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

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Alternate steroid sulfation pathways targeted by LC-MS/MS 1 analysis of disulfates. Application to prenatal diagnosis of steroid synthesis disorders. / Pozo, Oscar J.; Marcos, Josep ; Khymenets, Olha ; Pranata, Andy; Fitzgerald, Christopher C. ; McLeod, Malcolm D. ; Shackleton, Cedric.

In: Journal of Molecular Endocrinology, 19.02.2018.

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@article{d29b220c76ca46c2a276a4ec7d95a283,
title = "Alternate steroid sulfation pathways targeted by LC-MS/MS 1 analysis of disulfates. Application to prenatal diagnosis of steroid synthesis disorders",
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.",
keywords = "Steroid bis-sulfates, steroid disulfates, steroid sulfation, prenatal diagnosis, LC-MS/MS, steroid metabolomics",
author = "Pozo, {Oscar J.} and Josep Marcos and Olha Khymenets and Andy Pranata and Fitzgerald, {Christopher C.} and McLeod, {Malcolm D.} and Cedric Shackleton",
year = "2018",
month = feb,
day = "19",
doi = "10.1530/JME-17-0286",
language = "English",
journal = "Journal of Molecular Endocrinology",
issn = "0952-5041",
publisher = "BioScientifica",

}

RIS

TY - JOUR

T1 - Alternate steroid sulfation pathways targeted by LC-MS/MS 1 analysis of disulfates. Application to prenatal diagnosis of steroid synthesis disorders

AU - Pozo, Oscar J.

AU - Marcos, Josep

AU - Khymenets, Olha

AU - Pranata, Andy

AU - Fitzgerald, Christopher C.

AU - McLeod, Malcolm D.

AU - Shackleton, Cedric

PY - 2018/2/19

Y1 - 2018/2/19

N2 - 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.

AB - 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.

KW - Steroid bis-sulfates

KW - steroid disulfates

KW - steroid sulfation

KW - prenatal diagnosis

KW - LC-MS/MS

KW - steroid metabolomics

U2 - 10.1530/JME-17-0286

DO - 10.1530/JME-17-0286

M3 - Article

JO - Journal of Molecular Endocrinology

JF - Journal of Molecular Endocrinology

SN - 0952-5041

ER -