Development and validation of a sensitive LC/MS/MS method for the simultaneous determination of naloxone and its metabolites in mouse plasma

详细信息    查看全文

• 作者：Hongliang Jiang^a ; ^b ; ^{jianghlf@gmail.com} ; YuRong Wang^b ; Manjunath S. Shet^c ; Yang Zhang^b ; Duane Zenke^b ; Douglas M. Fast^b
• 关键词：Naloxone ; 6¦Â-Naloxol ; Naloxone-3¦Â-d-glucuronide ; LC&ndash ; MS/MS ; Quantification
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2011
• 出版时间：1 September, 2011
• 年：2011
• 卷：879
• 期：25
• 页码：2663-2668
• 全文大小：1K

文摘

A rapid, specific, and reliable LC-MS/MS based bioanalytical method was developed and validated for the simultaneous determination of naloxone (NLX) and its two metabolites, 6¦Â-naloxol (NLL) and naloxone-3¦Â-d-glucuronide (NLG) in mouse plasma. The optimal chromatographic behavior of these analytes was achieved on an Aquasil C18 column (50 mm ¡Á 2.1 mm, 5 ¦Ìm) using reversed phase chromatography. The total LC analysis time per injection was 2.5 min with a flow rate of 1.0 mL/min with gradient elution. Sample preparation via protein precipitation with acetonitrile in a 96-well format was applied for analyses of these analytes. The analytes were monitored by electrospray ionization in positive ion multiple reaction monitoring (MRM) mode. Modification of collision energy besides chromatographic separation was applied to further eliminate interference peaks for NLL and NLG. The method validation was conducted over the curve range of 0.200/0.400/0.500 to 100/200/250 ng/mL for NLX/NLL/NLG, respectively, using 0.0250 mL of plasma sample. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels showed ?.5 % relative standard deviation (RSD) and ?.3 to ?.5 % relative error (RE). The method was successfully applied to determine the concentrations of NLX, NLL, and NLG in incurred mouse plasma samples.