A New HPLC Method for the Simultaneous Analysis of Plasma Retinol, Tocopherols, and Carotenoids and its Application in Cancer Epidemiology

  • Chung S. Yang
  • Kenneth W. Miller
  • Mao-Jung Lee
Chapter
Part of the Experimental Biology and Medicine, Vol. 10 book series (EBAM, volume 10)

Abstract

An isocratic high-performance liquid chromatography (HPLC) method for the simultaneous determination of various fat-soluble vitamins and carotenoids has been developed. The method utilizes a Radial-Pak C18, 8 mm i.d. × 10 cm, 5-µm column and an elution solvent composed of methanol, acetonitrile, and chloroform. Retinol, α-tocopherol, α-carotene, β-carotene, lycopene, zeaxanthin, and two other unidentified carotenoids can be clearly separated and quantified in one HPLC run using α-tocopheryl acetate or tocol as the internal standard. The eluted peaks are quantified by either a photodiode-array detector at preprogrammed wavelengths at the absorption maxima of the compounds or by a dual-wavelength detector at 280 and 436 nm. The total run time is 16 min. With an automatic injector and a programmable detector the system allows unattended operation. The within-run and day-to-day coefficients of variation range from 1 to 8%. In addition, the system can monitor the absorption spectra of the eluent during the HPLC run; this allows the spectral identification of various compounds separated in the same run. The application of this new method in cancer epidemiology was demonstrated in a joint U.S.-China cooperative study on nutrition and esophageal cancer.

Keywords

HPLC Chloroform Lipase Carotenoid Palmitate 

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References

  1. 1.
    Sporn, M. B. and Roberts, A. B. (1983) Cancer Res. 43, 3034–3040.PubMedGoogle Scholar
  2. 2.
    Peto, R., Doll, R., Buckley, J.D., and Sporn, M.B. (1981) Nature 290, 201–208.PubMedCrossRefGoogle Scholar
  3. 3.
    Willett, W.C., Polk, F., Underwood, B.A., Stampfer, M.J., Pressel, S., Rosner, B., Taylor, J.Ö., Schneider, K., and Harnes, C.G. (1984) New Engl.J.Med. 310, 430–434.PubMedCrossRefGoogle Scholar
  4. 4.
    Suberlich, H.E., Dowdy, R.P., and Skala, J.H. (1974) Laboratory tests for the Assessement of Nutrional Status, p. 6, 7 and 76, CRC Press, Boca Raton, FL.Google Scholar
  5. 5.
    Linhares, E.D.R. (1971) Microchem. 16, 467–471.CrossRefGoogle Scholar
  6. 6.
    Arroyave, G., Chichester, C.O., Flores, H., Glover, J., Mejia, L. A., Olson, J. A., Simpson, K. L., and Underwood, B. A. (1982) Biochemical Methodology for the Assessment of Vitamin A Status pp. 60–64. The Nutrition Foundation, Washington, D. C.Google Scholar
  7. 7.
    De Leenheer, A.P., De Bevere, V.O.R.C., De Ruyter, M.G.M., and Claeys, A.E. (1979) J. Chromatogr. 162. 408–413.PubMedCrossRefGoogle Scholar
  8. 8.
    Bieri, J.G., Tolliver, T.J., and Catignani, G.L. (1979) Am J. Clin. Nutr. 32, 2143–2149.PubMedGoogle Scholar
  9. 9.
    Driskell, W.J., Neese, J.W., Bryant, C.C., and Bashor, M.M. (1982) J.Chromatogr. 231, 439–444.PubMedCrossRefGoogle Scholar
  10. 10.
    Broich, C.R., Gerber, L.E., and Erdman, J.W. Jr. (1983) Lipids 18, 253–258.PubMedCrossRefGoogle Scholar
  11. 11.
    Nelis, H.J.C.F. and De Leenheer, A.P. (1983) Anal Chem. 55, 270–275.CrossRefGoogle Scholar
  12. 12.
    Vui11eumier, J.-P., Keller, H.E., Gysel, D., and Hunziker, F. (1983) Int.J. Vit. Nutri. Res. 53, 265–272.Google Scholar
  13. 13.
    Miller, K. W., Lorr, N. A., and Yang, C. S. (1984) Anal. Biocheia. 138, 340–345.CrossRefGoogle Scholar
  14. 14.
    Yang, C. S., Miao, J., Yang, W., Huang, M., Wang, T., Xue, H. You, S., Lu, J., and Wu, J. (1982) Nutr. Cancer 4, 154–164.PubMedCrossRefGoogle Scholar
  15. 15.
    Wi1let, W. C., Stampfer, M., Underwood, B. A., Taylor, J., and Hennekens, C. H. (1983) Am J. Clin. Nutri. 38, 559–566.Google Scholar
  16. 16.
    Yang, C.S., Sun, Y., Yang, Q., Miller, K.W., Li, G., Zheng, S.-F., Ershow, A.G., Blot, W.J., and Li, J. (1984) L. Natl. Cancer Inst. 73, 1449–1453.Google Scholar
  17. 17.
    Ershow, A.G., Zheng, S.-F., Li, G., Li, J., Yang, C.S., and Blot, W.J. (1984) J. Natl. Cancer Inst. 73, 1477–1481.PubMedGoogle Scholar

Copyright information

© The Humana Press Inc. 1986

Authors and Affiliations

  • Chung S. Yang
    • 1
  • Kenneth W. Miller
    • 1
  • Mao-Jung Lee
    • 1
  1. 1.Department of BiochemistryUMDNJ-New Jersey Medical SchoolNewarkUSA

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