LC-MS/MS法测定人血浆中的金刚烷胺浓度及其药动学

Determination of amantadine in human plasma by LC-MS/MS and its pharmacokinetics

目的建立HPLC—MS／MS分析方法测定人血浆中的金刚烷胺浓度，并用于研究健康受试者口服氨酚咖黄烷胺片后金刚烷胺的药动学。方法血浆经乙醚一二氯甲烷（4：1，V／y）提取，采用C18柱为分析柱，甲醇-10mmol·L^-1醋酸铵溶液一冰醋酸（60：40：0．08，V／V／V）为流动相，流速1．0mL·min^-1，柱后分流，0．2mL·min^-1进入质谱，柱温35℃。质谱检测方式：多反应离子监测，选择监测的离子为m／z 152→m／z 135（金刚烷胺）和m／z275→m／z 230（内标物，氯苯那敏）。结果金刚烷胺的线性范围为1—200μg·L^-1（r=0．9999），血浆中金刚烷胺的最低定量限达1μg·L^-1（RS,N〉10），回收率〉80％。结论本法简便、准确、灵敏度高，适用于金刚烷胺的药动学研究。

AIM To develop an LC-MS/MS assay to determine amantadine in human plasma and investigate the pharmacokinetics of amantadine after oral administration of the compound preparation. METHODS The plasma samples were extracted with ethyl ether-dichlormethane （4： 1, V/V）. The separation was carried out on a Shimadzu ODS column （150 mm × 4.6 mm, 5μm） at 35 ℃. The mobile phase consisted of methanol, 10 mmol·L^-1 ammonium acetate and acetic acid （60：40：0.08, V/V/V）, runrfing at a flow rate of 1.0 mL·min^-1. After separation, the mobile phase was splitted and 0.2 mL·min^-1 was into MS. LC-MS/MS was performed in the MRM mode using target ions at m/z 152→ m/z 135 for amantadine and m/z 275→m/z 230 for chlorphenamine. RESULTS The linear calibration curve was obtained in the concentration range of 1 -200μg· L^-1（ r = 0.999 9）. The lowest limit of quantification of amantadine in plasma was 1μg· L^-1. The average recovery was more than 80%. CONCLUSION The assay is proved to be sensitive, accurate and convenient. The method can be used for the pharmacokinetic research of amantadine.