Procedure for obtaining an extract of mango seeds, extract thus obtained and uses given to it (Machi

专利摘要:
Procedure for obtaining an extract of mango seeds, extract thus obtained and uses given to it. A method of obtaining an extract of mango seeds by means of a solvent is described, to the extract obtained by said process with antioxidant characteristics and to the uses given thereto in, for example, food preservation. (Machine-translation by Google Translate, not legally binding)
公开号:ES2580761A1
申请号:ES201530241
申请日:2015-02-25
公开日:2016-08-26
发明作者:Ramón CAVA LÓPEZ；Víctor Jesús CANTERO MENA；Luis LADERO GARCÍA
申请人:Universidad de Extremadura；
IPC主号:

专利说明:

Procedure for obtaining an extract of mango seeds, extract thus obtained and uses given to it

The present invention relates to a process for obtaining an extract of mango seeds, to the extract obtained by said procedure and to the uses given thereto. 5

STATE OF THE TECHNIQUE

Several antioxidants used as food protectors are known. Protection against oxidative deterioration of various types of food is a clear concern in the food industry.

As antioxidants, synthetic compounds such as butylhydroxytoluene or butylhydroxyanixol have been used in the food industry. However, there is a growing interest in replacing these synthetic antioxidants with natural antioxidants, which have 15 identical mechanisms of action. The worldwide demand for natural antioxidants has increased due to doubts about food safety. In addition, replacing synthetic additives with natural food additives may have the advantage that the latter have certain health benefits. Most of the natural compounds with antioxidant activity are found in plants and many of them are polyphenolic in nature.

The raw materials of residual origin of agro-industrial processes that can serve as sources of antioxidants deserve particular attention. In this field, some materials such as potato skin residues, 25 olive pomace, alpechines, grape seeds and wine bagasse and grape skins have been considered as raw materials.

Mango (Magnifera indica) is perhaps one of the most known and appreciated exotic fruits. Fleshy fruit whose weight varies from 150g to 2Kg with a somewhat oblong oval shape, obtuse at its ends; Its skin is smooth and thick in color that varies from green, yellow 30 with pink, red or violet tones where it receives direct sunlight. Its flesh is deep yellow, brushing orange. Its bone is fibrous and often penetrates the pulp. Mango is the third tropical fruit in terms of production and import worldwide, immediately located after the tropical banana and pineapple and the fifth of all fruits, with its world production in 2011 being almost 39 million tons. During the transformation of the fruit, a series of residues not used by the food industry are obtained, representing 40% of the total weight of the fruit, approximately 24 thousand tons per year, among which are the shells (8%) and the bones (32%).

With all this, it is of special interest to obtain an extract that encompasses all these characteristics, that is natural, that belongs to an industrialized crop, that is obtained from a part not used within its industry, that has antioxidant activity, which It can be applied in food and that preserves the food in which it is included, of an oxidative deterioration.
 10
DESCRIPTION

The inventors have developed a process for preparing a mango extract with a high polyphenol content which comprises an extraction step with a solvent. The extract obtained with the process of the invention has a high activity as an antioxidant.

Therefore, one aspect of the invention is the process of obtaining a mango seed extract comprising the steps of:
a) crush mango seeds; twenty
b) add the crushed mango seeds obtained in step a) in a solvent selected from: water, acidified water, a C1-6 alcohol or mixtures thereof;
c) stir the mixture of crushed mango seeds and the solvent;
d) separate the two phases formed in step c) and collect the supernatant comprising the extract. 25

The second aspect of the invention relates to an extract of mango seeds obtained by the process of the invention.

Due to the high content of polyphenols comprising the extract obtained by the process of the invention it is suitable for use as an antioxidant. Therefore a third aspect of the invention is the use of the extract of the invention as an antioxidant.

As the origin is natural, the extract of the invention with antioxidant properties is 35
Especially useful for use in a cosmetic or pharmaceutical food product, to promote its preservation. Therefore, a fourth aspect of the invention is a food, cosmetic or pharmaceutical product comprising the extract of the invention.

BRIEF DESCRIPTION OF THE FIGURES 5
FIG. 1 represents mg of malondialdehyde (MDA) / kg of sample, versus days of storage, of the control (black column), of the sample in mango seed extracts being the extraction solvent (water (dark gray column), ethanol ( light gray column) and water: 50:50 v / v ethanol (white column).
DETAILED DESCRIPTION OF THE INVENTION 10

As said, the first aspect of the invention is the process of obtaining a mango seed extract comprising the steps of:
a) crush mango seeds;
b) add the crushed mango seeds obtained in step a) in a solvent selected from: water, acidified water, a C1-6 alcohol and mixtures thereof;
c) stir the mixture of crushed mango seeds and solvent;
d) separate the two phases formed in step c) and collect the supernatant comprising the extract.
 twenty
The first stage is the crushing of the mango seeds until obtaining particles of a size between 0.1mm and 10mm. These seeds can be the byproduct of an industry dedicated to mango or it can be split from the mango itself, in which case the seed will be obtained by manual or mechanical means, as we have said of the mango fruit, after peeling, bone differentiation and pulp, seed extraction after bone fragmentation and removal of the fibrous parts.

The term "C1-6 alcohol" as used herein refers to an alcohol that can have from one carbon to six carbons in a straight or branched chain, with at least one hydroxide function. 30

The term "polyphenols" refers to aromatic compounds comprising at least two hydroxyphenolic groups in the molecule. Flavonoids, a heterogeneous group of plant polyphenols that share a benzopyran structure, are included in this group of compounds. The main flavonoids with antioxidant activity are (+) - catechin, (-) - 35
(-) - epigallocatechin and the corresponding esters of gallic acid, (-) - epicatechin gallate and (-) - epigallocatechin gallate.

The supernatant comprising the mango seed extract may have some impurity and therefore in a preferred embodiment the supernatant obtained in step d) is filtered, in step e).

In a particular embodiment, the process comprises a stage f) of distillation of the supernatant obtained in step d) or after step e) of the filtrate. 10

In a particular embodiment the process comprises a lyophilization step of the supernatant obtained in step d) or the filtrate of step e) or the distillate of step f).

The proportions used of seed: solvent can be modified in order to obtain 15 extracts with greater or lesser concentration and therefore with different antioxidant activity. Preferably the ratio between chopped mango seeds and the solvent is between 1:50 (weight: volume) and 1: 2 (weight: volume). In a particular materialization, the seed: solvent ratio is 1:10 (weight: volume).
 twenty
Likewise, the use of one or the other solvent influences the amount of compounds extracted as well as the type of compounds with antioxidant activity that is obtained due to their characteristics and their affinity for the solvent. Preferably the solvent selected in step c) is a mixture of water and a C1-6 alcohol. In a particular embodiment the solvent selected in step b) is a mixture of water and ethanol.

In a materialization preferably the ratio between water and C1-6 alcohol is between 40:60 v / v and 60:40 v / v.
 30
The conditions of temperature, time, and exposure of the light in step c) can be adjusted in a range in which the obtaining of compounds with antioxidant activity is optimal avoiding degradation. Preferably step c) is carried out at a temperature between 20 ° C and 28 ° C and in dark conditions and for a time of 30 min to 210 min. In a particular materialization the temperature is 25 ° C and the time is two hours.

As said a second aspect of the invention refers to an extract of mango seeds obtained by the process of the invention. In a preferred embodiment, the extract of the invention has a total phenolic compound content between 12 mg Eq gallic acid / g sample and 40 mg Eq gallic acid / g sample. Preferably between 30 and 40 mg Eq gallic acid / g of sample. In a preferred embodiment, the extract of the invention has a content in flavonoid compounds between 2 mg Eq catechin / g sample and 3.5 mg Eq catechin / g sample. Preferably between 2.5 and 3.5 mg Eq catechin / g of sample. 10

As already said, the fourth aspect of the invention is a food, cosmetic or pharmaceutical product comprising the extract of the invention. The application of the extract of the invention is carried out by direct incorporation into the formulation or by dispersion of the extract on the food, cosmetic or pharmaceutical product or any other method that allows the correct distribution of the extract and its antioxidant action. The concentration of extract to be used will be determined by factors specific to the extract (antioxidant capacity) as well as those attributable to the product such as type of product, content and type of fat, concentration in anti and prooxidants, etc. Likewise, the technological processes that may be carried out after the addition of the extract as well as the characteristics of the product must be taken into account.

EXAMPLES

Example 1. Obtaining the extract of the mango seed (Magnifera indica) by using solvent mixture.

To obtain the extract, the fruit was sold from mango sold to the public acquired in a commercial area. With the help of a knife we proceeded to separate the bone from the handle which was fractured to obtain its seed. The seeds obtained were shredded by crushing in a conventional chopper. The extraction of compounds with antioxidant properties was carried out by using two solvents and mixing both. Thus, distilled water, ethanol and a mixture of water: ethanol in 50:50 ratio (vol: vol) were used, which gave rise to three different extracts. The 1:10 ratio (g: mL) of seed and solvent presented the best results in previous experiences, for what was used. The solvent and seed mixtures were kept under stirring for two hours at a controlled temperature of 25 ° C under conditions of reduced luminosity. Once the stirring time elapsed, the phases were separated by centrifugation, obtaining two perfectly differentiated phases. The supernatant phrase was filtered through Whatman No. 54 paper filter and stored in 5 opaque container at refrigeration temperature.

Example 2. Characterization of three extracts of the mango seed (Magnifera indica) by quantifying its total phenolic content.
 10
The quantification of total phenols of mango seed extracts was performed following the procedure described by Singleton et al. (1999). 50 µL of each of the extracts were deposited in a microtiter plate well to which 20 µL of Folin-Cicalteau reagent and 50 µL of 20% sodium carbonate (w / v) were added. The mixture was incubated for one hour at room temperature, after which the absorbance at 765 nm was measured against a blank where the sample was replaced by distilled water. The quantification of the measurements was performed against a standard curve of gallic acid. The results were expressed as equivalents of gallic acid and are reflected in Table 1.

Table 1. Content in total phenolic compounds (mg Eq gallic acid / g of sample) of 20 extracts (water, water: 50:50 v / v ethanol, ethanol) of mango seed.

 Extraction solvent  Total Phenolic Compounds
 Water  13.6 c ± 0.7
 Water: 50:50 v / v ethanol  38.6 to ± 1.2
 Ethanol  27.0 b ± 1.4
p <0.001; N = 8 determinations; a, b, c: In the same column, means with different letters imply statistically significant differences (Tukey's test, p <0.05).
 25
Example 3. Characterization of three extracts of the mango seed (Magnifera indica) by quantifying its flavonoid content.

The quantification of the flavonoid content of the extracts obtained from the mango seed (Magnifera indica) was carried out following the method proposed by Zhishen et al., 30 (1999). This procedure is based on the formation of chelates between AlCl3 and flavonoids in the sample, which have a pink color. Thus, 400 µL of sample was deposited in a test tube to which 60 µL of 10% NaNO2, 60 µL of 20% AlCl3 and 400 µL of 1M NaOH were added. After stirring its absorbance was measured at 510 nm.

The quantification of the measurements carried out was performed against a standard curve of 5 catechin. The results were expressed in mg of equivalent catechin per gram of sample and are reflected in Table 2.

Table 2. Flavonoid content (mg Eq. Catechin / g sample) of extracts (water, water: 50:50 v / v ethanol, ethanol) of mango seed. 10

 Extraction solvent  Phenolic compounds
 Water  2.1 c ± 0
 Water: 50:50 v / v ethanol  3.0 to ± 0.2
 Ethanol  2.4 b ± 0.3

p <0.001; N = 8 determinations; a, b, c: In the same column, means with different letters imply statistically significant differences (Tukey test, p <0.05)
 fifteen
The evaluation of the antioxidant capacity of mango seed extracts was carried out following two synthetic radical reduction methodology, FRAP method (Ferric Reducing Antioxidant Power) and ABTS method (2,2'-azino-bis (3-ethylbenzothiazoline-6 -sulphonic acid), as shown in examples 4 and 5. In these tests, the antioxidant power of the sample is evidenced by changes in the color of a solution that contains a radical that can be reduced. obtained is extrapolated to a calibration curve with Trolox so the results are expressed in TEAC (antioxidant activity equivalent to Trolox).

Example 4. Evaluation of the antioxidant capacity of three mango seed extracts 25 using the ABTS radical

The method proposed by Re et al. (1999). Thus, the ABTS radical was obtained from the equal part mixing of a solution of 7nM ABTS and 2.45nM potassium persulfate. This solution was kept at room temperature for 16 hours in dark conditions for the generation of the radial. The solution was diluted with ethanol to obtain an absorbance of 0.7 (± 0.1) at 734 nm. 245 µL of the solution with the ABTS radical was deposited in a well of a microtiter plate and its absorbance was measured at 734 nm. Then 5 µL of the extract to be tested was added and, after one minute, the measurement was repeated at 734 nm. The results obtained faced a Trolox calibration curve. The results were expressed as mg TEAC / g sample and are reflected in Table 3.

Table 3. Antioxidant capacity measured by the ABTS method (mg TEAC / g sample) of extracts (water, water: 50:50 v / v ethanol, ethanol) of mango seed.
 Extraction solvent  ABTS
 Water  42.2 c ± 2.8
 Water: 50:50 v / v ethanol  56.7 b ± 2.5
 Ethanol  68.9 to ± 3.6
p <0.001; N = 8 determinations; a, b, c: In the same column, means with different letters 10 imply statistically significant differences (Tukey's test, p <0.05).

Example 5. Evaluation of the antioxidant capacity of three mango seed extracts using the FRAP method
 fifteen
This method proposed by Benzie and Strain (1996), determines the capacity of ferric reduction that a sample has at low pH, transforming the tripyridyltriazine complex (TPTZ) with iron (III) to its ferrous form. Thus, the FRAP reagent was prepared from 2.5ml of a 10mM TPTZ solution together with 2.5mL of a 20mM FeCl3-6H2O solution and 25 mL of 0.3mM acetate buffer and pH 3.6. An absorbency reading at 593nm of 20 200mL of the FRAP reagent was made to which 7µL of sample was subsequently added and absorbency measurements were performed after 4 and 30 minutes. The results obtained faced a Trolox calibration curve. The results were expressed as mg TEAC / g sample and are reflected in table 4.
 25
Table 4. Antioxidant capacity measured by the FRAP method (4min and 30min) (mg TEAC / g sample) of extracts (water, water: 50:50 v / v ethanol, ethanol) of mango seed.
 Extraction solvent  FRAP 4min FRAP 30min
 Water  25.1 c ± 2.8 35.2 c ± 1.7
 Water: 50:50 v / v ethanol  56.2 to ± 1.9 82.3 to ± 3.9
 Ethanol  42.5 b ± 2.1 59.0 b ± 3.1
p <0.001; N = 8 determinations; a, b, c: In the same column, means with different letters imply statistically significant differences (Tukey test, p <0.05)

The extract obtained following the provisions of the present invention has shown its effectiveness as an antioxidant agent in the incorporation on food products. 5

The incorporation of three mango seed extracts, obtained by means of three solvents, on a chicken meat product, was effective against oxidative phenomena such as cooking. Example 6 shows the comparative study that evaluated the evolution of the content using the thiobarbituric acid index (TBA-RS) method in a chicken product with added extract versus a product without extract in its formulation.

Example 6. Evaluation of the antioxidant activity of mango seed extracts incorporated in the formulation of a cooked chicken product. fifteen

A chicken product was prepared from breasts obtained in a retail store. The formulation used contained 79% minced meat, 18% water, 1% salt and 2% of each of the extracts with respect to the final weight. The extracts of water, ethanol and water: ethanol (50:50 v / v) and a control without added extract were used. The mixture was distributed in plastic tubes with approximately 30g in each tube. The tubes were introduced in hot water until a temperature in the center of the mixture of 72oC was obtained and kept for 30min to obtain a cooked product. Subsequently, the samples were cooled on ice and stored at 4 ° C for 8 days.
 25
The quantification of oxidative processes was performed using the thiobarbituric acid index (TBA-RS) method described by Salih et al. (1987) that measures the content of certain secondary products derived from lipid oxidation such as malondialdehyde (MDA). Thus, 2g of sample was taken and homogenized with 7.5mL of 3.86% perchloric acid. 2mL of homogenate were mixed with 2mL of 0.02M TBA and the mixture was heated 30 to 90 ° C for 30 minutes. Subsequently, an aliquot was taken to measure its absorbency at 532 nm in front of a target. The results obtained faced a calibration curve of 1,1,3,3-tetraethoxypropane (PET). The results were expressed as mg of MDA / Kg sample and are reflected in Figure 1.

权利要求:
Claims (15)
[1]

1. Procedure for obtaining an extract of mango seeds comprising the steps of:
a) crush mango seeds; 5
b) add the crushed mango seeds obtained in step a) in a solvent selected from: water, acidified water, a C1-6 alcohol and mixtures thereof;
c) stir the mixture of crushed mango seeds and solvent;
d) separate the two phases formed in step c) and collect the supernatant comprising the extract. 10

[2]
2. Method according to claim 1, wherein the supernatant obtained in step d) is filtered in step e).

[3]
3. Method according to claims 1-2, which comprises a stage f) of distillation of the supernatant obtained in step d) or after step e) of the filtrate.

[4]
4. The method according to claims 1-3, which comprises a lyophilization step g) of the supernatant obtained in step d) or the filtrate of step e) or the distillate of step f).
 twenty
[5]
5. Method according to claims 1-4 wherein in step a) the mango seeds are crushed to obtain a particle size between 0.1mm and 10mm.

[6]
6. Method according to claims 1-5 wherein the ratio between crushed mango seeds and the solvent is between 1:50 (weight: volume) and 1: 2 (weight: volume). 25

[7]
7. Method according to claims 1-6 wherein the solvent selected in step b) is a mixture of water and a C1-6 alcohol.

[8]
8. Method according to claim 7 wherein the ratio between water and C1-6 alcohol is between 40:60 v / v and 60:40 v / v.

[9]
9. Process according to claims 7-8 wherein the solvent selected in step b) is a mixture of water and ethanol.
 35
[10]
10. Method according to claims 1-6 wherein the stirring stage c) is carried out at a temperature between 20 ° C and 28 ° C, in dark conditions and for a time of 30 min to 210 min.

[11]
11. Mango seed extract obtained by the procedure described in 5 claims 1-10

[12]
12. Mango seed extract according to claim 11, which has a total phenolic content between 12 and 40 mg Eq gallic acid / g of sample.
 10
[13]
13. Mango seed extract according to claim 11-12 having a flavonoid content between 2 and 3.5 mg Eq catechin / g sample.

[14]
14. Use of the extract according to any of claims 11-13 as an antioxidant.
 fifteen
[15]
15. Food, cosmetic or pharmaceutical product comprising the extract of claims 11-13.

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ES2580761B1|2017-07-07|

引用文献:

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AU2019236633A1|2019-08-28|2021-03-18|Seowon University Institute of Industry - Academy Collaboration|Cosmetic composition comprising Mangifera indica irwin seed extract|

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ES201530241A|ES2580761B1|2015-02-25|2015-02-25|Procedure for obtaining an extract of mango seeds, extract thus obtained and uses given to it|ES201530241A| ES2580761B1|2015-02-25|2015-02-25|Procedure for obtaining an extract of mango seeds, extract thus obtained and uses given to it|