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Targeted LC–MS/MS analysis of steroid glucuronides in human urine
- Source :
- The Journal of Steroid Biochemistry and Molecular Biology. 205:105774
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- Conjugation with glucuronic acid is one of the major metabolic reactions in human steroid hormone catabolism. Recently, increasing interest has been raised concerning the biological roles of steroid glucuronides. We have therefore developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of 15 urinary steroid hormone glucuronides in human urine: androsterone glucuronide (An-G), etiocholanolone glucuronide (Etio-G), epiandrosterone glucuronide (epiAn-G), dihydrotestosterone glucuronide (DHT-G), dehydroepiandrosterone glucuronide (DHEA-G), testosterone glucuronide (T-G), epitestosterone glucuronide (epiT-G), estrone glucuronide (E1-3 G), 17β-estradiol 17-glucuronide (E2-17 G), 17β-estradiol 3-glucuronide (E2-3 G), estriol 16-glucuronide (E3-16 G), pregnenolone glucuronide (Preg-G), tetrahydro-11-deoxycorticosterone 3-glucuronide (THDOC-3 G), cortisol 21-glucuronide (F-G) and pregnanediol glucuronide (PD-G). Sample workup included protein precipitation and solid phase extraction. Internal standards were used to correct for the loss of analytes during sample preparation and analysis. The method showed good linearity (R2≥0.99) and recovery ranged from 89.6 % to 113.8 %. Limit of quantification ranged from 1.9 nmol/L for F-G to 21.4 nmol/L for An-G. Intra-day and inter-day accuracy and precision were below 15 % for all quality controls. The method was successfully applied to 67 urine samples from children and adolescents in whom total concentrations of free and conjugated steroids had been previously determined by GC-MS after enzymatic hydrolysis. Free and sulfated steroids were also measured by LC-MS/MS. In general, the sums of the respective glucuronidated, sulfated and free forms of an analyte corresponded well with its total amount determined after enzymatic hydrolysis by GC-MS. Regarding the most prominent steroid metabolites, the total mean levels of androsterone and etiocholanolone showed an increase up to 5820.0 nmol/L and 4017.8 nmol/L in the group of 15-20 year-old children, respectively. Glucuronide conjugates (4374.3 nmol/L and 3588.5 nmol/L, respectively) dominated. DHEA was excreted mostly as sulfate (0-1 month of age: 184.5 nmol/L; 15-20 years of age: 1618.4 nmol/L) in all age groups. Cortisol was present predominantly as sulfate (mean: 173.8 nmol/L) in newborns. Levels of sulfated cortisol decreased with age, its glucuronidated form increased. The levels of free cortisol were relatively constant throughout childhood. Sex hormones were preferably excreted as glucuronides. In general, steroid hormone metabolites were conjugated to various extents with glucuronic acid or sulfuric acid and their ratio changed over lifetime.
- Subjects :
- Male
0301 basic medicine
Androsterone glucuronide
Endocrinology, Diabetes and Metabolism
Clinical Biochemistry
Dehydroepiandrosterone
Epiandrosterone
Androsterone
Biochemistry
Gas Chromatography-Mass Spectrometry
03 medical and health sciences
chemistry.chemical_compound
Glucuronides
0302 clinical medicine
Endocrinology
Tandem Mass Spectrometry
medicine
Humans
Testosterone
Gonadal Steroid Hormones
Molecular Biology
Chromatography, High Pressure Liquid
Testosterone glucuronide
Etiocholanolone glucuronide
Chromatography
Etiocholanolone
Solid Phase Extraction
Cell Biology
030104 developmental biology
chemistry
030220 oncology & carcinogenesis
Molecular Medicine
Female
Steroids
Glucuronide
Chromatography, Liquid
medicine.drug
Subjects
Details
- ISSN :
- 09600760
- Volume :
- 205
- Database :
- OpenAIRE
- Journal :
- The Journal of Steroid Biochemistry and Molecular Biology
- Accession number :
- edsair.doi.dedup.....822c7e47568bf4626f37cc99ccacc422