CN113304229A - Fermentation-based conditioning preparation for hypertension and hyperlipidemia and application thereof - Google Patents

Fermentation-based conditioning preparation for hypertension and hyperlipidemia and application thereof Download PDF

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Publication number
CN113304229A
CN113304229A CN202110599724.2A CN202110599724A CN113304229A CN 113304229 A CN113304229 A CN 113304229A CN 202110599724 A CN202110599724 A CN 202110599724A CN 113304229 A CN113304229 A CN 113304229A
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parts
fermentation
black
tea
seed
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张建欣
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Luoyang Furuhao Chuanhe Technology Co ltd
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Luoyang Furuhao Chuanhe Technology Co ltd
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    • A23LFOODS, 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
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/88Liliopsida (monocotyledons)
    • A61K36/899Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • A61K36/8998Hordeum (barley)
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00Drugs for disorders of the metabolism
    • A61P3/06Antihyperlipidemics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00Drugs for disorders of the cardiovascular system
    • A61P9/10Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00Drugs for disorders of the cardiovascular system
    • A61P9/12Antihypertensives
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10Preparation or pretreatment of starting material
    • A61K2236/19Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment

Abstract

The invention relates to a fermentation-based conditioning preparation for hypertension and hyperlipidemia and application thereof, belonging to the field of fermentation, wherein the conditioning preparation is prepared by fermenting raw materials with fermentation strains; the feedstock comprises a variety of categories such as: vegetables (garlic, balsam pear, etc.), fruits (pineapple, apple, etc.), tea (vine tea, etc.), Chinese herbal medicines (salvia miltiorrhiza, radix puerariae, etc.), and the fermentation strains are saccharomyces cerevisiae, lactobacillus pentosus and lactobacillus japonicas. The enzyme preparation is prepared by adopting a multi-bacterium fermentation process and based on multi-class mixed raw materials without toxic and side effects, the raw materials used for fermentation are easy to obtain, the cost is low, the content of nutrient components in fermentation liquor is rich, the oxidation resistance is strong, and the enzyme preparation has the effects of improving blood pressure and blood fat, cholesterol, myocardial infarction and cerebral infarction.

Description

Fermentation-based conditioning preparation for hypertension and hyperlipidemia and application thereof
Technical Field
The invention belongs to the technical field of fermentation, and particularly relates to a preparation for improving hypertension and hyperlipidemia based on fermentation and application thereof.
Background
The serum of healthy people contains various lipoproteins at a certain concentration, and the person with blood fat or lipoprotein level often exceeding the normal limit is called hyperlipidemia or hyperlipoproteinemia. Hyperlipidemia is the basis of atherosclerosis, and high-density lipoprotein can transport cholesterol to liver for decomposition, so that the hyperlipidemia has an anti-atherosclerosis effect. Atherosclerosis is prone to develop isolated systolic hypertension. On the contrary, in the hyperlipidemia patients with hypertension, due to the damage of the intima of the blood vessel, blood fat is easy to enter the media of the artery, and arteriosclerosis is formed. Therefore, hypertension and hyperlipidemia are always classified as risk factors of atherosclerosis or coronary heart disease in the medical field. Blood lipid levels are affected by a variety of factors, including dietary structure, lifestyle, and a variety of diseases or drugs. The hypertension is often caused by genetic factors and is related to high-salt diet living in cold regions, long-term oral antihypertensive drug treatment is needed, complications are prevented and treated, hyperlipidemia is caused by weakness and dizziness, patients are obese, and the patients are related to diet greasiness and lack of movement. Some hypotensive drugs affect the side effects of lipid metabolism, so careful selection of drugs is required for patients with hyperlipidemia.
Disclosure of Invention
In view of the above problems, the invention aims to provide a conditioning preparation for hypertension and hyperlipidemia based on fermentation, and aims to provide an application of the conditioning preparation in preparation of health care products or special meal foods for lowering blood pressure and blood lipid. The conditioning preparation is a multi-bacterium fermentation stock solution, is prepared by combining and fermenting saccharomyces cerevisiae, lactobacillus pentosus and lactobacillus japonicas, has easily obtained raw materials for fermentation, low cost and rich contents and varieties of nutrients and functional components in fermentation liquor, and has the function of improving blood pressure, blood fat, cholesterol, myocardial infarction or cerebral infarction.
In order to achieve the purpose, the invention adopts the specific scheme that:
the fermentation-based conditioning preparation for hypertension and hyperlipidemia is prepared by fermenting raw materials with fermentation strains; the method specifically comprises the following steps:
step one, pretreatment of raw materials: drying and grinding the raw materials, and sieving to prepare a mixed material;
the raw materials comprise the following components in parts by weight: 1-3 parts of purple grapes, 3-7 parts of pineapples, 3-7 parts of grapefruit, 1-5 parts of apples, 1-3 parts of olive fruits, 1-3 parts of lemons, 1-5 parts of saussurea involucrate, 1-3 parts of kiwi fruits, 1-3 parts of blueberries, 1-3 parts of black medlar, 3-7 parts of hawthorn fruits, 1-3 parts of pickled muskmelon, 1-5 parts of iron yams, 1-3 parts of purple onions, 3-7 parts of black garlic, 1-5 parts of bitter gourds, 8-12 parts of black tartary buckwheat, 1-3 parts of black fungus, 1-3 parts of chickpeas, 8-12 parts of walnut kernels, 1-5 parts of black beans, 1-5 parts of almonds, 1-5 parts of red yeast rice, 8-12 parts of black peanuts, 8-12 parts of gynostemma pentaphylla, 8-12 parts of lotus leaf tea, 8-12 parts of black sesame seeds, and the like, 3-7 parts of vine tea, 1-5 parts of cyclocarya paliurus tea, 1-5 parts of pine pollen, 3-7 parts of Hangzhou white chrysanthemum, 1-5 parts of snow chrysanthemum, 8-12 parts of frosted mulberry leaf tea, 3-7 parts of plantain herb, 1-5 parts of honeysuckle, 1-5 parts of crocodile tea, 1-5 parts of pseudo-ginseng, 1-5 parts of apocynum venetum, 1-3 parts of raw cassia seed, 3-7 parts of saffron, 1-5 parts of dandelion, 1-5 parts of astragalus root, 1-5 parts of ginkgo seed, 1-5 parts of cordyceps militaris, 8-12 parts of bitter melon seed, 1-5 parts of salvia miltiorrhiza, 8-12 parts of radix puerariae, 1-5 parts of Korean ginseng, 1-5 parts of celery seed, 8-12 parts of American ginseng, 1-5 parts of angelica, 1-5 parts of gastrodia elata, 1-5 parts of grapefruit seed, 1-5 parts of sea buckthorn, 1-5 parts of chrysanthemum flower, and the like, 1-5 parts of five-year dried orange peel, 8-12 parts of corn stigma, 1-5 parts of dendrobium, 1-5 parts of selenium malt, 1-5 parts of cornus officinalis and 1-5 parts of peony seeds.
Step two, adding 20-40g of carbon source and 80-100g of the mixed material obtained in the step one into each liter of water, uniformly mixing, sterilizing and preparing a fermentation substrate;
step three, inoculating fermentation strains into the fermentation substrate obtained in the step two, wherein the fermentation conditions are as follows: anaerobic fermentation at 30 deg.C and 200rpm for 20 days at pH of 4.5-6.5; the fermentation strain is as follows: mixing saccharomyces cerevisiae, lactobacillus pentosus and lactobacillus japonicas according to the volume ratio of 1:1: 1; the inoculum size was 3%.
As a further optimization, in the first step, the raw materials comprise the following components in parts by weight: 1 part of purple grape, 3 parts of pineapple, 3 parts of grapefruit, 1 part of apple, 1 part of olive fruit, 1 part of lemon, 2 parts of snow lotus, 1 part of kiwi fruit, 1 part of blueberry, 1 part of black wolfberry, 3 parts of hawthorn, 1 part of Ma bubble melon, 2 parts of iron yam, 1 part of purple onion, 3 parts of black garlic, 2 parts of balsam pear, 8 parts of black tartary buckwheat, 1 part of black fungus, 1 part of chickpea, 8 parts of walnut kernel, 2 parts of black bean, 2 parts of almond, 2 parts of red kojic rice, 8 parts of black peanut, 8 parts of gynostemma pentaphylla, 8 parts of lotus leaf tea, 8 parts of black sesame, 3 parts of vine tea, 2 parts of cyclocarya paliurus tea, 2 parts of pine pollen, 3 parts of white Hangzhou chrysanthemum, 2 parts of snow chrysanthemum, 8 parts of frosted mulberry leaf tea, 3 parts of plantain herb, 2 parts of honeysuckle, 2 parts of alligator tea, 2 parts of pseudo-ginseng, 2 parts of apocynum venetum, 1 part of raw cassia seed, 3 parts of saffron, 2 parts of dandelion, 2 parts of astragalus root, 2 parts of cordyceps, 2 parts of Chinese caterpillar fungus, 8 parts of white atractylodes, 8 parts of bitter gourd seed, 2 parts of bitter gourd fruit and the like, 2 parts of salvia miltiorrhiza, 8 parts of radix bupleuri and radix puerariae, 2 parts of alpine ginseng, 2 parts of celery seed, 8 parts of American ginseng, 2 parts of angelica, 2 parts of gastrodia elata, 2 parts of grapefruit seed, 2 parts of sea buckthorn, 2 parts of five-year dried orange peel, 8 parts of corn stigma, 2 parts of dendrobium, 2 parts of selenium malt, 2 parts of cornus officinalis and 2 parts of peony seed.
As a further optimization, step two in the preparation of the fermentation substrate, 40g of glucose and 80g of the batch obtained in step one per liter of water were added.
The invention also provides application of the conditioning preparation in preparing health-care products for reducing blood pressure and blood fat or special meal foods.
Has the advantages that:
the invention has various raw material varieties, and not only contains daily food, but also comprises medicinal and edible Chinese herbal medicines. The daily food contains a large amount of bioactive components, but the daily food is usually ignored by people and treated by expensive medicines when treating diseases, and no matter western medicines or pure Chinese herbal medicines can cause organ damage due to invisible pressure on a human body after being taken. According to a great deal of practice, the invention adopts daily food and Chinese herbal medicine to be mixed, and a specific process is adopted for fermentation, so that the fermentation stock solution has nutrition and functional compounds, is more in variety and content, and has less damage to human bodies compared with pure traditional Chinese medicines. The effective substances in the fermentation liquor comprise vitamins, phenols, minerals, functional oligosaccharides, enzymes and other secondary metabolites. The results show that the composition contains more than 200 substances, wherein the types of flavonoid compounds are the most, the proportion of amino acid and derivatives thereof is the highest, and in addition, organic acids, phenolic acids, alkaloids, nucleotides and derivatives thereof are also main components. In addition, the fermentation liquor has stronger free radical scavenging capacity, wherein the hydroxyl free radical, DPPH and ABTS and the superoxide anion VCEAC are respectively 1.54mg/ml, 85.9mg/ml, 2.69mg/ml and 0.92mg/ml, and the fermentation liquor shows excellent oxidation resistance. The traditional Chinese medicine composition can be particularly applied to the regulation of blood pressure and blood fat reduction of middle-aged and elderly people, and the practical use condition shows that the traditional Chinese medicine composition can effectively improve and prevent blood pressure and blood fat, cholesterol, myocardial infarction or cerebral infarction.
Drawings
FIG. 1 is a graph of a Vc standard in a total reducing power assay;
FIG. 2 is a graph of Vc standard in total antioxidant capacity determination;
FIG. 3 is a graph of Vc standard in DPPH radical scavenging assay;
FIG. 4 is a graph of the effect of fermentation broth addition on DPPH radical clearance;
FIG. 5 is a graph of the Vc standard in ABTS free radical clearance determination;
FIG. 6 is a graph of the effect of fermentation broth addition on ABTS radical clearance;
FIG. 7 is a graph of the Vc standard in superoxide anion removal determinations;
FIG. 8 is a graph of the effect of fermentation broth addition on superoxide anion removal;
FIG. 9 is a graph of Vc standard in hydroxyl radical scavenging assay;
FIG. 10 is a graph showing the influence of the amount of a fermentation liquid added on the hydroxyl radical scavenging rate.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention.
Example 1
Firstly, a fermentation process raw material formula: 1 part of purple grape, 3 parts of pineapple, 3 parts of grapefruit, 1 part of apple, 1 part of olive fruit, 1 part of lemon, 2 parts of snow lotus, 1 part of kiwi fruit, 1 part of blueberry, 1 part of black wolfberry, 3 parts of hawthorn, 1 part of Ma bubble melon, 2 parts of iron yam, 1 part of purple onion, 3 parts of black garlic, 2 parts of balsam pear, 8 parts of black tartary buckwheat, 1 part of black fungus, 1 part of chickpea, 8 parts of walnut kernel, 2 parts of black bean, 2 parts of almond, 2 parts of red kojic rice, 8 parts of black peanut, 8 parts of gynostemma pentaphylla, 8 parts of lotus leaf tea, 8 parts of black sesame, 3 parts of vine tea, 2 parts of cyclocarya paliurus tea, 2 parts of pine pollen, 3 parts of white Hangzhou chrysanthemum, 2 parts of snow chrysanthemum, 8 parts of frosted mulberry leaf tea, 3 parts of plantain herb, 2 parts of honeysuckle, 2 parts of alligator tea, 2 parts of pseudo-ginseng, 2 parts of apocynum venetum, 1 part of raw cassia seed, 3 parts of saffron, 2 parts of dandelion, 2 parts of astragalus root, 2 parts of cordyceps, 2 parts of Chinese caterpillar fungus, 8 parts of white atractylodes, 8 parts of bitter gourd seed, 2 parts of bitter gourd fruit and the like, 2 parts of salvia miltiorrhiza, 8 parts of radix bupleuri and radix puerariae, 2 parts of alpine ginseng, 2 parts of celery seed, 8 parts of American ginseng, 2 parts of angelica, 2 parts of gastrodia elata, 2 parts of grapefruit seed, 2 parts of sea buckthorn, 2 parts of five-year dried orange peel, 8 parts of corn stigma, 2 parts of dendrobium, 2 parts of selenium malt, 2 parts of cornus officinalis and 2 parts of peony seed, wherein one part is one or two.
Pretreatment: drying and grinding the raw materials at 55 ℃, and then sieving the raw materials with a 80-mesh sieve to prepare a mixed material.
Fermentation substrate: 40g glucose +80g mixed material +1000mL purified water.
And (3) sterilization conditions: sterilizing at high temperature and high pressure, and keeping the temperature at 121 ℃ for 10 min.
Fermentation strain: saccharomyces cerevisiae + Lactobacillus pentosus + Lactobacillus saxifragi; the three strains are respectively cultured until the logarithmic phase, and then mixed according to the volume ratio of 1:1:1 to prepare the mixed strain. The strains were all purchased commercially.
Inoculation amount: 3 percent of mixed bacteria inoculation.
Fermentation conditions are as follows: 30 ℃, 200rpm, 20 days, natural pH (5.3 +/-0.3), anaerobic fermentation.
Second, component detection and analysis
1. The liquid chromatography and mass spectrometry are used for qualitative and quantitative analysis of the substance components in the multi-bacterium fermentation stock solution prepared by the fermentation process, and at least 263 substances exist in the fermentation stock solution, wherein the maximum types of flavonoid compounds are 54, the second types of amino acids and derivatives thereof are 34, and the highest ratio of the amino acids and the derivatives thereof is 21.70%. The composition of the substances contained therein was summarized and the results are shown in table 1 below.
Table 1: the species and the ratio of the substance components in the fermentation stock solution.
Figure BDA0003092472550000051
Figure BDA0003092472550000061
Figure BDA0003092472550000071
2. Determination of Total reducing force
2.1 principle of experiment (Potassium ferricyanide method)
The measurement of the reducing power is essentially a process of checking whether or not a substance is a good electron donor. The electrons supplied by the highly reducing species can react with the free radicals to form relatively inert species, in addition to reducing the oxidizing species, to interrupt the autoxidative chain reaction and render the free radicals stable.
With Prussian blue Fe4[Fe(CN)6]3The amount of production was used as an index. Potassium ferricyanide K3Fe(CN)6Reduction to potassium ferrocyanide, Fe2+Further ferric chloride (FeCl)3) And prussian blue formation. The 700nm absorbance reflects the generation amount of Prussian blue, and the greater the absorbance value is, the stronger the total reducing force value of the sample is, which indicates that the better the antioxidation effect is.
2.2 Experimental methods:
firstly, diluting a fermentation stock solution to 5% (V/V) by using a phosphate buffer solution (0.2mol/L, pH6.6);
② 2.5mL of the diluent is added with 2.5mL of 1% potassium ferricyanide (W/V). Shaking, and water bath at 50 deg.C for 30 min;
③ rapidly cooling, adding 2.5mL of 10 percent trichloroacetic acid (W/V), shaking up, 3000r/min, and centrifuging for 10 min;
fourthly, taking 2.5mL of supernatant, adding 2.5mL of deionized water and 0.5mL of 0.1 percent ferric chloride (W/V), shaking up the mixture, and carrying out water bath at 25 ℃ for 10 min;
measuring the light absorption value at 700nm by using deionized water as a reference;
and sixthly, taking Vc as a positive control.
2.3 results of the experiment
The 2.3.1Vc standard curve is shown in fig. 1.
2.3.2 fermentation stock solution total reducing power equivalent Vc amount.
A700 of a 5% dilution of the fermentation broth was found to be 1.296, which was substituted into the following equation:
Figure BDA0003092472550000081
in the above formula, VCEAC-equivalent Vc amount, absorbance at A-700nm, and B-dilution factor.
The VCEAC obtained was 1.160 mg/mL.
3. Total antioxidant capacity determination
3.1 principle of experiment (phosphomolybdic method)
The principle of the phosphorus molybdenum complex assay is that Mo (VI) is reduced to a green Mo (V) complex by an antioxidant substance, and the maximum absorption wavelength is 695 nm. The stronger the antioxidant activity, the greater the absorbance value measured.
3.2 Experimental methods:
1mL of fermentation stock solution diluent (10%, (0.2mol/L, pH6.6) prepared by phosphate buffer solution);
adding 3mL of reaction solution (0.6mol/L sulfuric acid, 28mmol/L sodium phosphate and 4mmol/L ammonium molybdate), oscillating and shaking uniformly, and carrying out water bath in a water bath kettle at the temperature of 95 ℃ for 90 min;
cooling to room temperature, using deionized water as reference, and measuring absorbance at 695 nm;
and thirdly, Vc is used as a positive control.
3.3 results of the experiment.
The 3.3.1Vc standard curve is shown in fig. 2.
3.3.2 the total antioxidant capacity of the fermentation stock solution is equivalent to Vc amount.
A695 was measured as 0.519 for a 10% dilution of the fermentation broth, which was substituted into the following equation:
Figure BDA0003092472550000082
in the above formula, VCEAC-equivalent Vc amount, light absorption value at A-695nm and B-dilution multiple.
VCEAC was obtained at 0.538 mg/mL.
4 determination of the radical clearance.
4.1 DPPH radical scavenging rate.
4.1.1 experimental principle:
DPPH free radical scavenging ability is one of the most stable antioxidant indexes at present, and the DPPH free radical and a free radical scavenger are combined in an electron pairing mode to lighten the color of the whole system.
4.1.2 Experimental methods:
taking 0.2-2.0mg/mL, 0.2mg/mLVc of n, 0.2-2% and 2mL of 0.2% of fermentation stock solution of n, adding 4mL of 0.1mmol/L DPPH-ethanol solution, adding 450uL of Tris-HCl of pH, shaking up by shaking, carrying out water bath in a dark place at 25 ℃ for 30min, and taking deionized water as reference, wherein A is 517nm and A is 1;
replacing a sample with 2mL of deionized water at 517nm, A0;
③ to replace DPPH-ethanol with 4mL ethanol, 517nm, A2;
Figure BDA0003092472550000091
4.1.3 results of the experiment
4.1.3.1 Vc standard curve is shown in FIG. 3.
4.1.3.2 DPPH clearance rate EC of fermentation stock solution50And an equivalent Vc. The broth DPPH clearance curve is shown in fig. 4. VCEAC 85.9mg/ml, EC500.564%. Wherein: VCEAC is the equivalent vitamin C content (mg) per ml of fermentation broth; EC (EC)50The amount (%) of the fermentation liquid added was determined so that the radical scavenging rate was 50%.
4.2ABTS radical clearance
4.2.1 principle of experiment:
the maximum absorption wavelength of ABTS radical ions was 734nm, so ABTS radical ion concentration was measured under A734 nm. If A734nm decreases, it indicates that ABTS free radicals are scavenged.
4.2.2 Experimental methods:
taking 0.01-0.03mg/mL, 0.002mg/mLVc of n and 2mL of 0.2-2.0% stock solution, adding 8mL of ABTS working solution, uniformly mixing by oscillation, and carrying out water bath reaction at 25 ℃ for 6min, 731nm and A1;
2mL of phosphate buffer solution with the pH value of 7.40.2 mol/L is used for replacing the sample, the concentration is 734nm, and A0 is added;
③ 8mL of phosphate buffer with pH 7.40.2 mol/L is used to replace ABTS, 734nm, A2.
Figure BDA0003092472550000101
4.2.3 results of the experiment
4.2.3.1Vc standard curve is shown in FIG. 5.
4.2.3.2 fermentation stock ABTS clearance EC50And an equivalent Vc. The clearance graph is shown in fig. 6. VCEAC 2.69mg/ml, EC50=0.89%。
4.3 superoxide anion scavenging
4.3.1 principle of the experiment (pyrogallol method)
Under the alkaline condition (pH8.2), pyrogallol undergoes autoxidation reaction, and in the autoxidation process, O is continuously released2-·,O2-The autoxidation process can be further promoted to produce colored intermediates, which are continuously oxidized to other intermediates. The scavenger can scavenge superoxide anion and inhibit O2-Formation of s, which hinders the process of auto-oxidation of pyrogallol.
4.3.2 Experimental methods:
taking 0.05-0.50mg/mL, taking n as 0.05mg/mLVc, 10-100%, taking n as 10% stock solution and 1mL, adding 5mL of 0.05mol/LpH8.2 Tris buffer solution, adding 0.2mL of 12mmol/L pyrogallol after shaking and mixing uniformly, carrying out water bath reaction at 25 ℃ for 5min, 320nm and A1;
replacing a sample with 1mL of deionized water at 320nm, A0;
and thirdly, 0.1mL of deionized water is used for replacing pyrogallol with the concentration of 320nm and A2.
Figure BDA0003092472550000111
4.3.3 results of the experiment
4.3.3.1Vc standard curve is shown in FIG. 7.
4.3.3.2 superoxide anion removal rate EC for fermentation broth50And an equivalent Vc. The clearance graph is shown in fig. 8. VCEAC 0.92mg/ml, EC50=30.67%。
4.4 hydroxyl radical scavenging ratio (salicylic acid method)
4.4.1 principle of the experiment:
hydroxyl radical H generation by Fenton reaction2O2+Fe2+→→[·OH+OH-+Fe3+]Salicylic acid is added into the reaction system, and hydroxyl free radical and salicylic acid antisense generate dihydroxybenzoic acid with special absorption at 510 nm. If an analyte having a hydroxyl radical scavenging function is added to the reaction system, the produced dihydroxybenzoic acid is reduced, and the absorption value at 510nm is also reduced accordingly.
4.4.2 methods of experiment:
firstly, taking Vc and 1mL of fermentation stock solution with certain concentration, adding 1mL of 0.006mol/L ferrous sulfate, then adding 1mL of 0.012mol/L hydrogen peroxide, oscillating and uniformly mixing, carrying out water bath at 37 ℃ for 30min, then adding 1mL of 0.02mol/L salicylic acid-ethanol, oscillating and uniformly mixing, carrying out water bath at 37 ℃ for 30min, and carrying out treatment at 510nm A1;
replacing a sample with 2mL of deionized water at 510nm, A0;
and thirdly, replacing the reaction liquid with 2mL of deionized water, replacing salicylic acid with 1mL of ethanol, and carrying out treatment at 510nm and A2.
Figure BDA0003092472550000112
4.4.3 results of the experiment
4.4.3.1 Vc standard curve is shown in FIG. 9.
4.4.3.2 clearance rate of hydroxyl radical in fermentation stock solution EC50And an equivalent Vc. The clearance graph is shown in fig. 10. VCEAC 1.54mg/ml, EC50≈13.25%
5. Summary of antioxidant capacity of fermentation broth (hypoglycemic)
Various diseases of human beings have been confirmed to be associated with active oxygen and free radicals, such as inflammation, arteriosclerosis, aging, diabetes, cancer, etc., and excessive free radicals in the body cause denaturation of proteins, damage and death of cells, and even pathological changes of the body. The edible ferment contains abundant nutrient substances and other bioactive substances, and has two sources, namely all components of the used raw materials and new components generated after microbial fermentation conversion, and the content of a plurality of bioactive substances is increased to a certain extent through microbial fermentation. Researches show that effective substances in the edible ferment comprise vitamins, phenols, minerals, functional oligosaccharides, enzymes and other secondary metabolites and the like.
The antioxidative effect of the food ferment is realized by removing free radicals in vivo. The reason why the enzyme has the antioxidant effect is as follows: firstly, polyphenol, flavone and other components contained in the raw materials can remove free radicals; second, superoxide dismutase (SOD) and secondary metabolites produced by microbial fermentation can improve the free radical scavenging ability of enzymes and alleviate lipid peroxidation.
Various free radicals are generated in the process of human body metabolism, wherein superoxide anion free radicals (O2- ·) and hydroxyl free radicals (· OH) have the greatest harm to human bodies, can react with any molecule in living cells to cause tissue cytopathy, cause various diseases and cause organism aging; the DDPH method and the ABTS method are commonly used in vitro antioxidant capacity detection methods. In addition, the stronger the reducing power, the stronger the antioxidant power, so the antioxidant effect is mainly judged by the free radical scavenging power and reducing power of the enzyme.
As shown in table 2. The in vitro experiment result shows that the fermentation liquor (for reducing blood sugar) has certain scavenging capacity to superoxide anion free radical (O2- ·), the equivalent Vc is 0.920mg/mL, and the EC50 is 30.670%; the compound has certain scavenging capacity on hydroxyl free radicals (. OH), the scavenging rate is 50 percent under the addition of 13.25 percent of fermentation stock solution, and the equivalent Vc of the fermentation stock solution per milliliter is 1.54 milligrams; this should be a combined action of active ingredients such as superoxide dismutase (SOD), phenols, benzophenones, sterosides and polysaccharides contained in the fermentation broth. The fermentation stock solution has stronger reducing power and higher scavenging capacity for the synthesized free radicals DPPH and ABTS, which indicates that the total antioxidant capacity of the fermentation stock solution is stronger.
Table 2: and (4) analyzing the antioxidation effect of the fermentation stock solution.
Figure BDA0003092472550000131
It should be noted that the above-mentioned embodiments illustrate rather than limit the scope of the invention, which is defined by the appended claims. It will be apparent to those skilled in the art that certain insubstantial modifications and adaptations of the present invention can be made without departing from the spirit and scope of the invention.

Claims (4)

1. The fermentation-based conditioning preparation for hypertension and hyperlipidemia is prepared by fermenting raw materials with fermentation strains; the method is characterized in that: the method specifically comprises the following steps:
step one, pretreatment of raw materials: drying and grinding the raw materials, and sieving to prepare a mixed material;
the raw materials comprise the following components in parts by weight: 1-3 parts of purple grapes, 3-7 parts of pineapples, 3-7 parts of grapefruit, 1-5 parts of apples, 1-3 parts of olive fruits, 1-3 parts of lemons, 1-5 parts of saussurea involucrate, 1-3 parts of kiwi fruits, 1-3 parts of blueberries, 1-3 parts of black medlar, 3-7 parts of hawthorn fruits, 1-3 parts of pickled muskmelon, 1-5 parts of iron yams, 1-3 parts of purple onions, 3-7 parts of black garlic, 1-5 parts of bitter gourds, 8-12 parts of black tartary buckwheat, 1-3 parts of black fungus, 1-3 parts of chickpeas, 8-12 parts of walnut kernels, 1-5 parts of black beans, 1-5 parts of almonds, 1-5 parts of red yeast rice, 8-12 parts of black peanuts, 8-12 parts of gynostemma pentaphylla, 8-12 parts of lotus leaf tea, 8-12 parts of black sesame seeds, and the like, 3-7 parts of vine tea, 1-5 parts of cyclocarya paliurus tea, 1-5 parts of pine pollen, 3-7 parts of Hangzhou white chrysanthemum, 1-5 parts of snow chrysanthemum, 8-12 parts of frosted mulberry leaf tea, 3-7 parts of plantain herb, 1-5 parts of honeysuckle, 1-5 parts of crocodile tea, 1-5 parts of pseudo-ginseng, 1-5 parts of apocynum venetum, 1-3 parts of raw cassia seed, 3-7 parts of saffron, 1-5 parts of dandelion, 1-5 parts of astragalus root, 1-5 parts of ginkgo seed, 1-5 parts of cordyceps militaris, 8-12 parts of bitter melon seed, 1-5 parts of salvia miltiorrhiza, 8-12 parts of radix puerariae, 1-5 parts of Korean ginseng, 1-5 parts of celery seed, 8-12 parts of American ginseng, 1-5 parts of angelica, 1-5 parts of gastrodia elata, 1-5 parts of grapefruit seed, 1-5 parts of sea buckthorn, 1-5 parts of chrysanthemum flower, and the like, 1-5 parts of five-year dried orange peel, 8-12 parts of corn stigma, 1-5 parts of dendrobium, 1-5 parts of selenium malt, 1-5 parts of cornus officinalis and 1-5 parts of peony seeds;
step two, adding 20-40g of carbon source and 80-100g of the mixed material obtained in the step one into each liter of water, uniformly mixing, sterilizing and preparing a fermentation substrate;
step three, inoculating fermentation strains into the fermentation substrate obtained in the step two, wherein the fermentation conditions are as follows: anaerobic fermentation at 30 deg.C and 200rpm for 20 days at pH of 4.5-6.5; the fermentation strain is as follows: mixing saccharomyces cerevisiae, lactobacillus pentosus and lactobacillus japonicas according to the volume ratio of 1:1: 1; the inoculum size was 3%.
2. The conditioning formulation of claim 1 characterized by: in the first step, the raw materials comprise the following components in parts by weight: 1 part of purple grape, 3 parts of pineapple, 3 parts of grapefruit, 1 part of apple, 1 part of olive fruit, 1 part of lemon, 2 parts of snow lotus, 1 part of kiwi fruit, 1 part of blueberry, 1 part of black wolfberry, 3 parts of hawthorn, 1 part of Ma bubble melon, 2 parts of iron yam, 1 part of purple onion, 3 parts of black garlic, 2 parts of balsam pear, 8 parts of black tartary buckwheat, 1 part of black fungus, 1 part of chickpea, 8 parts of walnut kernel, 2 parts of black bean, 2 parts of almond, 2 parts of red kojic rice, 8 parts of black peanut, 8 parts of gynostemma pentaphylla, 8 parts of lotus leaf tea, 8 parts of black sesame, 3 parts of vine tea, 2 parts of cyclocarya paliurus tea, 2 parts of pine pollen, 3 parts of white Hangzhou chrysanthemum, 2 parts of snow chrysanthemum, 8 parts of frosted mulberry leaf tea, 3 parts of plantain herb, 2 parts of honeysuckle, 2 parts of alligator tea, 2 parts of pseudo-ginseng, 2 parts of apocynum venetum, 1 part of raw cassia seed, 3 parts of saffron, 2 parts of dandelion, 2 parts of astragalus root, 2 parts of salvia miltiorrhiza, 8 parts of radix bupleuri and radix puerariae, 2 parts of alpine ginseng, 2 parts of celery seed, 8 parts of American ginseng, 2 parts of angelica, 2 parts of gastrodia elata, 2 parts of grapefruit seed, 2 parts of sea buckthorn, 2 parts of five-year dried orange peel, 8 parts of corn stigma, 2 parts of dendrobium, 2 parts of selenium malt, 2 parts of cornus officinalis and 2 parts of peony seed.
3. The conditioning formulation of claim 1 characterized by: step two in the preparation of the fermentation substrate, 40g of glucose and 80g of the blend obtained in step one were added per liter of water.
4. Use of the conditioning preparation according to any one of claims 1 to 3 in the preparation of a health care product for lowering blood pressure and blood lipid or a special meal food.
CN202110599724.2A 2021-05-31 2021-05-31 Fermentation-based conditioning preparation for hypertension and hyperlipidemia and application thereof Pending CN113304229A (en)

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