CN111973495A - Preparation method and application of fermented vegetable-berry fruit product - Google Patents
Preparation method and application of fermented vegetable-berry fruit product Download PDFInfo
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- CN111973495A CN111973495A CN201910748702.0A CN201910748702A CN111973495A CN 111973495 A CN111973495 A CN 111973495A CN 201910748702 A CN201910748702 A CN 201910748702A CN 111973495 A CN111973495 A CN 111973495A
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- vegetable
- berry
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- fermented
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Classifications
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- A—HUMAN NECESSITIES
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
- A23L19/09—Mashed or comminuted products, e.g. pulp, purée, sauce, or products made therefrom, e.g. snacks
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Abstract
The invention relates to the field of plant fermentation, in particular to a preparation method and application of a vegetable-berry fruit fermented product. The invention provides a preparation method of a fermented product of vegetable and berry, which comprises the steps of extracting a mixture of vegetable juice and berry juice in a three-stage way to obtain the fermented product of vegetable and berry; wherein the three-stage extraction comprises a first stage aqueous phase extraction step, a second stage alcohol phase extraction step and a third stage organic acid phase extraction step. The invention also provides the vegetable and berry fruit fermentation product prepared by the method and application of the vegetable and berry fruit fermentation product in preparing a composition for improving antioxidant activity. The three-stage extraction process of the invention can improve the total polyphenol content of the mixture of vegetable juice and berry juice and/or the activity of superoxide dismutase.
Description
Technical Field
The invention relates to the field of plant fermentation, in particular to a preparation method and application of a vegetable-berry fruit fermented product.
Background
Free radicals (Free radial) generally refer to reactants whose peripheral orbital region contains unpaired electrons, and are commonly found in Reactive Oxygen Species (ROS), Reactive Nitrogen Species (RNS), and the like; the ROS is a highly reactive oxygen-containing substance, and includes Peroxide (Superoxide), Superoxide (Superoxide), Hydroxyl radical (Hydroxyl radical), Singlet oxygen (Singlet oxygen), and the like.
In human body, the generation of free radicals mostly originates from mitochondria, and when the mitochondria carries out aerobic respiration, a small part of electrons escaped from an electron transfer chain of an inner membrane of the mitochondria react with oxygen to form free radicals such as superoxide, peroxide and the like. Free radicals participate in many physiological mechanisms in the human body, including fertilization, maturation, cytokine production, elimination of toxic substances, etc., and play an important role in combating bacterial and viral invasion and cancer; in contrast, however, excessive free radicals can produce oxidative stress in individuals, and free radicals are known to be associated with the occurrence of many diseases that affect the normal function of the body, possibly leading to aging and death of the body. And with the change of living habits, the modern human body content is easy to generate excessive free radicals, which causes great threat to health.
The natural vegetables and fruits contain a lot of antioxidants, however, in order to actually achieve the antioxidant effect of the human body, a large amount of vegetables and fruits must be eaten, and through the cooking and digestion processes, a large amount of antioxidant substances in the vegetables and fruits are lost, so that the antioxidant effect is greatly reduced; therefore, there is a need to develop a method for increasing the content of antioxidant substances in vegetables and fruits and improving the antioxidant activity of the vegetables and fruits.
Disclosure of Invention
Accordingly, an object of the present invention is to provide a method for preparing a fermented product of berries and vegetables, comprising extracting a mixture of juice and juice in a three-stage process to obtain the fermented product; wherein the three-stage extraction comprises a first stage aqueous phase extraction step, a second stage alcohol phase extraction step, and a third stage organic acid phase extraction step.
The present invention also provides a fermented product of vegetable and fruit, wherein the fermented product is prepared from a mixture of vegetable juice and berry juice by the above-mentioned preparation method.
Another object of the present invention is to provide a use of the above mentioned fermented vegetable and berry fruit for preparing a composition for improving antioxidant activity.
In one embodiment of the present invention, the first stage of aqueous phase extraction step is to extract the mixture of vegetable juice and berry juice with water as solvent at 50-100 ℃ for 0.5-3 hours to obtain a product of the first stage of aqueous phase extraction step, and the mixture of vegetable juice and berry juice and water are mixed in a ratio of 1:1-5 (w/w); the second stage alcohol phase extraction step and the third stage organic acid phase extraction step are implemented by fermenting the product of the first stage aqueous phase extraction step with yeast (Saccharomyces cerevisiae) and lactic acid bacteria (Streptococcus thermophilus) at 25-35 deg.C for 1-10 days, and the addition amount of the yeast is 0.01-0.5% (w/w); the addition amount of the lactobacillus is 0.01-0.25% (w/w).
In another embodiment of the present invention, the aforementioned three-stage extraction method of the present invention is to increase the total polyphenol content in the fermented vegetable-berry fruit product of the present invention.
In another embodiment of the present invention, the aforementioned three-stage extraction method of the present invention is to increase the activity of superoxide dismutase in the fermented product of berries and fruits of the present invention.
The three-stage extraction method comprises the steps of firstly, separating water-soluble trace elements and mineral substances from raw materials in a first stage of water phase extraction; the second stage alcohol phase extraction step is to further extract volatile and low-polarity substances; and finally, the third stage of organic acid phase fermentation step is to extract the produced organic acid phases such as lactic acid and the like through yeast and lactic acid bacteria fermentation, and separate the organic acid and the weak electrolyte substances. By the three-stage extraction, the essence in the mixture raw material of the vegetable juice and the berry juice is completely extracted, so as to achieve the purposes of increasing effective components and improving the efficacy of the raw material.
The three-stage extraction method can obviously improve the content of total polyphenol and the activity of superoxide dismutase in the mixture of the vegetable juice and the berry juice, can effectively extract water-soluble mineral substances, trace elements, low-polarity substances and organic compounds in the raw materials of the mixture of the vegetable juice and the berry juice, achieves the purposes of refining molecules and improving functional components, and can enhance the health-care and anti-oxidation effects of the raw materials. In addition, the fermented vegetable and berry product obtained by the three-stage extraction method can be used for preparing a composition for improving antioxidant activity, and the composition is a medicine, a health-care product or a food and can be administered to an individual by oral administration, smearing and the like.
The following examples are presented to illustrate the present invention and are not to be construed as limiting the scope of the invention, which is intended to be limited only by the appended claims.
Detailed Description
As used herein, the numerical values are approximations and all numerical data are reported to be within the 20 percent range, preferably within the 10 percent range, and most preferably within the 5 percent range.
Statistical analysis was performed using Excel software. Data are presented as mean ± Standard Deviation (SD), and differences between individual data are analyzed by student's t-test (student's t-test).
As used herein, the term "mixture of vegetable and berry juice" means a mixed juice of vegetable and berry juice, wherein the vegetable and berry juice are mixed in a weight ratio of 1: 1.
As used herein, the term "fermented vegetable-berry product" means a mixture of vegetable-berry juice and an extraction solvent of water, which are uniformly mixed in a ratio of 1:1-5(w/w), and then subjected to a first stage of aqueous phase extraction for a specific time and temperature, and then to a second stage of alcohol phase extraction and a third stage of organic acid phase fermentation by using yeast and lactic acid bacteria, and finally obtained by the three-stage extraction method. Wherein the yeast is strain BCRC20271, and the Lactobacillus plantarum is strain BCRC 910636.
The procedures and parameters involved in the extraction according to the present invention are within the ordinary skills and routine skill of those skilled in the art.
The procedures and parameters for the fermentation of microorganisms according to the present invention are within the skill of those skilled in the art.
According to the present invention, the drug may be manufactured in a dosage form suitable for parenteral (parenteral) or topical (topologic) administration using techniques well known to those skilled in the art, including, but not limited to: injections (injection) [ e.g., sterile aqueous solution (sterile aqueous solution) or dispersion (dispersion) ], sterile powders (sterile powder), external preparations (external preparation), and the like.
According to the present invention, the pharmaceutical may further comprise a pharmaceutically acceptable carrier (pharmaceutically acceptable carrier) which is widely used in pharmaceutical manufacturing technology. For example, the pharmaceutically acceptable carrier may comprise one or more agents selected from the group consisting of: solvents (solvent), buffers (buffer), emulsifiers (emulsifying), suspending agents (suspending agent), disintegrating agents (disintegrant), disintegrating agents (disintegrating agent), dispersing agents (dispersing agent), binding agents (binding agent), excipients (excipient), stabilizers (stabilizing agent), chelating agents (chelating agent), diluents (diluent), gelling agents (gelling agent), preservatives (preserving), wetting agents (wetting agent), lubricants (lubricating), absorption delaying agents (absorption delaying agent), liposomes (liposome) and the like. The selection and amounts of such agents are within the skill and routine skill of those skilled in the art.
According to the present invention, the pharmaceutically acceptable carrier comprises a solvent selected from the group consisting of: water, normal saline (normal saline), Phosphate Buffered Saline (PBS), aqueous alcohol-containing solutions (aqueous solution linking alcohol), and combinations thereof.
According to the invention, the medicament may be administered by a parenteral route (parenteral routes) selected from the group consisting of: subcutaneous injection (subecanal injection), intradermal injection (intraepithelial injection), and intralesional injection (intralesion).
According to the present invention, the external preparation is prepared by mixing the medicine of the present invention with a base (base) as well known to those skilled in the art.
According to the invention, the substrate may comprise one or more additives (additives) selected from the following group: water, alcohols, glycols, hydrocarbons such as petroleum jelly and white petrolatum]Wax (wax) [ such as Paraffin and yellow wax (yellow wax) ]Preserving agents (preserving agents), antioxidants (antioxidants), surfactants (surfactants), absorption enhancers (absorption enhancers), stabilisers (stabilizing agents), gelling agents (gelling agents) [ such as carbopol [ ]
Microcrystalline cellulose (microcrystalline cellulose) and carboxymethyl cellulose (carboxymethyl cellulose)]Active agents (actives), moisturizers (humectants), odor absorbers (odor absorbers), perfumes (fragrans), pH adjusting agents (pH adjusting agents), chelating agents (chelating agents), emulsifiers (emulsifiers), occlusive agents (occlusive agents), softeners (emulsifiers), thickeners (thickeners), solubilizing agents (solubilizing agents), penetration enhancers (penetration enhancers), anti-irritants (anti-irritants), colorants (colorants), and propellants (propellants). The selection and amounts of these additives are within the skill of the artAre well within the expertise and routine of those skilled in the art.
According to the present invention, the care product may further comprise an acceptable adjuvant (acceptable adjuvant) which is widely used in the art of care product manufacture. For example, the acceptable adjuvant may comprise one or more agents selected from the group consisting of: solvents, gelling agents, active agents, preservatives, antioxidants, screening agents, chelating agents, surfactants, colouring agents, thickening agents, fillers, fragrances and odour absorbers. The selection and amounts of such agents are within the skill and routine skill of those skilled in the art.
In accordance with the present invention, the cosmetic may be manufactured in a form suitable for skin care (skincare) or makeup (makeup) using techniques well known to those skilled in the art, including, but not limited to: aqueous solutions (aqueous solutions), aqueous-alcoholic solutions (aqueous-alcoholic solutions) or oily solutions (oil solutions), emulsions in the form of oil-in-water type, water-in-oil type or compound type, gels, ointments, creams, masks (masks), patches, wipes, powders, aerosols, sprays, lotions, serums, pastes, foams, dispersions, drops, mousses (mousses), sunblocks, lotions (toiletries), foundations (foundations), make-up removal products (make-up removal products), soaps (soaps) and other body cleansing products (body cleansing products), and the like.
In accordance with the present invention, the cosmetic may also be used in combination with one or more known active topical agents (external use agents) selected from the following: whitening agents (whitening agents) [ such as retinoic acid (tretinoin), catechins (catechin), kojic acid, arbutin and vitamin C ], moisturizers, anti-inflammatory agents (anti-inflammatory agents), bactericides (bacteriodes), ultraviolet absorbers (ultraviolets absorbers), plant extracts [ such as aloe vera extract (aloe vera extract) ], skin nutrients (skin nutrients), anesthetics (anesthesics), anti-acne agents (anti-acne agents), antipruritics (antipruritics), analgesics (analgesics), anti-dermatitis agents (antipermatitis agents), anti-hyperkeratotic agents (anti-hypercholesterolitic agents), anti-dry skin agents (anti-dry skin agents), anti-perspirants (anti-perspirant agents), anti-aging agents (anti-aging agents), anti-wrinkle agents (anti-rinking agents), anti-seborrheic agents (anti-anerrheic agents), wound healing agents (wound-healing agents), corticosteroids (corticosteriods), and hormones (hormones). The selection and amounts of such agents for external use are within the skill and routine skill of those skilled in the art.
According to the present invention, the food product may be used as a food additive (food additive) to be added during the preparation of the raw material or during the preparation of the food by conventional methods, and formulated with any edible material into a food product for ingestion by humans and non-human animals.
According to the present invention, the types of food products include, but are not limited to: beverages (leafages), fermented foods (fermented foods), bakery products (bakery products), health foods (health foods) and dietary supplements (dietary supplements).
The invention provides a three-stage extraction method capable of simultaneously achieving molecule refinement and functional component improvement and application of a fermented product of vegetable and berry fruits obtained by the method to improve antioxidant activity. Wherein the yeast is strain BCRC20271, and the Lactobacillus plantarum is strain BCRC 910636. The vegetable and berry fruit fermentation product can obviously improve the content of total polyphenol in the mixture of vegetable juice and berry juice and the activity of superoxide dismutase in the mixture.
Meanwhile, the composition for improving the antioxidant activity of the invention can also comprise an effective amount of the vegetable-berry fruit fermentation product and a pharmaceutically acceptable carrier, and the composition is a medicine, a health-preserving product or a food.
The following detailed three-stage extraction preparation method of the fermented vegetable and berry product, the test for improving the efficacy of the total polyphenol in the fermented vegetable and berry product by the three-stage extraction method, and the test for improving the efficacy of the superoxide dismutase activity in the fermented vegetable and berry product by the three-stage extraction method will be described in detail, so as to confirm that the total polyphenol content in the fermented vegetable and berry product and the activity of the superoxide dismutase in the fermented vegetable and berry product can be significantly improved by the three-stage extraction method.
Example 1 three-stage extraction preparation method of fermented vegetable and berry product of the present invention
In an embodiment of the present invention, a mixture of a vegetable juice and a berry juice (mixed in a weight ratio of 1: 1) and an extraction solvent of water are uniformly mixed in a ratio of 1:1-5(w/w) to perform a first-stage aqueous phase extraction, the mixture is simultaneously sterilized and extracted at 50-100 ℃ for 0.5-3 hours, the crude extraction solution is cooled to room temperature for a subsequent second-stage alcohol phase extraction and a third-stage organic acid phase extraction fermentation step, 0.01-0.5% (w/w) of a yeast (Saccharomyces cerevisiae, purchased from center for biological resources preservation and research, taiwan, No. BCRC20271) and 0.01-0.25% (w/w) of a lactic acid bacterium are simultaneously inoculated into the crude extraction solution, wherein the present invention uses Streptococcus (Streptococcus thermophilus TCI 125, deposited in taiwan, 23 days 6 month 2014, numbered BCRC 910636; the strain has been deposited in Taiwan patent publication No. I519644), and is fermented at 25-35 deg.C for 1-10 days, with the actual time varying according to the fermentation state; wherein, metabolite (alcohol compound) generated by yeast is used for alcohol phase extraction, and metabolite (organic acid compound) generated by lactobacillus is used for organic acid phase extraction. And finally, under the condition that the two bacteria are not removed, using the set specification of sugar degree range 2-4 degrees, pH <4, alcohol > 5% and the like, and if the specification is detected to be in accordance with the specification, judging that the fermentation is finished and obtaining the fermentation liquor. Then, carrying out reduced pressure concentration on the fermentation liquor at 45-70 ℃, filtering by using a 200-400mesh sieve, adding 20-40% isomalto-oligosaccharide to adjust the specification, heating at 95-105 ℃ for 60-90 minutes to carry out sterilization, and obtaining the raspberry and fruit fermented product; wherein the vegetable juice and berry juice are purchased from Yangya marketing limited company, the names of the vegetable juice and berry juice are respectively comprehensive vegetable and fruit concentrated juice and comprehensive berry concentrated juice, and the product numbers are MX-AC134-FL and MX-AC130-FC respectively; the commercially available comprehensive vegetable and fruit concentrated juice is concentrated juice of white grapes, apples, carrots, sugarcanes, passion fruits, pineapples, lemons, green cauliflowers, celery and asparagus, and no additional water is added; the commercially available comprehensive berry concentrated juice is concentrated juice of blueberry, red grape, white grape, waxberry, cranberry and mulberry, and no additional water is added.
The three-stage extraction method comprises the steps of firstly, separating water-soluble trace elements and mineral substances from raw materials in a first stage of water phase extraction; the second stage alcohol phase extraction step is to further extract volatile and low-polarity substances; and finally, the third stage of organic acid phase fermentation step is to extract the produced organic acid phases such as lactic acid and the like through yeast and lactic acid bacteria fermentation, and separate the organic acid and the weak electrolyte substances. By the three-stage extraction, the essence in the mixture raw material of the vegetable juice and the berry juice is completely extracted, so as to achieve the purposes of increasing effective components and improving the efficacy of the raw material.
Example 2 the three-stage extraction method of the present invention improves the efficacy of total polyphenols in the fermented vegetable and berry product of the present invention
In one embodiment of the present invention, in order to perform a test experiment for improving the efficacy of total polyphenols in the fermented vegetable and berry fruit product of the present invention by the three-stage extraction method of the present invention, the Folin-Ciocalteu colorimetric method is used to determine the total polyphenol content in the substitute, and the content is compared with the fermented vegetable and berry fruit product (a wild herb ferment liquid purchased from sphenoidal co., japan, number Lot #160314) of other brands on the market; the determination is carried out by utilizing the antioxidant property of polyphenol, Phosphotungstic acid (Phosphotungstic acid) and Phosphomolybdic acid (phosphomolybdydic acid) in the reagent are reduced by polyphenol (Mo is reduced by Mo 6+To Mo5 +) This then results in the formation of blue compounds (absorbance at 750 nm) whose shade is proportional to the total polyphenol content and which can therefore be used for the quantification of generationsMeasuring the content of total polyphenols in the product.
First, Gallic acid (purchased from Sigma, USA, No. G7384) was used as a standard to prepare a calibration curve, 10.0G of Gallic acid was precisely weighed and placed in a 10mL volumetric flask, and ddH was added thereto2O to a total volume of 10mL and completing a Gallic acid standard (Gallic acid stock, 1000. mu.g/mL), which was then diluted 10-fold first to a concentration of 100. mu.g/mL (100. mu.L Gallic acid stock + 900. mu.L ddH)2O, unused Gallic acid stock stored at-20 deg.C), then 100. mu.g/mL of Gallic acid was diluted in series to 0, 20, 40, 60, 80, and 100. mu.L/mL of Gallic acid (as shown in Table one), 100. mu.L of each concentration of the standard solution was put into a 10mL glass tube, 500. mu.L of Folin-Ciocalteu phenol reagent (available from Merck, Germany, No. 1.09001.0100) was added thereto, mixed well, and left to stand for 3 minutes, 400. mu.L of 7.5% Sodium carbonate (Sodium carbonate, 7.5g of anhydrous Sodium carbonate was quantified to 100mL of ddH 2O, wherein the anhydrous sodium carbonate is purchased from Sigma, usa under the number 31432), is uniformly mixed and then is left to react for 30 to 60 minutes, preferably 30 minutes, and is left to react by shaking (Vortex) mixing until no bubble is determined, 200 μ L of each tube of the reaction solution is taken out and placed in a 96-well culture plate, and the absorbance at 750nm is measured to draw a regression curve formula of the standard solution.
TABLE I formulation for series dilution of gallic acid standard
Then, 100 μ L of other brands of fermented vegetable and fruit and the fermented vegetable and fruit of the present invention are respectively put into a 10mL glass test tube, 500 μ L of Folin-Ciocalteu phenol reagent are respectively added and mixed evenly and are kept still for 3 minutes, 400 μ L of 7.5% sodium carbonate is added and is kept still for 30-60 minutes, preferably 30 minutes, and then oscillation (Vortex) is used for mixing and keeping still until no bubble is determined, 200 μ L of each tube reaction solution is taken out and put into a 96-hole culture plate, the light absorption value at 750nm is measured, the concentration is calculated by the regression curve formula of the standard solution by an interpolation method, and then the dilution times are multiplied to obtain the original fermented vegetable and fruit of other brands and the concentration of total polyphenol in the fermented vegetable and fruit of the present invention.
The test results of the three-stage extraction method for improving the efficacy of the total polyphenol in the fermented vegetable-berry fruit product are shown in table two. The fermented product of the rubus fujiensis of other factories only contains 52.5 mu g/mL of total polyphenol, but the total polyphenol content of the fermented product of the rubus fujiensis of the invention is up to 395.3 mu g/mL, which is 7.5 times of that of other factories. The results show that the three-stage extraction method can significantly improve the content of total polyphenols in the mixture of vegetable juice and berry juice, so as to achieve the purposes of improving functional components and enhancing health-care antioxidant efficacy.
Second, the total polyphenol content in the fermented product of vegetable and berry fruit of the invention and the fermented product of vegetable and berry fruit of other factories
| Total polyphenols (μ g/mL) | |
| The invention relates to a vegetable-berry fruit fermented product | 395.3 |
| Fermented product of vegetable, berry and fruit of Tacheng brand | 52.5 |
Example 3 the three-stage extraction method of the present invention improves the activity of superoxide dismutase in fermented vegetable-berry products of the present invention
In an embodiment of the present invention, in order to perform a test experiment for improving the efficacy of Superoxide dismutase (SOD) activity in the fermented product of rubus suavissimus of the present invention by the three-stage extraction method, the test is performed with reference to the criterion of determining the activity of Superoxide dismutase in the GB/T5009.171-2003 health food, and the above-mentioned fermented product of rubus suavissimus of other manufacturers on the market is also used for comparison, wherein the SOD activity is quantified by the efficiency of the substance to be tested for inhibiting the self-oxidation reaction of Pyrogallol. SOD, an enzyme that catalyzes superoxide through disproportionation and converts superoxide into oxygen and hydrogen peroxide, is widely found in various animals, plants, and microorganisms, and is an important antioxidant in the body to protect cells exposed to oxygen.
Firstly, preparing solution A and solution B for measurement, wherein solution A is prepared by dissolving 1.2114g of Tris (hydroxymethyl) aminomethane (Tris) and 37.2mg of ethylene diamine tetraacetic acid (EDTA-2Na) in 62.4mL of 0.1mole/L hydrochloric acid (HCl) solution, and adding distilled water to 100mL after dissolving, and solution B is prepared by dissolving 56.7mg of pyrogallol in a small amount of hydrochloric acid with a concentration of 10mmole/L, and adding pure water to 100mL after dissolving.
Treating a sample to be tested, wherein the solid sample is prepared by the following method: weighing 1g of the solid sample, placing the solid sample in a mortar, adding 9mL of pure water, grinding and dissolving, transferring all the liquid into a centrifugal tube, washing the mortar with a small amount of pure water, then adding the residual liquid into the centrifugal tube, quantifying the residual liquid with the pure water until the total volume is 10mL, centrifuging the liquid at 4000rpm for 15 minutes, and taking the supernatant for determination. The liquid samples were prepared as follows: the clarified liquid sample can be directly measured as a stock solution; the turbid liquid sample was centrifuged at 4000rpm for 5 minutes and the supernatant was taken for assay. The liquid sample can be diluted properly with pure water and finally multiplied by the dilution factor to calculate the original concentration.
Next, a blank assay for the control group was performed: adding 2.35mL of the solution A, 2mL of pure water and 0.15mL of the solution B into a centrifuge tube in sequence, adding the solution B, mixing, quickly and gently shaking, mixing uniformly, pouring into a colorimetric tube immediately, measuring the absorbance at 325nm, reacting for 1 minute, and detecting the absorbance at 325nm The difference between the two values is Δ A235(ii) a Respectively adding 2.35mL of the solution A, 1.8mL of pure water, 0.2mL of the sample to be detected (vegetable and fruit fermentation products of other factories or vegetable and fruit fermentation products of the invention) and 0.15mL of the solution B into a centrifuge tube, mixing, quickly shaking and uniformly mixing, immediately pouring into a colorimetric tube to determine the light absorption value at 325nm, reacting for 1 minute, detecting the light absorption value at 325nm, wherein the difference value of the two values is delta A235(as shown in table three below).
Table III, SOD activity determination measuring table
| Reagent | Blank space | Sample (I) |
| Solution A | 2.35mL | 2.35mL |
| Pure water | 2mL | 1.8mL |
| Sample to be tested | － | 0.2mL |
| Solution B | 0.15mL | 0.15mL |
Finally, the SOD activity in the fermented product of the rubus and the fruit of other factories and the fermented product of the rubus and the fruit of the invention is calculated by the following formula, wherein V is 4.5, and D is 0.2.
ΔA325: auto-oxidation speed of pyrogallol
ΔA′325: sample inhibition of pyrogallol autoxidation speed
V: volume of sample
D: dilution factor of sample
The test results of the effect of the three-stage extraction method for improving the activity of superoxide dismutase in the fermented vegetable-berry fruit are shown in table four. The activity of superoxide dismutase in the fermented product of the rubus fujiensis of the other factories is 1153U/mL of total polyphenol content, and the activity of superoxide dismutase in the fermented product of the rubus fujiensis of the invention is 1657U/mL which is 1.4 times of that of the fermented product of the other factories. The results show that the three-stage extraction method can obviously improve the activity of superoxide dismutase in the mixture of vegetable juice and berry juice so as to achieve the purposes of improving functional components and enhancing health-care antioxidant effect.
TABLE IV SOD Activity in the present invention and other Brand fermented products of vegetable and berry fruit
| SOD(U/mL) | |
| The invention relates to a vegetable-berry fruit fermented product | 1657 |
| Tare vegetable-berry fruit fermentationArticle (A) | 1153 |
In conclusion, the three-stage extraction method for preparing the mixture of the vegetable juice and the berry juice can obviously improve the content of total polyphenol and the activity of superoxide dismutase in the mixture of the vegetable juice and the berry juice, and can effectively extract water-soluble minerals, trace elements, low-polarity substances and organic compounds in the raw materials of the mixture of the vegetable juice and the berry juice, thereby simultaneously achieving the purposes of refining molecules and improving functional components and enhancing the health-care and anti-oxidation effects of the raw materials. In addition, the fermented vegetable and berry product obtained by the three-stage extraction method can be used for preparing a composition for improving antioxidant activity, and the composition is a medicine, a health-care product or a food and can be administered to an individual by oral administration, smearing and the like.
Claims (10)
1. A preparation method of fermented product of vegetable and fruit comprises extracting a mixture of vegetable and fruit juice with a three-stage extraction process to obtain fermented product; wherein the three-stage extraction comprises a first stage aqueous phase extraction step, a second stage alcohol phase extraction step and a third stage organic acid phase extraction step.
2. The method of claim 1, wherein the first aqueous phase extraction step comprises extracting the mixture of vegetable and berry juices with water at 50-100 deg.C for 0.5-3 hr to obtain the product of the first aqueous phase extraction step.
3. The method of claim 2, wherein the mixture of vegetable and berry juice is mixed with water in a ratio of 1:1-5 (w/w).
4. The method according to claim 2, wherein the second alcohol phase extraction step and the third organic acid phase extraction step are steps of fermenting the product of the first aqueous phase extraction step with a yeast and a lactic acid bacteria.
5. The method of claim 4, wherein the step of fermenting is carried out at 25-35 ℃ for 1-10 days.
6. The method according to claim 4, wherein the yeast is added in an amount of 0.01-0.5% (w/w); the addition amount of the lactobacillus is 0.01-0.25% (w/w).
7. A fermented vegetable and fruit product, wherein the fermented vegetable and fruit product is prepared from a mixture of vegetable and fruit juice by the preparation method of claim 1.
8. The fermented product according to claim 7, wherein the preparation method is to increase the total polyphenol content in the fermented product.
9. The fermented product according to claim 7, wherein the preparation method is to increase the activity of superoxide dismutase in the fermented product.
10. Use of a fermented vegetable-berry fruit according to claim 7 for the preparation of a composition for increasing antioxidant activity.
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| US201862676396P | 2018-05-25 | 2018-05-25 | |
| TW108118146A TWI699208B (en) | 2018-05-25 | 2019-05-24 | Integrated fruits and vegetables and berries ferment, preparation method thereof and use thereof |
| TW108118146 | 2019-05-24 |
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| CN111973495A true CN111973495A (en) | 2020-11-24 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115804739A (en) * | 2021-09-15 | 2023-03-17 | 百岳特生物技术（上海）有限公司 | Fermented product of autumn plum, and its preparation method and application |
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| CN115804739A (en) * | 2021-09-15 | 2023-03-17 | 百岳特生物技术（上海）有限公司 | Fermented product of autumn plum, and its preparation method and application |
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| TWI699208B (en) | 2020-07-21 |
| TW202011978A (en) | 2020-04-01 |
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