KR20210099897A - Manufacturing method of grape fermented composition containing a large amount of polyphenols - Google Patents

Abstract

The present invention relates to a method for preparing a grape fermented product containing plenty of polyphenols. Provided is a grape fermented product containing plenty of polyphenols by using bunch stems of grapes which are byproducts of the grapes. Also, the present invention can provide a grape fermented product which contains the maximum amount of polyphenols depending on grape juice, adding amounts of bunch stems, and fermentation conditions.

Description

The manufacturing method of grape fermented composition containing a large amount of polyphenols {Manufacturing method of grape fermented composition containing a large amount of polyphenols}

The present invention relates to a method for producing a fermented grape product containing a large amount of polyphenols.

Medically, antioxidant refers to the process of preventing damage to cells in our body due to oxidation. Active oxygen is a substance produced in the process of human respiration and is essential for maintaining life, but it can also cause diseases by damaging cells such as skin or blood vessels. Vitamin C, vitamin E, and phytochemicals contained in plants are known as representative antioxidants.

Polyphenols are a kind of phytochemical, a kind of defense substance that plants create to protect themselves from UV rays. Although French people eat a lot of meat, cardiovascular disease is one-third that of Americans, because it is known that polyphenols in wine, which French people drink frequently, protect human cells. Polyphenols are abundantly contained in grapes, wine, and blueberries, and representative substances include catechin, anthocyanin, resveratrol, quercetin, and the like.

Grapes contain polyphenols such as anthocyanins and resveratrol. Anthosingin prevents the formation of blood clots, improves blood circulation, and prevents heart disease. In addition, research results are also known to prevent vascular dementia because it improves blood flow. On the other hand, resveratrol has excellent effects in preventing skin aging and relieving inflammation in the body such as the scalp, and has the effect of protecting the cardiovascular system by increasing the activity of catalase, an enzyme substance that suppresses and prevents stress.

Republic of Korea Patent Registration No. 10-0893788 (2009.04.09.) describes a method for purifying resveratrol with high purity from the extract of grape clusters. Republic of Korea Patent No. 10-1273426 (2013.06.04.) describes a method for producing a feed additive for livestock or fish, in which polyphenols are fortified using grape by-products and propolis extract.

An object of the present invention is to provide a grape fermented product containing a large amount of polyphenols by utilizing cluster branches of grapes, which are by-products of grapes.

In addition, an object of the present invention is to provide an efficient method for producing a fermented grape product by comparing the polyphenol content according to the method for producing a fermented grape product.

The present invention comprises the steps of: (a) preparing grape juice by squeezing grape kernels, and adding 1 to 10% by weight of cluster eggplant to the grape juice with respect to the grape juice; and (b) fermenting for 4-10 days at 25-30° C. after step (a).

In the present invention, the method for producing the fermented grape product, preferably, after the step (b), the step (c) of aging at 2 to 8 ℃ for 10 to 60 days; it is good to further include.

In the present invention, the grape juice of step (a) is preferably juiced, and then sugar is added to adjust the sugar content to 20 to 25 Brix.

In the present invention, the cluster branches of step (a) are preferably completely immersed so as not to float to the top of the grape juice.

The present invention provides a grape fermented product containing a large amount of polyphenols by utilizing cluster branches of grapes, which are by-products of grapes. Pine branches contain a large amount of polyphenols, so a grape fermented product containing a large amount of polyphenols can be prepared by using the pine branches.

In addition, the present invention can provide a grape fermented product containing the maximum amount of polyphenols according to the grape juice, the addition amount of pine needles, and the fermentation conditions.

1 is a graph showing the resveratrol content according to sweetened fermentation of grape juice.
2 is a graph showing the content of resveratrol and alcohol according to the method for preparing grape juice.
3 is a graph showing the content of resveratrol according to the addition amount of pine needles.
4 is a graph showing the content of resveratrol by grape variety by fermenting the grape fermented product at 25°C. (A) is Meruru grape (MBA), (B) is Seridan grape.
5 is a graph showing the content of resveratrol by grape variety by fermenting the grape fermented product at 30°C. (A) is Meruru grape (MBA), (B) is Seridan grape.
6 is a graph showing the content of resveratrol in the case of preparing a fermented grape product using Meruru grapes (MBA).
7 is a graph showing the content of resveratrol in the case of mass production of fermented grapes using Meruru grapes (MBA).
8 is a graph showing the results of measuring the resveratrol content at 10-day intervals by aging the grape fermented product for 60 days.
9 is a standard curve of gallic acid as a standard material.
10 is a graph showing the change in absorbance and antioxidant effect according to the reaction time of the grape fermented product and the DPPH solution.

An object of the present invention is to provide a fermented grape product containing a large amount of polyphenols. Specifically, the present invention comprises the steps of: (a) preparing a grape juice by squeezing grape grains, and adding 1 to 10% by weight of cluster eggplant to the grape juice with respect to the grape juice; and (b) fermenting for 4-10 days at 25-30° C. after step (a).

Grape clusters contain polyphenols such as resveratrol. As a result of resveratrol eluting by drying and pulverizing cluster branches in the present invention, it was analyzed that 299 μg/g of Campbell Early grapes and 276 μg/g of resveratrol were contained in cluster branches of Seridan grapes.

In the present invention, the grape juice of step (a) is preferably juiced, and then sugar is added to adjust the sugar content to 20 to 25 Brix. In general, when grape kernels are crushed and squeezed with a grinder, about 13 Brix of grape juice can be obtained. In the present invention, it was confirmed that the content of resveratrol was increased when sugar was added to grape juice for sweetened fermentation.

In the present invention, the grape juice is preferably obtained by pulverizing and juicing with a grinder. In the present invention, it was confirmed that the content of resveratrol was low when grape juice was obtained by crushing grape kernels by hand.

In the present invention, it is preferable to add 1 to 10% by weight of cluster eggplant with respect to the grape juice, preferably 5% by weight of cluster eggplant with respect to the grape juice. As shown in FIG. 3 of the present invention, it was confirmed that the content of resveratrol increased in proportion to the fermentation time when 5% by weight of cherry eggplant was added.

In the present invention, it is preferable to perform fermentation by inoculating yeast during the fermentation.

On the other hand, when carbon dioxide and alcohol start to be produced after the start of fermentation, the relatively large grains of grape skins and crushed branches rise to the top and are separated from the grape juice, and it was found that it is difficult to further elution of resveratrol, that is, polyphenols. Accordingly, the present invention was to maximize the dissolution of polyphenols by increasing the area or time during which the cherry blossom branches can come into contact with the grape juice during fermentation. Therefore, preferably, in order to prevent the cluster branches from floating to the top of the grape juice, it is preferable to wrap the crushed cluster branches in gauze together with a pestle (porcelain) and completely immerse them in the grape juice for fermentation.

In the present invention, the grape fermented product is preferably fermented at 25 ~ 30 ℃, more preferably it is good to ferment at 30 ℃. When fermented at 25°C, the resveratrol content of Meruru grapes (MBA) is significantly higher, but it is preferable to ferment at 30°C in consideration of the alcohol generation rate.

In the present invention, the grape fermented product is preferably fermented for 4 to 10 days. In particular, it was confirmed that the content of resveratrol showed the maximum value on the 8th day of fermentation when 5% by weight of pine tree eggplant was added.

In the present invention, the method for producing the fermented grape product, preferably, after the step (b), the step (c) of aging at 2 to 8 ℃ for 10 to 60 days; it is good to further include. When the grape fermented product of the present invention was aged for 10 days, the content of resveratrol was maximum, and it was confirmed that resveratrol was stably present until the 60th day of aging.

On the other hand, the fermented grape of the present invention was found to have a significantly higher polyphenol content than that of ordinary wine, and thus the antioxidant effect was also excellent. In particular, even when the grape fermented product of the present invention was diluted to 1/50 level, it showed a high antioxidant effect of 52.4% compared to the positive control.

Hereinafter, the contents of the present invention will be described in more detail through the following Examples and Experimental Examples. However, the scope of the present invention is not limited only to the following examples and experimental examples, and includes modifications of technical ideas equivalent thereto.

[Experimental Example 1: Analysis of the content of resveratrol contained in pine needles]

After drying and pulverizing cluster branches of Campbell Early and Sheridan grapes at 60°C for 24 hours, 1 g of this is taken and 10 ml of 20% (v/v), 30% (v/v), Each was mixed with 40% (v/v) ethanol. Resveratrol was eluted from the sample using an ultrasonic crusher. After mixing the same amount (10ml) of diethyl ether, it was vortexed for 1 minute in a separatory funnel, and the process of recovering the supernatant (ether layer) was repeated 5 times. After evaporating all of the ether solvent at 40° C. using a vacuum rotary concentrator to the last recovered supernatant, 1 ml of acetonile was added and centrifuged at 14,000 rpm for 20 minutes. The recovered supernatant was subjected to high-performance liquid chromatography (HPLC). ) was analyzed for resveratrol content.

Run Time Flow Rats
(ml/min) Solvent ration Acetonitrile (%) Water (%) 0 0.3 30 70 10 0.3 30 70 20 0.3 50 50 45 0.3 50 50 50 0.3 30 70 55 0.3 30 70

As a result of the analysis, it was analyzed that the cluster branches of Campbell Early grapes contained 299 μg/g of resveratrol, and the cluster branches of Seridan grapes contained 276 μg/g of resveratrol. Through this experiment, it was confirmed that a large amount of resveratrol was also contained in grape cluster branches.

[Example 1: Comparison of resveratrol content according to sweetened fermentation of grape juice]

Campbell early grape kernels were crushed and squeezed with a grinder to obtain about 13Brix of grape juice. The contents of resveratrol were compared by inoculating yeast and fermenting for 8 days at 25°C after adding grape juice to two 10L fermenters and adding sugar to the final concentrations of 20Brix and 25Brix, respectively.

As shown in Figure 1, the resveratrol content was the highest on the 6th day of fermentation, and the content of resveratrol was higher in 25Brix than in 20Brix. confirmed that there is. Through this, it was determined that adding sugar to the grape juice was a suitable technique for enhancing the resveratrol content.

[Example 2: Comparison of resveratrol content according to the presence or absence of grape branch and juicing method]

With respect to Campbell's Early grape juice, 5 wt % of clustered eggplant was added, and then pulverized and squeezed with a grinder to obtain grape juice to which clustered branch was added (▼ in FIG. 2 ). In addition, only grape grains were crushed and juiced with a grinder (homogenizer) to obtain grape juice (circle in FIG. 2 ). As a control, grape juice obtained by crushing grape kernels by hand was used ( ● in FIG. 2 ). Each grape juice was placed in a 10L fermenter, inoculated with yeast, and fermented at 25°C for 16 days to quantify the resveratrol content.

As shown in FIG. 2, all three methods showed no significant difference in the trend of increasing alcohol content and the maximum alcohol content (FIG. 2B). Therefore, it could be evaluated that the difference in the extraction method (using homogenizer vs. crushing by hand) did not show any significant results.

On the other hand, the content of resveratrol increased from the 2nd day of fermentation in the grape juice to which pine branches were added, and the resveratrol content was maintained even after the 8th day of fermentation. showed shape. Therefore, it could be evaluated that the addition of cluster eggplant exhibits an excellent effect in increasing the resveratrol content.

[Example 3: Comparison of resveratrol content according to the amount of pine eggplant added]

To the Campbell Early grape juice, 1, 5, and 10% by weight of pine branches were added, respectively, and then pulverized and juiced with a grinder to obtain a grape juice with pine branches added. The obtained grape juice was placed in a 10L fermenter, inoculated with yeast, and fermented at 25° C. for 8 days to quantify the content of resveratrol.

As shown in FIG. 3 (1 wt% of cluster eggplant: , 5 wt% of cluster eggplant: ○, 10 wt% of cluster eggplant: ▼), there was no significant increase in resveratrol according to the amount of cluster eggplant added. It was determined that this is because when carbon dioxide and alcohol start to be produced after the start of fermentation, the large grains of grape skins and crushed cluster branches float to the top and are separated from the grape juice, and no further elution occurs.

Therefore, rather than increasing the amount of pine branches added, it was attempted to find a way to increase the area or time during which pine branches can come into contact with the grape juice during fermentation.

[Example 4: Comparison of resveratrol content according to grape varieties and fermentation conditions]

To compare the resveratrol content according to grape varieties (MBA) and Seridan grapes, 1% by weight of each grape cluster was added to the MBA and Seridan grape juices, and the content of resveratrol was measured by inoculating yeast. The fermentation temperature was changed to 25 °C and 30 °C. The case fermented at 25°C is shown in FIG. 4 and the case fermented at 30°C is shown in FIG. 5 .

As shown in Figures 4 and 5, when the resveratrol content is fermented at 30 ℃, the resveratrol of the MBA (Fig. 5 (A)) variety is up to 3.86 mg/L, and the Seridan (Fig. 5 (B)) variety is about As it contained 3 mg/L of resveratrol, it was confirmed that the resveratrol content was superior to that of Campbell grapes fermented at 25°C. In addition, when fermented at 30°C, the rate of alcohol generation was faster than that of fermentation at 25°C.

From the above experiment, it was confirmed that the content of resveratrol was higher in Meruru grapes and Seridan grapes than the Campbell grapes, and it was confirmed that the fermentation condition at 30° C. was more suitable.

[Example 5: Comparison of resveratrol content according to immersion of crushed pine branches]

From Example 3 above, it was attempted to increase the content of resveratrol by increasing the area or time during which the pine cones could come into contact with the grape juice during fermentation. Accordingly, in this implementation, a method was sought to prevent the pine cones from rising to the top during fermentation.

1 wt%, 2 wt%, and 5 wt% of mulberry grapes (MBA) were added to 10 kg of the grape marsupial (MBA) juice, respectively, and yeast was inoculated and fermented at 30° C. for 10 days. In order to prevent the pine branches from floating to the top during fermentation, the crushed pine branches were wrapped in gauze together with a pestle (porcelain) and immersed in the grape juice.

As shown in FIG. 6 , when 5 wt% of pine eggplant was added on the 8th day of fermentation, the content of resveratrol showed a maximum of 5.32 mg/L, and when 1 wt% of pine eggplant was added, 5.03 mg/L, 2 wt% When added, it was found to be 4.97 mg/L. This is an increase of more than 1.3 times compared to the maximum content of 3.86 mg/L in the case of not immersing the cluster branches (see Example 4). However, as shown in Example 3, considering that there is no significant increase in resveratrol depending on the amount of the eggplant added, it is desirable to add 1% by weight of the grape juice to the cluster eggplant for the efficiency of the manufacturing process. became

[Example 6: Mass fermentation of grape juice]

The resveratrol content was measured by mixing 80 kg of grape marsupial (MBA) juice and 0.8 kg of crushed mulberry branch of maru grapes (MBA), inoculating yeast, fermenting it for 8 days in a constant temperature room at 30° C., and filtering it. In order to prevent the pine branches from floating to the top during fermentation, the crushed pine branches were wrapped in gauze together with a pestle (porcelain) and immersed in the grape juice.

As shown in FIG. 7 , it was found that resveratrol was contained at a maximum of 4.39 mg/L on the 8th day of fermentation.

[Experimental Example 2: Change in resveratrol content during aging period]

The fermented grape obtained in Example 6 was moved to a low temperature room (4° C.), sealed so that air does not pass through, and aged for 60 days, and the resveratrol content was quantified at 10-day intervals.

As shown in FIG. 8 , the content of resveratrol showed a tendency to decrease slightly according to the aging period, but a rapid change was not observed. In particular, it was confirmed that 3.24 mg/L of resveratrol remained even after 60 days, and thus, it was confirmed that resveratrol can be stably present even when the present fermented product is distributed for a long time.

[Experimental Example 3: Confirmation of the functionality of fermented products containing high resveratrol]

In this experiment, the polyphenol content was compared to compare the functionality of the grape fermented product obtained in Experimental Example 2 and aged at low temperature for 60 days and the normal wine.

Polyphenol quantification was used by modifying the Folin-Denis method. That is, 6.5 ml of distilled water was added to 0.5 ml of the sample, 0.5 ml of Folin-ciocalteu's phenol reagent (Sigma) was added, and left for 3 minutes. After adding 1ml of sodium carbonate and developing color for 30 minutes, absorbance was measured at 725mn to quantify polyphenol content. As a standard material, gallic acid, which is abundantly contained in grapes, was used, and the standard curve is shown in FIG. 9 . Fermented grapes aged for 60 days of the present invention as regular wine (sample), Beaujolais (France, 2004), Cabernet souvignon (USA, 2003), Point noir (Chile, 2003), Noel wine (Korea, 2003), Chateau Mani (domestic production, 2004) was purchased from the market and used.

The standard material, gallic acid, showed a linearity up to 0.25 mg/ml, and the correlation (r) was 0.9985. The results of quantifying the polyphenol content of each sample are shown in Table 2 below.

sample Polyphenol content (mg/L) Fermented grapes aged for 60 days of the present invention 3.49 Beaujolais (France, 2004) 2.66 Cabernet souvignon (USA, 2003) 2.85 Point noir (Chile, 2003) 1.10 Sunset Wine (Korea, 2003) 1.86 Chateau Mani (Korea, 2004) 2.19

As a result of quantifying the polyphenol content of each sample, it was found that the fermented grapes aged for 60 days of the present invention contained significantly more polyphenols than that of general wine. This was judged to be the result of dissolution of a large amount of polyphenols contained in pine branches.

[Experimental Example 4: Confirmation of antioxidant effect of fermented products with high resveratrol content]

In this experiment, it was attempted to confirm the antioxidant effect of the grape fermented product obtained in Experimental Example 2 and aged at low temperature for 60 days.

First, the sample and the DPPH solution were mixed (check the mixing ratio) to determine the appropriate reaction time required for scavenging DPPH free radicals in the grape fermented product, and then the absorbance was measured at 5 minute intervals up to 30 minutes to observe the change over time. .

As shown in Table 3 and FIG. 10, the absorbance showed the lowest value after 10 minutes of mixing the sample and the DPPH solution, and showed a tendency to increase thereafter, so the optimal reaction time was determined to be 10 minutes.

time (minutes) Absorbance (517nm) 0 (before sample addition) 0.8177 5 0.1564 10 0.1285 15 0.1503 20 0.1656 25 0.1997 30 0.2091

To compare the antioxidant effect, gallic acid and caechin solutions, which are known to have excellent antioxidant effects, were prepared at 0.05mM, 0.1mM, 0.25mM, 0.5mM, 1mM, 5mM, and 10mM, respectively, and the antioxidant activity was verified. did. As shown in Table 6, at a concentration of 1 mM or more, it was observed that the antioxidant activity no longer increased, and it was determined that the antioxidant activity reached saturation (saturation).

Standard
matter Concentration (mM) 0.05 0.1 0.25 0.5 One 5 10 gallic acid 64.7 77.2 82.3 85.4 86.9 87.0 87.0 catechins 62.9 73.9 79.2 82.7 84.6 84.8 84.4

In order to standardize the sample, 100 ml of 7 samples were concentrated at 40° C. for 20 minutes with a vacuum distillation device to volatilize ethanol, 15 ml of 99.9% ethanol was added, and the mixture was refined to 100 ml with distilled water, and the pH was adjusted to 6.8 using 1N NaOH. did. The antioxidant effect was compared by measuring the electron donating ability (free radical scavenging effect) of DPPH. That is, 4.5 ml of DPPH solution was added to 0.5 ml of sample, and after reacting for 20 minutes, the antioxidant effect was calculated from the value (S) measured for absorbance at 517 nm and the value measured using 15% ethanol instead of the sample (C).

[Equation 1]

Antioxidative effect (%) = { 1 - (S /C} × 100

Relative Antioxidative Activity (RAA, %) sample
undiluted sample
1/10 dilution sample
1/50 dilution Gallic acid 10 mM 100 100 100 Fermented grapes aged for 60 days of the present invention 95.2 77.2 52.4 Beaujolais (France, 2004) 87.6 70.0 44.3 Cabernet souvignon
(USA, 2003) 84.1 71.9 46.5 Point noir (Chile, 2003) 73.6 49.4 20.1 Sunset Wine (Korea, 2003) 71.0 43.6 10.9 Chateau Mani (Korea, 2004) 80.3 70.8 26.6

As shown in Table 5, even when the fermented grape juice of the present invention was diluted to 1/50, the antioxidant activity was 52.4%, and it was confirmed that the antioxidant activity was significantly higher than that of ordinary wine.

Claims (4)

(a) preparing a grape juice by squeezing grape grains, and adding 1 to 10% by weight of cluster branches based on the grape juice to the grape juice; and
After the step (a), a step (b) of fermenting at 25-30 ° C. for 4-10 days;
According to claim 1,
The method for producing the grape fermented product,
After the step (b), the step (c) of aging at 2 to 8° C. for 10 to 60 days;
According to claim 1,
The grape juice of step (a) is,
A method for producing fermented grapes, characterized in that after squeezing, sugar is added to adjust the sugar content to 20-25 Brix.
According to claim 1,
The cluster branches of step (a) are,
A method for producing a fermented grape, characterized in that it is completely immersed so as not to float to the top of the grape juice.

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