Buck Scientific, Inc.

Seminar in ANALYTICAL
INSTRUMENTATION for
Spectroscopy & Chromatography:
UV-Vis Spectrophotometry
Ultra-Violet/Visible SPECTROSCOPY (UV-Vis)

Theory & Principle of OPERATION

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
 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 (50L)
– 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 SAMPLERS
Allow the Chemist complete
FLEXIBILITY for
MAXIMUM Sample
Throughput & Volume
on ANY of the BUCK/Cecil
UV-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)

Theory & Principle of OPERATION

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
 BUCK Filter Fluorometer

The advantage of
HIGH Sensitivity Intense BLUE
Fluorescence of
for most compatible QUININE in a Drug
materials makes Formulation
Fluorescence a
valuable analytical
tool for Forensic &
Drug applications
 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)

Theory & Principle of OPERATION

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.

 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