Selective and sensitive liquid chromatography–tandem mass spectrometry method for the determination of levonorgestrel in human plasma

A selective, sensitive and rapid liquid chromatography–tandem mass spectrometry method for the determination of levonorgestrel in plasma was developed. An Applied Biosystems API 3000 triple quadrupole mass spectrometer set to multiple reaction monitoring (MRM) mode, using atmospheric pressure photos...

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Bibliographic Details
Published in:	Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 813; no. 1; pp. 331 - 336
Main Authors:	Theron, H.B., Coetzee, C., Sutherland, F.C.W., Wiesner, J.L., Swart, K.J.
Format:	Journal Article
Language:	English
Published:	Amsterdam Elsevier B.V 25-12-2004 Elsevier Science
Subjects:	17-α-Methyltestosterone Analysis Analytical, structural and metabolic biochemistry Atmospheric pressure photospray ionisation (APPI) Biological and medical sciences Calibration Chromatography, Liquid - methods Contraceptive Agents, Female - blood Fundamental and applied biological sciences. Psychology General pharmacology Humans Levonorgestrel Levonorgestrel - blood Liquid chromatography Liquid–liquid extraction Mass spectrometry Mass Spectrometry - methods Medical sciences Pharmacology. Drug treatments Plasma Reference Standards Sensitivity and Specificity Plasma Atmospheric pressure photospray ionisation (APPI) Levonorgestrel 17-α-Methyltestosterone Liquid–liquid extraction Liquid chromatography Mass spectrometry Human Biological fluid Mass spectrometry MS/MS Contraceptive Progestagen Quantitative analysis Blood plasma
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Summary:	A selective, sensitive and rapid liquid chromatography–tandem mass spectrometry method for the determination of levonorgestrel in plasma was developed. An Applied Biosystems API 3000 triple quadrupole mass spectrometer set to multiple reaction monitoring (MRM) mode, using atmospheric pressure photospray ionisation (APPI) in the positive mode. Using 17-α-methyltestosterone as internal standard (IS), liquid–liquid extraction was followed by reversed phase liquid chromatography using a phenyl–hexyl column and tandem mass spectrometric detection. The mean recovery for levonorgestrel and 17-α-methyltestosterone was 99.5 and 62.9%, respectively. The method was validated from 0.265 to 130 ng levonorgestrel/ml plasma with the lower limit of quantification (LLOQ) set at 0.265 ng/ml. This assay method makes use of the increased sensitivity and selectivity of tandem mass spectrometric (MS/MS) detection, allowing for a rapid (extraction and chromatography) and selective method for the determination of levonorgestrel in human plasma. The assay method was used in a pharmacokinetic study to quantify levonorgestrel in human plasma samples generated after administrating a single oral dose of 1.5 mg levonorgestrel to healthy female volunteers for up to five half lives. The total chromatographic runtime of this method was 5.0 min per sample, allowing for analysis of a large number of samples per batch.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1570-0232 1873-376X
DOI:	10.1016/j.jchromb.2004.10.039