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Fluorimetric reaction detectors for high-performance liquid chromatography

Part 1. Selection of chemical reactions

Summary

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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References

  1. [1]

    L. R. Snyder, J. Chromatogr.125, 287 (1976).

  2. [2]

    R. S. Deelder, J. Chromatogr.149, 669 (1978).

  3. [3]

    R. Tijssen, Anal. Chim. Acta114, 71 (1980).

  4. [4]

    J. F. K. Huber, K. M. Jonker, H. Poppe, Anal. Chem.52, 2 (1980).

  5. [5]

    R. W. Frei, A. H. M. T. Scholten, J. Chromatogr. Sci.17, 152 (1979).

  6. [6]

    G. Schwedt, Angew. Chem.91, 192 (1979).

  7. [7]

    G. Schwedt, Chemische Reaktionsdetektoren für die schnelle Flüssigkeits-Chromatographie, Heidelberg: Hüthig 1980.

  8. [8]

    L. J. Skeggs, Am. J. Clin. Path.28, 311 (1957).

  9. [9]

    R. S. Deelder, P. J. H. Hendricks, J. Chromatogr.83, 342 (1973).

  10. [10]

    E. Reh, G. Schwedt, Fresenius Z. Anal. Chem.303, 117 (1980).

  11. [11]

    R. Horikawa, T. Tamimura, Z. Tamura, Anal. Biochem.85, 105 (1978).

  12. [12]

    M. Pesez, J. Bartos, Talånta14, 1097 (1967).

  13. [13]

    J. Bartos, M. Pesez, in: Colorimetric and fluorimetric analysis of steroids, S. 40, London-New York-San Francisco: Academic Press 1976.

  14. [14]

    E. Reh, Diplomarbeit Siegen 1979.

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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

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Key Words

  • Reaction detector
  • HPLC
  • Steroids
  • Fluorimetry
  • Coupling conditions