Ethyl acetate extraction procedure and isocratic high-performance liquid chromatographic assay for testosterone metabolites in cell microsomes
- PMID: 9551806
- DOI: 10.1016/s0378-4347(97)00565-3
Ethyl acetate extraction procedure and isocratic high-performance liquid chromatographic assay for testosterone metabolites in cell microsomes
Abstract
An isocratic reversed-phase high-performance liquid chromatography (HPLC) method was developed and validated for separation of testosterone and its main metabolites over the nominal range 20 to 40 microg/ml and 280 to 4600 ng/ml, respectively. Mobile phase composition (phosphate buffer-methanol-acetonitrile, 50:38.5:11.5) was optimised by studying the influence of numerous chromatographic parameters. The most critical one was the ratio CH3CN/CH3OH. Good recoveries (around 90% for all compounds) and an improved specificity were assessed by a double ethyl acetate extraction of biological samples. According to the performance criteria tested, the method could be applied to enzymatic inhibition and induction in vitro studies.
Similar articles
-
Rapid high-performance liquid chromatographic method for the separation of hydroxylated testosterone metabolites.J Chromatogr B Biomed Sci Appl. 2001 Sep 5;760(2):281-8. doi: 10.1016/s0378-4347(01)00286-9. J Chromatogr B Biomed Sci Appl. 2001. PMID: 11530987
-
[Determination of cytochrome P450 3A4 activity with testosterone probe using high performance liquid chromatography].Se Pu. 2008 Jan;26(1):80-3. Se Pu. 2008. PMID: 18438030 Chinese.
-
Simultaneous determination of testosterone metabolites in liver microsomes using column-switching semi-microcolumn high-performance liquid chromatography.Anal Biochem. 2001 Aug 15;295(2):248-56. doi: 10.1006/abio.2001.5223. Anal Biochem. 2001. PMID: 11488629
-
Isocratic high-performance liquid chromatographic method for the separation of testosterone metabolites.J Chromatogr B Biomed Appl. 1995 Oct 20;672(2):207-15. doi: 10.1016/0378-4347(95)00216-6. J Chromatogr B Biomed Appl. 1995. PMID: 8581126
-
Hydroxylation of 5 alpha-androstane-3 beta,17 beta-diol by rat prostate microsomes: effects of antibodies and chemical inhibitors of cytochrome P450 enzymes.Arch Biochem Biophys. 1992 Aug 1;296(2):355-65. doi: 10.1016/0003-9861(92)90585-k. Arch Biochem Biophys. 1992. PMID: 1632629
Cited by
-
Antimicrobial, Quorum Sensing Inhibition, and Anti-Cancer Activities of Silver Nanoparticles Synthesized from Kenyan Bacterial Endophytes of Teclea nobilis.Int J Mol Sci. 2025 Apr 2;26(7):3306. doi: 10.3390/ijms26073306. Int J Mol Sci. 2025. PMID: 40244167 Free PMC article.
-
Oral Contraceptives Induce Time- and Intestinal Segment-Dependent Shifts in the Gut Microbiota.Nutrients. 2025 Aug 9;17(16):2591. doi: 10.3390/nu17162591. Nutrients. 2025. PMID: 40871620 Free PMC article.
-
Moduratory effect of Thai traditional medicine (Yahom Tultavai) on hepatic cytochrome P450 enzymes and pentobarbital-induced sleeping in mice.Afr J Tradit Complement Altern Med. 2013 May 16;10(4):128-36. doi: 10.4314/ajtcam.v10i4.21. eCollection 2013. Afr J Tradit Complement Altern Med. 2013. PMID: 24146513 Free PMC article.