RP-HPLC测定人血浆中头孢特仑浓度及人体生物等效性研究

Determination of Cefteram in Human Plasma by HPLC and Its Bioequivalence Study

目的建立头孢特仑血浓度反相高效液相色谱测定方法,进行头孢特仑新戊酯片的人体生物等效性研究。方法采用单剂两周期交叉试验设计,20名健康男性志愿者随机单剂量po头孢特仑新戊酯片试验制剂或参比制剂100mg,按设定时间采集肘静脉血,采用Luna C18(2)(4.6mm×250mm,5μm)色谱柱,以甲醇-0.6%冰醋酸(三乙胺调pH至4.0)(30:70)为流动相,流速1mL.min-1,检测波长260nm,测定头孢特仑血浓度计算其药动学参数,评价两制剂的生物等效性。结果本方法最低定量限为0.025mg.L-1,在0.05～3.2mg.L-1(r=0.9989)内线性关系良好,高、中、低浓度批内、批间RSD均小于5.8%。结论本试验所建立的头孢特仑测定方法准确、灵敏、快速,适用于头孢特仑新戊酯片人体药动学研究。头孢特仑新戊酯2制剂主要药动学参数符合生物等效的假设,为生物等效制剂。

OBJECTIVE To establish a HPLC method for the determination of cefteram plasma concentrations in 20 healthy volunteers and to study its bioequivalence. METHODS A single dose of 100 mg cefteram pivoxil test or reference tablets was given to 20 healthy volunteers in a two-periods cross-over design. Plasma samples were collected and quantitated by RP-HPLC method using Luna C18 （2） （4. 6 mm× 150 mm, 5 um） column and mobile phase of methanol- （0. 6% ）glacial acetic acid （pH adjusted to 4. 0 with triethylamine） at flow rate of 1.0mL·min^-1, and detection wavelength of 260 nm. The pharmacokinetic parameters and relative bioavailability were calculated using DAS2.0. RESULTS The limit of detection for cefteram was 0. 05 mg · L^-1 , and the excellent linearity obtained in the range of 0. 05 -3.2mg · L^-1 （r = 0. 998 9）. The relative standard deviations of intra-batch and inter- batch determination were less than 5.8%. CONCLUSION The established method is sensitive, fast and accurate, and can be successfully used for the determination of cefleram in human plasma and for cefteram pivoxil pharmacokinetic studies. The statistic analysis showed that the two preparations were bioequivalent.