目的建立同时测定人血浆中抗癫痫药物磷苯妥英(FOS)、苯妥英(PHT)及其主要代谢产物4′-羟苯妥英(4′-HPPH)浓度的高效液相色谱法。方法以盐酸普萘洛尔为内标,血浆样品100μL经20%磷酸酸化后以乙酸乙酯1 mL萃取,提取后氮气吹干,加入流动相100μL复溶,取20μL进样测定。色谱柱采用Agilent RX-C₈柱(4.6 mm×250 mm,5μm),柱温:40℃;流动相为甲醇-乙腈-0.1%三氟乙酸水溶液(23∶17∶60),流速为1.5 mL·min^-1,紫外检测波长210nm。结果血浆内源性杂质和常用合并用药对样品测定无干扰,色谱峰分离良好。FOS、PHT、4′-HPPH的线性范围分别是1～400,0.5～50和0.5～10 mg·L^-1,相关系数(r)均大于0.999 1。平均方法回收率为92.79%～107.90%。各组分的日内、日间RSD均小于8%。结论该方法具有良好的准确性、精密性和灵敏性,且操作快速、简便,各组分之间分离良好,适用于临床血药浓度的监测。

OBJECTIVE To develop a simple reversed-phase high-performance liquid chromatography(HPLC) method for thesimultaneous determination of the antiepileptic drugs(AEDs) phenytoin(PHT),its prodrug fosphenytoin(FOS),and its metabolite 4′-hydroxyphenytoin(4′-HPPH) in human plasma.METHODS Plasma sample(100 μL) pre-treatment involved acidification with 20% phosphoric acid,simple extraction with 1 mL of acetic ester containing 10 mg·L^-1 propranolol hydrochloride as internal standard and evaporation of solvent,followed by injection of reconstituted sample onto a Agilent RX-C₈ column(4.6 mm × 250 mm,5 μm).The mobile phase consisted of methanol-acetonitrile-0.1% triflouroacetic acid(23∶17∶60),and delivered at a 1.5 mL·min^-1.The analytes were detected at the wavelength of 210 nm.RESULTS The method was found to be linear over the concentration ranges investigated,1-400 mg·L^-1 for FOS,0.5-50 mg·L^-1 for PHT and 0.5-10 mg·L^-1 for 4′-HPPH,respectively.The intra-and inter-day reproducibility was good with the RSD of 8% or below and relative recovery was ranged from 92.79% to 107.90% for all analytes.CONCLUSION This method was proved to be accurate,sensitive and convenient and suitable for clinical therapeutic drug monitoring.