Alonso E, Rubio V
Anal Biochem. 1985 Apr;146(1):252-9. doi: 10.1016/0003-2697(85)90423-3.
Acetylglutamate in HClO4 tissue extracts is first separated from glutamate by ion exchange. It is then deacylated with aminoacylase, and the resulting glutamate, after adsorption to and elution from an AG 50 column, is quantitated by a fast-HPLC method using o-phthaldialdehyde precolumn derivatization, separation in a C18 reverse-phase column, and fluorescence detection. A linear response is obtained up to 2 nmol, the detection limit is 5 pmol, and the method is suitable for assay in 1 mg liver tissue and thus for needle biopsies. When samples were analyzed by this procedure and by earlier procedures based upon detection of glutamate with glutamate dehydrogenase or upon activation of carbamoyl phosphate synthetase the results were similar. The method, which is highly specific, compares favorably in sensitivity, precision, and accuracy with all other published procedures. Using this assay, no acetylglutamate has been found in chicken liver and rat kidney.
高氯酸组织提取物中的乙酰谷氨酸首先通过离子交换与谷氨酸分离。然后用氨基酰化酶进行脱酰基反应，所得的谷氨酸在吸附到AG 50柱并从该柱洗脱后，通过一种快速高效液相色谱法进行定量，该方法采用邻苯二甲醛柱前衍生化、在C18反相柱中分离以及荧光检测。在2 nmol以内可获得线性响应，检测限为5 pmol，该方法适用于1 mg肝组织的测定，因此适用于针吸活检。当用该程序以及基于谷氨酸脱氢酶检测谷氨酸或基于氨基甲酰磷酸合成酶激活的早期程序分析样品时，结果相似。该方法具有高度特异性，在灵敏度、精密度和准确性方面与所有其他已发表的程序相比具有优势。使用该测定法，在鸡肝和大鼠肾中未发现乙酰谷氨酸。
