Determination of pregnane-17,21-diol-20-ones in human urine

Treatment of urine or of a urine extract with periodate followed by treatment of the acidic oxidation products with sodium bismuthate gave a mixture of neutral end products consisting only or predominantly of 17-oxoandrostanes derived from the corresponding urinary pregnane-17,21-diol-20-ones. The mixture of end products was resolved by gas-liquid or paper chromatography. The latter technique was used to determine cortisol secretion rates and the results were found to be in reasonable agreement with those obtained by a double-isotope method. Gas-liquid-chromatographic characterization of steroidal 4-en-3-ones is described.     

A simple radioimmunoassay for plasma cortisol and 11-deoxycortisol (17, 21-dihydroxy-4-pregnene-3, 20-dione).

A simple procedure for the measurement of cortisol and 11-deoxycortisol in plasma or serum is described. One-half ml. of plasma is extracted with methylene chloride. Separation of cortisol and deoxycortisol is achieved by a Sephadex LH-20 mini-column. The assay itself is achieved by the use of a commercially available cortisol kit employing rabbit anti-cortisol coated tubes. This antibody exhibits a 20% crossreactivity with 11-deoxycortisol. This procedure is extremely useful in the assessment of a pituitary-adrenal reserve in the Metyrapone test.