High-performance liquid chromatographic separation and indirect fluorescence detection of thiols |
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Authors: | Pelletier Sarah Lucy Charles A |
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Institution: | Department of Chemistry, University of Alberta, Edmonton, Canada. |
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Abstract: | A fluorescent post-column reaction detection scheme has been devised for selective determination of thiols. The post-column reagent is 40 microM Cd2+ and 100 microM 8-hydroxyquinoline-5-sulfonic acid (HQS) in non-complexing buffer at pH 10. HQS complexes Cd2+ to form a fluorescent product. Thiols in the HPLC effluent compete for complexation of the Cd2+, resulting in a decrease in the fluorescence response. Detection limits of 0.2 microM (0.04 ppm) are achieved for cysteine, homocysteine and glutathione in a 5 min separation. Recoveries from spiked synthetic urine samples are 87-120%. |
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