LC-MS法同时测定厄贝沙坦氢氯噻嗪片中2组分的血药浓度及其生物等效性研究

目的:建立同时测定人血浆中厄贝沙坦和氢氯噻嗪浓度的方法,并评价2种厄贝沙坦氢氯噻嗪片的生物等效性。方法:以L--替11醋22米酸.9沙。铵坦对缓为1冲8内液名标(健氨,康血水受浆调试样节者品pH随经值机沉至双淀9周法.0)期处=交理3叉后5∶单,6采5剂;用用量液于口-定质服量联相分用同析法剂的进量离行的子测受对定试厄,制色贝剂谱沙或柱坦参为m比/Wz制-at剂e4r2s厄7X.3贝T→e沙r-ra坦1R9氢P31.氯08,,噻氢流嗪氯动片噻相以嗪为考m乙/察z腈-其∶12生m96m物.0o→等l·效性。结果:厄贝沙坦和氢氯噻嗪血药浓度分别在2.00～5000μg·L-(1r=0.9984)和2.00～200μg·L-(1r=0.9988)范围内线性关系良好,最低定量限均为2.00μg·L-1,提取回收率均>90%,日内、日间RSD均<10%,厄贝沙坦、氢氯噻嗪方法回收率分别为96.3%～108.0%、98.2%～100.5%。2种制剂的cmax、AUC0～t、AUC0～∞制剂间和周期间均无显著性差异(P>0.05),对tmax进行非参数检验显示亦无显著性差异(P>0.05)。结论:本法特异性强、快速、准确,所需样本少且前处理简单,能同时对厄贝沙坦和氢氯噻嗪的血药浓度进行定量;2种制剂生物等效。

Simultaneous Determination of Plasma Concentrations of 2 Components of Irbesartan Hydrochlorothiazide Tablet and Bioequivalence Study of It

OBJECTIVE： To develop the simultaneous determination method of irbesartan and hydrochlorothiazide in human plasma, and to evaluate the bioequivalence of 2 kinds of Irbesartan hydrochlorothiazide tablets. METHODS： Telmisartan was used as the internal standard. Plasma sample was extracted with acetonitrile and centrifuged. LC-MS/MS method was adopted and the determination was performed on Waters XTerra RP18（150 mm×4.6 mm,5 μm）column with acetonitrile-1 mmol·L-1 ammonium acetate buffer solution （pH adjusted to 9.0 using ammonia water）（35∶65） as mobile phase. Mass charge ratio of ion pair irbesartan was m/z -427.3→-193.0 and that of hydrochlorothiazide was m/z -296.0→-122.9. The bioequivalence of test preparation and reference preparation of Irbesartan hydrochlorothiazide tablets in 18 healthy volunteers after oral administration were investigated in randomized double-cycle cross-over design. RESULTS： The linear ranges were 2.00～5 000 μg·L-1 for irbesartan（r=0.998 4）and 2.00～200 μg·L-1 for hydrochlorothiazide （r=0.998 8）. The lowest limits of quantization were 2.00 μg ·L-1. The recovery rate was above 90%, and the intra-day and inter-day RSDs were both less than 10%. The recoveries of irbesartan ranged 96.3%～108.0%, and that of hydrochlorothiazide ranged 98.2%～100.5%. There were no significant difference in cmax, AUC0～t, AUC0～∞ between two preparation and cycles （P〉0.05）. There was no significant differenc in tmax between two preparations in non-parameter test （P〉0.05）. CONCLUSION： The method is simple in pretreatment and is specific, rapid, and accurate in analysis. The method can simultaneously determine plasma concentration of irbesartan and hydrochlorothiazide quantitatively. Two preparations are bioequivalent.