Simultaneous determination of norethisterone and six metabolites in human plasma by capillary gas chromatography with mass-selective detection |
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Authors: | F. Pommier A. Sioufi J. Godbillon |
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Affiliation: | 1. Food and Bioengineering Department, Henan University of Science and Technology, Luoyang, Henan 471003, China;2. College of Life Science, Yantai University, Yantai, Shandong 264025, China;1. Laboratory of Research in Neuroscience (LAREN), Pôle Technologie Santé (PTS), Faculty of Medicine, Saint Joseph University of Beirut, Beirut, Lebanon;2. Department of Neurosurgery – Hôtel-Dieu de France Hospital, Beirut, Lebanon |
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Abstract: | A method for the simultaneous determination of norethisterone (NET) and six metabolites in human plasma by capillary gas chromatography-mass-selective detection (GC-MS) is described. The compounds are determined in plasma after enzymatic hydrolysis. After addition of norgestrel as the internal standard, the compounds are extracted from plasma at pH 5 using an Extrelut column and elution with dichloromethane. After evaporation, the compounds are converted into bistrimethylsilyl derivatives which are determined by gas chromatography using a mass-selective detector at m/z 429 for the two dihydro-NET (5β-NET and 5α-NET), m/z 431 for the four tetrahydro-NET (3α,5α-NET, 3α,5β-NET, 3β,5β-NET and 3β,5α-NET), m/z 442 for NET and m/z 456 for the internal standard. The reproducibility and accuracy of the method were found suitable over the range of concentrations between 0.50 and 8 ng/ml for NET, and metabolites except for 5α-dihydro-NET (between 1 and 8 ng/ml). The method was applied to clinical samples. |
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Keywords: | Norethisterone Dihydronorethisterone Tetrahydronorethisterone |
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