Institute of Legal Medicine, Hannover Medical School (MHH), Hannover, Germany.
Anal Bioanal Chem. 2013 Aug;405(20):6595-7. doi: 10.1007/s00216-013-7074-z. Epub 2013 May 29.
To avoid the detection of small fragmentation products of γ-hydroxybutyrate (GHB), a liquid chromatography-tandem mass spectrometry GHB quantification method in human serum supported by adduct formation was developed and validated. The continuous infusion of GHB/GHB-D6 made the identification of two adducts possible and GHB/GHB-D6 sodium acetate adduct fragmentation was used as target mass transition. A Luna 5 μm C18 (2) 100 A, 150 mm × 2 mm analytical column and elution with a programmed flow of the mobile phase consisting of 10% A (H2O/methanol = 95/5, v/v) and 90% B (H2O/methanol = 3/97, v/v), both with 10 mM ammonium acetate and 0.1% acetic acid (pH = 3.2), were used. Protein precipitation with 1 mL of the mobile phase B was used as the sample preparation. The calculated limit of detection/quantification was 1 μg/mL. The presented study shows that the fragmentation of GHB sodium acetate adducts is an effective way of quantification of this small molecule and is an interesting alternative to other methods based on the detection of ions smaller than 85 Da. This fact together with the short analysis time of 3 min and the fast sample preparation make this method very attractive for forensic/clinical application.
为了避免 γ-羟基丁酸 (GHB) 的小碎片产物被检测到，开发并验证了一种支持加合物形成的人血清中 GHB 的液相色谱-串联质谱定量方法。GHB/GHB-D6 的连续注入使得两种加合物的鉴定成为可能，并且将 GHB/GHB-D6 醋酸钠加合物的片段化为目标质量过渡。使用 Luna 5 μm C18(2) 100 A，150 mm×2 mm 分析柱，洗脱程序为流动相组成的程序流量，流动相 A 为 10%（H2O/甲醇=95/5，v/v）和 90%B（H2O/甲醇=3/97，v/v），均含有 10 mM 醋酸铵和 0.1% 乙酸（pH=3.2）。使用 1 mL 流动相 B 进行蛋白质沉淀作为样品制备。计算的检测限/定量限为 1μg/mL。本研究表明，GHB 醋酸钠加合物的片段化是定量这种小分子的有效方法，并且是其他基于检测小于 85 Da 的离子的方法的有趣替代方法。这一事实加上 3 分钟的短分析时间和快速的样品制备，使该方法非常适合法医/临床应用。
