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Determination of reduced-form glutathione and total glutathione in blood and plasma by high performance liquid chromatography with on-column fluorescence derivatization

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Summary

An on-column fluorometric derivatization method has been developed for the determination of reduced-form glutathione and total glutathione in blood and plasma by high-performance liquid chromatography. Blood hemolysate or diluted plasma was deproteinized with percholoric acid. For total glutathione determination, any oxidized forms of glutathione in samples were converted to reduced-form glutathione with dithiothreitol prior to protein precipitation. Deproteinized samples were injected without pretreatment. Glutathione was fluorescence-derivatized and separated on a C18-bonded vinyl alcohol polymer column withortho-phthalaldehyde in sodium borate buffer-acetonitrile, as mobile phase; theortho-phthalaldehyde-glutathione, fluorescent isoindole adduct is measured by its fluorescence response. Due to the high sensitivity, selectivity reproducibility and the simple HPLC system, the method is particularly suitable for routine assay of numerous or small amounts of samples.

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Authors and Affiliations

  1. Department of Public Health, Kansai Medical University, 570-8506, Moriguchi, Osaka, Japan

    K. Fukunaga & N. Nakazono

  2. Department Biotechnology, Faculty of Engineering, Kansai University, 564-8680, Suita, Osaka, Japan

    M. Yoshida

Authors
  1. K. Fukunaga
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  2. N. Nakazono
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  3. M. Yoshida
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Fukunaga, K., Nakazono, N. & Yoshida, M. Determination of reduced-form glutathione and total glutathione in blood and plasma by high performance liquid chromatography with on-column fluorescence derivatization. Chromatographia 48, 690–694 (1998). https://doi.org/10.1007/BF02467600

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  • DOI: https://doi.org/10.1007/BF02467600

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