A glutamate dehydrogenase-based method for the assay of L-glutamic acid: formation of pyridine nucleotide fluorescent derivatives.
A method for the quantitation of L-glutamic acid in the picomole range was developed by finding conditions which allowed the production of NADH by the action of the L-glutamate dehydrogenase (EC 1.4.1.3) and its subsequent transformation to a highly fluorescent derivative. The method measures linearly glutamate from 250 pmol to 5 nmol. For its simplicity and low cost it is ideally suited to the assay of a large number of samples within a single working day. Its application to the determination of regional glutamate levels in the rat brain, as well as to the measurement of ornithine aminotransferase (EC 2.6.1.13) activity from several tissues is described. The results are similar to those obtained by different methodologies in several laboratories, but the present method offers additional advantages.
Pérez-de la Mora
M
M
Departamento de Neurociencias, Universidad Nacional Autónoma de México, Distrito Federal.
Méndez-Franco
J
J
Salceda
R
R
Riesgo-Escovar
J R
JR
D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D005456:Fluorescent Dyes; D005969:Glutamate Dehydrogenase / Q000378:metabolism*; D005971:Glutamates / Q000032:analysis*; D018698:Glutamic Acid; D006834:Hydrazines; D006861:Hydrogen Peroxide; D008722:Methods; D009243:NAD / Q000378:metabolism; D009953:Ornithine-Oxo-Acid Transaminase / Q000032:analysis; D010084:Oxidation-Reduction; D051381:Rats; D012972:Sodium Hydroxide
D005456:Fluorescent Dyes; D005971:Glutamates; D006834:Hydrazines; D009243:NAD; C029424:hydrazine; D018698:Glutamic Acid; D012972:Sodium Hydroxide; D006861:Hydrogen Peroxide; D005969:Glutamate Dehydrogenase; C061421:glutamate dehydrogenase (NAD(P)+); D009953:Ornithine-Oxo-Acid Transaminase