Determination of N-acetyl-L-glutamate using high-performance liquid chromatography.
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.
D000818:Animals; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) / Q000378:metabolism; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D004789:Enzyme Activation; D005969:Glutamate Dehydrogenase; D005971:Glutamates / Q000032:analysis*; D006868:Hydrolysis; D008099:Liver / Q000032:analysis; D008297:Male; D051379:Mice; D051381:Rats; D011919:Rats, Inbred Strains
D005971:Glutamates; D005969:Glutamate Dehydrogenase; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing); C016195:N-acetylglutamic acid