固相萃取结合HPLC-MS测定人血浆中佐米曲坦及相对生物利用度

 目的建立人血浆中佐米曲坦的HPLC-MS测定法,研究两种佐米曲坦片的相对生物利用度。方法以舒马曲坦为内标,血浆样品由固相萃取,经Nucleodur C₁₈柱分离后采用质谱检测器检测。18名健康志愿者采用自身对照随机交叉试验设计,分别单剂量口服佐米曲坦片5mg后测定二者相对生物利用度。结果佐米曲坦与内标分离度好,内源性杂质不干扰测定,曲线方程为Y=0.004764X+0.001114,r=0.9995。在0.25～40μg·L^-1内佐米曲坦质量浓度与峰面积比的线性关系良好,最低定量限为0.25μg·L^-1,佐米曲坦萃取回收率为86.7%～96.4%(n=5),内标萃取回收率为79.7%～81.4%(n=15),方法回收率为102.2%～106.0%(n=5),日内精密度(RSD)2.28%～3.42%(n=5);日间精密度(RSD)为3.42%～5.45%(n=5)。单次服用5mg佐米曲坦片试验制剂或参比制剂后的药动学参数AUC_0～16分别为(59.6±24.4)和(56.6±31.2)μg·h·L^-1,AUC_0～∞分别为(62.3±25.9)和(60.2±24.0)μg·h·L^-1,ρ_max分别为(12.4±4.2)和(12.7±4.4)μg·L^-1,t_max(1.9±0.5)和(2.1±0.4)h。相对生物利用度为(107.1±25.7)%。结论该方法简单,准确度高,灵敏度好,可用于佐米曲坦在人体内过程研究。方差分析结果表明,两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。

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

OBJECTIVE To develop a HPLC-MS method for the determination of zolmitriptan in plasma and evaluate the bioequivalence of domestic and imported zolmitriptan tablets.METHODS Samples were pre-treated by solid extraction and analyzed by a Nucleodur C₁₈ column. The mobile phase consisted of CH₃ CN-CHCOOH(0.05%) = 10∶90, and the internal standard was sumatriptan. A single oral dose of 5 mg domestic and imported zolmitriptan was administered to 18 healthy volunteers in a randomized crossover study. Zolmitriptan concentration in plasma was determined by the developed LC-MS method, and the pharmacokinetics and bioavailability were studied. RESULTS The calibration curve was linear over the range of 0.25～40 μg·L^-1(r=0.999 5), the solid extraction recoveries of zolmitriptan and sumatriptan respectively were between 86.7%～96.4%(n=5)and 79.7%～81.4%(n=15), the method recovery was in the range of 102.2%～106.0% (n=5) , inter-day and intra-day RSDs were 2.3%～3.4%(n=5) and 3.4%～5.4%(n=5),respectively. The AUC_0～t of the two tablets were(59.6±24.4) and (56.6±31.2) μg·h·L^-1 ,AUC_0～∞ were (62.3±25.9) and (60.2±24.0) μg·h·L^-1,ρ_max were(12.4±4.2) and(12.7±4.4) μg·L^-1,t_max were (1.9±0.5) and (2.1±0.4) h, the relative bioavailability of domestic to imported tablet was (107.1±25.7)%. CONCLUSION Statistice analysis shows no significant differences between the two preparations, and bioequivalence is observed. The established HPLC-MS method is accuracy and sensible for zolmitriptan determination.