Flow injection determination of glucose, bile acid and ATP using immobilized enzyme reactor and chemiluminescent assay of NAD(P)H.
We have developed a chemiluminescent flow injection method for analysis of bile acid, glucose and ATP using the chemiluminescent assay of NADH using 1-methoxy-5-methylphenazinium methyl sulphate (1-MPMS)/isoluminol(IL)/microperoxidase (m-POD) system and immobilized enzyme reactors such as 3 alpha-hydroxysteroid dehydrogenase, glucose-dehydrogenase, hexokinase and glucose-6-phosphate dehydrogenase. The standard curves were obtained in the range of 5-100 pmol for bile acid, 0.5-5.0 nmol for glucose and 10(-7)-10(-5) mol/L for ATP. The coefficient of variation for each assay was not more than 4.1% for bile acid, 2.3% for glucose and 5.3% for ATP, respectively.