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Beer and Lambert's law states that 'when a ray of monochromatic light passes through a solution in a transparent vessel, intensity of transmitted light depends on concentration of absorbing solution and path length of absorbing medium.'

A=ebc

Where A is absorbance (no units, since A = log10P0 / P )

e is the molar absorbtivity with units of L mol-1 cm-1

b is the path length of the sample - that is, the path length of the cuvette in which the sample is contained. We will express this measurement in centimetres.

c is the concentration of the compound in solution, expressed in mol L-1

when path length is kept constant ansorbance is proportional to concentration of substance. Path length for a colorimeter is constant.

Different substances absorb light of different frequencies maximally. Light of appropriate frequency is passed through solution using different filters in colorimeter and % transmission is measured. e.g. proteins absorb light of 650nm.

Error is eliminated by calliberatin blank(solution without test substance) as 100% transmission ie zero absorbance, and black stop as 0% transmission. Thus we get absorbance of only required substance.

% transmission is converted to absorbance using following expression

A = 2 - log10 %T

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