A method for the determination of cholesterol in animal tissues by HPL
C is described. The extraction of cholesterol(esters) from the sample
is achieved by a mixture of hexane and isopropanol (3:2), which has be
en proven as the best procedure with the help of radioactive tracers.
In an aliquot of the extract a methanolysis of the cholesterolesters i
s made; interfering substances are removed on a silicagel-column, whic
h was standardized in its activity by addition of 10% water. Cholester
ol is eluted from the silica-column by dichloromethan. The chromatogra
phic analysis is performed isocratically on a RP18-column by a mixture
of acetonitrile and isopropanol (80:20) measuring at a wavelength of
210 nm. During the entire analysis stigmasterol as internal standard i
s used.