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Measurement of Fat-Soluble Vitamins and Antioxidants by HPLC With Electrochemical Array Detection

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Methods in Biological Oxidative Stress

Part of the book series: Methods in Pharmacology and Toxicology ((MIPT))

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Abstract

Fat-soluble vitamins and antioxidants (FSVAs) are a structurally diverse group of compounds (Fig. 1) that play important roles in a wide spectrum of biochemical and physiological processes, e.g., photoreception (vitamin A, retinol); plasma calcium homeostasis (vitamin D2, ergocalciferol; vitamin D3, cholecalciferol); and blood clotting (vitamin K1, phylloquinone). Of considerable interest is the involvement of some FSVAs in oxidative metabolism and the prevention of damage by reactive oxygen species (ROS) (1,2). For example, α-tocopherol (vitamin E) is the primary antioxidant species in the membrane. Here it intercepts lipid peroxyl radicals, thereby inhibiting lipid peroxidation, a self-perpetuating chain reaction, and preventing catastrophic membrane damage. α-Tocopherol is thought to be regenerated from the resulting α-tocopheryl radical by reaction with reduced coenzyme Q10 (CoQ10) (the ubiquinone/ubiquinol system) also located within the membrane, or with cytosolic ascorbic acid (or glutathione) at the cytoplasm-membrane interface (1,2). Another form of vitamin E, γ-tocopherol, readily reacts with reactive nitrogen species (RNS) such as peroxynitrite to form 5-nitro-γ-tocopherol, a marker of RNS production (3).

The chemical structures of some fat-soluble vitamins and antioxidants.

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References

  1. Halliwell, B. and Gutteridge, J. M. C. (1999) Free Radicals in Biology and Medicine. Oxford University Press, Oxford, UK.

    Google Scholar 

  2. Acworth, I. N., McCabe, D. R., and Maher, T. J. (1997) The analysis of free radicals, their reaction products, and antioxidants, in Oxidants, Antioxidants and Free Radicals (Baskin, S. I. and Salem, H., eds.), Washington, DC, Taylor and Francis, pp. 23–77.

    Google Scholar 

  3. Hensley, K., Williamson, K. S., Maidt, M. L., Gabbita, S. P., Grammas, P., and Floyd, R. A. (1999) Determination of biological oxidative stress using high performance liquid chromatography with electrochemical detection. J. High Resol. Chromatogr. 22, 429–437.

    Article  CAS  Google Scholar 

  4. Bui, M. H. (1994) Simple determination of retinol, α-tocopherol and carotenoids (lutein, all-trans-lutein, α-and β-carotene) in human plasma by isocratic liquid chromatography. J. Chromatogr. B 654, 129–133.

    Article  CAS  Google Scholar 

  5. Edlund, P. O. (1988) Determination of coenzyme Q10, a-tocopherol and cholesterol in biological samples by coupled-column liquid chromatography with coulometric and ultraviolet detection. J. Chromatogr. B 425, 87–97.

    Article  CAS  Google Scholar 

  6. Acworth, I. N. and Bowers, M. (1997) An introduction to HPLC-based electrochemical detection: from single electrode to multi-electrode arrays, in Coulometric Electrode Array Detectors for HPLC. Progress in HPLC-HPCE. 6. (Acworth, I. N., Naoi, M., Parvez, H., and Parvez, S., eds.), VSP, The Netherlands, pp. 3–50.

    Google Scholar 

  7. Acworth, I. N., Naoi, M., Parvez, H., and Parvez, S., eds. From single electrode to multi-electrode arrays, in Coulometric Electrode Array Detectors for HPLC. Progress in HPLC-HPCE. 6. (Acworth, I.N., Naoi, M., Parvez, H., and Parvez, S., eds.), VSP, The Netherlands.

    Google Scholar 

  8. Svendsen, C. N. (1993) Multi-electrode array detectors in high-performance liquid chromatography: a new dimension in electrochemical analysis. Analyst 118, 123–129.

    Article  CAS  Google Scholar 

  9. Acworth, I. N., Bailey, B. A., and Maher, T. J. (1998) The use of HPLC with electrochemical detection to monitor reactive oxygen and nitrogen species, markers of oxidative damage and antioxidants: application to the neurosciences, in Neurochemical Markers of Degenerative Nervous Diseases and Drug Addiction. Progress in HPLC-HPCE. 7. (Qureshi, G. A., Parvez, H., Caudy, P., and Parvez, S., eds.), VSP, The Netherlands, pp. 3–56.

    Google Scholar 

  10. Gamache, P. H., Freeto, S. M., and Acworth, I. N. (1999) Coulometric array HPLC analysis of lipid-soluble vitamins and antioxidants. Am. Clin. Lab. 18, 18–19.

    CAS  Google Scholar 

  11. Gamache, P. H., McCabe, D. R., Parvez, H., Parvez, S., and Acworth, I. N. (1997) The measurement of markers of oxidative damage, anti-oxidants and related compounds using HPLC and coulometric array detection, in Coulometric Electrode Array Detectors for HPLC. Progress in HPLC-HPCE. 6. (Acworth, I. N., Naoi, M., Parvez, H., and Parvez, S., eds.), VSP, The Netherlands. pp. 99–126.

    Google Scholar 

  12. Emenhiser, C., Sander, L. C., and Schwartz, S. J. (1995) Capability of a polymeric C30 stationary phase to resolve cis-trans carotenoid isomers in reversedphase liquid chromatography. J. Chromatogr. A 707, 205–216.

    Article  CAS  Google Scholar 

  13. Ferruzzi, M. G., Sander, L. C., Rock, C. L., and Schwartz, S. J. (1998) Caro-tenoid determination in biological microsamples using liquid chromatography with a coulometric electrochemical array. Anal. Biochem. 256, 74–81.

    Article  PubMed  CAS  Google Scholar 

  14. Brown, T. J. and Sharpless, K. E., eds. (1995) Methods for Analysis of Cancer Chemopreventive Agents in Human Serum. NIST Special Publication 874. Washington, DC, US Government Printing Office.

    Google Scholar 

  15. Duewer et al. (1997) NIST/NCI Micronutrients Measurement Quality Assurance Program: Measurement reproducibility, repeatability, stability, and relative accuracy for fat-soluble vitamin-related compounds in human sera. Anal. Chem. 69, 1406–1413.

    Article  PubMed  CAS  Google Scholar 

  16. Duewer et al. (1999) Micronutrients Measurement Quality Assurance Program: Helping participants use interlaboratory comparison exercise results to improve their long-term measurement performance. Anal. Chem. 71, 1870–1878.

    Article  PubMed  CAS  Google Scholar 

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© 2003 Humana Press Inc.,Totowa, NJ

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Gamache, P.H., Ullucci, P.A., Archangelo, J.A., Acworth, I.N. (2003). Measurement of Fat-Soluble Vitamins and Antioxidants by HPLC With Electrochemical Array Detection. In: Hensley, K., Floyd, R.A. (eds) Methods in Biological Oxidative Stress. Methods in Pharmacology and Toxicology. Humana Press. https://doi.org/10.1385/1-59259-424-7:3

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  • DOI: https://doi.org/10.1385/1-59259-424-7:3

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-815-8

  • Online ISBN: 978-1-59259-424-5

  • eBook Packages: Springer Protocols

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