uv vis spectroscopy
DESCRIPTION
Power point Presentation for working of Ultraviolet Vis spectroscopyTRANSCRIPT
Buck Scientific, Inc.
Seminar in ANALYTICAL INSTRUMENTATION for
Spectroscopy & Chromatography:
UV-Vis Spectrophotometry
Ultra-Violet/Visible SPECTROSCOPY (UV-Vis)
A TYPE of VIBRATIONAL Spectroscopy, similar to Infra-Red. A Sample in a Cuvette is
exposed to Light Energy between 190nm. (in the UV) and 1000nm. (in the VIS). Certain parts of an Organic Molecule will ABSORB some of this energy to create PEAKS on a SPECTRUM for
Quantitative (primarily) and Qualitative Analysis
Theory & Principle of OPERATION
BASIC Spectrophotometer Design
Like AAS instrumentation, the ORIGINAL UV-Vis Specs were made as DOUBLE-BEAM units to correct for Noise, Drift and other Instabilities. Over 20 years ago, the well-known leaders in the Analytical Instrument markets; Beckman & Perkin-Elmer; began to focus on a line of “Stable-Beam” SINGLE-BEAM Instruments.
The BUCK/Cecil series employs this optimum design in our instruments with unique Internal software
Basic SPECTROMETER Design
ALL Analytical Spectrometers have several BASIC design components in common:
Light Source
Sample “Chamber”
Optical System
Detector
Output
Basic SPECTROMETER Design
ALL Analytical Spectrometers have several BASIC design components in common:
LIGHT SOURCE: External D2 & W Lamps to prevent thermal instability within instrument (no fan)
SAMPLE “Chamber”: Standard & Specialty Holders
OPTICS: High-energy Ebert Monochromator DETECTOR: Choice of low-Noise Photo-Diode or
high-Sensitivity Photo-Multiplier Tube
OUTPUT: Graphics On-Screen Display, Analog, Printer and Serial RS-232 P-C capabilities
UV/Vis Important Features
• Scan Speed
• Resolution
• Software Features
• Ease of Operation
• Data Storage
• Customized Calculations
• Support & Service
BUCK/Cecil Instruments
The COMPACT, “Stable-Beam” construction of the Single-Beam series uses the high-quality Cecil base instrument and high-performance components from BUCK Scientific
BUCK/Cecil Instruments
• Best optical design and highest quality components resulting in superior specifications:– Stray Light– Noise– Stability– Baseline Flatness– Accuracy (wavelength & photometric / ABS)– Reproducibility (wavelength & photometric / ABS)
BUCK/Cecil Series
• Series 2000
– Single Beam, Scanning, 4nm bandpass– Optional built-in printer & P-C Interface– Many software options– Most POPULAR Industrial system– Applications: Bio-Quest, Aqua-Quest, Food-
Quest series with Prep-Programmed Operations
BUCK/Cecil Series
• Series 3000– Single Beam, Scanning, 1.8nm bandpass– Optional built-in printer & P-C interface– Many software features (Kinetics, Bio-Assay)– Reflectance option: 3055 w/ Integrating Sphere
Specifically designed to meet ALL of the required Specifications for USP / BP
Pharmaceutical Assays
BUCK/Cecil Series
• Series 7000/7000+/9000– Double-Beam, Scanning, some Models with
variable 0.5 / 1.0 / 1.8-2.0 / 4.0nm bandpass
– Standard Printer & P-C interface, Data-Stream– Many software features (Kinetics, Bio-Assay)– Reflectance / Fluorescence / Turbidity options
Ideal for the stringent requirements of Academic and Pharmaceutical Research
Sample Handling Options
• Cuvette - standard cell holder– Quartz for UV range– Glass & Plastic for Vis range– 1-100mm Pathlengths
• Flow Cell - standard or micro flow (50L)– Uses Sipette Pump– Optional Automatic Cell Changer
Sample Handling Options
• Multi-Cell Changer– 4, 6, or 8 Positions– Manual, Automatic, Programmable
• Thermostatted Cell Holder– Water-jacketted or Electronic Peltier system
• Batch Sampler - 40 Positions Carousel• Specular & Integrating Sphere / Diffuse
Reflectance
Sample Handling Options
Multiple position CELL CHANGERS, MICRO-SIPPERS and Sequential Access BATCH SAMPLERSAllow the Chemist completeFLEXIBILITY for MAXIMUM Sample Throughput & Volumeon ANY of the BUCK/CecilUV-Vis Spectrophotometers
Software Features
A convenient INTERNAL Ink-Printer,and several P-C Software Programs provide many Data Processing and Manipulation options for Qualitative & Quantitative Work
Economical UV-Scan and sophisticated
GRAMS/UV & Data-Stream are available
Software Features
The INTERNAL Software built-in to the BUCK/Cecil UV-Vis Spectrophotometers is capable of performing General Analytical, Environmental, Biotechnology and Clinical Assay WITHOUT the need for a Computer:
• Christian-Warburg & Protein / DNA Measurements• Colorimetric Anion Calibrations stored in the instrument• Wavelength Subtraction, Spectral Overlay & Smoothing,
Derivatization and auto Peak-Pick on all systems• Multi-Component Analysis, with 1o / 2o and 3o Polynomial
Calibrations with up to 10 Standards with Statistics
APPLICATIONS
Primarily used for QUANTITATIVE Analysis of Known Compounds, UV-
Vis is one of the most popular techniques in the Pharmaceutical,
Foods and Paints Industries, as well as Water Laboratories
APPNOTE - Inorganic
SAMPLE: River Water for PHOSPHATE
PREPARATION: Digest w/ 10% HCl, add NH4MoO4 & Hydrazine reagent, boil till Blue color develops; measure at 810nm.
ANALYSIS: CE-2041 or VIS-100 w/ USEPA 2-pt Calibration
RESULTS: mg/L or PPM in Sample
APPNOTE - Inorganic
DATE Total-PO4 ortho-PO4
3-10-00 69 PPM 51 PPM
3-24-00 77 PPM 50 PPM
4-08-00 94 PPM 83 PPM
4-22-00 118 PPM 72 PPM
5-10-00 85 PPM 78 PPM
ANION assays by Colorimetry are the ACCEPTED Standard World-Wide & much better (and less
expensive) than Ion-Chromatography
APPNOTE - Cosmetic
SAMPLE: Color-Dyes for Hair Shampoos
PREPARATION: Make 1:5 and 1:200 dilution of Dyes to compare to Samples
ANALYSIS: CE-2041 or -3041 Scanning Spectrophotometer,
1cm. Glass Cuvette
RESULT: % Red / Green / Blue based on Color Standards
APPNOTE - Nutritional
SAMPLE: Cereals and Grains (preserved)
PREPARATION: Hydrolyze 2 grams w/ 5% HCl, extract w/ Hexane for 1:10 dilution
ANALYSIS: CE-2041 or CE-3041 Scanning
Spectrophotometer, scan range 200-450nm. For BHT, BHA and TBHQ Anti-Oxidants. Sample solutions run in 10mm. Quartz Cells
RESULT: Parts-Per-Million TOTAL Anti-Oxidants based on USFDA Standard Procedure
APPNOTE - Nutritional
Sample BHA BHT TBHQ [Wavelength 410nm. 360nm. 285nm.]
Frosted Flakes 35 PPM 27 PPM 11 PPM Raisin Bran 16 PPM 41 PPM < 2 PPM Rice Krispies 23 PPM 11 PPM 4.5 PPM
Scanning SPECTROPHOTOMETRY is an ideal method to evaluate YOUR OWN products, or material
from the Competition
Other SPECTRO Analyzers from BUCK
BUCK Scientific also offers a series of inexpensive, high-performing
and reliable Analytical Spectroscopy equipment for special
Applications, Educational and Dedicated Testing procedures
FLUORESCENCE SPECTROSCOPY (FLUOR)
FLUORESCENCE is a form of Molecular “EMISSION” Spectroscopy; where a Sample in a
special Cuvette is exposed to invisible, high-Energy UV radiation, and certain parts of the
Organic Molecule will “give off” or EMIT lower-Energy VISIBLE Light radiation.
Very specific & selective test for Drugs & Vitamins, Alkaloids & Toxins
Theory & Principle of OPERATION
BUCK Filter Fluorometer
The advantage of HIGH Sensitivity for most compatible materials makes Fluorescence a valuable analytical tool for Forensic & Drug applications
Intense BLUE Fluorescence of
QUININE in a Drug Formulation
BUCK Filter Fluorometer
The availability of a wide assortment of Excitation and Emission Filters makes this portable FL-380 instrument very flexibility for a very low cost… ideal for Academic Teaching & Industrial QA/QC work
Visible COLORIMETRY (COLOR)
This is for use in Academic Teaching classes and for dedicated COLORIMETRIC
Analyses for materials such as ANIONS (SO4, NO2, NO3, PO4, etc) and Visible Colors of compound such as Pigments,
Chlorophyll and Dyes
Covers the range from 320 - 950nm.
Theory & Principle of OPERATION
BUCK Colorimeter
The VIS-100 is a small, durable, self-calibrating unit that provides MEDIUM resolution @ 8nm. Bandpass, automatic 1-point Calibration & Data Outputs for a very low price