OLIVE-DERIVED COMPOSITIONS

Abstract

The present invention relates to a method of producing at least one composition, said method comprising the following steps: (a) preparing a paste from flesh of fermented Kalamon olives; (b) subjecting said paste to a separation process yielding oil, a semisolid fraction, and an aqueous phase, said separation process preferably being centrifugation; and (c) performing one, two or three of the following: (i) harvesting said oil, thereby obtaining a first composition; (ii) drying said semisolid fraction, thereby obtaining a second composition; and (iii) harvesting said aqueous phase, thereby obtaining a third composition.

Claims

1. A method of producing at least one composition, said method comprising: (a) preparing a paste from flesh of fermented Kalamon olives; (b) subjecting said paste to a separation process yielding oil, a semisolid fraction, and an aqueous phase; (c) (i) harvesting said aqueous phase, thereby obtaining a composition A; and (ii) optionally, drying said semisolid fraction, thereby obtaining a composition S.

2. The method of claim 1, further comprising: (ba) washing said semisolid fraction obtained in (b) with water, thereafter combining said water with said aqueous phase obtained in (b), and thereby obtaining a washed and dried composition S of (c)(i), and a combined aqueous phase yielding composition A of (c)(ii).

3. The method of claim 1, further comprising: (d) encapsulating said composition A into a capsule A, wherein (i) said capsule A is a hard capsule; and (ii) an amount of said composition A which is encapsulated in said capsule A corresponds to two to ten of said fermented Kalamon olives.

4. The method of claim 1, further comprising: (e) encapsulating said composition S into a capsule S, wherein (i) said capsule S is a hard capsule or a capsule with an enteric coating; and (ii) an amount of said composition S which is encapsulated in said capsule S corresponds to two to ten of said fermented Kalamon olives.

5. The method of claim 1, further comprising, prior to step (a), allowing fermentation of said olives to occur, wherein said fermentation occurs (i) for a time of about 3 months to about 12 months; and/or (ii) in the presence of autochthonous yeasts.

6. The method of claim 1, wherein said Kalamon olives have been grown in Peloponnese.

7. One or more compositions selected from: (i) composition A; and (ii) composition S, wherein each of said composition A and said composition S is obtained by the method of claim 1.

8. The one or more compositions of claim 7, wherein (i) said composition A comprises phenolic compounds hydroxytyrosol and tyrosol in a mass ratio of 1 mg/g to 50 mg/g; (ii) said capsule A comprises phenolic compounds hydroxytyrosol and tyrosol, in a mass ratio of 1 mg/g to 25 mg/g; and/or (ii) said composition A comprises lactic acid in an amount less than or equal to 150 mg/g.

9. The one or more compositions of claim 7, wherein said composition S comprises triterpenic compounds maslinic acid and oleanolic acid in a mass ratio of 0.1 mg/g to 5 mg/g.

10. A food supplement comprising the one or more compositions of claim 7.

11. A method of treating any one of: (a) dyslipidemia; (b) disorders associated with elevated levels of LDL cholesterol; (c) disorders associated with elevated levels of total cholesterol; and/or (d) disorders associated with low levels of HDL cholesterol, comprising administering the composition of claim 1 to an individual to be treated.

12. The method of claim 11, wherein (a) said levels of LDL cholesterol, total cholesterol and HDL cholesterol are concentrations of LDL cholesterol, total cholesterol and HDL cholesterol in serum, blood or plasma; (b) said elevated levels of LDL cholesterol are serum concentrations; (c) said elevated levels of total cholesterol are serum concentrations above about 200 mg/dl serum; and/or (d) the individual does not receive any further medication for the treatment of hypercholesterolemia.

13. The method of claim 11, wherein said disorders are selected from cardiovascular diseases, atherosclerosis, cardiac infarct, stroke, thrombosis and embolism.

14. (canceled)

15. (canceled)

16. The method of claim 1, wherein said separation process is centrifugation.

17. The method of claim 5, wherein said fermentation occurs for a time of about 4 months to about 6 months.

18. The method of claim 5, wherein said fermentation occurs in the presence of autochthonous yeasts and no further agent triggering fermentation is added.

19. One or more capsules A obtained by the method of claim 3.

20. One or more capsules S obtained by the method of claim 4.

21. A food supplement comprising the one or more capsules of claim 19.

22. A food supplement comprising the one or more capsules of claim 20.

Description

EXAMPLE 1

Manufacture of Compositions of the Invention

[0091] 50 olive fruits are washed and dried gently before weighing. Then, the stones are separated from the flesh and the flesh is weighed.

Weighing Results:

[0092] whole olive fruits: 253.5 gr (average weight of each fruit: 4.71 gr) [0093] 50 olive stones: 44.4 gr [0094] Fruit flesh from 50 olives: 201.0 gr

[0095] After weighing, the flesh is mashed using a blender until an olive paste is obtained. The paste is then centrifuged for 10 min at 4.000 rpm. After centrifugation, the olive paste is found separated into 3 layers (phases), i.e., aqueous phase, lipid phase and a semisolid paste in the middle.

[0096] The upper layer consists of olive oil, which is analyzed using the protocol described in Karkoula et al., Journal of Agricultural and Food Chemistry, 2012, 60, 11696-11170. The total weight is 13.5 gr. Olive oil analysis shows the presence of tyrosol and hydroxytyrosol in relatively low concentrations. “Low concentrations” in this context refers to concentrations relative to those in olive flesh as well as to those in aqueous extracts according to the invention.

[0097] The olive fruit paste stays in the middle layer of the 3-layered material as obtained after the centrifugation. By gently scraping the surface, any olive oil residues that may be present on the upper side of the paste are removed. Then, the paste is stored separately from the liquid phases for further processing as described below. Olive fruit paste after the centrifugation, total weight: 151.3 gr

[0098] The lower phase of the 3-layered material consists of the aqueous phase originally comprised in the olive flesh. Total weight before drying: 22.7 gr The olive fruit paste is divided into 50 mL falcon tubes and equal amount of distilled water to the amount of fruit paste is added in each of them. For resuspending, a heavy duty vortexer is used, for 1 min, until homogenization. The mixture is filtered using paper filter. This step is repeated with fresh water for two more times. All the water from these extractions, including the water from the step of centrifugation are pooled together and filtered once again.

[0099] The combined aqueous phases are dried under vacuum using a rotary evaporator, and then weighed. Total weight of dry extract: 6.25 g. In order to quantify the tyrosol, hydroxyl tyrosol and lactic acid content, we use syringaldehyde as internal standard and qNMR. The NMR spectrum indicates that this extract contains 7 mg/g tyrosol, 9 mg/g hydroxytyrosol and 51 mg/g lactic acid. No maslinic acid is detected.

[0100] To enrich the dried aqueous extract, remove inorganic material (e.g. sodium chloride) and prepare the material for encapsulation, the following procedure is applied: The dry aqueous extract is dissolved in ethanol or isopropanol (1:5 w/v) and the insoluble part is discarded by filtration. The ethanol solution is mixed with microcrystalline cellulose (2:3 w/w dry weight) and dried under vacuum using a rotary evaporator affording a powder that is encapsulated in hard capsules. Each hard capsule has a content of 500 mg including 6 mg tyrosol, 10 mg hydroxytyrosol and 51 mg lactic acid. The preferred daily dose is equivalent to 5 olives is contained in two capsules.

[0101] After the three water extractions as described above, the olive fruit paste weighs 146.3 g. The material is dried using an oven at 60° C. for 72 hours, with gentle stirring every 24 hours. After 3 days the dry weight of the product is 46.5 g. The dry product, similar to a cookie, is pulped using a blender to give a semi-liquid texture resembling a jam or cream. In order to quantify the phenolic content of this material, we use a methanol:water extraction, followed by defatting using cyclohexane and internal standard addition. Using qNMR and data processing, the results show that this product contains 0.24 mg/g tyrosol and 0.54 mg/g hydroxytyrosol, as well as 3.81 mg/g lactic acid and 3.23 mg/g maslinic acid.