血小板活化因子C-18及其乙酰水解酶的液质联用检测方法  

LC-MS/MS method for the detection of platelet-activating factor C-18 and its acetylhydrolase

作　　者：刘维[1,2] 熊歆[1,2] 宋福鱼[3] 杨丽[1,2] 翟所迪[1,2] 

机构地区：[1]北京大学第三医院药剂科,北京100191 [2]北京大学治疗药物监测和临床毒理中心,北京100191 [3]北京大学临床研究所,北京100191

出　　处：《中国临床药理学杂志》2015年第12期1191-1193,1201,共4页The Chinese Journal of Clinical Pharmacology

摘　　要：目的：建立LC－MS／MS法检测新型药物过敏反应介质血小板活化因子C－18（PAF C－18）及其乙酰水解酶（PAF－AH）活性的方法。方法在空白血浆中添加血小板活化因子C－18标准品，用甲醇、氯仿及超纯水进行提取，取出下层，氮气吹干复溶后进样。色谱柱为 Agilent XDB C18，有机相为含有1 mmol· L－1醋酸铵的甲醇，水相为甲醇－乙腈－水＝57∶20∶23（含有1 mmol· L－1醋酸铵），进行梯度洗脱。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、基质效应及稳定性。结果样品在19 min出峰，内标Lyso PAF C－16－d4在7 min出峰。 PAF C－18在0．5～25．0 ng· mL－1线性关系良好（ r ＝0．9984）。日内及日间精密度 RSD 均在10％以下。回收率在67．0％～82．1％，基质效应在115．3％～116．9％，于室温放置90 min即完全代谢，说明样本需立即处理。结论本方法是一种简单、快速、特异、稳定地测定人血浆中PAF C－18的方法，并可以通过外源性添加PAF C－18在一定时间内的下降程度，间接反映PAF－AH活性，便于开展药物过敏的临床药学研究。Objective To establish a liquid chromatography -mass spectrometry （ LC-MS/MS） method for the analysis of platelet-activa-ting factor C-18（PAF C-18）and its acetylhydrolase in human plasma. Methods A convenient extraction procedure using trichloromethane, methanol and water as the extract solvent was used.The separation was carried out on Agilent XDB C -18 analytical column.Mobile phase A consisted of methanol -ammonium acetate（1 mmol· L-1 ）-acetonitrile （ 57 ∶23 ∶20 ） , and mobile phase B was methanol containing 1 mmol· L-1 ammonium acetate.Gradient elution was used.The speci-ficity, linearity and limit of quantification, precision and recovery, ma-trix effect and stability were assessed.Results The assay was reproduci-ble within the linearity range from 0.5-25.0 ng· mL-1 （ r=0.998 4 ） . The relative standard deviation（ RSD） of intra-day precision were below 10%, with recovery rate ranged from 67.0% to 82.1% and the matrix effect ranged from 115.3%to 116.9%.PAF C-18 was fully degraded over 90 min at room temperature.Conclusion This method provides a sensitive, special and convenient approach for the clinical analysis of PAF C-18, as well as the indirect measurement of PAF-acetylhydrolase activity by analyzing the concentration decrease of an exogenous PAF.

关 键 词：血小板活化因子 液质联用 血小板活化因子乙酰水解酶 

分 类 号：R969.3[医药卫生—药理学]