µl 20 mM ammonium

formate buffer (pH 9.5)

Pharmaceuticals

26 YMC-Triart

Pharmaceuticals

Erythromycin at elevated pH and temperature*

Column: YMC-Triart C18 (3 µm, 12 nm)Dimension: 50 x 2.0 mm IDPart No.: TA12S03-0502WTEluent: 20 mM KH2PO4-K2HPO4 / acetonitrile / methanol (40/45/15)Flow rate: 0.2 ml/minDetection: UV at 210 nm

1. Optimisation of pH

2. Optimisation of temperature (pH 7.9)

3. Stability test: pH 7.9, 70 °C

Num

berof injections


YMC-Triart 27

Pharmaceuticals

Column: YMC-Triart C18 (5 µm, 12 nm)Dimension: 150 x 3.0 mm IDPart No.: TA12S05-1503WTEluent: acetonitrile / 10 mM H3PO4 (30/70)Flow rate: 0.425 ml/minTemperature: 37 °CDetection: UV at 280 nmInjection: 2 µl (50 µg/ml)

Separation of flavonoids*

Column: YMC-Triart C18 (5 µm, 12 nm)Dimension: 150 x 3.0 mm IDPart No.: TA12S05-1503WTEluent: 10 mM CH3COOH-CH3COONH4 (pH 4.2) / acetonitrile (75/25)Flow rate: 0.425 ml/minTemperature: 40 °CDetection: UV at 254 nmInjection: 4 µl (0.02 ~ 0.3 mg/ml)

Separation of aromatic carboxylic acids*

28 YMC-Triart

Pharmaceuticals

Aciclovir syrup and injection*

Column: YMC-Triart C18 (5 µm, 12 nm) 150 x 4.6 mm IDPart No.: TA12S05-1546WTEluent: phosphate buffer* / methanol (95/5) *Dissolve 1.45 g of H3PO4 and 25 ml of 1 mol/l CH3COOH in water to make 900 ml q adjust pH 2.5 by 1 mol/l NaOH q add water to make 1000 mlFlow rate: 1.0 ml/minTemperature: 25 °CDetection: UV at 254 nmInjection: 20 µl (0.05 mg/ml, 0.032 mg/ml)

Column: YMC-Triart C18 (5 µm, 12 nm) 250 x 4.6 mm IDPart No.: TA12S05-2546WTEluent: 50 mM KH2PO4 (pH 7.5 adjusted by 8 M KOH) / acetonitrile (55/45)Flow rate: 1.0 ml/minTemperature: 25 °CDetection: UV at 210 nmInjection: 10 µl

Clindamycin hydrochloride*

Column: YMC-Triart C18 (5 µm, 12 nm) 150 x 3.0 mm IDPart No.: TA12S05-1503WTEluent: 10 mM CH3COOH-CH3COONH4 (pH 4.2) / acetonitrile (75/25)Flow rate: 0.425 ml/minTemperature: 40 °CDetection: UV at 254 nmInjection: 4 µl (0.02 ~ 0.3 mg/ml)

Analysis of amlodipine besilate*


YMC-Triart 29

Pharmaceuticals

Basic drugs*

1. Hydrochlorothiazide2. Amlodipine besilate3. Valsartan4. Atorvastatin calcium hydrate5. Candesartan cilexetil

Sequential analysis of Carvedilol*

Column: YMC-Triart C8 (5 µm, 150 x 2.0 mm ID)Part No.: TO12S05-1502WTEluent: phosphate buffer (pH 2.0)* / acetonitrile (65/35) *Dissolve 2.72 g of KH2PO4 in 900 ml water, adjust pH 2.0 with H3PO4 and add water to make 1000 mlFlow rate: 0.28 ml/min (adjust the flow rate so that the retention time of carvedilol is about 4 min)Temperature: 55 °CDetection: UV at 240 nm

No change in retention time is observed even under a high pH and at a elevated temperature.

YMC-Triart C8

YMC-Triart C8

Column: YMC-Triart C8 (3 µm, 12 nm), 50 x 2.0 mm IDPart No.: TO12S03-0502WTEluent: A) water / formic acid (100/0.1) B) acetonitrile / formic acid (100/0.1) 10-90% B (0-5 min), 90% B (5-7 min)

Flow rate: 0.4 ml/minTemperature: 30 °CDetection: UV at 254 nmInjection: 2 µl (10-20 µg/ml)

30 YMC-Triart

Food

Separation of water-soluble vitamins*

Column: YMC-Triart C18 (5 µm, 12 nm) 250 x 4.6 mm IDPart No.: TA12S05-2546WTEluent: phospate buffer* / acetonitrile (90/10) * Dissolve 1.4 g KH2PO4 in 800 ml water q add 26 ml 10% TBA·OH q adjust pH 5.2 by 20% H3PO4 q add water to make 1000 mlFlow rate: 0.8 ml/minTemperature: 40 °CDetection: UV at 260 nmInjection: 10 µl (5 µg/ml)

1. Thiamine HCl (Vitamin B1)2. Pyridoxine HCl (Vitamin B6)3. Nicotinamide4. Cyanocobalamin (Vitamin B12)5. L-Ascorbic acid 2-glucoside6. L-Ascorbic acid (Vitamin C)7. Erythorbic acid8. Riboflavin (Vitamin B2)9. Nicotinic acid

Column: YMC-Triart C18 (3 µm, 12 nm) 50 x 2.0 mm IDPart No.: TA12S03-0502WTEluent: A) acetonitrile / water / HCOOH (10/90/0.1) B) acetonitrile / water / HCOOH (60/40/0.1)

Soy isoflavones in supplement*

A) Standard (0.01 mg/ml)

B) Supplement (1 g/50 ml)

1. Daidzin 2. Glycitin 3. Genistin 4. 6“-O-Malonyldaidzin 5. 6“-O-Malonylglycitin 6. 6“-O-Acetyldaidzin 7. 6“-O-Acetylglycitin 8. 6“-O-Malonylgenistin 9. Daidzein 10. Glycitein 11. 6“-O-Acetylgenistin 12. Genistein

Sample preparation method

Supplement

Stirred at r.t. for 1 hr

Filtration

Injection

50% aqueous ethanol

Gradient: 5-40% B (0-12 min)Flow rate: 0.4 ml/minTemperature: 25 °CDetection: UV at 254 nmInjection: 2 µl


YMC-Triart 31

Food

Column: YMC-Triart C18 (5 µm, 12 nm)Dimension: 250 x 4.6 mm IDPart No.: TA12S05-2546WTEluent: A) water / formic acid (90/10) B) acetonitrile / methanol / water / formic acid (22.5/22.5/40/10)Gradient: 20-28% B (0-30 min), 28-70% B (30-40 min), 100% B (40-45 min)Flow rate: 1.0 ml/minTemperature: 25 °CDetection: UV/VIS at 535 nmSample: commercial bilberry powder (1.25 mg/ml)

Analysis of anthocyanins and anthocyanidins* 1. Delphinidin-3-O-galactoside 2. Delphinidin-3-O-glucoside 3. Cyanidin-3-O-galactoside 4. Delphinidin-3-O-arabinoside 5. Cyanidin-3-O-glucoside 6. Petunidin-3-O-galactoside 7. Cyanidin-3-O-arabinoside 8. Petunidin-3-O-glucoside 9. Peonidin-3-O-galactoside 10. Petunidin-3-O-arabinoside 11. Delphinidin 12. Peonidin-3-O-glucoside 13. Malvidin-3-O-galactoside 14. Peonidin-3-O-arabinoside 15. Malvidin-3-O-glucoside 16. Malvidin-3-O-arabinoside 17. Cyanidin 18. Petunidin 19. Peonidin 20. Malvidin

Anthocyanins: Indicated in blackAnthocyanidins: Indicated in red

Column: YMC-Triart C18 (5 µm, 12 nm)Dimension: 150 x 2.0 mm IDPart No.: TA12S05-1502WTEluent: A) water / TFA (100/0.1) B) acetonitrile / TFA (100/0.1) 20-45% B (0-25 min)Flow rate: 0.2 ml/minTemperature: 37 °CDetection: UV at 220 nmInjection: 2 ml (0.075 ~ 0.25 mg/ml)

1. Oxytocin2. Met-Enkephalin3. Leu-Enkephalin4. Neurotensin5. γ-Endorphin6. β-Endorphin

(MW 1,007)(MW 574)(MW 556)(MW 1,673)(MW 1,859)(MW 3,465)

Peptides (MW 556 - 3,465)*

Peptides

32 YMC-Triart

HILIC

Great stability and reproducibility at high pH*

Stability in high pH (pH 11, 50 °C)**

50 hr

100 hr

150 hr

200 hr

min4 5 6 7

initial0%

20%

40%

60%

80%

100%

0 50 100 150 200

Triart Diol-HILIC

Silica based Diol

Cytosine

YMC-Triart Diol-HILC offers highly reproducible separations even at high pH and high temperature. The lifetime of YMC-Triart Diol-HILIC is much longer than that of conventional silica-based Diol columns.

1. Uracil2. Uridine3. Adenosine4. Cytosine5. Cytidine6. Guanosine

1

2 3

4

5

6

min0 2 4 6 8 10 12 14

mAU

0

5

10

15

20

25

V120501A2

1

2

NH

NH NH

NH

NH2 ClH・

NH

NH NH

NH

NH2 ClH・

NH2 NH NCH3

CH3

NHNH

ClH・NH2 NH NCH3

CH3

NHNH

ClH・

Phenformin hydrochloride

Metformin hydrochloride

U120529D

1 2

0 1.00.2 0.4 0.6 0.8 1.2 min

0

10

20

30

40

mAU

Nucleosides and bases* Diabetes drugs*

Column: YMC-Triart Diol-HILIC (5 µm, 12 nm) 150 x 3.0 mm IDPart No.: TDH12S05-1503WTEluent: 100 mM CH3COONH4 / acetonitrile (10/90)Flow rate: 0.425 ml/minTemperature: 30 °CDetection: UV at 254 nmInjection: 2 µl (5 ~ 10 µg/ml)

Column: YMC-Triart Diol-HILIC (1.9 µm, 12 nm) 50 x 2.0 mm IDPart No.: TDH12SP9-0502PTEluent: 100 mM HCOOH-HCOONH4 (pH 3.7) / acetonitrile (10/90)Flow rate: 0.8 ml/minTemperature: 25 °CDetection: UV at 235 nmInjection: 2 µl (10 µg/ml)

Column: 5 µm, 150 x 4.6 mm IDPart No.: TDH12S05-1546WTEluent: acetonitrile / water / NH3 (90/10/0.1) pH 11.3

Flow rate: 1.0 ml/minTemperature: 50 °CSample: Cytosine

Rat

e on

N [%

]

Time [h]** pH < 10 is recommended for regular use


YMC-Triart 33

HILIC

Polar and hydrophilic compounds*

0

100

200

mV

0.0 2.5 5.0 min

12

Erythorbic acid(D-Isoascorbic acid)

L-Ascorbic acid

F120425A

0

100

200

mV

0.0 2.5 5.0 7.5 10.0 min

1 2

B120427L

O

OH OH

OOH

OH

H O

OH OH

OOH

OH

H1 2

20 mM KH2PO4-H3PO4 (pH 2.5)

RP modeYMC-Triart C18

100 mM HCOOH-HCOONH4 (pH 3.3) /acetonitrile (10/90)

HILIC modeYMC-Triart Diol-HILIC

Column: 5 µm, 150 x 3.0 mm IDFlow rate: 0.425 ml/minTemperature: 40 °CDetection: UV at 254 nmInjection: 4 µl (0.05 mg/ml)

YMC-Triart C18 (RP) shows very weak retention and poor resolution of L-ascorbic acid and its stereoisomer (erythorbic acid) even if 100% aqueous mobile phase is used. However, YMC-Triart Diol-HILIC shows strong retention and good resolution of these compounds with mobile phase containing 90% organic solvent.

34 YMC-Triart

QC Data

Uniform spherical particles

YMC-Triart X-Bridge HILIC

The new uniform spherical particle support is used for YMC-Triart C18 and C8. The particle is pro-duced using micro-reactor technology for the granulation process. This results in reduction of the back-pressure and leads to more reproducibility in surface modification.

Low column back-pressure*

Column: YMC-Triart C18, 5 µm, 150 x 4.6 mm IDPart No.: TA12S05-1546WTEluent: acetonitrile / water or methanol / waterFlow rate: 1.0 ml/minTemperature: 25 °C

The revolutionary production technique, adapted from micro-reactor flow technology, produces a multi-layered silica/organic hybrid stationary phase, with outstanding narrow pore size and particle size distributions which result in low back pressures.

YMC-Triart is designed for use under a wide range of conditions. Elution with higher viscosity methanol (compared with acetonitrile), YMC-Triart generates lower pressure (approx 30% lower than with conventional phases).

YMC-Triart C18, methanol/H2O

YMC-Triart C18, acetonitrile/H2O

YMC-Pack Pro C18, methanol/H2O

YMC-Pack Pro C18, acetonitrile/H2O

YMC-Triart: Improved quality of particles


YMC-Triart 35

QC Data

Batch-to-Batch reproducibility

Lot. E

Lot. D

Lot. C

Lot. B

Lot. Amin17.515.012.510.07.55.02.50.0

13

2

J091201Y

Lot. E

Lot. D

Lot. C

Lot. B

Lot. Amin14.012.010.08.06.04.02.00.0

1 2

J091201X

Excellent reproducibility of YMC-Triart phases is available even for the analysis for basic and coordination compounds which normally exhibit tailing and adsorption effects.

Basic compounds* Coordination compounds*

The reproducibility of packed columns is shown below in terms of theoretical plate number (N) and tailing factor (Tf). YMC-Triart packed columns exhibit a very narrow range of variation.

Column: YMC-Triart C18, 5 µm, 150 x 3.0 mm IDPart No.: TA12S05-1503WTEluent: 20 mM KH2PO4 (pH 6.9) / acetonitrile (65/35)Flow rate: 0.425 ml/minTemperature: 40 °CDetection: UV at 235 nm

Column: YMC-Triart C18, 5 µm, 150 x 3.0 mm IDPart No.: TA12S05-1503WTEluent: acetonitrile / 0.1% H3PO4 (40/60)Flow rate: 0.425 ml/minTemperature: 40 °CDetection: UV at 254 nm

20000

15000

10000

5000

0

2.5

2.0

1.5

1.0

0.510 20 30 40 50

Theoretical plate number

Theo

retic

al p

late

num

ber

Tailing factor

Column number

Tailing factor

Column: YMC-Triart C18, 5 µm, 150 x 4.6 mm IDPart No.: TA12S05-1546WTEluent: acetonitrile / water (40/60)

Flow rate: 1.0 ml/minTemperature: ambientSample: butyl benzoate

1. Chlorpheniramine (basic)2. Dextromethorphan (basic)3. Propyl p-hydroxybenzoate

1. Hinokitiol (coordination)2. Methyl benzoate

36 YMC-Triart

QC DataNarrow pore distribution

This figure shows the pore size distributions of some competitive materials. Comparing the pore size distributions of some competitive materials shows that YMC-Triart has a narrower distribution which results in sharper peak shapes.

In order to prevent peak errors, there is the limit to the injection volume when the sample is injected in high elution solvents (such as 100% acetonitrile). Compared with traditional columns, more than double the injection volume can injected into YMC-Triart columns as a result of the extremely narrow particle size distribution.

Improved loadability

Column: 5 µm, 50 x 2.0 or 2.1 mm IDEluent: A) water / formic acid (100/0.1) B) acetonitrile / formic acid (100/0.1) 5% B (0-0.5 min), 5-100% B (0.5-2.5 min)Flow rate: 0.4 ml/minTemperature: 40 °CDetection: UV at 275 nm

1

2

1 2

min5.20.25.1

Hybrid-silica based C18

Silica-based C18

YMC-Triart C181 2

Peak anomaly at 1.5 µL

Conventional silica-based particle

35302520Pore size (nm)

151050

4500

4000

3500

3000

2500

2000

1500

1000

500

0

Dgol

d/pV

d

X-Bridge HILIC

YMC-Triart C18 particle

YMC-Triart C18

solvent for sampleacetonitrile

injection volume1.0 µl1.5 µl2.0 µl3.0 µl

Influence of injection volume on peak shapes

.1OCH2CHCH2NHCH(CH3)2

Propranolol (50 µg/ml)

OH

.2H3CO

H3CO

CH3

CH3

(CH2)3NCH2CH2

CH(CH3)2

OCH3

OCH3

Verapamil (50 µg/ml)

CN

C


YMC-Triart 37

Multi-stage endcapping

Before

EC 1STEP

2STEP

3STEP

After bonding the alkyl chain, there are highly reactive and less reactive silanols on the surface. In traditional bonding processes, these are reacted with a single capping-compound in one step. However, the highly reactive silanols can be hydrolysed easily which contributes to the poor stability. The less reactive silanols are hard to endcap which results in poor resolution due to peak tailing.

YMC-Triart C18 and C8 use a new innovation in end capping called “multistage end-capping” for its surface modification process. By using a number of compounds with different reactivities in successive steps, all silanols can be capped to the maximum extent.

Column: 5 µm, 150 x 3.0 mm IDPart No.: TA12S05-1503WTEluent: 20 mM KH2PO4-K2HPO4 (pH 6.9) / acetonitrile (65/35)Flow rate: 0.425 ml/minTemperature: 40 °CDetection: UV at 235 nm

Ingredients in cough/cold medication:

1. Chlorpheniramine2. Dextromethorphan3. Propyl p-hydroxybenzoate

Hydroxyl group

End-capping (EC) group

The chromatographic result of a ”good“ end-capping is demonstrated:

YMC-Triart C18End-cappingby traditional method

Peak tailing for basic compounds

QC Data

38 YMC-Triart

QC Data

Column: 5 µm, 150 x 3.0 or 150 x 4.6 mm IDEluent: methanol / 0.1% H3PO4 (5/95)Flow rate: 0.425 ml/min for 3.0 mm ID 1.0 ml/min for 4.6 mm IDTemperature: 37 °CDetection: UV at 210 nm

YMC-Triart phases is synthesised using methodology adapted from micro-reactor technology. This technique ensures a reduction of impurities that contribute to peak tailing during the analysis of some types acidic compounds.

Acidic compounds*

Basic compounds*

Column: 5 µm, 150 x 3.0 or 150 x 4.6 mm IDEluent: 20 mM KH2PO4-K2HPO4 (pH 6.9) / acetonitrile (65/35)Flow rate: 0.425 ml/min for 3.0 mm ID 1.0 ml/min for 4.6 mm IDTemperature: 40 °CDetection: UV at 235 nm

The innovative surface modification technology results in excellent peak shapes even for basic compounds that often exhibit peak tailing with conventional silica- and hybrid silica-based reversed phase columns.

Ingredients in a cough/cold medication

1. Chlorpheniramine2. Dextromethorphan3. Propyl p-hydroxybenzoate

Coordinating compounds*

Column: 5 µm, 150 x 3.0 or 150 x 4.6 mm IDEluent: acetonitrile / 0.1% H3PO4 (40/60)Flow rate: 0.425 ml/min for 3.0 mm ID 1.0 ml/min for 4.6 mm IDTemperature: 40 °CDetection: UV at 254 nm

YMC-Triart phases have an extremely low level of metal impurities, much lower than conventional products, ensuring excellent peak shape for coordination compounds.

Hinokitiol

1. Hinokitiol2. Methyl benzoate

YMC-Triart C18150 x 3.0 mm ID

silica based C18Inertsil

150 x 4.6 mm ID

hybrid-silica based C18XBridge

150 x 4.6 mm ID


hybrid-silica based C18Gemini NX

150 x 4.6 mm ID

hybrid-silica based C18XBridge

150 x 4.6 mm ID


Organic acid

1. Formic acid2. Acetic acid3. Propionic acid

excellent peak sh

ape

low metal impurities


YMC-Triart 39

YMC-Triart C1850 X 2.0 mmI.D.

methanol acetonitrile

Silica based C18Brand C150 X 2.1 mmI.D.

Silica based C18Brand D550 X 2.0 mmI.D.

Silica based C18Brand I650 X 2.1 mmI.D.

methanol acetonitrile

Tf 0.97

Tf 1.21

Tf 0.93

Tf 1.03

Tf 1.14

Tf 1.11

Tf 1.56

Tf 2.83

Tf 3.87

Tf 1.88

Tf 1.03

Tf 1.13

Tf 1.05

Tf 1.13

Tf 1.22

Tf 1.16

Tf 1.43

Tf 2.21

Tf 1.88

Tf 1.78

nim0.80.80.80.70.60.210.110.01 7.06.09.0 10.0 minminmin

70G137090F70F137090F70I137090F70H137090F

70I227090F70H227090F 70G227090F70F227090F

F090714G07F090714F07F090717I07F090717H07

F090803G07F090727F07F090727I07F090727H07

F090817G07F090818F07F090818I07F090818H07

10 mM phosphate buffer (pH 6.7)/organic solvent 10 mM CH3COONH4/organic solvent

Comparison of clemastine analysis*

Column: 5 µm, 50 x 2.0 or 50 x 2.1 mm IDEluent: A) 10 mM KH2PO4-K2HPO4 (pH 6.7) or 10 mM CH3COONH4 B) methanol or acetonitrile 5-90% B (0-10 min), 90% B (10-15 min)Flow rate: 0.2 ml/minTemperature: 25 °CDetection: UV at 230 nm

Clemastine is a well-known basic compound which readily exhibits peak tailing with conventional ODS columns. YMC-Triart C18 provides sharp separations with many different buffer/solvent compositions.

Silica based C18Inertsil ODS-3V50 x 2.1 mm ID


Silica based C18Capcell Pak MG-II50 x 2.0 mm ID

Silica based C18Sunfire C1850 x 2.1 mm ID

Hybrid-silica based C18X-Bridge50 x 2.1 mm ID

QC Data

40 YMC-Triart

Ordering Information

Phase Column ID(mm) Column length (mm) Guard cartridges*

with 10 mm length

50 75 100 150 250 (pack of 5)

C182.0 TA12S05-0502WT TA12S05-L502WT TA12S05-1002WT TA12S05-1502WT — TA12S05-01Q1GC3.0 TA12S05-0503WT TA12S05-L503WT TA12S05-1003WT TA12S05-1503WT — TA12S05-0103GC4.6 TA12S05-0546WT TA12S05-L546WT TA12S05-1046WT TA12S05-1546WT TA12S05-2546WT TA12S05-0104GC

C82.0 TO12S05-0502WT TO12S05-L502WT TO12S05-1002WT TO12S05-1502WT — TO12S05-01Q1GC3.0 TO12S05-0503WT TO12S05-L503WT TO12S05-1003WT TO12S05-1503WT — TO12S05-0103GC4.6 TO12S05-0546WT TO12S05-L546WT TO12S05-1046WT TO12S05-1546WT TO12S05-2546WT TO12S05-0104GC

HILIC2.0 TDH12S05-0502WT TDH12S05-L502WT TDH12S05-1002WT TDH12S05-1502WT — TDH12S05-01Q1GC3.0 TDH12S05-0503WT TDH12S05-L503WT TDH12S05-1003WT TDH12S05-1503WT — TDH12S05-0103GC4.6 TDH12S05-0546WT TDH12S05-L546WT TDH12S05-1046WT TDH12S05-1546WT TDH12S05-2546WT TDH12S05-0104GC

PhaseColumn

ID(mm)

Column length (mm)Guard cartridges*with 5 mm length

20 30 50 75 100 150 (pack of 3)

C182.0 TA12SP9-0202PT TA12SP9-0302PT TA12SP9-0502PT TA12SP9-L502PT TA12SP9-1002PT TA12SP9-1502PT TA12SP9-E5Q1CC**3.0 — TA12SP9-0303PT TA12SP9-0503PT TA12SP9-L503PT TA12SP9-1003PT TA12SP9-1503PT TA12SP9-E503CC

C82.0 TO12SP9-0202PT TO12SP9-0302PT TO12SP9-0502PT TO12SP9-L502PT TO12SP9-1002PT TO12SP9-1502PT TO12SP9-E5Q1CC**3.0 — TO12SP9-0303PT TO12SP9-0503PT TO12SP9-L503PT TO12SP9-1003PT TO12SP9-1503PT TO12SP9-E503CC

HILIC2.0 TDH12SP9-0202PT TDH12SP9-0302PT TDH12SP9-0502PT TDH12SP9-L502PT TDH12SP9-1002PT TDH12SP9-1502PT TDH12SP9-E5Q1CC**3.0 — TDH12SP9-0303PT TDH12SP9-0503PT TDH12SP9-L503PT TDH12SP9-1003PT TDH12SP9-1503PT TDH12SP9-E503CC

YMC-Triart 1.9 µm UHPLC columns

YMC-Triart 3 µm analytical columns

YMC-Triart 5 µm analytical columns

PhaseColumn ID

(mm)Column length (mm)

Guard cartridges*with 10 mm length

50 75 100 150 250 (pack of 5)

C182.0 TA12S03-0502WT TA12S03-L502WT TA12S03-1002WT TA12S03-1502WT — TA12S03-01Q1GC3.0 TA12S03-0503WT TA12S03-L503WT TA12S03-1003WT TA12S03-1503WT — TA12S03-0103GC4.6 TA12S03-0546WT TA12S03-L546WT TA12S03-1046WT TA12S03-1546WT TA12S03-2546WT TA12S03-0104GC

C82.0 TO12S03-0502WT TO12S03-L502WT TO12S03-1002WT TO12S03-1502WT — TO12S03-01Q1GC3.0 TO12S03-0503WT TO12S03-L503WT TO12S03-1003WT TO12S03-1503WT — TO12S03-0103GC4.6 TO12S03-0546WT TO12S03-L546WT TO12S03-1046WT TO12S03-1546WT TO12S03-2546WT TO12S03-0104GC

HILIC2.0 TDH12S03-0502WT TDH12S03-L502WT TDH12S03-1002WT TDH12S03-1502WT — TDH12S03-01Q1GC3.0 TDH12S03-0503WT TDH12S03-L503WT TDH12S03-1003WT TDH12S03-1503WT — TDH12S03-0103GC4.6 TDH12S03-0546WT TDH12S03-L546WT TDH12S03-1046WT TDH12S03-1546WT TDH12S03-2546WT TDH12S03-0104GC

*Guard cartridge holder required, part no. XPGCH-Q1

*Guard cartridge holder required, part no. XPGCH-Q1

For other dimensions please refer to page 247

*Guard cartridge holder required, part no. XPCHUHP**Guard cartridge: 2.1 mm ID

42 YMC-Actus

YMC-Actus 43


Contents

•Stability ..................................................................... 44

•AxialCompressionTechnology ................................. 45

•Application ...........................................................46-47

•OrderingInformation ............................................48-49

YMC-Actus

IntroductionFast semi-preparative chromatography

Semi-preparativechromatographyisthelinkbetweenanalyticalHPLCandpreparativeLC.Eventhoughthechroma-tographicsystemsusedforsemi-preparativeLCarenotaslargeaspreparativeLCsystems,theobjectivesremainthesame:

• Purificationandisolationofmaximumsamplequantity

• Savingsintimeandcosts.

With YMC-Actus, time is on your side!

44 YMC-Actus

Stability

Rapidpressurechangesunderhigh-speedgradientconditionsmayleadtocolumndegradationandlossofcolumnperformance.

WithYMC-Actus, a newhardware andpacking technology has been applied to these semi-preparativecolumnstoprovideauniformpackingdensitywhichresultsinalongerlifetimethanconventionalsemi-preparativecolumns.

Inordertodeterminethequalityofthepacking,thecolumnperformancehasbeenevaluatedevery100runswithasequentialhigh-throughputgradient.

YMC-Actuscolumnsofferoutstandingefficiencywithoutcompromisingresolution.Furthermore,YMC-Actuscolumnsprovidereliableresults,evenafterexposuretosevere,rapidgradientcondi-tionsandmultipleinjections.

Sequential gradient testColum: 50x20mmID,5µmEluent: A)waterB)methanolGradient: 5%B(0-0.5min),5-95%B(0.5-3.1min), 95%B(3.1-3.6min),5%B(3.6-4.0min)Flowrate: 50ml/minTemp.: ambient

Column performance testColumn: 50x20mmID,5µmEluent: methanol/water(70/30)Flowrate: 10ml/minSample: Naphthalene

Increase column lifetime =Decrease of cost

Conventional semi-preparative column

YMC-Actus

YMC-Actus 45


YMC-Actus

Conventional Column Packing

withoutcompression

FlowVoidandcrack

Degradationofperformance

after300runs

initial

Axial Compression Technology for Ultimate Separation

YMC-Actusseriescolumnsaresemi-preparativeHPLCcolumnsthathaveexcellentcolumnstabilityandefficiencyasaresultofapplyingaxialcompressiontechnology.YMC-ActusseriescolumnsshowhighstabilityunderhighflowrateorsteepgradientconditionswhicharedesirableformilligramscalepreparativeHPLCofvariouscompounds.

10%increasein bed density

Compressing Flow

after700runs

initial

Long LastingColumns!

Testconditions(high-speed and high-pressure)Column: 5µm,50x20mmIDEluent: A)waterB)methanolGradient: 5%-95%BFlowrate: 50ml/minPressure: ~17MPa

UniformlyhighdensitypackingisnecessaryforhighlyefficientandstableHPLCcolumns.

DAC(DynamicAxialCompression)columnsarewidelyusedforpreparativeseparationinpilotorproductionscale.Thisallowsuniformlyhighdensitypackingandpreventsformationofvoids.

YMC-ActusseriescolumnshavebeendevelopedbyapplyingthisAxialCompressionTechnologytosemi-prepcolumnproduction.ThecolumnbediscompressedappropriatelywhenattachingtheinletendassemblyofthenewlydesignedYMC-Actushardware.Itprovidesincreasedbeddensity(10%higherthanconventionalcolumns)andbeduniformity.

46 YMC-Actus

YMC-Actus

Available for high-throughput purification: Injection in DMSO

Asshowninoverlaychromatograms,YMC-Actuscolumnsprovideoutstandingstabilityandreproducibilityinthesepa-rationofpharmaceuticalsdissolvedin100%DMSO,evenafter1,000injectionsunderthetestconditions.ThismakesYMC-Actuscolumnsidealforhigh-throughputpurificationindrugdiscovery.

Column: YMC-ActusProC18RS(5µm,8nm),50x20mmIDEluent: A)water/TFA(100/0.1) B)acetonitrile/TFA(100/0.1)Gradient: 10-90%B(0-3min),90%B(3-5min),10%B(5-10min)Flowrate: 20ml/minTemperature: ambientDetection: UVat270nmInjection: 100µl(0.2-2mg/mlinDMSO)

1.Ethenzamide2.Clemastinefumarate3.Ibuprofen

Excellent stability and efficiency under fast gradient conditions at high flow rate

Glycyrrhizic acid in herb medicine*

Eluent: A)water/aceticacid(99/1) B)methanol/aceticacid(99/1) 20%B(0-2min),20-45%B(2-7min),45%B(7-10min)Temperature: ambientDetection: UVat260nmSample: water/methanol/aceticacidextractofcommercially availableherbmedicine(0.1g/ml)

Analysis Purification

YMC-Actus 47


YMC Actus

Outstanding separation of highly polar compounds

Crude synthetic 30mer oligonucleotide*

purity>99%

Analysis Purification

1.0ml/min,5µlinjection 19ml/min,100µlinjection

30mer30mer

Impurities

Recovery 56%

50x4.6mmID,5µm 50x20mmID,5µm

Hydrosphere C18 YMC-Actus Hydrosphere C18

Excellent separation of compounds with similar structure

Analysis 50x4.6mmID,5µm2.0ml/min,20µlinjection

Capsaicinoids in red pepper*

DihydrocapsaicinCapsaicin Nordihydrocapsaicin

Purification

Purity >99.5%

YMC-Actus Pro C18 RS, 50 x 20 mm ID 5 µm

40ml/min,400µlinjection

PhenomenexLunaC18 WatersXBridgeC18

YMC-Pack Pro C18 RS

Eluent: A)methanol/water/TFA(50/50/0.1) B)methanol/TFA(100/0.1) 0-30%B(0-5min),30%B(5-10min)Temperature: 25°Cfor50x4.6mmID ambientfor50x20mmIDDetection: UVat280nmSample: methanolextractofacommercialredpepper(1g/3ml)

Eluent: A)10mMDBA-aceticacid(pH6.0)/methanol(60/40) B)10mMDBA-aceticacid(pH6.0)/methanol(20/80) 10-35%B(0-30min)Temperature: ambientDetection: UVat269nmSample: syntheticoligonucleotide(100µM)

48 YMC-Actus


Packing materialParticle size

[µm]Pore size [nm]

Column sizelength x ID [mm]

Product Code

Triart C18 S-5 12

50 x 20 TA12S05-0520WX 100 x 20 TA12S05-1020WX 150 x 20 TA12S05-1520WX 250 x 20 TA12S05-2520WX 50 x 30 TA12S05-0530WX 75 x 30 TA12S05-L530WX 100 x 30 TA12S05-1030WX 150 x 30 TA12S05-1530WX 250 x 30 TA12S05-2530WX 100 x 50 TA12S05-1053DX 150 x 50 TA12S05-1553DX 250 x 50 TA12S05-2553DX

Pro C18 S-5 12

50 x 20 AS12S05-0520WX 100 x 20 AS12S05-1020WX 150 x 20 AS12S05-1520WX 250 x 20 AS12S05-2520WX 50 x 30 AS12S05-0530WX 75 x 30 AS12S05-L530WX 100 x 30 AS12S05-1030WX 150 x 30 AS12S05-1530WX 250 x 30 AS12S05-2530WX 100 x 50 AS12S05-1053DX 150 x 50 AS12S05-1553DX 250 x 50 AS12S05-2553DX

Hydrosphere C18 S-5 12

50 x 20 HS12S05-0520WX 100 x 20 HS12S05-1020WX 150 x 20 HS12S05-1520WX 250 x 20 HS12S05-2520WX 50 x 30 HS12S05-0530WX 75 x 30 HS12S05-L530WX 100 x 30 HS12S05-1030WX 150 x 30 HS12S05-1530WX 250 x 30 HS12S05-2530WX 100 x 50 HS12S05-1053DX 150 x 50 HS12S05-1553DX 250 x 50 HS12S05-2553DX

Pro C18 RS S-5 8

50 x 20 RS08S05-0520WX 100 x 20 RS08S05-1020WX 150 x 20 RS08S05-1520WX 250 x 20 RS08S05-2520WX 50 x 30 RS08S05-0530WX 75 x 30 RS08S05-L530WX 100 x 30 RS08S05-1030WX 150 x 30 RS08S05-1530WX 250 x 30 RS08S05-2530WX 100 x 50 RS08S05-1053DX 150 x 50 RS08S05-1553DX 250 x 50 RS08S05-2553DX

Triart C8 S-5 12

50 x 20 TO12S05-0520WX 100 x 20 TO12S05-1020WX 150 x 20 TO12S05-1520WX 250 x 20 TO12S05-2520WX 50 x 30 TO12S05-0530WX 75 x 30 TO12S05-L530WX 100 x 30 TO12S05-1030WX 150 x 30 TO12S05-1530WX 250 x 30 TO12S05-2530WX 100 x 50 TO12S05-1053DX 150 x 50 TO12S05-1553DX 250 x 50 TO12S05-2553DX

Pro C8 S-5 12

50 x 20 OS12S05-0520WX 100 x 20 OS12S05-1020WX 150 x 20 OS12S05-1520WX 250 x 20 OS12S05-2520WX 50 x 30 OS12S05-0530WX 75 x 30 OS12S05-L530WX 100 x 30 OS12S05-1030WX 150 x 30 OS12S05-1530WX 250 x 30 OS12S05-2530WX 100 x 50 OS12S05-1053DX 150 x 50 OS12S05-1553DX 250 x 50 OS12S05-2553DX

High durability semi-preparative columns

YMC-Actus 49



Guard cartridges

Guard cartridges holder:20mmguardcolumnID:XPCHSPW2;30mmguardcolumnID:XPCHSPW3


[µm]Pore size [nm]


Product Code

Triart C18 S-5 12 10 x 20 TA12S05-0120CC10 x 30 TA12S05-0130CC

Pro C18 S-5 12 10 x 20 AS12S05-0120CC10 x 30 AS12S05-0130CC

Hydrosphere C18 S-5 12 10 x 20 HS12S05-0120CC10 x 30 HS12S05-0130CC

Pro C18 RS S-5 8 10 x 20 RS08S05-0120CC10 x 30 RS08S05-0130CC

Pro C8 S-5 12 10 x 20 OS12S05-0120CC10 x 30 OS12S05-0130CC

ODS-A S-5 12 10 x 20 AA12S05-0120CC10 x 30 AA12S05-0130CC

ODS-AQ S-5 12 10 x 20 AQ12S05-0120CC10 x 30 AQ12S05-0130CC

Triart Prep C18-SS-10 12 10 x 20 TAS12S11-0120CC

10 x 30 TAS12S11-0130CC

S-15 12 10 x 20 TAS12S16-0120CC10 x 30 TAS12S16-0130CC

Triart Prep C8-S S-10 20 10 x 20 TOS20S11-0120CC10 x 30 TOS20S11-0130CC

Omega S-10 — 10 x 20 OMG99S11-0120CC10 x 30 OMG99S11-0130CC

YMCbasic S-5 — 10 x 20 BA99S05-0120CC10 x 30 BA99S05-0130CC


[µm]Pore size [nm]


Product Code

ODS-A S-5 12

50 x 20 AA12S05-0520WX 100 x 20 AA12S05-1020WX 150 x 20 AA12S05-1520WX 250 x 20 AA12S05-2520WX 50 x 30 AA12S05-0530WX 75 x 30 AA12S05-L530WX 100 x 30 AA12S05-1030WX 150 x 30 AA12S05-1530WX 250 x 30 AA12S05-2530WX 100 x 50 AA12S05-1053DX 150 x 50 AA12S05-1553DX 250 x 50 AA12S05-2553DX

ODS-AQ S-5 12

50 x 20 AQ12S05-0520WX 100 x 20 AQ12S05-1020WX 150 x 20 AQ12S05-1520WX 250 x 20 AQ12S05-2520WX 50 x 30 AQ12S05-0530WX 75 x 30 AQ12S05-L530WX 100 x 30 AQ12S05-1030WX 150 x 30 AQ12S05-1530WX 250 x 30 AQ12S05-2530WX 100 x 50 AQ12S05-1053DX 150 x 50 AQ12S05-1553DX 250 x 50 AQ12S05-2553DX

YMCbasic S-5 —

50 x 20 BA99S05-0520WX 100 x 20 BA99S05-1020WX 150 x 20 BA99S05-1520WX 250 x 20 BA99S05-2520WX 50 x 30 BA99S05-0530WX 75 x 30 BA99S05-L530WX 100 x 30 BA99S05-1030WX 150 x 30 BA99S05-1530WX 250 x 30 BA99S05-2530WX 100 x 50 BA99S05-1053DX 150 x 50 BA99S05-1553DX 250 x 50 BA99S05-2553DX

High durability semi-preparative columns

50 YMC ProFamily

YMC ProFamily 51


Contents

•YMC-UltraHTProC18andHydrosphereC18 ............. 52-59

•General .......................................................................... 60-67 •YMC-PackPro C18 ...................................................... 68-69

•YMC-PackPro C8 ........................................................ 70-71

•YMC-PackPro C4 ........................................................ 72-73

•YMC-PackPro C18 RS ................................................. 74-77

•HydrosphereC18 .......................................................... 78-79

•OrderingInformation ..................................................... 80-81

HPLC Columns for Ultra Fast LC

Nowadays,especiallyinthepharmaceuticalindustry,theneedforUltraFastLCandRapidResolutionisstillgrowingduetothecontinuousdemandforhighthroughputanalysisinresearch&developmentandqualitycontrol.

TosatisfythedemandfortheseUltraFastLCmethods,YMCintroducedYMC-UltraHTcolumns,whichideallymatchthe latest instrumentation technology for“extra”highpressureapplicatione.g.withAgilent1200™seriesorWatersAcquityUPLC™.

Asacolumnandbulkmediasupplierwithmanyyearsofpracticalchromatographicexperience,YMCfoundunacceptablethattheuseofnovelseparationmediaisoftenrestrictedtodedicatedequipmentandnotapplicabletothelargeinstalledbaseof“conventional”HPLCsystemswithastandardpressureratingof5800psi(400bar,40MPa).Forthisreason,specificationsforYMC-UltraHTcolumnshavebeendesignedtoprovidepowerfulchromatographicimprovements,intermsofvelocityandresolution,evenwithconventionaloperatingconditions.SinceYMC-UltraHTcolumnsprovidesasubstantiallylowerpressuredropthanmostcompetitive2µmorsub-2µmmedia,highflowratescanbeachievedwithoutgeneratingexcessivebackpressureandwithouttheneedforspecialisedequipment(seepage54fordetails).

Foreffectivehighthroughputseparations,YMCofferawiderangeofhighperformanceHPLCcolumnswhichallowUltraFastanalyticalHPLCwithconventionalequipment.Duetothedown-scalabilityofthemajorityofYMC’sstationaryphases,thetimeneededforasingleanalysiscanbereducedtolessthan60seconds,dependingonthesampleconditions.

YMC ProFamily

OneofthemainchallengesinRP-HPLCisthequantitationofionisablecompoundsincludingdrugs,degradationproducts,etc.Forthispurposesymmetrical,sharppeaksarerequiredtoprovidehighestresolutionandreliableintegration.ThestationaryphasesoftheYMCProFamilyfullfilthesedemandsmakingthemanexcellentchoiceforthepharmaceuticalandbiotechnologyindustries.Thisproduct lineconsistsofthethreeC18-phases:YMC-PackProC18RS(withhighcarbonload[22%]),YMC-PackProC18andHydrosphereC18(“AQ-type”)togetherwiththeC8-andC4-phase:YMC-PackProC8andYMC-PackPro C4.

YMC ProFamily

Introduction

52 YMC ProFamily

Ultra Fast LC Columns- YMC Pack ProFamily chemistries, based on ultra high purity silica, provide excellent resolution for a wide range of analytes

- YMC-UltraHT LC Columns provide considerable time saving without resort to ultra high pressures

- YMC-UltraHT LC Columns achieve ultra fast separations even with conventional HPLC equipment

- fully up- and down-scalable selectivity

General

SincetheintroductionoftheProFamilyseriesofphases,YMC-PackProC18hasprovedtobeoneofthefirstchoicesforawiderangeofHPLCapplicationsinpharmaceuticalandbiotechnologicalresearchandproduction,whereefficiencyandreliabilityareamajordemand.

Inmanycases,theseparationofhighlypolarcompoundsrequireshighlyaqueousmobilephaseconditionstoachievesufficientretentiononthestationaryphase.Conventional reversedphaseselectivitiesdonotgivereproducibleresultsundertheseconditionsduetomainlycollapseoftheC18chains.Therefore,YMCdiddevelopedHydrosphereC18inorderoftoovercomethelossinretention.Now,thisoutstandingchromatographicbehaviourhasbeentransferredtoYMC-UltraHTHydrosphereC18.

YMC-PackProC18isawell-establishedC18silica-basedcolumn,whichprovidesamediumbal-anceofhydrogen-bondingcapacityandhydrophobicity,asshownbelow.Conversely,HydrosphereC18isoptimalselectivityfortheseparationofhighlypolarcompounds.

Specifications YMC-UltraHT Pro C18 YMC-UltraHT Hydrosphere C18Particle size / µm 2 2Pore size / nm 12 12Surface area / m2g-1 330 330Carbon content / % 16 12Recommended pH range 2.0 - 8.0 2.0 - 8.0

YMC ProFamily 53


Ultra Fast LC Columns

Van Deemter Plot*

Why smaller Particles?

EversinceinthebeginningofHPLC,more-demandinganalyticalproblemshaverequiredaprogres-siveimprovementinseparationefficiency.Thechallengesincludeevermorecomplexanalytesandthereductioninanalysistimestokeepupwiththeincreasingnumbersofsamples.Inadditiontoreducingthecolumndimensionsandincreasingflowrates,theimplementationofsmallparticlesisapowerfultooltoincreaseefficiency.

0.000

0.005

0.010

0.015

0.020

0.000

0.005

0.010

0.020

HE

TP [m

m/p

late

s]

YMC-Pack Pro C185 µm


Optimum flow rate for 2 µm(0.3 - 0.8 ml/min for 2 mm ID)

YMC-UltraHT Pro C182 µm

0 1 2 3 4 5 6 7 8 9 100 1 2 3 4 5 6 7 8 9 10 u [mm/sec]

0 0.2 0.4 0.6 0.8 1.0 1.1 1.3 1.5 F [ml/min]1.7 1.9

11

2.1

12

2.3

ThevanDeemterequationdescribesthe“HeightEquivalentoftheTheoreticalPlate”(HETP)asafunctionofthelinearvelocity(u)by

H = A+B/u+C*uwhereA,BandCareconstantsanduisthemobilephaselinearvelocitymeasuredinmm/sec.

TheresultingvanDeemterplotsshowthereductionofHETPwhenusingsmallerparticlesizesofYMC-PackProC18withanadditionalshiftoftheminimumvaluetohigherflowrates.

Selectivity-Chart*

0.04

0.06

0.08

0.10

0.12

0.14

0.16

0.18

2 4 6 8 10 12 14

��

Hydrogen-bonding capacity: high

Hydrophobicity: high

Hydrogen-bonding capacity: low

Hydrophobicity k’(Amylbenzene)

Hyd

rog

en-b

ond

ing

cap

acity

a(C

affe

ine

/ B

enze

ne)

0.04

0.06

0.08

0.10

0.12

0.14

0.16

0.18

2 4 6 8 10 12 14

��

��

��

Hydrophobicity: medium

Hydrogen-bonding capacity: medium

Hydrophobicity: low

Hydrogen-bonding

Hydrogen-bonding

Hyd

rog

en-b

ond

ing

cap

acity

a



YMC-UltraHT Pro C182 µm

54 YMC ProFamily

Features of Packing MaterialWhenstartingtofocusonUltraFastLCthroughtheuseofsmallparticles,veryhighbackpres-sureshavetobeconsideredandabalancesought.TheextensiveexperienceinsilicaproductionenablesYMCtoprovidesmallparticleswithanextremelynarrowparticlesizedistributionwhichresultsinlowbackpressures.

YMC’sUltraHTProC18columnsofferoutstandingefficiencyforFastLCwithoutexhibitingextremelyhighbackpressurevalueswhichcanbeobtainedwithsub-2µmparticlesfromothermanufactur-ers.ThereforeYMC’sUltraHTProC18maynotrequirededicatedHPLCequipmentforprovidingoutstandingcolumnperformances.

Column Pressure*

Columns: 50x2.0or2.1mmIDEluent: acetonitrile/water(60/40)Temperature: 25°CSample: Butylbenzoate

Lower Column

Pressure!

TheintroductionofYMC-PackProC182µmallowseasydownscalingofexistingmethodswhichuseYMC-PackProC183µmand5µm.

Reproducibility Comparison*

Column: 50x2.0or2.1mmIDEluent: acetonitrile/water(60/40)Temperature: 25°CDetection: UVat270nm

Fully Up- and

Downscalable!

0.0 1.0 2.0 3.0 4.0 5.0 6.0 min

1

2

3

4

0.0 1.0 2.0 3.0 4.0 5.0 6.0 min0.0 1.0 2.0 3.0 4.0 5.0 6.0 min

1

2

3

4

1. Uracil 2. Methylbenzoate 3. Naphthalene 4. Butylbenzoate

YMC-UltraHT Pro C18 2 μm

YMC-Pack Pro C18 3 μm

YMC-Pack Pro C18 5 μm

0

10

20

30

40

60

80

00 11 22 33 44 55 66 77 88

��

��

��

��

��

Col

umn

Pre

ssur

e (M

Pa)

u [mm/sec]

Conventional HPLCconditions

0 0.2 0.4 0.6 0.8 1.0 1.1 1.3 1.5 F [ml/min]

70

50

YMC ProFamily 55


Features of Packing MaterialThegraphbelowshowsthedependencyof“HeightEquivalentoftheTheoreticalPlate”(HETP)andthelinearvelocityinthepresenceofdifferentorganicsolvents.Whenmethanolisused,theoptimumHETPisachievedwithinadifferentrangeofvelocitycomparedtowhenacetonitrileisusedduetotheirdifferentviscosities.Thereforetheoptimumrangeofflowratechangeswiththeorganicsolvent.

Themaximumresolutionisobtainedbyoptimisingflowrate,temperature,andorganicsolventinordertoachievetheoptimumbackpressure.

Linear velocity versus HETP*

Column: YMC-UltraHTProC18,2µm,50x2.0mmIDTemperature: 25°CSample: Butylbenzoate

0.000

0.002

0.004

0.006

0.008

0.010

0.012

0.014

0 1 2 3 4 5 6 7 8 9 10

HET

P (m

m/p

late

s)

Optimum flow rate3Optimum flow rate

0.000

0.002

0.004

0.006

0.008

0.010

0.012

0.014

0 1 2 3 4 5 6 7 8 9 10

HET

P (m

m/p

late

s)

u [mm/sec]

��

60% methanol 60% acetonitrile

Linear velocity vs. HETP

with 60% CH CNwith 60% CH3OH

0 0.2 0.4 0.6 0.8 1.0 1.1 1.3 1.5 F [ml/min]1.7 1.9

Fast LC analysis with conventional LC equipment*

Column: YMC-UltraHTPro C18Eluent: acetonitrile/20mMKH2PO4 (10/90)Temperature: 37°CDetection: UVat260nm

50 x 3 mm IDSemi-microCell(2.5µl,Celllength5mm)

Flowrate:0.8ml/min

50 x 2 mm IDSemi-microCell(2.5µl,Celllength5mm)

Flowrate:0.36ml/min

StandardCell(8µl,Celllength10mm)

Flowrate:0.8ml/min

StandardCell(8µl,Celllength10mm)

Flowrate:0.36ml/min

SinceYMC-UltraHTcolumnsprovidesubstantiallylowerpressuredropthanmostcompetitive2µmorsub-2µmmedia,highflowratescanbeachievedwithoutgeneratingexcessivebackpressureandwithouttheneedforspecialisedequipment.Nevertheless,3mmIDcolumnsarelessaffectedbythediffusionvolumethan2mmIDcolumns.Therefore,itisnecessarytoreducethesystem“dead”volumeinordertoobtainoutstandingchromatographicperformanceswith2mmIDcolumns.

1. Uracil 2. Methylbenzoate 3. Naphthalene 4. Butylbenzoate

56 YMC ProFamily

Conventional LC Method*

Eluent: acetonitrile/water/formicacid(35/65/0.1)Flowrate: 1.0ml/min(1.00mm/sec)Temperature: 40°CDetection: UVat260nmPressure: 1230 psi (83 bar, 8.3 MPa)

0.0 5.0 10.0 15.0 20.0

0

50

100

0.0 5.0 10.0 15.0 20.00.0 5.0 10.0 15.0 20.0 25.025.0 min

0

50

100

mV

1 2 34 5 6

7

YMC-Pack Pro C18 5 μm 150 x 4.6 mm ID

Ultra Fast LC Method*

Eluent: acetonitrile/water/formicacid(35/65/0.1)Flowrate: 0.6ml/min(3.18mm/sec)Temperature: 40°CDetection: UVat260nmPressure: 4350 psi (292 bar, 29.2 MPa)

1. Myricetin 2. Querecetin 3. Apigenin 4. Kaempferol 5. Baicalein 6. Chrysin 7. Acacetin

1

2

3

45

6

7

3.03.0

mV

YMC-UltraHT Pro C1850 x 2.0 mm ID

DuetotheproductionprocessesusedtomanufactureYMC-PackProFamily,methodscanbeeasilydownscaledwithunchangedselectivity.

Astheexamplesshowndemonstrate,conventionalHPLCmethods can betransferredeasilytoUltraFastLCmeth-odsbychoosingYMC-UltraHTcolumns

togainefficiencyandsignificantlyreduceanalysistime.

Downscale of Methods

Peptide mapping*

Eluent: A)acetonitrile/water/TFA(10/90/0.1) B)acetonitrile/water/TFA(35/65/0.1)Flowrate: 0.2ml/minTemperature: 37°CDetection: UVat220nmSample: TrypticdigestofCytochromec(5mg/ml;37°C;24h)

0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0

�

100

200

0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0

�

100

200

0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0

�

100

200

mAU

15.0 min

0 5 10 15 20 25 30 35 40

0

40

80

45.0 min0 5 10 15 20 25 30 35 40

mAU

0

40

80

TheapplicationofHPLCtobiologicallyrelevantseparationsisanexistingandrapidlygrowingfield.YMC-UltraHTProC18providesoutstanding chromatographicperformance,which ismore thancapableofmeetingthechallengeofpeptidemapping,wherealargenumberofpeptidefragmentsaregeneratedfromenzymaticdigestion.

5 μm YMC-Pack Pro C18150 x 2.0 mm ID0%B(0-5min)0-100%B(5-40min)100%B(40-45min)

2 μm YMC-UltraHT Pro C1850 x 2.0 mm ID0%B(0-1.65min)0-100%B(1.65-13.35min)100%B(13.35-15min)

YMC ProFamily 57



Method transfer from conventional LC to Ultra Fast LC*

Columns: HydrosphereC18Eluent: A:water/aceticacid(100/3) B:acetonitrile/aceticacid(100/3)Temperature: 35°CDetection: UVat254nm

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1. Daidzin 2. Glycitin 3. Genistin 4. 6‘-O-Malonyldaidzin 5. 6‘-O-Malonylglycitin 6. 6‘-O-Acetyldaidzin

7. 6‘-O-Acetylglycitin 8. 6‘-O-Malonylgenistin 9. Daidzein10. Glycitein11. 6‘-O-Acetylgenistin12. Genistein

150x4.6mmID,5µm15-35%ACN(0-30min)1.0ml/min10µlInj.

50x4.6mmID,3µm15-35%ACN(0-10min)1.5ml/min3µlInj.

50x4.6mmID,3µm12.5-35%ACN(0-10min)1.5ml/min3µlInj.

50x2.0mmID,2µm12.5-35%ACN(0-10min)0.3ml/min0.6µlInj.

50x2.0mmID,2µm12.5-35%ACN(0-3.3min)0.9ml/min0.6µlInj.

58 YMC ProFamily


Peak performance*

2 µm YMC-Ultra HT Pro C18, 50 x 2.0 mm ID

1.7 µm Acquity UPLC BEH C18, 50 x 2.1 mm ID

1.8 µm Zorbax Eclipse XDB-C18, 50 x 2.1 mm ID

N 5 = 106,000 /mTf 5 = 1.074470 psi (300 bar, 30 MPa)

N 5 = 89,000 /mTf 5 = 3.997450 psi (500 bar, 50 MPa)

N 5 = 78,000 /mTf 5 = 2.355960 psi (400 bar, 40 MPa)

12

34

5

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

mV

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

601

23

4

5

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

mV

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

601

23

45

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

0.20.0 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min0

20

40

60

mV

Why not take the pressure out of Fast LC!

With YMC-UltraHT Pro C18 you have all the efficiency you need to develop your Fast LC methods with none of the pressure or heat some would have you believe is essential!

Peak performance*

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2 µm YMC-UltraHTHydrosphere C18,50 x 2.0 mm ID

7449 psi (500 bar, 50 MPa)

8044 psi (540 bar, 54 MPa)

11,470 psi (770 bar, 77 MPa)

2 µm YMC-UltraHT Pro C18,50 x 2.0 mm ID

1.8 µm Zorbax SB-C18,50 x 2.1 mm ID

1. Scopolamine 2. Atropine 3. Cinchonine

4. Quinine 5. Dihydroquinine

1. Daidzin 2. Glycitin 3. Genistin 4. 6‘-O-Malonyldaidzin 5. 6‘-O-Malonylglycitin 6. 6‘-O-Acetyldaidzin

7. 6‘-O-Acetylglycitin 8. 6‘-O-Malonylgenistin 9. Daidzein10. Glycitein11. 6‘-O-Acetylgenistin12. Genistein

2 µm YMC-UltraHT Pro C18, 50 x 2.0 mm ID

1.7 µm Acquity UPLC BEH C18, 50 x 2.1 mm ID

1.8 µm Zorbax Eclipse XDB-C18, 50 x 2.1 mm ID

YMC ProFamily 59



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Exctracts obtained from soy foods and dietary supplements*

1. Daidzin(D) 2. Glycitin(Gl) 3. Genistin(G) 4. 6‘-O-Malonyldaidzin(MD) 5. 6‘-O-Malonylglycitin(MGl) 6. 6‘-O-Acetyldaidzin(AD) 7. 6‘-O-Acetylglycitin(AGl) 8. 6‘-O-Malonylgenistin(MG) 9. Daidzein(De)10. Glycitein(Gle)11. 6‘-O-Acetylgenistin(AG)12. Genistein(Ge)

60 YMC ProFamily

- YMC-Pack ProFamily based on ultra high purity silica

- Hydrosphere C18 for stability in aqueous mobile phases

- every packed column supplied with: • lotcertificate • testchromatogram

YMC ProFamily

Pro C18 Pro C8 Pro C4 Pro C18 RS Hydrosphere C18Particle size / µm 2; 3; 5 3; 5 3; 5 3; 5 2; 3; 5Pore size / nm 12 12 12 8 12Surface area / m2g-1 330 330 330 510 330Carbon content / % 16 10 7 22 12pH range 2.0 - 8.0 2.0 - 7.5 2.0 - 7.5 1.0 - 10.0 2.0 - 8.0Metal content (Randomly selected lots)Al / ppm 0.3 0.2 0.6 0.3 0.7Fe / ppm 2.8 2.5 2.9 0.1 1.2Na / ppm 0.3 1.4 1.0 1.3 0.7Ti / ppm 0.1 0.1 0.1 0.1 0.1

seepages68-69

seepages70-71

seepages72-73

seepages74-77

seepages78-79

22

1-10

Lot-to-lot reproducibility of YMC-Pack Pro C18

Lot C

Lot B

Lot A

10 15 min0 5

1 2 3 4

HPLC-Test A Specification

k’(Ethylbenzene)/k’(n-Propylbenzene) 0.629-0.653

k’(4-n-Butoxybenzoicadic)/k’(n-Propylbenzene) 0.238-0.263

1.Uracil2.4-n-Butoxybenzoicacid3.Ethylbenzene4.n-Propylbenzene

HPLC Test AColumn: 150x4.6mmIDEluent: 20mMKH2PO4-H3PO4(pH3.5)/acetonitrile(40/60,v/v)Flowrate: 1.0ml/minDetection: UVat254nmTemperature: 37°C

Properties

Strictqualitycontrolisenforceddur-ingthemanufacturingoftheunderly-ing silica, bondingof the stationaryphase,endcappingandcolumnpack-ingoperationstosupplyhighperform-ance columnsof high reproduciblequalityoveralongperiodoftime.

YMC ProFamily 61


YMC ProFamilyUnderlying silica gel support

Thephysicalpropertiesofsilicagelhaveagreateffectontheselectivityandperformanceofthebondedpacking.Forthepurposeofsupplyingcolumnsofstablequality,thephysicalpropertiesofsilicagelusedforpackingsuchasparticlesize,poresize,specificsurfacearea,porevolumeandamountofmetalcontaminationhavetobestrictlycontrolled.

Particle size*

Pore size* Specific surface area*

Pore volume* Amount of metal content*

Physical properties (Pro C18, 5 μm, 12 nm) Silica Support Material (5 μm, 12 nm)

ConsecutiveLotNumber





62 YMC ProFamily

YMC ProFamilyPacking material

ExcellentreproducibilityoftheProC18isshownnotonlyintheseparationofhydrophobiccompoundsbutalsointhatofhydrophilic,basic,andacidiccompounds.

Percent Carbon Content*

Hydrophobicity* Acidic compounds*

Hydrogen bonding ability* Basic compounds*

Pro C18 5 μm, Reproducibility between batches



ConsecutiveLotNumber ConsecutiveLotNumber


YMC ProFamily 63


Individual Column TestTogiveourcustomersaninsightintothestrictcriteriawithregardtothesilicabase,thebondedfinalproductandthereproduciblechromatographicbehaviour,eachcolumnoftheProFamilyissuppliedwithalotinspectionreportandanindividualcolumntestchro-matogram.Thefirstreportillustratesthenarrowwindowforphysicalparameterssuchasparticlesizedistributionorsurfaceareaandthereproducibilityofchemicalproperties.Thetestchromatogramillustratestheefficiencyofthecolumnwithaguaranteedminimumperformanceof100,000theoreticalplatesfor150and250x4.6mmIDandanasymmetryof0.90to1.15(at10%peakheightfor5µmparticlesize).

Individual Lot Test

Indicates the efficiency of thecolumn retention characteristicsand symmetry of the test peaks

64 YMC ProFamily

YMC ProFamilyComparison of separative selectivity

Theselectivitycharacteristicsofeachcolumnareshownusinghydrophobicityandhydrogen-bondingabilityasindicators.TheProFamilyseriesofODSphasesisdesignedtomakeHydrosphereC18and ProC18RShavecontrastingseparationcharacteristics,withstandardPro C18 in between. Also, ProC8andC4havedifferentselectivityfromtheODSphases.Bychoosingonefromthese5typesofcolumns,onecaneasilyoptimisetheseparationofpolarandnon-polarcompounds.

Durability for repetitive analysis

Thelong-termstabilityofaProC18(3µm)shortcolumnusedinrepeatedanalysisisshownbelow.Thereisnochangefoundintheseparationofallcompoundsafter5000injections(8hours/dayfor5month)duringgradientanalysis.

Great durability!

0.04

0.06

0.08

0.10

0.12

0.14

0.16

0.18

2 4 6 8 10 12 14

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Hydrogen-bonding capacity: high

Hydrophobicity: high

Hydrogen-bonding capacity: low

Hydrophobicity k’(Amylbenzene)

Hyd

rog

en-b

ond

ing

cap

acity

a(C

affe

ine

/ B

enze

ne)

0.04

0.06

0.08

0.10

0.12

0.14

0.16

0.18

2 4 6 8 10 12 14

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Hydrophobicity: medium

Hydrogen-bonding capacity: medium

Hydrophobicity: low

Hydrogen-bonding

Hydrogen-bonding

Hyd

rog

en-b

ond

ing

cap

acity

a

Column: YMC-PackProC18(3µm,12nm),50x4.6mmIDEluent: A)water/formicacid/100/0.05) B)acetonitrile/formicacid(100/0.05) 10-90%B(0-3min,linear),90%B(3-5min),10%B(5-9min)Flowrate: 1.0ml/minTemperature:37°CDetection: UVat254nm

YMC ProFamily 65


YMC ProFamilyHydrophobicity and steric selectivity

ThiscomparisonshowsthedifferentpropertiesoftheProFamilymembersgivingagoodindicationontheirpotentialformethoddevelopment.

Thecompounds1.uracil(deadvolumemarker)2.n-butylbenzene3.o-terphenyl4.n-amylbenzeneand5.triphenyleneareusedtodeterminethehydrophobicity(2.and4.)andthestericselectivity(3.and.5.)ofeachProFamilymemberunderunbufferedchromatographicconditions.

YMC-Pack Pro C4

5 10 15 20 250 min

5 10 15 200 min

5 10 15 200 min

5 10 15 200 min 5 10 150 min

1

23

4

5

1

2

3

4

5

1

2

3

4

5

12

3

4

5

1

2

3

4

5

YMC-Pack Pro C18 RS

YMC-Pack Pro C18

YMC-Pack Pro C8

YMC-Pack Pro C4

Hydrosphere C18

Column: 150x4.6mmIDEluent: methanol/water(80/20)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat254nm

ThewholeProFamily covers a largeareaof hydrophobicity and steric selectivity, aspresented in this comparison,whichofferstheopportunitytoaccomplishoptimisationofchromatographicmethodsevenforcomplicatedseparationproblems.

Formoreapplicationspleaserefertoour“ApplicationDataCollections”orcontactusdirectly.

1. Uracil 2. n-Butylbenzene 3. o-Terphenyl 4. n-Amylbenzene 5. Triphenylene

YMC-Pack Pro C8

5 10 15 20 250 min

5 10 15 200 min

5 10 15 200 min

5 10 15 200 min 5 10 150 min

1

23

4

5

1

2

3

4

5

1

2

3

4

5

12

3

4

5

1

2

3

4

5

YMC-Pack Pro C18 RS

YMC-Pack Pro C18

YMC-Pack Pro C8

YMC-Pack Pro C4

Hydrosphere C18

lowerhydrophobicity

lowesthydrophobicity

Hydrosphere C18

5 10 15 20 250 min

5 10 15 200 min

5 10 15 200 min

5 10 15 200 min 5 10 150 min

1

23

4

5

1

2

3

4

5

1

2

3

4

5

12

3

4

5

1

2

3

4

5

YMC-Pack Pro C18 RS

YMC-Pack Pro C18

YMC-Pack Pro C8

YMC-Pack Pro C4

Hydrosphere C18

highpolar recognition

YMC-Pack Pro C18 RS

5 10 15 20 250 min

5 10 15 200 min

5 10 15 200 min

5 10 15 200 min 5 10 150 min

1

23

4

5

1

2

3

4

5

1

2

3

4

5

12

3

4

5

1

2

3

4

5

YMC-Pack Pro C18 RS

YMC-Pack Pro C18

YMC-Pack Pro C8

YMC-Pack Pro C4

Hydrosphere C18

highestcarbon load

5 10 15 20 250 min

5 10 15 200 min

5 10 15 200 min

5 10 15 200 min 5 10 150 min

1

23

4

5

1

2

3

4

5

1

2

3

4

5

12

3

4

5

1

2

3

4

5

YMC-Pack Pro C18 RS

YMC-Pack Pro C18

YMC-Pack Pro C8

YMC-Pack Pro C4

Hydrosphere C18

YMC-Pack Pro C18

balanced hydrogenbonding, capacityand hydrophobicity

66 YMC ProFamily

YMC ProFamilyHydrogen bonding capacity

Hydrogenbondingcapacityisevaluatedbyexaminingtherelativeretentioncoefficientasa(caffeine/benzene).AmongtheProseriesbothHydrosphereC18withlowdensityofC18,andProC4withshortalkylchainhavehighhydrogen-bondingcapacity.Benzenewithnon-polargroupsisretainedaccording tohydrophobicityof thepacking,while retentionof caffeineandphenol (hydrophiliccompounds), isgreatlyaffectedbyhydrogen-bondingcapacity,andthesepackinghavesimilarretentiontime,butshowdifferentselectivity.

Hydrogen bonding capacity*


YMC ProFamily 67


YMC ProFamilyAcidic and basic compounds

Acidic compounds*

Column: 150x4.6mmIDEluent: 20mMCH3COONa-CH3COOH(pH4.4)/acetonitrile(80/20)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat230nm

1. p-Hydroxyacetophenone(I.S.) 2. Sorbic acid 3. Dehydroaceticacid

Basic compounds*

Column: 150x4.6mmIDEluent: 20mMKH2PO4-K2HPO4(pH6.9)/methanol(30/70)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat254nm

68 YMC ProFamily

YMC-Pack Pro C18- specifically designed for pharmaceutical and biotechnical R&D

- extreme narrow specifications

- high lot-to-lot reproducibility

- high column-to-column reproducibility

- ideal for basic, acidic and polar compounds

USPL1

YMC-Pack Pro C18 SpecificationParticle size / µm 2*; 3*; 5Pore size / nm 12 Surface area / m2g-1 330Carbon content / % 16Recommended pH range 2.0 - 8.0

Properties

YMC-PackProC18isbasedonanultrapuresilicasupport,whichisusedforthewholeProFamily.Duetoaproprietaryendcappingprocessespeciallydesignedfor this typeofsilica,YMC-PackProC18isperfectlysuitablefortheseparationofacidicandbasicmolecules.Theinertnessofthesilicamakesitanexcellentchoicefortheanalysisofdrugsormetabolites,compoundsthataresusceptibletopolarinteractionswithresidualsilanolgroupsandmetalimpuritiesasdemonstratedinthefollowingcomparison.TheextremebasicsubstancesareselectedtoprovetheverygoodperformanceofYMC-PackProC18inregardtotheirseparationandthepeakperformancethatcannotbeachievedwithclassicalmaterials.

* please be referred to page 52 ff for YMC-UltraHT columns

YMC-PackPro C18YMC-PackODS-A

Basic compounds*

Tricyclic antidepressantsColumn: YMC-PackProC18(5µm,12nm)150x4.6mmIDEluent: 20mMpotassiumphosphatebuffer(pH6.9)/ methanol(25/75)Flowrate: 1.0ml/minTemperature:37°CDetection: UVat254nm

Great peak shape

LC-MS separations*

Column: YMC-PackPro C18(3µm,12nm)50x2.0mmIDEluent: 10mMCH3COONH4/methanol(45/55)Flowrate: 0.2ml/minTemperature: 37°CDetection: UVat254nm

YMC ProFamily 69


YMC-Pack Pro C18

Column care

YMCPackProC18isstabletowardshydrolysisbetweenpH2.0-8.0.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.

Fordetailedinformationpleaserefertothe“ColumnCareandUseInstructions”whichareshippedwitheachanalyticalcolumn.

1. Ethosuximide 2. Primidone 3. Phenobarbitalsodiumsalt 4. Phenytoin 5. Carbamazepine

Tetracycline antibiotics* Antiepileptics*

Cardiac glycosides*

Application

ThissmallcollectionofapplicationscanonlygiveabriefinsightintothemultipleapplicationsforPro C18.


Column: YMC-PackProC18(5µm,12nm)150x4.6mmIDPartNo.: AS12S05-1546WTEluent: acetonitrile/10mMoxalicacid(10/90)Flow: 1.0ml/minDetection: UVat280nm,0.13AUFSTemperature: 37ºCInjection: 6µl(0.2mg/ml)

Column: YMC-PackProC18(5µm,12nm)150x4.6mmIDPartNo.: AS12S05-1546WTEluent: 50mMNH4H2PO4-(NH4)2HPO4(pH6.7)/methanol(50/50)Flow: 0.7ml/minDetection: UVat215nm,0.32AUFSTemperature: 37ºCInjection: 8µl(0.035p0.7mg/ml)

Column: YMC-PackProC18(5µm,12nm)75x4.6mmIDPartNo.: AS12S05-L546WTEluent: acetonitrile/water(10/90)Flow: 1.0ml/minDetection: UVat220nm,0.26AUFSTemperature: 37ºCInjection: 5µl(0.2,0.4mg/ml)

1020min

1. Toluene 2. AmitriptylineHCl 3. Progesterone 4. Propylbenzene 5. Acenaphthene

10 20 300 min

1

2

3

5

1

2

3

4

5

4

10 200 min 100 min

1

2 3

4

5

1

2

3

5,4

Pro C18

Hydrosphere C18

Pro C8

Pro C4

12

3

4

5

10 20 30 400 min

Pro C18 RS

10 200 min

Baseline separation of compounds withdifferent polarity*

Column: YMC-PackPro C18(3µm,12nm)75x2.0mmIDPartNo.: AS12S03-L502WTEluent: methanol/water/TFA(5/95/0.5)Flow: 0.2ml/minDetection: UVat280nm,0.16AUFSTemperature: 30ºCInjection: 1µl(0.2–0.5mg/ml)

70 YMC ProFamily

YMC-Pack Pro C8- extremely broad selectivity pattern

- good alternative to C18-phases

- suitable for all types of organic molecules, especially basic pharmaceuticals

USPL7

General

WithintheProFamily,theYMC-PackProC8providesanadditional,lesshydrophobicstationaryphaseforalltypesofcompounds,butespeciallyforbasicandmetalchelatingsubstances.Formanyapplicationsregardingtheseparationofpeptides,nucleicacidsandsimilarcompoundswithLC-MSdetection,conventionalC8-stationaryphasesrequireionpairreagentsandion-suppressiontoobtainsatisfactoryseparationandlowdetectionlimits. Incontrast,ProC8withitsultrapuresilicaallowstheanalysiswithoutthesemodifiersbutstillgeneratesexcellentchromatograms.InadditiontothereducedhydrophobicityofYMC-PackProC8comparedwithYMC-PackPro C18, thedifferentstericselectivityoffersnewpossibilitiesinmethodoptimisationasdemonstratedinthefigurebelow:

1.Uracil2.Triphenylene3.o-Terphenyl

Pro C8

Pro C18

Steric selectivity YMC-Pack Pro C18 vs. YMC-Pack Pro C8*


Thereversedelutionorderoftriphe-nyleneando-terphenylfortheshorterC8chainsofYMC-PackPro C8 illus-tratesthedifferentstericrecognitionforsteric-demandingsubstances.

YMC-Pack Pro C8 SpecificationParticle size / µm 3; 5Pore size / nm 12 Surface area / m2g-1 330Carbon content / % 10Recommended pH range 2.0 - 7.5

YMC ProFamily 71


YMC-Pack Pro C8

Column care

YMC-PackProC8isstabletowardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.


1. Butylp-hydrosybenzoate(I.S.) 2. Chlorpheniraminemaleate 3. Diphenhydraminehydrochloride

Peak performance of YMC-Pack Pro C8 for basic substances*

1. Theophylline(IS) 2. Phenol 3. Benzoicacid 4. Salicylic acid

1. Uracil 2. Procainamide 3. N-Acetylprocainamide 4. Phenol

No need for ion pair reagents* Good peak performance for acidic and basic compounds*

YMC-PackProC8isamemberoftheProFamilyandasaresultgivesexcellentpeakshapesevenforbasicsubstances,duetoitslowmetalcontentandtheendcappingprocedure,whichisidenticaltothatusedforYMC-PackProC18.ThisshallbedemonstratedinthecomparisonbelowwhereYMC-PackProC8outperformscompetitivestateoftheartproducts.

Inertsil C8-3 Waters Symmetry C8


Histamine antagonistsColumn: 150x4.6mmIDEluent: 20mMKH2PO4-K2HPO4(pH6.9)/methanol(35/65)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat260nm,0.26AUFS

DisinfectantsColumn: YMC-PackProC8(5µm,12nm)150x4.6mmIDPartNo.: OS12S05-1546WTEluent: acetonitrile/20mMH3PO4(20/80)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat270nm,0.32AUFS

Basic compoundsColumn: YMC-PackProC8(5µm,12nm)150x4.6mmIDPartNo.: OS12S05-1546WTEluent: 20mMKH2PO4-K2HPO4(pH6.9)/acetonitrile(90/10)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat254nm,0.32AUFS

YMC-PackPro C8

72 YMC ProFamily

YMC-Pack Pro C4- proprietary endcapping in order to minimize the effect of residual silanols

- for polar organic molecules, especially basic pharmaceuticals and peptides

- ideal for fast chromatography

USPL26

General

Morethan80%oftheRP-HPLCanalysesareaccomplishedonoctyl-oroctadecyl-phases.Becauseof thisoverwhelmingmajority,manychromatographersneglectotherselectivities thatmightbebettersuitedtotheirseparation,suchasbutylphases.WithProC4,YMCoffersastationaryphasebasedonthewell-knownultrapuresilicaoftheProFamily.ComparedtoaC18-phasewiththesameeluent,thislesshydrophobicmaterialgivesshorterretentiontimesfornon-polarcompoundswhiletheretentiontimeofpolaranalytesarevirtuallyunaffected.ThismakestheProC4aninterestingalternativeespeciallywhenshortanalysistimesarerequired.Forthisreason,mixtureswithawiderangeofcomponentpolarityarebestanalysedbyshortchains,suchasYMC-PackPro C4.

WithintheProFamily,YMC-PackProC4istheselectivityofchoicetoreducetimeofanalysisincombinationwiththegivenadvantagesoftheProFamily,namelythehighpuritysilicasupportandthelowmetalcontent,whichresultinexcellentpeakperformanceasbelow.

1:Phenytoin2:PropranololHCI3:Quinidine4:Lidocaine5:DiltiazemHCI6:VerapamilHCI7:NicardipineHCI

1:Cytosine2:Uracil3:Guanine4:Thymine5:Adenine

Efficient separationof pharmaceutical drugs*

Fast separation of nucleic acid bases*

Antiarrhythmic drugsColumn: YMC-PackProC4(5µm,12nm)150x4.6mmIDPartNo.: BS12S05-1546WTEluent: 20mMKH2PO4-K2HPO4(pH6.9)/methanol(40/60)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat220nm

Nucleic acid basesColumn: YMC-PackProC4(5µm,12nm)150x4.6mmIDPartNo.: BS12S05-1546WTEluent: 20mMKH2PO4

Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat254nm

YMC-Pack Pro C4 SpecificationParticle size / µm 3; 5Pore size / nm 12 Surface area / m2g-1 330Carbon content / % 7Recommended pH range 2.0 - 7.5

YMC ProFamily 73


Column care

YMC-PackProC4isstabletowardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.Cloggedinletfritsoftencanbecleanedbychangingtheflowdirectionorreplacement.

Fordetailedinformationpleaserefertothe“ColumnCareandUseInstructions”,whichareshippedwitheachanalyticalcolumn.

YMC-Pack Pro C4ThecomparisonshownbelowdemonstratesthatYMC-PackProC4isthecolumnofchoicewhenfastHPLCisrequired.ThereisalmostnodifferenceinretentiontimesforthefirstthreecompoundswhilstNicardipineHClelutesfasteronYMC-PackPro C4 due to its lower polarity.


Column: 150x4.6mmIDEluent: 20mMKH2PO4-K2HPO4(pH6.9)/methanolFlowrate: 1.0ml/minTemperature: 37°CDetection: UVat220nm

Optimisation of separation using Pro C8 and Pro C4

Antiarrhythmics1.Phenytoin2.PropranololHCl3.Quinidine4. Lidocaine5.DiltiazemHCl6.VerapamilHCl7.NicardipineHCl

74 YMC ProFamily

YMC-Pack Pro C18 RS

General

The relatively high carbon load ofYMC-Pack Pro C18 RS with 22%amplifies the selectivity’s ability todiscriminate between closely relat-ed compounds such as positionalorsteric isomers.Agoodsystemtotestthisstericselectivityisamixtureofo-,m-andp-terphenylseparatedunder methanol/water conditions.Thesethreecompoundsdifferonlyintheirthree-dimensionalstructureandnotintheirhydrophobicityorpolarity.YMC-PackPro C18RS recognizesevenslightstericdifferencesasshownin the chromatogram on the right,whilst amore conventional carbonload(15%)C18chemistrydoesnot.

- strongly hydrophobic due to carbon content of 22%

- exhibits extraordinary steric selectivity

- extended pH and temperature stability

- for the separation of hydrophobic, acidic and basic molecules

22

USPL1

1-10

Steric selectivity*

1.

2.

3.

m-Terphenyl

o-Terphenyl

p-Terphenyl

1

2

3

Pro C18 RS

10 200 min 10 200 min

Inertsil ODS-3V

1

2,3


1

2

3

Pro C18 RS

10 200 min 10 200 min

Inertsil ODS-3V

1

2,3

*itisrecommendedtouseatleast10%organicsolventcompositionnearthepHlimitsandover50%atpHvaluesabovepH9.0topreservecolumnlifetimes

YMC-Pack Pro C18 RS SpecificationParticle size / µm 3; 5Pore size / nm 8 Surface area / m2g-1 510Carbon content / % 22Recommended pH range 1.0 - 10.0*

YMC ProFamily 75


YMC-Pack Pro C18 RS

Ch

an

ge

of

nu

mb

er

of

the

ore

tic

al

pla

tes

of

Me

thyl

be

nzo

ate

in

%

h100 200 300 400 500 600 7000

20

40

60

80

100

120

Stability under acidic conditions*

Note:WhenassessingpHstabilitydata,pleasetakecaretocertifythatcompletechromatographicconditions are presented.

Stability under basic conditions*

ForpH10,aboratebuffersystem(20mMH3BO3-NaOH(pH9.8)/methanol50/50at30°C)wasselectedtobecontinuouslypumpedthroughthecolumn,whilecheckingthenumberoftheoreticalplatesformethylbenzoateevery50hours.

Full stability

for 3 monthone shift operatio

n!

pH 1

pH 10

The hydrophobicity of Pro C18 RS is hardly changed under the harsh condition (pH: 1, temp.: 70 °C).

Pro C18 RSConventionalODSmonomericODS

Retentionof

Butylbenzene(%)

Basiceluentsmaysignificantlyaffectsilicasandtraditionalbondingchemistries.Therefore,stabilitydatashouldbeconsideredonlyafterverifyingthatthebuffersystemusedmaintainstheselectedpHduringpreparationanduse.Furthermore, itmustbeverifiedthat theeluent isnot recycled,sincethe“active”basicsitesmayequilibratetoasaturationlevelwithtime,resultinginnofurtherinteractionstakingplace.Consequently,onlycontinuousflowof“fresh”andthoroughlybufferedeluentwillprovideaccurateandmeaningfulperformancedata.

76 YMC ProFamily

YMC-Pack Pro C18 RS

10 200 min 10 200 min10 200 min 10 200 min

pH 7 pH 7.5 pH 8 pH 8.5

1

2

1

2

1

2

1

2

k'1 2.14N1 600

k'1 4.15N1 700

k'1 6.50N1 1100

k'1 7.48N1 2200

Erythromycin

1.

O

CH3CH3

OH

HH

CH3

H3C

OCH3OCH3

O

H

O

OHH

OHHOCH3H3C

H3C

H5C2H3C

O

H

H3CH

N

OH

H

OCH3

O

Naphthalene(I.S.)

2.

Method optimization*

10 200 min

20mM37°C

10 200 min

50mM37°C

1

2

1

2

k'1 7.48N1 2200

k'1 7.21N1 2800

50mM45°C

10 200 min

2

1

k'1 8.38N1 5300

Naphthalene(I.S.)

2.

Erythromycin

1.

O

CH3CH3

OH

HH

CH3

H3C

OCH3OCH3

O

H

O

OHH

OHHOCH3H3C

H3C

H5C2H3C

OH

H3CH

N

OH

H

OCH3

O

YMC-Pack Pro C18 RS:Ideal for the separation of steric demanding compounds and/or for useunder broader pH conditions!

Column: YMC-PackProC18RS(5µm,8nm)150x4.6mmIDEluent: potassiumphosphatebuffer(pH8.5)/acetronitrile(50/50)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat210nm

YMC ProFamily 77


YMC-Pack Pro C18 RSApplications

ThespecificpropertiesofYMC-PackProC18RSmakeitanexcellentchoicefortheseparationofnon-polarstructurallyrelatedanalytes.TheextendedresistancetowardsacidicandbasicconditionsnotonlywidensthepossibilitiesinmethoddevelopmentbutalsoprovidesfurtherselectivitiesfordemandingseparationssuchasLC-MSorcombinatorialchemistryof:

positionalisomers,largehydrophobicmolecules,basicandacidiccompounds,peptides

1. [Arg8]-Vasopressin 2. Oxytocin 3. Bradykinin 4. AngiotensinII 5. Leu-Enkephalin 6. AngiotensinI1

2

3

4 6

5

5 10 150 min

1. Flurbiprofen 2. Methylbenzoate 3. Lidocaine 4. DiltiazemHCl 5. VerapamilHCl

1

2

3

4

5

10 20 300 min

Peptides: LC-MS conditions* Drugs: LC-MS conditions*

Nicardipine hydrochloride*Food preservatives* Chlorophenol isomers*

1

2

3

10 200 min

1.

2.

3.

m-Chlorophenol

OH

Cl

o-Chlorophenol

OHCl

OH

Clp-Chlorophenol

1

23

5 10 150 min

1.

Benzoic acid

COOH 2.

Sorbic acid

CH3CH CHCH CHCOOH

3.

Dehydroacetic acid

O

O

O

COCH3

H3C


10 20 30 400 min

degradants

Nicardipine HCl

N

CH2COOCH2CH2N

NO2

H3C

H3COOC

CH3

CH3·HCl

Degradation product

Column: YMC-PackProC18RS(5µm,8nm)150x4.6mmIDPartNo.: RS08S05-1546WTEluent: A:water/TFA(100/0.1) B:acetonitrile/TFA(100/0.1)Gradient: 15-35%B(0-15min)Flow: 1.0ml/minDetection: UV,220nm,0.32AUFSTemperature: 37ºCInjection: 20µl(0.05mg/ml)

Column: YMC-PackProC18RS(5µm,8nm)150x4.6mmIDPartNo: RS08S05-1546WTEluent: 20mMCH3COONH4-NH3(pH8.9)/methanol(35/65)Flow: 1.0ml/minDetection: UV,220nm,0.64AUFSTemperature: 30ºCInjection: 10µl(0.1mg/ml,0.036µl/ml)

Column: YMC-PackProC18RS(5µm,8nm) 150x4.6mmIDPartNo.: RS08S05-1546WTEluent: 20mMKH2PO4-K2HPO4(pH6.9)/ methanol(25/75)Flow: 1.0ml/minDetection: UVat254nm,0.32AUFSTemperature:37ºCInjection: 10µl

Column: YMC-PackProC18RS(5µm,8nm) 150x4.6mmIDPartNo.: RS08S05-1546WTEluent: 20mMCH3COONa-CH3COOH (pH4.3)/acetonitrile(80/20)Flow: 1.0ml/minDetection: UVat230nm,0.26AUFSTemperature:37ºCInjection: 10µl(0.02mg/ml)

Column: YMC-PackProC18RS(5µm,8nm) 150x4.6mmIDPartNo.: RS08S05-1546WTEluent: acetonitrile/water/aceticacid (30/70/1)Flow: 1.0ml/minDetection: UVat230nm,0.32AUFSTemperature:37ºCInjection: 10µl(0.1-0.2mg/ml)

78 YMC ProFamily

Hydrosphere C18- stable under the use of 100% aqueous eluent

- “hydrophilic” C18 surface for enhanced polar recognition

- no need for ion pair reagents

- based on highly inert, ultrapure, pH neutral silica

- specifically designed for pharmaceutical and biotechnology R&D

USPL1

General

Theseparationofpolarcompoundsinmanycasesrequireshighlyaqueousmobilephaseconditionstoachievesufficientretentiononthestationaryphase.ConventionalreversedphaseselectivitiesdonotgivereproducibleresultsundertheseconditionsduemainlytothecollapseoftheC18chains,HydrosphereC18hasbeendeveloped,ontheultrapuresilicasupportoftheProFamily,asthenextgenerationofspecialityphasesfollowingthewellknownYMC-PackODS-AQ,whichwasdevelopedin1987andisstillaveryinterestingselectivityoptionforthesepurposes.

* please be referred to page 52 ff for YMC-UltraHT columns

Hydrosphere C18 SpecificationParticle size / µm 2*; 3*; 5Pore size / nm 12 Surface area / m2g-1 330Carbon content / % 12Recommended pH range 2.0 - 8.0

YMC-Pack ODS-AQ* YMC-Pack Pro C18*

Hydrosphere C18* 1.Toluene2.Testosterone3.n-Propylbenzene4.Acenaphthene5.n-Butylbenzene6.Amitriptyline hydrochlorideexceptional

polarselectivity

symmetrical

peaks even fo

r

strong bases

Column: HydrosphereC18(5µm,12nm)150x4.6mmIDPartNo.: HS12S05-1546WTEluent: 20mMKH2PO4-K2HPO4(pH6.9)/methanol(35/65)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat254nm

+

YMC ProFamily 79


Hydrosphere C18

Its“hydrophilic”C18surfacegivesHydrosphereC18thecapabilitytoshowstableretentiontimesevenafter24hoursunderthesechromatographicconditions.

Hydrosphere C18

Solution: Hydrosphere C18 under 100% aqueous chromatographic conditions*

stableretention!

24 hours

1. Cytosine2.Uracil3.Guanine4.Thymine5. Adenine

Column: HydrosphereC18(5µm,12nm)150x4.6mmIDPartNo.: HS12S05-1546WTEluent: 20mMKH2PO4-K2HPO2(pH6.9)Flowrate: 1.0ml/minTemperature: 37°CDetection: UVat254nm

Afteruse,thecolumnwasstoredovernightwithoutanywashingprocedure.Thenextday,theapplicationwascontinued.

(A) (B)

1. Norepinephrinehydrochloride 2. Epinephine 3. 3,4-Dihydroxyphenylalanine(DOPA) 4. Dopaminehydrochloride 5. 4-Hydroxy-3-methoxymandelicacid 6. 3-Methoxytyraminehydrochloride 7. 4-Hydroxy-3-methoxyphenylglycol 8. 3,4-Dihydroxyphenylaceticacid 9. Homovanilicacid

No ion pair reagent required* Efficient separation of complex substance mixtures*

1. PyridoxineHCl 2. Maleic acid 3. Neostigminemethylsulfate 4. NaphazolineHCl 5. Chlorpheniramine 6. Glycyrrhizin


Ingredients in an eye dropColumn: HydrosphereC18(3µm,12nm)50x4.6mmIDPartNo.: HS12S03-0546WTEluent: A)20mMKH2PO4-H3PO4(pH2.5) B)acetonitrile 0-60%B(0-10min),60%B(10-15min)Flowrate: 1.0ml/minTemperature: 37ºCDetection: UVat265nm,0.128AUFSInjection: 5µl(0.03-0.3mg/ml)

CatecholaminesColumn: HydrosphereC18(5µm,12nm)150x4.6mmIDPartNo.: HS12S05-1546WTEluent: A)50mMKH2PO4-H3PO4(pH3.1) B)50mMKH2PO4-H3PO4(pH3.1)/acetonitrile(90/10) 0%B(0-5min),0-100%B(5-25min,linear),100%B(25-35min)Flow: 1.0ml/minDetection: UVat280nm,0.64AUFSTemperature: 30ºCInjection: 10µl(0.15-0.25mg/ml)

Column care

HydrosphereC18isstabletowardshydrolysisbetweenpH2.0-8.0inupto100%aqueoussystemsandamaximumof50°C.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.


80 YMC ProFamily

Phase dimension

Column ID

[mm]Column length [mm]


[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 AS12S03-H3Q1QT AS12S03-05Q1QT AS12S03-10Q1QT AS12S03-15Q1QT AS12S03-25Q1QT AS12S03-01Q1GC3.0 AS12S03-H303QT AS12S03-0503QT AS12S03-1003QT AS12S03-1503QT AS12S03-2503QT AS12S03-0103GC4.0 AS12S03-H304QT AS12S03-0504QT AS12S03-1004QT AS12S03-1504QT AS12S03-2504QT AS12S03-0104GC4.6 AS12S03-0346WT AS12S03-0546WT AS12S03-1046WT AS12S03-1546WT AS12S03-2546WT AS12S03-0104GC

12 nm5 µm

2.1 AS12S05-H3Q1QT AS12S05-05Q1QT AS12S05-10Q1QT AS12S05-15Q1QT AS12S05-25Q1QT AS12S05-01Q1GC3.0 AS12S05-H303QT AS12S05-0503QT AS12S05-1003QT AS12S05-1503QT AS12S05-2503QT AS12S05-0103GC4.0 AS12S05-H304QT AS12S05-0504QT AS12S05-1004QT AS12S05-1504QT AS12S05-2504QT AS12S05-0104GC4.6 AS12S05-0346WT AS12S05-0546WT AS12S05-1046WT AS12S05-1546WT AS12S05-2546WT AS12S05-0104GC

YMC-Pack Pro C18


Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 OS12S03-H3Q1QT OS12S03-05Q1QT OS12S03-10Q1QT OS12S03-15Q1QT OS12S03-25Q1QT OS12S03-01Q1GC3.0 OS12S03-H303QT OS12S03-0503QT OS12S03-1003QT OS12S03-1503QT OS12S03-2503QT OS12S03-0103GC4.0 OS12S03-H304QT OS12S03-0504QT OS12S03-1004QT OS12S03-1504QT OS12S03-2504QT OS12S03-0104GC4.6 OS12S03-0346WT OS12S03-0546WT OS12S03-1046WT OS12S03-1546WT OS12S03-2546WT OS12S03-0104GC

12 nm5 µm

2.1 OS12S05-H3Q1QT OS12S05-05Q1QT OS12S05-10Q1QT OS12S05-15Q1QT OS12S05-25Q1QT OS12S05-01Q1GC3.0 OS12S05-H303QT OS12S05-0503QT OS12S05-1003QT OS12S05-1503QT OS12S05-2503QT OS12S05-0103GC4.0 OS12S05-H304QT OS12S05-0504QT OS12S05-1004QT OS12S05-1504QT OS12S05-2504QT OS12S05-0104GC4.6 OS12S05-0346WT OS12S05-0546WT OS12S05-1046WT OS12S05-1546WT OS12S05-2546WT OS12S05-0104GC

YMC-Pack Pro C8

Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 BS12S03-H3Q1QT BS12S03-05Q1QT BS12S03-10Q1QT BS12S03-15Q1QT BS12S03-25Q1QT BS12S03-01Q1GC3.0 BS12S03-H303QT BS12S03-0503QT BS12S03-1003QT BS12S03-1503QT BS12S03-2503QT BS12S03-0103GC4.0 BS12S03-H304QT BS12S03-0504QT BS12S03-1004QT BS12S03-1504QT BS12S03-2504QT BS12S03-0104GC4.6 BS12S03-0346WT BS12S03-0546WT BS12S03-1046WT BS12S03-1546WT BS12S03-2546WT BS12S03-0104GC

12 nm5 µm

2.1 BS12S05-H3Q1QT BS12S05-05Q1QT BS12S05-10Q1QT BS12S05-15Q1QT BS12S05-25Q1QT BS12S05-01Q1GC3.0 BS12S05-H303QT BS12S05-0503QT BS12S05-1003QT BS12S05-1503QT BS12S05-2503QT BS12S05-0103GC4.0 BS12S05-H304QT BS12S05-0504QT BS12S05-1004QT BS12S05-1504QT BS12S05-2504QT BS12S05-0104GC4.6 BS12S05-0346WT BS12S05-0546WT BS12S05-1046WT BS12S05-1546WT BS12S05-2546WT BS12S05-0104GC

YMC-Pack Pro C4

Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

8 nm3 µm

2.1 RS08S03-H3Q1QT RS08S03-05Q1QT RS08S03-10Q1QT RS08S03-15Q1QT RS08S03-25Q1QT RS08S03-01Q1GC3.0 RS08S03-H303QT RS08S03-0503QT RS08S03-1003QT RS08S03-1503QT RS08S03-2503QT RS08S03-0103GC4.0 RS08S03-H304QT RS08S03-0504QT RS08S03-1004QT RS08S03-1504QT RS08S03-2504QT RS08S03-0104GC4.6 RS08S03-0346WT RS08S03-0546WT RS08S03-1046WT RS08S03-1546WT RS08S03-2546WT RS08S03-0104GC

8 nm5 µm

2.1 RS08S05-H3Q1QT RS08S05-05Q1QT RS08S05-10Q1QT RS08S05-15Q1QT RS08S05-25Q1QT RS08S05-01Q1GC3.0 RS08S05-H303QT RS08S05-0503QT RS08S05-1003QT RS08S05-1503QT RS08S05-2503QT RS08S05-0103GC4.0 RS08S05-H304QT RS08S05-0504QT RS08S05-1004QT RS08S05-1504QT RS08S05-2504QT RS08S05-0104GC4.6 RS08S05-0346WT RS08S05-0546WT RS08S05-1046WT RS08S05-1546WT RS08S05-2546WT RS08S05-0104GC

YMC-Pack Pro C18 RS

*Guardcartridgeholderrequired,partno.XPGCH-Q1




YMC ProFamily 81


Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 HS12S03-H3Q1QT HS12S03-05Q1QT HS12S03-10Q1QT HS12S03-15Q1QT HS12S03-25Q1QT HS12S03-01Q1GC3.0 HS12S03-H303QT HS12S03-0503QT HS12S03-1003QT HS12S03-1503QT HS12S03-2503QT HS12S03-0103GC4.0 HS12S03-H304QT HS12S03-0504QT HS12S03-1004QT HS12S03-1504QT HS12S03-2504QT HS12S03-0104GC4.6 HS12S03-0346WT HS12S03-0546WT HS12S03-1046WT HS12S03-1546WT HS12S03-2546WT HS12S03-0104GC

12 nm5 µm

2.1 HS12S05-H3Q1QT HS12S05-05Q1QT HS12S05-10Q1QT HS12S05-15Q1QT HS12S05-25Q1QT HS12S05-01Q1GC3.0 HS12S05-H303QT HS12S05-0503QT HS12S05-1003QT HS12S05-1503QT HS12S05-2503QT HS12S05-0103GC4.0 HS12S05-H304QT HS12S05-0504QT HS12S05-1004QT HS12S05-1504QT HS12S05-2504QT HS12S05-0104GC4.6 HS12S05-0346WT HS12S05-0546WT HS12S05-1046WT HS12S05-1546WT HS12S05-2546WT HS12S05-0104GC

Hydrosphere C18


For other dimensions please refer to page 247 *Guardcartridgeholderrequired,partno.XPGCH-Q1

82 Reversed Phase

Reversed Phase 83


Contents

•YMC-PackODS-AQ............................................. 84-87

•YMC-PackODS-A ............................................... 88-89

•YMC-PackODS-AM ............................................ 90-91

•YMC-PackODS-AL ............................................. 92-93

•YMC-PackPolymerC18 ...................................... 94-95

•YMCbasic ........................................................... 96-97

•YMC-PackC8(Octyl) .......................................... 98-99

•YMC-PackPh(Phenyl) .................................... 100-101

•YMC-PackC4(Butyl) ....................................... 102-103

•YMC-PackTMS(C1) ....................................... 104-105

•YMC-PackCN(Cyano) ................................... 106-107

•OrderingInformation ........................................ 108-110

HPLC Columns YMC RP-Classics

HPLC Columns for Reversed Phase Chromatography

InordertosucceedinHPLC,thechoiceoftheoptimalselectivityisessentialtoestablishefficientseparationconditions.Thebestsuitedpackingmaterialdependssignificantlyonthecharacteristicsoftheseparationconditions,whichshouldbethoroughlyconsidered.

ForthispurposeYMCoffersawidevarietyofselectivitiesapplicabletoHPLCfromnano-scaleanalysistolargescaleisolation.Within this catalogue theworld renownYMC-PackODS-Series (YMC-PackODS-AQ,YMC-PackODS-A,YMC-PackODS-AM,YMC-PackODS-AL)andotherphasesaredescribed.

Introduction

84 Reversed Phase

General

YMC-PackODS-AQ is aC18 reversedphase silica basedHPLCpackingmaterial specificallydesignedforusein100%aqueouseluents.Asaresultoftheproprietaryderivatisationprocess,YMC-PackODS-AQexhibitsadifferentselectivitytothatoftraditionalC18stationaryphases.ThisdifferenceinselectivityofYMC-PackODS-AQcanbeusedtoadvantageforHPLCseparations,whicharedifficulttoachievewithconventionalC18columns. Selectivity Data

TheproprietaryYMCderivatisationprocesscreatesthedifferentselectivityofYMC-PackODS-AQ,where:

1.Theactivityofacidicunreactedsilanolsisreduced,allowingmoderatelybasiccompoundstobeelutedwithlittleornopeaktailing.

2.Thebalancedhydrophilic/lipophilicnatureoftheYMC-PackODS-AQstationaryphaseleadstostrongretentionsofpolarsamplesoluteseveninaqueouseluents.

ThesepropertiesofYMC-PackODS-AQarebeneficialforseparationsofpolarorganiccompounds,whichtendnottoberetainedorareunresolvedwhenconventionalC18columnsareused.

ManyconventionalODSpackingslosetheirabilitytoretainpolarcompoundsinthesehighaque-ouscontentmobilephasesasshownopposite.TheyappearlesslipophilicwithdenselyfoldedC18chains.However,insimilarmobilephases,YMC-PackODS-AQmaintainsitsbrush-likeC18chainstructureanditslipophilicpropertiesandprovidesexcellentretentionofpolarcompounds.

Applications

YMC-PackODS-AQisabletoresolvecompoundswithminordifferencesinpolarityfromcloselyrelatedchemicalstructures.Asaresult,YMC-PackODS-AQisanexcellenttoolfortheseparationofdrugsandcorrespondingmetabolites,pesticidesanddegradationproducts,orpeptidesandproteindigestsetc.Thiscapabilityof“polarrecognition”opensupabroadapplicationrangeforYMC-PackODS-AQinlifesciencesandpharmacology.

Genuinelinearscale-upfromanalyticaltolargescaleseparationsiseasilyachievablewithYMCproductssuchasYMC-PackODS-AQ,whereparticlesizesfrom3to150µmareavailableinlargelotsizesuptoseveralhundredkilograms,ifneeded.This,togetherwiththeoutstandingselectivityofYMC-PackODS-AQ,makeitanessentialtooltoenhancetheproductivityoflargescalechro-matographicprocesses.

YMC-PackODS-AQisalsoavailableinpreparativeparticlesizes.

- “hydrophilic” C18- balanced surface chemistry- polar recognition- metabolite recognition

YMC-Pack ODS-AQ

YMC-Pack ODS-AQ Specification

Particle size / µm 3; 5 3; 5Pore size / nm 12 20Surface area / m2g-1 330 175Carbon content / % 14 10Recommended pH range 2.0 - 7.5 2.0 - 7.5

USPL1

Reversed Phase 85


YMC-Pack ODS-AQComparison of ODS-AQ vs. Conventional ODS

ConventionalODS-Column

YMC ODS-AQ

Initial

After24 hoursof flow

After1 hourof flow

Initial

=C18 =Endcapping = Water

=C18 =Endcapping = Water

Conventional ODS instandard organic mobile phases*

Conventional ODS in100% aqueous mobile phases*

YMC-Pack ODS-AQ instandard organic mobile phases*

YMC-Pack ODS-AQ in100% aqueous mobile phases*

Column: YMC-PackODS-A(25µm,12nm)250x10mmIDMobilePhase:100%waterCompounds: Highlypolarandverywatersoluble

Column: YMC-PackODS-AQ(25µm,12nm)250x10mmIDMobilePhase: 100%waterCompounds: Highlypolarandverywatersoluble

86 Reversed Phase

YMC-Pack ODS-AQ

1:Noradrenaline 2:Adrenaline 3:DL-3,4-DihydroxymandelicAcid 4:3,4-Dihydroxyphenylalanine(DOPA) 5:DopamineHydrochloride 6:Tyrosine 7:DL-3-Methoxy-4-HydroxymandelicAcid 8:Phenylalanine 9:4-Hydroxy-3-MethoxyPhenylglycol10:5-Hydroxyindole-3-AceticAcid11:VanillicAcid12:HomovanillicAcid

| | | | 0 10 20 30 min

2

34

56

7

9

12

1

108

11

CatecholaminesColumn: YMC-PackODS-AQ(5µm,12nm)250x4.6mmIDPartNo.: AQ12S05-2546WTEluent: A:phosphatepuffer(100mM,pH3.0)B:acetonitrileGradient: 99%A(0-20min),85%A(20-25min)Flow: 1.5ml/minDetection: UVat210nmTemperature: Roomtemperature

1: Glycolicacid 2: L-Malicacid 3: Malonicacid 4: Lacticacid 5: Aceticacid 6: Maleicacid

7: Citricacid 8: Fumaricacid 9: Succinicacid10: Propionicacid

Exceptional performance for the separation of polar compounds*

No need for ion pair reagents*Strong retention in aqueous eluents*

Cephalosporin antibioticsColumn: YMC-PackODS-AM(5µm,12nm)150x4.6mmIDPartNo.: AM12S05-1546WTEluent: methanol/water/aceticacid(10/85/5)Flow: 1.0ml/minDetection: UVat260nm,0.16AUFSTemperature: 37°C

Cephalosporin antibioticsColumn: YMC-PackODS-AQ(5µm,12nm)150x4.6mmIDPartNo.: AQ12S05-1546WTEluent: methanol/water/aceticacid(10/85/5)Flow: 1.0ml/minDetection: UVat260nm,0.16AUFSTemperature: 37°C

Crude drugsColumn: YMC-PackODS-AQ(5µm,12nm)250x4.6mmIDPartNo.: AQ12S05-2546WTEluent: 20mMH3PO4-NaH2PO4(pH2.8)Flow: 0.7ml/minDetection: UVat220nm,0.08AUFSTemperature: 30°CInjection: 10µl(0.007~1.8mg/ml)

Conventional ODS

YMC-Pack ODS-AQ

51015min 5101520min

Reversed Phase 87


YMC-Pack ODS-AQ

1: Cytosine 2: Cytidine 3: Uracil 4: Deoxycytidine 5: Guanine 6: Adenine 7: Hypoxanthine 8: Uridine 9: Xanthine10: Thymine11: Inosine12: Guanosine13: Deoxyinosine14: Deoxyguanosine15: Xanthosine16: Thymidine17: Adenosine18: Deoxyadenosine

BycourtesyofDr.ManfredRaida,IPFHannover

1:Arbutin 2: Salicin 3: Esculine 4: g-Strophanthin (Quabain) 5: Baicalin 6: Barbaloin 7: Glycyrrhizin 8: Digitoxin

Comparison of YMC-Pack ODS-AQ with competitive products

Separation of biomolecules* Excellent choice for a broad chromatographic appli-cation range*

YMC-PackODS-AQ

Since1987,YMC-PackODS-AQhasconsistentlyincreaseditspopularityduetoitsnovelselectivitypatterntowardspolarcompoundsanditsabilitytowithstand100%aqueousconditions.Today,morethan25(!)yearslater,manynewanalyticalandpreparativemethodsarestillbeingdevelopedonYMC-PackODS-AQchemistrydespiteofvariousAQ-typeproductsrecently beingintroducedbyourcompetitors;phaseswithsupposedly“identical”selectivityorwithexoticbondingtechniquesdesignedtogenerateperformancecharacteristicssimilartothoseofYMC-PackODS-AQ.However,genuineYMC-PackODS-AQstillrepresentstodayafullycompetitivestate-of-the-arthighperformancestationaryphase,despitethecomplementaryYMCinnovation,namelynewgenerationHydrosphereC18,describedonpages78-79,asapotentialin-housecompetitor.

ReproSil-Pur C18-AQ

Tryptic digest of BSA


Column care

TherecommendedpHrangeforYMC-PackODS-AQis2.0-7.5inupto100%aqueoussystemsandamaximumof50°C.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.Cloggedinletfritsoftencanbecleanedbychangingtheflowdirection.


| | | | 0 10 20 30 min

| | | | | 0 10 20 30 40 min

NucleosidesColumn: YMC-PackODS-AQ(5µm,12nm)250x4.6mmIDPartNo.: AQ12S05-2546WTEluent: A=20mMCH3COOH-CH3COONH4(pH3.5) B=20mMCH3COOH-CH3COONH4(pH3.5)/methanol=90/10(v/v)Gradient: 30%B(0-5min),30-100%B(5-13min,linear),100%B(13-40min)Flow: 0.7ml/minDetection: UVat260nmTemperature: 30ºC

Crude drugsColumn: YMC-PackODS-AQ(5µm,12nm)250x4.6mmIDPartNo.: AQ12S05-2546WTEluent: A:methanol/NH4H2PO4(20mM)=5/95 B:methanol/NH4H2PO4(20mM)=80/20Gradient: 0-100%B(0-20min,linear),100%B(20-40min)Flow: 0.6ml/minDetection: UVat250nmTemperature: 30°C

niceresolution

88 Reversed Phase

General

YMC-PackODS-A,YMC’sclassicalreversedphasepackingmaterial,isrenownedworldwidebe-causeofitsuniqueperformanceandreproducibility.Duetothehighquality,YMC-PackODS-AiswidelyusedforthevalidationofanalyticalHPLCmethodsandforlong-termreproducibleprepara-tiveHPLCprocesses.

Properties

The production of the base silica forYMC-PackODS-Aand its subsequentderivatisationareperformedinlargebulkbatches.Exhaustiveendcappingreduc-esreliablytheactivityofsilanolgroupsandminimises nonspecific secondaryretention.

Inadditiontostandardmethods,likede-terminationofadsorptionisotherms,par-ticlesizedistributionandcarboncontent(seetableabove),YMCusesanextensiverangeof analyticalmethods to ensureconstantandreproducibleselectivityofthereversedphasepackings.

ThebaseofYMC-PackODS-AisYMC’shigh purity silica. This premium silicacontains only very low levels ofmetalcontaminants and so prevents signifi-cant tailing of samplemolecules suchas8-hydroxyquinolineoracetylacetone,which easily form coordination com-plexeswithmetalionsonthesilicasur-face.Ascoordinatingfunctionalgroupsare frequent structural components inpharmaceuticalcompounds,highpuritypackings such as YMC-PackODS-Aareneededforreproducibleseparationofthesecompoundswithoutsecondaryretentionortailing.

YMC-PackODS-A is also available inpreparativeparticlesizes.

- fully endcapped C18 material

- highly versatile ODS phase

- for polar to moderately nonpolar pharmaceuticals, organic chemicals, biologicals and natural products

YMC-Pack ODS-A* Silica Basewith MetalImpurities*

a bColumn: YMC-PackODS-A(12nm,5µm,150x4.6mmID)Eluent: KH2PO4(20mM,pH7.6)/methanol=40/60Flow: 1.0ml/minDetection: UV,254nmTemperature: 37°CSubstances: 1.Uracil 2.Acetylacetone 3.8-Hydroxyquinoline

Coordination compoundsa: High purity packing material YMC-Pack ODS-Ab: Competitive product

12

3

1

2

3

YMC-Pack ODS-A

YMC-Pack ODS-A Specification

Particle Size / µm 3; 5 3; 5 5Pore Size / nm 12 20 30Surface area / m2g-1 330 175 100Carbon content / % 17 12 7Recommended pH range 2.0 - 7.5 2.0 - 7.5 2.0 - 7.5

USPL1

Reversed Phase 89


YMC-Pack ODS-A

| | | | | 0 5 10 15 20 min

1

2

3

4

5

6

7

89

10

11

12

1: Olaquindox 2: Carbadox 3: Thiamphenicol 4: Sulfamerazine 5: Sulfadimidine 6: Sulfamonomethoxine 7: Furazolidone 8: OxolinicAcid 9: Sulfaquinoxaline10: Sulfadimethoxine11: NalidixicAcid12: PiromidicAcid

| | | | 0 10 20 30min

1: Hydrocortisone 2: Cortisone 3: Methylprednisolone 4: Betamethasone 5: Dexamethasone 6: Corticosterone 7: Beclomethasone 8: Triamcinolone Acetonide

Column: YMC-PackODS-A(5µm,12nm)250x4.6mmIDPartNo.: AA12S05-2546WTEluent: A:ACN/NH4H2PO4(50mM)=10/90 B:ACN/NH4H2PO4(50mM)=80/20Gradient: 25%B(0-5min),25-100%B(5-15min), 100%B(15-20min)Flow: 1.0ml/minDetection: UV,240nmTemperature: 37ºC

Adrenocorticosteroids*Antibacterial agents*

Column: YMC-PackODS-A(5µm,12nm)150x4.6mmIDPartNo.: AA12S05-1546WTEluent: ACN/H2O=27/73Flow: 1.0ml/minDetection: UV,260nmTemperature: 37ºC

Column Care

TherecommendedpHrangeforusingYMC-PackODS-Acolumnsis2.0-7.5.Removeacidandbuffersaltsbeforestor-age.Storethecolumninmethanol/water=50/50.Ifcolumnsareaffectedbyundesiredcontaminantsorcloggedinletfritswhichcausebackpressureincreases,flushthecolumnwithTHFintheoppositeflowdirection.


Applications

YMC-PackODS-Ais frequentlyusedforpharmaceutical,biochemicalandenvironmentalapplicationsaswellasforseparationsinthefieldoffoodtechnology.

YMC-PackODS-Aisavailableinparticlesizesfrom3to50µm.Astheselectivityisidenticalthroughoutthewholerange,thesephasesareidealforscale-upfromanalyticaltopreparativeprocessscale.

Column: YMC-PackODS-A(5µm,12nm)250x4.6mmIDPartNo.: AA12S05-2546WTEluent: 20mMtriethylamine-aceticacid(pH5.7)/acetonitrile(9/1)Flowrate: 1.0ml/minDetection: UVat260nm,0.16AUFSTemperature: 37ºCInjection: 5µl(0.27~0.71mg/ml)

Column: YMC-PackODS-A(5µm,12nm)150x4.6mmIDPartNo.: AA12S05-1546WTEluent: A:water B:methanol 2%B(0-5min),2-30%B(5-15min,linear), 30-100%B(15-20min,linear),100%B(20-30min)Flowrate: 1.0ml/minDetection: EvaporativeMassAnalyzer(ACSLtd.) Temp 150 TimeConstant 5 PMT 3 Attenuation 16 Pressure 20psiTemperature: 30ºCInjection: 200µl(1.5~15mg/ml)

Sugar nucleotides* Cyclodextrins*

102030min

51015202530min

1. g-Cyclodextrin 2.a-Cyclodextrin 3. b-Cyclodextrin 4. O-Dimethyl-b-Cyclodextrin 5. O-Trimethyl-b-Cyclodextrin

1. CDP-D-glucose 2. UDP-D-glucose 3. UDP-N-acetyl-D-glucosamine 4. GDP-D-mannose 5. ADP-D-glucose

90 Reversed Phase

General

ValidationandregistrationofanalyticalHPLCmethodsrequiresthelongtermreproducibilityoftheentireanalyticalprocess.ThehighconsistencyinthequalityofHPLCpackingsandcolumnsplaysakeyroleforvalidatedHPLCanalysis.Therefore,YMCcreatedODS-AM,ahighqualityreversedphaseC18HPLCpackingmaterial tomeet themoststringentdemands forvalidatedanalyticalHPLCprocesses.

Properties

YMC-PackODS-AMisproducedinlargelotsusinghighpurityYMCsilicaasabasematerialandamultistagesynthesisprocess.Forthederivatisation,monomericbondingchemistryisappliedfollowedbyanextensiveendcappingprocesstoreducethesilanolactivity.

The resultingYMC-PackODS-AMpacking isextensively tested to ensure compliancewithspecificationssetforverylowvariationsinphysi-cochemicalproperties.

Inaddition,YMC-PackODS-AMpackingsandcolumns have to pass numerous proprietarychromatographicteststomeetthenarrowqualityspecificationrangewithregardto:

-selectivitypattern-columnresolution-absoluteretention-peaksymmetry YMCappliesvariousteststoperformstatisticalqualitycontrolforreversedphaseHPLCpack-ings.Thea-valuetestofmethylparabenand2,6-dimethylpyridineforinstance,isverysensitiveandisroutinelyusedtomonitortheretentionandtheselectivitypropertiesofYMC-PackODS-AM.

Methylparabenisamoderatelypolar, inertcompound.It isretainedsolelybyaRPmechanism,withminimalsecondary interactionswithresidualsilanolgroups.2,6-dimethylpyridine,however,representsalipophilicaminecompoundwhichhasahighpotentialofunspecificinteractionwithunreactedacidicsilanols.Anincreaseinretentionof2,6-dimethylpyridineandhencelowera-valueswouldindicateincompleteC18bondingand/orineffectiveendcapping.YMCspecifiesforODS-AMthatthestatisticala-valueofmethylparabenand2,6-dimethylpyridinebe2.77+/-0.20.TherigorousqualitycontrolandthequalityassurancesystemappliedbyYMCminimisesthevari-ationinretentionandselectivityofYMC-PackODS-AMcolumns.

Duetotheguaranteedlongtermreproducibility,YMC-PackODS-AMcolumnsoftenarethefinalchoiceforestablishingvalidatedHPLCanalysis.

- high quality analytical C18- tightly specified- long term reproducibility- for method validation- for method registration

a: Methylparaben/2,6-Dimethylpyridine as reference

Statistical Quality Control (a-value monitoring)

a

Consecutive Lots

YMC-Pack ODS-AM

YMC-Pack ODS-AM Specification

Particle Size / µm 3; 5Pore Size / nm 12Surface area / m2g-1 330Carbon content / % 17Recommended pH range 2.0 - 7.5

USPL1

Reversed Phase 91


YMC-Pack ODS-AMApplications

ODS-AMhasanappropriateselectivityforpolartomoderatelynonpolarpharmaceuticals,organicintermediates,biologi-calandnaturalproductsfoundinthechemicalandpharmaceuticalindustry.

Nucleotides*

Column: YMC-PackODS-AM(5µm,12nm)250x4.6mmIDPartNo.: AM12S05-2546WTEluent: A=0.2Mtriethylamine-aceticacid(pH6.6) B=0.2Mtriethylamine-aceticacid(pH6.6)/ACN(95/5)Gradient: 4%B(0-10min),4-100%B(10-35min,linear), 100%B(35-50min)Flow: 1.0ml/minDetection: UVat260nm,0.16AUFSTemperature: 30°CInjection: 14µl(0.029~0.16mg/ml)

Column: YMC-PackODS-AM(5µm,12nm)250x4.6mmIDPartNo.: AM12S05-2546WTEluent: A=60mMCH3COONa-CH3COOH(pH6.6)/ACN(94/6) B=60mMCH3COONa-CH3COOH(pH6.6)/ACN(40/60)Gradient: 0-55%B(0-20min,linear),100%B(20-25min),0%B(25-35min)Flow: 1.0ml/minDetection: UVat250nm,0.08AUFSTemperature: 37ºCInjection: 5µl(62.5pmol/µl)Sample: PTCderivatives

PTC-amino acids*

1: L-Asparticacid 2: L-Glutamicacid 3: L-Serine 4: Glycine 5: L-Histidine 6: L-Arginine 7: L-Threonine 8: L-Alanine 9: L-Proline

10: NH311: L-Tyrosine12: L-Valine13: L-Methionine14: L-Cystine15: L-Isoleucine16: L-Leucine17: L-Phenylalanine18: L-Lysine

Column Care

TherecommendedpHrangeforusingYMC-PackODS-AMcolumnsis2.0-7.5.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=50/50.Ifcolumnsareaffectedbyundesiredcontaminantsorcloggedinletfritswhichcausebackpressureincreases,flushthecolumnwithTHFintheoppositeflowdirection.


10203040min

1: 5‘-CMP 2: 5‘-UMP 3: 5‘-dCMP 4: 5‘-CDP 5: 5‘-GMP 6: 5‘-UDP 7: 5‘-IMP 8: 5‘-CTP 9: 5‘-UTP10: 5‘-GDP11: 5‘-TMP12: 5‘-AMP13: 5‘-dGMP14: 5‘-GTP15: 5‘-TDP16: 5‘-ADP17: 5‘-TTP18: 5‘-dAMP19: 5‘-ATP

51015202530min

92 Reversed Phase

YMC-Pack ODS-AL

General

YMC-PackODS-ALusesnotonlyhydrophobic interactionbutalsosecondary interactionswithreactiveresidualsilanolgroupstoaffectseparation.Thisresultsinadifferentselectivityfromcon-ventionalODScolumns.Whenionicinteractionsareinvolved,itispreferabletouseabufferinthemobilephasetoachievereproducibleseparations.

- residual silanols for mixed-mode separations

- same high Iigand density as other YMC ODS phases

- unique selectivity for polar compounds

YMC-Pack ODS-AL Specification

Particle Size / µm 3; 5Pore Size / nm 12Surface area / m2g-1 330Carbon content / % 17Recommended pH range 2.0 - 7.5

Effects of silanol groups*

YMC-Pack ODS-AL YMC-Pack ODS-AM

1:Uracil2:Pyridine3:2,6-diemetylpyridine4:Methylparaben

Theseparationfactor(a)ofinternalstandardsmethylparaben/2,6-dimethylpyridineforYMC-PackODS-AL,whichisnotendcappedisdifferenttothatofYMC-PackODS-AM.Duetotheresidualsilanolgroups,YMC-PackODS-ALshowsrelativelygreatretentionofpyridines.

USPL1

Reversed Phase 93


YMC-Pack ODS-AL

Steroids from toad venom*

Column care

TherecommendedpHrangeforusingYMC-PackODS-ALcolumnsis2.0-7.5.Removeacidandbuffersaltsbeforestor-age.Storethecolumninmethanol/water=50/50.Ifcolumnsareaffectedbyundesiredcontaminantsorcloggedinletfritswhichcausebackpressureincreases,flushthecolumnwithTHFintheoppositeflowdirection.


Column: YMC-PackODS-AL(5µm,12nm)150x4.6mmIDPartNo.: AL12S05-1546WTEluent: 100mMKH2PO4-Na2HPO4(pH7.0)/methanol(75/25)Flow: 0.8ml/minDetection: UVat270nm,0.13AUFSTemperature: 30ºCInjection: 3µl(0.1~0.8mg/ml)

Column: YMC-PackODS-AL(5µm,12nm)150x4.6mmIDPartNo.: AL12S05-1546WTEluent: ACN/0.1%phosphoricacid(45/55)Flow: 1.0ml/minDetection: UVat300nm,0.08AUFSTemperature: 40°CInjection: 10µl(0.04~0.08mg/ml)

Applications

Vitamin D2 and Vitamin D3*Sterols*

Column: YMC-PackODS-AL(5µm,12nm)150x4.6mmIDPartNo.: AL12S05-1546WTEluent: acetonitrile/water(95/5)Flow: 1.0ml/minDetection: UVat265nmTemperature: 40ºCInjection: 10µl(0.01mg/ml)

Column: YMC-PackODS-AL(5µm,12nm)250x4.6mmIDPartNo.: AL12S05-2546WTEluent: methanol/water(100/2)Flow: 0.8ml/minDetection: UVat210nm,0.32AUFSTemperature: 37°CInjection: 10µl(0.4~1mg/ml)

51015min

10203040min

Disinfectants*

94 Reversed Phase

- hydrophilic polymethacrylate support

- excellent reproducibility of C18 chemistry integral to polymer matrix

- no silanol or metal contaminants

- pH stable from pH 2 - 13

- compatible with all standard reversed phase solvents

YMC-Pack PolymerC18

YMC-Pack PolymerC18 Specification

Particle Size / µm 6Pore Size / nm proprietarySurface area / m2g-1 n/aCarbon content / % 10Recommended pH range 2.0 - 13.0

2-13

General

YMC-PackPolymerC18isareversedphaseliquidchromatographypackingwhichprovidesabroadrangeofsolventchoicesandapHrangefrom2.0-13.YMC-PackPolymerC18ismanufacturedfromahydrophilicmethacrylatepolymerwhichiscross-linkedwithC18ligand-containingreagents.YMC-PackPolymerC18offersamaximumapplicationrange:awidevarietyofcompoundssuchasorganicacids,organicamines,peptides,pharmaceuticalsandproteinscanbeseparatedusingYMC-PackPolymerC18.

Properties

YMC-PackPolymerC18ispreparedfromahydrophilicmethacrylatepolymerbondedwithahydro-phobicoctadecylsilanereagenttomaketheC18functionalityanintegralpartofthepolymericstruc-ture.Thisgivesathree-dimensionalpolymermatrixwhichisnotbasedonasilicagelsupport.

Assuch, ithasno residualsilanolsormetal impurities to interferewith theseparationofbasicorganiccompounds.

YMC-PackPolymerC18 is compatiblewith all common reversedphase eluents suchaswater,methanol,acetonitrileandTHF.Virtuallyallaqueousbuffersandacidmodifiers,suchasTFAandphosphoricacid,aswellasbasemodifierssuchassodiumhydroxideandammoniumhydroxidecanbeused.Sinceitresistsshrinkingandswelling,YMC-PackPolymerC18canbeusedwitheluentsrangingincompositionfrom100%aqueousto100%organiccomponent.

Inaddition,YMC-PackPolymerC18caneasilybesterilisedbyflushingwith0.1MNaOHin20%acetonitrile/water.

TheselectivityandretentionofYMC-PackPolymerC18issimilartostandardODSphases,duetoitshydrophobicbondingonahydrophilicsupport.Consequently,itsselectivityisclosertothatofsilica-basedC18supportsthantostyrene/DVB-basedsupports.

Itshouldbenotedthatinteractionsbetweenaromaticorconjugatedsystemsandthemethacrylatebackboneprovidesslightlygreaterretentionwhencomparedtosilica-basedODScolumns,whereashighlyaliphaticcompoundsshowgreaterretentiononsilica-basedODSsupports.

YMC-PackPolymerC18isalsoavailableinpreparativeparticlesizes.

Reversed Phase 95


YMC-Pack PolymerC18

Aminoalcohols*

Column: YMC-PackPolymerC18250x6.0mmIDPartNo.: PC99S06-2506WTEluent: 100mMNa2HPO4/100mMNaOH(60/40,pH12.0)Flow: 0.6ml/minDetection: UVat215nm,0.32AUFSTemperature: 20°CInjection: 50µl(0.2~3.0mg/ml)

Peptides and proteins*

Column: YMC-PackPolymerC18150x4.6mmIDPartNo.: PC99S06-1546WTEluent: A=acetonitrile/water/TFA(20/80/0.05) B=acetonitrile/water/TFA(45/55/0.05) 0-100%B(0-30min,linear)Flow: 1.0ml/minDetection: UVat220nm,0.32AUFSTemperature: 30ºCInjection: 30µl

1: Met-Enkephalin 2: Leu-Enkephalin 3: BSA 4:a-Matingfactor 5: Insulin 6: Ovalbumin

Column care

YMC-PackPolymerC18isstabletowardshydrolysisbetweenpH2.0-13.0.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.Ifcolumnsareaffectedbyundesiredcontaminantsorcloggedinletfritswhichcausebackpressureincreases,flushthecolumnwithTHFintheoppositeflowdirection.


Applications

Organic acids*

Column: YMC-PackPolymerC18250x4.6mmIDPartNo.: PC99S06-2546WTEluent: 0.1%TFAFlow: 0.5ml/minDetection: UVat220nm,0.08AUFSTemperature: 30°CInjection: 10µl(0.016~2.2mg/ml)

1020min

1: Ethanolamine 2: Diethanolamine 3: N-Methylethanolamine 4: Triethanolamine 5: N,N-Dimetylethanolamine

5101520253035min

1. L-Mailicacid 2. Lacticacid 3. Aceticacid 4. Maleic acid 5. Succinicacid 6. Propioniacid 7. Fumaricacid

96 Reversed Phase

YMCbasic- alternative bonding approach to reduce peak tailing of basic pharmaceuticals

- no need for ion pair reagents or amine modifiers

- complementing selectivity to C8 and C18 materials

YMCbasic Specification

Particle size / µm 3; 5Pore size / nm proprietarySurface area / m2g-1 proprietaryCarbon content / % 7.5Recommended pH range 2.0 - 7.5

General

The proprietary derivatisation pro-cedureforYMCbasicallowsYMCtoproduce amaterialwith controlledsurface coverage,which showsex-cellent lot-to-lot reproducibilityasaresultofcloselymonitoringboththeproductionofthesilicasupportandthebondingprocess.

The resulting YMCbasic materialshowsadifferent hydrophobicity toC8orC18phases as shown in thediagramonthispage.Finally, it rep-resents an interesting alternative toshortchainselectivities.

Applications

The result is a phasewith true re-versed phase characteristics, highresolution and excellent peak sym-metry forbasiccompoundswithouttheneedforionpairreagentsoraminemodifiers(seeseparationofanilinesusingacetonitrile/watereluent).Un-likemanybase-deactivatedphases,YMCbasicisalsosuitableforseparationofacidiccompounds,showingslightretentionofhighlypolaracidcompoundssuchasmaleate.YMCbasicprovidesacomplementingselectivityseenwithconventionalC8andC18materials,butwithoutpeaktailingforbasiccompounds.

YMCbasicisalsoavailableinpreparativeparticlesizes.

YMCbasicYMC-Pack OctylYMC-Pack ODS-AM

Hydrophobicity of YMCbasic versusYMC-Pack Octyl and YMC-Pack ODS-AM*

Column: 250x4.6mmIDEluent: acetonitrile/water(75/25)Flow: 1.0ml/minDetection: UVat254nm

5.12

5 To

luen

e

5.79

2 E

thyl

ben

zene

6.80

8 P

rop

ylb

enze

ne

8.16

7 B

utyl

ben

zene

7.20

0

12.1

25

USPL7

Reversed Phase 97


YMCbasic

1. Phenol 2. Dimethylaniline 3. Toluene 4. Diethylaniline

Anilines and aromatic compounds* Basic compounds*

1. Maleate 2. Phenylpropanolamine 3. Pseudoephedrine 4. Scopalamine 5. Chlorpheniramine

Cough/cold medicine*

1. Uracil 2. Procainamide 3. N-Acetylprocainamide 4. 4-Hydroxyacetophenone


Column care

TherecommendpHrangeforYMCbasicis2.0-7.5.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.Cloggedinletfritsoftencanbecleanedbychangingtheflowdirectionorreplacement.


Column: YMCbasic(5µm)250x4.6mmIDPartNo.: BA99S05-2546WTEluent: acetonitrile/water(60/40)Flow: 2.0ml/minDetection: UVat254nm

Column: YMCbasic(5µm)250x4.6mmIDPartNo.: BA99S05-2546WTEluent: acetonitrile/0.05mMKH2PO4(pH3.2)(15/85)Flow: 2.0ml/minDetection: UVat254nm

Column: YMCbasic(5µm)150x4.6mmIDPartNo.: BA99S05-1546WTEluent: 20mMK2HPO4-KH2PO4(pH6.9)/acetonitrile(92/8)Flow: 1.0ml/minDetection: UVat254nm,0.32AUFSTemperature: 37°CInjection: 10µl(30~300µg/ml)

1.534.56min

2468min

102030min

98 Reversed Phase

- alternative phase to C18 with moderate hydrophobicity

- fully endcapped, high coverage monomeric bonded chemistry

- ideal for method development and routine separations

- excellent retention for all types of organic molecules, especially peptides, proteins and pharmaceuticals

YMC-Pack C8 (Octyl)

YMC-Pack C8 Specification


General

YMC-PackC8isoneofYMC’smostcommonlyusedbondedphasesandanexcellentalternativetoC18selectivities.Duetoitsmoderatehydrophobicity,retentiontimestendtobeshorterthanthoseforODSphases.YMC-PackC8issuitableforawiderangeofsampletypesincludingpharmaceu-ticalsandbiologicalswitharelativelyhighhydrophobicity.

Properties

YMC-PackC8ispreparedbyexhaustivebondingofamonomericoctylsilanetototallysphericalandporoussilicagel.Thebondedphaseisthentreatedwithanexhaustiveendcappingprocesstoensureahighsurfacecoverageleadingtoamoderate10%carbonloadingonthestandard12nmporematerial.ComparedtoC18phases,retentiontimesforhydrophobicmoleculeswillbeshorteronC8materialduetothereducedcarbonload.

YMC-PackC8isideallysuitedfortheseparationofmanycompoundsthataretoostronglyretainedonC18phasesorwhichrequiregreaterretentionthanprovidedbyC4materials.Itisoneofthemostversatile reversedphasematerialsandshouldbeconsidered for thedevelopmentofnewmethods.

Available in threeporosities,YMC-PackC8materialwill separatemanyclassesof compoundsincludingpharmaceuticals,organicchemicals,peptides,proteinandotherbiologicalmolecules.Forpreparativeapplications,choosethesmallestporesizewhichprovidesadequateretentionandresolution.Thisisbecausesampleloadingisgenerallyproportionaltosurfacearea.Smallerporositymediaprovidegreatersurfaceareaandhencegreaterloadability.

YMC-PackC8(Octyl)isalsoavailableinpreparativeparticlesizes.

Column care

YMC-PackC8(Octyl)isstabletowardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.Ifcolumnsareaffectedbyundesiredcontaminantsorcloggedinletfritswhichcausebackpressure increases,flushthecolumnwithTHFintheoppositeflowdirection.Fordetailedinformationpleaserefertothe“ColumnCareandUseInstructions”whichareshippedwitheachanalyticalcolumn.

USPL7

Reversed Phase 99


YMC-Pack C8 (Octyl)

Column: YMC-PackC8(Octyl)(5µm,12nm)250x4.6mmIDPartNo.: OC12S05-2546WTEluent: acetonitrile/THF/water(46/4/50)Flow: 0.7ml/minDetection: UVat230nm,0.16AUFSTemperature: 30ºCInjection: 7µl(0.1mg/ml)

Nicotinic acid analogues*Estrogens and progestins*

Column: YMC-PackC8(Octyl)(5µm,12nm)150x4.6mmIDPartNo.: OC12S05-1546WTEluent: acetonitrile/20mMKH2PO4(5/95)Flow: 1.0ml/minDetection: UVat260nm,0.64AUFSTemperature: 30ºCInjection: 13µl(0.2mg/ml)

Applications

Column: YMC-PackC8(Octyl)(5µm,12nm)150x4.6mmIDPartNo.: OC12S05-1546WTEluent: methanol/100mMKH2PO4(10/90)Flow: 0.8ml/minDetection: UVat285nm,0.16AUFSTemperature: 37ºCInjection: 10µl(0.05~0.4mg/ml)

Resorcylic acid isomers*

204060min

510min

102030min

100 Reversed Phase

- fully endcapped, monomeric phenyl phase

- unique selectivity due to π - π interactions

- preferential retention of aromatic compounds

- alternative selectivity to C18, C8 or C4 bonded phases for the analysis of peptides and other biomolecules

YMC-Pack Ph (Phenyl)

YMC-Pack Ph Specification

Particle Size / µm 3; 5 5Pore Size / nm 12 30Surface area / m2g-1 330 100Carbon content / % 9 3Recommended pH range 2.0 - 7.5 2.0 - 7.5

USPL11

General

YMC-PackPh(Phenyl)isahighdensitybondedphase(9%carbonloadon12nmsilica)whichisfullyendcapped.Thisresultsinasuperiorbondedphasewithprovenperformanceandexceptionallifetimeforaphenylreversedphasecolumn.

Properties

YMC-PackPh (Phenyl)providesauniqueselectivitywhencompared toaliphaticstraightchainreversedphasessuchasC18,C8orC4.Theπ-electronsofthephenylgroupscaninteractwitharomaticresiduesofananalytemoleculeinadditiontohydrophobicinteractionstoincreasereten-tionrelativetonon-aromaticspecies.

PhenylphasesareconvenientfortheseparationofaromaticcompoundsandalsoprovideausefulalternativetoC18orC4phasesfortheseparationofpeptidesandproteinsonbothsmallpore(12nm)andwidepore(30nm)materials.Retentionisdecreasedonwideporephenylphasesrelativeto12nmphenylmaterialduetothelowersurfaceareaofthewideporematerial.

YMC-PackPh(Phenyl)isalsoavailableinpreparativeparticlesizes.

Reversed Phase 101



Column: YMC-PackPh(5µm,12nm)250x4.6mmIDPartNo.: PH12S05-2546WTEluent: 100mMtriethylamine-aceticacid(pH6.0)Flow: 1.0ml/minDetection: UVat260nm,0.16AUFSTemperature: 37ºCInjection: 5µl(0.27~0.71mg/ml)

Sugar nucleotides*

1: CDP-D-glucose 2: UDP-D-glucose 3: UDP-N-acetyl-D-glucosamine 4: GDP-D-mannose 5: ADP-D-glucose

Water-soluble vitamins*

Column: YMC-PackPh(5µm,12nm)150x4.6mmIDPartNo.: PH12S05-1546WTEluent: acetonitrile/50mMNH4H2PO4(10/90)Flow: 1.0ml/minDetection: UVat210nm,0.16AUFSTemperature: 37ºCInjection: 10µl(0.02~0.30mg/ml)

1: p-Aminobenzoicacid(PABA) 2: Biotin(VitaminH) 3: Riboflavin(VitaminB2) 4: Cyanocobalamin(VitaminB12)

Column care

YMC-PackPh(Phenyl)isstabletowardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.Ifcolumnsareaffectedbyundesiredcontaminantsorcloggedinletfritswhichcausebackpressureincreases,flushthecolumnwithTHFintheoppositeflowdirection.


Column: YMC-PackPh(5µm,12nm)250x4.6mmIDPartNo.: PH12S05-2546WTEluent: 5mMaceticacidFlow: 1.0ml/minDetection: UVat280nm,0.32AUFSTemperature: 25ºCInjection: 20µl

Polyphenols* Fullerenes*

Column: YMC-PackPh(5µm,12nm)150x6.0mmIDPartNo.: PH12S05-1506WTEluent: hexane/2-propanol(50/50)Flow: 1.0ml/minDetection: UVat350nm,0.08AUFSTemperature: ambient(25ºC)Injection: 4µl(0.125mg/ml)

1

2

3

4

1: Pyrogallol 2: Hydroquinone 3: Catechol 4: Phenol

102030min 1020min

51015min

102 Reversed Phase

General

DuetoshorteralkylchainsYMC-PackC4hasalowerhydrophobicitythanbothC18andC8phases.Thereforeretentiontimesofnon-polarsamplestendtobeshorteronYMC-PackC4,makingitanidealchoiceforfasterseparations.

Properties

YMC-PackC4phasesarelesshydrophobicandgenerallyrequiremoreaqueousbufferthanC8orC18phases.WhencomparedtoC8orC18packingsusingthesameeluent,YMC-PackC4showssignificantlyshorterretentiontimesfornonpolarcompounds.Retentionofpolarcompounds,how-ever,isnotsignificantlyaffected.Therefore,mixtureswithawiderangeofcomponentpolarityarebestseparatedbyYMC-PackC4.Thisisbecausethebutylbondedphasegivesshorterretentiontimeswhilestillmaintaininghighresolutionwhencomparedtolongerchainbondedchemistries.

YMC-Pack C4 Specification


- low hydrophobicity material

- high coverage monomeric bonded chemistry

- ideally suited for separation of biological materials

YMC-Pack C4 (Butyl)

Column: YMC-PackC4(5µm,30nm)150x4.6mmIDEluent: A=water/TFA(100/0.1) B=acetonitrile/2-propanol/TFA(50/50/0.1);30-75%B(0-15min),75%B(15-20min)Flow: 1.0ml/minDetection: UVat220nmTemperature: 37°CInjection: 10µl(0.25~1.0mg/ml)

Proteins (MW 66,000 – 96,000)*

1: BSA (MW66,000) 2: Conalbumin(Chickeneggwhite) (MW77,000) 3: Lipoxidase(Soybean) (MW96,000)

*Impuritiesincommerciallipoxidase

USPL26

Reversed Phase 103


YMC-Pack C4 (Butyl)

Anti-human immunodeficiency virus (HIV) agents*

Column care

YMC-PackC4isstabletowardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.Ifcolumnsareaffectedbyundesiredcontaminantsorcloggedinletfritswhichcausebackpressureincreases,flushthecolumnwithTHFintheoppositeflowdirection.


2,4-Dinitrophenylhydrazones of aldehydes andketones*

Column: YMC-PackC4(5µm,12nm)150x4.6mmIDPartNo.: BU12S05-1546WTEluent: A=tetrahydrofuran/water(10/90) B=acetonitrile;35%B(0-7min),35-65%B(7-18min,linear), 100%B(18-19min),35%B(19-35min)Flow: 1.5ml/minDetection: UVat360nm,0.01AUFSTemperature: 30°CInjection: 11µl(0.0025mg/ml)

Column: YMC-PackC4(5µm,12nm)150x4.6mmIDPartNo.: BU12S05-1546WTEluent: methanol/10mMKH2PO4(10/60)Flow: 1.0ml/minDetection: UVat254nm,0.16AUFSTemperature: 37ºCInjection: 7µl(0.125mg/ml)

1: Formaldehyde2,4-DNPH 2: Acetaldehyde2,4-DNPH 3: Acetone2,4-DNPH 4: Acrolein2,4-DNPH 5: Propionaldehyde2,4-DNPH 6: Crotonaldehyde2,4-DNPH 7: Methylethylketone2,4-DNPH 8: Isobutyraldehyde2,4-DNPH 9: Benzaldehyde2,4-DNPH10: n-Valeraldehyde2,4-DNPH11: p-Tolualdehyde2,4-DNPH12: Capronaldehyde2,4-DNPH

Column: YMC-PackC4(5µm,30nm)150x4.6mmIDPartNo.: BU30S05-1546WTEluent: A)acetonitrile/water/TFA(5/95/0.1) B)acetonitrile/water/TFA(60/40/0.1)Flow: 1.0ml/minDetection: UVat220nm,0.32AUFSTemperature: 37ºCInjection: 16µl(0.16~0.33mg/ml)

Proteins* Water-soluble vitamins*

Column: YMC-PackC4(5µm,12nm)150x4.6mmIDPartNo.: BU12S05-1546WTEluent: acetonitrile/50mMNH4H2PO4(10/90)Flow: 1.0ml/minDetection: UVat210nm,0.16AUFSTemperature: 37ºCInjection: 10µl(0.02~0.30mg/ml)

1: RibonucleaseA 2: Cytochromec 3: Lysozyme 4: Myoglobin

1: p-Aminobenzoicacid(PABA) 2: Biotin(VitaminH) 3: Riboflavin(VitaminB2) 4: Cyanocobalamin(VitaminB12)

510152025min

1020min

1020min1020min

104 Reversed Phase

General

YMC-PackC1(TMS)isabondedphasesuitableforsamplesthatexhibitstrongretentionchar-acteristicsandaredifficultorimpossibletoseparateonconventionalreversedphaseornormalphasecolumns.

Properties

YMC-PackTMS(C1)isbondedwithtrimethylmonochlorosilanetocreateaphasewithintermediatepolarityforseparationofextremelyhydrophobiccompoundsusingconventionalreversedphasesolventsandofhighlypolarcompoundsusingnormalphasesolvents.

ThechemistryofYMC-PackTMS(C1)isalsowell-suitedfortheanalysisofmultifunctionalcom-pounds.SelectivitycharacteristicsofaC1bondedphasecanbeunique,andsamplesmustbetestedtodeterminethesuitabilityofthephase.

YMC-PackTMS(C1)isalsoavailableinpreparativeparticlesizes.

- stationary phase with the lowest hydrophobicity among reversed phase packing materials

- intermediate polarity between normal phase silica and other alkyl bonded reversed phases

- for fast separations of highly hydrophobic compounds

- alternative to C18 for the separation of hydrophilic compounds

YMC-Pack TMS (C1)

YMC-Pack TMS Specification


USPL13

Reversed Phase 105


YMC-Pack TMS (C1)

Water-soluble vitamins* Water-soluble vitamins*

1: L-Ascorbicacid (VitaminC)

2: Oroticacid (VitaminB13)

3: Pyrroloquinolinequinone (PQQ)

4: PyridoxineHCl (VitaminB6)

5: Nicotinamide

1: p-Aminobenzoicacid (PABA)

2: Biotin (VitaminH)

3: Riboflavin (VitaminB2)

4: Cyanocobalamin (VitaminB12)

Column care

YMC-PackTMS(C1)isstabletowardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.Storethecolumninmethanol/water=70/30.Ifcolumnsareaffectedbyundesiredcontaminantsorcloggedinletfritswhichcausebackpressureincreases,flushthecolumnwithTHFintheoppositeflowdirection.


510min 1020min

Column: YMC-PackTMS(C1)(5µm,12nm)150x4.6mmIDPartNo.: TM12S05-1546WTEluent: 100mMCH3COOH/100mMCH3COONH4(30/70,pH5.1)Flow: 1.0ml/minDetection: UVat254nm,0.16AUFSTemperature: 37°CInjection: 10µl(0.04~0.20mg/ml)

Column: YMC-PackTMS(C1)(5µm,12nm)150x4.6mmIDPartNo.: TM12S05-1546WTEluent: acetonitrile/50mMNH4H2PO4(10/90)Flow: 1.0ml/minDetection: UVat210nm,0.16AUFSTemperature: 37ºCInjection: 10µl(0.02~0.30mg/ml)

106 Reversed Phase

YMC-Pack CN (Cyano)

General

Inreversedphasemode,cyano(nitrile)phasesarethemostpolarandleastretentiveofallreversedphasesupports.Extremelyhydrophobiccompounds,whichdonoteluteonstandardC18andC8columnswithtypicalreversedphaseeluents,canbeseparatedusingcyanophases.SeparationsusingreversedandnormalphaseandHILICmechanismscanbecarriedoutusingthismaterial.

Properties

ThecyanochemistryofYMC-PackCN(Cyano)providesadifferentselectivityfrombothphenylandstandardaliphatic(C18,C8orC4)reversedphases.Itisusefulforquickandsimpleanalysisofcompoundsthatdiffergreatlyinhydrophobicity,withouttheneedtousegradientelutionchro-matography.

Cyanopackingsalsoprovideanalternativetosilicamaterialinnormalphasechromatography,wherebondednormalphasepackingshavetheadvantageoffasterequilibration,moreuniformsurfaceactivityandincreasedresistancetodissolution.

Toextendcolumnlifetimecontinuedswitchingbetweennormalandreversedphasesolventsshouldbeavoided.

YMC-PackCN(Cyano)isalsoavailableinpreparativeparticlesizes.

- for normal, reversed phase and HILIC applications

- silica gel with cyanopropyl groups

- faster column equilibration than normal silica gel

- most polar reversed phase column

YMC-Pack CN Specification


USPL10

Reversed Phase 107


YMC-Pack CN (Cyano)

Column: YMC-PackCN(Cyano)(5µm,12nm)250x4.6mmIDPartNo.: CN12S05-2546WTEluent: acetonitrile/water/TFA(15/85/0.05)Flow: 0.5ml/minDetection: UVat220nm,0.64AUFSTemperature: 30ºCInjection: 5µl

Aspartame in a soft drink*

Column care

YMC-PackCN(Cyano)isstabletowardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.Fordetailedinformationpleaserefertothe“ColumnCareandUseInstructions”whichareshippedwitheachanalyticalcolumn.

Column: YMC-PackCN(Cyano)(5µm,12nm)250x4.6mmIDPartNo.: CN12S05-2546WTEluent: acetonitrile/water(40/60)Flow: 0.6ml/minDetection: UVat254nm,0.32AUFSTemperature: 30°CInjection: 3µl(0.25mg/ml)

Steroids*

1. Caffeine 2. Aspartama 3. Benzoicacid

1020min

108 Reversed Phase


Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 AQ12S03-H3Q1QT AQ12S03-05Q1QT AQ12S03-10Q1QT AQ12S03-15Q1QT AQ12S03-25Q1QT AQ12S03-01Q1GC3.0 AQ12S03-H303QT AQ12S03-0503QT AQ12S03-1003QT AQ12S03-1503QT AQ12S03-2503QT AQ12S03-0103GC4.0 AQ12S03-H304QT AQ12S03-0504QT AQ12S03-1004QT AQ12S03-1504QT AQ12S03-2504QT AQ12S03-0104GC4.6 AQ12S03-0346WT AQ12S03-0546WT AQ12S03-1046WT AQ12S03-1546WT AQ12S03-2546WT AQ12S03-0104GC

20 nm3 µm


12 nm5 µm


20 nm5 µm


YMC-Pack ODS-AQ

Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 AA12S03-H3Q1QT AA12S03-05Q1QT AA12S03-10Q1QT AA12S03-15Q1QT AA12S03-25Q1QT AA12S03-01Q1GC3.0 AA12S03-H303QT AA12S03-0503QT AA12S03-1003QT AA12S03-1503QT AA12S03-2503QT AA12S03-0103GC4.0 AA12S03-H304QT AA12S03-0504QT AA12S03-1004QT AA12S03-1504QT AA12S03-2504QT AA12S03-0104GC4.6 AA12S03-0346WT AA12S03-0546WT AA12S03-1046WT AA12S03-1546WT AA12S03-2546WT AA12S03-0104GC

20 nm3 µm


12 nm5 µm


20 nm5 µm


30 nm5 µm


YMC-Pack ODS-A

Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 AM12S03-H3Q1QT AM12S03-05Q1QT AM12S03-10Q1QT AM12S03-15Q1QT AM12S03-25Q1QT AM12S03-01Q1GC3.0 AM12S03-H303QT AM12S03-0503QT AM12S03-1003QT AM12S03-1503QT AM12S03-2503QT AM12S03-0103GC4.0 AM12S03-H304QT AM12S03-0504QT AM12S03-1004QT AM12S03-1504QT AM12S03-2504QT AM12S03-0104GC4.6 AM12S03-0346WT AM12S03-0546WT AM12S03-1046WT AM12S03-1546WT AM12S03-2546WT AM12S03-0104GC

12 nm5 µm

2.1 AM12S05-H3Q1QT AM12S05-05Q1QT AM12S05-10Q1QT AM12S05-15Q1QT AM12S05-25Q1QT AM12S05-01Q1GC3.0 AM12S05-H303QT AM12S05-0503QT AM12S05-1003QT AM12S05-1503QT AM12S05-2503QT AM12S05-0103GC4.0 AM12S05-H304QT AM12S05-0504QT AM12S05-1004QT AM12S05-1504QT AM12S05-2504QT AM12S05-0104GC4.6 AM12S05-0346WT AM12S05-0546WT AM12S05-1046WT AM12S05-1546WT AM12S05-2546WT AM12S05-0104GC

YMC-Pack ODS-AM




Reversed Phase 109



Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 AL12S03-H3Q1QT AL12S03-05Q1QT AL12S03-10Q1QT AL12S03-15Q1QT AL12S03-25Q1QT AL12S03-01Q1GC3.0 AL12S03-H303QT AL12S03-0503QT AL12S03-1003QT AL12S03-1503QT AL12S03-2503QT AL12S03-0103GC4.0 AL12S03-H304QT AL12S03-0504QT AL12S03-1004QT AL12S03-1504QT AL12S03-2504QT AL12S03-0104GC4.6 AL12S03-0346WT AL12S03-0546WT AL12S03-1046WT AL12S03-1546WT AL12S03-2546WT AL12S03-0104GC

12 nm5 µm

2.1 AL12S05-H3Q1QT AL12S05-05Q1QT AL12S05-10Q1QT AL12S05-15Q1QT AL12S05-25Q1QT AL12S05-01Q1GC3.0 AL12S05-H303QT AL12S05-0503QT AL12S05-1003QT AL12S05-1503QT AL12S05-2503QT AL12S05-0103GC4.0 AL12S05-H304QT AL12S05-0504QT AL12S05-1004QT AL12S05-1504QT AL12S05-2504QT AL12S05-0104GC4.6 AL12S05-0346WT AL12S05-0546WT AL12S05-1046WT AL12S05-1546WT AL12S05-2546WT AL12S05-0104GC

YMC-Pack ODS-AL

Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

6 µm

2.1 PC99S06-H3Q1QT PC99S06-05Q1QT PC99S06-10Q1QT PC99S06-15Q1QT PC99S06-25Q1QT PC99S06-01Q1GC3.0 PC99S06-H303QT PC99S06-0503QT PC99S06-1003QT PC99S06-1503QT PC99S06-2503QT PC99S06-0103GC4.0 PC99S06-H304QT PC99S06-0504QT PC99S06-1004QT PC99S06-1504QT PC99S06-2504QT PC99S06-0104GC4.6 PC99S06-0346WT PC99S06-0546WT PC99S06-1046WT PC99S06-1546WT PC99S06-2546WT PC99S06-0104GC

YMC-Pack PolymerC18

Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 OC12S03-H3Q1QT OC12S03-05Q1QT OC12S03-10Q1QT OC12S03-15Q1QT OC12S03-25Q1QT OC12S03-01Q1GC3.0 OC12S03-H303QT OC12S03-0503QT OC12S03-1003QT OC12S03-1503QT OC12S03-2503QT OC12S03-0103GC4.0 OC12S03-H304QT OC12S03-0504QT OC12S03-1004QT OC12S03-1504QT OC12S03-2504QT OC12S03-0104GC4.6 OC12S03-0346WT OC12S03-0546WT OC12S03-1046WT OC12S03-1546WT OC12S03-2546WT OC12S03-0104GC

20 nm3 µm


12 nm5 µm


20 nm5 µm


30 nm5 µm


YMC-Pack C8

Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 PH12S03-H3Q1QT PH12S03-05Q1QT PH12S03-10Q1QT PH12S03-15Q1QT PH12S03-25Q1QT PH12S03-01Q1GC3.0 PH12S03-H303QT PH12S03-0503QT PH12S03-1003QT PH12S03-1503QT PH12S03-2503QT PH12S03-0103GC4.0 PH12S03-H304QT PH12S03-0504QT PH12S03-1004QT PH12S03-1504QT PH12S03-2504QT PH12S03-0104GC4.6 PH12S03-0346WT PH12S03-0546WT PH12S03-1046WT PH12S03-1546WT PH12S03-2546WT PH12S03-0104GC

12 nm5 µm


30 nm5 µm







110 Reversed Phase


Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 TM12S03-H3Q1QT TM12S03-05Q1QT TM12S03-10Q1QT TM12S03-15Q1QT TM12S03-25Q1QT TM12S03-01Q1GC3.0 TM12S03-H303QT TM12S03-0503QT TM12S03-1003QT TM12S03-1503QT TM12S03-2503QT TM12S03-0103GC4.0 TM12S03-H304QT TM12S03-0504QT TM12S03-1004QT TM12S03-1504QT TM12S03-2504QT TM12S03-0104GC4.6 TM12S03-0346WT TM12S03-0546WT TM12S03-1046WT TM12S03-1546WT TM12S03-2546WT TM12S03-0104GC

12 nm5 µm


30 nm5 µm


YMC-Pack TMS

Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

12 nm3 µm

2.1 CN12S03-H3Q1QT CN12S03-05Q1QT CN12S03-10Q1QT CN12S03-15Q1QT CN12S03-25Q1QT CN12S03-01Q1GC3.0 CN12S03-H303QT CN12S03-0503QT CN12S03-1003QT CN12S03-1503QT CN12S03-2503QT CN12S03-0103GC4.0 CN12S03-H304QT CN12S03-0504QT CN12S03-1004QT CN12S03-1504QT CN12S03-2504QT CN12S03-0104GC4.6 CN12S03-0346WT CN12S03-0546WT CN12S03-1046WT CN12S03-1546WT CN12S03-2546WT CN12S03-0104GC

12 nm5 µm


30 nm5 µm


YMC-Pack CN (Cyano)

Phase dimension

Column ID



[pack of 5]

30 (WT) / 33 (QT) 50 100 150 250

3 µm

2.1 BA99S03-H3Q1QT BA99S03-05Q1QT BA99S03-10Q1QT BA99S03-15Q1QT BA99S03-25Q1QT BA99S03-01Q1GC3.0 BA99S03-H303QT BA99S03-0503QT BA99S03-1003QT BA99S03-1503QT BA99S03-2503QT BA99S03-0103GC4.0 BA99S03-H304QT BA99S03-0504QT BA99S03-1004QT BA99S03-1504QT BA99S03-2504QT BA99S03-0104GC4.6 BA99S03-0346WT BA99S03-0546WT BA99S03-1046WT BA99S03-1546WT BA99S03-2546WT BA99S03-0104GC

5 µm

2.1 BA99S05-H3Q1QT BA99S05-05Q1QT BA99S05-10Q1QT BA99S05-15Q1QT BA99S05-25Q1QT BA99S05-01Q1GC3.0 BA99S05-H303QT BA99S05-0503QT BA99S05-1003QT BA99S05-1503QT BA99S05-2503QT BA99S05-0103GC4.0 BA99S05-H304QT BA99S05-0504QT BA99S05-1004QT BA99S05-1504QT BA99S05-2504QT BA99S05-0104GC4.6 BA99S05-0346WT BA99S05-0546WT BA99S05-1046WT BA99S05-1546WT BA99S05-2546WT BA99S05-0104GC

YMCbasic

For other dimensions please refer to page 247




112 Normal Phases

Normal Phases 113


Contents

•YMC-PackSIL(Silica) ...................................... 114-115

•YMC-PackPVA-Sil ........................................... 116-117

•YMC-PackCN(Cyano) .................................... 118-119

•YMC-PackDiol-NP .......................................... 120-121

•YMC-PackPolyamineII .................................. 122-123

•YMC-PackNH2(Amino) ..........................................124

•YMC-PackTMS(C1) ...............................................125

•OrderingInformation ........................................ 126-127

Normal Phase Chemistries

HPLC Columns for Normal Phase ChromatographyWhilsthistoricallyitwastheearliestformofHPLC,normal-phaseseparationshaverecentlylessattentionduetothebeliefthatitiscomplicatedandunpredictable.Butnormal-phasechromatographyisapowerfultoolfortheseparationofpositionalisomersthataredifficulttoseparateinreversed-phasemode.Duetoarigidsurfaceincomparisonwiththemoreflexiblecarbonchainsofreversed-phasestationaryphasestheanalytesareeffectedbywelldefinedstericinteractionwithpolargroups.

ThissectiongivesacomprehensiveoverviewofthestationaryphasesavailablefromYMCfortheuseinnormalphasesepa-rationmode.YMCofferscolumnspackedwithnon-bondedsilicaorpackedwithsilicagelmodifiedwithpolargroups.

Introduction

114 Normal Phases

YMC-Pack SIL (Silica)

General

DuetothehighlysophisticatedproductionprocessYMC’ssphericalsilicamaterialshowsoutstand-ingperformanceandgreatlot-to-lotreproducibility.Thereasonforthiscanbesummarisedintwomainqualities:verynarrowphysicalandchemicalproductspecificationsandoutstandingpurity.

Properties

The high purity YMC-PackSIL (Silica) allowsalmost totalsample recoverybecause the lowcontent of impurities such as residualmetalsreducesnon-specific sample adsorption. Thisalsopreventsunusualpeak-shapestherebyen-couraging higher sample loading. In addition,theporousstructureofthespheresgivesahighsurface areawhich further improves sampleloading.

Comparedwith irregular silica,YMC’s spheri-calmaterial issubjecttoamuchlowerdegreeofmechanicaldegradationduringpackingandusage.Thisresultsinlowerbackpressuresandextendedcolumnlifetimesduetotheabsenceof‘fines’.

SinceYMCsphericalsilicaisthebasisforeveryYMCbondedphase,thisisafurtherreasonforthepremiumqualityofYMCstationaryphasesas far as backpressure and chromatographicstability is concerned.

YMC-PackSIL(Silica)isalsoavailableinprepara-tiveparticlesizes.

- ultra high purity silica

- high mechanical stability

- highly porous, totally spherical particles

- fully scalable for analytical, semi-prep, preparative and process scale applications

- convenient for separating small organic compounds with similar structures

Ultra High Purity Silica*

1. a-Tocopherol 2. b-Tocopherol 3. g-Tocopherol 4. d-Tocopherol

YMC-Pack SIL Specification

Particle Size / µm 3; 5 3; 5 3; 5 5Pore Size / nm 6 12 20 30Surface area / m2g-1 450 330 175 100Recommended pH range 2.0 - 7.5 2.0 - 7.5 2.0 - 7.5 2.0 - 7.5

Column: YMC-PackSIL(5µm,12nm)250x4.6mmIDPartNo.: SL12S05-2546WTEluent: hexane/2-propanol/aceticacid(1000/6/5)Flow: 1.4ml/minDetection: FLSatEx298nm,Em325nmTemperature:35ºCInjection: 20µl(5~20mg/ml)

Normal Phases 115


YMC-Pack SIL (Silica)

Column: YMC-PackSIL(5µm,6nm)250x4.6mmIDPartNo.: SL06S05-2546WTEluent: hexane/THF/aceticacid(60/40/1)Flow: 1.5ml/minDetection: UVat250nm,0.16AUFSTemperature: 30°CInjection: 10µl(0.25mg/ml)

Tocopherols*

Column: YMC-PackSIL(5µm,12nm)150x4.6mmIDPartNo.: SL12S05-1546WTEluent: hexane/THF/aceticacid(97/3/0.25)Flow: 1.0ml/minDetection: UVat295nm,0.08AUFSTemperature: 30ºCInjection: 10µl(0.20~0.31mg/ml)

1:a-Tocopherol 2: b-Tocopherol 3: g-Tocopherol 4: d-Tocopherol

1: Retinolpalmitate (VitaminApalmitate)

Column care

YMC-PackSIL isstable towardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.Fordetailedinformationpleaserefertothe“ColumnCareandUseInstructions”whichareshippedwitheachanalyticalcolumn.

5min

102030min

10min

Steroids*

Vitamin A fatty acid ester*

Column: YMC-PackSIL(5µm,12nm)150x4.6mmIDPartNo.: SL12S05-1546WTEluent: cyclohexane/dichloromethane/aceticacid(100/0.5/0.5)Flow: 1.0ml/minDetection: UVat350nm,0.32AUFSTemperature: 30ºCInjection: 5µl(1.0mg/ml)

116 Normal Phases

Polyvinyl Alcohol Functionalised Silica

PVA-Silispreparedfroma5micron12nmsilicasupportwhichisbondedwithamonomolecularpolymercoatingofvinylalcohol.ThepolymerisedPVAcompletelycoversbothexternalandinternalsurfacesofthesilicasupport,protectingitagainstaggressive,highpHbuffersandsolvents.

Normal phase alternative to Silica

PVA-Sil,whichpossessesapolyvinylalcohol(PVA)surfacechemistry,isanexcellentalternativetosilicagelorotherpolarbondedphaseswhichareusedinnormalphasechromatography.Inmanysituationsitexhibitsbetterperformancecharacteristicsandauniqueselectivityandcanoftenresolvecompoundsthatbehavepoorlyonsilica.ThealcoholfunctionalitypresentonPVA-Silisbettersuitedfortroublesomecompounds,suchorganicbases,thanacidicsilanolspresentinunbondedsilica.

Highly stable and reproducible

SincePVA-Silisabondedstationaryphase,itcanbewashedwithsolventsofanypolarity,fromhexanethroughwater,withoutalteringthesurfaceactivity.Thereforeselectivity,retentionandresolu-tionarereproducibleregardlessofthecolumn’sprevioushistory.Thisisnottrueofbaresilica,whicheasilybecomescompletelydeactivatedfollowingtheintroductionofevensmallquantityofwater.

Provides high sample recovery

ThesurfaceofPVA-Silisveryuniformwithoutthehighlyactiveacidicsilanolsitesonbaresilicawhichcancausedecompositionofsensitivemolecules.Becauseofconsistentsurfaceactivity,PVA-Sil exhibits neither non-specific irreversible adsorption nor sampledegradation. This is aproblemoftenencounteredwithbaresilicacolumns.Thelackofnon-specificadsorptionandtheuniformityofthepolyvinylalcoholbondedsurfacemeansthat,unlikesilica,PVA-Silcanbereusedoverandoverwithoutfearofcontaminationorcarryover.SamplerecoveriesonPVA-Siltypicallyaverage90%orhigher.

Excellent choice for packed column SFC

ThePVApolymershellonPVA-Sildeactivatesthesilicasupportwhileprovidingahydrophilicsur-face.This,coupledwithavailablecolumndimensionof1.0mmand2.0mmIDmeansthatPVA-SilcolumnsarewellsuitedforSFCseparations.

Column Care

YMC-PackPVA-SilisstabletowardshydrolysisbetweenpH2.0-9.5.Removeacidandbuffersaltsbeforestorage.Fordetailedinformationpleaserefertothe“ColumnCareandUseInstructions”whichareshippedwitheachanalyticalcolumn.

- bonded phase alternative to silica for normal phase applications

- vinyl alcohol polymerised silica support

- consistent surface activity, unaffected by water

- excellent for packed column supercritical fluid chromatography

YMC-Pack PVA-Sil

USPL24

YMC-Pack PVA-Sil Specification

Particle Size / µm 5Pore Size / nm 12Surface area / m2g-1 330Recommended pH range 2.0 - 9.5

Normal Phases 117


YMC-Pack PVA-Sil

Uracil (in HILIC-mode)*

Column: YMC-PackPVA-Sil(5µm,12nm)100x3.0mmIDPartNo.: PV12S05-1003WTEluent: acetonitrile/CH3COONH4;200mM,pH5,5 isocratic(95/5)Flowrate: 0.9ml/minDetection: UVat275nm

high stability

Analysis of Potato Lipids*

Column: YMC-PackPVA-Sil(5µm,12nm)250x4.6mmIDPartNo.: PV12S05-2546WTFlowrate: 1to2ml/minMobilePhase: A: iso-hexane/methylterbutylether(98:2) B: propan-2-ol/ACN/CHCl3/CH3OOH(84:8:8:0.025) C: propan-2-ol/water/triethylamine(50:50:0.2)Gradient:Tmin: 0 5 15 40 40.1 45 50B%: 0 20 52 52 70 0 0C%: 0 0 4 14 0 0 0Flow(ml/mn): 1 1 1 1.4 1.4 2 2

Nebulisertemperature:25°C,Evaporationtemperature:35°C

S: Sterols SE: SterolEsters SG: SterylglycosidesMGDG:Monogalactosyldiacylglycerols DGDG: DigalactosyldiacylglycerolsPE: Phosphatidylethanolamine PG: PhosphatidylglycerolsPC: Phosphatidylcholine ASG: AcylsterylglycosidesPI: Phosphatidylinositol DG: DiacylglycerolCER: Cerebrosides

Column: YMC-PackPVA-Sil(5µm,12nm)250x4.0mmIDPartNo.: PV12S05-2504QTEluent: A:n-hexane/tert-methylbutylether(98:2) B:isopropanol/acetonitrile/chloroform/aceticacid(84:8:8:0.025) C:isopropanol/water/triethylamine(50:50:0.2) plus5mMammoniumsulfateFlowrate: 1ml/minDetector: ELSD

SE: steryloleateTG: TAG/tripentadecanoinSterol: stigmasterol/sitosterolSQDG: sulfoquinovosyldiacylglycerolDHCer: dehydroxycerebrosideCer: cerebrosideDGDG: digalactosyldiacylglycerol

PE: PE-dipalmitoylPA: PA-diheptadecanoylPI: PI-diheptadecanoylPS: PS-diheptadecanoylPC: PC-diheptadecanoylSPH: sphingomyelinLPC: lysophosphatidylcholine

Literature:JAOCS,Vol.80,no.8(2003)p.747-753

Literature:W.W.Christie;R.A.Urwin,J.highResol.Chromatogr.,Vol.18(1995)p.97-100

Analysis of Lipids

118 Normal Phases

YMC-Pack CN (Cyano)

General

Cyanopackingsalsoprovideanalternativetosilicamaterialinnormalphasechromatography,wherebondednormalphasepackingshavetheadvantageoffasterequilibration,moreuniformsurfaceactivityandincreasedresistancetodissolution.

Toextendcolumnlifetimecontinuedswitchingbetweennormalandreversedphasesolventsshouldbeavoided.Bothreversedandnormalphaseseparationscanbecarriedoutonthismaterial.

YMC-PackCN(Cyano)isalsoavailableinpreparativeparticlesizes.

- silica gel chemically bound with cyanopropyl groups

- faster column equilibration than normal silica gel

USPL10

YMC-Pack CN Specification

Particle Size / µm 3; 5 5Pore Size / nm 12 30Surface area / m2g-1 330 175Carbon content / % 7 2.5Recommended pH range 2.0 - 7.5 2.0 - 7.5

Normal Phases 119


YMC-Pack CN (Cyano)

Column: YMC-PackCN(5µm,12nm)250x4.6mmIDPartNo.: CN12S05-2546WTEluent: hexane/methanol/dichloromethane(75/20/5)Flow: 0.6ml/minDetection: UVat254nm,0.32AUFSTemperature: 30ºCInjection: 3µl(0.25mg/ml)

Steroids in normal phase*

Column care

YMC-PackCNisstabletowardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.Fordetailedinformationpleaserefertothe“ColumnCareandUseInstructions”whichareshippedwitheachanalyticalcolumn.

Column: YMC-PackCN(5µm,12nm)250x4.6mmIDPartNo.: CN12S05-2546WTEluent: acetonitrile/water(40/60)Flow: 0.6ml/minDetection: UVat254nm,0.32AUFSTemperature: 30°CInjection: 3µl(0.25mg/ml)

Steroids in reversed phase*

YMC-Pack CN Separation Modes

YMC-PackCNcanbeusedeitherinreversed-phaseandnormal-phasemodessinceitprovidescyanopropylgroupsofmediumpolarity.Itcanbeemployedinreversed-phasemodewithanaqueousmobilephaseofhigherpolarityandinnormal-phasemodewithalowerpolaritythanthestationaryphase.Thisresultsinanimportantphenomenonforlarge-scalework;theelutionorderwillbeinvertedbyuseofthealternateseparationmode.

1020min 1020min

120 Normal Phases

YMC-Pack Diol-NP

General

InnormalphasemodetheYMC-PackDiolstationaryphaseisaversatilealternativetosilica.Thebondedphase’shydroxylgroupsprovidegoodselectivitywithoutexcessiveretention,sincehy-drogenbondingwiththediollayerisnotasstrongaswiththesilanolsonabaresilicasurface.Diolcolumnsalsoprovideimprovedreproducibilitywhencomparedwithbaresilica.

Diolpackingsaresuitableforseparationsusingreversedphasetechniquesormolecularweightdeterminationofproteinsbygelfiltration.

Properties

AswithallYMCsilicabasedbondedphases,YMC-PackDiolstartswithabasesilicasupportofexceptionalpurity.YMCmanufacturingandqualitycontrolproceduresensurethatthesilicahasaverylowresidualmetalcontent.Thesilicapuritygreatlyreducesnon-specificsampleadsorption,therebyprovidingexcellentsamplerecovery.

Thehighsurfacearea,togetherwiththelargenumberofavailablesitesforinteractionofthe1,2-dihydroxypropaneligands,provideshighpreparativeloading.

YMC-PackDiolGPCcolumnsexhibitbetterperformancecharacteristicsthanunderivatisedsilicaforsizeseparationsasthenon-specificadsorptivesiteshavebeeneliminated.Diolisavailableinfourporosities:6,12,20and30nmandthusitissuitableforseparationormolecularweightdeter-minationofproteinswithmolecularweightsof10,000toseveralhundredthousands.

YMC-PackDiolpackingscanbecleanedrepeatedlywithmethanol,orevenwater.Whencombinedwiththehighmechanicalstrengthofthepurebasesilica,thiswashabilitymeansthatYMC*GelDiolpackingsprovidelongercolumnlifethanunderivatisedsilica.

YMC-PackDiolisalsoavailableinpreparativeparticlesizes.

- good selectivity without excessive retention

- high product recovery rate

- high prep loading

- bonded phase reproducibility

- improved peak shape versus bare silica

- gel filtration on a silica based material for aqueous size separations

USPL20

YMC-Pack Diol-NP Specification

Particle Size / µm 5 5Pore Size / nm 6 12Surface area / m2g-1 450 330Recommended pH range (DN)

(DL)2.0 - 7.55.0 - 7.5

2.0 - 7.55.0 - 7.5

Normal Phases 121


YMC-Pack Diol-NP

Separations of phenols*

Column: YMC-PackDiol-NP(DN)(5µm,12nm)250x4.6mmIDPartNo.: DN12S05-2546WTEluent: hexane/ethanol(80/20)Flowrate: 1.0ml/minTemperature: 30°CDetection: UVat254nm

1. Phenol 2. Catechol 3. Resorcinol 4. Hydroquinone 5. Pyrogallol 6. Phloroglucinol

Column care

YMC-PackDiolisstabletowardshydrolysisbetweenpH5.0-7.5inreversedphasemode(DL)andpH2.0-7.5innormalphasemode(DN).Removeacidandbuffersaltsbeforestorage.Fordetailedinformationpleaserefertothe“ColumnCareandUseInstructions”whichareshippedwitheachanalyticalcolumn.

122 Normal Phases

General

Thechromatographicseparationandthereliablequantitationofsaccharidesisincreasinglyimportantinmanyareasoffoodtechnology,lifescienceandinpharmaceuticalindustry.

Fortheseparticularapplications,YMCprovidesYMC-PackPolyamineII,apolymeraminophase.

Properties

YMC-PackPolyamineIIisbasedonultra-pureYMCsilicaasasupportmaterial.Thefunctionalityofthestationaryphaseisachievedbyacovalentlybondedpolymerlayercontainingsecondary(2°)andtertiary(3°)aminogroups.The2°and3°aminogroupsofYMC-PackPolyamineIIareonlyweaklynucleophilic,exhibitingasignificantlyreducedreactivityagainstcarbonylcompounds.Therefore,unlikeconventionalaminophaseswithprimaryn-propyl-aminoligands,YMC-PackPolyamineIIdoesnottendtotheformationofSchiff’sbasesorotherstablecondensationproducts.Inaddition,the2°and3°aminogroupsofthepolymerlayeraretoalargeextentresistanttooxidationandhydrolysis(seefigurenextpage).

Thelowreactivityofthe2°and3°aminogroupspreservesthelong-termretentioncharacteristicsandselectivityofYMC-PackPolyamineII.

Comparedtoconventionalaminophases,oneoftheirmostoutstandingbenefitsisthesignificantlyprolongedlifetime.Asthesilicamatrixiscompletelypolymercoated,eventheshort-termuseofbasiceluentsuptopH10.5ispossible.

Reducingsugarsareoftenadsorbedirreversiblytoconventionalaminophases,whichcausesprob-lemsintheirrecoveryandquantitation.InYMC-PackPolyamineIIcolumnshowever,theadsorptionofreducingsugarsplaysonlyaminorrole.Asaresultahighrecoveryofthesecompoundscanbeobtainedwhichisbeneficialforaccurateandreliablequantitation.

Column care

YMC-PackPolyamineIIisstabletowardshydrolysisbetweenpH2.0-9.0.Removeacidandbuffersaltsbeforestorage.Fordetailedinformationpleaserefertothe“ColumnCareandUseInstructions”whichareshippedwitheachanalyticalcolumn.

- amino phase with polymeric surface- exclusively 2° and 3° amino groups- stable towards hydrolysis and oxidation- high recovery- excellent life-time

- saccharides and derivatives- nucleotides- tocopherols- for RP- and NP-mode separations

YMC-Pack Polyamine II

YMC-Pack Polyamine II Specification

Particle Size / µm 5Pore Size / nm 12Surface area / m2g-1 n/aCarbon content / % n/aRecommended pH range 2.0 - 7.5

Normal Phases 123


YMC-Pack Polyamine II

Tocopherols*

Column: YMC-PackPolyamineII(5µm,12nm)250x4.6mmIDPartNo.: PB12S05-2546WTEluent: hexane/ethylacetate=70/30Flow: 1.0ml/minDetection: UV,295nmTemperature: 30ºC

1

a-Tocopherol

3

g-Tocopherol

b-Tocopherol

2

d-Tocopherol

4

Ret

enti

on

tim

e

Ret

enti

on

tim

e

[a] conventional amino phase [b] YMC-Pack Polyamine II

continuous flow of distilled water /h

Stability of amino type packings*

10203040min

124 Normal Phases

General

YMC-PackNH2(Amino)packingsarespecificallyusefulfortheanalysisofmono-andpolysac-charidesunderaggressivenormalphaseelutionconditions.Theycanalsobeusedinplaceofsilicaforconventionalnormalphasechromatographyusingnonpolarsolvents.

Properties

YMC-PackNH2(Amino)isbasedonamonomericbondingofaprimarypropylaminefunctionalitytoYMC’sspherical,ultrapure,highsurfaceareasilicawithameanporediameterof12nm.Theaminefunctionalityprovidesretentionandallowstheseparationofpolarcompoundsunderaggressivenormalphaseelutionconditions,e.g.theanalysisofmono-andpolysaccharidesusingacetonitrile/watereluents.(SinceYMC-PackNH2packingsoperateundernormalphase/HILICelutioncondi-tions,water,whichismorepolarthanacetonitrile,isthestrongersolvent.)YMC-PackNH2(Amino)canalsobeusedfortheseparationofisomersoftocopherolsandotherorganicsolublecompoundssuchasparaffins,olefinsandaromaticsunderconventionalnormalphaseconditions.

Inaqueous,lowpHbufferstheaminophasebecomesaweakanionexchangercapableofseparat-ingnegativelychargedmolecules.

YMC-PackNH2(Amino)isalsoavailableinpreparativeparticlesizes.

Column care

YMC-PackNH2(Amino)isstabletowardshydrolysisbetweenpH2.0-7.5.Removeacidandbuffersaltsbeforestorage.


- primary amine (-NH2) functionality

- stable, high coverage monomeric bonded chemistry

- available in analytical, semi-prep and preparative column sizes

YMC-Pack NH2 (Amino)

USPL8

YMC-Pack NH2 Specification

Particle Size / µm 3; 5Pore Size / nm 12Surface area / m2g-1 330Recommended pH range 2.0 - 7.5

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