1. Buck Scientific, Inc. Seminar in ANALYTICAL INSTRUMENTATION for Spectroscopy & Chromatography: UV-Vis Spectrophotometry

2. 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

3. 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

4. Basic SPECTROMETER Design ALL Analytical Spectrometers have several BASIC design components in common: Light Source Sample Chamber Optical System Detector Output

5. 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

6. UV/Vis Important Features Scan Speed Resolution Software Features Ease of Operation Data Storage Customized Calculations Support & Service

7. 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

8. 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)

9. 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

10. 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

11. 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

12. 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 (50 L) –Uses Sipette Pump –Optional Automatic Cell Changer

13. 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

14. Sample Handling Options Multiple position CELL CHANGERS, MICRO- SIPPERS and Sequential Access BATCH SAMPLERS Allow the Chemist complete FLEXIBILITY for MAXIMUM Sample Throughput & Volume on ANY of the BUCK/Cecil UV-Vis Spectrophotometers

15. 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

16. 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 1 o / 2 o and 3 o Polynomial Calibrations with up to 10 Standards with Statistics

17. 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

18. APPNOTE - Inorganic SAMPLE: River Water for PHOSPHATE PREPARATION: Digest w/ 10% HCl, add NH 4 MoO 4 & 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

19. APPNOTE - Inorganic DATETotal-PO4ortho-PO4 3-10-0069 PPM51 PPM 3-24-0077 PPM50 PPM 4-08-0094 PPM83 PPM 4-22-00118 PPM72 PPM 5-10-0085 PPM78 PPM ANION assays by Colorimetry are the ACCEPTED Standard World-Wide & much better (and less expensive) than Ion-Chromatography

20. 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

21. 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

22. APPNOTE - Nutritional SampleBHABHTTBHQ [Wavelength410nm.360nm.285nm.] Frosted Flakes35 PPM27 PPM11 PPM Raisin Bran16 PPM41 PPM< 2 PPM Rice Krispies23 PPM11 PPM4.5 PPM Scanning SPECTROPHOTOMETRY is an ideal method to evaluate YOUR OWN products, or material from the Competition

23. 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

24. 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

25. 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

26. 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

27. 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

28. 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