Measurement of the ratio of primary to total bile acids in serum by enzymatic fluorometric microassay and its clinical significance in patients with liver disease

SHIRO IKAWA, HIRONAKA KAWASAKI, YASUHITO YAMANISHI, TETSUO MURA, MARIKO MIYAKE
1985 Tohoku journal of experimental medicine  
An enzymatic fluorometric microassay using 3a-or 7a-hydroxysteroid dehydrogenase was devised to determine serum total or primary bile acids. The present microassay using 50 ul of a serum sample was run as a two-step method ; the first step is the reaction of substrates with NAD+ and the hydroxysteroid dehydrogenase to produce as much NADH as possible in the incubation system and the second step is that of NADH with resazurin coupled with 1-methoxy phenazinemethosulfate to produce fluorescent
more » ... duce fluorescent resorfin. The assay is specific for 3a-or 7a-hydroxy bile acids, sensitive (a detection limit of 1.5 ,u M with a linear range of 1.5-80 p M), reproducible (CV = less than 5%) and inexpensive. As measured by this microassay, the ratio of serum primary to total bile acids (P/T ratio) in 10 normal subjects was 62.1+0.7% and was significantly elevated in the early phase of acute hepatitis (p <0.01), followed by return to the normal level after 4 weeks. enzymatic fluorometric microassay ; primary bile acids ; total bile acids ; liver disease
doi:10.1620/tjem.145.185 pmid:3857724 fatcat:dg53ftukqvd7xh7ei77txf642i