Development and validation of an LC-MS/MS assay for the quantification of allopregnanolone and its progesterone-derived isomers, precursors, and cortisol/cortisone in pregnancy

Development and validation of an LC-MS/MS assay for the quantification of allopregnanolone and its progesterone-derived isomers, precursors, and cortisol/cortisone in pregnancy

Neuroactive steroids are potent neuromodulators that play a critical role in both maternal and fetal health during pregnancy. These stress-responsive compounds are reportedly low in women with perinatal depression and may be associated with poor pregnancy outcomes in animal models. Chronic stress is...

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Bibliographic Details
Published in:Analytical and bioanalytical chemistry Vol. 413; no. 21; pp. 5427 - 5438
Main Authors: Mayne, G., De Bloois, E., Dabelea, D., Christians, U.
Format: Journal Article
Language:English
Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-09-2021
Springer
Springer Nature B.V
Subjects:
Analysis
Analytical Chemistry
Animal models
Assaying
Biochemistry
Characterization and Evaluation of Materials
Chemistry
Chemistry and Materials Science
Cortisol
Cortisone
Fetuses
Food Science
High performance liquid chromatography
Hormones
Hydrocortisone
Isomers
Laboratory Medicine
Liquid chromatography
LITAF protein
Mass spectrometry
Mass spectroscopy
Methods
Monitoring/Environmental Analysis
Neuromodulation
Neurosteroids
Postpartum depression
Pregnancy
Pregnanolone
Pregnenolone
Prenatal depression
Progesterone
Research Paper
Risk analysis
Risk factors
Steroid hormones
Steroids
United States
Pregnancy
Liquid chromatography–tandem mass spectrometry (LC-MS/MS)
Diagnostic
Biomarker
Neurosteroids
Allopregnanolone
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Summary:Neuroactive steroids are potent neuromodulators that play a critical role in both maternal and fetal health during pregnancy. These stress-responsive compounds are reportedly low in women with perinatal depression and may be associated with poor pregnancy outcomes in animal models. Chronic stress is a risk factor for adverse birth outcomes. Simultaneous quantification of neuroactive steroids, in combination with stress hormones cortisol/cortisone, provides an opportunity to investigate the synergistic relationship of these analytes within the convenience of one assay. A simple, reliable, and sensitive method for quantifying these endogenous compounds is necessary for further research with the potential to advance clinical diagnostic tools during pregnancy. Analytes were extracted from serum with a simple protein precipitation using methanol and then separated and quantified using high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS). After online extraction, analytes were separated using an Agilent Poroschell 120, 50 × 4.6 mm, 2.7 μm particle size, EC-C18 analytical column. The reliable quantification range was from 0.78 to 1000 ng/mL. QC sample inter- and intraday trueness was between 90 and 110% while inter- and intraday imprecision was less than 10%. Extracted samples were stable up to 7 days at 4 °C and extraction recovery was above 95%. Serum samples from 54 women in pregnancy were analyzed using this method. Here, we provide a validated, fast, and specific assay with sufficient sensitivity that allows for simultaneous quantification of blood serum concentrations of allopregnanolone (3α-hydroxy-5α-pregnan-20-one), pregnanolone (3α-hydroxy-5β-pregnan-20-one), epipregnanolone (3β-hydroxy-5β-pregnan-20-one), pregnenolone, progesterone, cortisol, and cortisone in pregnancy for clinical study samples and clinical diagnostics.
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ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-021-03523-0