WO2021180992A1

WO2021180992A1 - Method for producing oleocanthal

Info

Publication number: WO2021180992A1
Application number: PCT/ES2021/070168
Authority: WO
Inventors: Hristofor MIHO; Antonio FERNÁNDEZ SANTA CRUZ CAMPOS
Original assignee: Santa Cruz Ingeniería, S.L.U.
Priority date: 2020-03-09
Filing date: 2021-03-09
Publication date: 2021-09-16
Also published as: ES2852380A1; ES2852380B2

Abstract

The present invention relates to a method for producing oleocanthal, the oleocanthal obtained using said method, and a composition thereof for use as a food supplement or functional food.

Description

DESCRIPTION

Procedure for obtaining oleocanthal

The present invention refers to a process for obtaining oleocanthal, oleocanthal obtained by said process and a composition thereof for use as a food supplement or as a functional food.

BACKGROUND OF THE INVENTION

The active substance oleocanthal (ligstroside aglycone di aldehyde) is a phenol with properties similar to ibuprofen.

The latest studies have highlighted its anti-inflammatory and antioxidant properties. In addition, in 2011 the European Food Safety Authority (EFSA) considered that the daily consumption of 5 mg of phenolic compounds from olive oil contributes to the oxidative protection of blood lipids. Many clinical trials have shown positive effects against degenerative (Alzheimer's), cardiovascular diseases, cancer diseases, etc. (Beauchamp et al., Nature 2005, 437 (7055), 45-46; Lopez-Huertas & Fonolla, Redox Biology 2017 , 11, 384-389;

Parkinson & Cicerale, Molecules 2016, 21 (12), 1734; Qosa et al., ACS Chemical Neuroscience 2015, 6 (11), 1849-1859; Rallo et al., Scientía Horticulturae 2018, 233, 491-509). Currently, to ingest 5 mg of phenols (including oleocanthal) per day through the consumption of olive oil is practically impossible, since the consumer should ingest a very high amount of oil per day, which, depending on the quality of the oil, would exceed the recommended daily intake of up to 77 grams of fat per day (Liu, Ann G et al. Nutrition Journal 2017, 16:53).

On the other hand, it has been shown that the bioavailability of phenols in the circulatory system (blood) is greatly reduced during digestion. This means that, when a person ingests a quantity of phenols, during the digestion process, these phenolic compounds are degraded, reaching the circulatory system only 4-5% of the quantity ingested (Quintero et al., EurJ Nutr 2018, 57, 1925).

Therefore, it would be desirable to have an efficient procedure for obtaining oleocanthal in high concentrations.

DESCRIPTION OF THE INVENTION One aspect of the present invention refers to the procedure for obtaining oleocanthal that comprises the steps of: i) solid-liquid extraction of the oleocanthal contained in fraction "A" resulting from mixing the pomace and / or alpeorujo from the olive with a solvent to obtain fraction "B" containing the solvent and oleocanthal; and i) isolation and stabilization of the oleocanthal of fraction "B" by adding a vegetable oil combined with gum arabic, and subsequent distillation to obtain fraction "C" to obtain oleo-gelatinous matter enriched in oleocanthal, where the oleocanthal is in a concentration of between 10,000 mg and 80,000 mg of oleocanthal per 1 kg of oleo-gelatinous matter, and preferably in a concentration of 50,000 mg of oleocanthal per 1 kg of oleo-gelatinous matter.

In another embodiment, the invention refers to the process defined above, where step (i) of extraction of fraction "A" is carried out by mixing 1 portion of the olive pomace and / or alpeorujo (solid phase) with 2 portions of the solvent .

In another embodiment, the invention refers to the process defined above, where in step (i) of extraction of phase "A", the solvent is selected from a mixture of ethanol-water, methanol-water or acetonitrile-water, and preferably ethanol-water.

In another embodiment, the invention refers to the process defined above, where the ethanol-water ratio of the solvent from stage (i) of extraction of phase "A" is 80 v / v of ethanol and 20 v / v of water with respect to to the total of the solvent.

In another embodiment, the invention refers to the procedure defined above, where in step (ii) of isolation and stabilization of fraction "B" the vegetable oil is olive oil.

In another embodiment, the invention refers to the process defined above, where fraction "A" is stirred, preferably at between 500 rpm and 3,000 rpm, and more. preferably 2,000 rpm.

In another embodiment, the invention refers to the process defined above, where fraction "A" is stirred for a time between 2 minutes and 30 minutes, preferably for 15 minutes.

In another embodiment, the invention refers to the procedure defined above, where after obtaining fraction "A" in stage (i) and before carrying out stage (ii), a centrifugation stage (iii) is carried out of fraction "A", preferably centrifuging between 5,000 rpm and 20,000 rpm, and more preferably at 15,000 rpm.

In another embodiment, the invention refers to the process defined above, where the distillation of step (ii) of isolation and stabilization of fraction "B" is carried out under vacuum conditions, preferably at 12 psi, and at a temperature of between 10 ° C and 90 ° C, preferably at a temperature of 40 ° C.

Another aspect of the invention relates to oleocanthal obtained by the process defined above. The oleocanthal of the present invention can be administered in the form of any formulation, the nature of which will depend on the nature of the active principle and its route of administration. Preferably, the compounds of the invention can be administered in liquid or encapsulated form.

Thus, in another embodiment, the invention relates to the oleocanthal obtained by the process defined above where the oleocanthal is an oil enriched in oleocanthal or an oleocanthal pill.

In another embodiment, the invention refers to oil enriched in oleocanthal with a concentration of between 1 mg and 10,000 mg of oleocanthal per 1 liter of oil enriched in oleocanthal, and preferably with a concentration of 500 mg of oleocanthal per 1 liter of oil enriched in oleocanthal.

In another embodiment the invention relates to the oleocanthal pill containing between 10 and 600 mg of oleocanthal, preferably containing 50 mg of oleocanthal, and more preferably containing 50 mg of oleocanthal in each 2 gram pill.

Another aspect of the invention refers to the use of oleocanthal as defined above for its use as a food supplement or functional food.

Throughout the present invention, the term "pomace" and / or "pomace" refers to the olive paste obtained during the extraction of olive oil (OA). In particular, the pomace is extracted from the olive oil by means of three-stage centrifuges, and the olive oil is extracted from the olive oil by means of two-stage centrifuges. The pomace and alpeorujo used to obtain oleocanthal are obtained from varieties previously selected for their high concentration in this compound. Examples include, among others, the Alfárara and Pendolino varieties.

Throughout the description and claims the word "comprise" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and characteristics of the invention will emerge in part from the description and in part from the practice of the invention. The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention.

BRIEF DESCRIPTION OF THE FIGURES

Fig. 1 Shows the procedure for extracting the active material (oleocanthal) and obtaining the final product (functional foods and / or food supplements rich in oleocanthal).

EXAMPLES

The invention will be illustrated below by means of tests carried out by the inventors, which show the effectiveness of the product of the invention.

Example 1: Extraction of oleocanthal from olive pomace and selection of olive varieties The extraction of the active material is carried out by means of the solid-liquid extraction process. Specifically, a portion of pomace (solid phase) is mixed with two portions of the solvent ethanol-water (80-20 v / v) - (hereinafter: fraction "A"). Fraction "A" is stirred at 2,000 rpm for 15 minutes. The next step is the centrifugation of said fraction at 15,000 rpm for 5 minutes. This last process enables the separation of the phases: solid phase (pomace) free of phenols and liquid phase (ethanol-water) enriched in phenols. The phase of interest for the process of obtaining the active matter is the liquid phase (hereinafter: fraction "B") which retains all the phenolic compounds (including oleocanthal) previously found naturally in the pomace.

To select the varieties of interest, the “B” fraction of 80 different and independent varieties of olive trees is analyzed through the analysis called “High-performance liquid chromatography (HPLC). Based on these analyzes, the varieties that meet the requirements to obtain the product are selected; which includes a minimum richness of the olive pulp of 30,000 mg / kg of oleocanthal on dry matter. The selected varieties were Kalamon, Plementa Bjelica, Alfafara and Pendolino with a richness in oleocanthal respectively of 42,000, 37,000, 35,400, 31,200 mg / kg on the dry matter of the olive pulp.

Example 2: Isolation and stabilization of oleocanthal in oily phase To isolate and stabilize the oleacanth of fraction "B", a process is carried out that consists of: a) adding 1 portion of refined olive oil containing 12% gum arabic food in 200 portions of fraction "B"; b) distilling fraction "B" at 40 degrees Celsius under vacuum conditions (12 psi) until it is reduced to a gelatinous phase (fraction "C") which retains concentrates from 30,000 to 50,000 mg / kg of oleocanthal.

A) Yes. fraction "C" is obtained by extracting 100 g of dried pomace of the Alfafara variety with 200 ml of the methanol-water solvent mixture in a ratio of 80:20 v / v; After separating the phases and obtaining the aqueous extract (200 ml of the solvent where the phenols were retained), 1 ml of refined oil and 120 mg of gum arabic were added. Next, the hydro-alcoholic phase is evaporated obtaining the oelo-gelatinous phase where the phenolic compounds including oleocanthal were retained.

Subsequently, the oleo-gelatinous phase is re-analyzed to verify that its oleocanthal richness exceeds 30,000 mg / kg. Example 3: Obtaining neocantal oil

One liter of neocantal oil ("Neocantal Oil") was obtained by mixing 1000 ml of commercial extra virgin olive oil with 50 g of fraction "C".

The oil was analyzed using the HPLC method and it was found that its oleocanthal richness reached 500 mg / kg. Example 4: Obtaining Neocantal Pills

One Neocantal pill contains 1 g of fraction "C", equivalent to 50 mg of pure oleocantal. The active material was encapsulated in soft galolatin to camouflage the bitter taste of fraction "C". The total weight of the neocantal pill is 2 grams. After carrying out the analysis of the pill, it was found that its gelatinous core remained intact and stable for 5 months at room temperature and its concentration in oleocanthal was stable at 50,000 mg / kg.

Figure 1 details the procedure for obtaining oleocanthal of the invention, in it the steps indicated refer to: 1- raw material (the pomace of olives previously selected for their richness in oleocanthal; 1a- food ethanol and water ( 80/20 v / v); 2- beating process and obtaining fraction “A”; 3- centrifugation process; 3a- obtaining partially dry pomace (humidity of 20%) free of phenolic compounds; 3b - drying of the pomace up to 10% moisture for use as fertilizer; 4- obtaining the first phenolic extract (fraction “B”); 4th- refined olive oil containing 12% gum arabic; 5- distillation process to concentrate the product and eliminate the alcohol-aquatic phase; 5th- recycling of ethanol; 6- obtaining the phenolic extract in the oleo-gelatinous phase with a richness of up to 50,000 mg / kg in oleocanthal (fraction “C”); 6th- extra virgin olive oil; 7- obtaining oil enriched up to 500 mg / kg in oleocanthal; 6b- process of encapsulation of fraction "C"; 8- Obtaining Oleocanthal Pills with a content of 50 mg of active material (oleocanthal) per liter of olive oil.

Claims

1. Procedure for obtaining oleocanthal that comprises the stages of: i. solid-liquid extraction of the oleocanthal contained in fraction "A" resulting from mixing the olive pomace and / or alpeorujo with a solvent to obtain the fraction "B" that contains the solvent and oleocanthal; and i. isolation and stabilization of oleocanthal from fraction "B" by adding a vegetable oil combined with gum arabic, and subsequent distillation to obtain fraction "C" to obtain oleo-gelatinous matter enriched in oleocanthal, where oleocanthal is in a concentration between 10,000 mg and 80,000 mg of oleocanthal per 1 kg of oleogelatinous matter, and preferably in a concentration of 50,000 mg of oleocanthal per 1 kg of oleo-gelatinous matter.

2. The process according to claim 1, wherein step (i) of extraction of fraction "A" is carried out by mixing 1 portion of the olive pomace and / or alpeorujo (solid phase) with 2 portions of the solvent.

3. The process according to any of claims 1 or 2, wherein in step (i) of extraction of phase "A" the solvent is selected from a mixture of ethanol-water, methanol-water or acetonitrile-water.

4. The process according to claim 3, wherein in step (i) of extraction of phase "A" the solvent is an ethanol-water mixture.

5. The process according to claim 4, wherein the ethanol-water ratio of the solvent from stage (i) of extraction of phase "A" is 80 v / v of ethanol and 20 v / v of water with respect to the total of the solvent.

6. The process according to any of claims 1 to 5, wherein in step (ii) of isolation and stabilization of fraction "B" the vegetable oil is olive oil.

7. The process according to any of claims 1 to 6, wherein fraction "A" is stirred, preferably at between 500 rpm and 3,000 rpm.

8. The process according to any of claims 1 to 7, wherein fraction "A" is stirred for a time between 2 minutes and 30 minutes.

The process according to any of claims 1 to 8, wherein after obtaining fraction "A" in stage (i) and before carrying out stage (ii), a centrifugation stage (iii) is carried out of fraction "A", preferably centrifuging between 5,000 rpm and 20,000 rpm.

10. The process according to any of claims 1 to 9, wherein the distillation of step (ii) of isolation and stabilization of fraction "B" is carried out under vacuum conditions and at a temperature between 10 ° C and 90 ° C.

The process according to claim 10, wherein the distillation of step (ii) of isolation and stabilization of fraction "B" is carried out at a temperature of 40 ° C.

12. The process according to any of claims 10 or 11, wherein the distillation of step (ii) of isolation and stabilization of fraction "B" is carried out under vacuum conditions at 12 psi.

13. Oleocanthal obtained by the process of any of claims 1 to

12.

14. The oleocanthal according to claim 13 wherein the oleocanthal is an oil enriched in oleocanthal or an oleocanthal pill.

15. The oil enriched in oleocanthal according to claim 14 with a concentration of between 1 mg and 10,000 mg of oleocanthal per 1 liter of oil enriched in oleocanthal.

16. The oleocanthal pill according to claim 14 containing between 10 and 600 mg of oleocanthal.

17. Use of the oleocanthal of any of claims 13 to 16, as a food supplement or functional food.