HPLC-MS测定人血浆中非那雄胺及相对生物利用度

Determination of Finasteride in Human Plasma by HPLC-MS and Study on the Bioequivalence of Domestic and Import Finasteride Tablets

目的:建立正常人血浆中非那雄胺的 HPLC-MS 测定法,研究2种非那雄胺片在正常人的相对生物利用度。方法:以普罗帕酮为内标,血浆样品沉淀蛋白后,经 Nucleodur C_(18)柱分离后采用质谱检测器检测,20名健康志愿者采用自身对照随机交叉试验设计,分别单剂量口服非那雄胺片5mg 后测定两者相对生物利用度。结果:非那雄胺与内标分离度好,内源性杂质不干扰测定,回归方程为 Y=5.930×10^(-3)X+2.363×10^(-3),r=0.9998。在1.53～306.7ng·mL^(-1)。范围内非那雄胺浓度与峰面积比的线性关系良好,定量限为1.53ng·mL^(-1),回收率为95.8%～103.6%(n=5),日内 RSD 为2.2%～4.7%(n=5);日间 RSD 为2.2%～3.6%(n=5)。单次服用5mg 非那雄胺片试验制剂或参比制剂后的药代动力学参数 AUC_(0→24)分别为(359.14±55.78)h·ng·ml^(-1)和(351.42±57.84)h·ng·mL^(-1),AUC_(0→∞)分别为(378.74±59.28)h·ng·mL^(-1)和(371.10±59.72)h·ng·mL^(-1),C_(max)分别为(46.80±6.29)ng·mL^(-1)和(48.63±8.47)ng·mL^(-1),T_(max)分别为(1.90±0.50)h和(1.80±0.41)h。相对生物利用度为103.20%±15.08%。结论:该方法简单,准确度高,灵敏度好,可用于非那雄胺在人体内过程研究。方差分析结果表明2种制剂的主要药动学参数之间无明显差异,双单侧 t 检验结果表明两种制剂为生物等效制剂。

Objective ： An HPLC - MS method for the determination of finasteride in plasma was established and the bioequivalence of domestic and imported finasteride tablets was studied. Method： Samples were pre - treated by acetonitrile and analyzed by a Nucleodur C18 column. The HPLC - MS was carried out by a mobile phase consisting of CH3CN - 10 mmol· L^-1CH3COONH4 （65： 35） ,the flow rate was 0. 85 mL · min^-1 ,and the internal standard was propafenone. A single oral dose of 5 mg domestic and imported finasteride tablets was administered to 20 healthy volunteers in a randomized crossover study. Finasteride concentration in plasma was determined by the newly developed HPLC - MS method, and the pharmacokinetics and bioavailability were studied. Results： The calibration curve was linear with the range 1.53 to 306. 7 ng · mL^- 1 （ r = 0. 9998 ）, the recovery of finasteride from plasma was in the range of 95.8 - 103.6% （n =5） ,inter -day and intra-day RSD were 2.2% -4.7% （n =5）and 2.2% - 3.6% （ n = 5 ） respectively. The AUC0→24 of the two tablets were （ 359. 14 ± 55.78 ） h·ng· ml^-1 and （ 351.42 ± 57.84）ng·h·mL^-1,AUC0→∞were（378. 74 ±59.28）h · ng· mL^-1 and（371.10 ±59.72）h · ng · mL^-1,Cmqx were （46.80 ± 6. 29 ） ng · mL^-1 and （48.63 ± 8.47 ） ng · mL^- 1, Tmax were （ 1.90 ± 0. 50 ） h and （ 1.80 ± 0. 41 ） h, the relative bioavailability of domestic to imported tablets was 103.20% ± 15.08%, respectively. Conclusion： Results of statistics show there were no significant differences between the two preparations, and bioequivalence is observed. The established HPLC - MS method is accuracy and sensible for finasteride determination.