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本文建立了RH-HPLC 法测定人尿中苯妥因(PHT)及其代谢物4′—羟基苯妥因(HPPH)。用乙醚为提取溶剂,5-对甲苯基-5-苯基乙内酰脲(MPPH)为内标。色谱分析用 Bil-Sll(?)ODS 柱,含40%乙腈的水溶液为流动相。PHT 和 HPPH 的线性测定范围为0.05—2.0μg/ml 和1.0—100μg/ml,相关系数均优于0.99。平均回收率分别为98.9%和99.9%,平均变异系数分别为4.8%和4.7%。应用本法研究了88名健康志愿者 PHT 的遗传代谢多态性。
This article established a method for the determination of phenytoin (PHT) and its metabolite 4’-hydroxyphenytoin (HPPH) in human urine by RH-HPLC. Using diethyl ether as the extraction solvent, 5-p-tolyl-5-phenylhydantoin (MPPH) was used as an internal standard. Chromatography using Bil-Sll (?) ODS column, containing 40% acetonitrile in water as the mobile phase. The linear range of PHT and HPPH was 0.05-2.0μg / ml and 1.0-100μg / ml, the correlation coefficient was better than 0.99. The average recoveries were 98.9% and 99.9%, respectively. The average coefficients of variation were 4.8% and 4.7% respectively. This method was used to study the genetic polymorphism of PHT in 88 healthy volunteers.