Adrenal steroids profile by UHPLC-MS/MS in recalled infants of the newborn screening for Congenital Adrenal Hyperplasia

MARÍA GABRIELA BALLERINI; VERÓNICA AMBAO; MARÍA EUGENIA RODRÍGUEZ; MANUELA MARTINEFSKI; ANA VIEYTES; ROSA ENACÁN; MARÍA EUGENIA MONGE; ANA CHIESA; MARÍA GABRIELA ROPELATO

Congreso; 11th International Meeting of Paediatric Endocrinology (IMPE); 2023

Introduction: Recalled newborns in the screening for Congenital Adrenal Hyperplasia (CAH) could be due to analytical steroid interferences. Ultra-high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) constitutes the gold standard for serum steroid quantification.Objective: To describe the serum profile of a panel of 8 adrenal steroids by UHPLC-MS/MS in recalled infants for CAH. Methods: Fifty infants were recalled (32 R-term/18 R-preterm) according to the local algorithm for neonatal CAH screening. CAH due to CYP21 deficiency was ruled-out by clinical follow-up and serum 17OH-Progesterone after solvent extraction. The control group (C) consisted on sixty-five term infants below 2 months of age. Own UHPLC-MS/MS method (Shimadzu Nexera 20A-AB Sciex QTRAP®6500 system), validated against EMA protocols, was used for serum quantification of Pregnenolone (P5), 17OH-Pregnenolone (17OHP5), Dehydroepiandrosterone (DHEA), Progesterone (P4), 17OH-Progesterone (17OHP4), Cortisol (F), Androstenedione (A4), and Testosterone (T). CYP21 and 3B-hydroxysteroid dehydrogenase activities were estimated according to 17OHP4 to cortisol and 5-ene to 4-ene steroids indexes, respectively. Statistics: One-way ANOVA followed by Tukey or linear trend as post-test; Pearson correlation. Results: Excepting for cortisol and T, steroids concentrations were inversely associated to gestational age (r= -0.42, p< 0.01). 17OHP5, DHEA, and A4 were significantly higher in R-term groups as compared to sex-matched controls (p< 0.01; table). Sexual dimorphism was only observed for T concentration (p< 0.0001). Cortisol precursors were always higher in R-preterm infants than in controls and R-term infants.Mean±SEMControl girlsn=34R-Term girlsn=18Control boysn=31R-term boysn=14R-pretermsn=18ng/mLP5 1.2±0.11.3±0.21.2±0.11.8±0.37.5±2.817OHP5 5.0±0.68.1±1.74.9±0.67.2±0.822.3±5.7DHEA 2.7±0.46.2±1.82.3±0.45.4±1.234.5±14.9P4 0.10±0.010.08±0.010.09±0.010.15±0.040.64±0.2717OHP4 0.44±0.040.76±0.100.65±0.070.89±0.153.9±1.30ug/dLF3.6±0.72.7±0.73.5±0.68.1±1.74.8±1.1ng/dLA417±142±523±241±6184±66T3.8±0.310.1±4.4142±1674±1524±7 (girls)43±15 (boys)A trend towards lower 3B-hydroxysteroid dehydrogenase and CYP21 activities were obtained for R-preterm followed by R-term and controls (p< 0.001). Conclusion: The present integrative adrenal steroid panel reveals that cortisol precursors concentration may be higher in recalled term infants as compare to controls, reflecting some degree of postnatal adrenal cortex immaturity on them.