High-resolution, accurate-mass (HRAM) mass spectrometry urine steroid profiling in the diagnosis of adrenal disorders
Abstract
Abstract BACKGROUND Steroid profiling is a promising diagnostic tool with adrenal tumors, Cushing syndrome (CS), and disorders of steroidogenesis. Our objective was to develop a multiple-steroid assay using liquid-chromatography, high-resolution, accurate-mass mass spectrometry (HRAM LC-MS) and to validate the assay in patients with various adrenal disorders. METHODS We collected 24-h urine samples from 114 controls and 71 patients with adrenal diseases. An HRAM LC-MS method was validated for quantitative analysis of 26 steroid metabolites in hydrolyzed urine samples. Differences in steroid excretion between patients were analyzed based on Z-score deviation from control reference intervals. RESULTS Limits of quantification were 20 ng/mL. Dilution linearity ranged from 80% to 120% with means of 93% to 110% for all but 2 analytes. Intraassay and interassay imprecision ranged from 3% to 18% for all but 1 analyte. Control women had lower excretion of androgen and glucocorticoid precursors/metabolites than men (P < 0.001), but no difference in mineralocorticoids was seen (P = 0.06). Androgens decreased with age in both sexes (P < 0.001). Compared with patients with adrenocortical adenoma (ACA), patients with adrenocortical carcinoma (ACC) had 11 steroids with increased Z scores, especially tetrahydro-11-deoxycortisol (14 vs 0.5, P < 0.001), pregnanetriol (7.5 vs −0.4, P = 0.001), and 5-pregnenetriol (5.4 vs −0.4, P = 0.01). Steroid profiling also demonstrated metabolite abnormalities consistent with enzymatic defects in congenital adrenal hyperplasia and differences in pituitary vs adrenal CS. CONCLUSIONS Our HRAM LC-MS assay successfully quantifies 26 steroids in urine. The statistically significant differences in steroid production of ACC vs ACA, adrenal vs pituitary CS, and in congenital adrenal hyperplasia should allow for improved diagnosis of patients with these diseases.
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Direct biochemical analysis of steroids in blood samples using LC-MS/MS can obviate the specificity problems of immunoassays.
“Direct biochemical analysis of steroids in blood samples using LC-MS/MS can obviate the specificity problems of immunoassays (263-265)”
From Congenital Adrenal Hyperplasia—Current Insights in Pathophysiology, Diagnostics, and Management (2022) · cites this paper as [265]
The use of high-resolution accurate mass (HRAM) mass spectrometry coupled to liquid chromatography systems is being explored as an alternative to traditional MS/MS systems as accurate mass quantification offers the potential to resolve all steroid metabolites with the exception of steroid isomers, unless they are separated chromatographically.
“Moreover, the use of high-resolution accurate mass (HRAM) mass spectrometry coupled to liquid chromatography systems is being explored as an alternative to traditional MS/MS systems as accurate mass quantification offers the potential to resolve all steroid metabolites with the exception of steroid isomers, unless they are separated chromatographically [344].”
From Human steroid biosynthesis, metabolism and excretion are differentially reflected by serum and urine steroid metabolomes: a comprehensive review (2019) · cites this paper as [344]
The urine steroid metabolome of patients with overt Cushing syndrome is characterized by excessive excretion of glucocorticoid and mineralocorticoid precursor metabolites, although androgen metabolites tend to be suppressed in adrenal Cushing but increased in ACTH-dependent Cushing syndrome.
“The urine steroid metabolome of patients with overt Cushing syndrome is characterized by excessive excretion of glucocorticoid and mineralocorticoid precursor metabolites, although androgen metabolites tend to be suppressed (148, 149) in adrenal Cushing but increased in ACTH-dependent Cushing syndrome (150, 151) [Fig. 8A and 8B (1, 148–155)].”
From Steroid Metabolome Analysis in Disorders of Adrenal Steroid Biosynthesis and Metabolism (2019) · cites this paper as [149]
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- Authors
- Hines, J. M., Bancos, I., Bancos, C., Singh, R. D., Avula, A. V., Young, W. F., Grebe, S. K., & Singh, R. J.
- Affiliations
- Mayo Clinic (US)
- Corresponding author
- Singh, Ravinder J
- Venue
- Clinical Chemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 2017-12-01
- Volume
- 63
- Issue
- 12
- Pages
- 1824-1835
- ISSN
- 0009-9147 (ISSN-L)
- 1530-8561
- OpenAlex
- W2749982194
- Cited by
- 111
- References
- 31
- Citations by year
- 2026: 10 · 2025: 7 · 2024: 13 · 2023: 12 · 2022: 12 · 2021: 12 · 2020: 14 · 2019: 17 · 2018: 11 · 2017: 3
- FWCI
- 9.72
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- Clinical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Robert and Elizabeth Strickland Career Development Award
- Center of Individualized Medicine at Mayo Clinic
- Topics
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Health Sciences › Medicine › Surgery › Adrenal and Paraganglionic Tumors
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Hormonal Regulation and Hypertension
- Keywords
- adrenal disorders, Cushing's syndrome, adrenal adenoma, adrenocortical carcinoma, congenital adrenal hyperplasia, adrenal tumors, pregnanetriol, androgens, glucocorticoids, mineralocorticoids, 24-hour urine collection, assay validation
- MeSH terms
- Adrenal Cortex Neoplasms, Adrenal Hyperplasia, Congenital, Adult, Aged, Aged, 80 and over, Chromatography, Liquid, Cushing Syndrome, Female, Humans, Male, Middle Aged, Mass Spectrometry, Steroids, Adrenocortical Adenoma, Adrenocortical Carcinoma, Young Adult, Limit of Detection
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