FRI176 Integrated Profiling Analysis Of Serum Steroids And Catecholamines In A Single Run With LC-MS

Abstract

Abstract Disclosure: J. Noh: None. C. Lee: None. S. Park: None. C. Ahn: None. S. Myung: None. J. Kim: None. M. Choi: None. Adrenal steroids and catecholamines are associated with the response against biochemical stress and adrenal disorders. Metabolic alteration of these hormones could be quantitatively evaluated in diagnostic process, but both adrenal cortex and medullary hormones have been measured in different analytical platforms. To increase the bio-analytical validity, liquid chromatography-mass spectrometry (LC-MS) based simultaneous profiling of 29 steroids and 6 catecholamines was developed. After alkyloxycarbonylation with isobutyl chloroformate, catecholamines and steroids were extracted simultaneously in liquid-liquid extraction. The limit of quantification ranged from 0.1 to 25 ng/mL with extraction recoveries of 78.5-97.0%, while the precision and accuracy were 4.8-7.9% and 98.0-110.9%, respectively. The validated LC-MS assay was then applied to serum samples obtained from 20 individual patients with primary aldosteronism (PA), Cushing’s syndrome (CS) and pheochromocytoma and paraganglioma (PPGL). The significantly increased serum levels of 18-hydroxycorticosterone (p < 0.005), corticosterone (p < 0.05), and 18-hydroxycortisol (p < 0.002) were observed in PA group, while PPGL group showed decreased level of 11-deoxycortisol (p < 0.007). Higher levels of glucocorticoids including 21-deoxycortisol (p < 0.002), 6β-hydroxycortisol (p < 0.001), and tetrahydrocortisone (p < 0.001) and lower levels of androgens including DHEA (p < 0.00001) and DHEA sulfate (p < 0.03) were detected in CS group. Catecholamines, such as norepinephrine (p < 0.0001) and normetanephrine (p < 0.0001), were significantly increased in PPGL patients. The present novel assay was successfully applied to discriminate three different adrenal diseases in a single analytical run. Presentation: Friday, June 16, 2023