人血浆中瑞格列奈HPLC法测定及其药代动力学研究

Determination of Repaglinide in Plasma by HPLC and Its Pharmacokinetics in Men

目的 :建立了人血浆中瑞格列奈浓度的 HPLC测定方法 ,研究瑞格列奈在人体中药物动力学行为。方法 :血浆样品经酸化后 ,用醋酸乙酯提取 ,色谱柱为 C1 8Shim-pack CLC-ODS,流动相为 0 .1 0 mol/ L柠檬酸 -醋酸钠缓冲液 ( p H4 .0 ) -甲醇 ( 2 7∶ 73) ( v/ v)。紫外检测波长 2 4 3nm。测定了 2 2名受试者单剂量口服瑞格列奈 4mg后血药浓度 -时间过程。结果 :最低检测浓度 2 .5ng/ ml,回收率大于 90 % ,日间和日内的变异系数小于1 5% ,线性范围为 2 .5～ 1 0 0 .0 ng/ ml,符合生物样品分析的要求。受试者口服瑞格列奈片 4 mg后 ,估算的末端相半衰期 0 .83± 0 .31 h,峰时间 0 .75± 0 .4 1 h,峰浓度 53.32± 2 4 .94 ng/ ml,MRT1 .58± 0 .4 1 h,AUC40 74 .95± 30 .57ng· h/ ml。结论 :建立的 HPLC方法适合于瑞格列奈临床研究。

AIM: To establish a reversed phase HPLC method for determination of repaglinide and to study pharmacokentics of repaglinide in men. METHODS: After acidified, the drug was extracted from plasma with acetate ether. Shim pack CLC ODS C 18 column was used. Mobile phase consisted of 27% 0.10 mol/L citrate acid sodium acetate buffer(pH=4.0) and 73% methanol. The eluted peaks were detected by UV detector at 243 nm. RESULTS: The recoveries of repaglinide from plasma were larger than 90% and RSD% of intra day and inter day were smaller than 15%. Calibration curves were linear over 2.5～100.0 ng/ml( r =0.9992) and the minimum detection concentration was 2.5 ng/ml. The pharmcokinetics of repaglinide in 22 healthy subjects was studied. After oral administration of 4 mg repaglinide tablet, the pharmacokinetic parameters were estimated as follows: T 1/2 , 0.83±0.31 h, T max , 0.75±0.41h, C max , 53.32±24.94 ng/ml, MRT 1.58±0.41 h, and, 74.95±30.57 ng·h/ml. CONCLUSION: A reliable and rugged simultaneous HPLC assay for repaglinide was developed. The assay method was convenient for clinical laboratory.