Simultaneous quantitative determination of urinary cholesterol, desmosterol and lanosterol in pravastatin treated rats by gas chromatography/mass spectrometry

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

  • Kumar, Bhowmik Salil (Bioanalysis and Biotransformation Research Center, Korea Institute of Science and Technology) ;
  • Chung, Bong Chul (Bioanalysis and Biotransformation Research Center, Korea Institute of Science and Technology) ;
  • Lee, Young-Joo (College of Pharmacy, Kyung Hee University) ;
  • Yi, Hong Jae (College of Pharmacy, Kyung Hee University) ;
  • Jung, Byung Hwa (Bioanalysis and Biotransformation Research Center, Korea Institute of Science and Technology)
  • 사릴 꾸마르 (한국과학기술연구원 생체대사연구센터) ;
  • 정봉철 (한국과학기술연구원 생체대사연구센터) ;
  • 이영주 (경희대학교 약학대학) ;
  • 이홍재 (경희대학교 약학대학) ;
  • 정병화 (한국과학기술연구원 생체대사연구센터)
  • Received : 2009.07.09
  • Accepted : 2009.08.31
  • Published : 2009.10.25

Abstract

A simultaneous determination method for cholesterol, lanosterol and desmosterol was developed using gas chromatography/mass spectrometry. Urine was enzymatically hydrolyzed with ${\beta}$-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 ($r^2=0.998{\sim}0.999$) in calibration curve and a satisfactory recovery (80.0%~113%) were achieved. Accuracy and precision values within ${\pm}15%$ in the concentration range of 5 to 200 ng/mL were also observed for all compounds. The developed method was applied to pravastatin-treated (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.

Acknowledgement

Supported by : Korea Food & Drug Administration