| Summary: | Cortisol and its metabolite cortisone have been used as a biological marker of stress in human psychobiological studies. Cortisol and cortisone quantitation may help to examine physiological responses towards stress. A highly sensitive and selective ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method was developed for simultaneous quantitation of cortisol and cortisone in human serum. These two compounds were chromatographically separated using Agilent ZORBAX Eclipse Plus C18 Rapid Resolution HD column (2.1 mm × 50 mm, 1.8 µm) and were eluted with (A) 0.1% formic acid in water and (B) methanol mobile phase. The sample injection volume was 2 µL and the flow rate was set at 0.3 mL/min. The analytes were determined in positive ionization mode and quantitated by multiple-reaction monitoring (MRM) mode. The method was linear from 7.8 to 500 ng/mL for cortisol (r2 = 0.986) and cortisone (r2 = 0.990). The precision, accuracy, and recovery of the method for cortisol ranged from 1.98-10.46%, 90.68-91.94%, 90.38-93.88%, and cortisone ranged from 1.01-11.36%, 93.12-94.79% and 90.48-95.90%, respectively. The developed method has met the quality standards of European Medicines Agency (EMA). © 2019, Malaysian Society of Analytical Sciences. All rights reserved.
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