优化LC-MS/MS法测定人血浆非那雄胺浓度

Determination of finasteride in healthy volunteers by optimized liquid chromatography-tandem spectrometry

目的建立适用于血浆非那雄胺浓度检测的优化液相色谱-串联质谱(LC-MS/MS)检测方法,评价非那雄胺片剂的药动学参数。方法20例健康受试者单剂量口服非那雄胺受试制剂和参比制剂(均为5 mg),以曲安奈得为内标,血浆样品经液-液萃取后,采用C_(18)(100 mm×2.1 mm,5μm)柱,以乙腈-水=65:35(含甲酸铵5 mmol/L)为流动相。采用电喷雾离子源(ESI)、多反应离子监测(MRM)和正离子扫描测定非那雄胺的浓度,计算主要药动学参数及相对生物利用度。结果本试验建立的LC-MS/MS检测方法提示,非那雄胺在0.5～75 ng/mL内线性关系良好,日内、日间精密度均小于7.0%,该方法回收率为86.44%～115.5%。结论经优化的LC-MS/MS方法用于血浆样本非那雄胺检测准确、灵敏,可供临床药动学研究参考。

Objective To establish the optimized liquid chromatography-tandem spectrometry （LC-MS/MS） method for the determination of finasteride in healthy volunteers and to evaluate its pharmacokinetic. Methods Twenty healthy volunteers were administrated finasteride test and reference formulation. Plasma samples were pre-treated by a liquidliquid extraction before injection. An ESI ion source was used and operated in the positive ion mode with multiple reaction monitoring （MRM）. Triamcinolone acetonide was chosen as internal standard and chromatographic separation was achieved on a C18 （100 mm ×2.1mm,5μm）column with a mobile phase consisting of acetonitrile/5 mmol/L ammoniura acetate （65/35, v/v）. Results The method was validated over the concentration range of 0.5-75 ng/mL for finasteride, and showed excellent linearity. The intra- and inter-assay precision was within 7.0% and the assay accuracy was 86.44%- 115.5% over the three concentration levels evaluated. Conclusion Optimized LC-MS/MS method is sensitive, accurate and has been successfully applied in pharmacokinetic study of finasteride tablets in healthy volunteers.