The partial characterization of 1-oxygenated steroids from urine of a hypertensive newborn child

Abstract

1. Four substances from the urine of a hypertensive newborn girl were partially characterized and shown to be 17oc-hydroxy-5, B-pregnane-1,3,20-trione, 3oc,17a-dihydroxy-5,B-pregnane-1,20-dione, 3a,17oc,20ac-trihydroxy-5p-pregnan-1-one and 5/3-pregnane-lB,3oc,17a,20c-tetrol. 2. The characterization rested mainly on RM analysis of the substances and their derivatives by glycol fission, providing evidence for position and degree of substitution and for steroidal character. Supporting evidence was provided by chemically specific location reactions. 3. Certain problems in the manipulation of these ,B-disubstituted steroids are discussed.The high 17-hydroxy corticosteroid excretion of patient S.C. when 1-3 weeks old and grossly hypertensive has been described elsewhere (Edwards, Harvey & Knight-Jones, 1968 MATERIALS AND METHODSFull details of the procedures used for the isolation of urinary steroids are described elsewhere (Edwards, 1968a) and in the present instance consisted of the following steps. Urine samples (6 days' collection, age between 8 and 20 days) from S.C. during her period of high 11-oxygenation index (ratio of metabolites of 17-hydroxylated precursors of cortisol to metabolites of cortisol; Edwards, Makin & Barratt, 1964) (Stevens, 1964). Each substance was homogeneous on the criterion that portions from the forward and rear portions ofa chromatographed zone on rechromatography gave zones side by side of identical RF values.The chemical transformations used were (i) borohydride reduction and (ii) periodate oxidation, both carried out as described by Edwards et al. (1964), and (iii) oxidation with Jones reagent carried out as described by Djerassi, Engle & Bowers (1956), but with trebled proportion of acetone. RM analysis procedure. RM values were calculated from RF values and referred to the root system of the family (light petroleum-methanol-water, 5:4:1, by vol.) as RMO values by correcting for the effects of benzene replacing light petroleum (Kabasakalian & Basch, 1960;Edwards, 1968a). Observed ARM values were compared with reference values (Table 1) and the latter used to indicate likely structural changes. For clarity the ARM value observed in changing from substance of structure A to structure B was signified ARM [A--B]. It was presumed that substances with RMO values differing by less than 0-07 (S.D. ofrepetition of a value on different occasions) were in fact one substance.Referencel-hydroxysteroid. loc-Hydroxy-5oc-androstane-3,17-dione samples from Dr Fukushima and Dr Foell were found to be identical with the exception of a small proportion of contaminant in the latter. This was assumed to be the lfl-isomer, since other isomerization was unlikely from the method of preparation. A similar contaminant was present in the lac-hydroxyandrost-4-ene-3,17-dione sample. The lc,3,B-dihydroxy-50c-androstan-17-one sample was uncontaminated.Other reference steroids were prepared by unambiguous chemical transformations and were purified by chromatography, but there was insufficient material available...

Congenital Adrenal Hyperplasia and Other Conditions Associated With a Raised Urinary Steroid 11-Oxygenation Index

“…Their main presenting features were virilization and hyper¬ tension without salt loss, all of which regressed spontaneously (Edwards, Harvey & Knight-Jones, 1968) or after surgical removal of an adrenal mass (Edwards, Snodgrass & Daly, 1971). This interference can be eliminated by studies of the 1,3,17,20-tetraoxygenated pregnane derivatives themselves (Edwards & Trafford, 1968) or by determination of the ll-OI value by GLC (Makin, 1970). The reason for inter¬ ference by 1-oxygenated steroids is that the product in the borohydride reduction, periodate oxidation and esterification is a lg,3a-diformyloxy-17-oxosteroid which appears on chromatograms with aetiocholanolone formate.…”