ABSTRACT
A systematic procedure for the extraction, separation, purification and colorimetric determination of urinary individual 17-ketosteroids in normal rhesus monkeys is described. Dehydroepiandrosterone was extracted after hot hydrolysis of the urine at neutral pH and the remaining steroid conjugates were extracted at pH 2 by the addition of ammonium sulphate. The liberated steroids were extracted after enzyme hydrolysis and the ketonic steroids were separated from the non-ketones by the Girard T reaction. The 11-deoxy steroids were separated from the 11-oxygenated steroids on a Celite column. They were further separated by paper chromatography and after elution, the individual 17-ketosteroids were estimated by the Zimmermann reaction. Normal mean excretion values in 30 female monkeys for total neutral 17-ketosteroids were 1.8 mg/24 h (range 1.5–2.6); total 17-hydroxycorticosteroids, 3.1 mg/24 h (range 2.2–4.8); androsterone, 38.6 μg/kg/24 h (range 9.3–60.2); aetiocholanolone, 19.2 μg/kg/24 h (range 6.8–30.7); dehydroepiandrosterone, 30.8 μg/kg/24 h (range 13.4–50.6); 11-ketoandrosterone, 7.5 μg/kg/24 h (range 5.1–14.1); 11-ketoaetiocholanolone, 10.2 μg/kg/24 h (range 6.8–18.3); 11β-hydroxyandrosterone, 15.4 μg/kg/24 h (range 8.8–26.7); 11β-hydroxyaetiocholanolone, 19.6 μg/kg/24 h (range 12.2–33.1). For total neutral 17-ketosteroids in 20 males, they were 2.5 mg/day (range 2.3–4.1); total 17-hydroxycorticosteroids, 3.7 mg/day (range 2.8–5.3); androsterone, 51.5 μg/kg/24 h (range 12.3–76.7); aetiocholanolone, 22.6 μg/kg/24 h (range 8.7–42.3); dehydroepiandrosterone, 41.3 μg/kg/24 h (range 16.9–59.7); 11-ketoandrosterone, 9.6 μg/kg/24 h (range 6.4–16.3); 11-ketoaetiocholanolone, 12.3 μg/kg/24 h (range 8.2–20.5); 11β-hydroxyandrosterone, 17.5 μg/kg/24 h (range 11.7–29.2); 11β-hydroxyaetiocholanolone, 21.7 μg/kg/24 h (range 14.5–36.4).
Studies on Milk Proteins. II. Colorimetric Determination of the Partial Hydrolysis of the Proteins in Milk
