Simultaneous determination of β-sitosterol, campesterol, stigmasterol, and brassicasterol in serum by high-performance liquid chromatography with electrochemical detection

摘要

A simple method for the determination of phytosterols such as β-sitosterol, campesterol, stigmasterol, and brassicasterol has been developed using high-performance liquid chromatography with electrochemical detection (HPLC-ECD). Under optimized conditions, the current peak height was linearly related to the amount of phytosterol injected, ranging from 10 to 200 μmol L?1 (r = 0.999). The relative standard deviation (RSD) was less than 3.1% (100 μmol L?1, n = 6). The detection limit (S/N = 3) of phytosterol was less than 3.4 μmol L?1 (17 pmol). Phytosterols spiked into the control human and rat sera were determined by the present method with recovery rates of more than 80% and RSDs (n = 3) of less than 4.2%. This method has been successfully applied to the monitoring of experimental phytosterolemia in rats induced by a high-phytosterol diet. Taken together, we found this method, which does not require a derivatization step, is both simple and useful for the simultaneous determination of serum phytosterols, providing an alternative tool for the diagnosis of phytosterolemia...