Box 31 Working In Biochemistry

Absorption of Light by Molecules: The Lambert-Beer Law

A wide range of biomolecules absorb light at characteristic wavelengths, just as tryptophan absorbs light at 280 nm (see Fig. 3-6). Measurement of light absorption by a spectrophotometer is used to detect and identify molecules and to measure their concentration in solution. The fraction of the incident light absorbed by a solution at a given wavelength is related to the thickness of the absorbing layer (path length) and the concentration of the absorbing species (Fig. 1). These two relationships are combined into the Lambert-Beer law,

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