Determination of serum cholestanol by semi-micro high-performance liquid chromatography with electrochemical detection.

A simple and sensitive method that dose not require derivatization for determining cholestanol has been developed using HPLC with electrochemical detection (HPLC-ECD). The current peak height was linearly related to the amount of cholestanol injected, ranging from 1 to 200 muM (r = 0.999). The detection limit (S/N = 3) of cholestanol was 0.23 muM (1.2 pmol). Total cholestanol in control human and mouse serum was determined by the present method with a recovery rate of more than 90% and an RSD (n = 5) of less than 7.3%. Further, this method was successfully applied to monitor experimental hypercholestanolemia in mice fed a high-cholestanol diet, an animal model of cerebrotendinous xanthomatosis (CTX). In conclusion, we found this method to be both simple and useful for the determination of cholestanol in serum, helping in the diagnosis of CTX.