Abstract
Background
Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based assays are employed in more and more clinical laboratories to quantify steroids. The steroid quantification by LC-MS/MS shows great value in screening or diagnosing endocrine disorders; however, the number of functional steroids included in the LC-MS/MS methods is still limited.
Methods
Here, we describe the performance and validation of a 20-steroid plasma panel by LC-MS/MS. The panel included progestogens (including mineralocorticoids and glucocorticoids), androgens and estrogens biosynthesized in steroid metabolic pathways. The LC-MS/MS method was validated according to guidance documents, and subsequently employed to profile steroid changes in endocrine disorders.
Results
Using LC-MS/MS, 20 steroids were separated and quantified in 8 min. Coefficients of variation (CVs) of the 20 analytes at the lower limit of quantification (LLoQ) were all less than 15% (ranging from 1.84% to 14.96%). The linearity of the assay was demonstrated by all the R2 values greater than 0.995. Individual plasma steroids changed significantly in patients with subclinical Cushing’s syndrome (SCS) and polycystic ovary syndrome (PCOS) – 17-hydroxypregnenolone (17-OH-PR), testosterone (T) and dihydrotestosterone (DHT) were significantly decreased in SCS patients, while in PCOS patients, pregnenolone, corticosterone (CORT), androstenedione (A4) and T were significantly increased and DHT was decreased.
Conclusions
The LC-MS/MS method we developed for the quantification of 20 plasma steroids is clinical practicable. The steroid profiling data using this assay indicate its screening value for endocrine disorders. To further explore the value of the assay, more investigations are however needed.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission. W.G. made the concept. W.G., B.W. and Z.W. contributed to the study design. Y.P. and F.C. contributed to method development and method validation. L.Z. and X.L. contributed to classification and collection of clinical samples. J.Q. contributed to data acquisition. Z.W. and H.W. contributed to analysis and interpretation of the data. Z.W. and H.W. contributed to the drafting of the manuscript. H.W., B.P. and W.G. contributed to the critical revision of the manuscript. Q.L. contributed to technology support.
Research funding: Wei Guo was supported by Joint Project of Fudan University & Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, the National Natural Science Foundation of China (81772263, 81972000 and 81572064), Shanghai Municipal Key Clinical Specialty and Key Developing Disciplines of Shanghai Municipal Commission of Health and Family Planning (2015ZB0201). Hao Wang was supported by Shanghai Post-doctoral Excellence Program (2018166) and the project funded by China Postdoctoral Science Foundation (2019M651370, Funder Id: http://dx.doi.org/10.13039/501100002858). Beili Wang was supported by the Sponsorship for the Junior Clinical Medical Technologist in Shanghai (201802), the National Natural Science Foundation of China (81902139) and the Projects from Shanghai Science and Technology Commission (16411952100).
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
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Supplementary Material
The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2019-0869).
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