目的 建立同时测定人血浆中丙戊酸(VPA)及其肝毒性代谢物2-丙基-4-戊烯酸(4-ene-VPA)含量的气相色谱-质谱(GC-MS)联用定量分析方法,为临床上丙戊酸血药浓度的监测及预警丙戊酸所致肝毒性的发生提供检测手段。方法 色谱柱为HP-5MS石英毛细管柱(30 m×0.25 μm,0.25 mm),采用程序升温,分流进样,血浆经磷酸二氢钠缓冲液(pH 5)酸化后用乙酸乙酯萃取,再用衍生化试剂(BSTFA+1%TMCS)衍生后进样,采用全扫描(SCAN)定性,选择离子监测(SIM)内标法定量。结果 丙戊酸和2-丙基-4-戊烯酸在此色谱条件下分离良好,线性关系良好(r>0.999),萃取回收率均在96%以上,日内日间精密度均分别小于2.49%和4.63%,最低定量下限分别为5.21和5.25 ng·mL-1,检出限分别为1.6和1.5 ng·mL-1。结论 相色谱-质谱可同时测定人血浆中丙戊酸和2-丙基-4-戊烯酸含量,此方法具有良好的准确度与精密性,萃取回收率高,适合作为临床上丙戊酸血药浓度监测及预警丙戊酸肝毒性发生的检测手段。本实验首次采用相色谱-质谱同时测定中国人群中丙戊酸和2-丙基-4-戊烯酸的含量,对于丙戊酸临床安全用药提供依据。
Abstract
OBJECTIVE To establish a method for simultaneous determination of valproic acid(VPA) and 2-propyl-4-pentenoic acid(4-ene-VPA) in human plasma by gas chromatography-mass spectrometry(GC-MS) to provide a means for monitoring the plasma concentration of VPA and early warning the valproic acid induced liver toxicity.METHODS The analytes as TMS derivatives by BSTFA+1%TMCS were separated and identified on HP-5MS column(30 m×0.25 μm,0.25 mm) by temperature-programming. Split injection mode was used. The analytes spiked in plasma were extracted with ethyl acetate after acidification with NaH2PO4 buffer solution(pH 5) and then determined and identified using selected ion monitoring(SIM) mode and scan mode, respectively. The internal standard method was used for the determination.RESULTS VPA and 4-ene-VPA were well separated on their SIM chromatograms and also on the total ion current(TIC) chromatograms. The calibration curves for VPA and 4-ene-VPA showed excellent linearity(r>0.999). Extraction recoveries were higher than 96%. Intra-day and inter-day RSDs were all lower than 2.49% and 4.63%, the lower limit of quantitation is 5.21 and 5.25 ng·mL-1, respectively. The detection limits were 1.6 and 1.5 ng·mL-1, respectively.CONCLUSION An optimized analytical method for simultaneous analysis of VPA and 4-ene-VPA in plasma are developed using GC-MS. With good accuracy and precision, high extraction recoveries, it is suitable as a means for monitoring the plasma concentration of VPA and early warning the valproil acid induced liver toxicity. In this paper, it is the first time of simultaneously determination VPA and 4-ene-VPA among Chinese people by GC-MS for providing basis for VPA clinical application.
关键词
丙戊酸 /
2-丙基-4-戊烯酸 /
肝毒性 /
气质联用
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Key words
valproic acid /
2-propyl-4-pentenoic acid /
hepatotoxicity /
gas chromatography-mass spectrometry(GC-MS)
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中图分类号：
R969.1
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参考文献
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脚注
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基金
湖南省自然科学基金资助项目(11JJ5064)
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