高效液相色谱法测定阿仑膦酸钠的尿药浓度

HPLC determination of alendronate sodium in human urine

目的建立阿仑膦酸钠尿药浓度的高效液相色谱-荧光检测法。方法采用Shiseido Capcell Pak C18(150 mm×4.6 mm,5μm)色谱柱;梯度洗脱,流动相0.01 mol.L-1磷酸氢二钠(A)-乙腈(B);流速为1.0 mL.min-1;柱温35℃;内标物:帕米膦酸钠;荧光检测:激发波长为260 nm,发射波长为310 nm。结果阿仑膦酸钠峰与内标物峰面积比值在20～1 000μg.L-1呈良好的线性关系(r=0.996 1)。方法准确度与日内及日间RSD均符合要求,定量下限为20μg.L-1,提取回收率稳定,成功应用于4名男性健康受试者体内阿仑膦酸钠尿药浓度的测定。结论该法为临床研究阿仑膦酸钠体内尿药浓度提供了方法。

Objective To determine the alendronate sodium concentration in human urine by HPLC with fluorescence detection.Methods Pamidronate sodium was used as the internal standard,and the liquid chromatography was performed on a Capcell Pak C18 column（4.6 mm×150 mm,5 μm） with the mobile phase consisiting of acetonitrile/disodium hydrogen phosphate（14∶86,v/v） at 1.0 mL·min-1 at 35 ℃.The total running time was 30 min.The fluorometric detector was operated at 260 nm（excitation） and 310 nm（emission）.The proposed method was applied to the determination of alendronate in the urine samples from 4 volunteers who were given 70 mg alendronate.Results The linearity of calibrated curves of the urine samples was in 20-1 000 μg·L-1（r=0.996 1）.The intra-and inter-day precision expressed as the relative standard deviation was less than 15%.The recovery of alendronate sodium and pamidronate sodium was above 85%.The limit of quantification was 20 μg·L-1of alendronate sodium using 5 mL urine,and the mean excretion rate-time profile of alendronate sodium was presented.Conclusion The method is feasible and reproducible,which offers a satisfactory tool for the pharmacokinetics and relative bioavailability of alendronate sodium in the urine.