TY - JOUR
T1 - Selective LC-MRM/SIM-MS based profiling of adrenal steroids reveals metabolic signatures of 17α-hydroxylase deficiency
AU - Lee, Chaelin
AU - Kim, Jung Hee
AU - Moon, Sun Joon
AU - Shim, Jaeyoon
AU - Kim, Hugh I.
AU - Choi, Man Ho
N1 - Funding Information:
This study was supported by a grant from the Korea Institute of Science and Technology Institutional Program (Project No. 2E30480) and the Bio & Medical Technology Development Programs ( NRF-2016M3A9B6902059 ) through the Ministry of Science and ICT. The biospecimens and data used in this study were provided by the Biobank of Seoul National University Hospital, a member of the Korea Biobank Network.
Funding Information:
This study was supported by a grant from the Korea Institute of Science and Technology Institutional Program (Project No. 2E30480) and the Bio & Medical Technology Development Programs (NRF-2016M3A9B6902059) through the Ministry of Science and ICT. The biospecimens and data used in this study were provided by the Biobank of Seoul National University Hospital, a member of the Korea Biobank Network.
Publisher Copyright:
© 2020 Elsevier Ltd
PY - 2020/4
Y1 - 2020/4
N2 - Adrenal steroids are generated in the adrenal cortex and metabolized by various enzymes such as hydroxylases, dehydrogenases, and reductases. Determining the comprehensive metabolic signatures of adrenal steroids can provide insight into their metabolic functions and roles in the pathophysiology of adrenal diseases, including Cushing's syndrome (CS) and congenital adrenal hyperplasia (CAH). To this end, we developed an advanced quantitative profiling method of serum adrenal steroids with liquid chromatography-mass spectrometry (LC–MS) under molecular-specific scan modes. Twenty-seven steroids were separated on a 1.9-μm particle C18 column (50 × 2.1 mm) at a flow rate of 250 μL/min and quantified via triple-quadrupole MS with electrospray ionization. During validation, linearities ( r2) were higher than 0.940 with a limit of quantification of 0.1–5.0 ng/mL, and precision (coefficient of variation) and accuracy (%bias) of 3.7–14.3 % and 96.3–113.1 %, respectively. In contrast with the significantly increased serum levels of mineralocorticoids (P < 0.001), the present LC–MS assay revealed remarkably decreased levels of all glucocorticoids and androgens in a patient diagnosed with 17α-hydroxylase deficiency CAH (P < 0.001) compared to those of age- and sex-matched healthy and CS subjects. In the CAH patient, the metabolic ratios for 17α-hydroxylase were significantly decreased, whereas there was no reduction in the metabolic ratio of 17-hydroxyprogesterone to androstenedione, indicating 17,20-lyase activity. In particular, both pregnenolone and dehydroepiandrosterone sulfates, and their metabolic ratio, were identified as potential biomarkers for 17α-hydroxylase deficiency (all P < 0.001), which were also distinct from those of CS patients. The devised LC–MS assay clearly revealed the metabolic signatures of 17α-hydroxylase deficiency, as a rare phenotype of CAH, compared to both healthy and CS subjects, indicating its utility for screening adrenal diseases.
AB - Adrenal steroids are generated in the adrenal cortex and metabolized by various enzymes such as hydroxylases, dehydrogenases, and reductases. Determining the comprehensive metabolic signatures of adrenal steroids can provide insight into their metabolic functions and roles in the pathophysiology of adrenal diseases, including Cushing's syndrome (CS) and congenital adrenal hyperplasia (CAH). To this end, we developed an advanced quantitative profiling method of serum adrenal steroids with liquid chromatography-mass spectrometry (LC–MS) under molecular-specific scan modes. Twenty-seven steroids were separated on a 1.9-μm particle C18 column (50 × 2.1 mm) at a flow rate of 250 μL/min and quantified via triple-quadrupole MS with electrospray ionization. During validation, linearities ( r2) were higher than 0.940 with a limit of quantification of 0.1–5.0 ng/mL, and precision (coefficient of variation) and accuracy (%bias) of 3.7–14.3 % and 96.3–113.1 %, respectively. In contrast with the significantly increased serum levels of mineralocorticoids (P < 0.001), the present LC–MS assay revealed remarkably decreased levels of all glucocorticoids and androgens in a patient diagnosed with 17α-hydroxylase deficiency CAH (P < 0.001) compared to those of age- and sex-matched healthy and CS subjects. In the CAH patient, the metabolic ratios for 17α-hydroxylase were significantly decreased, whereas there was no reduction in the metabolic ratio of 17-hydroxyprogesterone to androstenedione, indicating 17,20-lyase activity. In particular, both pregnenolone and dehydroepiandrosterone sulfates, and their metabolic ratio, were identified as potential biomarkers for 17α-hydroxylase deficiency (all P < 0.001), which were also distinct from those of CS patients. The devised LC–MS assay clearly revealed the metabolic signatures of 17α-hydroxylase deficiency, as a rare phenotype of CAH, compared to both healthy and CS subjects, indicating its utility for screening adrenal diseases.
KW - 17α-Hydroxylase
KW - Adrenal steroids
KW - Congenital adrenal hyperplasia
KW - Mass spectrometry
UR - http://www.scopus.com/inward/record.url?scp=85079005153&partnerID=8YFLogxK
U2 - 10.1016/j.jsbmb.2020.105615
DO - 10.1016/j.jsbmb.2020.105615
M3 - Article
C2 - 32014605
AN - SCOPUS:85079005153
SN - 0960-0760
VL - 198
JO - Journal of Steroid Biochemistry
JF - Journal of Steroid Biochemistry
M1 - 105615
ER -
