Ultraviolet. Spectrophotometry Method for the Determination of Cholesterol

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Abstract
A spectrophotometric method is described for the determination of cholesterol, either in its free form or as its digitonide. The technique is based upon the absorption of ultraviolet light in the 204 to 206 m[mu] region by the ethylene linkage in the B ring of cholesterol. The method is simple, 3 to 4 times more sensitive than the Schoenheimer-Sperry method, and considerably more reproducible. It is not markedly susceptible to the influence of time, humidity, concentration of reagents, temperature, or other variables to which the Schoenheimer-Sperry method is liable. The method is suitable for determining the free and total cholesterol in serum and tissues except in the presence of high concentrations of digitonin precipitable sterol vitamins and hormones. The cholesterol does not react with any reagents except digitonin and can be reclaimed.

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