Selective LC-MRM/SIM-MS based profiling of adrenal steroids reveals metabolic signatures of 17α-hydroxylase deficiency

Publication date: Available online 31 January 2020Source: The Journal of Steroid Biochemistry and Molecular BiologyAuthor(s): Chaelin Lee, Jung Hee Kim, Sun Joon Moon, Jaeyoon Shim, Hugh I. Kim, Man Ho ChoiABSTRACTAdrenal 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 precisions precision (coefficient of variation) and accuracies 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 &...
Source: The Journal of Steroid Biochemistry and Molecular Biology - Category: Biochemistry Source Type: research