Mass Spectrometry Letters

Korean Society for Mass Spectrometry (한국질량분석학회)
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Comparative GC-MS Based In vitro Assays of 5α-Reductase Activity Using Rat Liver S9 Fraction

  • Lee, Su-Hyeon (Future Convergence Research Division, Korea Institute of Science and Technology) ;
  • Lee, Dong-Hyoung (Future Convergence Research Division, Korea Institute of Science and Technology) ;
  • Lee, Jeong-Ae (Future Convergence Research Division, Korea Institute of Science and Technology) ;
  • Lee, Won-Yong (Department of Chemistry, Yonsei University) ;
  • Chung, Bong-Chul (Future Convergence Research Division, Korea Institute of Science and Technology) ;
  • Choi, Man-Ho (Future Convergence Research Division, Korea Institute of Science and Technology)
  • Received : 2012.03.01
  • Accepted : 2012.03.12
  • Published : 2012.03.15
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Abstract

$5{\alpha}$-Dihydrotestosterone (DHT) is the primary active metabolite of testosterone, catalyzed by $5{\alpha}$-reductase ($5{\alpha}R$) in the skin, prostate, and liver. In this study, the $5{\alpha}R$ activity in rat liver S9 fraction in the presence of a NADPH-generating system was evaluated and compared by gas chromatography-mass spectrometry (GC-MS)-based in vitro assays. Testosterone and a $5{\alpha}R$ inhibitor, finasteride, were added to the S9 fractions and incubated at $37^{\circ}C$ for 1 h. Both testosterone and DHT were quantitatively measured and compared with two different GC-MS-based steroid profiling techniques. DHT was not detected by conventional GC-MS analysis in the absence of finasteride when the concentration of testosterone in the S9 fraction was less than $0.2{\mu}M$, whereas the isotope-dilution GC-MS (GC-IDMS) system was able to evaluate the $5{\alpha}R$ activity. Because the S9 fraction contains more reactive enzymes and is easier to collect from tissues compared with a microsomal solution, the combination of the S9 fraction and GC-IDMS technique may be a promising assay for evaluating the $5{\alpha}R$ activity in large-scale clinical studies.

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