Measurement of Hair Cortisol Throughout Gestation

Background/Objectives: Cortisol has become established as a relevant biomarker due to its association with various pathologies, including its potential utility in mental health research. However, regarding the techniques employed for its analysis, the available literature shows a certain degree of heterogeneity both in the methods used to obtain cortisol and in the analytical techniques employed for its measurement. This makes it difficult to compare results across specific populations, particularly in pregnant women, who experience metabolic and physiological changes characteristic of gestation. Therefore, the aim of this study was to describe the procedure for the extraction and analysis of cortisol in hair samples from pregnant women throughout gestation. Methods: Hair samples, three centimeters in length, were obtained from women during the first, second, and third trimesters of pregnancy. These samples underwent a standardized isopropanol washing step, followed by milling in a laboratory mill using zirconium balls of varying diameters. The resulting hair powder was then weighed and subjected to four incubation cycles using HPLC-grade methanol. Cortisol levels were detected using chemiluminescence immunoassay. Results: Mean hair cortisol levels were 4.1 μg/L (ng/mL) in the first trimester, 11.5 μg/L (ng/mL) in the second trimester, and 6.6 μg/L (ng/mL) in the third trimester. Conclusions: Standardizing the methodology for cortisol extraction improves the reproducibility of results and, in the long term, may support its incorporation into clinical practice as a useful tool for assessing cortisol levels in both pregnant women and the general population, since hair cortisol enables retrospective evaluation of its cumulative exposure over time, approximately on a monthly basis.