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Optimisation and Validation of a UHPLC-MS/MS Assay Using Timed Post-Column Ammonium Fluoride Infusion for Sensitive Serum Steroids Quantitation
bioRxiv · · Preprint · Open access
Dosedelova, Hawley + 4 more
Abstract ↗AI summary
The abstract is read at the publisher; the summary is JClub's.
A UHPLC-MS/MS serum assay quantifying 18 steroids achieved lower limits of quantitation from 0.04 to 2.1 nmol/L using timed post-column ammonium fluoride infusion.
- Why it matters: Limited ionisation efficiency and low circulating levels of some steroids constrain clinical steroid metabolomics, especially for low-abundance analytes.
- What they did: The method used supported liquid extraction from 100 L serum, then optimized ionisation by comparing formic acid versus post-column ammonium fluoride infusion, selecting 2 mmol/L timed infusion during separation.
- The result: Validation showed accuracy of 82–113% with intra- and inter-assay imprecision <10% and <13%, and healthy morning samples (55 women, 44 men) revealed sex-linked steroid concentration differences.
The findingWhy it mattersWhat they didThe result
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