Alternate steroid sulfation pathways targeted by LC-MS/MS 1 analysis of disulfates. Application to prenatal diagnosis of steroid synthesis disorders

Research output: Contribution to journalArticle

Authors

  • Oscar J. Pozo
  • Josep Marcos
  • Olha Khymenets
  • Andy Pranata
  • Christopher C. Fitzgerald
  • Malcolm D. McLeod

Colleges, School and Institutes

External organisations

  • Integrative Pharmacology and Systems Neuroscience Group, IMIM, Hospital del Mar, Doctor Aiguader 88, Barcelona, Spain
  • Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Doctor Aiguader 88, 08003 Barcelona, Spain.
  • Cerba Internacional, Pl. Ramon Llull, 7, 08203 Sabadell, Spain.
  • Research School of Chemistry, Australian National University, Canberra, ACT 2601, Australia

Abstract

The steroid disulfates (aka bis-sulfates or bis(sulfates)) are a significant but minor fraction of the urinary steroid metabolome that have not been widely studied because major components are not hydrolyzed by the commercial sulfatases commonly used in steroid metabolomics. In early studies, conjugate fractionation followed by hydrolysis using acidified solvent (solvolysis) was used for the indirect detection of this fraction by GC-MS. This paper describes the application of a specific LC-MS/MS method for the direct identification of disulfates in urine, and their use as markers for the prenatal diagnosis of disorders causing reduced estriol production: STSD (Steroid Sulfatase Deficiency), SLOS (Smith-Lemli-Opitz Syndrome) and PORD (P450 Oxido-Reductase Deficiency). Disulfates were detected by monitoring a constant-ion-loss (CIL) from the molecular di-anion. While focused on disulfates, our methodology included an analysis of intact steroid glucuronides and monosulfates because steroidogenic disorder diagnosis usually requires an examination of the complete steroid profile. In the disorders studied, a few individual steroids (as disulfates) were found particularly informative: pregn-5-ene-3β,20S-diol, pregn-5-ene-3β,21-diol (STSD, neonatal PORD) and 5α-pregnane-3β,20S-diol (pregnancy PORD). Authentic steroid disulfates were synthesized for use in this study as aid to characterization. Tentative identification of 5ξ-pregn-7-ene-3ξ,20S-diol and 5ξ-pregn-7-ene-3ξ,17,20S-triol disulfates was also obtained in samples from SLOS affected pregnancies. Seven ratios between the detected metabolites were applied to distinguish the three selected disorders from control samples. Our results show the potential of the direct detection of steroid conjugates in the diagnosis of pathologies related with steroid biosynthesis.

Details

Original languageEnglish
Number of pages38
JournalJournal of Molecular Endocrinology
Early online date19 Feb 2018
Publication statusE-pub ahead of print - 19 Feb 2018

Keywords

  • Steroid bis-sulfates, steroid disulfates, steroid sulfation, prenatal diagnosis, LC-MS/MS, steroid metabolomics