KR20050103261A - Method of manufecture of the odorless functional garlic fermentation beverage (gfb) with tonic effect by natural fermentation - Google Patents

Abstract

    The present invention relates to an odorless functional garlic beverage having a tonic effect by natural fermentation and a method for preparing the same. The ratio of 1.0: 1.5 garlic and mixed solution in a mixed solution of sugar-fructose-oligosaccharide in native pottery in Korean clay pot The primary fermentation extract (A) from which garlic was extracted by primary fermentation for 25 days and garlic was dried and dried garlic for 5 days in a dryer at 30 ° C. was mixed with sugar: Into the indigenous pottery at a ratio of 1 to 2 days of natural fermentation, the second fermentation extract (B) from which the garlic was extracted, mixed with a 1: 1 ratio of garlic husks containing garlic peeled 5: Functional odorless garlic garlic fermented beverage having a tonic effect by natural fermentation, characterized in that it is prepared by mixing in a ratio of 1, and then fermented for 3 days in a plastic house for 45 days.

Description

Odorless functional garlic fermented beverage having a tonic effect by natural fermentation and its preparation method (method of manufecture of the odorless functional garlic fermentation beverage (GFB) with tonic effect by natural fermentation}

 The present invention relates to an odorless functional garlic fermented beverage having a tonic effect by natural fermentation. It is known that the main ingredients of garlic, allin and allicin, have excellent physiological effects of garlic, including antibacterial, anticancer, cardio and vascular protective effects, as well as therapeutic effects of various adult diseases.

    However, due to the strong toxicity of raw garlic, as well as garlic's inherent off-flavor (異臭) is even taboo to eat garlic itself.

    Therefore, it is required to develop a functional garlic drink with a gentle fragrance that anyone can drink by removing the strong toxicity of raw garlic and completely eliminating the intrinsic odor of garlic.

   By using the characteristics of garlic, garlic-coffee production method, ocher garlic ring manufacturing method, garlic preparation and garlic beverages with reduced garlic odor, the composition of the ingredients and preparation method of garlic beverage, functional garlic beverage composition and preparation method thereof, garlic Prior arts such as juice preparation methods are known.

   However, in the prior art, in order to remove the strong toxic and adverse odor of raw garlic, other herbal medicines such as boiled, ginger, licorice, goji berry, and mugwort are added, but the odor can be removed and everyone can drink softly. Garlic drink has not been developed.

The present invention is to solve the problems of the prior art, while purifying the strong toxicity of raw garlic and remove the reverse odor (하면서도) while maintaining the physiological activity of garlic intact 110 days natural fermentation process for three times Through this, the fermented beverage for preparing naturally fermented garlic fermented beverage, which completely eliminates toxicity and odor, was prepared.

     The composition of the present invention is 1) what garlic will be used as a raw material, 2) what process will remove toxic and odor completely, 3) what is the biological activity of these naturally fermented garlic beverages Cri / Bgi CD It was developed experimentally through animal experiment of rats and developed odorless, functional garlic fermented beverage with tonic effect by natural fermentation.

1. Selection of Raw Garlic for Garlic Beverage

   There are several kinds of garlic. The effect of temperature on the formation of scales of garlic is large, depending on the varieties, early and late. Hypertrophy of the scaly stem begins around 10 ° C and accelerates around 20 ° C, with different work sensitivities depending on temperature. The Nanji system has low temperature monolithity compared to the Hanji system, and the Hanji system exhibits high temperature uniformity compared to the Nanji system.

   In the present invention, the garlic beverage of the present invention is prepared using the recently popular ground garlic.

2. Manufacturing Process of Garlic Beverage Deodorized

(1) primary fermentation

   The native ground garlic used as a raw material is sorted and washed without peeling, and then mixed with garlic in a mixed solution of sugar-fructose-oligosaccharide (30%: 30%: 40%, w / w) using native pottery The solution is placed in an indigenous clay pot so that the ratio is 1.0: 1.5, and covered with a thin gauze and an earthenware lid, followed by primary fermentation for 25 days.

(2) secondary fermentation

  The fermented extract from which the first naturally fermented garlic was extracted was filtered through a filter and centrifuged at 4,3,500 rpm for 10 minutes at low temperature to store the supernatant as the primary fermentation extract (A), and the dried garlic was dried in a dryer at 30 ° C. 5 days of dried garlic, peeled garlic and sugar-exaggerated-starch syrup-oligosaccharides (25%: 30%: 30%: 15%, w / w) mixed solution 1: 1 Cover with a thin gauze and pot cover, and then ferment for 2 days naturally, and soak the garlic skin in a certain amount of clear water.

(3) tertiary fermentation

  After completion of the second natural fermentation, the fermented extracts from which garlic was removed were filtered through a filter and centrifuged at 4,3,500 rpm for 10 minutes at low temperature to store the supernatant as a secondary fermentation extract (B), and the dried garlic was dried at 30 ° C. Dry for 3 days at.

   The primary fermentation extract (A) and the secondary fermentation extract (B) are mixed at a ratio of 1: 1, and the garlic husks containing the garlic husk are mixed at a ratio of 5: 1, and the dry garlic is soaked. After the injection, finally fermented for 3 days in a plastic house for 3 days to prepare a stock solution of the garlic beverage of the present invention.

   In this manner, three natural fermentations for 110 days remove all the pharmacological and nutrient components of garlic itself to the final fermentation extract, and completely eliminate the strong toxicity and the off-flavor of fresh garlic. Garlic stock fermented for 110 days is soft and fragrant.

3. Evaluation of Physiological Activity of Garlic Fermented Beverages

   Garlic fermentation beverage (GFB) diluted to 15%, 35%, 55%, respectively, using the final tertiary natural fermentation extract of garlic of the present invention, respectively, GFB-15, GFB-35, GFB-55 Cri / Bgi CD rats (180 ± 10 g) were used as the experimental group and were freely administered via water bottles for 6 weeks. The daily dose of rats is considered to be 20 ml.

 1) Effect of Various Cholesterol Contents

  ◇ Determination of total cholesterol content

Total cholesterol content in serum was measured by o -phthaldehyde method according to the method of Rudel et al. (1973). First, 0.1 ml of each sample is taken, 0.3 ml of 33% KOH solution and 3.0 ml of 95% ethanol are added and mixed well. Serum is then heated for 15 minutes and brain cell fractions are heated in a 60 ° C. water bath for 60 minutes and then cooled. 5.0 ml of hexane was added to the mixture, and 3.0 ml of distilled water was added thereto, then mixed well for 1 minute, and the layers were separated to take 1.0 ml of the nucleic acid layer. The hexane layer is concentrated and dried with nitrogen, and 2.0 ml of o- phthaldehyde reagent is added and mixed well. After 10 minutes, 1.0 ml of concentrated sulfuric acid is added and mixed as a coloring reagent. Within 10 to 90 minutes after the addition of sulfuric acid, the absorbance was measured at 550 nm using a spectrophotometer, and the total cholesterol content (mg / dl serum) was quantified by a standard calibration curve.

  ◇ Measurement of lipoprotein-cholesterol content

  HDL, LDL-cholesterol content in serum was measured using HDL-cholesterol (HDL-C 555, Eiken, Japan), LDL-cholesterol (BLF, Eiken, Japan) kit reagent.

   ①. Determination of HDL-Cholesterol Content

  0.3 ml of serum is added to the test tube, and 0.3 ml of the precipitating reagent is mixed well and left for 10 minutes at room temperature, followed by centrifugation at 700 × g for 10 minutes. After that, add 50 ml of the supernatant, 50 µl of standard solution (100 mg / dl), and 50 µl of distilled water to 3.0 µl of distilled water, respectively, mix well, and warm for 5 minutes in a 37 ° C water bath. As a control, the absorbance at 555 nm was used to quantify the content of HDL-cholesterol (mg / dl serum).

  ②. Determination of LDL-Cholesterol Content

  Add 0.1 ml of serum and 0.1 ml of standard serum to the test tube, add 4.0 ml of BLF kit reagents I and II, and mix well for 5 seconds, and then distilled water within 10 minutes after standing at room temperature (25 ± 3 ℃) for 25 minutes. In contrast, the absorbance at 650 nm was measured to quantify the content of LDL-cholesterol (mg / dl serum).

Therefore, the contents of total cholesterol, LDL-cholesterol and HDL-cholesterol in the blood of GFB-15, GFB-35, and GFB-55 groups for 6 weeks in CD rats were treated with distilled water (DW) as a control. In comparison, it is shown in <Table 1>.

As shown in <Table 1>, the effect of garlic fermented beverage (GFB) on total cholesterol was found to effectively reduce the total cholesterol content in blood according to the administration of garlic fermented beverage. In addition, the content of LDL-cholesterol, which is a bad cholesterol known as a cause of chronic degenerative disease, also showed a clear inhibitory effect of more than 15% of LDL-cholesterol in the GFB-35 and GFB-55 administration groups.

   In addition, the content of HDL-cholesterol, which has been found to be good cholesterol as an anti-cholesterol factor or longevity factor, has been found to be higher than 15% of HDL according to the administration of garlic fermented beverage (GFB). It was found that the content of cholesterol was effectively increased. Therefore, as a result of evaluating the effect of garlic fermented beverages on cholesterol content, the GFB-35 experimental group containing 35% garlic fermented beverages can be evaluated as the most effective garlic fermented beverages.

< Table 1 > Effect of Garlic Fermented Beverage (GFB) on the Content of Cholesterol

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 Total Cholesterol Content LDL-Cholesterol Content HDL-Cholesterol Content

              (mg / dl serum) (mg / dl serum) (mg / dl serum)

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Control (7) * 132.30 ± 6.55 ** 75.06 ± 2.81 24.55 ± 2.12

                  100.0% 100.0% 100.0%

GFB-15 administration group (7) 119.50 ± 8.35 68.35 ± 6.35 26.91 ± 2.63

90.3% ^a 91.2% ^a 109.6% ^a

GFB-30 administration group (7) 115.32 ± 11.42 63.77 ± 4.82 28.80 ± 3.02

87.2% ^b 85.0% ^b 117.3% ^c

GFB-55 administration group (7) 118.02 ± 10.52 64.82 ± 4.39 28.40 ± 3.02

89.2% ^a 86.4% ^b 115.6% ^c

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 GFB-15, 35 and 55: 15%, 35% and 55% of garlic fermentation beverage; * Used

Number of rats; ** mean ± standard deviation; Significance test for control: ^a p <0.05; ^b p <0.01; ^c p <0.001

 2) Evaluation of the atherosclerotic index

   The atherosclerotic index (AI), which was identified as the incidence index of adult disease, was calculated by Haglund et al. (1991) by subtracting the HDL-cholesterol content from the total cholesterol content and dividing it by the HDL-cholesterol content.

   Chronic degenerative disease is called adult disease, and when the initial symptoms of these diseases, LDL-cholesterol, which is a bad cholesterol, settle through the walls of the blood vessels, arteriosclerosis begins to harden as the elasticity of the blood vessels decreases. The incidence index of atherosclerosis, which is an early symptom of adult disease, is called the atherosclerotic index (AI) and is widely used as a diagnostic tool for all adult diseases.

Therefore, the effect of garlic fermented beverage (GFB) on the atherosclerosis index is shown in <Table 2>. As shown in Table 2, the garlic fermented beverage (GFB) -administered group had a 20-30% chance of not having an adult disease, indicating that the atherosclerotic index was effectively reduced. Therefore, it is expected that the Garlic fermented beverage can be prevented and prevented by adult diseases.

<Table 2> Effect of Garlic Fermented Beverages on Atherosclerosis Index

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       Atherosclerotic index *

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Control group (7) ** 4.39 ± 0.33 *** 100.0%

GFB-15 administration group (7) 3.44 ± 0.31 78.4% ^c

GFB-35 administration group (7) 3.00 ± 0.06 68.3% ^c

GFB-55 administration group (7) 3.15 ± 0.19 71.7% ^c

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  GFB-15, 35 and 55: 15%, 35% and 55% of garlic fermentation beverage;

  * Atherogenic index = [(Total Cholesterol-HDL-Cholesterol / HDL-Cholesterol] ** Number of rats used;

Mean ± standard deviation; Significance test for the control group: ^c p <0.001

3) Evaluation of Aging Inhibitory Activity

   As a result of animal experiments, it is expected that garlic fermented beverages (GFB) can effectively inhibit aging in vivo, given that garlic fermented beverages have a 20-30% higher probability of suppressing adult diseases. The aging of the living body is the fact that the generation of free radicals (oxygen free radicals) by a variety of causes causing oxidative stress to promote the aging. As a biological defense enzyme that inhibits the aging of the living body, superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), and other free radical scavenger enzymes Exists. We will evaluate the effect of garlic fermented beverage (GFB) on free radicals, oxidative stress and free radical scavenging enzymes.

 ◇ Evaluation of free radicals and oxidative stress

  ① Determination of the content of hydroxy radicals

  Deoxyribose is a method of measuring the degree of hydroxyl radical formation. Deoxyribose is destroyed by reactive oxygen metabolites to generate aldehydes. Measured accordingly.

   Serum hydroxyl radical was first measured by adding 33.3 μl of 0.1 M phosphate buffer solution (pH 7.4), 10 mM NaN3, 7 mM deoxyribose, 5 mM ferrousammonium sulfate, and 0.54 M NaCl. The experimental group combines the sample (15 μl) and water (185 μl) to 200 μl and warms the mixed solution in a 37 ° C. constant temperature water bath for 15 minutes. However, the control group skips the warming process and puts a sample of the control group (30 μl) into the test tube. Add water (185 μl) to all test tubes and mix well. Add 75 μl of 8.1% SDS solution, 500 μl of 20% acetic acid, and 25 μl of water to the reaction solution, and mix well with 333 μl of 1.2% TBA solution. After boiling for 30 minutes and then cooled at room temperature, the supernatant obtained by centrifugation at 800 × g for 5 minutes was measured by using a spectrophotometer to measure the absorbance at 532 nm. The amount of hydroxy radicals (nmole / mg protein / min) was calculated.

  ② Determination of the content of superoxide radicals

The production of superoxide radical was measured by reducing the rate of ferricytochrome C that can inhibit superoxide dismutase according to the method of MaCord et al. (1969) and Chan et al. (1974). That is, 20 mM cyanide solution was added to 420 μl of phosphate buffer solution (pH 7.8) containing 0.1 mM EDTA so that the concentration of cyanide was 50 μM, and then warmed at 37 ° C. for 10 minutes. 300 μl of cytosol and 50 μl of 0.1 mM cytochrome C were added to the solution, and the absorbance at 550 nm was measured over time using a spectrophotometer. At this time, the amount of cytochrome C was calculated with a molecular absorption coefficient of 19,500M ^-1 cm ^-1 .

  ③ Determination of the content of lipid peroxide (LPO)

  Lipid peroxide (LPO) content in serum was measured by TBA method according to the method used by Choi et al. (1990) to determine the content of malondialdehyde (MDA). Take 20 μl of serum and 180 μl of distilled water in each test tube, add 200 μl of 8.1% sodium dodecyl sulfate (SDS) solution for about 5 seconds, add 1.5 ml of 20% acetic acid, and mix well for 5 seconds again. Add 1.0 ml of 1.2% TBA (thiobarbituric acid) reagent, stopper with clean beads, and heat in a water bath for 30 minutes. The reaction solution was centrifuged at 800 x g for 10 minutes, and the supernatant was measured for absorbance at 532 nm using a spectrophotometer to quantify the content of lipid peroxide (nmole / ml serum) according to a standard calibration curve.

 ④ Effect on the generation of free radicals

Oxygen free radicals such as hydroxy radicals (OH), superoixde radicals (O ₂ ⁻ ) and hydrogen peroxide (H ₂ O ₂ ) are all cancers, including cancer. , Aging, and senile dementia. Therefore, when comparing the effect of garlic fermented beverages (GFB) on the production of these active oxygen, as shown in Table 3, the amount of hydroxy radical (OH) is the same in the GFB-30 and GFB-55 administration groups. was recognized by the generation inhibitory effect of the 14% effective · OH radicals, O ₂ · ^- production also has been created inhibitory effect of 14 to 16% was observed in the GFB-30 and GFB-55 treated group of.

  Therefore, long-term use of garlic fermentation beverage (GFB) is expected to be very effective in preventing most adult diseases, cancers of incurable diseases, senile dementia, and aging.

 ⑤ Effect on oxidative stress

Free radicals attack lipid components in vivo to produce lipid peroxide (LPO), attack proteins to form oxidized protein (OP) that forms various crosslinks, and attack nucleic acid to attack 8-OHdG Produces intractable diseases such as cancer and dementia and promotes aging. As shown in Table 3, in the GFB-35 and GFB-55 administration groups as garlic fermented beverages, a very effective inhibitory effect of LPO of 14-15% compared to the control group was recognized. Therefore, long-term use of garlic fermented beverages can effectively prevent the production of lipid peroxide (LPO), a causative agent of aging, and is expected to effectively prevent cancer, including aging, senile dementia and incurable diseases.

< Table 3 > Effect of Garlic Fermented Beverage (GFB) on Free Oxygen and Oxidative Stress

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 Amount of hydroxy radical in the dosing group Content of superoxide radical Content of lipid peroxide

           (nmol / mg protein) (nmol / mg protein) (nmol / mg protein)

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Control Group (7) * 19.10 ± 0.98 ** 45.70 ± 2.41 6.14 ± 0.61

                     100.0% 100.0% 100.0%

GFB-15-administered group (7) 17.60 ± 1.37 41.78 ± 2.41 5.68 ± 0.47

                      92.2% 91.4% 92.5%

GFB-30 administration group (7) 16.43 ± 1.37 38.25 ± 1.87 5.27 ± 0.37

86.1% ^b 83.7% ^c 85.8% ^c

GFB-55 administration group (7) 16.68 ± 0.51 39.25 ± 3.63 5.18 ± 0.19

87.3% ^b 85.8% ^b 84.3% ^c

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 GFB-15, 35 and 55: 15%, 35% and 55% of garlic fermentation beverage; * Used

Number of rats; ** mean ± standard deviation; Significance test for control: ^a p <0.05; ^b p <0.01; ^c p <0.001

 ◇ Activity evaluation of free radical scavenging enzyme

  ① Determination of the activity of superoxide dismutase

   According to the method of Oyanagui et al. (1984), the activity of superoxide dismutase (SOD) is 0.5 ml of distilled water and 0.1 A reagent (52.125) in 0.1 ml of a solution diluted 30-fold with serum phosphate buffer (pH 8.2). Add 0.2 ml of mg of hydoxylamine + 102.1 mg of hypoxanthine / 250 ml DW and 0.2 ml of B reagent (20 μl of 3.75 mM xanthine oxidase + 0.9939 mg ethylene diaminetetraacetic acid / 26.7 ml phosphate buffer, pH 8.2).

   After warming for 40 minutes in a constant temperature water bath at 37 ℃, add 2.0 ml of C reagent (300 mg of sulfanilic acid + 5.0 mg of N-1-naphthylethyene diamine acid / 500 ml of 16.7% acetic acid) and mix and mix for 20 minutes at room temperature. After standing, the absorbance was measured at 550 nm using a spectrophotometer to quantify superoxide dismutase (unit / mg protein) by standard calibration curve.

  ② Determination of the activity of catalase

  Measurement of catalase (CAT) activity in serum by the method of Rigo et al. (1977) was performed in vitro by 250 μl of phosphate buffer solution (130 mM, pH 7.0), 330 μl of distilled water, and 15 mM hydrogen peroxide solution in 20 μl of serum. 900 μl was added, mixed well for 5 seconds, and immediately measured using a spectrophotometer for 2 minutes of change in absorbance over time at 240 nm, and 1 unit was calculated by decomposition of 1 μmol of hydrogen peroxide for 1 minute.

  ③ Effect on scavenger enzyme

Enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT), known as biodefense enzymes, are scattered throughout the body because they have to prevent aging. It has the characteristics. Comparing the administration effect of garlic fermented beverage (GFB) on the activity of these active oxygen scavenging enzymes, as shown in <Table 4>, the GFB-35 and GFB-55 administration groups were 16-20% highly effective SOD. Enzyme activity was increased, and CAT enzyme activity was significantly increased by 10-14% in the GFB-35 and GFB-55 groups.

   Therefore, long-term administration of garlic fermented beverages effectively promotes the activity of SOD and CAT enzymes, which are the most important biological defense enzymes, and protects against various adult diseases and aging, as well as incurable diseases such as cancer and senile dementia through the protective effect of toxic free radicals. It is expected to be very effective prevention.

< Table 4 > Effect of garlic fermented beverage (GFB) on the activity of free radical scavenger

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 Activity of dose group superoxide dismutase (SOD) Activity of catalase (CAT)

                    (unit / mg protein) (μmol / mg protein / min)

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Control (7) * 1.05 ± 0.10 ** 100.0% 46.72 ± 0.70 100.0%

GFB-15 treated group (7) 1.13 ± 0.11 106.9% 48.36 ± 1.53 103.5%

GFB-30 administration group (7) 1.26 ± 0.06 119.8% ^c 51.40 ± 1.58 110.0% ^a

GFB-55 administration group (7) 1.22 ± 0.13 116.1% ^c 53.08 ± 1.87 113.6% ^b

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 GFB-15, 35 and 55: 15%, 35% and 55% of garlic fermentation beverage; * Used

Number of rats; ** mean ± standard deviation; Significance test for control: ^a p <0.05; ^b p <0.01; ^c p <0.001

In the present invention, natural fermentation completely eliminated the toxicity and odor through the 110-day natural fermentation process to purify the strong toxicity of raw garlic and remove the reverse odor (하면서도) while preserving and maintaining the physiological activity of garlic as it is Anyone can easily drink because it has been prepared for the preparation of garlic beverages and has the effect of tonic by the functionality of garlic.

Claims (2)

     30%: 30%: 40% of worted domestic garlic and sugar-fructose-oligosaccharides were washed in native pottery at a ratio of 1.0: 1.5 in native pottery for 25 days and fermented for 25 days. The mixed solution of 25%: 30%: 30%: 15%, w / w garlic and sugar-exaggerated starch-oligosaccharides in which the primary fermentation extract (A) and the dried garlic were dried in a dryer at 30 ° C. for 5 days. In a ratio of 1: 1 to native pottery, and fermented for 2 days in a 40-day natural fermentation, and then mixed with a fermented extract (B) extracted from garlic at a ratio of 1: 1, and garlic peeled with garlic peel The odorless functional garlic fermented beverage having a tonic effect by natural fermentation, characterized in that the mixture was mixed in a ratio of 5: 1, and then fermented for 3 days in a vinyl house for 3 days.      30%: 30%: 40% of worted domestic garlic and sugar-fructose-oligosaccharides were washed in native pottery at a ratio of 1.0: 1.5 in native pottery for 25 days and fermented for 25 days. The mixed solution of 25%: 30%: 30%: 15%, w / w garlic and sugar-exaggerated starch-oligosaccharides in which the primary fermentation extract (A) and the dried garlic were dried in a dryer at 30 ° C. for 5 days. In a ratio of 1: 1 to native pottery, and fermented for 2 days in a 40-day natural fermentation, and then mixed with a fermented extract (B) extracted from garlic at a ratio of 1: 1, and garlic peeled with garlic peel A method of preparing an odorless functional garlic fermented beverage having a tonic effect by natural fermentation, which is prepared by mixing the solution at a ratio of 5: 1, and then tertiary fermentation in a vinyl house for 45 days.