Nahar Limon Khatun, Cordero Rosa Elena, Nutt David, Lingford-Hughes Anne, Turton Samuel, Durant Claire, Wilson Sue, Paterson Sue
Toxicology Unit, Imperial College London, London W6 8RP, UK
Toxicology Unit, Imperial College London, London W6 8RP, UK.
J Anal Toxicol. 2016 Mar;40(2):117-23. doi: 10.1093/jat/bkv125. Epub 2015 Nov 4.
A highly sensitive and fully validated method was developed for the quantification of baclofen in human plasma. After adjusting the pH of the plasma samples using a phosphate buffer solution (pH 4), baclofen was purified using mixed mode (C8/cation exchange) solid-phase extraction (SPE) cartridges. Endogenous water-soluble compounds and lipids were removed from the cartridges before the samples were eluted and concentrated. The samples were analyzed using triple-quadrupole liquid chromatography-tandem mass spectrometry (LC-MS-MS) with triggered dynamic multiple reaction monitoring mode for simultaneous quantification and confirmation. The assay was linear from 25 to 1,000 ng/mL (r(2) > 0.999; n = 6). Intraday (n = 6) and interday (n = 15) imprecisions (% relative standard deviation) were <5%, and the average recovery was 30%. The limit of detection of the method was 5 ng/mL, and the limit of quantification was 25 ng/mL. Plasma samples from healthy male volunteers (n = 9, median age: 22) given two single oral doses of baclofen (10 and 60 mg) on nonconsecutive days were analyzed to demonstrate method applicability.
开发了一种高灵敏度且经过充分验证的方法，用于定量测定人血浆中的巴氯芬。使用磷酸盐缓冲溶液（pH 4）调节血浆样品的pH后，采用混合模式（C8/阳离子交换）固相萃取（SPE）柱对巴氯芬进行纯化。在洗脱和浓缩样品之前，从柱上去除内源性水溶性化合物和脂质。使用三重四极杆液相色谱-串联质谱（LC-MS-MS），采用触发式动态多反应监测模式进行分析，以同时进行定量和确证。该测定法在25至1000 ng/mL范围内呈线性（r(2) > 0.999；n = 6）。日内（n = 6）和日间（n = 15）精密度（相对标准偏差百分比）<5%，平均回收率为30%。该方法的检测限为5 ng/mL，定量限为25 ng/mL。分析了9名健康男性志愿者（中位年龄：22岁）在非连续日期接受两次单剂量口服巴氯芬（10和60 mg）后的血浆样品，以证明该方法的适用性。
