高效液相色谱串联质谱法测定人血浆中美金刚浓度

Development of memantine in human plasma by liquid chromatography tandem mass spectrometry

目的:建立测定人血浆中美金刚浓度的高效液相色谱串联质谱法,以用于美金刚人体药动学研究。方法:Ultimate XB-C18柱(150mm×4.6mm,5μm)为色谱柱,甲醇-0.5%甲酸(70∶30)为流动相,流速为0.5mL·min-1;电喷雾电离正离子模式,选择性监测质荷比(M/Z)为180→163(美金刚)和152→135(金刚烷胺,内标);血浆样本经醋酸乙酯提取后进行分析。结果:血浆中美金刚质量浓度在0.2～100μg·L-1范围内线性关系良好,最低定量限达0.2μg·L-1;日内、日间精密度RSD小于7,血浆样本经冷冻和冻融后均稳定。结论:此方法灵敏、专一、快速、简便,适用于盐酸美金刚胶囊人体药动学研究。

OBJECTIVE To validate a high-performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method for the determination of memantine in human plasma. METHODS The analyte was separated on a Ultimate XB-C18 column（150 mm 4. 6 mm, 5 μm） with a mobile phase of methanol and water containing 0. 5% formic acid （70：30） at a flow rate of 0. 5mL·min^-1. The multiple reaction monitoring （MRM） mode was used for analyte quantification, and the precursor-to-prod- uct ion transition of m/z 180→163 （memantine） and 152→135 （amantadine, IS） were selected, respectively. The plasma sample was liquid-liquid extracted by ethyl acetate. RESULTS The weighted （ 1/x^2） calibration curve was linear in the 0.2 - 100. 0 μg·L^-1 range for memantine, and the low limit of quantification was 0. 2 μg·L^-1. The inter- and intra-day precisions （RSD） were 〈7. Plasma samples were stable under freezed and freeze-thaw conditions. CONCLUSION The developed LC/ MS/MS method was selective, sensitive and simple, and was successfully applied to the pharmacokinetic study of memantine after oral administration to Chinese healthy volunteers.