A new method for determination of serum cholestanol by high-performance liquid chromatography with ultraviolet detection

We developed a method for the determination of serum 5α-cholestan-3β-ol (cholestanol). The sterols were derivatized to the 4′-bromobenzenesulfonyl esters and heated in isopropanol. The cholesterol-4′-bromobenzenesulfonate was solvolyzed to cholesteryl isopropyl ether, but the derivatized cholestanol did not change and could be measured in a high-performance liquid chromatographic system equipped with a UV detector at 235 nm. On the other hand, the resulting cholesteryl isopropyl ether, having different absorbance and chromatographic mobility was not detected. This method was used for measuring cholestanol levels in patients with cerebrotendinous xanthomatosis (CTX), liver cirrhosis and serum from healthy control subjects. Reproducibility, linearity and recovery tests were done on 0.3 ml of serum samples containing >2 μg/ml cholestanol, using stigmastanol as an internal standard (I.S.). Determining cholestenol by this method can be used for diagnosis and follow-up of patients with CTX and various liver diseases.