High-resolution, accurate-mass (HRAM) mass spectrometry urine steroid profiling in the diagnosis of adrenal disorders

Jolaine M. Hines, Irina Bancos, Cristian Bancos, Raman D. Singh, Aditya V. Avula, William Francis Young, Stefan K. Grebe, Ravinder Jit Singh

Research output: Contribution to journalArticle

30 Citations (Scopus)

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, highresolution, 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.

Original languageEnglish (US)
Pages (from-to)1824-1835
Number of pages12
JournalClinical Chemistry
Volume63
Issue number12
DOIs
StatePublished - Dec 1 2017

Fingerprint

Mass spectrometry
Mass Spectrometry
Steroids
Urine
Metabolites
Adrenocortical Adenoma
Adrenocortical Carcinoma
Congenital Adrenal Hyperplasia
Assays
Cushing Syndrome
Androgens
Pregnanetriol
Pituitary ACTH Hypersecretion
Mineralocorticoids
Glandular and Epithelial Neoplasms
Liquid chromatography
Liquid Chromatography
Glucocorticoids
Dilution
Tumors

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

High-resolution, accurate-mass (HRAM) mass spectrometry urine steroid profiling in the diagnosis of adrenal disorders. / Hines, Jolaine M.; Bancos, Irina; Bancos, Cristian; Singh, Raman D.; Avula, Aditya V.; Young, William Francis; Grebe, Stefan K.; Singh, Ravinder Jit.

In: Clinical Chemistry, Vol. 63, No. 12, 01.12.2017, p. 1824-1835.

Research output: Contribution to journalArticle

Hines, Jolaine M. ; Bancos, Irina ; Bancos, Cristian ; Singh, Raman D. ; Avula, Aditya V. ; Young, William Francis ; Grebe, Stefan K. ; Singh, Ravinder Jit. / High-resolution, accurate-mass (HRAM) mass spectrometry urine steroid profiling in the diagnosis of adrenal disorders. In: Clinical Chemistry. 2017 ; Vol. 63, No. 12. pp. 1824-1835.
@article{bbfc85cf091e43368bd941b01532debb,
title = "High-resolution, accurate-mass (HRAM) mass spectrometry urine steroid profiling in the diagnosis of adrenal disorders",
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, highresolution, 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.",
author = "Hines, {Jolaine M.} and Irina Bancos and Cristian Bancos and Singh, {Raman D.} and Avula, {Aditya V.} and Young, {William Francis} and Grebe, {Stefan K.} and Singh, {Ravinder Jit}",
year = "2017",
month = "12",
day = "1",
doi = "10.1373/clinchem.2017.271106",
language = "English (US)",
volume = "63",
pages = "1824--1835",
journal = "Clinical Chemistry",
issn = "0009-9147",
publisher = "American Association for Clinical Chemistry Inc.",
number = "12",

}

TY - JOUR

T1 - High-resolution, accurate-mass (HRAM) mass spectrometry urine steroid profiling in the diagnosis of adrenal disorders

AU - Hines, Jolaine M.

AU - Bancos, Irina

AU - Bancos, Cristian

AU - Singh, Raman D.

AU - Avula, Aditya V.

AU - Young, William Francis

AU - Grebe, Stefan K.

AU - Singh, Ravinder Jit

PY - 2017/12/1

Y1 - 2017/12/1

N2 - 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, highresolution, 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.

AB - 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, highresolution, 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.

UR - http://www.scopus.com/inward/record.url?scp=85032934524&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85032934524&partnerID=8YFLogxK

U2 - 10.1373/clinchem.2017.271106

DO - 10.1373/clinchem.2017.271106

M3 - Article

C2 - 28814383

AN - SCOPUS:85032934524

VL - 63

SP - 1824

EP - 1835

JO - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

IS - 12

ER -