title: A novel spectrophotometric method for the enzymatic determination of NAD+ and NADH.
abstract: The theory and practice of a novel spectrophotometric method for the enzymatic determination of NAD+ and NADH is described. The method can not discriminate between NAD+ and NADH, but determines the concentration of the sum of both nucleotides. The method is based on the bleaching of p-nitroso-N,N-dimethylaniline (NMDA) (epsilon 440 nm = 35400 M-1cm-1) with NADH, in the presence of ethanol and yeast alcohol dehydrogenase, under the conditions of enzymatic cycling (ethanol > NDNA > NAD/H). The initial rates of -NDMA bleaching are proportional to the concentration of NAD+ or NADH, in a broad range from 10 nM to 100 microM.
authors.affiliation: Faculty of Technology, Bulevar Cara Lazara 1, Yugoslavia.
authors.forename: V
authors.initials: V
authors.lastname: Leskovac
authors.forename: S
authors.initials: S
authors.lastname: Trivić
authors.forename: B N
authors.initials: BN
authors.lastname: Anderson
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry; D006801:Humans; D007202:Indicators and Reagents; D007700:Kinetics; D009243:NAD / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D009603:Nitroso Compounds; D013056:Spectrophotometry, Ultraviolet
chemical_list: D007202:Indicators and Reagents; D009603:Nitroso Compounds; D009243:NAD; C008651:4-nitrosodimethylaniline; D000426:Alcohol Dehydrogenase