Skip to main content

Selective increase in pinolenic acid (all-cis-5,9,12–18∶3) in Korean pine nut oil by crystallization and its effect on LDL-receptor activity


The aims of this study were to obtain concentrated pinolenic acid (5,9,12–18∶3) from dietary Korean pine (Pinus koraiensis) nut oil by urea complexation and to investigate its cholesteroi-lowering effect on the LDL-receptor activity of human hepatoma HepG2 cells. Pine nut oil was hydrolyzed to provide a low-pinolenic acid-containing FA extract (LPAFAE), followed by crystallization with different ratios of urea in ethanol (EtOH) or methanol (MeOH) as a solvent to produce a high-pinolenic acid-containing FA extract (HPAFAE). The profiles of HPAFAE obtained by urea complexation showed different FA compositions compared with LPAFAE. The long-chain saturated FA palmitic acid (16∶0) and stearic acid (18∶0) were decreased with urea/FA ratios (UFR) of 1∶1 (UFR1), 2∶1 (UFR2), and 3∶1 (UFR3). Linoleic acid (9,12–18∶2) was increased 1.3 times with UFR2 in EtOH, and linolenic acid (9,12,15–18∶3) was increased 1.5 times with UFR3 in MeOH after crystallization. The crystallization with UFR3 in EtOH provided the highest concentration of pinolenic acid, which was elevated by 3.2-fold from 14.1 to 45.1%, whereas that of linoleic acid (9,12–18∶2) was not changed, and that of oleic acid (9–18∶1) was decreased 7.2-fold. Treatment of HepG2 cells with HPAFE resulted in significantly higher internalization of 3,3′-dioctadecylindocarbocyanine-LDL (47.0±0.15) as compared with treatment with LPAFAE (25.6±0.36) (P<0.05). Thus, we demonstrate a method for the concentration of pinolenic acid and suggest that this concentrate may have LDL-lowering properties by enhancing hepatic LDL uptake.

This is a preview of subscription content, access via your institution.





DiI-labeled LDL


high-pinolenic acid-containing FA extract


low-pinolenic acid-containing FA


minimum essential medium


urea/FA ratio


unsaturated polymethylene-interrupted FA


  1. Hollingsworth, P. (1997) Mainstreaming Healthy Foods, Food Technol. 51, 55–58.

    Google Scholar 

  2. Bloch, A., and Thomson, C.A. (1995) Position of the American Dietetic Association: Phytochemicals and Functional Foods, J. Am. Diet. Assoc. 95, 493–496.

    Article  Google Scholar 

  3. Hardy, G. (2000) Nutraceuticals and Functional Foods: Introduction and Meaning, Nutrition 16, 688–689.

    PubMed  CAS  Article  Google Scholar 

  4. Lichtenstein, A.H. (1996) Dietary Fatty Acids and Lipoprotein Metabolism, Curr. Opin. Lipidol. 7, 155–161.

    PubMed  CAS  Article  Google Scholar 

  5. Phillipson, B.E., Rothrock, D.W., Connor, W.E., Harris, W.S., and Illingworth, D.R. (1985) Reduction of Plasma Lipids, Lipoproteins, and Apoproteins by Dietary Fish Oils in Patients with Hypertriglyceridemia, N. Engl. J. Med. 312, 1210–1216.

    PubMed  CAS  Article  Google Scholar 

  6. Imbs, A.B., Nevshupova, N.V., and Pham, L.Q. (1998) Triacylglycerol Composition of Pinus koraiensis Seed Oil, J. Am. Oil Chem. Soc. 75, 865–870.

    CAS  Google Scholar 

  7. Robert, L.W., Eric, D., and Jean-Charles, M. (1997) Positional Distribution of Δ5 Olefinic Acids in Triacylglycerols from Conifer Seed Oils: General and Specific Enrichment in the sn-3 Position, J. Am. Oil Chem. Soc. 74, 515–523.

    Google Scholar 

  8. Asset, G., Staels, B., Wolff, R.L., Bauge, E., Madj, Z., Fruchart, J.C., and Dallongeville, J. (1999) Effects of Pinus pinaster and Pinus koraiensis Seed Oil Supplementation on Lipoprotein Metabolism in the Rat, Lipids 34, 39–44.

    PubMed  CAS  Article  Google Scholar 

  9. Sugano, M., Ikeda, I., Wakamatsu, K., and Oka, T. (1994) Influence of Korean Pine (Pinus koraiensis)-Seed Oil Containing cis-5, cis-9,cis-12-Octadecatrienoic Acid on Polyunsaturated Fatty Acid Metabolism, Eicosanoid Production and Blood Pressure of Rats, Br. J. Nutr. 72, 775–783.

    PubMed  CAS  Article  Google Scholar 

  10. Kurushima, H., Hayashi, K., Toyota, Y., Kambe, M., and Kajiyama, G. (1995) Comparison of Hypocholesterolemic Effects Induced by Dietary Linoleic Acid and Oleic Acid in Hamsters, Atherosclerosis 114, 213–221.

    PubMed  CAS  Article  Google Scholar 

  11. Ibrahim, J.B., and McNamara, D.J. (1988) Cholesterol Homeostasis in Guinea Pigs Fed Saturated and Polyunsaturated Fat Diets, Biochim. Biophys. Acta 963, 109–118.

    PubMed  CAS  Google Scholar 

  12. Gomes, T., and Caponio, F. (1997) Evaluation of the State of Oxidation of Crude Olive-Pomace Oils. Influence of Olive-Pomace Drying and Oil Extraction with Solvent, J. Agric. Food Chem. 45, 1381–1384.

    CAS  Article  Google Scholar 

  13. Ko, S.N., Kim, H., Lee, K.T., Ha, T.Y., Chung, S.H., Lee, S.M., and Kim, I.H. (2003) Optimization of Enzymatic Synthesis of Structured Lipid with Perilla Oil and Medium Chain Fatty Acid, Food Sci. Biotechnol. 12, 253–256.

    CAS  Google Scholar 

  14. Graham, J.M., Higgins, J.A., Gillott, T., Taylor, T., Wilkinson, J., Ford, T., and Billington D. (1996) A Novel Method for the Rapid Separation of Plasma Lipoproteins Using Self-Generating Gradients of Iodixanol, Atherosclerosis 124, 125–135.

    PubMed  CAS  Article  Google Scholar 

  15. Stephan, Z.F., and Yurachek, E.C. (1993) Rapid Fluorometric Assay of LDL Receptor Activity by DiI-Labeled LDL, J. Lipid Res. 34, 325–330.

    PubMed  CAS  Google Scholar 

  16. Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. (1951) Protein Measurement with Folin Phenol Reagent, J. Biol. Chem. 193, 265–275.

    PubMed  CAS  Google Scholar 

  17. Cho, B.H., Dokko, R.C., and Chung, B.H. (2002) Oleic, Linoleic and Linolenic Acids Enhance Receptor-Mediated Uptake of Low Density Lipoproteins in Hep-G2 Cells, J. Nutr. Biochem. 13, 330–336.

    PubMed  CAS  Article  Google Scholar 

  18. Hayes, D.G., Bengtsson, Y.C., Van Alstine, J.M., and Setterwall, F. (1998) Urea Complexation for the Rapid, Ecologically Responsible Fraction of Fatty Acids from Seed Oil, J. Am. Oil Chem. Soc. 75, 1403–1409.

    CAS  Google Scholar 

  19. Grompone, M.A. (1992) Enrichment of Omega-3 PUFAs from Fur Seal Oil, Fat Sci. Technol. 94, 388–394.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations


Corresponding author

Correspondence to Kwang-Won Lee.

About this article

Cite this article

Lee, JW., Lee, KW., Lee, SW. et al. Selective increase in pinolenic acid (all-cis-5,9,12–18∶3) in Korean pine nut oil by crystallization and its effect on LDL-receptor activity. Lipids 39, 383–387 (2004).

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI:


  • EtOH
  • HepG2 Cell
  • Minimum Essential Medium
  • Iodixanol
  • Urea Complexation