CN107660671A - A kind of water chestnut skin drinks - Google Patents

A kind of water chestnut skin drinks Download PDF

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Publication number
CN107660671A
CN107660671A CN201710996358.8A CN201710996358A CN107660671A CN 107660671 A CN107660671 A CN 107660671A CN 201710996358 A CN201710996358 A CN 201710996358A CN 107660671 A CN107660671 A CN 107660671A
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water chestnut
extract
water
raw material
phyllanthus embical
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谷毅鹏
杨小梅
段振华
罗杨合
商飞飞
熊裕豪
谢微
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Hezhou University
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Hezhou University
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23L2/38Other non-alcoholic beverages
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • 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
    • A23L2/00Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52Adding ingredients
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • 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
    • A23L2/00Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/70Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
    • A23L2/84Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • 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
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    • A23L33/105Plant extracts, their artificial duplicates or their derivatives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/335Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • AHUMAN NECESSITIES
    • 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
    • A61K36/02Algae
    • A61K36/03Phaeophycota or phaeophyta (brown algae), e.g. Fucus
    • AHUMAN NECESSITIES
    • 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)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/28Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A61K36/288Taraxacum (dandelion)
    • AHUMAN NECESSITIES
    • 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)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/39Convolvulaceae (Morning-glory family), e.g. bindweed
    • AHUMAN NECESSITIES
    • 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)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/47Euphorbiaceae (Spurge family), e.g. Ricinus (castorbean)
    • AHUMAN NECESSITIES
    • 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)
    • A61K36/89Cyperaceae (Sedge family)
    • 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

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Abstract

The present invention provides a kind of water chestnut skin drinks, it is characterised in that includes the raw material of following composition by weight:Water chestnut bark extract 97 122, Phyllanthus embical fruit extract 23 41, purple sweet potato anthocyanin 5 10, sodium carboxymethylcellulose 59, xylitol 16 25, citric acid 13 19, malic acid 16 28, dandelion extract 22 38, fucoidan 9 17, water 100 120.The present invention is using water chestnut skin as main raw material(s), it is primary raw material therefrom to extract water chestnut skin active component, compound other functional components, it can the functional beverage of strengthen immunity and internal antibacterial anti-inflammatory ability, the present invention be effectively unique flavor, color and luster and composite plant type beverage that is good in taste and being worth with better nutritivity to provide a kind of long-term use.

Description

A kind of water chestnut skin drinks
Technical field
The invention belongs to technical field of functional drink, and in particular to a kind of water chestnut skin drinks.
Background technology
Water chestnut also known as horseshoe, Di Li, black taro, Waternut Herb etc., monocot genus Cyperaceae, scientific name HeleocharisTuberoseSchult, it is perennial root herbaceous plant, originates in SOUTHERN CHINA and India, China is certainly Record has been cultivated during the Western Han Dynastry.Water chestnut it is nutritious, according to analysis:0.8-1.5g containing protein in per the fresh bulbs of 100g, carbon Hydrate 12.9-21.8g, fatty 0.3g, crude fibre 0.3g, ash content 0.8g, calcium 4mg, phosphorus 45mg, iron 0.8mg and a small amount of Carrotene and vitamin C.Water chestnut is not only fresh and sweet tasty, and its bulb also has certain medical value, seedling seedling, root, fruit It is used as medicine.The traditional Chinese medical science thinks that water chestnut is cold in nature, sweet, nontoxic, have heat-clearing, promote the production of body fluid, whet the appetite, helping digestion, resolving sputum, moisturize, improving eyesight, voiceless sound, Sober up and other effects.Water chestnut skin contains abundant polysaccharide, and polysaccharide has anti-oxidant, anti-aging, hypoglycemic, antitumor, immunological regulation Function etc., therefore cause people greatly to pay close attention to.
According to incompletely statistics, the existing water chestnut cultivated area in China is more than 500,000 mu, and gross annual output amount is more than 800,000 tons, Er Qieyou Ascendant trend.Data shows that water chestnut pericarp accounts for the 26% of total water chestnut quality, and so annual caused water chestnut skin is more than 210,000 tons. Now, water chestnut processing enterprise of city of south china has developed to 5,80,000 tons of water chestnut of year digestion, produces water chestnut skin more than 20,000 tons.With The development of water chestnut industry, the quantity of water chestnut pericarp also can be further up.Due to the special nutritive value of water chestnut and health care valency Value, its product category are a lot.Including chufa preserved fruit, water chestnut crisp chip, water chestnut boiled, Water chestnut cake, water chestnut sugar, water chestnut beverage, Water chestnut fruit squash etc..These products will remove water chestnut pericarp in production and processing, caused a large amount of water chestnut skins it is general all by Directly throw away or as animal feed, so water chestnut skin can not sufficiently be utilized.
Therefore, it is necessary to develop a kind of new way of water chestnut skin higher value application.
The content of the invention
In view of this, the present invention provides a kind of water chestnut skin drinks, and the present invention is using water chestnut skin as main raw material(s), therefrom It is primary raw material to extract water chestnut skin active component, compounds other functional components, there is provided a kind of long-term use can effectively increase Strong immunity and the in vivo functional beverage of antibacterial anti-inflammatory ability, the present invention are unique flavor, color and luster and good in taste and tool There is the composite plant type beverage that better nutritivity is worth.
The technical scheme is that:A kind of water chestnut skin drinks, it is characterised in that include the original of following composition by weight Material:Water chestnut bark extract 97-122, Phyllanthus embical fruit extract 23-41, purple sweet potato anthocyanin 5-10, sodium carboxymethylcellulose 5-9, Xylitol 16-25, citric acid 13-19, malic acid 16-28, dandelion extract 22-38, fucoidan 9-17, water 100-120.
Further, the water chestnut skin drinks include the raw material of following composition by weight:Water chestnut bark extract 105- 115th, Phyllanthus embical fruit extract 28-35, purple sweet potato anthocyanin 6-8, sodium carboxymethylcellulose 6-8, xylitol 19-22, citric acid 14-17, malic acid 19-26, dandelion extract 27-34, fucoidan 11-15, water 110-115.
Further, the water chestnut skin drinks include the raw material of following composition by weight:Water chestnut bark extract 110, oil Sweet seed extract 33, purple sweet potato anthocyanin 7, sodium carboxymethylcellulose 7, xylitol 21, citric acid 16, malic acid 22, Pu Public English extract 32, fucoidan 14, water 112.
In the present invention, water chestnut has certain medical value in itself, in addition a variety of natural active products contained in water chestnut skin Matter has been found.Research has shown that compared with conventional chemical preservative, water chestnut bark extract has that antibacterium ability is stronger, Antimycotic ability is weaker, still has stronger antibacterial efficacy in weakly acidic pH matrix environment, and have stronger heat endurance Deng antibacterial characteristics;The ability of the removing free radical of water chestnut bark extract is quite notable, and its antioxidation activity is with the increase of concentration And strengthen;Containing compounds such as Polyphenols and flavonoids in water chestnut bark extract, there is certain scavenging ability of DPPH free radical, Good dose-effect relationship is shown between its Scavenging activity and extract concentrations.
Phyllanthus embical fruit also known as emblic, it is the sweet perennial deciduous tree of category of Euphorbiaceae oil in the present invention.Yunnan Province is national wild Raw Phyllanthus embical fruit is distributed most wide, yield highest province.Count according to investigations, Yunnan Province ten thousand mu of Phyllanthus embical fruit Linda 60-70 in blocks at present, Suitable dough product produces nearly ten thousand tons of fresh fruit per year up to more than 3,000,000 mu.So huge resource is not utilized largely at present, is not had more Have carry out profound comprehensive process-.In recent years, China's medical field and scientific research department were by repetition test and constantly research, card Bright Phyllanthus embical fruit has obvious anti-aging effects;Have to synthesis of the strong carcinogen N- nitroso compounds in animal and human body bright Aobvious blocking effect, blocking rate is up to more than 90.11%.In addition, Phyllanthus embical fruit contains substantial amounts of Vc, carbohydrate, organic acid, list Rather, also containing protein, fat, vitamin B1, vitamin B2, vitamin A, carrotene, cellulose, pectin, alkaloid and Calcium, phosphorus, iron, potassium, sodium etc., there is clearing heat and detoxicating, relieving sore-throat resolving sputum, promote the production of body fluid to quench thirst, solution is tired of the effect of sobering up, and it is treated to abscess of throat Imitate especially pronounced.
In the present invention, Dandelion composite family herbaceos perennial, taraxol, dandelion are contained in dandelion plant A variety of health-nutrition compositions such as element, choline, organic acid, synanthrin, there is broad-spectrum antibacterial effect, to Diplococcus pneumopniae, meningitis ball Bacterium, corynebacterium diphtheriae, Pseudomonas aeruginosa, shigella dysenteriae, typhoid bacillus etc. have a stronger bacteriostatic activity, the dandelion extraction in the present invention Thing has excellent antibacterial modulability.
In the present invention, the algal polysaccharide sulfate is a kind of sulfated polysaccharides extracted from brown alga, mainly there is brown alga Polysaccharide and sulfate composition, are α-L-fucose -4- sulfuric esters that height 3- is branched (1-2) or (1-3) connection.Brown alga is more Sugar sulfate, cytotoxin can be produced by activating macrophage, suppress tumor cell proliferation and kill tumour cell;Separately Tumour growth can also be suppressed by suppressing Tumor Angiongesis outside, can also directly suppress growth of tumour cell.Study table Bright, brown algal polysaccharide sulfate can decline the matrix of cancer cell and homogeneity adhesion, and cell separation rate enhancing, cell passes through Basilar memebrane reduced capability.That is, algal polysaccharide sulfate can make the malignant phenotype of cell change, make its transfer ability It is suppressed.In addition, algal polysaccharide sulfate can increase sensitiveness of the cancer cell to chemotherapeutics.Separately there is research table Bright, the generation of angiocardiopathy is often higher relevant with blood fat in blood and cholesterol level.Algal polysaccharide sulfate can be by chyme In fat take out of in vitro, there is the effect of good lipid-loweringing, norcholesterol, and without the side effect of fat-reducing medicament.Brown alga Polysaccharide sulfate can effectively reduce human body artery systolic pressure, and energy is gentle, is effectively reduced the systolic pressure of patients with hypertension and relaxes Open pressure.Algal polysaccharide sulfate can be as the secondary buck medicine of high blood pressure.Particularly, the algal polysaccharide sulphur in sea-tangle source Acid esters has the bioactivity such as good anticoagulation, antitumor, antiviral, anti-inflammatory, anti-oxidant and immunological regulation, to kidney and liver It is dirty to have protective effect.Fucoidan in the present invention can be prepared by this area any prior art.
In the present invention, in food storage or process, due to the result of bacterial action, from protein or Amino acid score Solution, mostly in such as amine, ammonia, the material of nitrogen-containing heterocycle compound bad flavor of alkalescence, using citric acid and malic acid Distribution, the effect for neutralizing and de-tasting can be reached.
In the present invention, contain pigment in the water chestnut bark extract, Phyllanthus embical fruit extract in itself, then answer with purple sweet potato anthocyanin With combination, assign beverage of the present invention good color and luster.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20(w/w), 0.1%-0.6% pectase is added by the mass ratio of raw material, 10-60min is digested at 45 DEG C; Destroy the enzyme treatment is carried out, filtering, leaves and takes filtrate.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, by the original quality of raw material than adding 0.5%-1.2% pectases, reached using lemon acid for adjusting pH 3.5, digest 15-35min at 50 DEG C, after enzymolysis, enzymolysis liquid is boiled into 30 min enzyme deactivations, is cooled to room temperature, with the Buddhist nun of 200 mesh Imperial cloth filtering, then filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane, obtain the Phyllanthus embical fruit extract solution of clarification.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25(m/v)Feed intake, 100 in water-bath DEG C h of refluxing extraction 2, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion Extract dry powder.
Particularly, the purple sweet potato anthocyanin in the present invention is dry powder, and purple sweet potato anthocyanin of the invention can be by this area Any prior art prepares.In the present invention, anthocyanidin is a kind of pure natural water-soluble flavonoids, and color and luster is fresh Gorgeous naturally, safe and non-toxic have no side effect, its antioxygenic property is ascorbic 20 times, is described as after water, protein, carbon aquation The seventh-largest essential nutrients after compound, fat, vitamin, mineral matter.Purple potato is a kind of very high sweet potato of anthocyanidin content Kind, there is good nutritive value and edibility, long-term consumption is with decompression, anticancer, prevention of arterial hardening and other effects.It is purple Potato anthocyanidin has very strong antioxidation activity, stability and security, and domestic and foreign scholars are carried out to its Extraction of Pigment Substantial amounts of research, is extracted using acid flux material method mostly at present, such as using citric acid-ethanol extraction, ultrasonic assistant extraction, second The methods of acid extraction.
In the present invention, the compound stabilizer formed using sodium carboxymethylcellulose with fucoidan can obtain Comparatively ideal stablizing effect.
The preparation method of drinks of the present invention is:1. prepare water chestnut bark extract, Phyllanthus embical fruit extract, purple potato cyanine Element, dandelion extract, fucoidan isoreactivity composition;2. by each component mixing preparation in proportion;3. it will adjust Beverage after matching somebody with somebody is deaerated using water circulating pump, under 0.09 Mpa vacuums, is deaerated 5 minutes;4. by the beverage filling after degassing Into vial, then cover;5. filling beverage bottle is put into 95 DEG C -100 DEG C of hot water, after sterilizing 30 min, take out, Room temperature is cooled under field conditions (factors).
Particularly, the reagent employed in the present invention is food-grade.
The present invention passes through water chestnut bark extract, Phyllanthus embical fruit extract, purple sweet potato anthocyanin, dandelion extract, sea-tangle brown alga Synergistic compounding between polysaccharide sulfate component, the anti-inflammatory for reaching good is antibacterial and immunoregulation effect, the antibacterial work of its anti-inflammatory With mainly being reached by one of approach of following several respects:Suppress migration of the leucocyte to inflammation part;Activated macrophage produces Raw nitric oxide, nitric oxide is the medium molecule of immune response and inflammatory reaction, participates in body inflammatory reaction and immunological regulation, The physiology and pathologic process of the multiple systems of body are participated in, suppresses the growth of pathogenic microorganism or kills intracellular pathogen;Suppress The permeability of capillary, reduce diffusate volume;IL-6 generation can be suppressed, and lower NF- κ B nuclear translocation, suppress Th2 The cell factors such as IL-4, IL-5, IL-13 are produced, suppresses the release of Pro-inflammatory mediator, adjusts some inflammation-related factors Generate and play antiinflammatory action;Strengthen the phagocytic activity of macrophage.
Macrophage is the important immunocyte of body, is all risen in body defenses infection, homeostasis and immunosurveillance Important effect, can be stimulated and activate, the macrophage after being activated by bacteriotoxin, toxic effect molecule, cell factor etc. Cell can produce many bioactive molecules, such as nitric oxide(Nitric oxide, NO), interleukins (Interleukin, IL), TNF(Tumor necrosis factor, TNF)Deng, many of which all with it is immune should Answer and inflammation-related.
NF- κ B be regulate and control immune response one group of pleiotropic transcription factor, its mainly play regulating cell propagation with apoptosis, The effect of immunization inflammatory reaction, play an important roll in the activation of macrophage.It can be participated in spite of a plurality of signal pathway The activation of macrophage, but almost all of signal finally can all be attributed to macrophage produce the related cell factor of inflammation and Nitric oxide synthase type(Inducible nitric oxide synthase, iNOS)Expression.
Cause macrophage activation to pass through specific receptor to mediate, the specific receptor be referred to as pattern-recognition by Body, extrinsic ligand can be identified in the starting stage of immune response.Macrophage to the present invention active component identification it is related by Body mainly has Toll-like receptor(Tolllike receptors, TLRs), LPS receptor CD14(cluster of differentiation 14), C3-receptor(Complement receptor 3, CR3), scavenger receptor(scavenger Receptor, SR), mannose receptor(Mannose receptor, MR)And BMDC correlation c-type agglutinin 1 (Dendritic cell associateded C-type lectin-1, Dectin-1)Deng.In addition, the present invention activity into The activation that endocytosis mediates macrophage can also be passed through by dividing.Occupied an leading position by receptor-mediated macrophage activation process. Participate in the activation of macrophage in spite of a plurality of signal pathway, but each approach can directly or indirectly induced NF-KB activation, And then the generation for causing related gene to be transcribed, play immunoregulation effect by lowering NF- κ B activity.
The beneficial effects of the present invention are:
Water chestnut skin is a kind of fruits and vegetables resource with local characteristic.Its unique medical value and abundant active skull cap components contain Amount, it is widely applied and provides the foundation for it.But it is recognized current consumer and acceptance level is also less high, especially in China The north.Therefore, the present invention uses water chestnut skin as primary raw material, by being compounded with other components synergistic, develops a kind of functional form Beverage products, water chestnut skin is sufficiently utilized.
The present invention is mutually mixed using different types of plant component, is learnt from other's strong points to offset one's weaknesses, and can be made into that best in quality, taste is better Mixed plant type beverage.Solve the problems, such as that the fruit juice of single taste in the market has been difficult to meet the needs of people, both It can meet that people for the pursuit in mouthfeel, also can further meet the pursuit on health diet.
Embodiment
Technical scheme is clearly and completely described below in conjunction with embodiment, it is clear that described reality It is only part of the embodiment of the present invention to apply example, rather than whole embodiments.
Embodiment 1
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Water chestnut bark extract 110, Phyllanthus embical fruit extract 33rd, purple sweet potato anthocyanin 7, sodium carboxymethylcellulose 7, xylitol 21, citric acid 16, malic acid 22, dandelion extract 32nd, fucoidan 14, water 112.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20,0.3% pectase is added by the mass ratio of raw material, 28min is digested at 45 DEG C;Carry out destroy the enzyme treatment, Filtering, leaves and takes filtrate.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, by the original quality of raw material than adding 0.8% pectase, reach 3.5 using lemon acid for adjusting pH, 50 Digest 22min at DEG C, after enzymolysis, enzymolysis liquid is boiled into 30 min enzyme deactivations, is cooled to room temperature, filtered with the nylon cloth of 200 mesh, then Filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane, obtains the Phyllanthus embical fruit extract solution of clarification.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25 are fed intake, and 100 DEG C are returned in water-bath Stream 2 h of extraction, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion extraction Thing dry powder.
Embodiment 2
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Include the raw material of following composition by weight:Water chestnut skin carries Take thing 105, Phyllanthus embical fruit extract 28, purple sweet potato anthocyanin 6, sodium carboxymethylcellulose 6, xylitol 19, citric acid 14, apple Tartaric acid 19, dandelion extract 27, fucoidan 11, water 110.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20,0.1% pectase is added by the mass ratio of raw material, 10min is digested at 45 DEG C;Carry out destroy the enzyme treatment, Filtering, leaves and takes filtrate.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, add 0.5%% pectase by the original quality ratio of raw material, reach 3.5 using lemon acid for adjusting pH, Digest 15min at 50 DEG C, after enzymolysis, enzymolysis liquid boiled into 30 min enzyme deactivations, is cooled to room temperature, filtered with the nylon cloth of 200 mesh, Filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane again, obtains the Phyllanthus embical fruit extract solution of clarification.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25 are fed intake, and 100 DEG C are returned in water-bath Stream 2 h of extraction, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion extraction Thing dry powder.
Embodiment 3
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Water chestnut bark extract 115, Phyllanthus embical fruit extract 35th, purple sweet potato anthocyanin 8, sodium carboxymethylcellulose 8, xylitol 22, citric acid 17, malic acid 26, dandelion extract 34th, fucoidan 15, water 115.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20,0.6% pectase is added by the mass ratio of raw material, 60min is digested at 45 DEG C;Carry out destroy the enzyme treatment, Filtering, leaves and takes filtrate.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, by the original quality of raw material than adding 1.2% pectase, reach 3.5 using lemon acid for adjusting pH, 50 Digest 35min at DEG C, after enzymolysis, enzymolysis liquid is boiled into 30 min enzyme deactivations, is cooled to room temperature, filtered with the nylon cloth of 200 mesh, then Filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane, obtains the Phyllanthus embical fruit extract solution of clarification.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25 are fed intake, and 100 DEG C are returned in water-bath Stream 2 h of extraction, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion extraction Thing dry powder.
Embodiment 4
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Water chestnut bark extract 97, Phyllanthus embical fruit extract 23rd, purple sweet potato anthocyanin 5, sodium carboxymethylcellulose 5, xylitol 16, citric acid 13, malic acid 16, dandelion extract 22nd, fucoidan 9, water 100.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20,0.2% pectase is added by the mass ratio of raw material, 18min is digested at 45 DEG C;Carry out destroy the enzyme treatment, Filtering, leaves and takes filtrate.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, by the original quality of raw material than adding 0.7% pectase, reach 3.5 using lemon acid for adjusting pH, 50 Digest 18min at DEG C, after enzymolysis, enzymolysis liquid is boiled into 30 min enzyme deactivations, is cooled to room temperature, filtered with the nylon cloth of 200 mesh, then Filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane, obtains the Phyllanthus embical fruit extract solution of clarification.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25 are fed intake, and 100 DEG C are returned in water-bath Stream 2 h of extraction, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion extraction Thing dry powder.
Embodiment 5
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Water chestnut bark extract 122, Phyllanthus embical fruit extract 41st, purple sweet potato anthocyanin 10, sodium carboxymethylcellulose 9, xylitol 25, citric acid 19, malic acid 28, dandelion extract 38th, fucoidan 17, water 120.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20,0.5% pectase is added by the mass ratio of raw material, 50min is digested at 45 DEG C;Carry out destroy the enzyme treatment, Filtering, leaves and takes filtrate.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, by the original quality of raw material than adding 1.0% pectase, reach 3.5 using lemon acid for adjusting pH, 50 Digest 30min at DEG C, after enzymolysis, enzymolysis liquid is boiled into 30 min enzyme deactivations, is cooled to room temperature, filtered with the nylon cloth of 200 mesh, then Filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane, obtains the Phyllanthus embical fruit extract solution of clarification.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25 are fed intake, and 100 DEG C are returned in water-bath Stream 2 h of extraction, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion extraction Thing dry powder.
Comparative example 1
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Phyllanthus embical fruit extract 33, purple sweet potato anthocyanin 7, carboxylic Sodium carboxymethylcellulose pyce 7, xylitol 21, citric acid 16, malic acid 22, dandelion extract 32, sea-tangle algal polysaccharide sulfuric acid Ester 14, water 112.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, by the original quality of raw material than adding 0.8% pectase, reach 3.5 using lemon acid for adjusting pH, 50 Digest 22min at DEG C, after enzymolysis, enzymolysis liquid is boiled into 30 min enzyme deactivations, is cooled to room temperature, filtered with the nylon cloth of 200 mesh, then Filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane, obtains the Phyllanthus embical fruit extract solution of clarification.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25 are fed intake, and 100 DEG C are returned in water-bath Stream 2 h of extraction, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion extraction Thing dry powder.
Comparative example 2
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Water chestnut bark extract 110, purple sweet potato anthocyanin 7, Sodium carboxymethylcellulose 7, xylitol 21, citric acid 16, malic acid 22, dandelion extract 32, sea-tangle algal polysaccharide sulphur Acid esters 14, water 112.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20,0.3% pectase is added by the mass ratio of raw material, 28min is digested at 45 DEG C;Carry out destroy the enzyme treatment, Filtering, leaves and takes filtrate.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25 are fed intake, and 100 DEG C are returned in water-bath Stream 2 h of extraction, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion extraction Thing dry powder.
Comparative example 3
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Water chestnut bark extract 110, Phyllanthus embical fruit extract 33rd, purple sweet potato anthocyanin 7, sodium carboxymethylcellulose 7, xylitol 21, citric acid 16, malic acid 22, sea-tangle algal polysaccharide sulphur Acid esters 14, water 112.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20,0.3% pectase is added by the mass ratio of raw material, 28min is digested at 45 DEG C;Carry out destroy the enzyme treatment, Filtering, leaves and takes filtrate.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, by the original quality of raw material than adding 0.8% pectase, reach 3.5 using lemon acid for adjusting pH, 50 Digest 22min at DEG C, after enzymolysis, enzymolysis liquid is boiled into 30 min enzyme deactivations, is cooled to room temperature, filtered with the nylon cloth of 200 mesh, then Filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane, obtains the Phyllanthus embical fruit extract solution of clarification.
Comparative example 4
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Water chestnut bark extract 110, Phyllanthus embical fruit extract 33rd, sodium carboxymethylcellulose 7, xylitol 21, citric acid 16, malic acid 22, dandelion extract 32, sea-tangle brown alga are more Sugar sulfate 14, water 112.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20,0.3% pectase is added by the mass ratio of raw material, 28min is digested at 45 DEG C;Carry out destroy the enzyme treatment, Filtering, leaves and takes filtrate.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, by the original quality of raw material than adding 0.8% pectase, reach 3.5 using lemon acid for adjusting pH, 50 Digest 22min at DEG C, after enzymolysis, enzymolysis liquid is boiled into 30 min enzyme deactivations, is cooled to room temperature, filtered with the nylon cloth of 200 mesh, then Filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane, obtains the Phyllanthus embical fruit extract solution of clarification.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25 are fed intake, and 100 DEG C are returned in water-bath Stream 2 h of extraction, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion extraction Thing dry powder.
Comparative example 5
A kind of water chestnut skin drinks, include the raw material of following composition by weight:Water chestnut bark extract 110, Phyllanthus embical fruit extract 33rd, purple sweet potato anthocyanin 7, sodium carboxymethylcellulose 7, xylitol 21, citric acid 16, malic acid 22, dandelion extract 32nd, water 112.
Further, the preparation method of the water chestnut bark extract is:By fresh horseshoe skin wash clean, dry, crush; Material-water ratio m:V=1:20,0.3% pectase is added by the mass ratio of raw material, 28min is digested at 45 DEG C;Carry out destroy the enzyme treatment, Filtering, leaves and takes filtrate.
Further, the preparation method of the Phyllanthus embical fruit extract is:By Phyllanthus embical fruit and pure water according to m:V=1:20 Ratio is mixed, and after boiling 30 min, the Phyllanthus embical fruit and aqueous mixtures that will be cooled to room temperature are added in colloid mill, and regulation gap is 0.1 mm, grind three times, by the original quality of raw material than adding 0.8% pectase, reach 3.5 using lemon acid for adjusting pH, 50 Digest 22min at DEG C, after enzymolysis, enzymolysis liquid is boiled into 30 min enzyme deactivations, is cooled to room temperature, filtered with the nylon cloth of 200 mesh, then Filtrate is filtered by molecular cut off for 2 kDa device for ultrafiltration membrane, obtains the Phyllanthus embical fruit extract solution of clarification.
Further, the preparation method of the dandelion extract is:Dry dandelion herb is taken, is crushed to 40 mesh, by nothing Water-ethanol is formulated as 70% volumetric concentration, as digestion agent, according to solid-liquid ratio m:V is 1:25 are fed intake, and 100 DEG C are returned in water-bath Stream 2 h of extraction, filtering and concentrating is carried out after terminating, and the leaching liquor that concentration is 1 g/mL is made, and freeze-drying obtains dandelion extraction Thing dry powder.
Measure of merit
Method of testing in the present invention is reference《Health food is examined and assessment technique enforcement of regulations handbook》, according to the state of correlation Family standard formulation, the present invention in all experimental datas processing using the softwares of SPSS 16.0 carry out statistical analysis.
Beverage sensory evaluation
Using sensory evaluation method, the color and luster, flavor, mouthfeel of water chestnut skin flavor beverage are scored for deliberated index, ask 8 tools There is senior beverage to judge the expert of experience to taste, given a mark after control sensory judgments table identification, full marks 100 divide, and remove a highest Point and one minimum point, then average as evaluation result.Standards of grading such as following table.
Tested by sensory evaluation, the sensory evaluation score such as following table of each embodiment and comparative example.
2. antiphlogistic effects are tested
Antiphlogistic effects test is carried out to embodiment 1-5 and comparative example 1-5, mouse 72 is screened, is bisected into 12 groups at random.Setting one Group blank control group, gives 0.2mL/20g physiological saline;One positive drug control group, 1 property gives 0.2mL/20g hydrogenations can Loose parenteral solution.Embodiment 1-5 and comparative example 1-5 is test group, dosage 0.2mL/20g, successive administration 10d.Last is given Medicine 1h, mouse right ear is wide to apply caused by dimethylbenzene xylene inflammation.Mouse is put to death after 1h, cuts ears diameter 9mm card punch in same portion Auricle is laid in position, weighs and records.The experimental results are shown inthe following table.
Note:" a " represents there is significant difference with blank group;" b " represents there is significant difference with positive group.
Antiphlogistic effects confirmatory experiment
2.1 mice caused by dimethylbenzene xylene auricle edemas are tested
Mouse 50, is randomly divided into 5 groups.A groups are blank control group, give 0.2mL/20g physiological saline, and B, C, D group are experiment Group, embodiment 1 solution 0.1mL/20g, 0.2mL/20g and 0.4mL/20g, successive administration 10d are given respectively.E groups are positive drug Thing control group, 0.2mL/20g hydrocortisone parenteral solutions are disposably given, it is as a result as shown in the table.
Note:" a " represents there is significant difference with blank group;" b " represents there is significant difference with positive group.
2.2 swollen hyperplasia of rat granuloma are tested
Rat 25 is taken, with 1% anaesthetized with pentobarbital, lower abdomen unhairing sterilization, by 50mg sterile cotton balls implantation left side groin Subcutaneously, sew up a wound.It is postoperative to be randomly divided into 5 groups, every group 5, start to be administered after 1h, be grouped and be administered same 2.1, successive administration 10d.Take out granuloma induced by implantation of cotton pellets tissue within 11st day, 60 DEG C of drying, weigh and record in drying baker, as a result as shown in the table.
Note:" a " represents there is significant difference with blank group;" b " represents there is significant difference with positive group.
3. immunomodulatory effect is tested
To be dried by drinks thickening made from embodiment 1, be prepared into it is powdered, as test sample.It is positive right According to product:Its vigorous and graceful board Soybean Peptide albumen powder;Source:Beijing Tongrentang Health Pharmaceutical Co., Ltd.;Adult is clinical to be recommended to use Amount:0.167g.kg-1.d-1
Animal subject strain and rank:BALB/c mouse, Hartley cavys, SPF levels;Quantity and sex:BALB/c is small Mouse 60, male;Hartley cavys 6,.Buy body weight BALB/c mouse 17-19g, Hartley cavy 250- 350g.During experiment, mouse ad lib drinking-water, mouse state is observed daily.
Dose design is carried out according to the basic condition of sample, sample it is basic, normal, high by adult 5 times of quantity, 10 times, 20 times of design dosage, respectively 0.15g.kg-1.d-1、 0.30g.kg-1.d-1 、0.60g.kg-1.d-1;Positive group dosage is proportionately 5 times of designs of people's recommended dose, are 0.835g.kg-1.d-1.60 male mices are respectively divided into 5 groups at random(Blank control group, The basic, normal, high dosage group of positive controls, given the test agent), every group 12, wherein 2 are only used as standby animal.Each group mouse is daily By 0.1mL/10g body weight gavage respective sample liquid, blank group gives equivalent pure water, 1 time a day, successive administration 30 days.Experiment Beginning, off-test are weighed in 1 time, are weighed in weekly during experiment 1 time.
3.1 mouse lymphocyte transformation experiments
Last dose 1h, it is sterile to take spleen to be placed in the culture dish for filling RPMI-1640 nutrient solutions, and 200 eye mesh screens are placed, with note Emitter piston is ground above it is made splenocyte suspension, packing splenocyte suspension to the centrifuge tube added with lymphocyte separation medium In.400g, 20min is centrifuged at 20 DEG C, centrifugation finishes, and takes out centrifuge tube medium size lymphocyte layer centrifuge tube, adds RPMI-1640 trainings Nutrient solution is washed for several times, 250g, and 10min is centrifuged at 4 DEG C.After washing, abandoning supernatant adds complete medium, and piping and druming is equal It is even, expect that blue dyeing counting ensures more than 95% living cell rate with platform, cell concentration is adjusted to 2.5 × 105Individual/ml.
Holes is divided to add in 48 well culture plates every part of splenocyte suspension, per hole 0.5ml, a hole adds 25 ul ConA liquid (Equivalent to 5ug/ml), another hole is placed in 5%CO as control2,37℃ CO2Gently sucked per hole after 48h is cultivated in incubator The ml of clear liquid 0.3,0.3 ml RPMI-1640 nutrient solutions are added, while add the ul/ holes of 8 reagents of CCK 50, continue to cultivate 2 h. After terminating culture, piping and druming is uniform, is then dispensed into 96 well culture plates, makees 3 parallel laboratory tests per hole, and 450 nm are surveyed with ELIASA OD value under wavelength.The multiplication capacity of lymphocyte is by adding the OD value in ConA holes and being not added with the optical density in ConA holes The difference of value represents.The experimental results are shown inthe following table.
Note:" a " represents there is significant difference with blank group;" b " represents there is significant difference with positive group.
3.2 mouse humoral immune functions(Serum hemolysin)Influence
Administration 25 days, taken off the sheep blood 3 times of fiber with brine (2000r/min, 10min), in hematocrit SRBC plus Enter physiological saline and be made into 2%(v/v)Cell suspension, every mouse peritoneal injection 0.2ml, at the 30th day, after 1h is administered, weigh, All mouse extract eyeball and take blood 0.8ml to stand 1h in centrifuge tube, centrifuge 2000r/min, 10min, collect serum.Cervical vertebra takes off Mortar puts to death mouse.300 times of serum is diluted with SA buffer solutions takes 1ml to put in test tube, adds 10% again(v/v) SRBC0.5ml, with SA buffer solutions 9 times of complement 1ml is diluted, control tube is set(Serum is replaced with SA), 37 DEG C of waters bath with thermostatic control 20min, then ice bath, it is then centrifuged for 2000r/min, 10min.Supernatant 1ml is taken in new test tube, add 3.75ml Dou Shi reagents, 0.25ml 10%(v/v)SRBC, fully mix, stand 10min, using control tube as blank, determine each Guan Guangmi at 540nm respectively Angle value, the amount of hemolysin is with half hemolytic value(HC50)Represent.As a result it is as shown in the table.The experimental results are shown inthe following table.
Note:" a " represents there is significant difference with blank group;" b " represents there is significant difference with positive group.
The measure that 3.3 mouse monokaryons-macrophage phagocytic function and immune organ influence
Last dose 1h, by body weight from tail vein injection physiological saline 1:The india ink of 3 dilutions(10mL/kg), treat prepared Chinese ink Injection, immediately timing.2,10min after injection prepared Chinese ink, respectively from orbital venous plexus 20 μ L of blood sampling, and 2mL 0.1% is added to immediately Na2CO3In solution, with Na2CO3Solution makees blank control, and OD value is determined at 600nm wavelength with spectrophotometer(OD), Calculate phagocytic index a.
Dislocate and put to death after mouse blood sampling, solution takes liver, spleen and thymus gland, accurate after filter paper blots internal organs peripheral blood Weigh, calculate internal organs/body weight ratio.The experimental results are shown inthe following table.
Note:" a " represents there is significant difference with blank group;" b " represents there is significant difference with positive group.
3.4 NK cytoactive detections
Last dose 1h, it is sterile to take spleen, it is sterile to take spleen to be placed in the culture dish for filling RPMI-1640 nutrient solutions, and place 200 mesh Screen cloth, ground above it with syringe piston and splenocyte suspension is made, packing splenocyte suspension is to added with separation of lymphocytes In the centrifuge tube of liquid.400g, 20min is centrifuged at 20 DEG C, centrifugation finishes, and takes out centrifuge tube medium size lymphocyte layer centrifuge tube, adds RPMI-1640 nutrient solutions are washed for several times, 250g, and 10min is centrifuged at 4 DEG C.After washing, abandoning supernatant, training completely is added Base is supported, piping and druming is uniform, expects blue dyeing counting viable count with platform(Should be more than 95%), finally with the complete culture solutions of RPMI 1640 It is 2 × 10 to adjust cell concentration7Individual/mL.
In proportion by effector cell and target cell(50:1)Each 100ul adds U-shaped 96 orifice plate;Target cell and nutrient solution are each 100ul adds target cell Spontaneous release hole;Target cell and each 100ul of 0.25%Triton add target cell maximum release aperture.Every group It is all provided with 3 parallel holes, 37 DEG C, after 5%CO2 incubator cultures 4h, centrifuges 1500rpm, 5min.Per hole Aspirate supernatant 100ul in Flat 96 orifice plate, LDH matrix liquid 100ul/ holes are added, react 3-10min, add 1mol/ml HCL 30ul/ holes, enzyme mark 490nm determines OD value.The experimental results are shown inthe following table.
Note:" a " represents there is significant difference with blank group;" b " represents there is significant difference with positive group.
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, and do not carrying on the back In the case of spirit or essential attributes from the present invention, the present invention can be realized in other specific forms.Therefore, no matter from which From the point of view of a bit, embodiment all should be regarded as exemplary, and be nonrestrictive, the scope of the present invention will by appended right Ask rather than described above limits, it is intended that all changes in the implication and scope of the equivalency of claim will be fallen Include in the present invention.
Moreover, it will be appreciated that although the present specification is described in terms of embodiments, not each embodiment is only wrapped Containing an independent technical scheme, this narrating mode of specification is only that those skilled in the art should for clarity Using specification as an entirety, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art It is appreciated that other embodiment.It is noted that the technical characteristic not being described in detail in the present invention, can pass through this Field any prior art is realized.

Claims (6)

1. a kind of water chestnut skin drinks, it is characterised in that include the raw material of following composition by weight:Water chestnut bark extract 97- 122nd, Phyllanthus embical fruit extract 23-41, purple sweet potato anthocyanin 5-10, sodium carboxymethylcellulose 5-9, xylitol 16-25, citric acid 13-19, malic acid 16-28, dandelion extract 22-38, fucoidan 9-17, water 100-120.
2. water chestnut skin drinks according to claim 1, it is characterised in that include the raw material of following composition by weight:Water chestnut Shepherd's purse bark extract 105-115, Phyllanthus embical fruit extract 28-35, purple sweet potato anthocyanin 6-8, sodium carboxymethylcellulose 6-8, xylose Alcohol 19-22, citric acid 14-17, malic acid 19-26, dandelion extract 27-34, fucoidan 11- 15th, water 110-115.
3. water chestnut skin drinks according to claim 1, it is characterised in that include the raw material of following composition by weight:Water chestnut Shepherd's purse bark extract 110, Phyllanthus embical fruit extract 33, purple sweet potato anthocyanin 7, sodium carboxymethylcellulose 7, xylitol 21, citric acid 16th, malic acid 22, dandelion extract 32, fucoidan 14, water 112.
4. water chestnut skin drinks according to claim 1, it is characterised in that the preparation method of the water chestnut bark extract For:By fresh horseshoe skin wash clean, dry, crush;In material-water ratio m:V=1:20, add 0.1%-0.6% by the mass ratio of raw material Pectase, digest 10-60min at 45 DEG C;Destroy the enzyme treatment is carried out, filtering, leaves and takes filtrate.
5. water chestnut skin drinks according to claim 1, it is characterised in that the preparation method of the Phyllanthus embical fruit extract For:By Phyllanthus embical fruit and pure water according to m:V=1:20 ratio mixing, after boiling 30 min, will be cooled to room temperature Phyllanthus embical fruit and Aqueous mixtures are added in colloid mill, and regulation gap is 0.1 mm, are ground three times, by the original quality of raw material than adding 0.5%- 1.2% pectase, reach 3.5 using lemon acid for adjusting pH, digest 15-35min at 50 DEG C, after enzymolysis, enzymolysis liquid is boiled 30 Min enzyme deactivations, are cooled to room temperature, are filtered with the nylon cloth of 200 mesh, then filtrate is set by molecular cut off for 2 kDa milipore filters Standby filtering, obtain the Phyllanthus embical fruit extract solution of clarification.
6. water chestnut skin drinks according to claim 1, it is characterised in that the preparation method of the dandelion extract For:Dry dandelion herb is taken, is crushed to 40 mesh, absolute ethyl alcohol is formulated as 70% volumetric concentration, as digestion agent, according to feed liquid Compare m:V is 1:25 feed intake, and 100 DEG C of h of refluxing extraction 2 in water-bath, filtering and concentrating are carried out after terminating, and it is 1 g/ that concentration, which is made, ML leaching liquor, freeze-drying obtain dandelion extract dry powder.
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Cited By (5)

* Cited by examiner, † Cited by third party
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CN108813270A (en) * 2018-04-25 2018-11-16 惠州嘉联生物科技开发有限公司 A kind of composite enzyme of strengthen immunity and preparation method thereof
CN108936136A (en) * 2018-04-25 2018-12-07 许昌学院 A kind of cool to reduce pathogenic fire beverage and preparation method thereof
CN111000107A (en) * 2019-12-30 2020-04-14 贺州学院 Preparation method of water chestnut flavor peel powder, and solid beverage and hard candy prepared from water chestnut flavor peel powder
CN117322629A (en) * 2023-11-23 2024-01-02 韶关学院 Preparation method of horseshoe lipid-lowering composition based on fermentation enzymolysis technology
CN117322629B (en) * 2023-11-23 2024-05-17 韶关学院 Preparation method of horseshoe lipid-lowering composition based on fermentation enzymolysis technology

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