Skip to main content
SpringerLink
Log in

Quantitative determination of total lipid hydroperoxides by a flow injection analysis system

  • Methods
  • Published:
Lipids

Abstract

A flow injection analysis (FIA) system coupled with a fluorescence detection system using diphenyl-1-pyrenylphosphine (DPPP) was developed as a highly sensitive and reproducible quantitative method of total lipid hydroperoxide analysis. Fluorescence analysis of DPPP oxide generated by the reaction of lipid hydroperoxides with DPPP enabled a quantitative determination of the total amount of lipid hydroperoxides. Use of 1-myristoyl-2-(12-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino) dodecanoyl)-sn-glycero-3-phosphocholine as the internal standard improved the sensitivity and reproducibility of the analysis. Several commercially available edible oils, including soybean oil, rapeseed oil, olive oil, corn oil, canola oil, safflower oil, mixed vegetable oils, cod liver oil, and sardine oil were analyzed by the FIA system for the quantitative determination of total lipid hydroperoxides. The minimal amounts of sample oils required were 50 μg of soybean oil (PV=2.71 meq/kg) and 3 mg of sardine oil (PV=0.38 meq/kg) for a single injection. Thus, sensitivity was sufficient for the detection of a small amount and/or low concentration of hydroperoxides in common edible oils. The recovery of sample oils for the FIA system ranged between 87.2±2.6% and 102±5.1% when PV ranged between 0.38 and 58.8 meq/kg. The CV in the analyses of soybean oil (PV=3.25 meq/kg), cod liver oil (PV=6.71 meq/kg), rapeseed oil (PV=12.3 meq/kg), and sardine oil (PV=63.8 meq/kg) were 4.31, 5.66, 8.27, and 11.2%, respectively, demonstrating sufficient reproducibility of the FIA system for the determination of lipid hydroperoxides. The squared correlation (r 2) between the FIA system and the official AOCS iodometric titration method in a linear regression analysis was estimated at 0.9976 within the range of 0.35−77.8 meq/kg of PV (n=42). Thus, the FIA system provided satisfactory detection limits, recovery, and reproducibility. The FIA system was further applied to evaluate changes in the total amounts of lipid hydroperoxides in fish muscle stored on ice.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

DPPP:

diphenyl-1-pyrenylphosphine

FIA:

flow injection analysis

NBD-labeled PC:

1-myristoyl-2(12-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino)dodecanoyl)-sn-glycero-3-phosphocholine

References

  1. Halliwell, B., and Gutteridge, J.M.C. (1999) Free Radicals, Other Reactive Species and Disease, in Free Radicals in Biology and Medicine, 3rd edn. (edited by Halliwell, B., and Gutteridge, J.M.C.), pp. 617–783, Oxford University Press, Oxford.

    Google Scholar 

  2. AOCS (1993) Official and Methods and Recommended Practices of the American Oil Chemists' Society, 4th edn., AOCS Press, Champaign, Method Cd 8–53.

    Google Scholar 

  3. Asakawa, T., and Matsushita, S. (1980) A Colorimetric Microdetermination of Peroxide Values Utilizing Aluminum Chloride as the Catalyst, Lipids 15, 965–967.

    CAS  Google Scholar 

  4. Marshall, P.J., Warso, M.A., and Lands, W.E. (1981) Selective Microdetermination of Lipid Hydroperoxides, Anal. Biochem. 154, 191–199.

    Google Scholar 

  5. Hara, S., Nemoto, K., Yamada, H., and Totani, Y., (1988) HPLC Analysis of Lipid Peroxides, POV Determination of Autoxidation Triglycerides, J. Jpn. Oil Chem. Soc. 37, 541–545.

    CAS  Google Scholar 

  6. Yamada, Y., Terao, J., and Matsushita, S. (1987) Electrochemical Detection of Phospholipid Hydroperoxides in Reverse-Phase High Performance Lipquid Chromatography, Lipids 22, 125–128.

    PubMed  CAS  Google Scholar 

  7. Yamamoto, Y., Brodsky, M.H., Baker, J.C., and Ames, B.N. (1987) Detection and Characterization of Lipid Hydroperoxides at Picomole Levels by High-Performance Liquid Chromatography, Anal. Chem. 160, 7–13.

    CAS  Google Scholar 

  8. Miyazawa, T., Yasuda, K., and Fujimoto, K. (1987) Chemiluminescence High Performance Liquid Chromatography of Phosphatidylcholine Hydroperoxide, Anal. Lett. 20, 915–925.

    CAS  Google Scholar 

  9. Yamamoto, Y., and Niki, E. (1989) Presence of Cholesteryl Ester Hydroperoxide in Human Blood Plasma, Biochem. Biophys. Res. Commun. 165, 989–993.

    Google Scholar 

  10. Miyazawa, T. (1989) Determination of Phospholipid Hydroperoxides in Human Blood Plasma by a Chemiluminescence-HPLC Assay, Free Radic. Biol. Med. 7, 209–217.

    Article  PubMed  CAS  Google Scholar 

  11. Miyazawa, T., Suzuki, T., Fujimoto, K., and Yasuda, K. (1992) Chemiluminescent Simultaneous Determination of Phosphatidylcholine Hydroperoxide and Phosphatidylethanolamine Hydroperoxide in the Liver and Brain of the Rat, J. Lipid Res. 33, 1051–1059.

    PubMed  CAS  Google Scholar 

  12. Chan, H.W.S., and Levett, G. (1977) Autoxidation of Methyl Linoleate. Separation and Analysis of Isomeric Mixtures of Methyl Linoleate Hydroperoxides and Methyl Hydroxylinoleates, Lipids 12, 99–104.

    PubMed  CAS  Google Scholar 

  13. Akasaka, K., Ohrui, H., and Meguro, H. (1988) Fluorometric Determination of Lipid Hydroperoxides with Phosphine Reagents, Nippon Nogei Kagaku Kaishi 62, 1487–1490.

    CAS  Google Scholar 

  14. Akasaka, K., Suzuki, T., Ohrui, H., and Meguro, H. (1987) Study on Aromatic Phosphines for Novel Fluorometry of Hydroperoxides (II). The Determination of Lipid Hydroperoxides with Diphenyl-1-Pyrenylphosphine, Anal. Lett. 20, 797–907.

    CAS  Google Scholar 

  15. Akasaka, K., Ohrui, H., and Meguro, H. (1993) Determination of Triacylglycerol and Cholesterol Ester Hydroperoxides in Human Plasma by High-Performance Liquid Chromatography with Fluorometric Postcolumn Detection, J. Chromatogr. 617, 205–211.

    PubMed  CAS  Google Scholar 

  16. Akasaka, K., Ohrui, H., and Meguro, H. (1994) Measurement of Cholesterol Hydroperoxides of High and Combined Low and Very Low Lipoprotein in Human Plasma, Biosci. Biotech. Biochem. 58, 396–399.

    Article  CAS  Google Scholar 

  17. Akasaka, K., Ohata, A., Ohrui, H., and Meguro, H. (1995) Automatic Determination of Hydroperoxides of Phosphatidylcholine and Phosphatidylethanolamine in Human Plasma, J. Chromatogr. 665, 37–43.

    CAS  Google Scholar 

  18. Kaewsrithong, J., Qiau, D.-F., Ohshima, T., Ushio, H., Yamanaka, H., and Koizumi, C. (2000) Unusual Levels of Phosphatidylcholine Hydroperoxide in Plasma, Red Blood Cell, and Livers of Aromatic Fish, Fisheries Sci. 66, 768–775.

    Article  CAS  Google Scholar 

  19. Kaewsrithong, J., Ohshima, T., and Ushio, H. (2001) Seasonal Variation of Phosphatidylcholine Hydroperoxides in Blood of Sweet Smelt Plecogrossus altivelis, Comp. Biochem. Physiol. 130B, 33–42.

    CAS  Google Scholar 

  20. Kaewsrithong, J., Ohshima, T., Ushio, H., Nagasaka, R., Maita, M., and Sawada, M. (2001) Effects of an Excess Dose of Dietary α-Tocopherol on Hydroperoxide Accumulation and Erythrocyte Osmotic Fragility of Sweet Smelt Plecogrossus altivelis (Temminck et Schlegel), Aquaculture Res. 32, 191–198.

    Article  CAS  Google Scholar 

  21. Ohshima, T., Hopia, A., German, B.J., and Frankel, E.N. (1996) Determination of Hydroperoxides and Structures by High-Performance Liquid Chromatography with Post-column Detection with Diphenyl-1-pyrenylphosphine, Lipids 31, 1091–1096.

    Article  PubMed  CAS  Google Scholar 

  22. Ohshima, T., Ushio, H., and Koizumi, C. (1997) Analysis of Polyunsaturated Fatty Acid Isomers Hydroperoxides by High-Performance Liquid Chromatography with Post-column Fluorescence Detection, in Flavor and Chemistry of Seafoods (Shahidi, F., and Cadwallader, K.R., eds.), ACS Symposium Series 674, pp. 198–217, American Chemical Society, Washington, DC.

    Google Scholar 

  23. Wang, X-H., Ohshima, T., Ushio, H., and Koizumi, C. (1999) Proportion of Geometrical Hydroperoxide Isomers Generated by Radical Oxidation of Methyl Linoleate in Homogeneous Solution and in Aqueous Emulsion, Lipids 34, 675–679.

    Article  PubMed  CAS  Google Scholar 

  24. Wang, X.-H., Ushio, H., and Ohshima, T. (2003) Distributions of Hydroperoxide Positional Isomers Generated by Oxidation of 1-Palmitoyl-2-arachidonyl-sn-glycero-3-phosphocholine in Liposomes and in Methanol Solution, Lipids 38, 65–72.

    Article  PubMed  CAS  Google Scholar 

  25. Bligh, E.G., and Dyer, W.J. (1959) A Rapid Method of Total Lipid Extraction and Purification, Can. J. Biochem. Physiol. 37, 911–917.

    PubMed  CAS  Google Scholar 

  26. Box, G.E.P., Hunter, W.G., and Hunter, J.S. (1978) Statistics for Experimenters, pp. 21–56, John Wiley & Sons, New York.

    Google Scholar 

  27. Akasaka, K., and Ohrui, H. (2000) Development of Phosphine Reagents for the High-Performance Liquid Chromatographic-Fluorometric Determination of Lipid Hydroperoxides, J. Chromatogr. 881, 159–170

    Article  CAS  Google Scholar 

  28. Nakamura, T., Tanaka, R., Higo, Y., Taira, K., and Takeda, T. (1998) Lipid Peroxide Levels in Tissues of Live Fish, Fisheries Sci. 64, 617–620.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Konan 4, Minato-ku, 108-8477, Tokyo, Japan

    Jeong-Ho Sohn, Yusuke Taki, Hideki Ushio & Toshiaki Ohshima

Authors
  1. Jeong-Ho Sohn
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yusuke Taki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hideki Ushio
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Toshiaki Ohshima
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Toshiaki Ohshima.

About this article

Cite this article

Sohn, JH., Taki, Y., Ushio, H. et al. Quantitative determination of total lipid hydroperoxides by a flow injection analysis system. Lipids 40, 203–209 (2005). https://doi.org/10.1007/s11745-005-1376-2

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11745-005-1376-2

Keywords

Search

Navigation