CHOLESTEROL - ITS DETERMINATION IN MUSCLE AND ADIPOSE-TISSUE AND IN OFFAL USING HPLC

A method of determining the cholesterol content of animal tissues by m eans of HPLC is described. The cholesterol (esters) are extracted from the sample material with a mixture of hexane and isopropanol (3:2) wh ich has been found particularly suitable in experiments with labelled compounds. Methanolysis of the cholesterol esters is carried out in an aliquot of the extract. Disruptive substances are then separated off from the cholesterol on a silica gel column with its activity adjusted by the addition of 10% water. Dichloromethane is used to elute the ch olesterol from the column. Chromatography is done isocratically on an RP18 column with a mixture of acetonitrile and isopropanol (80:20) at a measurement wavelength of 210 nm. Stigmasterol is used as internal s tandard during the whole analytical procedure. Quantitative extraction with hexane/isopropanol, the correction of possible losses by the int ernal standard and simple chromatographic conditions give a good guara ntee of reliable analytical results with this method.