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Gang Street

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REFERENCES

1) http://chemed.chem.purdue.edu/genchem/lab/equipment/pipet/error.html

2) http://www.indiana.edu/~nimsmsf/P215/p215notes/LabManual/Lab3.pdf

3) http://www.biochem.duke.edu/wysiwyg/downloads/GrimsleyPace.pdf

4) http://www.clinchem.org/content/21/6/776.full.pdf

INTRODUCTION
Basic Laws of Light Absorption.
For a uniform absorbing medium (solution: solvent and solute molecules that absorb light) the proportion of light radiation passing through it is called the transmittance, T, and the proportion of light absorbed by molecules in the medium is absorbance, Abs. Transmittance is defined as:

T = I/Io where: Io= intensity of the incident radiation entering the medium. I = intensity of the transmitted radiation leaving the medium.

T is usually expressed as percent transmittance, %T:

%T = I/Io x 100

The relationship between percent transmittance (%T) and absorbance (A) is given by the following equation:

A = 2-log (%T)

On most spectrophotometers two scales are present, %T and Abs. Absorbance has no units (Why?) and varies from 0 to 2 (linear region for most substances is from 0.05 to 0.7). The Beer-Lambert Law states that Abs is proportional to the concentration (c) of the absorbing molecules, the length of light-path through the medium and the molar extinction coefficient:
A = εcl where: ε = molar extinction coefficient for the absorbing material at wavelength in units of 1/(mol x cm) c = concentration of the absorbing solution (molar) l = light path in the absorbing material (l=1 cm for our purposes)

The Beer-Lambert Law may not be applicable to all solutions since solutions can ionize/polymerize at higher concentrations, or precipitate to give a turbid suspension that may increase or decrease the apparent absorbance. Further, the Beer-Lambert Law is most accurate between Abs

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