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Simultaneous analysis of gemfibrozil, morphine, and its two active metabolites in different mouse brain structures using solid‐phase extraction with ultra‐high performance liquid chromatography and tandem mass spectrometry with a deuterated internal standard
Author(s) -
Yang Zizhao,
Wang Lu,
Xu Mingcheng,
Gu Jingkai,
Yu Lushan,
Zeng Su
Publication year - 2016
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201600088
Subject(s) - chemistry , chromatography , solid phase extraction , liquid chromatography–mass spectrometry , morphine , formic acid , extraction (chemistry) , mass spectrometry , tandem mass spectrometry , high performance liquid chromatography , detection limit , pharmacology , medicine
A rapid and sensitive bioassay was established and validated to simultaneously determine gemfibrozil, morphine, morphine‐3β‐glucuronide, and morphine‐6β‐glucuronide in mouse cerebrum, epencephalon, and hippocampus based on ultra‐high performance liquid chromatography and tandem mass spectrometry. The deuterated internal standard, M6G‐d3, was mixed with the prepared samples at 10 ng/mL as the final concentration. The samples were transferred into the C 18 solid‐phase extraction columns with gradient elution for solid‐phase extraction. The mobile phase consisted of methanol and 0.05% formic acid (pH 3.2). Multiple reaction monitoring has been applied to analyze gemfibrozil ( m / z 249.0 → 121.0) in anion mode, and M6G‐d3 ( m / z 465.1 → 289.1), morphine ( m / z 286.0 → 200.9), and M3G and M6G ( m / z 462.1 → 286.1) in the positive ion mode. The method has a linear calibration range from 0.05 to 10 ng for gemfibrozil, morphine, and M3G and M6G with correlation coefficients >0.993. The lower limit of quantitation for all four analytes was 0.05 ng/mL, relative standard deviation of intra‐ and interday precision was less than 10.5%, and the relative error of accuracy was from −8.2 to 8.3% at low, medium, and high concentrations for all the analytes. In conclusion, gemfibrozil can influence the morphine antinociception after coronary heart disease induced chronic angina by the change in one of morphine metabolites', M3G, distribution in mouse brain.