Determination of dihydrocholesterol in serum

• Published in: Analytical biochemistry Vol. 10; no. 3; pp. 435 - 443
• Main Authors: Chattopadhyay, D.P., Mosbach, E.H.
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01.01.1965
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/0003-2697(65)90312-X

1. (1) An isotope derivative procedure was developed for the determination of dihydrocholesterol (5α-cholestan-3β-ol) in 0.2 to 1.0 ml of serum. The method is based on the finding that the serum sterols, consisting predominantly of cholesterol and dihydrocholesterol, react quantitatively with acetic-1-C 14 anhydride and that the acetates are separated completely and rapidly by thin-layer chromatography on AgNO 3-silicic acid plates. The ratio of the radioactivities of the acetates will then correspond to the molar ratio of cholesterol to dihydrocholesterol in the sample. 2. (2) The feasibility of the method was tested by analyses of known cholesterol and dihydrocholesterol mixtures and by recovery experiments. The reproducibility of the procedure was checked by replicate analyses of serum from different species of animals. 3. (3) Serum dihydrocholesterol concentrations were determined in 7 species of laboratory animals maintained on a stock diet. The concentration of dihydrocholesterol ranged from 0.093 mg per 100 ml in the rat to 1.43 mg per 100 ml in the monkey. Chicken, guinea pig, dog, and mouse exhibited intermediate values.