Estimation of reference curves for the urinary steroid metabolome in the first year of life in healthy children: Tracing the complexity of human postnatal steroidogenesis

Abstract

Estimation of reference curves for the urinary steroid metabolome in the first year of life in healthy children: tracing the complexity of human postnatal steroidogenesis, Journal of Steroid Biochemistry and Molecular Biology http://dx.doi.org/10. 1016/j.jsbmb.2015.07.024 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.Page 1 of 24 Page 2 of 24 Graphical abstract HighlightsThe steroid metabolome during fetal-neonatal transition varies with sex and postnatal age.For 67 urinary steroid compounds normative values have been quantified by GC-MS for the first year of life.Highest concentrations are found for progesterone metabolites and glucocorticoids.In both sexes, most compounds peak at week 3 and decrease to 20% by week 25.Normative data for the urine steroid metabolome provide a powerful diagnostic tool. AbstractContext: Complex steroid disorders such as P450 oxidoreductase deficiency or apparent cortisone reductase deficiency may be recognized by steroid profiling using chromatographic mass spectrometric methods. These methods are highly specific and sensitive, and provide a complete spectrum of steroid metabolites in a single measurement of one sample which makes them superior to immunoassays. The steroid metabolome during the fetal-neonatal transition is characterized by a) the metabolites of the fetal-placental unit at birth, b) the fetal adrenal androgens until its involution 3-6 months postnatally, and c) the steroid metabolites produced by the developing endocrine organs. All these developmental events change the steroid metabolome in an age-and sex-dependent manner during the first year of life.Objective: The aim of this study was to provide normative values for the urinary steroid metabolome of healthy newborns at short time intervals in the first year of life. Methods:We conducted a prospective, longitudinal study to measure 67 urinary steroid metabolites in 21 male and 22 female term healthy newborn infants at 13 time-points from week 1 to week 49 of life. Urine samples were collected from newborn infants before discharge from hospital and from healthy infants at home. Steroid metabolites were measured by gas chromatography-mass spectrometry (GC-MS) and steroid concentrations corrected for urinary creatinine excretion were calculated.Results: 61 steroids showed age and 15 steroids sex specificity. Highest urinary steroid concentrations were found in both sexes for progesterone derivatives, in particular 20α-DH-5α-DH-progesterone, and for highly polar 6α-hydroxylated glucocorticoids. The steroids peaked at week 3 and decreased by ~80% at week 25 in both sexes. The decline of progestins, andro...

Current models of care for disorders of sex development – results from an International survey of specialist centres

“…Advances in biochemical analytical methods have led to an increase in the specificity and accuracy of measurement of steroid hormones and their metabolites in plasma and urine in the diagnostic work up of DSD [7–9]. Simultaneous developments in genetic and genomic technologies, combined with a marked reduction in costs, has provided a stimulus for change, with next-generation sequencing and whole-genome and -exome sequencing opening novel diagnostic strategies and expanding our knowledge of the underlying mechanisms of DSD [10].…”

How high-resolution techniques enable reliable steroid identification and quantification

“…We suggest that initial method development for challenging analytes as described in this paper should be performed on high-resolution instruments, then transferred to low-resolution instruments and re-validated for routine clinical applications. This recommendation results from own errors with low resolution analysis of baby urinary steroid metabolites 6 , detected by re-analysis with high-resolution techniques (see Tables 1 and 2).…”

“…5 Urine sample preparation was performed as originally described by Shackleton 1 as follows: (i) steroid extraction from urine, (ii) hydrolysis, (iii) second extraction of the steroids, (iv) derivatization and (v) sample clean-up prior to GC-MS and GCxGC-TOF MS analysis. 1,6 GC-MS analysis was performed by coupling a 7890A gas chromatograph to a 5795C mass spectrometer (both Agilent Technologies, Switzerland). Details of the GC-MS analysis have been previously published.…”

“…We therefore wanted to include in our high-resolution workflow routine panel the quantification of 22 additional progesterone metabolites in the urine of newborns as reported by Dhayat et al 6 For that, we re-analyzed 52 samples from the original study to evaluate the performance of GCxGC-TOF MS on these compounds, and to confirm correct analyte assignment. It is noteworthy that interpretation of the original GC-MS data was very cumbersome and some of the reported values were suspiciously high.…”

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“…The adrenal produces approximately two thirds of fetal cortisol towards the end of a pregnancy, with the remainder being derived maternally. Urinary steroids drop in normal neonates within the first week of life [21], after maternal-derived steroids have been excreted. If there was an inability of the neonatal adrenal gland to produce adequate cortisol then a compensatory rise in ACTH should be anticipated within the first week of life.…”