液相色谱-质谱联用法分析血液中氯胺酮和氟胺酮

Analysis of Blood-harbored Ketamine and F-Ketamine with LC-MS/MS

目的 通过考察基质效应筛选前处理方法,探索一种使用液相色谱 - 质谱联用法(LC-MS/MS)同时分析血液中氯胺酮和氟胺酮的方法。方法 取 1.00mL 血液,加入 2.00mL 乙腈,振荡提取,离心,取上清液过 0.22μm滤膜,上样分析。根据保留时间、特征离子对及其丰度比定性,根据特征离子峰面积定量。结果 乙腈沉淀蛋白法基质效应不显著,专属性好,血液中氯胺酮与氟胺酮浓度在 1.50 ~ 150ng/mL 范围内线性较好(R^(2)> 0.999),氯胺酮、氟胺酮方法检出限均为 0.30ng/mL,不同浓度添加样品回收率在 82.1% ~ 86.5% 之间,日内、日间RSD 均小于 10%。结论 本文成功建立了一种液相色谱 - 质谱联用法同时分析血液中氯胺酮与氟胺酮的方法,该方法操作简便、快捷、灵敏、稳定,适用于血液样品中氯胺酮、氟胺酮的定性定量检验。

Objective To establish a novel method for determination of blood-harbored ketamine and F-ketamine through liquid chromatography-mass spectrometry (LC-MS/MS), with pretreatment choice being preferentially determined from the matrix effect and successively the blood-harbored ketamine and F-ketamine to be isolated and tested of their specifi city,linearity, detection limit, precision and accuracy. Methods Sampling blood (1.00mL) was taken, added with 2.00mL acetonitrile, extracted under vibration, and centrifuged, having the supernatant filtered through 0.22μm membrane before analyzed into liquid chromatography-mass spectrometry (LC-MS/MS). With their retention time, characteristic ions and relative abundance ratio for qualitative determination plus their peak area of characteristic ions for quantifi cation, the two chemical substances were to characterize. Results Acetonitrile precipitation showed no significant matrix effect yet good specificity. A good linearity was obtained with the concentration of ketamine and F-ketamine ranging from 1.50ng/mL to150ng/mL in blood (R^(2)>0.999). The detection limit of the method was 0.30ng/mL, and the extraction effi ciencies of samples spiked with concentration-different ketamine or F-ketamine were 82.1%~86.5%. The intra- and inter-day RSDs were less than 10%. Conclusions A new method was here successfully developed for determination of blood-harbored ketamine and F-ketamine using liquid chromatography-mass spectrometry (LC-MS/MS). The method is simple, quick, sensitive and stable,suitable for qualitative and quantitative determination of ketamine and F-ketamine in blood.