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The SI unit of molar absorption coefficient is the square metre per mole (m 2 /mol), but in practice, quantities are usually expressed in terms of M −1 ⋅cm −1 or L⋅mol −1 ⋅cm −1 (the latter two units are both equal to 0.1 m 2 /mol).
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These two different types of hemoglobin exhibit different absorption spectra that are normally represented in terms of molar extinction coefficients, as shown in Figure 1. The molar extinction coefficient of Hb has its highest absorption peak at 420 nm and a second peak at 580 nm. Its spectrum then gradually decreases as light wavelength increases.
Extinction coefficient refers to several different measures of the absorption of light in a medium: Attenuation coefficient , sometimes called "extinction coefficient" in meteorology or climatology Mass extinction coefficient , how strongly a substance absorbs light at a given wavelength, per mass density
First step is to plot the absorbance(A) values of standard solution against molar concentrations (c) of the known solution. Then the best straight line is plotted, passing through the origin. The experimental points are plotted as per Beer’s law: A= E*c*l where E= molar extinction coefficient and l= optical path length usually 1 cm.
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Variable pathlength absorption spectroscopy uses a determined slope to calculate concentration. As stated above this is a product of the molar absorptivity and the concentration. Since the actual absorbance value is taken at many data points at equal intervals, background subtraction is generally unnecessary.
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