男子尿中 Estrogen の測定

R. I.・Ittrich-Fluorometry

抄録

Urinary estrogens have been assayed chiefly with the Kober hydroquinone colorimetry or sulfuric acid fluorimetry. However, it is difficult to determine accurately small amounts of estrogens such as in urines from males, oophorectomized women or menopausal women. In this study, male urinary estrogens were determined by a modification of the method reported by Kambegawa and some experimental studies were made on male urines. 200ml aliquot of 24-hour urine is hydrolyzed by the 3-step hydrolysis, added with estrone-6, 7³H, 17β-estradiol-6, 7³H and estriol-6, 7³H, and extracted with ether, followed by alkaline washes, methylation by a supersonic wave mixer, thin layer and column chromatography, liquid scintillation spectrometry and the Ittrich fluorometry. Results of experimental studies and estimation of urinary estrogens in males are as follows: 1) After the 3-step hydrolysis, 54.5% of the urinary estrogens were collected as the glucuronide, 30% in the solvolyzed fraction and 15.5% as the hot acid hydrolyzate. 2) The methylation of estrogens was achieved 10-20% better by the use of the supersonic wave mixer than by a magnetic stirrer. 3) Calculating efficiency of the Packard Tri-carb liquid scintillation spectrometer was tested in varieties of the estrogen concentrations using the dilution and the external standardization methods. There was noted no influence of the estrogen concentrations on the counting efficiency in the determination ranges. 4) In the range of 2×10^-3-1.0μg, a linear relationship on the standard curve was obtained applying the Allen's correction, when fluorometry was conditioned with the excitation light wave length of 510mμ and the fluorescence wave lengths of 530, 550 and 570mμ for each of the three estrogen-methyl ethers. 5) Urinary estrogens were determined in 15 normal males. In 5 subjects (2-13 years old) the values were: E₁: 1.06±1.09; E₂: 0.64±0.54; E₃: 0.76±0.68 and the total E: 2.46±1.99μg/24hr. In 5 men (22-38 years old), E₁: 5.02±1.57; E₂: 2.58±0.21; E₃: 4.11±1.82 and the total E: 11.72±2.97μg/24hr. In 5 men (46-72 years old), E₁: 3.81±0.31; E₁: 1.71±0.48; E₃: 3.51±0.94 and the total E: 9.03±1.33μg/24hr. 6) Urinary estrogens were also determined in patients with a variety of clinical syndromes.