摘要
目的建立简便、快速、灵敏的HPLC-MS法测定人血浆中氟康唑浓度,评价受试和参比制剂的人体生物等效性。方法以甲硝唑为内标,血浆样品用甲醇直接沉淀去蛋白后取上清液进行HPLC-MS分析。色谱柱为LichrospherC18(250mm×4.6mm,5μm),流动相为甲醇-水(65∶35,含40mol/L醋酸铵和0.05%甲酸),流速为0.8mL/min;质谱采用电喷雾离子化方式,选择性离子检测,检测对象为氟康唑的[M+H]+离子(m/z307.2)和内标甲硝唑的[M+H]+离子(m/z172.3),裂解电压为70V。20名健康志愿者随机双交叉口服受试制剂和参比制剂150mg,定时测定人血浆中氟康唑浓度,进行药动学研究,评价生物等效性。结果氟康唑血药浓度在30.69μg/L^20.46mg/L范围内线性关系良好(r=0.9998,n=5),最低检测限为10μg/L。各实验浓度样品检测的精密度良好,日内和日间RSDs均小于10%,提取回收率大于90%。以AUC0-120计算的受试制剂相对生物利用度为(106.4±11.8)%,受试与参比制剂生物等效。结论该方法简便、快速、专属性强、灵敏度高、准确性好,适用于氟康唑血药浓度测定及药动学研究。
Objective： To establish a HPLC-MS method for rapid determination of fluconazole in human plasma and to assess the pharmacokinetics and bioequivalence of fluconazole capsules in healthy volunteers. Methods： After being deproteined by methanol, plasma samples were separated by HPLC on a reversed-phase lichrospher ClS column （250 mm × 4.6 mm,5 gm） with a mobile phase of methanol-water （65：35, containing 0.05% formic acid and 40 mol/L ammonium acetate） at a flow rate of 0.8mL/min and metronidazole was used as internal standard. HPLC-MS was performed in the selected ion monitoring （SIM） mode using target ions, [M ＋ H]^＋ （m/z： 307.2） for fluconazole and [M ＋ H]^＋ （ m/z： 172.3） for the internal standard, and fragment voltage of 70 V. A randomized crossover trial was performed in 20 healthy volunteers, who took a single 150 mg dose of test capsules or reference capsules respectively, followed by periodically determining fluconazole in human plasma. Results： The standard curves were linear in the range of 30.69μg/L～20.46 mg/L （r = 0.9998, rt = 5） and the low limit of detection for fluconazole was about 10μg/L. The RSDs of intra- and inter-day were all less than 10% and the extraction recovery rates were more than 90% for all test concentrations of samples. The relative bioavailability of test capsules was （106.4±11.8）% and the two preparations were bioequivalent. Conclusion： The assay is proved to be rapid, sensitive, accurate, convenient and specific, and is suitable for the determination of fluconazole in human plasma and pharmacokinetic study.
出处
《药学进展》
CAS
2006年第3期122-126,共5页
Progress in Pharmaceutical Sciences
