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Journal of Analytical Methods in Chemistry
Volume 2014, Article ID 787483, 6 pages
Research Article

Simultaneous Determination of Cortisol and Cortisone from Human Serum by Liquid Chromatography-Tandem Mass Spectrometry

1Department of Bioanalysis, Seoul Medical Science Institute & Seoul Clinical Laboratories, Seoul 152-766, Republic of Korea
2Department of Laboratory Medicine, Kwandong University College of Medicine, Gangneung 210-701, Republic of Korea
3Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 143-701, Republic of Korea

Received 16 December 2013; Revised 5 February 2014; Accepted 5 February 2014; Published 6 March 2014

Academic Editor: Josep Esteve-Romero

Copyright © 2014 Sanghoo Lee et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


A fast, sensitive, and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was validated and then the levels of cortisol and cortisone from sera of healthy adults were determined by the LC-MS/MS method. One hundred μL of serum sample was directly extracted by adding 2 mL ethyl acetate, followed by chromatographic separation on a C18 column with a mobile phase consisting of 5 mM ammonium acetate and methanol (25 : 75, v/v). The precision, accuracy, and average recovery of the method were 1.5–5.3%, 95.4–102.5%, and 96.4% for cortisol, and 1.9–6.0%, 89.2–98.8%, and 79.9% for cortisone, respectively. The method was linear from 1.0 to 500.0 ng/mL for cortisol and 2.5 to 100.0 ng/mL for cortisone. The limits of detection (LOD) and quantification (LOQ) were 0.2 and 1.0 ng/mL for cortisol, and 1.0 and 2.5 ng/mL for cortisone, respectively. The average cortisol concentration ( ng/mL) of samples collected between 9:00 and 11:00 a.m. was higher approximately 4.4 times than that of cortisone ( ng/mL) . The average cortisone/cortisol ratio was 0.225. Therefore, the LC-MS/MS method may be useful for the diagnosis of some adrenal diseases and the assessment of 11β-hydroxysteroid dehydrogenase (11β-HSD) activity in clinical laboratories.