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Clinical Chemistry 38: 346-352, 1992;
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Clinical Chemistry, Vol 38, 346-352, Copyright © 1992 by American Association for Clinical Chemistry

Simultaneous assay of corticosterone and cortisol in plasma by reversed- phase liquid chromatography

M Hariharan, S Naga, T VanNoord and EK Kindt
University of Michigan Medical School, Department of Psychiatry, Ann Arbor 48109-0656.

We have developed a simple, specific, and sensitive reversed-phase liquid-chromatographic method for accurate and simultaneous analysis of corticosterone and cortisol in human plasma. We achieved a detection limit of 300 ng/L for both steroids by modifying the old solid-phase extraction method to make use of "Tef Elutor" C18 columns, using a minibore (100 x 2 mm) analytical column, and using an ultraviolet detector with a 10-mm-pathlength flow cell. With the new extraction method absolute extraction efficiencies were greater than 90% for all the analytes, including the internal standard, flumethasone. The mobile phase was water (containing 5 mL of triethylamine per liter and citric acid to adjust the pH to 6.5), tetrahydrofuran, and acetonitrile (82/10/8 by vol). The average interassay CV for corticosterone at 0-25 micrograms/L was 6.5%; that for cortisol at 0-300 micrograms/L was 3.8%. The analytical recovery relative to the internal standard was 100.2% for cortisol and 102.6% for corticosterone. Possible interferences from drugs and other steroids were studied.


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Copyright © 1992 by the American Association for Clinical Chemistry.