1993 Volume 22 Issue 2 Pages 111-115
A fluorometric method for determining urinary estriol which is the principal estrogen in pregnancy has been developed. The method includes enzymatic hydrolysis of estrogen conjugates with β-glucuronidase from Ampullaria, extraction of free estrogen with dichloromethane followed by fluorometric analysis using a modified Kober color method.
Hydrolysis of estriol 16 β-glucuronide (E3 16G) was completed by an incubation with 1000 Fishman units of Ampullaria β-glucuronidase at 60°C for 5min. The calibration curve for E3 16G was linear in the wide range of 0.5-80mg/l. Reproducibility of within-run assay was CV 2.0-8.4% (n=10). Results obtained by this method were closely correlated with those by the other Kober reaction method and enzyme immunoassay. This method is applicable to the determination of estrogen in pregnancy urine, and useful for evaluation of feto-placental function.
