Cited by 91Analytical Methods in PharmaceuticalsAdrenal Hormones and DisordersClinical Laboratory Practices and Quality Control

Isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of ten steroid metabolites in serum

Søeborg, Tue, Frederiksen, Hanne, Johannsen, Trine Holm, Andersson, Anna-Maria, Juul, Anders·2017·Clinica Chimica Acta

Abstract

An isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of the ten steroid metabolites dehydroepiandrosterone sulfate (DHEAS), progesterone, 17α-hydroxyprogesterone (17-OHP), Δ4-androstenedione (Adione), corticosterone, 11-deoxycortisol, cortisol, cortisone, testosterone (T), and estrone 3-sulfate (E1-S) in serum was developed and validated. Limits of quantification, variability (inter- and intra-day), analytical range and linearity were all found to be acceptable for clinical use. Furthermore, sample stability was evaluated including the influence of freeze-thaw cycles and the effects of temperature and storage time. The method was applied to 391 serum samples from healthy, Danish boys 10-18years old. The concentration ranges of the included steroid metabolites for this population are presented. Concentrations of DHEAS, 17-OHP, Adione and T in the 391 serum samples were furthermore compared to results obtained using an existing LC-MS/MS method in our laboratory. Excellent agreement was found between the methods. Furthermore, the improved sensitivity of the new method allowed for quantification of a number of samples found to be below the LOQs of the existing method. Thus, the two instruments and their associated methods were validated as possible back-ups for each other, which we consider an extremely important issue in high-throughput laboratories analyzing clinical samples on a regular basis. The ten analytes included can be analyzed simultaneously but it is also possible only to include some of these analytes for specific diagnostic purposes which make the new method an extremely useful tool in the clinical laboratory.

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Crossref · 10.1016/j.cca.2017.03.002

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  • Compared to the previous method including ten steroid metabolites, the present method was improved by using an NH4F buffer with a weak basic MeOH buffer and a HESI probe in both positive and negative ionization mode, enabling inclusion of DHT, ALDO, DHEA, 21-DOC, 17-OHPreg and DOC.

    “Compared to our previous method including ten steroid metabolites only [16], the present method was improved by using an NH4F buffer together with a weak basic MeOH buffer as mobile phases on the eluting column and by using a HESI probe operating in both positive and negative ionization mode. Thereby, it was possible to include mainly DHT, ALDO, DHEA, 21-DOC, 17-OHPreg and DOC.”
  • Steroid hormone levels in culture media were measured using a method established to quantify steroid metabolites in serum by isotype-dilution TurboFlow-LC-MS/MS as previously described.

    “Steroid hormone levels were measured using a method established to quantify steroid metabolites in serum by isotype-dilution TurboFlow-LC-MS/MS as previously described (42).”

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Authors
Søeborg, T., Frederiksen, H., Johannsen, T. H., Andersson, A.-M., & Juul, A.
Affiliations
University of Copenhagen (DK)
Corresponding author
Juul, Anders
Venue
Clinica Chimica Acta
Publisher
Elsevier BV
Published
2017-05
Volume
468
Pages
180-186
ISSN
0009-8981 (ISSN-L)
OpenAlex
W2594318512
Cited by
83
References
16
Citations by year
2026: 1 · 2025: 4 · 2024: 22 · 2023: 10 · 2022: 6 · 2021: 11 · 2020: 15 · 2019: 5 · 2018: 8 · 2017: 1
FWCI
5.55
Type
journal article
Language
en
Open access
closed
Locations
  • Clinica Chimica Acta (publishedVersion)
  • PubMed (publishedVersion)
  • Research at the University of Copenhagen (University of Copenhagen) (publishedVersion)
Funders
InterReg (F-22717-07, F-22717-26)
Topics
  • Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
  • Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
Keywords
steroid metabolites, isotope dilution, simultaneous determination, serum, dehydroepiandrosterone sulfate, 17-hydroxyprogesterone, androstenedione, cortisol, corticosterone, 11-deoxycortisol, testosterone, estrone-3-sulfate, method validation, sample stability, pediatric reference values, method comparison
MeSH terms
Adolescent, Blood Chemical Analysis, Child, Chromatography, Liquid, Humans, Isotopes, Male, Steroids, Time Factors, Tandem Mass Spectrometry, Metabolomics

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