Richard Damien, Ling Bing, Authier Nicolas, Faict Thierry W, Eschalier Alain, Coudoré François
Laboratoire de Toxicologie, Faculté de Pharmacie, Institut Régional de Médecine Légale, CHU G. Montpied, 63000 Clermont-Ferrand, France.
Anal Chem. 2005 Mar 1;77(5):1354-60. doi: 10.1021/ac048471h.
To quantify gamma-hydroxybutyrate (GHB) and its physiological metabolites, gamma-aminobutyric acid (GABA), 1,4-butanediol (1,4-BD), and gamma-butyrolactone (GBL) in various animal tissues (kidney, muscle, heart, liver, blood, brain cortex, thalamus, hypothalamus, hippocampus, or pons), an original gas chromatographic/mass spectrometric method with a automated solid-phase extraction by Oasis MCX cartridges on a Gilson Aspec Xli was developed. Using such apparatus allowed the limit of detection (LOD) of target compounds to be significantly lowered (LOD: 0.027, 0.025, and 5.7 microg/mL for GHB, 1,4-BD, and GABA, respectively, in 200 microL or microg of sample). After validation of each analytical step, the satisfactory performances of the apparatus in conjunction with the rapidity and ease of the extraction step make it suitable for simultaneous assay of GHB, 1,4-BD, GBL, and GABA. The method was used to test the correlation between GHB levels in tissues obtained at different times after death of male Sprague-Dawley rats and the postmortem interval. Preliminary results show a linear increase of GHB levels in relation to time of death in thoracic blood and central nervous system of animals kept at 15 and 20 degrees C.
为了定量分析各种动物组织(肾脏、肌肉、心脏、肝脏、血液、大脑皮层、丘脑、下丘脑、海马体或脑桥)中的γ-羟基丁酸(GHB)及其生理代谢物γ-氨基丁酸(GABA)、1,4-丁二醇(1,4-BD)和γ-丁内酯(GBL),开发了一种原始的气相色谱/质谱法,该方法采用吉尔森Aspec Xli上的Oasis MCX柱进行自动固相萃取。使用该仪器可显著降低目标化合物的检测限(对于200微升或微克样品中的GHB、1,4-BD和GABA,检测限分别为0.027、0.025和5.7微克/毫升)。在对每个分析步骤进行验证后,该仪器令人满意的性能以及萃取步骤的快速性和简便性使其适用于同时检测GHB、1,4-BD、GBL和GABA。该方法用于测试雄性斯普拉格-道利大鼠死亡后不同时间获得的组织中GHB水平与死后间隔之间的相关性。初步结果表明,在15和20摄氏度下保存的动物的胸腔血液和中枢神经系统中,GHB水平随死亡时间呈线性增加。