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

发布时间：2018-07-03 15:17

本文选题：血小板活化因子 + 液质联用　；参考：《中国临床药理学杂志》2015年12期

【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1mmol·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·m L-1线性关系良好(r=0.998 4)。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。
[Abstract]:Objective to establish a LC-MS / MS method for the determination of platelet activating factor C-18 (PAF-C-18) and its acetylhydrolase (PAF-AH) activity in a novel drug allergic medium. Methods Platelet-activating factor C-18 was added to blank plasma and extracted with methanol, chloroform and ultrapure water. The chromatographic column was Agilent XDB C18, the organic phase was methanol containing 1mmol L-1 ammonium acetate, and the aqueous phase was methanol-acetonitrile-water 57: 20: 23 (containing 1 mmol L-1 ammonium acetate) for gradient elution. The specificity, standard curve and quantitative limit, precision and recovery, matrix effect and stability of the method were investigated. Results there was a good linear relationship between the peak of the sample at 19 min and the peak of the internal standard Lyso PAF C-16-d4 at 7 min. The peak of PAF C-18 was 0.5 ~ 25.0 ng mL ~ (-1) (r = 0.998 4). The RSD of intra-day and inter-day precision were below 10%. The recovery rate was 67.0% and 82.1%, the matrix effect was 115.3% and 116.9%, and the samples were completely metabolized at room temperature for 90 min, indicating that the samples should be treated immediately. Conclusion this method is a simple, rapid, specific and stable method for the determination of PAF-C-18 in human plasma, and can reflect the activity of PAF-AH indirectly by the decrease of PAF-C-18 in a certain time. It is convenient to carry out clinical pharmacological study of drug allergy.
【作者单位】：北京大学第三医院药剂科;北京大学治疗药物监测和临床毒理中心;北京大学临床研究所;
【分类号】：R927

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