HPLC-MS/MS测定人血浆中阿德福韦及阿德福韦酯胶囊的相对生物利用度研究

Pharmacokinetics and bioequivalence of adefovir dipivoxil capsules in human plasma by HPLC-MS/MS method

目的建立人血浆中阿德福韦的HPLC-MS/MS测定法,研究阿德福韦酯胶囊与普通片剂在中国健康男性志愿者的相对生物利用度,评价两者的生物等效性。方法以阿昔洛韦为内标,血浆样品经甲醇蛋白沉淀,HederaODS-2 C18柱（150 mm×2.1 mm,5μm）分离后,以HPLC-MS/MS法检测。18名健康男性志愿者采用双周期随机交叉试验设计,分别单剂量口服阿德福韦酯胶囊与普通片剂10 mg后于不同时间点取血,以建立的方法测定血浆中阿德福韦的浓度,计算药动学参数,评价两制剂的生物等效性。结果阿德福韦与内标分离良好,内源性杂质不干扰测定,在0.5-100 ng.mL-1（r=0.998 6）阿德福韦浓度与峰面积比的线性关系良好,定量下限为0.5 ng.mL^-1,萃取回收率为71.2%-72.8%（n=5）,日内精密度RSD为4.3%-7.7%（n=5）,日间精密度RSD为2.9%-8.0%（n=15）。单次口服10 mg阿德福韦酯胶囊和普通片剂后的AUC0-t分别为（416.0±106.8）和（410.8±105.9）ng.h.mL^-1;AUC0-∞分别为（425.5±107.1）和（422.1±106.5）ng.h.mL-1;Cmax分别为（35.23±9.6）和（34.31±8.6）ng.mL-1;tmax分别为（1.7±0.5）和（1.6±0.8）h;t1/2分别为（8.1±1.2）和（8.5±1.3）h。与普通片剂相比,阿德福韦酯胶囊的相对生物利用度为（103.7±24.5）%。结论该方法准确度高,灵敏度好,可用于阿德福韦酯胶囊体内过程的研究。两制剂的体内过程无显著性差别,为生物等效性制剂。

Objective To establish an HPLC-MS/MS method for the determination of adefovir in the plasma,and to evaluate the bioavailability of adefovir dipivoxil capsules and tablets in Chinese healthy male volunteers.Methods With aciclovir as the internal standard,the plasma samples were processed with methanol precipitation and analyzed by HPLC-MS/MS on a Hedera ODS-2 C18（150 mm×2.1 mm,5 μm）.A cross over bioequivalence trial was designed and 18 Chinese healthy male volunteers were divided into 2 groups.Each group was orally given adefovir dipivoxil capsules or tablets 10 mg.The pharmacokinetic parameters and relative bioavailability were calculated to evaluate the bioequivalence of the two preparations.Results The calibration curve was linear within 0.5-100 ng·mL-1（r=0.998 6）.The extraction recovery of adefovir was 71.2%-72.8%（n=5）,and the intra-day and inter-day RSD was 4.3%-7.7%（n=5） and 2.9%-8.0%（n=15）,respectively.The AUC0～t of capsules and tablets was（416.0±106.8） and（410.8±105.9） ng·h·mL-1,AUC0～∞（425.5±107.1） and（422.1±106.5） ng·h·mL-1,Cmax（35.23±9.6） and（34.31±8.6） ng·mL-1,tmax（1.7±0.5） and（1.6±0.8） h,and t1/2（8.1±1.2） and（8.5±1.3） h.The relative bioavailability of adefovir dipivoxil capsules to the reference tablets was（103.7±24.5）%.Conclusion The established HPLC-MS/MS method is accurate and sensible.It can be used to study the pharmacokinetics of adefovir in human plasma.The two preparations are bioequivalent.