高效液相色谱-质谱联用法快速测定人血浆中丙戊酸钠浓度

目的：建立有效测定人血浆中丙戊酸钠浓度的高效液相色谱-质谱连用（HPLC-MS/MS）的方法。方法：血浆样本经乙腈沉淀蛋白后，以替米沙坦为内标，采用Agilent ZORBAX300SB-C₁₈柱（2.1 mm×150 mm， 5 μm）色谱柱，乙腈-20 mmol·L^-1乙酸胺（55：45）为流动相，流速0.25 mL·min ^-1，以液相色谱分离，电喷雾离子化串联质谱进行检测，采用多反应监测（MRM）测定丙戊酸钠（m/z 143.0→m/z 143.0）和内标替米沙坦（m/z 513.5→ m/z 469.4）的浓度。结果：校准曲线线性浓度范围从0.2~100 μg·mL^-1，最低定量限为0.2 μg·mL^-1。3个不同浓度（低、中、高）丙戊酸钠提取回收率分别为90.46%，93.18%，95.31%。日内和日间精密度均小于10%和8%。结论：此法简单灵敏，重现性好。已成功应用于丙戊酸钠片在中国健康志愿者的药动学研究。

Fast determination of sodium valproate in human plasma by HPLC-MS/MS

OBJECTIVE To establish a high performance liquid chromatography-mass spectrometry (HPLC-MS/MS) method for the determination of sodium valproate in human plasma. METHODS Plasma samples were precipitated with acetonitrile and analyzed with an Agilent ZORBAX300SB-C₁₈ (2.1 mm×150 mm, 5 μm) column using acetonitrile-20 mmol · L^-1 amine acetate (55:45) as the mobile phase at a flow rate of 0.25 mL min^-1. The proteins were separated by liquid chromatography and detected by electrospray ionization tandem mass spectrometry. The concentrations of sodium valproate (m/z 143.0→m/z 143.0) and internal standard telmisartan (m/z 513.5→m/z 469.4) were determined by multiple reaction monitoring (MRM).RESULTS The linear concentration range of the calibration curve was from 0.2 to 100 μg·mL^-1, and the lowest limit of quantitation was 0.2 μg·mL^-1. The recovery rates of sodium valproate extracted at three different concentrations (low, medium and high) were 90.46%,93.18% and 95.31%,respectively. The intra-and inter-day precisions were less than 10% and 8%.CONCLUSION The method is simple, sensitive and reproducible. It has been successfully applied to the pharmacokinetic study of valproate sodium tablets in Chinese healthy volunteers.