Date of Award

Spring 6-2026

Document Type

Thesis

Degree Name

Master of Science (MS)

Department/Program

Forensic Science

Language

English

First Advisor or Mentor

Marta Concheiro-Guisan

Second Reader

Ana Pego

Third Advisor

Eduardo Geraldo de Campos

Abstract

Chronic stress can lead to long-lasting health problems, such as anxiety, depression, and heart disease. Exposure to stress during pregnancy can have deleterious health effects on the mother, such as shortened gestation, risk of neurodevelopmental disorders, and cardiometabolic disease. Glucocorticoids (cortisol/cortisone) and endocannabinoids (lipid-based neurotransmitters) regulate the human stress response. Hair is an ideal matrix because it allows for evaluation over an extended period of time and is not affected by circadian variations. There were two main goals of this study: 1) develop and validate an analytical method for the determination of stress biomarkers in hair using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and 2) to determine glucocorticoid and endocannabinoid concentrations in hair samples throughout pregnancy. The developed and validated method is as follows: 20 mg of pulverized hair was incubated in 1 mL of methanol:acetonitrile (75:25) in an ultrasound at 55°C for 2h, and analyzed by LC-MS/MS. A total of 66 3-cm-segments across the three trimesters from 35 pregnant women were analyzed. Out of these segments, glucocorticoids were detected in 62 cases (cortisol in 62, cortisone in 54), and endocannabinoids in all of them (OEA and PEA in all, 2-AG in 53, but AEA in none). In 13 cases with segments from all three trimesters, no significant changes for endocannabinoids were observed, while an increase was observed for glucocorticoids. These findings demonstrate that hair analysis enables reliable, longitudinal assessment of glucocorticoids and endocannabinoids during pregnancy, supporting its utility as a non-invasive monitoring tool.

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