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Accelerated solvent extraction for quantitative measurement of fatty acids in plasma and erythrocytes

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Lipids

Abstract

Consumption of fish rich in n-3 highly unsaturated FAs (i.e., EPA and DHA) has been suggested to decrease the risk of lifestyle-related diseases such as coronary heart disease, cancer, diabetes, and dementia. Blood eevels of those FA are known appropriate biomarkers of both the corresponding dietary FA intakes and fish consumption. In place of traditional handwork methods for extracting FA, we performed an accelerated solvent extraction (ASE) for at least 13 selected FA in plasma and erythrocytes to measure them by GLC. The FA levels (concentrations and compositions) in 35–50 μL of plasma or erythrocytes were extracted by ASE and measured by GLC. Intra-and interassay coefficients of variation were≤6.0% for both blood materials, except with a minor group of FA (≤1.0% of total FA). When ASE was compared with two traditional handwork methods, FA levels in plasma from 18 healthy subjects were all coincident with very high Pearson's correlation coefficients for the three sets of the sama 18 samples (r≤0.85 to 0.95, P<0.0001), except for 18∶0 (r=0.59, P<0.01). Using ASE and GLC, we have developed a new method for determination the levels of FA in plasma and erythrocytes as biomarkers for dietary intake of fish, fat, and FA. This new method makes it feasible to measure small volumes of samples, automatically, quantitatively, routinely, easily, rapidly and cheaply, with acceptable precision and accuracy.

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Abbreviations

ASE:

accelerated solvent extraction

HUFA:

highly unsaturated FA

THM:

traditional handwork method

References

  1. Egusa, G., Murakami, F., Ito, C., Matsumoto, Y., Kado, S., Okamura, M., Mori, H., Yamane, K., Hara, H., and Yamakido, M. (1993) Westernized Food Habits and Concentrations of Serum Lipids in the Japanese. Arteriosclerosis 100, 249–255.

    Article  CAS  Google Scholar 

  2. Hu, F.B., Stampfer, M.J., Manson, J.E., Ascherio, A., Colditz, G.A., Speizer, F.E., Hennekens, C.H., and Willett, W.C. (1999) Dietary Saturated Fats and Their Food Sources in Relation to the Risk of Coronary Heart Disease in Women. Am. J. Clin. Nutr. 70, 1001–1008.

    PubMed  CAS  Google Scholar 

  3. World Cancer Research Fond/American Institute for Cancer Research. (1997) Food, Nutrition and the Prevention of Cancer: a Global Perspective, pp. 1–670, AICR, Washington, D.C.

    Google Scholar 

  4. Dwyer, J.T. (1997) Human Studies on the Effects of Fatty Acids on Cancer: Summary, Gaps, and Future Research. Am. J. Clin. Nutr. 66 (Suppl 6), 1581S-1586S.

    PubMed  CAS  Google Scholar 

  5. Tsunehara, C.H., Leonetti, D.L., and Fujimoto, W.Y. (1990) Diet of Second-Generation Japanese-American Men With and Without Non-Insulin-Dependent Diabetes, Am. J. Clin. Nutr. 52, 731–738.

    PubMed  CAS  Google Scholar 

  6. Kalmijn, S., Launer, L.J., Ott, A., Witteman, J.C., Hofman, A., and Breteler, M.M. (1997) Dietary Fat Intake and the Risk of Incident Dementia in the Rotterdam Study, Ann. Neurol. 42, 776–782.

    Article  PubMed  CAS  Google Scholar 

  7. Kankaanpaa, P., Sutas, Y., Salminen, S., Lichtenstein, A., and Isolauri, E. (1999) Dietary Fatty Acids and Allergy, Ann. Med. 31, 282–287.

    PubMed  CAS  Google Scholar 

  8. Dolecek, T.A. (1992) Epidemiological Evidence of Relationships Between Dietary Polyunsaturated Fatty Acids and Mortality in the Multiple Risk Factor Intervention Trial, Proc. Soc. Exp. Biol. Med. 200, 177–182.

    PubMed  CAS  Google Scholar 

  9. Zock, P.L., and Katan, M.B. (1998) Linoleic Acid Intake and Cancer Risk: A Review and Meta-Analysis, Am. J. Clin. Nutr. 68, 142–153.

    PubMed  CAS  Google Scholar 

  10. Dyerberg, J., Bang, H.O., and Hjorne, N. (1975) Fatty Acid Composition of the Plasma Lipids in Greenland Eskimos, Am. J. Clin. Nutr. 28, 958–966.

    PubMed  CAS  Google Scholar 

  11. Thorngren, M., and Gustafson, A. (1981) Effects of 11-Week Increases in Dietary Eicosapentaenoic Acid on Bleeding Time, Lipids, and Platelet Aggregation, Lancet 2, 1190–1193.

    Article  PubMed  CAS  Google Scholar 

  12. Kromhout, D., Bosschieter, E.B., and de Lezenne Coulander, C. (1985) The Inverse Relation Between Fish Consumption 20-Year Mortality from Coronary Heart Disease. N. Engl. J. Med. 312, 1205–1209.

    PubMed  CAS  Google Scholar 

  13. Ascherrio, A., Rimm, E.B., Stampfer, M.J., Giovannucci, E.L., and Willett, W.C. (1995) Dietary Intake of Marine n-3 Fatty Acids, Fish Intake, and the Risk of Coronary Disease Among Men, N Engl. J. Med. 332, 977–982.

    Article  Google Scholar 

  14. Daviglus, M.L., Stamler, J., Orencia, A.J., Dyer, A.R., Liu, K., Greenland, P., Walsh, M.K., Morris, D., and Shekelle, R.B. (1997) Fish Consumption and the 30-Year Risk of Fatal Myocardial Infarction. N. Engl. J. Med. 336, 1046–1053.

    Article  PubMed  CAS  Google Scholar 

  15. Fernandez, E., Chatenoud, L., La Vecchia, C., Negri, E., and Franceschi, S. (1999) Fish Consumption and Cancer Risk, Am. J. Clin. Nutr. 70, 85–90.

    PubMed  CAS  Google Scholar 

  16. Cheng, J., Ogawa, K., Kuriki, K., Yokoyama, Y., Kamiya, T., Seno, K., Okuyama, H., Wang, J., Luo, C., Fujii, T., et al., (2003) Increased Intake of n-3 Polyunsaturated Fatty Acids Elevates the Level of Apoptosis in the Normal Sigmoid Colon of Patients Polypectomized for Adenomas/Tumors, Cancer Lett. 193, 17–24.

    Article  PubMed  CAS  Google Scholar 

  17. Mori, T.A., Burke, V., Puddey, I.B., Shaw, J.E., and Beilin, L.J. (2004) Effect of Fish Diets and Weight Loss on Serum Leptin Concentration in Overweight, Treated-Hypertensive Subjects. J. Hypertens. 22, 1983–1990.

    Article  PubMed  CAS  Google Scholar 

  18. Wakai, K., Tamakoshi, K., Date, C., Fukui, M., Suzuki, S., Lin, Y., Niwa, Y., Nishio, K., Yatsuya, H., Kondo, T., et al., for the JACC Study Group. (2005) Dietary Intakes of Fat and Fatty Acids and Risk of Breast Cancer: A Prospective Study in Japan, Cancer Sci. 96, 590–599.

    Article  PubMed  CAS  Google Scholar 

  19. Kojima, M., Wakai, K., Tokudome, S., Suzuki, K., Tamakoshi, K., Watanabe, Y., Kawado, M., Hashimoto, S., Hayakawa, N., Ozasa, K., et al., for the JACC Study Group. (2005) Serum Levels of Polyunsaturated Fatty Acids and Risk of Colorectal Cancer: A Prospective Study, Am. J. Epidemiol. 161, 462–471.

    Article  PubMed  Google Scholar 

  20. Kuriki, K., Wakai, K., Hirose, K. Matsuo, K., Ito, H., Suzuki, T., Saito, T., Kanemitsu, Y., Hirai, T., Kato, T., et al. (in press) Risk of Colorectal Cancer Is Linked to Erythrocyte Compositions of Fatty Acids as Biomarkers for Dietary Intakes of Fish, Fat and Fatty Acids.

  21. Pala, V., Krogh, V., Muti, P., Chajes, V., Riboli, E., Micheli, A., Saadatian, M., Sieri, S., and Berrino, F. (2001) Erythrocyte Membrane Fatty Acids and Subsequent Breast Cancer: A Prospective Italian Study, J. Natl. Cancer Inst. 93, 1088–1095.

    Article  PubMed  CAS  Google Scholar 

  22. Tiemeier, H., van Tuijl, H.R., Hofman, A., Kiliaan, A.J., and Breteler, M.M. (2003) Plasma Fatty Acid Composition and Depression Are Associated in the Elderly: The Rotterdam Study, Am. J. Clin. Nutr. 78, 40–46.

    PubMed  CAS  Google Scholar 

  23. Wang, L., Folsom, A.R., Zheng, Z.J., Pankow, J.S., and Eckfeldt, J.H., for the ARIC Study Investigators. (2003) Plasma Fatty Acid Composition and Incidence of Diabetes in Middle-Aged Adults: The Atherosclerosis Risk in Communities (ARIC) Study, Am. J. Clin. Nutr. 78, 91–98.

    PubMed  CAS  Google Scholar 

  24. Hunter, D. (1990) Biochemical Indicators of Dietary Intake in Nutritional Epidemiology (Willett, W., ed.), pp. 186–216, Oxford University Press, New York.

    Google Scholar 

  25. Lands, W.E.M. (1995) Long-Term Fat Intake and Biomarkers, Am. J. Clin. Nutr. 61 (Suppl), 721S-725S.

    PubMed  CAS  Google Scholar 

  26. Adersen, L.F., Solvoll, K., and Drevon, C.A. (1996) Very-Long-Chain n-3 Fatty Acids as Biomarkers for Intake of Fish and n-3 Fatty Acids Concentrates, Am. J. Clin. Nutr. 64, 301–311.

    Google Scholar 

  27. Arab, L. (2003) Biomarkers of Fat and Fatty Acid Intake, J. Nutr. 133 (Suppl3), 925S-932S.

    PubMed  CAS  Google Scholar 

  28. Kobayashi, M., Sasaki, S., Kawabata, T., Hasegawa, K., Akabane, M., and Tsugane, S. (2001) Single Measurement of Serum Phospholipid Fatty Acid as a Biomarker of Specific Fatty Acid Intake in Middle-Aged Japanese Men, Eur. J. Clin. Nutr. 55, 643–650.

    Article  PubMed  CAS  Google Scholar 

  29. Kuriki, K., Nagaya, T., Imaeda, N., Tokudome, Y., Fujiwara, N., Sato, J., Ikeda, M., Maki, S., and Tokudome, S. (2002) Discrepancies in Dietary Intakes and Plasma Concentrations of Fatty Acids According to Age Among Japanese Female Dietitians, Eur. J. Clin. Nutr. 56, 524–531.

    Article  PubMed  CAS  Google Scholar 

  30. Kuriki, K., Nagaya, T., Tokudome, Y., Imaeda, N., Fujiwara, N., Sato, J., Goto, C., Ikeda, M., Maki, S., Tajima, K., and Tokudome, S. (2003) Plasma Concentrations of (n-3) Highly Unsaturated Fatty Acids Intakes: A Cross-Sectional Study, J. Nutr. 133, 3643–3650.

    PubMed  CAS  Google Scholar 

  31. Wakai, K., Ito, Y., Kojima, M., Tokudome, S., Ozasa, K., Inaba, Y., Yagyu, K., and Tamakoshi, A., JACC Study Group (2005) Intake Frequency of Fish and Serum Levels of Long-Chain n-3 Fatty Acids: A Cross-Sectional Study Within the Japan Collaborative Cohort Study, J. Epidemiol. 15, 211–218.

    Article  PubMed  Google Scholar 

  32. Folch, J., Lees, M., and Sloane-Stanley, G.H. (1957) A Simple Method for the Isolation and Purification of Total Lipides from Animal Tissues, J. Biol. Chem. 226, 497–509.

    PubMed  CAS  Google Scholar 

  33. Skipski, V.P., Good, J.J., Barclay, M., and Reggio, R.B. (1968) Quantitative Analysis of Simple Lipid Classes by Thin-Layer Chromatography. Biochim Biophys Acta. 152, 10–19.

    PubMed  CAS  Google Scholar 

  34. Kim, H.Y., and Salem, N. (1990) Separation of Lipid Classes by Solid Phase Extraction. J. Lipid Res. 31, 2285–2289.

    PubMed  CAS  Google Scholar 

  35. Morrison, W.R., and Smith, L.M. (1964) Preparation of Fatty Acid Methyl Esters and Dimethylacetals from Lipids with Boron Fluoride-Methanol, J. Lipid Res. 5, 600–608.

    PubMed  CAS  Google Scholar 

  36. Lepage, G., and Roy, C.C. (1986) Direct Transesterification of All Classes of Lipids in a One-Step Reaction. J. Lipid Res. 27, 114–120.

    PubMed  CAS  Google Scholar 

  37. Ohta, A., Mayo, M.C., Kramer, N., and Lands, W.E. (1990) Rapid Analysis of Fatty Acids in Plasma Lipids, Lipids 25, 742–747.

    Article  PubMed  CAS  Google Scholar 

  38. Christie, W.W. (1998) Gas Chromatography-Mass Spectrometry Methods for Structural Analysis of Fatty Acids, Lipids 33, 345–353.

    Article  Google Scholar 

  39. Masood, A., Stark, K.D., and Salem, N., Jr. (2005) A Simplified and Efficient Method for the Analysis of Fatty Acid Methyl Esters Suitable for Large Clinical Studies. J. Lipid Res. 46, 2299–2305.

    Article  PubMed  CAS  Google Scholar 

  40. Snyder, J.M., King, J.W., and Jackson, M.A. (1996) Fat Content for Nutritional Labeling by Supercritical Fluid Extraction and an On-Line Lipase Catalyzed Reaction, J. Chromatogr A. 750, 201–207.

    Article  PubMed  CAS  Google Scholar 

  41. Tokudome, S., Yokoyama, Y., Kamiya, T., Seno, K., Okuyama, H., Kuriki, K., Cheng, J., Nakamura, T., Fujii, T., Ichikawa, H., and Itoh, M. (2002) Rationale and Study Design of Dietary Intervention in Patients Polypectomized for Tumors of the Colorectum, Jpn. J. Clin. Oncol. 32, 550–553.

    Article  PubMed  Google Scholar 

  42. Tokudome, S., Imaeda, N., Tokudome, Y. Fujiwara, N., Nagaya, T., Sato, J., Kuriki, K., Ikeda, M., and Maki, S. (2001) Relative Validity of a Semi-Quantitative Food Frequency Questionnaire Versus 28 Day Weighed Diet Records in Japanese Female Dietitians, Eur J Clin Nutr. 55, 735–742.

    Article  PubMed  CAS  Google Scholar 

  43. Tokudome, Y., Kuriki, K., Imaeda, N., Ikeda, M., Nagaya, T., Fujiwara, N., Sato, J., Goto, C., Kikuchi, S., Maki, S., and Tokudome, S. (2003) Seasonal Variation in Consumption and Plasma Concentrations of Fatty Acids in Japanese Female Dietitians, Eur. J. Epidemiol. 18, 945–953.

    Article  PubMed  CAS  Google Scholar 

  44. Jiang, J., Suzuki, S., Xiang, J., Kuriki, K., Hosono, A., Arakawa, K., Wang, J., Nagaya, T., Kojima, M., Katsuda, N., and Tokudome, S. (2005) Plasma Carotenoid, Alpha-Tocopherol and Retinol Concentrations and Risk of Colorectal Adenomas: A Case-Control Study in Japan, Cancer Lett. 226, 133–141.

    Article  PubMed  CAS  Google Scholar 

  45. Tajima, K., Hirose, K., Inoue, M., Takezaki, T., Hamajima, N., and Kuroishi, T. (2000) A Model of Practical Cancer Prevention for Out-Patients Visiting a Hospital: The Hospital-based Epidemiologic Research Program at Aichi Cancer Center (HER-PACC), Asian Pac. J. Cancer Prev. 1, 35–47.

    PubMed  Google Scholar 

  46. Nagaya, Y (1994) Relationships Between Serum Saturated Fatty Acids and Serum Total Cholesterol and HDL-Cholesterol in Humans, Ann. Clin. Biochem. 31, 240–244.

    PubMed  CAS  Google Scholar 

  47. Ozawa, A., Takayanagi, K., Fujita, T., Hirai, A., Hamazaki, T., Terano, T., Tamura, Y., and Kumagai, A. (1982) Determination of Long Chain Fatty Acids in Human Total Plasma Lipids Using Gas Chromatography, Bunseki-kagaku 31, 87–91 (in Japanese).

    CAS  Google Scholar 

  48. Dodge, J.T., Mitchell, C., and Hanahan, D.J. (1963) The Preparation and Chemical Characteristics of Hemoglobin-Free Ghosts of Human Erythrocytes, Arch. Biochem. Biophys. 100, 119–130.

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Kiyonori Kuriki.

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Kuriki, K., Tajima, K. & Tokudome, S. Accelerated solvent extraction for quantitative measurement of fatty acids in plasma and erythrocytes. Lipids 41, 605–614 (2006). https://doi.org/10.1007/s11745-006-5009-6

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  • DOI: https://doi.org/10.1007/s11745-006-5009-6

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