Method of obtaining a hydroxytyrosol-rich composition from vegetation water

#This invention relates to a phenolic fraction of a group of compounds present in olive plants known as hydroxytyrosol (3,4-dihydroxyphenylethanol). Particularly, the invention provides an olive extract containing hydroxytyrosol, with low amounts or substantially free of oleoeuropein and tyrosol, and a method of obtaining the same.

#Armstrong, B. K. and Doll, R., International. J. Cancer 15:617-631 (1975). Bartsch, H., et al., Carcinogenesis 20:2209-2218 (1999). Braga, C., et al., Cancer 82:448-453 (1998). Chan, J. M., et al., Seminars in Cancer Biology 8:263-273 (1998). d'Amicis, A. and Farchi, S., in: Advances in Nutrition and Cancer 2 (Zappia, V., et al., Eds.) 67-72, Kluwer Academic/Plenum Publishers, New York (1999). Deiana, M., et al., Free Radic. Biol. Med. 26:762-769 (1999). de la Puerta, R., et al., Biochem. Pharmacol. 57:445-449 (1999). Ficarra, P., et al., Farmaco 46:803-815 (1991). Gerber, M., Epidemiology of Diet and Cancer, ed. M. J. Hill, 263-275 (1994). Kohyama, N., et al., Biosci. Biotechnol. Biochem. 61:347-350 (1997). Kuller, L. H., Journal of the American Dietetic Association 97:S9-S15 (1997). La Vecchia, C., et al., European Journal of Cancer Prevention 7:461-464 (1998). Manna, C., et al., FEBS Letters 470:341-344 (2000). Martin-Moreno, J. M., et al., Int. J. Cancer 58:774-780 (1994). Mattson, F. H. and Grundy, S. M., J. Lipid Res. 26:194-202 (1985). Owen, R. W., et al., J. Can. Res. Clin. Onc. 125:S31 (2000a). Owen, R. W., et al., Eur. J. Cancer 36:1235-1247 (2000b). Owen, R. W., et al., Food Chem. Toxic. 38:647-659 (2000c). Parthasarathy, S., et al., Proc. Natl. Acad. Sci. USA 87:3894-3898 (1990). Petroni, A., et al., Thromb. Res. 78:151-160 (1995). Risch, H. A., et al., Journal of the National Cancer Institute 86:1409-1415 (1994). Romani, A., et al., J. Agric. Food Chem. 47:964-967 (1999). Tsimidou, M., et al., Food Chem. 44:53-60 (1992). Visioli, F., et al., FEBS Letters 468:159-160 (2000). Visioli, F. and Galli, C., Nutr. Rev. 56:142-147 (1998).

#A high amount of dietary fat has been implicated in the development of several diseases (Owen et al., 2000c). Atherosclerosis (Kuller, 1997) and coronary heart disease (Gerber, 1994), as well as cancer of the breast (La Vecchia et al., 1998), prostate (Chan et al., 1998), ovary (Risch et al., 1994), and colon (Armstrong and Doll, 1975) have each been associated with elevated dietary fat. However, evidence indicates that it is not only the amount, but also the type of dietary fat that is important in the etiology of some cancers (Bartsch et al., 1999).

#FIG. 1 shows the structures of phenolic compounds and their precursors detected in olive oil: ligstroside (I); oleuropein glucoside (II); aglycone of ligstroside (III); aglycone of oleuropein glucoside (IV); dialdehydic form of ligstroside aglycone laking a carboxymethyl group (V); dialdehydic form of oleuropein glucoside aglycone lacking a carboxymethyl group (VI); tyrosol (VII); hydroxytyrosol (VIII).

#FIG. 2 shows the HPLC analysis of a hydroxytyrosol-rich composition of the invention after supercritical carbon dioxide extraction from vegetation water.

#FIG. 3 shows the HPLC analysis of a hydroxytyrosol-rich composition of the invention following supercritical carbon dioxide extraction, with synthetic hydroxytyrosol.

#Unless otherwise indicated, all terms used herein have the same meaning as they would to one skilled in the art of the present invention. It is to be understood that this invention is not limited to the particular methodology, protocols, and reagents described, as these may vary.

#By “oleoeuropein” is intended secoiridoid glucoside oleuropein (Structure II in FIG. 1).

#By “tyrosol” is intended 4-hydroxyphenethyl acohol (Structure VII in FIG. 1).