Urinary Estrogens in Male Patients with Liver Diseases1. Studies on Method for the Estimation of Estrogens in Normal Male Subjects

In an attempt to establish optimal conditions for the hydrolysis of estrogen conjugates in urine with Brown's method, the study was made of the hot acid hydrolysis, enzyme (β-glucuronidase) hydrolysis and solvolysis. The modification of the micro-method according to Salokangas & Bulbrook was reinvestigated. urinary estrogens in fifteen normal male subjects were meseaured with some modified micro-method involving two step hydrolysis (β-glucuronidase hydrolysis and solvolysis). The following results were obtained. 1. Optimal conditions for the hot acid hydrolysis were in 15v/v % HCL concentration at 100℃ for 60minutes incubation. For β-glucuronidase hydrolysis, those were in 2,000units per ml enzyme concentration at 37℃ for 72hours incubation and at 50℃, incubation time was 48hours. Optimal incubation time for the solvolysis was 72hours at 37℃ in 2% H(2)SO(4) concentration with continuous ether extraction. 2. Compared with the hot acid hydrolysis and two step hydrolysis involving β-glucuronidase hydrolysis and solvolysis, quantities in total estrogens were mostly equal but estradiol fraction in the hot acid hydrolysis diminished little. 3. With the modified micro-method according to Salokangas & Bulbrook, the smallest amounts of pure estrogen could be estimated to 0.015μg and the mean ratios of recovery in this procedure were as follows; estrone 76.7%, estradiol 65.1%, estriol 82.8%. By this method, it was possible to dtermine urinary estrogens untill 0.5μg per day. 4. Urinary estrogens in eleven normal male subjects with 24 to 35years old were as follows; estrone 1.7±0.5μg per day (mean±standard deviation), estradiol 1.7±0.7, estriol 3.5±0.6, total estrogen 7.0±1.0, and in fourth with 60 to 70yers old, estrone 1.3±0.4μg per day, estradiol 2.5±0.4, estriol 2.6±1.1, total estrogen 6.4±1.2.