Measurement of Hair Cortisol in Healthy Men by Ultra-performance Liquid Chromatography-tandem Mass Spectrometry

Abstract

Published on: October 2022Journal of Young Pharmacists, 2022; 14(4):398-401Original Article | doi:10.5530/jyp.2022.14.80 Authors:Syed Naseeruddin Alvi1,2,*, Sophia De Padua1, Kafa Abuhdeeb1, Muhammad Maher Hammami11Department of Clinical Studies and Empirical Ethics, King Faisal Specialist Hospital and Research Centre Riyadh, KINGDOM OF SAUDI ARABIA.2Environmental Health Program, Department of Cell Biology, King Faisal Specialist Hospital and Research Center, Riyadh, SAUDI ARABIA. Abstract:Objectives: The goal of this study was to use a validated assay to measure hair cortisol levels in healthy persons. Materials and Methods: For quantitative detection of hair-cortisol, a high-performance liquid chromatography-tandem mass spectrometry approach was established and validated. Hair samples were incubated in methanol for 16 hr at 52°C, after which the clear supernatant was sonicated and transfer to a clean tube, where it was dried. The residual sample dissolved in 100 μl methanol containing cortisone (internal standard) and injected. The average extraction recovery was calculated (90.8%). A mobile phase of two mM ammonium acetate (pH 4.2) and acetonitrile (50:50, v:v) was supplied at a flow rate of 0.3 ml/min on an Atlantis dC18 column (2.1 x 100 mm, 3 μm particle size). Multiple reaction monitoring was used in electrospray positive ion mode for mass spectrometry acquisition (m/z: 363.1 → 121.0 and 361.8 → 163.11 for cortisol and cortisone, respectively). Results: Cortisol and internal standard exhibited retention times of about 1.34 and 1.39 min, respectively. In the range of 2-100 ng/ml, the relationship between cortisol level and peak area ratio of cortisol to cortisone was linear, with a detection limit of 0.3 ng/ml. The inter-day coefficient of variation and bias were 7.8% and 11.9%, respectively. 95% of the 88 men had cortisol detectable in their hair samples [mean (SD) length =1.3 (0.5) cm, weight = 33.9 (9.9) mg] and measurable in 49%, with a mean (median) 12.4 (8.7) pg/mg of hair. The method utilized for determining the stability and assessing the level of hair cortisol in samples taken from healthy adult men. Conclusion: For hair cortisol measurements in clinical labs, the described test is reliable, exact, and relevant.Keywords: Cortisol, Cortisone, Hair, Healthy Men, UPLC-MS/MS.