RP-HPLC测定巴洛沙星的人体血药浓度及生物等效性研究

Determination of Balofloxacin in Human Plasma by RP-HPLC and Its Application to Bioequivalence Research

目的建立人血浆中巴洛沙星的HPLC测定法,研究巴洛沙星胶囊健康人体相对生物利用度。方法采用双周期自身交叉对照试验设计,20名健康志愿者口服受试巴洛沙星胶囊和参比巴洛沙星片各200 mg后采用HPLC测定血浆药物浓度。色谱柱为Lichmspher ODS(4.6 mm×25 cm,5μm)柱,流动相为水-乙腈-三乙胺-冰醋酸(78:22:1:1),紫外检测波长295 nm。结果巴洛沙星在0.03～4.0 mg·L^(-1)内线性关系良好(r=0.9997),提取回收率为85.37%～88.34%,相对回收率为98.45%～103.63%。受试、参比制剂的主要药动学参数如下:t_(1/2)(8.59±1.48)和(8.79±1.43)h,t_(max) (1.0±0.5)和(1.1±0.5)h,ρ_(max)(2.09±0.76)和(1.95±0.56)mg·L^(-1),AUC_(0-36h)(15.15±2.98)和(16.77±11.36)mg·h·L^(-1),AUC_(0-∞)(16.00±3.13)和(18.64±11.72)mg·h·L^(-1),经对数转换根据两种制剂血药浓度-时间曲线下面积计算国产巴洛沙星胶囊剂相对生物利用度为(99.1±14.0)%。经方差分析及双单侧t检验,显示两制剂无显著性差异,具有生物等效性。结论此法简便、灵敏、准确、稳定,可用于体内药物分析,巴洛沙星胶囊和巴洛沙星片具有生物等效性。

OBJECTIVE To develop a HPLC method for the determination of balofloxacin in plasma and evaluate relative bioavailability of domestic balofloxacin capsule in healthy volunteers. METHODS A single oral dose of 200 mg balofloxacin domestic capsule and tablet was given to 20 health volunteers in a randomized crossover study. The balofloxacin concentrations in plasma were determined by HPLC method. The prepared sample was separated on the column of Lichrospher ODS （4. 6 mm ×25 cm, 5μm）, with the mobile phase consisting of H： O-Acetonitrile-Triethylmnine-HAc （78： 22：1 ： 1 ） . The eluted peaks were detected by UV detector at 295 nm. RESULTS The calibration curve was linear in the range of 0. 03 % -4. 0 mg ·L-1 （r =0. 999 7 ）, the clean-up recovery of balofloxacin was 85.37% - 88.34% and the analytical recovery was 98.45% - 103.63%. The pharmacokinetic parameters of domestic capsules and tablets were as follows respectively ： t1/2 （8.59 ± 1.48 ） and （8. 79 ±1.43 ） h, tmax （ 1. 0 ± 0. 5 ） and （1.1±0.5） h, Pmax （2.09_＋0.76） and （1.95 ±0.56） mg·L-1, AUC0_36h （15. 15 ±2.98） and （16.77 ±11.36） mg· h ·L-1 , AUC0-m （ 16. 00 ±3. 13 ） and （ 18. 64 ±11.72） mg · h ·L-1. The relative bioavailability of domestic capsules was （99. 1 ± 14. 0）%. The domestic capsules and tablets were bioequivalent. CONCLUSION The method was simple, sensitive, reliable and good enough to be used in pharmacokinetic study of balofloxacin. Two formulations in research are bioequivalent.