反相高效液相色谱法测定丙戊酸的血药浓度

RP-HPLC determination of valproate concentration in serum

目的建立反相高效液相色谱法测定人血清中丙戊酸的药物浓度。方法以symmetryshield RP18柱(5.0μm,3.9 mm×150 mm)为色谱柱。流动相为甲醇:水(80:20),流速1.0 m//min。Waters 2487双波长紫外检测器,检测波长248 nm。血清用正己烷提取,经α-溴苯乙酮衍生后浓集进样.以环己烷羧酸为内标物。结果线性范围为5.0～250.0μg/ml,r=0.9994;平均回收率为98.23%,高、中、低浓度样品的日间和日内测定相对标准偏差(RSD)均小于3%。结论本法适用于丙戊酸的血药浓度测定。

Objective To establish an RP-HPLC method for determining serum level of valproate. Methods A symmetryshield RP18 column （ 5μm, 3.9mm × 150mm ） was used. The mobile phase, methanol-water （ 80 ：20 ） was used at a flow rate of 1.0ml/min. The detector was Waters 2487 dual λ absorbance detector, which was set at the wavelength of 248nm. Serum sample was extracted with 2ml acetic n-hexane, then valproic acid in organic layer was derived by adding into a-bromoacetophenone, and cyclohexanecarboxylic acid was selected as an internal standard. Results The calibration curve was linear in the range of 5.0 to 250.0μg/ml. The average recovery rate was 98.23 %, and the RSD within-day and between-day were both less than 3 %. Conclusion This method is suitable for therapeutic drug monitoring of phenytoin.