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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×0.25μm,0.25 mm),采用程序升温,分流进样,血浆经磷酸二氢钠缓冲液(p H 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·m L-1,检出限分别为1.6和1.5 ng·m L-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。
OBJECTIVE To establish simultaneous determination of valproic acid (VPA) and its hepatotoxic metabolite 2-propyl-4-ene-VPA in human plasma by gas chromatography-mass spectrometry (GC-MS) Analysis methods for the clinical monitoring of plasma concentration of valproic acid and warning of valproic acid-induced hepatotoxicity to provide detection means. Methods The chromatographic column was HP-5MS quartz capillary column (30 m × 0.25 μm, 0.25 mm). The temperature was programmed and shunted. The plasma was acidified with pH 5 and extracted with ethyl acetate. The samples were derivatized with derivatization reagent (BSTFA + 1% TMCS) and quantified by full-scan (SCAN) qualitatively using a selective ion monitoring (SIM) internal standard method. Results The results showed that valproic acid and 2-propyl-4-pentenoic acid were separated well under this chromatographic conditions with a good linearity (r> 0.999). The recovery rates of both were above 96% and the intra-day and inter-day precision were less than 2.49 % And 4.63% respectively. The minimum limits of quantification were 5.21 and 5.25 ng · m L-1, respectively. The limits of detection were 1.6 and 1.5 ng · m L-1, respectively. Conclusion The method of simultaneous determination of valproic acid and 2-propyl-4-pentenoic acid in human plasma by phase chromatography-mass spectrometry is of good accuracy and precision. The method has high extraction recovery and is suitable for the clinical application of valproic acid Blood concentration monitoring and warning of valproic acid hepatotoxicity detection means. In this study, the content of valproic acid and 2-propyl-4-pentenoic acid in Chinese people were determined by phase-HPLC-MS simultaneously for the first time.