This basic spectrophotometer training session should enable you to know: Basics of how a Spec’ Works What the Individual Components Actually Do Commonly used Terminology

Deuterium / D2 Tungsten Halogen Xenon Lamp Covers Whole Range

Filter Prism Grating

Photocell Photodiode Photomultiplier

If SBW of instrument is 8nm Eg. Set to 700nm  Also often referred to as:- Bandwidth, Bandpass, Slit Width, Resolution.

Optimum SBW is 1/10 th of the Natural Bandwidth of the peak

The effect of SBW on the ability to resolve spectral peaks

SBW Optics Source Lamp Entrance Slit Grating Exit Slit Sample Detector

Simple Fixed Filter Instruments Fixed Grating Instruments Scanning Grating Instruments - Fixed SBW Scanning Grating Instruments - Variable SBW

Multiple Absorbance Fixed Absorbance Fixed Concentration using Factor Fixed Concentration using Standard(s) Scanning Absorbance at Fixed v Time - Kinetics

Spectrum Regions - Typical Wavelengths* Ultra Violet180 nmto 400 nm 1800 Å to 4000 Å.18 µm to.4 µ m 55,556 cm-1 to 25,000 cm-1 ( 5.5 * 10 6 Hzto2.5 * 10 6 Hz ) Visible400 nm to800 nm 4000 Å to 8000 Å.400 µ mto.8 µ m 25,000 cm-1 to 12,500 cm-1 ( 2.5 * 10 6 Hzto1.25 * 10 6 Hz ) Near Infra Red 800 nmto 2500 nm 8000 Å to Å.8 µ mto2.5 µ m 12,500 cm-1to 4000 cm-1 (1.25 * 10 6 Hzto4 * 10 5 Hz ) Infra Red2,500 nm to 15,000 nm 25,000 Å to 150,000 Å 2.5 µ mto15 µ m 4,000 cm-1to 667 cm-1 ( 4 * 10 5 Hzto6.67 * 10 4 Hz ) Far Infra Red15,000 nmto 200,000 nm 150,000 Å to 2,000,000 Å 15 µ m to200 µ m 667 cm-1 to 50 cm-1 ( 6.67 * 10 4 Hzto5 * 10 3 Hz ) *Reference:*Dyer, John R Applications of Absorption Spectroscopy of Organic Compounds (2nd printing). Englewood Cliffs, NJ Prentice-Hall: pp 1,22.

Conversions Nanometer ( nm, millimicron ) 1 nm=1 x 10-9 meters Angstrom ( Å ) 1 Å=1 x meters Micron( µ m, micrometer ) 1 µ m=1 x 10-6 meters Wave Number ( cm-1 )1 cm-1 = # of waves per cm (Inverse Centimeter) To convert wave number to/from nm: nm =1 / (WaveNumber)(10-7) WaveNumber = 1 / ( nm )(10-7)

1cm Cell / Cuvette Single Holder, Multi-Cell Holder Long Pathlength Cell - Up to 10cm Test Tubes Flow Cells - Sipper System (Can link to Auto-sampler) Solid Sample Holders eg. For Filters

Easy Sampling - High Throughput

Cell Holder Flowcell Pump Sample inlet spout

 Academia & Teaching  Biological & Biochemical Research  Bio’ & Biochem’ Monitoring & Quality Control  Chemical Research  Chemical Process Monitoring & QC  Pharmaceutical Research  Pharmaceutical Monitoring & QC  Environmental Laboratories  Hospital Pharmacy & Clinical Laboratories  Industrial Laboratories  etc. etc. etc.  Basically just about ANY laboratory!

After a pH meter and a Balance, a Visible or UV-Vis Spectrophotometer is the next most common piece of equipment in most laboratories! Anything that goes near to, into, or is in contact with your body MUST either have been tested by or have had it’s components tested by a Visible or UV-Vis Spectrophotometer. Foods Beverages Pharmaceuticals Cosmetics Chemicals Fabrics Dyes Etc. etc. etc.

 What Wavelength(s) / Wavelength Range?  Type of Work?  Simple Absorbance  Concentration (Factor, Single / Multi Standard)  Scanning  Kinetics  Number of Methods  Number of Samples  Sample Volume  Sample Presentation Needs (Instrument screen, Printer, Computer)