高效液相质谱联用法测定人血浆中蒿甲醚及其活性代谢物双氢青蒿素的浓度(英文)

Determination of artemether and its metabolite dihydroartemisinin in human plasma by HPLC with atmospheric pressure chemical ionization mass spectrometry

目的建立高效液相质谱联用法以测定人血浆中蒿甲醚(ARM)及其活性代谢物双氢青蒿素(DHA)的浓度.方法色谱条件:色谱柱为C18(150 mm×4.6 mm,5μm),流动相为0.2%醋酸-甲醇梯度洗脱系统,流速为1.0 mL·min-4.质谱条件:大气压化学电离方法(APCI)采集正离子,SIM方式对ARM和DHA检测m/z 221,对内标(青蒿素)检测m/z 283.血浆样品的预处理采用甲基叔丁基醚萃取法.结果ARM和DHA的线性范围为5～300μg·L-1,相关系数r＞0.999 0,检测限均为2μg·L-11.ARM和DHA日内和日间测定的RSD＜9.3%,回收率在92%～105%的范围内,提取回收率为80%～96%.结论该法简便、准确、灵敏、专属,适用于ARM和DHA的人体药动学研究.

AIM To develop a HPLC-MS method for the determination of ARM and its active metabolite DHA in human plasma. METHODS The chromatography separation was performed on a C18 column （150 mm × 4.6 mm, 5 μm） with a gradient elution mobile phase consisting of 0.2 % glacial acetic acid aqueous solution and methanol at a flow rate of 1.0 mL·min^-1 .MS detection in SIM mode using APCI as an interface was applied to determine positive ions at m/z 221 for ARM and DHA, and m/z 283 for internal standard （artemisinin）. Plasma samples were extracted with methyl t- butyl ether, followed by evaporation of the supernatant to dryness and reconstitution of the residue in methanol. RESULTS High specificity and a low detection limit （2 μg· L^-1） were achieved for ARM and DHA. Linearity was confirmed in the concentration range of 5 -300 μg· L^-1 with the correlation coefficient （r） 〉 0.999 0. The RSD values of intra - and inter-day assay for both compounds were less than 9.3 % and the recoveries were within 92 % - 105 % . The extraction recovery of ARM and DHA ranged from 80% to 96%. CONCLUSION The tive and can be applicable to pharmacokinetic studies of ARM and DHA in method proves to be simple, accurate, sensi- humans.