HPLC法测定1例肾功能衰竭患者血浆中更昔洛韦浓度

Determination of ganciclovir in the plasma of a patient with renal failure by HPLC

目的建立人血浆中更昔洛韦的HPLC测定方法,测定肾功能衰竭患者静脉滴注更昔洛韦的血浆药物浓度。方法取血浆样品100 μL,以阿昔洛韦为内标,10%高氯酸沉淀蛋白后,取10 μL进样。HPLC分离采用Shim-pak CLC-ODS(150 mm×60 mm,5 μm)柱,流动相为0.03 mol·mL^-1磷酸二氢铵水溶液∶乙腈=97∶3(磷酸调节pH至3.0),流速为1.0 mL·min^-1,柱温35 ℃,紫外检测波长254 nm。应用上述HPLC方法测定1例肾功能衰竭患者静脉滴注更昔洛韦(75 mg)后的血浆药物浓度。结果标准曲线在0.5~50 μg·mL^-1浓度范围内线性良好(r=0.9999),低、中、高浓度质控样品(1、8、30 μg·mL^-1)的日内、日间精密度(RSD)1.01%~5.55%(n=5),准确度-0.15%~8.00%(n=5),更昔洛韦的提取回收率为98.60%~100.36%。应用该方法成功测定了1例肾功能衰竭患者血浆更昔洛韦药物浓度。结论本方法灵敏度高,准确性好,简便快捷,适用于临床患者血浆中更昔洛韦浓度的测定。

ObjectiveTo develop a high-performance liquid chromatographic(HPLC)method for determination of ganciclovir in the plasma of a patient with renal failure.MethodsAfter addition of acyclovir(internal standard),100 μL plasma samples were prepared based on a simple protein precipitation by adding perchloric acid(10%),and then take 10 μL as sample.The separation was performed on a Shim-pak CLC-ODS(150 mm×60 mm,5 μm)at 35 ℃ using 0.03 mol·mL^-1 ammonium dihydrogen phosphate in aqueous solution/acetonitrile(97∶3)(pH adjusted to 3.0 by H3PO4) as mobile phase.The flow rate was 1.0 mL·min^-1 and the wavelength was 254 nm.The above HPLC method was used to determine the plasma concentration of ganciclovir after intravenous infusion at a dose of 75 mg in a patient with renal failure.ResultsThe calibration curve was linear in the range of 0.5~50 μg·mL^-1(r=0.9999). The intra-run and inter-run RSDs were 1.01%~5.55%(n=5)for the high,middle,and low check samples(1,8,30 μg·mL^-1),and the accuracy was -0.15%~8.00%(n=5). The extraction recoveries were 98.60%~100.36%(n=5). The method was successfully applied to the determination of plasma concentration of ganciclovir in the patient with renal failure. ConclusionThe method is simple,sensitive,and accurate,and it can be used for the measurement of the concentration of ganciclovir in plasma of clinical patients.