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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
2
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pubmed:dateCreated |
2010-5-25
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pubmed:abstractText |
A novel method was established for simultaneous quantitation of testosterone (T) and dihydrotestosterone (DHT) in murine tissue and serum samples. Endogenous T and DHT, together with the internal standards 17alpha-methyl-T and 17alpha-methyl-DHT, were extracted from tissues and then derivatized by reaction with 2-hydrazino-4-(trifluoromethyl)-pyrimidine (HTP). Analysis by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) resulted in product ion spectra of HTP derivatives of both T and DHT that showed analyte-specific fragmentations; the latter fragmentations were characterized by the use of high-resolution Orbitrap MS/MS. These specific fragmentations enabled quantitation of T and DHT in the multiple-reaction monitoring (MRM) mode. The method was validated with charcoal-stripped serum as the matrix. The lower limit of quantitation (LLOQ) was 0.10ng/ml for T and 0.50ng/ml for DHT. The method was then used for determination of serum and tissue levels of T and DHT in transgenic mice carrying a hypomorphic NADPH-cytochrome P450 reductase gene (Cpr-low mice). Remarkably, ovarian T levels in Cpr-low mice were found to be 25-fold higher than those in wild-type mice, a finding that at least partly explains the female infertility seen in the Cpr-low mice. In conclusion, our method provides excellent sensitivity and selectivity for determination of endogenous levels of T and DHT in mouse tissues.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/20361922-10214723,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/20361922-9709942
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
1096-0309
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pubmed:author |
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pubmed:copyrightInfo |
2010 Elsevier Inc. All rights reserved.
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pubmed:issnType |
Electronic
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pubmed:day |
15
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pubmed:volume |
402
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
121-8
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pubmed:dateRevised |
2011-8-1
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pubmed:meshHeading |
pubmed-meshheading:20361922-Animals,
pubmed-meshheading:20361922-Chromatography, Liquid,
pubmed-meshheading:20361922-Dihydrotestosterone,
pubmed-meshheading:20361922-Female,
pubmed-meshheading:20361922-Male,
pubmed-meshheading:20361922-Mice,
pubmed-meshheading:20361922-Mice, Inbred C57BL,
pubmed-meshheading:20361922-Mice, Transgenic,
pubmed-meshheading:20361922-NADPH-Ferrihemoprotein Reductase,
pubmed-meshheading:20361922-Spectrometry, Mass, Electrospray Ionization,
pubmed-meshheading:20361922-Testosterone,
pubmed-meshheading:20361922-Tissue Distribution
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pubmed:year |
2010
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pubmed:articleTitle |
Analysis of testosterone and dihydrotestosterone in mouse tissues by liquid chromatography-electrospray ionization-tandem mass spectrometry.
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pubmed:affiliation |
Wadsworth Center, New York State Department of Health, State University of New York at Albany, 12201, USA.
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pubmed:publicationType |
Journal Article,
Research Support, N.I.H., Extramural,
Validation Studies
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