AN EXTRACT OF PERSEA

Abstract

Suggested is an extract of Persea, in particular Persea americana (Avocado) comprising at least 90 wt.-percent avocado polyols and preferably at least 95 wt.-percent avocadene, obtainable or obtained according to the following steps: (a) providing a source of Persea; (b) subjecting said source of Persea to an extraction step using organic solvents to obtain a first extract; (c) subjecting said first extract to saponification to obtain a second extract; (d) adding a source of calcium to said second extract to precipitate calcium soaps; (e) subjecting the product of step (d) to a separation step to obtain a third extract and said precipitated calcium salts; and optionally (f) subjecting said third extract to purification.

Claims

1. An extract of Persea, in particular Persea americana (Avocado) comprising at least 90 wt.-% avocado polyols and preferably at least 95 wt.-% avocadene, obtainable or obtained according to the following steps: (a) providing a source of Persea; (b) subjecting said source of Persea to an extraction step, preferably using organic solvents to obtain a first extract; (c) subjecting said first extract to saponification to obtain a second extract; (d) adding a source of calcium to said second extract to precipitate calcium soaps; (e) subjecting the product of step (d) to a separation step to obtain a third extract and said precipitated calcium salts; and optionally (f) subjecting said third extract to further purification.

2. A process for obtaining an extract of Persea, in particular Persea americana (Avocado) comprising at least 90 wt.-% avocado polyols and preferably at least 95 wt.-% avocadene, comprising or consisting of the following steps: (a) providing a source of Persea; (b) subjecting said source of Persea to an extraction step preferably using organic solvents to obtain a first extract; (c) subjecting said first extract to saponification to obtain a second extract; (d) adding a source of calcium to said second extract to precipitate calcium soaps; (e) subjecting the product of step (d) to a separation step to obtain a third extract and said precipitated calcium salts; and optionally (f) subjecting said third extract to further purification.

3. A process for obtaining an extract of Persea, in particular Persea americana (Avocado) comprising at least 90 wt.-% avocado polyols and preferably at least 95 wt.-% avocadene, comprising or consisting of the following steps: (a) providing a source of Persea; (b) subjecting said source of Persea to a combined extraction and saponification step using a source of aqueous or alcoholic alkaline compounds to obtain a first extract; (c) adding a source of calcium to said first extract to precipitate calcium soaps; (d) subjecting the product of step (c) to a separation step to obtain a third extract and said precipitated calcium salts; and optionally (e) subjecting said third extract to further purification.

4. The process of claim 2, wherein said source of Persea is either an avocado fruit or a pulp obtained from said avocado fruit.

5. The process of claim 2, wherein the extraction to obtain the first extract is carried out using at least one organic solvent selected from the group consisting of ethanol, n-heptane, n-hexane, cyclohexane, ethyl acetate, butyl acetate, acetone, methylethyl ketone and mixtures thereof.

6. The process of claim 2, wherein the first extract is treated with a source of an alkaline hydroxide optionally as an aqueous or alcoholic solution to obtain a second extract, and the fatty acids present in said second extract are transferred into their soluble alkaline soaps.

7. The process of claim 2, wherein the second extract after saponification has taken place is treated with a calcium salt and said soluble alkaline soaps present in the extract are precipitated as calcium soaps.

8. The process of claim 2, wherein said calcium soaps are separated from the liquid retardant forming the third extract by filtration or centrifugation.

9. The process of claim 8, wherein said third extract is subjected to another extraction step using at least one organic solvent to obtain a fourth extract.

10. The process of claim 8, wherein said calcium soaps, still associated with some liquid, are subjected to a filtration step to obtain the pure calcium soaps and mother liquor.

11. The process of claim 9, wherein said calcium soaps, still associated with some liquid, are subjected to a filtration step to obtain the pure calcium soaps and mother liquor, and said fourth extract and said mother liquor are combined and optionally subjected to further purification.

12. The process of claim 11, wherein (i) said third extract, or (ii) the combined fourth extract and mother liquor, is subjected to a first purification step by MPLC to obtain at least one fraction enriched in avocadene.

13. The process of claim 12, wherein said at least one fraction enriched in avocadene is subjected to HPLC to obtain at least one fraction having at least 95 wt.-percent. avocadene.

14. A method for modulating and/or enhancing the salty taste of an oral composition comprising the following steps: (i) providing an extract of Persea according to claim 1; and (ii) adding a working amount of said extract to a composition intended for oral uptake.

15. (canceled)

16. The process of claim 3, wherein said source of Persea is either an avocado fruit or a pulp obtained from said avocado fruit.

17. A method for modulating and/or enhancing the salty taste of an oral composition comprising the following steps: (i) providing an extract of Persea according to the process of claim 2; and (ii) adding a working amount of said extract to a composition intended for oral uptake.

Description

EXAMPLES

Example 1

[0077] 5 kg of fresh avocados were separated into seed, peel and fruit pulp. 1600 g of the fruit pulp were cut into small pieces and freeze-dried to yield 220 g dried material.

[0078] 100 g of freeze-dried fruit pulp of Persea americana was moulded and extracted successively with 2×350 mL of n-hexane and 2×350 mL of ethyl acetate to give 19.32 g (Extract 1A) and 4.30 g (Extract 1B) of crude extract, respectively. These two extracts were combined on the basis of their TLC profiling and called Extract 1.

[0079] 23 g of Extract 1 was dissolved in a solution of potassium hydroxide in ethyl alcohol (9.2 g/230 mL) and refluxed for five hours (at 80° C.) to achieve Extract 2. 140 mL of distilled water and 63 g of CaCO.sub.3 then were added, respectively, and further heat to reflux for four hours. After that, the hot mixture was cooled to 30° C. at room temperature, and the excess of ethanol reduced by evaporation under reduced pressure. Further distilled water was added to the mixture to optimize the formation of calcium soap.

[0080] The calcium soap precipitated like a gum and was difficult to filtrate. The solid-liquid mixture of Extract 2 was therefore separated by centrifugation. From this separation, the aqueous phase forming Extract 3 was further extracted with ethyl acetate (3×650 mL) to give the enriched avocadene called Extract 4. The gum obtained from the centrifugation was triturated in 350 mL of ethyl acetate and then filtered of by suction to give more Extract 4 (“mother liquor”) and a residual powder consisting of calcium soaps or calcium soaps coated with calcium carbonate.

[0081] For the next investigation the overall process was monitored by Thin Layer Chromatography (TLC); run with an authentic sample of avocadene and a solvent system of CHCl.sub.3—MeOH (94:06).

[0082] Extract 4 (11.10 g) was fractionated on a normal phase MPLC (silica gel eluted with n-Hexane-ethyl acetate 30:70, and pure ethyl acetate) to give three enriched avocadene fractions: Fraction A1 (0.39 g), Fraction A2 (4.10 g) and Fraction A3 (1.35 g). The quantitative analyses and the .sup.1H NMR profile of these fractions showed that each of them was a mixture of avocadene and related metabolites. These fractions were therefore further purified through HPLC.

[0083] HPLC separations of said fractions was carried out in a preparative Kromasil C.sub.18 column (250×50 mm, 10 μm), using water 100% (A) and methanol 100% (B) as eluent, at 80 mL/min flow rate. The solvent system was 72-98% B linear. The system was operated with an evaporative light scattering detector (ELSD) and a photodiode detector (PDA) set at 250 nm to give about 1.200 mg extracts of >98% pure avocadene.

Example 2

[0084] This example shows a process encompassing the steps of [0085] Extraction with aqueous potassium hydroxide [0086] Precipitation [0087] Filtration [0088] Washing [0089] Alcoholic extraction [0090] Concentration/Drying [0091] Solubilisation in ethyl acetate [0092] Crystallisation [0093] Purification by reverse phase chromatography

[0094] 600 g fresh avocado pulp was extracted with 1000 ml 2.5M KOH at 100° C. and a slight overpressure to avoid bumping and foaming for 15 h while stirring vigorously. After cooling down to 20° C. 1,900 ml 1 M CaCl.sub.2 solution was added. The precipitate was collected by filtration using 2 bar pressure and the precipitate was washed using 4000 ml water. The remaining precipitate was extracted at 20° C. with 2,000 ml ethanol to yield an enriched extract with avocado polyols, mainly avocadene and avocadene. The enriched extract was reduced to dryness and solubilized in ethyl acetate. The volume of the ethyl acetate was adjusted to a concentration of 50 g/I avocadene. The resulting clear solution was cooled down slowly (1° C./min) to 0° C. The precipitation started at ca. 5° C. 23 ml solution with a concentration of 50 g/I was obtained and yielded 2.4 g precipitate containing 1.01 g avocadene (by HPLC).

Example 3

[0095] This example shows a process encompassing the steps of [0096] Extraction with aqueous potassium hydroxide [0097] Precipitation [0098] Filtration [0099] Washing [0100] Ethyl acetate extraction [0101] Concentration/Drying [0102] Solubilisation in ethyl acetate [0103] Crystallisation [0104] Purification by reverse phase chromatography

[0105] 600 g fresh avocado pulp was extracted with 1000 ml 2.5M KOH at 100° C. and a slight overpressure to avoid bumping and foaming for 15 h while stirring vigorously. After cooling down to 20° C. 1900 ml 1 M CaCl.sub.2 solution was added. The precipitate was collected by filtration using 2 bar pressure and the precipitate was washed using 4000 ml water. The remaining precipitate was extracted at 20° C. with 2,000 ml ethyl acetate to yield an enriched extract with avocado polyols, mainly avocadene and avocadene. The enriched extract is concentrated under vacuum to yield a concentrated extract containing 50 g/I avocadene. The volume of the ethyl acetate was adjusted to a concentration of 50 g/I avocadene. This clear solution was cooled down slowly (1° C./min) to 0° C. The precipitation started at ca. 5° C. 23 ml solution with a concentration of 50 g/I and yielded 1.2 g precipitate containing 0.52 g avocadene (by HPLC).

Example 4

[0106] This example shows a process encompassing the steps of [0107] Extraction with aqueous potassium hydroxide [0108] Precipitation [0109] Filtration [0110] Washing [0111] Alcoholic extraction [0112] Adjustment of concentration [0113] Purification by reverse phase chromatography

[0114] 600 fresh avocado pulp was extracted with 1000 ml 2.5M KOH at 100° C. and a slight overpressure to avoid bumping and foaming for 15 h while stirring vigorously. After cooling down to 20° C. 1900 ml 1 M CaCl.sub.2 solution was added. The precipitate was collected by filtration using 2 bar pressure and the precipitate was washed using 4000 ml water. The remaining precipitate was extracted at 20° C. with 1,600 ml ethanol to yield an enriched extract with avocado polyols, mainly avocadene and avocadene. The enriched extract was concentrated under vacuum to yield 50 mL of concentrated extract containing 20 g/L avocadene. The concentration of avocadene was adjusted to 10 g/L using water and separated by RP-HPLC yielding 1.08 g of product.