Gas Chromatography/Mass Spectrometry를 이용한 Pravastatin 투여 쥐의 뇨 중 Cholesterol, Desmosterol, Lanosterol의 동시분석법

Abstract

A simultaneous determination method for cholesterol, lanosterol and desmosterol was developed using gas chromatography/mass spectrometry. Urine was enzymatically hydrolyzed with β-glucuronidase/arylsulfatase. Samples were prepared using extractions with a mixture of ethyl acetate-hexane (2:3, v/v), followed by derivatization with a mixture of MSTFA/TMSI/TMCS (100:2:5 v/v/v). All analyses were performed using GC/ MS in selective ion monitoring mode. Good linearities (r2=0.998~0.999) in calibration curve and a satisfactory recovery (80.0%~113%) were achieved. Accuracy and precision values within ±15% in the concentration range of 5 to 200 ng/mL were also observed for all compounds. The developed method was applied to pravastatintreated (70 and 250 mg/kg/day for 7 days, oral) hyperlipidemia rats. Those sterols were significantly lower in drug-treated rats compared to the controls, which justifies the drug efficacy. Therefore, these results indicate that the developed method was successfully applied to examine statin drug efficacy with urine sample.