{
  "abstract": "Steroid metabolomics has potential for identifying biomarkers and improving disease diagnosis. However, poor ionisation efficiency and low circulating concentrations of some steroids limit the broader clinical applications. Here, we developed a ultra-high-performance liquid chromatography - tandem mass spectrometry (UHPLC-MS/MS) assay for quantitation of 18 steroids in serum encompassing the mineralocorticoid, glucocorticoid and androgen biosynthesis pathways, including both classic and 11-oxygenated androgens. Steroid extraction from 100 L of serum was performed using supported liquid extraction with ethyl acetate elution. Ionisation efficiency of steroids was investigated using formic acid in the mobile phase and post-column infusion of ammonium fluoride. Ammonium fluoride enhanced the signal intensity of 3-keto-{Delta}4 steroids up to ninefold, however A-ring reduced steroids exhibited greater ionisation with formic acid. Thus, to optimise sensitivity, a timed post-column infusion of 2 mmol/L ammonium fluoride was employed during chromatographic separation. The lower limits of quantitation ranged from 0.04 to 2.1 nmol/L. Accuracy ranged from 82 to 113% and intra- and inter-assay imprecision was <10% and <13%, respectively. The assay was applied to analyse morning serum steroid concentrations in healthy individuals (55 women, 44 men; age-matched). Serum progesterone, 11-deoxycorticosterone, dehydroepiandrosterone and androstenedione were significantly higher in women; in contrast, testosterone and 5-dihydrotestosterone were higher in men. In summary, we have successfully developed and validated a UHPLC-MS/MS assay for multi-steroid profiling ad applied it to healthy individuals of both sexes.",
  "abstract_shown": true,
  "affiliations": [
    "MRC Laboratory of Medical Sciences"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 6,
  "authors": [
    {
      "name": "V. Dosedelova"
    },
    {
      "name": "J. M. Hawley"
    },
    {
      "name": "E. Melson"
    },
    {
      "name": "L. Schiffer"
    },
    {
      "name": "K.-H. Storbeck"
    },
    {
      "affs": [
        0
      ],
      "name": "W. Arlt"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/1zpr4zcdggz6c.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.10.27.762247",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.10.27.762247v1.full",
  "id": "1zpr4zcdggz6c",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.10.27.762247v1",
  "language": null,
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "biochemistry"
    ]
  ],
  "openalex": null,
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.10.27.762247v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-27",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/1zpr4zcdggz6c.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:30:26Z",
      "native_id": "10.64898/2026.10.27.762247",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.10.27.762247v1"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "biorxiv:biochemistry",
      "subfield": "general"
    }
  ],
  "title": "Optimisation and Validation of a UHPLC-MS/MS Assay Using Timed Post-Column Ammonium Fluoride Infusion for Sensitive Serum Steroids Quantitation",
  "title_text": "Optimisation and Validation of a UHPLC-MS/MS Assay Using Timed Post-Column Ammonium Fluoride Infusion for Sensitive Serum Steroids Quantitation",
  "type": "preprint",
  "updated_at": "2026-10-31T00:20:46Z",
  "url": "https://research.draconitic.com/paper/1zpr4zcdggz6c",
  "venue": "bioRxiv",
  "versions": []
}