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Beer-Lambert Law, more commonly known as Beer's Law, states that the optical absorbance of a
chromophore in a transparent solvent varies linearly with both the sample cell pathlength and the
chromophore concentration. Beer's Law is the simple solution to the more general description of
Maxwell's far-field equations describing the interaction of light with matter. In practice, Beer's Law
is accurate enough for a range of chromophores, solvents and concentrations, and is a widely
used relationship in quantitative spectroscopy.
Aλ = -log (I/I0)
Aλ = ελbc
In an absorbance experiment, light is attenuated not only by the chromophore, but also by
reflections from the interface between air and the sample, the sample and the cuvette, and
absorbance by the solvent. These factors can be quantified separately, but are often removed by
defining I0 as the light passing through a sample "blank" or "baseline" or reference sample (for
example, a cuvette filled with solvent but zero concentration of the chromophore is used as the
blank).
Many factors can affect the validity of Beer's Law. It is usual to check for the linearity of Beer's
Law for a chromophore by measuring the absorbance of a series of standards. This "calibration"
can also remove errors in the experiment, the equipment, and the batch of reagents (such as
cuvettes of unknown pathlength).