Simultaneous determination of cortisol, cortisone, and 6 β-hydroxycortisol in human urine by UPLC with UV detector and application to determine diurnal variations
Author(s)
Author(s)
Xiao. Y.
Hu, M.
Chu, T. T. W.
Fok, B. S. P.
Poon, E. W. M.
Tomlinson, B.
Date Issued
2012
Publisher
Springer
Journal
Chromatographia
Volume
75
Issue
3-4
Start page
169
End page
173
Abstract
In the present studies, a simple rapid ultra performance liquid chromatography (UPLC) method with UV detection for the simultaneous determination of cortisol, cortisone and 6β-hydroxycortisol in human urine was developed. The three analytes and the internal standard dexamethasone were separated on a Waters Acquity UPLC-Tunable UV system with an Acquity UPLC BEH C18 column (50 × 2.1 mm ID, 1.7 μm) using a gradient elution of methanol and water (containing 0.01% formic acid) with a run time of 7 min. The method was accurate and precise, over the ranges of 5–200 ng mL−1 for cortisol, and 10–1,000 ng mL−1 for both cortisone and 6β-hydroxycortisol, and showed good linearity (r 2 > 0.999). This method was applied for the measurement of cortisol, cortisone and 6β-hydroxycortisol in samples collected over different periods as a tool to assess the activity of CYP3A and 11β-hydroxysteroid dehydrogenase type 2 enzymes.
SFU Affiliated Publication
No
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