基于2D-HPLC结合捕集柱技术测定人血中双氯芬酸钠浓度及其应用

Determination of Diclofenac Sodium in Human Plasma Based on 2D-HPLC Coupled with Trapping Column and Its Application

目的建立二维高效液相色谱法(2D-HPLC)结合捕集柱技术测定健康受试者血中双氯芬酸钠的浓度,并将其应用到药代动力学和生物等效性研究。方法采用随机交叉试验设计,18例健康男性志愿者分别高脂饮食后口服双氯芬酸钠缓释片受试制剂或参比制剂,2D-HPLC测定血浆中双氯芬酸钠的血药浓度,计算药动学参数及相对生物利用度。结果 18名受试者单次口服100 mg双氯芬酸钠缓释片受试制剂或参比制剂后的主要药动学参数AUC0→16、AUC0→∞、tmax、ρmax分别为(2 591.6±705.8)和(2 588.8±772.0)ng·h·m L^(-1)、(2 896.4±839.7)和(2 700.3±806.1)ng·h·m L^(-1)、(4.6±0.7)和(4.4±0.9)h、(1 332.8±912.5)和(1 325.7±706.3)ng·m L^(-1)。受试制剂对参比制剂的相对生物利用度为(102.4±15.1)%。结论2D-HPLC结合捕集柱技术测定人血浆中双氯芬酸钠浓度简单、快速、专属性高,高脂饮食后单次口服双氯芬酸钠缓释片受试制剂和参比制剂生物等效。

OBJECTIVE To establish a method for determination of diclofenac sodium in human plasma under high-fat meal condition using two-dimensional liquid chromatography（ 2D-HPLC） coupled with trapping column,and to evaluate the bioequivalence and the bioavailability of diclofenac sodium sustain-released tablets in healthy volunteers. METHODS Under fed state,eighteen healthy male volunteers were divided into two groups by an open,randomized two period crossover design with a single dose of diclofenac sodium sustain-released tablets. The plasma concentrations were determined by 2D-HPLC method,and calculated pharmacokinetic parameters and bioavailability. RESULTS The main pharmacokinetic parameters of test and reference preparations after a single dose were：AUC0→16was（ 2 591. 6 ± 705. 8） and（ 2 588. 8 ± 772. 0） ng·h·m L^-1; AUC0→∞was（ 2 896. 4 ± 839. 7） and（ 2 700. 3 ± 806. 1）ng·h·m L^-1; tmaxwas（ 4. 6 ± 0. 7） and（ 4. 4 ± 0. 9） h; ρmaxwas（ 1 332. 8 ± 912. 5） and（ 1 325. 7 ± 706. 3） ng·m L^-1,respectively. The relative bioavailability of test preparation in single dose was（ 102. 4 ± 15. 1） %. CONCLUSION The 2D-HPLC method coupled with trapping column is a simple,rapid and specific for determination of diclofenac sodium in human plasma. The results of statistical analysis indicate that the two preparations are bioequivalent in healthy volunteers with a single dose under high-fat fed condition.