Steroids in Meconium from Male and Female Newborn Infants

Abstract

The steroids in meconium from normal male and female newborn infants have been studied by gas chromatography-mass spectrometry. Practically all steroids were excreted as monoand disulphates. The approximate concentrations of steroid mono-and disulphates were 430 and 470pg per g of meconium. Twenty unsaturated and fourteen saturated steroids were completely identified. Most of the unsaturated steroids had a 3/?-hydroxy-A6 structure. I n addition, seven saturated steroids were tentatively identified. No significant qualitative differences were found in the steroid composition of meconium from male and female newborns. The results are discussed in relation to present knowledge of steroid metabolism in the foeto-placental unit.Very little information is available on the excretion of neutral steroid metabolites in meconium from newborn infants. I n a series of publications Francis et al. . In order to obtain a complete understanding of the metabolism of neutral steroid hormones in the foeto-placental unit it was considered of interest to have a detailed knowledge of the metabolites excreted in meconium. New methods of purification in combination with gas chromatographymass spectrometry (for references see [6]) have now made it possible to separate and identify even minor constituents in small amounts of biological material. MATERIALS AND METHODS Collection of MeeoniumMeconium was collected from diapers not contaminated with urine. Samples from 6 male and 9 female infants were pooled to give a meconium pool from males (70 g) and a meconium pool from females (40 g). The samples were stored a t -20 "C until analysed. SteroidsThe sources of several of the steroids used as reference compounds were given in a previous Unusual Abbreviations. Progesterone, silyl, trimethylsilyl. The terms pregnanolone and pregnanediol are used as collective names for isomers of 3-hydroxy17p-pregnan-20-one and pregnane-3,2O-diol, respectively.Enzymes. 3-a-Hydroxysteroid : NADP oxidoreductase (EC 1.1.1.50).Note. Retention times, tg, are calculated relative to 5a-cholestane.paper [7]. 3~-Hydroxy-5-androsten-l7-one was purchased from Ikapharm (Ramat-Gan, Israel). 3/?,78-Dihydroxy-5-androsten-17-one and 3B,18-dihydroxy-5-androsten-17-one were gifts from Dr. G. Johannsson and Dr. C. H. L. Shackleton, respectively. 5-Androstene-3/?,17a-diol and 5m-androstane-3a,17a-diol were obtained from Schering AG (Berlin, Germany). 38-2 1 -Dihydroxy-5-pregnen-20 one-21 -acetate (purchased from Sigma Chemical Company, St. Louis, Missouri, U.S.A.) was hydrolysed with K2C0, in methanol a t room temperature over night. The resulting 38,21 -dihydroxy-5-pregnen-ZO-one was reduced with sodium borohydride in methanol and the following compounds were found : 5-pregnene-3@,20B, 21 -trio1 (high yield) and 5-pregnene-3B,20a,21 -trio1 (low yield). 3a-Hydroxy-5a-pregnan-ZO-one, 3m-hydroxy-5B-pregnan-ZO-one and 3B-hydroxy-5-pregnen-20-one were purchased from Ikapharm (Ramat-Gan, Israel). Sodium borohydride reductions of these compounds gave the corresponding 208-and 20a-hydr...

Identification and Measurement of Sulphate-Conjugated Neutral Steroids in the Intestinal Contents of Early and Mid-Term Foetuses

“…The TMS ether of this compound had a mass spectrum identical with that of a 3g,15£,16£-trihydroxy-5g-pregnan-20-one found in newborn meconium by Gustafsson & Stenberg (1971). The mass spectra of the TMS ethers of compound 27 and 3/?,15a,16a-trihydroxy-5a-pregnan-20-one were very similar (Eriksson & Gustafsson, 1970).…”

“…The first neutral steroids identified in this source were dehydroepiandrosterone (Francis, Shen & Kinsella, 1960), 3a-hydroxy-5/?-pregn-16-en-20-one (Francis, Shen & Kinsella, 1962), 5/?-pregnane-3a,20a-diol (Francis & Kinsella, 1966) and 5a-pregnane-3a,20a-diol (Francis & Kinsella, 1967). Quite recently, analyses by gas chromatography and gas chromatography-mass spectro¬ metry have clearly shown the complexity of the steroid pattern in the meconium Gustafsson & Sjövall, 1970;Gustafsson & Stenberg, 1971; Kinsella & Francis, 1971). …”

Origin of 5α‐pregnane‐3α,20α,21‐triol 3‐sulphate in pregnant women

“…Sulphates of pregnane-3,20,21-triols have only been found in biological fluids from pregnant women [6] and from newborn infants [6,25]. Our findings indicate that 21-hydroxylation of sulphated C2102 steroids is an important metabolic pathway in pregnancy.…”

Studies on Steroids in Urine of the Male Newborn

“…It had been isolated before from equine urine (59). Recently, it was again described in meconium and feces (34,60,74,75) and isolated from human fetal bile (76). Trace amounts have been described in newborn urine (9) and it seems to be utilized as precursor in steroid biosynthesis (61).…”