Simultaneous determination of oxymorphone and its active metabolite 6-OH-oxymorphone in human plasma by high performance liquid chromatography-tandem mass spectrometry

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• 作者：Wuyi Zha ; Linyee Shum
• 关键词：Oxymorphone ; 6-OH-oxymorphone ; LC&ndash ; MS/MS ; SPE ; Human plasma
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2012
• 出版时间：1 August, 2012
• 年：2012
• 卷：902
• 期：Complete
• 页码：116-121
• 全文大小：352 K

文摘

A selective high performance liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the simultaneous determination of oxymorphone and its active metabolite 6-OH-oxymorphone in human plasma was developed and validated using oxymorphone-d₃ as the internal standard. Chromatographic conditions were optimized to separate oxymorphone from the other metabolite, oxymorphone-3-glucuronide, which may convert to oxymorphone in MS ion source, resulting in inaccurate quantitation of oxymorphone. Solid phase extraction (SPE) was used to extract oxymorphone and 6-OH-oxymorphone from plasma. SPE offered the advantage of being able to remove the unwanted metabolite, oxymorphone-3-glucuronide, through the wash step during the extraction. The developed method was precise and reproducible as shown by good linearity of calibration curves (correlation coefficients ¡Ý0.9968 for oxymorphone and ¡Ý0.9967 for 6-OH-oxymorphone) with high intraday assay and interday assay precision (CV % ¡Ü11.0 % for oxymorphone and ¡Ü12.6 % for 6-OH-oxymorphone) over a range of 35/25 - 5000/5000 pg/mL for oxymorphone/6-OH-oxymorphone. The method has been successfully applied to analyze oxymorphone and 6-OH-oxymorphone in plasma from 19 healthy volunteers in a bioequivalence study. A total of 1026 samples were analyzed. Good linearity (average correlation coefficient 0.9988 for oxymorphone and 0.9966 for 6-OH-oxymorphone) was achieved with calibration curves and high precision (CV % ¡Ü5.9 % for oxymorphone and ¡Ü10.9 % for 6-OH-oxymorphone) was obtained with QCs.