Linked Life Data

20361922

Source:http://linkedlifedata.com/resource/pubmed/id/20361922

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pubmed-article:20361922pubmed:abstractTextA 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.lld:pubmed
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pubmed-article:20361922pubmed:authorpubmed-author:LinN CNClld:pubmed
pubmed-article:20361922pubmed:authorpubmed-author:SpinkDavid...lld:pubmed
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pubmed-article:20361922pubmed:authorpubmed-author:XieFangFlld:pubmed
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pubmed-article:20361922pubmed:copyrightInfo2010 Elsevier Inc. All rights reserved.lld:pubmed
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pubmed-article:20361922pubmed:dateRevised2011-8-1lld:pubmed
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pubmed-article:20361922pubmed:articleTitleAnalysis of testosterone and dihydrotestosterone in mouse tissues by liquid chromatography-electrospray ionization-tandem mass spectrometry.lld:pubmed
pubmed-article:20361922pubmed:affiliationWadsworth Center, New York State Department of Health, State University of New York at Albany, 12201, USA.lld:pubmed
pubmed-article:20361922pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:20361922pubmed:publicationTypeResearch Support, N.I.H., Extramurallld:pubmed
pubmed-article:20361922pubmed:publicationTypeValidation Studieslld:pubmed