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目的:建立超快速液相色谱-串联质谱(UFLC-MS/MS)法测定人血浆中丙戊酸浓度。方法:以双氯芬酸钠为内标,血浆样品用甲醇沉淀蛋白后进样分析。色谱柱为Shim-pack XR-ODS II C18柱(2.0 mm×75 mm,2.2μm),以乙腈和5 mmol·L~-1醋酸铵水溶液为流动相进行梯度洗脱,流速0.3 ml·min~-1。质谱采用多反应监测(MRM)扫描模式,以电喷雾离子源(ESI)在负离子电离模式下进行测定,丙戊酸和双氯芬酸钠的定量分析离子分别为m/z 142.8→142.8和m/z 294.0→249.8。结果:血浆中丙戊酸在8.4~168.0μg·ml-1内线性关系良好(r=0.997 5),日内及日间RSD均小于15%,稳定性考察结果良好。选取已用EMIT法检测的丙戊酸血浆样品30例应用本试验所建方法测定,所得结果与EMIT法测定结果进行比较(独立样本t检验),结果表明本试验所建方法的测定结果与EMIT法测定结果的差别无统计学意义。结论:该方法快速、灵敏、专属性强,可用于人血浆中丙戊酸浓度的测定。
Objective: To establish a method for the determination of valproic acid in human plasma by ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS / MS). METHODS: Diclofenac sodium was used as an internal standard and plasma samples were precipitated with methanol and analyzed by injection. The column was eluted with a gradient of acetonitrile and 5 mmol·L -1 ammonium acetate in water as mobile phase at a flow rate of 0.3 ml · min -1 on a Shim-pack XR-ODS II C18 column (2.0 mm × 75 mm, 2.2 μm) -1. Mass spectrometry was performed in multiple ion monitoring (MRM) scanning mode with negative ionization mode using electrospray ionization (ESI). Quantitative analysis of valproic acid and diclofenac sodium was m / z 142.8 → 142.8 and m / z 294.0, respectively → 249.8. Results: The plasma valproic acid had a good linear relationship in the range of 8.4-168.0 μg · ml-1 (r = 0.997 5). The intra-day and inter-day RSD were all less than 15%. The stability was good. Thirty cases of plasma samples of valproic acid, which had been detected by EMIT, were selected according to the method established in this study. The results were compared with the results of EMIT (t-test), and the results showed that the results of this method were similar to EMIT The difference between the results of the determination of law was not statistically significant. Conclusion: The method is rapid, sensitive and specific and can be used for the determination of valproic acid in human plasma.