Abstract
A highly sensitive chemiluminescence assay for NAD(P)H have been developed. The principle of present method is as follows; NAD(P)H reduced molecular oxygen to O2- and hydrogen peroxide (H2O2) in the presence of 1-methoxy-5-methylphenazinium methylsulfate (1-MPMS) as electron mediator. The produced O2- and H2O2 could be assayed by chemiluminescence reaction using isoluminol (IL)-microperoxidase (m-POD). A linear relationship between chemiluminescence intensity and NAD(P)H (log/log) was obtained ranged from 10-9 M to 10-5 M, and the detection limit was about 100 fmol/assay. The relative standard deviation of NADH and NADPH were 1.15.8 and 1.47.6%, respectively. This chemiluminescence reaction has been applied to the determination of total bile acids in serum, enzyme activity of glucose-6-phosphate dehydrogenase and glucose in blood. Plasma samples were assayed for total bile acids and glucose by both conventional and present method. The results showed good agreement by the correlation coefficient {r=0.96 for total bile acids (n=83) and r=0.99 (n=100) for glucose}.
