【摘要】 目的建立测定人血浆中伏立康唑的高效液相色谱-串联质谱法(HPLC-MS/MS),研究2种伏立康唑片的相对生物利用度。方法血浆样品经酸化后用甲醇沉淀蛋白,经Waters Atlantis C18柱分离,流动相为乙腈-0.2%冰醋酸水溶液(内含20 mmol.L-1醋酸铵)(60∶40,v/v),流速为0.2 mL.min-1;选择多反应离子检测(MRM)方式进行定量分析,用于监测的离子为质荷比m/z350.9→281.4(伏立康唑)和m/z307.9→220.3(内标氟康唑)。18名健康志愿者以随机交叉方式分别单次口服伏立康唑分散片T或伏立康唑片R 0.2 g后于不同时间点取血,样品以新建立的HPLC-MS/MS法测定,研究比较两制剂的药动学及相对生物利用度。结果伏立康唑与血浆中内源性杂质分离度好,伏立康唑浓度在27.35~3 500 ng.mL-1与峰面积比线性良好,最低定量浓度为27.35ng.mL-1。蛋白沉淀绝对回收率为86.2%~88.8%,相对回收率为97.9%~105.7%,日内精密度(RSD)<9.8%,日间精密度(RSD)<9.4%。单剂量口服伏立康唑分散片T和伏立康唑片R 0.2 g后2种制剂的Cmax为(717.9±325.1)、(671.8±243.3)μg.L-1;tmax为(1.1±0.5)、(1.1±0.4)h;AUC0~24为(3 729.1±1 887.7)、(3 811.2±1 836.6)μg.h.L-1;t1/2为(6.7±1.9)、(6.7±1.7)h。与R相比,T制剂相对生物利用度为(97.3±13.0)%。结论该方法简单快速,灵敏度高,可用于伏立康唑的体内过程研究。方差分析表明伏立康唑分散片T与伏立康唑片R中伏立康唑的主要药动学参数之间均无明显差异,双单侧t检验结果表明两制剂为生物等效制剂。更多还原

【Abstract】 Objective To develop an HPLC-MS/MS method for the determination of voriconazole in human plasma and evaluate the bioequivalence of voriconazole tablets.Methods After acidization and protein precipitation with methanol,analysis was performed on a Waters HPLC system using a Waters Atlantis C18 column and isocratic elution with the mobile phase of acetonitrile-0.2% glacial acetic acid(contain 20 mmol·L-1 ammonium acetate)(60∶40,v/v) at 0.2 mL·min-1,and a tandem mass spectrometer Micromass,equipped with an electrospray ionization interface,operated in a positive mode.The multiple reaction monitor(MRM) was used to detect voriconazole(m/z 350.9→281.4) and fluconazol(m/z 307.9→220.3,as the internal standard).In a randomized crossover design,18 healthy male volunteers were given 0.2 g voriconazole tablet T(test drug) or tablet R(reference drug).The blood samples were obtained and determined by the newly-developed HPLC-MS/MS method and the pharmacokinetics and bioavailability were studied.Results The calibration curve of voriconazole was linear over 27.35~3 500 ng·mL-1 with the limit of quantity 27.35 ng·mL-1.The absolute recoveries were at 86.2%~88.8%,and the relative recoveries at 97.9%~105.7%.Inter-day and intra-day RSD was less than 9.8% and 9.4%,respectively.Pharmacokinetic parameters of voriconazole after a single dose of tablet T and R were as follows:Cmax was(717.9±325.1),(671.8±243.3)μg·L-1,tmax was(1.1±0.5),(1.1±0.4)h,AUC0~24 was(3 729.1±1 887.7),(3 811.2±1 836.6)μg·h·L-1,t1/2 was(6.7±1.9),(6.7±1.7)h,and the relative bioavailability was(97.3±13.0)%.Conclusion The method is selective,sensitive and rapid for voriconazole determination.ANOVA and two one-sided t-tests analysis show that there is no significant difference between the pharmacokinetic parameters of the two formulations and they are bioequivalent.更多还原