Schmid R, Karobath M
J Chromatogr. 1977 Sep 11;139(1):101-9. doi: 10.1016/s0021-9673(01)84130-3.
A gas chromatographic method for the determination of gamma-aminobutyric acid (GABA) in brain tissue is described. After microwave fixation, the brains were dissected and homogenized in 0.1 N formic acid; delta-amino-n-valeric acid (AVA), a homologue of GABA, was then added as an internal standard. After centrifugation, aliquots of the supernatant were treated with cation-exchange paper to adsorb the amino acids. The eluates of this paper were dried and the residues subjected to reaction with trifluoroacetic anhydride and hexafluoroisopropanol. After removal of the derivatization reagents by evaporation, the residues were dissolved in ethyl acetate and an aliquot was analysed by gas chromatography with electron-capture or mass fragmentographic detection. Quantitation can be carried out by either peak-height or peak-area measurements. The specificity of this method has been demonstrated with brain tissue by simultaneous mass fragmentographic analysis. The sensitivity is comparable to that of mass fragmentographic methods and is in the femtomole range. The method is simple and readily automated.
本文描述了一种用于测定脑组织中γ-氨基丁酸(GABA)的气相色谱法。经微波固定后,将大脑解剖并在0.1N甲酸中匀浆;然后加入GABA的同系物δ-氨基-n-戊酸(AVA)作为内标。离心后,取上清液的等分试样用阳离子交换纸处理以吸附氨基酸。将该纸的洗脱液干燥,残渣与三氟乙酸酐和六氟异丙醇反应。通过蒸发除去衍生化试剂后,将残渣溶解于乙酸乙酯中,取等分试样用带有电子捕获或质量碎片谱检测的气相色谱法进行分析。定量可通过峰高或峰面积测量进行。通过同时进行质量碎片谱分析已证明该方法对脑组织具有特异性。其灵敏度与质量碎片谱法相当,处于飞摩尔范围内。该方法简单且易于自动化。