CN108434171A - A kind of arrowhead polysaccharide composition and preparation method thereof preventing chemical damage - Google Patents
A kind of arrowhead polysaccharide composition and preparation method thereof preventing chemical damage Download PDFInfo
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- CN108434171A CN108434171A CN201810100520.8A CN201810100520A CN108434171A CN 108434171 A CN108434171 A CN 108434171A CN 201810100520 A CN201810100520 A CN 201810100520A CN 108434171 A CN108434171 A CN 108434171A
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- arrowhead
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- chemical damage
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- 239000000203 mixture Substances 0.000 title claims abstract description 33
- 150000004676 glycans Chemical class 0.000 title claims abstract description 29
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 28
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 239000000126 substance Substances 0.000 title claims abstract description 21
- 230000006378 damage Effects 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 11
- 229920002472 Starch Polymers 0.000 claims abstract description 7
- 235000019698 starch Nutrition 0.000 claims abstract description 7
- 239000008107 starch Substances 0.000 claims abstract description 7
- 229920001353 Dextrin Polymers 0.000 claims abstract description 6
- 239000004375 Dextrin Substances 0.000 claims abstract description 6
- 235000013871 bee wax Nutrition 0.000 claims abstract description 6
- 239000012166 beeswax Substances 0.000 claims abstract description 6
- 235000019425 dextrin Nutrition 0.000 claims abstract description 6
- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical compound CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 claims abstract description 6
- 230000008569 process Effects 0.000 claims abstract description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 21
- 235000019441 ethanol Nutrition 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 238000001556 precipitation Methods 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 8
- 239000012141 concentrate Substances 0.000 claims description 7
- 239000000284 extract Substances 0.000 claims description 7
- 238000005119 centrifugation Methods 0.000 claims description 6
- 230000006835 compression Effects 0.000 claims description 6
- 238000007906 compression Methods 0.000 claims description 6
- 238000010168 coupling process Methods 0.000 claims description 6
- 238000005859 coupling reaction Methods 0.000 claims description 6
- 239000012530 fluid Substances 0.000 claims description 6
- 238000005360 mashing Methods 0.000 claims description 6
- 238000003809 water extraction Methods 0.000 claims description 6
- 229920002678 cellulose Polymers 0.000 claims description 3
- 235000010980 cellulose Nutrition 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims description 3
- 239000008367 deionised water Substances 0.000 claims description 3
- 229910021641 deionized water Inorganic materials 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 239000012535 impurity Substances 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 239000003480 eluent Substances 0.000 claims description 2
- 239000002994 raw material Substances 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 239000002775 capsule Substances 0.000 abstract description 6
- 239000004615 ingredient Substances 0.000 abstract description 6
- 239000003814 drug Substances 0.000 abstract description 5
- 239000008187 granular material Substances 0.000 abstract description 5
- 239000003826 tablet Substances 0.000 abstract description 5
- 229940079593 drug Drugs 0.000 abstract description 4
- 230000003321 amplification Effects 0.000 abstract description 2
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 2
- 239000002417 nutraceutical Substances 0.000 abstract description 2
- 235000021436 nutraceutical agent Nutrition 0.000 abstract description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N carbon tetrachloride Substances ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 18
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 230000002265 prevention Effects 0.000 description 8
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 6
- 206010019837 Hepatocellular injury Diseases 0.000 description 5
- 231100000437 hepatocellular injury Toxicity 0.000 description 5
- 231100000753 hepatic injury Toxicity 0.000 description 4
- 239000004480 active ingredient Substances 0.000 description 3
- 229910052793 cadmium Inorganic materials 0.000 description 3
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 230000000050 nutritive effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000005180 public health Effects 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 240000009132 Sagittaria sagittifolia Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 229960003350 isoniazid Drugs 0.000 description 2
- QRXWMOHMRWLFEY-UHFFFAOYSA-N isoniazide Chemical compound NNC(=O)C1=CC=NC=C1 QRXWMOHMRWLFEY-UHFFFAOYSA-N 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 2
- 229960001225 rifampicin Drugs 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010067125 Liver injury Diseases 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 235000006466 Sagittaria sagittifolia Nutrition 0.000 description 1
- 244000018694 Sagittaria sinensis Species 0.000 description 1
- 235000015909 Sagittaria sinensis Nutrition 0.000 description 1
- 240000001085 Trapa natans Species 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000006286 aqueous extract Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 239000012888 bovine serum Substances 0.000 description 1
- 229940044194 cadmium Drugs 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 231100000012 chronic liver injury Toxicity 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 231100000234 hepatic damage Toxicity 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 230000008818 liver damage Effects 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000003617 peroxidasic effect Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/884—Alismataceae (Water-plantain family)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Materials Engineering (AREA)
- Biochemistry (AREA)
- Sustainable Development (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The present invention discloses a kind of arrowhead polysaccharide composition and preparation method thereof preventing chemical damage, is related to medicine and technical field of agricultural product process.The composition is made of arrowhead polysaccharide and adjunct ingredient, wherein arrowhead mass fraction of polysaccharide 10 50, the mass fraction of auxiliary element is respectively dextrin 5 10, starch 5 20, beeswax 15, dodecyl sulphate 15, it is formulated according to aforementioned proportion, and the oral preparations such as pulvis, capsule, tablet and granule is made as needed.The present invention specifies that arrowhead prevents the active component and ingredient of chemical damage by biological back tracking method, precisely develops the drug for preventing chemical damage for arrowhead resource or nutraceutical product provides new resources.The present invention provides the preparation method for the arrowhead polysaccharide composition for preventing chemical damage, based on the arrowhead polysaccharide of definite functions, addition partial supplementary material is prepared the composition, operating process is simple, is easy to industrialization amplification.
Description
Technical field
The invention belongs to technical field of agricultural product process, and in particular to a kind of arrowhead polysaccharide group for preventing chemical damage
Close object and preparation method thereof.
Background technology
Arrowhead (Sagittaria sagittifolia Linm) also known as arrowhead, arrowhead are that Alismataceae Sagittaria is perennial
Perennial root shallow water herbaceous plant.China is the source area of arrowhead, based on the southern region of rivers and lakes, and it is mostly in the majority to cultivate, especially with Jiangsu
Province Yangzhou Baoying County's arrowhead cultivated area is big, and quality is good, is known as the good reputation in " township of Chinese arrowhead ", and as national geography mark
Will product.The edible bulb of arrowhead be rich in starch, protein and multivitamin, dietary fiber, alkaloid, polyphenol and iron,
The trace element of a variety of needed by human body such as calcium, zinc, average nutritive value are above the aquatic vegetables such as lotus rhizome, water chestnut bletilla water chestnut
Average nutritive value, it is considered to be the first-class treasure in nuisanceless green health food.Arrowhead is the quite high U.S. of nutritive value
Taste delicacies.Modern pharmacology research shows that arrowhead has the multiple efficacies such as anti-oxidant, removing toxic substances, hypoglycemic, antitumor, while right
The hepatic injury that cadmium, isoniazid, rifampin induce has protective effect.
The reports such as Wu little Nan give arrowhead juice in advance can reduce cadmium and CCl4Cause acute and chronic hepatic injury (Chinese public health,
2002,18:1312-1313；Chinese public health, 1999,15:16-18).Lu Zufu and Wu little Nan etc. reports arrowhead juice to cadmium
Cause rats'liver peroxidatic reaction of lipid that there is antagonism (Chinese public health journal 1999,18:348-350；China is public to be defended
Raw 2002,18：388-389；Medical University Of Fujian's journal, 2001,35：116-118).Liao Yan etc. reports arrowhead juice to isoniazid
With rifampin share cause Liver Damage in Rats have protective effect (Beijing University of Chinese Medicine's journal, 2012,35:761-764).From with
It is upper studies have shown that in arrowhead juice chemically or the active ingredient of drug hepatic injury containing prevention and treatment, but used in above-mentioned report
Arrowhead juice is after air-drying arrowhead, by weight 1:1 adds distilled water to be ground in fiberizer, arrowhead juice obtained after press filtration,
The arrowhead Aqueous extracts that actually cold water extracts in short-term contain a certain amount of nutritional ingredient and active matter in this arrowhead liquid
Matter also contains the active ingredient for having prevention or therapeutic effect to hepatic injury, but existing literature does not continue to track arrowhead with pre-
The active ingredient of anti-chemical hepatic injury effect.The present invention is determined by biological back tracking method prevents chemical damage in arrowhead
Active component and ingredient, and be prepared into composition and be used.
Invention content
The object of the present invention is to provide a kind of arrowhead polysaccharide compositions and preparation method thereof preventing chemical damage, are
Prevent chemical damage and new drug resource is provided, while also solving effective portion that arrowhead prevents or treats chemical damage
Position and the indefinite problem of ingredient.
A kind of arrowhead polysaccharide composition preventing chemical damage, is made of the raw material of following masses number：
A kind of preparation method of above-mentioned arrowhead polysaccharide composition preventing chemical damage, carries out as steps described below：
(1) preparation of arrowhead polysaccharide
1. arrowhead pre-processes：It is spare after the removal of impurities of fresh arrowhead, cleaning, slice.
2. being beaten：Mass volume ratio 1 is added after slice immediately:10(g:ML water mashing), mashing liquid cross 60 mesh industry filter
Cloth.
3. Subcritical Water Extraction：Filtered fluid is injected into subcritical abstraction kettle, control magnetic-coupling stirrer 300- through compression pump
600 turns/min, extracting pressure 1-12MPa, 100-180 DEG C of temperature extracts 10-30min, after, supernatant is collected in centrifugation,
As arrowhead Subcritical Water Extraction liquid.
4. concentrating：It will be concentrated into the 1/2 of original volume by the arrowhead liquid 3. obtained.
5. alcohol precipitation：95% ethyl alcohol of its 4 times of volumes is added to concentrate, after standing for 24 hours, centrifuges precipitation.
6. enrichment/separation：Will after precipitation redissolves, on the DEAE-52 celluloses that have balanced, be to elute with deionized water
Liquid collects the part of washing, pressurized to be concentrated to give concentrate.
7. drying：After concentrate is dried, as arrowhead polysaccharide.
(2) preparation of composition
The mass fraction of arrowhead mass fraction of polysaccharide 10-50 in composition, auxiliary element are respectively dextrin 5-10, starch 5-
20, beeswax 1-5, dodecyl sulphate 1-5, are formulated according to aforementioned proportion, and pulvis, capsule, tablet are made as needed
With the oral preparations such as granule.
Beneficial effects of the present invention：
(1) present invention specifies that arrowhead prevents the active component and ingredient of chemical damage by biological back tracking method, is
Arrowhead resource precisely develops the drug for preventing chemical damage or nutraceutical product provides new resources.
(2) the present invention provides prevent chemical damage arrowhead polysaccharide composition preparation method, the composition with
Based on the arrowhead polysaccharide of definite functions, addition partial supplementary material is prepared, operating process is simple, is easy to industrialization amplification.
(3) composition powder produced by the present invention for preventing chemical damage, can be directly added into medically acceptable
Auxiliary material or auxiliary element pulvis, granule, tablet, capsule etc. oral preparation is made, different crowd demand can be met.
Specific implementation mode
In order to make the foregoing objectives, features and advantages of the present invention clearer and more comprehensible, below by the specific of the present invention
Embodiment is described in detail.Elaborate many details in the following description to facilitate a thorough understanding of the present invention, but
It is that the present invention can also be implemented using other different from other manner described here, those skilled in the art can not disobey
Similar popularization is done in the case of back of the body intension of the present invention, therefore the present invention is not limited by following public specific embodiment.
This patent arrowhead polysaccharide composition is to CCl4Or alcohol causes the prevention effect of hepatocellular injury to use human liver cancer thin
Born of the same parents HepG2 is cell model, which is purchased from Beijing Chinese Academy of Sciences's biochemistry and Institute of Cell Biology, the training of HepG2 cells
It supports in containing 10% inactivation newborn bovine serum DMEM culture solutions, is placed in 37 DEG C, 5%CO2, it cultivates in the incubator of saturated humidity,
Cell adherent growth passes on 1 time per 2-3d.Experiment CCl4It is dissolved in 0.5% dimethyl sulfoxide (DMSO) (DMSO) solution, is configured to
The CCl4 solution of 24mM, alcohol are made into the alcohol liquid of final concentration 2.4% with complete culture solution.The side of composition prevention effect research
Method is as follows：
The HepG2 cells of exponential phase are pressed 1 × 104A/hole is inoculated in 96 orifice plates, in 37 DEG C, 5%CO2In incubator
It is incubated for 24 hours, in advance with after the composition of final concentration of 300 μ g/mL incubation 4h, then with final concentration 24mM CCl4Handle cell 12h
Or the ethanol solution of final concentration 2.4% handles cell 12h, after, culture solution is absorbed, is cleaned 2 times with PBS, 100 are added per hole
μ L MTT (1mg/mL) continue to cultivate 4h, remove supernatant, 150 μ L DMSO are added per hole, fully vibrates 10min, uses microplate reader
Absorbance value is measured at 490nm, calculates cell survival rate, survival rate (%)=(administration group absorbance value/control group extinction
Angle value) × 100%.
Embodiment 1
The preparation of arrowhead polysaccharide：After the removal of impurities of fresh arrowhead, cleaning, slice, mass volume ratio 1 is added immediately:10(g:
ML water mashing), mashing liquid cross 60 mesh industrial filter cloths；Filtered fluid is injected through compression pump in subcritical abstraction kettle, magnetic coupling is controlled
Blender 300 turns/min, extracting pressure 1MPa are closed, 100 DEG C of temperature extracts 10min, after, supernatant is collected in centrifugation；It will
Supernatant concentration is to the 1/2 of original volume；95% ethyl alcohol of its 4 times of volumes is added into concentrate, after standing for 24 hours, centrifuges
Precipitation；Will after precipitation redissolves, on the DEAE-52 celluloses that have balanced collect the portion of washing using deionized water as eluent
Point, drying, as arrowhead polysaccharide, yield 10.57%, wherein polyoses content 80.13%, protein content 2.54% after concentration.
The preparation of composition：The mass fraction of arrowhead mass fraction of polysaccharide 10 in composition, auxiliary element is respectively dextrin
10, starch 20, beeswax 5, dodecyl sulphate 5 are directly prepared into pulvis or capsule, or tablet and particle are made by solvent of water
The oral preparations such as agent.The composition is to CCl4The prevention effect effect of hepatocellular injury is caused to be shown in Table 1 with alcohol.
Embodiment 2
Arrowhead pre-treating method will obtain filtered fluid and inject subcritical abstraction kettle through compression pump, control magnetic coupling with embodiment 1
Blender 400 turns/min, extracting pressure 5MPa are closed, 140 DEG C of temperature extracts 20min, after, supernatant is collected in centrifugation；Its
For subsequent processing with embodiment 1, it is 17.72%, wherein polyoses content 84.31%, protein content to obtain arrowhead polysaccharide yield
1.18%.
The preparation of composition：The mass fraction of arrowhead mass fraction of polysaccharide 30 in composition, auxiliary element is respectively dextrin
8, starch 7, beeswax 3, dodecyl sulphate 3 are directly prepared into pulvis or capsule, or tablet and granule are made by solvent of water
Etc. oral preparations.The composition is to CCl4The prevention effect effect of hepatocellular injury is caused to be shown in Table 1 with alcohol.
Embodiment 3
Arrowhead pre-treatment is injected through compression pump in subcritical abstraction kettle with embodiment 1, by filtered fluid, control magnetic coupling stirring
Device 600 turns/min, extracting pressure 12MPa, 180 DEG C of temperature extract 10min, after, supernatant is collected in centrifugation；It is subsequently located
Reason is with embodiment 5, which is 18.67%, polyoses content 86.09, protein content 1.39%.
The preparation of composition：The mass fraction of arrowhead mass fraction of polysaccharide 50 in composition, auxiliary element is respectively dextrin
5, starch 5, beeswax 1, dodecyl sulphate 1 are directly prepared into pulvis or capsule, or tablet and granule are made by solvent of water
Etc. oral preparations.The composition is to CCl4The prevention effect effect of hepatocellular injury is caused to be shown in Table 1 with alcohol.
1 composition of table is to CCl4The prevention effect of the hepatocellular injury of induction
| Group | Survival rate (%) |
| Normal control | 100±2.54 |
| 0.5%DMSO | 98.72±3.87 |
| 24mM CCl4 | 34.03±0.45 |
| CCl4+ embodiment 1 | 75.42±2.46 |
| CCl4+ embodiment 2 | 83.95±5.08 |
| CCl4+ embodiment 3 | 90.49±2.45 |
| 2.4% alcohol | 63.68±3.46 |
| Alcohol+embodiment 1 | 81.06±5.10 |
| Alcohol+embodiment 2 | 85.03±3.76 |
| Alcohol+embodiment 3 | 92.18±5.69 |
Claims (3)
1. it is a kind of prevent chemical damage arrowhead polysaccharide composition, it is characterised in that its by following masses number raw material group
At：
2. a kind of preparation method of arrowhead polysaccharide composition preventing chemical damage according to claim 1, special
Sign is to carry out as steps described below：
(1) preparation of arrowhead polysaccharide
1. arrowhead pre-processes：It is spare after the removal of impurities of fresh arrowhead, cleaning, slice；
2. being beaten：Mass volume ratio 1 is added after slice immediately:10(g:ML water mashing), mashing liquid cross 60 mesh industrial filter cloths；
3. Subcritical Water Extraction：Filtered fluid is injected into subcritical abstraction kettle, control magnetic-coupling stirrer 300-600 through compression pump
Turning/min, extracting pressure 1-12MPa, 100-180 DEG C of temperature extracts 10-30min, after, centrifugation collects supernatant, as
Arrowhead Subcritical Water Extraction liquid；
4. concentrating：It will be concentrated into the 1/2 of original volume by the arrowhead liquid 3. obtained；
5. alcohol precipitation：95% ethyl alcohol of its 4 times of volumes is added to concentrate, after standing for 24 hours, centrifuges precipitation；
6. enrichment/separation：Will after precipitation redissolves, on the DEAE-52 celluloses that have balanced, using deionized water as eluent,
The part of washing is collected, it is pressurized to be concentrated to give concentrate；
7. drying：After concentrate is dried, as arrowhead polysaccharide；
(2) preparation of composition
Arrowhead mass fraction of polysaccharide 10-50 in composition, the mass fraction of auxiliary element be respectively dextrin 5-10, starch 5-20,
Beeswax 1-5, dodecyl sulphate 1-5, are formulated according to aforementioned proportion.
3. a kind of preparation method of arrowhead polysaccharide composition preventing chemical damage according to claim 2, special
Sign is that Subcritical Water Extraction condition is：Filtered fluid is injected into subcritical abstraction kettle through compression pump, controls magnetic-coupling stirrer
300-600 turns/min, extracting pressure 1-12MPa, and 100-180 DEG C of temperature extracts 10-30min, after, supernatant is collected in centrifugation
Liquid, as arrowhead Subcritical Water Extraction liquid.
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