柱前衍生-固相萃取高效液相色谱法快速测定丙戊酸血药浓度

Rapid Determination of Valproic Acid in Serum by Precolumn Derivation High Performance Liquid Chromatography with Solid Phase Extraction

目的:建立柱前衍生-固相萃取高效液相色谱法测定丙戊酸血药浓度分析方法,去除过量未反应的衍生试剂。方法:选择2-溴-对硝基苯乙酮为衍生试剂,环己烷羧酸为内标,采用 Accubond SPE C_(18)小柱处理反应液,Nova-pak C_(18)柱(4μm,3.9mm×150mm)为分析柱;甲醇-水(77:23)为流动相,检测波长为262nm,流速1 mL·min^(-1),柱温25℃。结果:反应液中90%以上未反应的衍生试剂被去除,内标及丙戊酸衍生物被保留和纯化。内标环己烷羧酸和丙戊酸的保留时间分别为3.0和4.5min,线性范围为16.6～265μg·mL^(-1),相关系数为0.9998。平均回收率为99.85%,日内日间误差 RSD 均小于5.16%。结论:本法快速简便、结果准确,非常适合临床常规监测需要。

objective： To establish a rapid analysis method for the determination of valproic acid in serum by precolumn derivation high performance liquid chromatography with solid phase extraction. Superfluous unreacted derivative reagent was removed from reaction solution. Methods ：2 -Bromo -4＇-nitroacetophenone was selected as derivative reagent. Cyclohexanecarboxylic acid was used as an internal standard. Internal standard and valproic acid derivation were extracted from reaction solution with accubond SPE C18 column. The analytical column was Nova -pak C18 column （3.9 mm × 150 mm,4μm）. The mobile phase consisted of methanol- water（77： 23）. The detection wavelength was 262 nm and the flow rate was 1 mL · min^- 1, column temperature was 25℃. Results： More than 90% unreacted derivation reagent was removed from reaction solution, internal standard and valproic acid derivation were retained and purified. The retaintion time of cyclohexanecarboxylic acid and valproic acid was 3.0 and 4. 5 min, respectively. The linear range of valproic acid was 16. 6 -265μg·mL^-1 and correlation coefficient was 0. 9998. The relative recoveries of valproic acid was 99. 85%. The relative standard deviation（RSD） of within day and between day were all less than 5. 16%. Conclusion： ：This method is found to be rapid, accurate, sensitive and more suitable for clinical practice application.