抄録
The procedure of gas-liquid chromatographic determination of urinary 11-deoxy-17-ketosteroids was divided into four steps in the present paper:
1) extraction and hydrolysis of 17-KS from urine in accordance with DePaoli's method, 2) purification of crude 11-deoxy-17-ketosteroids by thin-layer chromatography, 3) formation of trimethylsilyl ether derivatives, and 4) procedure of determination in gas-liquid chromatography.
A profitable and highly reproducible method for the clinical application was developed by modification of the single step hydrolysis method of glucuronides and sulfatesintroduced by DePaoli et al.
Recovery in extraction and hydrolysis steps using authentic steroids showed an average value of 90% for androsterone (Andro.), 89% for etiocholanolone (Etio.), and 91% for dehydroepiandrosterone (DHA).
Experimental error in gas-liquid chromatographic determination of trimethylsilyl ether derivatives remained within 1.6 % by six repeated determinations.
Study on reproducibility of the six repeated determinations, starting from the extraction step until the final chromatographic determination, was found to be satisfactory, by using pooled urine sample from normal male subjects. The coefficient variation showed 11% for Andro., 11% for Etio. and 14% for DHA.
The study on recovery for the whole procedure was carried out by adding a known amount of DHA sulfate to six samples of normal urine. The recovery was 67% in average, which agreed with the values in the literatures.
Thus, the method was applied to clinical analysis, and the values for adult and aged subjects were established. Studies on normal aged subjects disclosed significant depressions of Andro., DHA and total 11-deoxy-17-ketosteroids. Difference in Andro. accordingto sex which was observed in the normal adults was not in the aged subjects.
Urinary 11-deoxy-17-ketosteroids patterns were determined in patients with chronic hepatitis and with cirrhosis of the liver. There were reductions of Andro., Etio.and DHA under these pathological conditions, although no characteristic change in the steroid pattern was detected.
Determinations of the steroids in patients with hyperthyroidism were carried out. Increase in Andro./Etio. ratio was observed in this case. In addition, the results of triiodothyronine administration to the normal subjects revealed also enhancement of Andro./Etio. ratio, especially more remarkable in the aged subjects than in the adults. The above results suggest that triiodothyronine enhances 5α-reductase activity in the liver, and also suggest the decrease in enzymic activity of 5a-reductase in the aged subjects.
Steroid patterns were also determined in cancer patients. In patients with serum protein concentrations of less than 6 g/100 ml, total 11-deoxy-17-ketosteroids values were significantly lower than the values in patients with serum protein concentrations of 6g/100mland more. Any characteristic changes of steroid patterns, however, were not found out.
