- P-ISSN 2233-4203
- E-ISSN 2093-8950
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- P-ISSN2233-4203
- E-ISSN2093-8950
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Comparative GC-MS Based In vitro Assays of 5α-Reductase Activity Using Rat Liver S9 Fraction
Mass Spectrometry Letters / Mass Spectrometry Letters, (P)2233-4203; (E)2093-8950
2012, v.3 no.1, pp.21-24
https://doi.org/10.5478/MSL.2012.3.1.021
(Korea Institute of Science and Technology)
(Korea Institute of Science and Technolog)
(Korea Institute of Science and Technology)
(Yonsei University)
(Korea Institute of Science and Technology)
(Korea Institute of Science and Technolog)
(Korea Institute of Science and Technolog)
(Korea Institute of Science and Technology)
(Yonsei University)
(Korea Institute of Science and Technology)
(Korea Institute of Science and Technolog)
Su,
H.
L.
, Lee,
D., Lee,
J., Lee,
W., Chung,
B. C., &
Choi,
M.
H.
(2012). Comparative GC-MS Based In vitro Assays of 5α-Reductase Activity Using Rat Liver S9 Fraction. Mass Spectrometry Letters, 3(1), 21-24, https://doi.org/10.5478/MSL.2012.3.1.021
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Abstract
5α-Dihydrotestosterone (DHT) is the primary active metabolite of testosterone, catalyzed by 5α-reductase (5αR) in the skin, prostate, and liver. In this study, the 5αR 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 5αR inhibitor, finasteride, were added to the S9 fractions and incubated at 37oC 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 μM, whereas the isotope-dilution GC-MS (GC-IDMS) system was able to evaluate the 5αR 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 5αR activity in large-scale clinical studies.
- keywords
- 5α-Reductase, Testosterone, Dihydrotestosterone, Isotope-dilution, GC-MS, Liver microsome
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- Submission Date
- 2012-03-01
- Revised Date
- 2012-03-12
- Accepted Date
- 2012-03-12
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