Neonatal reference intervals for salivary steroids: focus on prematurity, stressful events and perinatal betamethasone

Svetlana N Zykova; Anna Plaksienko; Anne Lee Solevåg; Anne Karin Brigtsen; Trond Melbye Michelsen; Ingvil Krarup Sørbye; Mariann Haavik Lysfjord Bentsen; Liv Hanne Bakke; Julie Enge Elboth; Jens Petter Berg; Sandra Rinne Dahl; Per Medbøe Thorsby

doi:10.3389/fped.2026.1808446

Neonatal reference intervals for salivary steroids: focus on prematurity, stressful events and perinatal betamethasone

Front Pediatr. 2026 Jun 16:14:1808446. doi: 10.3389/fped.2026.1808446. eCollection 2026.

Authors

Svetlana N Zykova^#^{1

2}, Anna Plaksienko^#³, Anne Lee Solevåg^{4

5}, Anne Karin Brigtsen⁴, Trond Melbye Michelsen^{5

6}, Ingvil Krarup Sørbye^{5

6}, Mariann Haavik Lysfjord Bentsen⁷, Liv Hanne Bakke¹, Julie Enge Elboth¹, Jens Petter Berg^{5

8}, Sandra Rinne Dahl¹, Per Medbøe Thorsby^{1

5}

Affiliations

¹ Hormone Laboratory, Department of Medical Biochemistry, Endocrinology and Metabolism Research Group, Oslo University Hospital, Oslo, Norway.
² The Public Dental Health Service Competence Centre of Northern Norway, Tromsø, Norway.
³ Oslo Centre of Biostatistics and Epidemiology, University of Oslo, Oslo, Norway.
⁴ Department of Neonatal Intensive Care, Oslo University Hospital, Oslo, Norway.
⁵ Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
⁶ Department of Obstetrics, Oslo University Hospital, Oslo, Norway.
⁷ Department of Paediatrics, Haukeland University Hospital, Bergen, Norway.
⁸ Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.

^# Contributed equally.

Abstract

Context: Painlessly obtained samples can improve diagnostics and care in paediatric conditions with affected steroidogenesis.

Objective: To establish neonatal reference intervals (RIs) and identify sources of biological variation for steroid hormones in saliva.

Methods: Steroid concentrations were determined using LC-MS/MS in 412 saliva samples from 174 infants aged 0-4 weeks, born at gestational age 26-42 weeks, recruited from three neonatal intensive care units and one maternity ward. We used six approaches to construct RIs with partitioning based on results from linear mixed models.

Results: RIs in nmol/L calculated with robust method using median of up to six samples with sufficient volume from each infant were for salivary 17OH-pregnenolone: <4.0; 17OH-progesterone: <2.5; 11-deoxycortisol: 0.02-0.99; 21-deoxycortisol: <0.2; cortisol: 0.65-39; cortisone: 18-154; deoxycorticosterone: <0.04; corticosterone: <0.76; aldosterone: 0.21-4.2; dehydroepiandrosterone: <2.4; dehydroepiandrosterone sulphate: <100; androstenedione: <1.5; testosterone girls: <0.10; testosterone boys: <0.32. Following multivariable adjustment, prematurity was associated with elevated concentrations of mineralocorticoids, androgens and glucocorticoid-precursors (up to 240%, 500% and 1060% respectively), but not cortisol or cortisone. Being on respiratory support was linked with qualitative and quantitative changes in all corticosteroid pathways, while sampling during the first 48 h of life was associated with an increase in adrenocorticotropin-regulated glucocorticoids and androgens (except dehydroepiandrosterone) but not mineralocorticoids. Perinatal exposure to synthetic glucocorticoids led to 33% reduction in salivary cortisol, but not other hormones. Betamethasone was detectable in infants' saliva up to 12 days following maternal administration.

Conclusions: We present neonatal reference intervals for steroid hormones in saliva thereby supporting a non-invasive and painless option for obtaining a steroid hormone profile in this fragile population.

Keywords: 17OH-pregnenolone/17OH-progesterone/11-deoxycortisol/21-deoxycortisol/cortisone/deoxycorticosterone/corticosterone/aldosterone/DHEA/DHEAS/androstenedione/testosterone; adrenal steroid hormones; antenatal corticosteroids; betamethasone; neonatal intensive care unit; preterm/full term infants; reference intervals (RIs); saliva.