JP2000073056A - Oxidation resistant extract and production thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject extract capable of exhibiting an excellent oxidation resistance which is equivalent to or more than that of a red wine, and having a preventive effect to a disease in a good efficiency by extracting strained lees produced in a wine production process, after fermenting it in advance. SOLUTION: This oxidation resistant extract is obtained by fermenting strained lees produced by a production process of a wine (preferably a rose wine) at preferably 10-35 deg.C for 1-14 days and then extracting by using preferably water. Further, the fermentation can be done by utilizing an yeast contained in the strained lees. The extract can be used as it is, but it is more preferably to use after concentrating the extract upto a desired concentration under a reduced pressure, etc., for removing contained alcohols in view of avoiding a harm to a health caused by the alcohols. The extract can be suitably used e.g. by making a desired amount thereof to be contained in a cholesterol oxidation-suppressing agent, a peroxy lipid-suppressing agent, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to an antioxidant extract and a method for producing the same. More specifically, a method for producing an antioxidant extract that can be suitably used for foods, cosmetics, and the like, an antioxidant extract obtained by the method, and a cholesterol oxidation inhibitor containing the antioxidant extract , And lipid peroxide inhibitors.

[0002]

2. Description of the Related Art In recent years, active oxygen generated in a living body oxidizes lipids and proteins in the living body, and the resulting peroxide is considered to be a cause or an aggravating factor for diseases such as heart disease and allergy. It has been reported that the production of the peroxide is suppressed by taking an antioxidant.

[0003] As the above-mentioned antioxidants, chemical substances such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) have been known. However, when these substances are taken in a large amount into the body, they cause carcinogenesis. It has been pointed out that there is a possibility of exhibiting sex.

[0004] Therefore, recently, extraction of natural antioxidants from plants such as grapes, herbs, spices, and tea has been studied. In particular, red wine made from grapes is considered to exhibit excellent antioxidant action because it contains polyphenols. For example, French people who consume large amounts of red wine made from red grape varieties are generally known to have a lower mortality rate from coronary heart disease than other people [S Renault and M.D.・ Lorgeril (S.
RENAUD and M. DE LORGERIL) "Lancet"
339, 1523-6 (1992)]. This is because polyphenols, which are antioxidants derived from grapes, contained in red wine are low-density lipoproteins (LDL) in blood.
It is thought to be due to its excellent effect of inhibiting cholesterol oxidation (EN FRANKEL et.al., Lancet (Lance
t) '' Volume 341 pp. 454-7 (1993), Kay Kondou et al. (K.
KONDO et. Al.), Lancet, 344, 1152
(1994)].

[0005] However, since red wine contains about 9 to 12% of alcohol, when a Japanese person having a low alcohol metabolism ability, compared to Europeans and Americans, consumes a large amount of red wine, there is a possibility that health may be adversely affected. . Therefore,
Extraction of only antioxidants from red wine has been considered, but red wine contains a large amount of water, and thus requires a complicated operation to extract only the antioxidants. However, there is a major drawback in that a large cost is required.

In order to eliminate such drawbacks, methods for obtaining an antioxidant substance from grapes, instead of extracting from red wine, have been studied. For example, grape juice, wine pulp peel and seeds, etc. A method of extracting an antioxidant substance containing polyphenols as a main component using water or ethanol has been proposed (JP-A-7-228868, JP-A-3-200781).

[0007] However, the antioxidants extracted by the method have a drawback that they do not exhibit sufficient antioxidant properties.

[0008] In recent years, a method for extracting polyphenols from grape seeds has been proposed, but it is known that the extract obtained by such a method is not the same as the antioxidant substance of red wine. .

Therefore, recently, development of a method capable of efficiently producing an extract containing a large amount of an antioxidant having excellent antioxidant properties, such as red wine, has been desired.

[0010]

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above-mentioned prior art. From the viewpoint of enhancing the preventive effect against diseases, an antioxidant substance having an antioxidant property equal to or higher than that of red wine is provided. An object of the present invention is to provide an antioxidant extract containing a large amount and a method capable of efficiently producing the antioxidant extract.

[0011]

The gist of the present invention is [1].
After fermenting the compressed juice lees generated in the wine manufacturing process, a method for producing an antioxidant extract, which is characterized by being extracted,
And [2] an antioxidant extract obtained by the above-mentioned production method.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, compressed squeezed cake produced at the stage of producing wine is used.

In the present invention, there is one major feature in using this compressed juice cake.

Heretofore, compressed squeezed cake has been used as a part for extracting pigments and alcohols and as feed for livestock, but has been generally discarded as unnecessary. The present invention uses the compressed squeezed cake as a starting material. The compressed squeezed cake is generally inexpensive and easily available, and therefore is a very economical method.

The present invention also has one major feature in that the above-mentioned compressed juice cake is fermented in advance.

[0016] Generally, in the compressed squeezed cake produced during the production of wine, most of the antioxidant extract as an active ingredient is transferred to wine, so that the same antioxidant extract as red wine is used. It is unlikely that it is contained in large quantities.
Therefore, even if the compressed squeezed cake is extracted with an extraction solvent such as water or alcohol as in the related art, it is considered that the resulting extract does not contain much antioxidant extract similar to red wine. Can be

However, when an operation of fermenting the compressed juice cake in advance instead of immediately extracting it with the extraction solvent is employed, the compressed juice cake is mixed with the antioxidant substance contained in the red wine. It is possible to produce very efficiently an extract containing a large amount of antioxidant substances exhibiting equal or superior antioxidant properties. It is.

Furthermore, among the compressed squeezed lees, particularly when the compressed squeezed lees of rose wine is used, a large amount of the antioxidant substance is contained as compared with the case where the compressed squeezed lees of red wine is used. An antioxidant extract can be obtained, and the antioxidant contained in the antioxidant extract has an antioxidant that is at least as good as the antioxidant contained in red wine. Has several excellent effects.

The method for producing wine used in obtaining the compressed squeezed cake used in the present invention is not particularly limited as long as it is a generally employed method. As a wine production method,
An outline of one embodiment of a typical method for producing rose wine will be briefly described below.

A red grape variety is used as a raw grape for rose wine.

Specific examples of the red grape varieties include, for example, cabernet franc, cabernet sauvignon,
Merlot, Pinot Noir, Pinot Meunier, Muscat Berry A, Syrah, Gamay, Grenache, Mourvèdre, Cinsault, Glowau and the like. These can be used alone or in combination of two or more.

After the raw grapes are crushed and destemmed, metabiose potassium and yeast are added thereto. The obtained mixture is fermented at a temperature of about 25 ° C. for a short period of about one day or less, and then squeezed to remove pulp, seeds and skin (compressed juice cake). Perform fermentation around days. Next, transfer the obtained fermented product to a keg or storage tank, ripen it for a short time while pulling it,
By lowering, then filtering and bottling, a bottled wine can be obtained.

The most desirable antioxidant extract of the present invention can be obtained by first fermenting and extracting the compressed juice cake produced in the process of producing rose wine using the above-mentioned red grape seeds.

The fermentation of the compressed juice lees can be carried out using yeast contained in the compressed juice lees obtained in the wine manufacturing process. As a representative example of the yeast, for example,
Saccharomyces cerevi
siae), Saccharomyces cerevisiae var. elli.
psoideus). Yeast is usually contained in the compressed juice lees in an amount of 1.0 × 10 ³ to 1.0.g per g of the compressed juice lees.
It is preferable to contain about 0 × 10 ⁹ strains.

The fermentation temperature of the compressed juice lees is usually 10 to 35 ° C., preferably 2 to 35 ° C., because the yeast is mesophilic.
Desirably, the temperature is 0 to 30 ° C. In addition, the fermentation period of the compressed squeeze cake cannot be determined unequivocally because it varies depending on the fermentation temperature and the like, but from the viewpoint of performing fermentation sufficiently, it is preferably at least 1 day, preferably at least 3 days, In addition, from the viewpoint of preventing deterioration of the raw material due to propagation of other bacteria and mold, the period is 14 days or less, preferably 10 days or less.
It is desirable to keep it for no more than days.

After completion of the fermentation, the compressed juice cake can be used as it is or after being crushed to a desired size without breaking the seeds as necessary. In addition, when crushing the compressed juice lees, for example, a cutting mill or the like can be used.

When performing the extraction, any of a batch-type extraction device and a continuous-type extraction device can be used as the extraction device.

In the present invention, water can be suitably used as the extraction solvent. The type of such water is not particularly limited, and pure water, purified water, and the like can be used.
The amount of the extraction solvent used is usually about 100 to 1000 parts by weight, preferably 30 to 100 parts by weight of the compressed juice cake.
It is desirable that the amount is about 0 to 600 parts by weight.

Extraction is performed after mixing the compressed squeezed cake and the extraction solvent. The extraction temperature at this time is from 20 to 80 ° C., preferably from 40 to 60 ° C., from the viewpoint of efficient extraction.
It is desirable that Further, the extraction time cannot be determined unequivocally because it differs depending on the extraction temperature or the like, but is usually about 2 to 24 hours.

After completion of the extraction, the extract may be separated into solid and liquid by means such as filtration, centrifugation, or filter press.

Thus, the antioxidant extract of the present invention is obtained. The antioxidant extract contains polyphenols, sugars, some alcohols, and the like.

The antioxidant extract of the present invention can be used as it is, but may be appropriately concentrated to a desired concentration by, for example, concentration under reduced pressure. When such concentration is performed, alcohol contained in a small amount, though not as much as red wine, can be removed simultaneously with concentration. Therefore, in the present invention, it is preferable to concentrate the antioxidant extract from the viewpoint of avoiding health problems due to alcohol.

Since the antioxidant extract of the present invention is a natural extract, it can be used by adding it to food.
It can be suitably used for cosmetics and the like. Further, the antioxidant extract of the present invention is, for example, by mixing with an excipient, freeze-drying, by drying by spray drying, etc.,
It can be prepared in a powder state and used for various purposes.

Since the antioxidant extract of the present invention contains an antioxidant substance represented by polyphenol similarly to red wine, the antioxidant effect of low-density lipoprotein cholesterol and the antioxidant ability of serum are increased. It exerts an excellent action such as an action of suppressing fatty acid conversion in blood. Therefore,
For example, by using the antioxidant extract of the present invention,
When producing the same effect as red wine, it is generally considered that French citizens are said to have lower coronary heart disease mortality than nationals in other countries based on their constant consumption of red wine. Thus, the French people's wine intake can be taken into account. The average daily red wine consumption of French citizens is generally 200-400 ml.
It is said that it is about. From this, when expecting the same effect as red wine, it is necessary to ingest the antioxidant extract in an amount corresponding to the antioxidant contained in the average red wine intake of 200 to 400 ml per day. It is considered preferable.

The antioxidant extract of the present invention can be suitably used by adding a desired amount thereof to, for example, a cholesterol oxidation inhibitor, a lipid peroxide inhibitor and the like.

[0036]

Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to only these examples.

Example 1 Compressed squeezed cake 100 k produced during production of rose wine
g (raw grape variety: Grenache) at 15-25 ° C
After fermentation for 7 days at room temperature, this was mixed with 600 kg of water and then subjected to an extraction treatment for 10 hours.

The obtained extract was subjected to solid-liquid separation with a filter press, and concentrated under reduced pressure until the soluble solid content was reduced to 55% by weight while removing alcohol to obtain Extract A. The resulting extract A was a reddish-brown, slightly turbid, fluid liquid with a pH of 3.5, a tartaric acidity of 8% by weight, and a sourness and astringency.

Next, the polyphenol content of the extract A was measured by the Folin-DENIS method using gallic acid as a standard substance. The polyphenol content was 800 mg / 100 g. Aldrin, dieldrin, endrin, parathion, DDT, BHC and mold venom aflatoxin B1 are used as pesticide residues.
Was not detected from the extract A. In addition, arsenic, cadmium, lead, tin and coliforms contained in the extract A all conform to the component standards of the Ministry of Health and Welfare Notification No. 370, “Food and Additive Standards First Food D Each Article Soft Drinking Water”. It was compatible.

Experimental Example 1 [Antioxidant Property of Extract] The antioxidant property of extract A obtained in Example 1, that is, the superoxide scavenging activity (hereinafter, referred to as SOSA) was evaluated as ES.
It was examined by the R method.

On the other hand, as a control of extract A, extract B was prepared in the same manner as above except that no fermentation was carried out when producing the antioxidant extract. After adjusting the polyphenol content contained in the obtained extract B by diluting with water so as to be equal to the extract A,
The SOSA was examined.

The ESR method is an abbreviation of electron spin resonance (Electron Spin Resonance). A sample having unpaired electrons is placed in a magnetic field and irradiated with microwaves to absorb electromagnetic waves due to resonance. It is a method of measuring.

[0043] SOSA can be easily measured by the ESR method by using a spin trap agent for trapping superoxide. Conventional cytochrome C method or N
The BT reduction method has a drawback that when using an unpurified sample solution, the effects of peroxidase and coexisting reducing substances cannot be neglected. Recently, this ESR method is often used for measuring antioxidant properties. Used [Japan Food Research Laboratories "JFRL News" 50 volumes (199
February 3)].

As a result of examining the SOSA, the SOSA of the extract B extracted without fermentation was 4.8 × 10 ³ units / g, whereas the SOSA of the extract A extracted after fermentation was 4.8 × 10 ³ units / g.
Was 9.4 × 10 ³ units / g.

This indicates that the fermentation of the compressed juice lees enhances the antioxidant properties of the extract.

Experimental Example 2 [Oxidation Inhibiting Effect of LDL] When LDL, which is a low-density lipoprotein, undergoes oxidative denaturation, it is not processed in the usual metabolic process via the LDL receptor, and is taken up by macrophages. It is known that macrophages that have taken up a large amount of oxidized LDL become foam cells and accumulate in the blood vessel wall to cause arteriosclerosis [see Toshiichi Yoshikawa, "Science of Free Radicals", Kodansha (1997). ].

It is known that the LDL solution is rapidly oxidized in the presence of copper ions or iron ions.
Therefore, when an antioxidant is added to the LDL solution, the oxidation of LDL is suppressed for a longer time as the antioxidant of the antioxidant is stronger, so that the time when the oxidation of LDL starts is delayed.

Therefore, the degree of oxidation of LDL can be evaluated by the amount of malondialdehyde, which is an oxidized metabolite of lipid contained in LDL. Therefore, the extract A obtained in Example 1 As a control, the antioxidant properties of red wine were compared using a human LDL oxidation induction model. Specifically, first, LDL was separated by ultracentrifuging human serum at a temperature of 15 ° C. at 650000 g × 2 hours. The ultracentrifuge used was a product made by Beckman, trade name: L90 (rotor: NVT 90 Titanium rotor). After that, the final concentration of LDL obtained was 50 μg protein / m
Purified water was added to this so as to obtain 1.

Next, extract A and red wine (Cabernet-INRA) were each diluted with purified water containing 12% alcohol to obtain a dilute solution of extract A and a dilute solution of red wine. Each of the obtained diluents was mixed with the LDL solution to obtain a mixture having a polyphenol content shown in Table 1.

Next, each mixture was incubated at 37 ° C. for 30 minutes, and then copper chloride was added to each mixture so that the content of copper chloride (CuCl ₂ ) was 5 μM in order to promote the oxidation of LDL. After the addition, keep at 30 ° C.
The time until the oxidation of the compound started was measured. In addition, for the time until the oxidation of LDL starts, after adding copper chloride to each mixed solution, the amount of conjugated diene, which is an intermediate product of lipid oxidation, is measured using an absorbance meter [Kontron, trade name: Uvi
kon 820], and the absorbance at 234 nm was measured over time, and the time required until the absorbance of each mixed solution started to increase was defined as the time required. Table 1 shows the results.

[0051]

[Table 1]

From the results shown in Table 1, the extract A and the red wine both delay the onset of LDL oxidation in proportion to the polyphenol concentration. It can be seen that it has. Further, the extract A is compared with red wine, LDL
Since the oxidation onset time is equal to or longer than
It can be seen that the red wine has the same or better antioxidant properties as red wine, which is said to be excellent in antioxidant properties.

Experimental Example 3 [In vivo antioxidant properties] The antioxidant properties when the extract A was ingested into a living body were examined as follows.

First, 10 rats were prepared for each test group, drinking water and food were prepared as feeds to be given to the rats, and these rats were allowed to freely ingest these at the same time.

In order to confirm the effect of the extract A, the extract A was not added at all to the drinking water and the diet (control), and the extract A was added only to the drinking water ( (Polyphenol content: 3.26 g / l)
[Administration group I], and when extract A was added only to the diet (polyphenol content: 4.30 g / kg) [Administration group
II], an experiment was conducted as follows.

After feeding the rats with the above-mentioned feed for 12 weeks and raising them, serum was collected. The total antioxidant activity of the serum was measured in order to confirm that the components of the extract A were absorbed and entered the blood to improve the antioxidant properties of the blood. Total antioxidant activity of serum is ABT which is the same radical as active enzyme
It was evaluated by the activity of scavenging the S [2,2'-azinobis (3-ethylbenzothiazoline-6-sulfate)] radical cation.

As a result, the total antioxidant activity of the serum was 0.78 ± 0.03 mmol / ml in administration group I and 0.79 ± 0.03 mmol / ml in administration group II,
In the control, it was 0.60 ± 0.02 mmol / ml.

From the above results, the total antioxidant activity of the serum is increased regardless of whether the extract A is added to drinking water or diet, so that the extract A is administered orally to It can be seen that the oxidizing property can be improved.

EXPERIMENTAL EXAMPLE 4 [Lipid Oxidation Inhibiting Effect of Blood] In order to confirm the effect of the extract A on inhibiting lipid oxidation of blood, the extract was added to both drinking water and diet as a feed in the same manner as in Experimental Example 3. A to which no extract A was added [control] and those to which extract A was added only to drinking water (polyphenol content: 3.26 g / l) [Administration group III
] Was prepared, and an experiment was performed as follows.

[0060] Ten rats were prepared for each test group, and each rat was allowed to freely ingest the feed for 12 weeks and bred, and then serum was collected. In order to examine whether the components of the extract A exhibited the effect of suppressing lipid oxidation in blood, the amount of malondialdehyde, which is an oxidation metabolite of lipids contained in blood, was measured. The content of malondialdehyde in serum was as follows: malondialdehyde and N-methyl-2-
It was determined by measuring the absorbance at a wavelength of 586 nm of a stable compound obtained by reacting with phenylindole.

As a result, the content of malondialdehyde in the serum was 1.27 ± 0.03 μmol / m in the administration group III.
l, whereas the control is 1.60 ± 0.13 μmo
1 / ml.

From the above results, when extract A was added to the diet, the content of malondialdehyde in the serum was reduced, so that extract A exhibited an effect of suppressing blood lipid oxidation by oral administration. It turns out that it is.

[0063]

According to the production method of the present invention, it is possible to efficiently produce an antioxidant extract containing a large amount of an antioxidant having excellent antioxidant properties.

The antioxidant extract of the present invention is a natural extract and contains a large amount of an antioxidant having excellent antioxidant properties, so that it can be widely used for various uses such as foods and cosmetics. What can be used.

 ──────────────────────────────────────────────────の Continuing on the front page (72) Inventor Majour André France 63800 Cournon, Rue des Perbanches 10 (72) Inventor Lock Edmon France 63170 Aubiere, Shman de la Ganne, Amour les Aigrantier 9F Term (reference) 4C088 AB56 AC14 BA08 BA09 CA02 CA25 NA05 ZC21 ZC33 4H025 BA01

Claims (8)

[Claims] 1. A method for producing an antioxidant extract, comprising fermenting and extracting compressed juice cake produced in a wine production process. 2. The method for producing an antioxidant extract according to claim 1, wherein the wine is rose wine. 3. The method for producing an antioxidant extract according to claim 1, wherein the compressed squeezed cake is fermented at a fermentation temperature of 10 to 35 ° C. 4. The method for producing an antioxidant extract according to claim 1, wherein the compressed juice cake is fermented for 1 to 14 days. 5. The method for producing an antioxidant extract according to claim 1, wherein water is used as an extraction solvent during the extraction. 6. An antioxidant extract obtained by the method according to any one of claims 1 to 5. 7. An LDL cholesterol oxidation inhibitor comprising the antioxidant extract according to claim 6. 8. A lipid peroxide inhibitor comprising the antioxidant extract according to claim 6.