A fluorimetric assay for cortisol |
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Authors: | Daniel?Appel Rolf?D?Schmid Calin-Aurel?Dragan Matthias?Bureik Email author" target="_blank">Vlada?B?UrlacherEmail author |
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Institution: | (1) Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany;(2) Department of Biochemistry, Saarland University, 66123 Saarbrücken, Germany |
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Abstract: | A simple, rapid and sensitive fluorimetric assay for the quantitative determination of cortisol is reported. The assay is
based on the formation of a fluorescent dye when cortisol is incubated with a mixture of sulfuric acid and acetic acid. The
fluorescence spectrum recorded for the resulting dye shows a maximum extinction at 475 nm and a maximum emission at 525 nm.
The solvent 2-methyl-4-pentanone was used for extraction and was found to act as a fluorescence amplifier. A limit of detection
of 2.7 μM was achieved, making it possible to forego solvent evaporation. The assay suffers minor interference from 11-deoxycortisol
which exhibits low fluorescence at λ
ex: 460 nm; λ
em: 505 nm. Typical standard deviations were below 4%. We validated the assay using a biotransformation with recombinant Schizosaccharomyces pombe which regioselectively hydroxylates 11-deoxycortisol to cortisol. The method described herein is suitable for preliminary
screening of microorganisms capable of steroid hydroxylation. |
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Keywords: | Schizosaccharomyces pombe Cortisol 11-deoxycortisol Fluorimetric assay Fission yeast |
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