Using selected steroids proposals are investigated for coupling liquid chromatographic separations with derivatization after separation for fluorimetric detection. Reverse-phase-, adsorption chromatography, systems with chemically bonded phases as well as one-step reaction with isonicotinoylhydrazide in organic solvents and a multistep reaction of ketosteroids to form dihydrolutidine derivatives in aqueous solution were combined. The effects of the optimized reaction conditions in an air-segmented mixing coil reactor and the mobile phase on fluorescence yield and detection limit are exemplified and discussed. The detection limits for corticosterone are 7 resp. 100 ng in the reactor, when connected to a HPLC-system 100 resp. 500 ng (one-step resp. multi-step reaction).
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Part 2. Reactor Materials and Effects of Variable Dynamic Dimensions
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Schwedt, G., Reh, E. Fluorimetric reaction detectors for high-performance liquid chromatography. Chromatographia 14, 249–253 (1981). https://doi.org/10.1007/BF02260160
- Reaction detector
- Coupling conditions