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