CN107125515A - It is a kind of to prepare the method with anti-oxidation function bee peptide drink - Google Patents
It is a kind of to prepare the method with anti-oxidation function bee peptide drink Download PDFInfo
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- CN107125515A CN107125515A CN201710175288.XA CN201710175288A CN107125515A CN 107125515 A CN107125515 A CN 107125515A CN 201710175288 A CN201710175288 A CN 201710175288A CN 107125515 A CN107125515 A CN 107125515A
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- 230000003064 anti-oxidating effect Effects 0.000 title claims abstract description 33
- 238000000034 method Methods 0.000 title claims abstract description 24
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 16
- 241000241413 Propolis Species 0.000 claims abstract description 76
- 229940069949 propolis Drugs 0.000 claims abstract description 76
- 229930003944 flavone Natural products 0.000 claims abstract description 46
- 235000011949 flavones Nutrition 0.000 claims abstract description 46
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 claims abstract description 43
- 150000002212 flavone derivatives Chemical class 0.000 claims abstract description 43
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 claims abstract description 43
- 239000003659 bee venom Substances 0.000 claims abstract description 40
- 101000761020 Dinoponera quadriceps Poneritoxin Proteins 0.000 claims abstract description 30
- 108010036176 Melitten Proteins 0.000 claims abstract description 25
- VDXZNPDIRNWWCW-JFTDCZMZSA-N melittin Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(N)=O)CC1=CNC2=CC=CC=C12 VDXZNPDIRNWWCW-JFTDCZMZSA-N 0.000 claims abstract description 25
- 238000000605 extraction Methods 0.000 claims abstract description 14
- 238000005119 centrifugation Methods 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 36
- 239000007787 solid Substances 0.000 claims description 26
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 24
- 238000002360 preparation method Methods 0.000 claims description 19
- 235000020097 white wine Nutrition 0.000 claims description 19
- 230000003078 antioxidant effect Effects 0.000 claims description 17
- 239000003963 antioxidant agent Substances 0.000 claims description 16
- 235000006708 antioxidants Nutrition 0.000 claims description 16
- 229920005654 Sephadex Polymers 0.000 claims description 15
- 239000012507 Sephadex™ Substances 0.000 claims description 15
- 230000001476 alcoholic effect Effects 0.000 claims description 15
- 238000000926 separation method Methods 0.000 claims description 15
- 239000000284 extract Substances 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 12
- 230000005526 G1 to G0 transition Effects 0.000 claims description 10
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims description 10
- 238000010828 elution Methods 0.000 claims description 9
- 230000000844 anti-bacterial effect Effects 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 6
- 108010024636 Glutathione Proteins 0.000 claims description 5
- 108010053070 Glutathione Disulfide Proteins 0.000 claims description 5
- 229920001231 Polysaccharide peptide Polymers 0.000 claims description 5
- 238000004132 cross linking Methods 0.000 claims description 5
- 238000004821 distillation Methods 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- 238000009826 distribution Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- 235000019441 ethanol Nutrition 0.000 claims description 5
- 238000007710 freezing Methods 0.000 claims description 5
- 230000008014 freezing Effects 0.000 claims description 5
- 229960003180 glutathione Drugs 0.000 claims description 5
- YPZRWBKMTBYPTK-BJDJZHNGSA-N glutathione disulfide Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@H](C(=O)NCC(O)=O)CSSC[C@@H](C(=O)NCC(O)=O)NC(=O)CC[C@H](N)C(O)=O YPZRWBKMTBYPTK-BJDJZHNGSA-N 0.000 claims description 5
- 238000002386 leaching Methods 0.000 claims description 5
- 230000014759 maintenance of location Effects 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- 108010022457 polysaccharide peptide Proteins 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 230000002829 reductive effect Effects 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims 1
- 150000004676 glycans Chemical class 0.000 claims 1
- 230000003647 oxidation Effects 0.000 abstract description 7
- 238000007254 oxidation reaction Methods 0.000 abstract description 7
- 235000013305 food Nutrition 0.000 abstract description 4
- 230000032683 aging Effects 0.000 abstract description 2
- 238000010266 Sephadex chromatography Methods 0.000 abstract 1
- 239000011149 active material Substances 0.000 abstract 1
- 230000036039 immunity Effects 0.000 abstract 1
- 229920001184 polypeptide Polymers 0.000 abstract 1
- 102000004196 processed proteins & peptides Human genes 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 17
- 150000003254 radicals Chemical class 0.000 description 8
- -1 DPPH free radical Chemical class 0.000 description 7
- 238000013329 compounding Methods 0.000 description 6
- 230000002000 scavenging effect Effects 0.000 description 5
- 229920001503 Glucan Polymers 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 241000256844 Apis mellifera Species 0.000 description 3
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 3
- 150000002213 flavones Chemical class 0.000 description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 230000000840 anti-viral effect Effects 0.000 description 2
- 238000009341 apiculture Methods 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- 230000007760 free radical scavenging Effects 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000000469 ethanolic extract Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 235000020510 functional beverage Nutrition 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000000724 leishmaniacidal effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 150000002926 oxygen Chemical class 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
Classifications
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
-
- 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
-
- 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
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Jellies, Jams, And Syrups (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The method with anti-oxidation function bee peptide drink is prepared the invention discloses a kind of, belongs to biological technical field.The anti-oxidation function drink is the complex configuration of bee venom peptide solution and propolis flavone solution, melittin is that the polypeptide got is isolated and purified from bee venom, propolis flavone is the active material extracted from propolis, crushed using sephadex chromatography, high speed, the method taken with reference to centrifugation, temperature extraction, obtains bee venom peptide solution and propolis flavone solution.The oxidation resistance functional form food of the present invention, with good antioxidation, and is unlikely to deteriorate, thus can deposit for a long time.The anti-oxidation function type food prepared by the present invention is drunk, reducing human aging and immunity of organisms can be improved.
Description
Technical field
The method with anti-oxidation function drink is prepared the present invention relates to a kind of, belongs to biological technical field.
Background technology
With the improvement of living standards, the food demand with anti-oxidation function is growing day by day, various anti-oxidant productions are utilized
Product remove the free radical produced in vivo, can reach the aging for slowing down body, reach the purpose of health care.Propolis is worker bee
The gumminess material that the secretion such as the secretion such as herborization resin and its mandibular gland, wax gland is mixed to form, containing a variety of natural
Effective active matter, such as flavones, phenolic acid class and terpenoid substance, it is immune with anti-oxidant, antibacterial, antiviral, anticancer, enhancing
A variety of effects such as power, because it has the advantages that biocidal property, anti-corrosive properties, have been widely used in the neck such as food, medicine, cosmetics
Domain.Melittin is the main component of bee venom, accounts for the 50% of bee venom dry weight, with radioresistance, anti-inflammation, antiviral, antitumor
Deng effect, there is certain application in the field such as medicine and cosmetics.
Propolis flavone is extracted from propolis virgin rubber and got, with it is easy extract, the cycle is short the features such as, melittin is from honeybee
Isolate and purify what is got in poison, with it is easy prepare, low amounts is efficient the features such as, it is anti-oxidant that both combinations can develop body
Deng product, with the very big prospect of marketing, thus cause academia and widely pay close attention to.
The preparation of oxidation resistant product, can use extraction method and synthetic method, and artificial synthesized oxidation resistant product has certain pair
Effect, can threaten health.Mouhoubi-Tafinine and Yong（1. Mouhoubi-Tafinine, Z., et al.,
Antioxydant activity of some algerian honey and propolis[J]. Industrial Crops
& Products, 2016, 88: 85-90. 2. Yong, K.P. and M. Ikegaki, Preparation of
Water and Ethanolic Extracts of Propolis and Evaluation of the Preparations
[J]. Bioscience Biotechnology & Biochemistry, 1998, 62(11): 2230-2232.）Report
The antioxidation activity of beta carotene-linoleic acid colorimetric method for determining propolis flavone, it was demonstrated that propolis flavone has oxidation resistant effect.
Guo and Nascimento（Guo, X., et al., Chemical Compositions and Antioxidant
Activities of Water Extracts of Chinese Propolis[J]. Biochemistry, 2012, 59
(23): 12610-12616. 2. Ticiano Gomes do Nascimento, et al., Polymeric
Nanoparticles of Brazilian Red Propolis Extract: Preparation,
Characterization, Antioxidant and Leishmanicidal Activity[J].Nanoscale
Research Letters, 2016, 11: 301.）Report and remove the anti-oxidant work that DPPH free radical methods determine propolis flavone
Property, IC50 values are 38.0 μ g/mL, but antioxidation activity is relatively low.Cao little Yan（Cao little Yan etc., the extraction of Flavonoids in Propolis and its
Free radical scavenging activity research [J] oil and foodstuffs science and technology, 2015,23 (5): 45-49.）Report scavenging hydroxyl energy
Amylograph and scavenging activated oxygen energy amylograph determine the antioxidation activity of propolis flavone, 0.15mg/mL propolis flavones pair
The clearance rate of hydroxyl radical free radical and ultra-oxygen anion free radical has respectively reached 35%（Mass fraction）With 77. 7%（Quality point
Number）, but the antioxidation activity of propolis flavone is relatively low, Wu Zhenhong（Wu Zhenhong etc., scavenging action of the melittin to free radical
[J] China's apicultures, 2007,58 (5): 9-10.）Report the antioxygen that scavenging hydroxyl energy amylograph determines melittin
Change activity, 66.7 μ g/mL melittin is 67.39% to the clearance rate of hydroxyl radical free radical（Mass fraction）, but melittin is to hydroxyl
The clearance rate of base free radical is low, and antioxidation activity is relatively low.
The above method is the anti-oxidant of one-component, and antioxidation activity is low.
The content of the invention
The invention aims to solve the problem of single component drink antioxidation activity is low, there is provided a kind of anti-oxidant work
The high functional beverage of property.
The concentration that the present invention is obtained be 0.2mg/mL bee venom peptide solution with concentration be 0.83mg/mL propolis flavones solution according to
Volume ratio V（Melittin）：V（Propolis）=1:5 are compounded, and compounding drink is to hydroxyl radical free radical and ultra-oxygen anion free radical
Clearance rate has respectively reached 86.31%（Mass fraction）With 94.52%（Mass fraction）.
A kind of compounding drink with anti-oxidation function of the present invention, step is as follows：
1. the preparation of propolis flavone solution
Propolis is put into the white wine that alcoholic strength is 45-75 °, 70-90 DEG C of extraction 1.5-4.5h is extracted 1-3 times, with 2500- altogether
5500r/min centrifugations 4-8min carries out separation of solid and liquid, obtains propolis flavone extract solution and propolis residue, is 45-75 ° with alcoholic strength
White wine by propolis flavone extract solution be configured to concentration be 0.5-1.0mg/mL propolis flavone solution；
The alcoholic strength is the percent by volume of contained ethanol in white wine；
It is described to be extracted as taking in the middle temperature extraction of water-bath；
The separation of solid and liquid is the process that propolis residue is removed from leaching liquor；
The white wine is food-grade.
2. the preparation of bee venom peptide solution
By bee venom through the distillation water elution of sephadex G -25, flow control determines point of each separation component in 25-30mL/h
Son amount, collects molecular weight for 2840Da chromatographic fraction under sephadex column and is freeze-dried to obtain solid sample, by solid
Sample is dissolved with the absolute ethyl alcohol of 2-4 times of volume, centrifuges 4-6min through 2500-3500r/min, deposit is bee venom
Peptide, the bee venom peptide solution that concentration is 0.1-0.3mg/mL is configured to pure water by melittin；
The sephadex G -25 is that glucan is represented with English alphabet G, the G reflections crosslinking degree of gel, degrees of expansion and
Distribution, 25 absorb water 2.5 grams when being every gram of gel expansion；
The elution is that chromatographic column fixed phase is continued through using distilled water as mobile phase, mobile phase and stationary phase force ratio sample
Product are weak, and sample each component is sequentially washed out from stationary phase；
The Da is the unit of bee venom peptide molecular weight；
Being determined as the molecular weight elutes retention time T (min) molecular mass logarithm values corresponding thereto according to standard items
(lgMr) standard curve regression equation, standard items are antibacterial poly saccharide peptide standard product（Mr =1422Da）, GSSG standard items
(Mr=612Da), reductive glutathione standard items (Mr=307.32Da)；
Obtained component is is refrigerated to the below freezing of water by the freeze-drying, in the container for being placed in high vacuum (10-40Pa),
The moisture in material is set directly to be distilled from solid ice as a kind of drying means of steam by heat supply；
The solid matter being precipitated as insoluble in absolute ethyl alcohol；
The pure water is food-grade.
3. the preparation of anti-oxidant drink
1. middle concentration the propolis flavone solution for being 0.5-1.0mg/mL and melittin that 2. middle concentration is 0.1-0.3mg/mL is molten
Liquid is according to volume ratio V（Melittin）：V（Propolis）=1:3-6 is compounded, and gained is the drink with anti-oxidation function.
The volume ratio is the ratio of bee venom peptide solution and propolis flavone liquor capacity；
The compounding is the mixing of bee venom peptide solution and propolis flavone solution.
Oxidation resistant drink is produced using this method, compared with the conventional method antioxidation activity increase 15-20%,
Compared with prior art, the method that the present invention prepares antioxygen drink with function, with following distinguishing feature：
（1）Propolis has anti-oxidant, reduction blood fat, immunological regulation, the effect of anti-inflammatory sterilization；
（2）Melittin, which has, removes free radical, radioresistance, the effect of anti-inflammatory sterilization, antineoplastic action；
（3）The drink oxidation resistance that the present invention is obtained is higher than vitamin C by 20% or so, be it is a kind of it is good have it is oxidation resistant
Functional food.
（4）The drink that the present invention is obtained is relative to Cao little Yan, Wu Zhenhong（1. Cao little Yan etc., the extraction of Flavonoids in Propolis and
Its free radical scavenging activity research [J] oil and foodstuffs science and technology, 2015,23 (5):The Wu Zhen of 45-49. 2. are red etc., melittin
To scavenging action [J] China's apicultures of free radical, 2007,58 (5): 9-10.）Antioxidation activity improves 15-20%.
Embodiment 1：
1. the preparation of propolis flavone solution
Propolis is put into the white wine that alcoholic strength is 60 °, 80 DEG C of extraction 3h are extracted 1 time altogether, are entered with 5000r/min centrifugations 5min
Row separation of solid and liquid, obtains propolis flavone extract solution and propolis residue, is matched somebody with somebody propolis flavone extract solution for 60 ° of white wine with alcoholic strength
It is set to the propolis flavone solution that concentration is 0.83mg/mL；
The alcoholic strength is the percent by volume of contained ethanol in white wine；
It is described to be extracted as taking in the middle temperature extraction of water-bath；
The separation of solid and liquid is the process that propolis residue is removed from leaching liquor；
The white wine is food-grade.
2. the preparation of bee venom peptide solution
By bee venom through the distillation water elution of sephadex G -25, flow control determines the molecule of each separation component in 28mL/h
Amount, collects molecular weight for 2840Da chromatographic fraction under sephadex column and is freeze-dried to obtain solid sample, by solid-like
Product are dissolved with the absolute ethyl alcohol of 3 times of volumes, centrifuge 5min through 3000r/min, deposit is melittin, by melittin
The bee venom peptide solution that concentration is 0.2mg/mL is configured to pure water；
The sephadex G -25 is that glucan is represented with English alphabet G, the G reflections crosslinking degree of gel, degrees of expansion and
Distribution, 25 absorb water 2.5 grams when being every gram of gel expansion；
The elution is that chromatographic column fixed phase is continued through using distilled water as mobile phase, mobile phase and stationary phase force ratio sample
Product are weak, and sample each component is sequentially washed out from stationary phase；
The Da is the unit of bee venom peptide molecular weight；
Being determined as the molecular weight elutes retention time T (min) molecular mass logarithm values corresponding thereto according to standard items
(lgMr) standard curve regression equation, standard items are antibacterial poly saccharide peptide standard product（Mr =1422Da）, GSSG standard items
(Mr=612Da), reductive glutathione standard items (Mr=307.32Da)；
Obtained component is is refrigerated to the below freezing of water by the freeze-drying, in the container for being placed in high vacuum (10-40Pa),
The moisture in material is set directly to be distilled from solid ice as a kind of drying means of steam by heat supply；
The solid matter being precipitated as insoluble in absolute ethyl alcohol；
The pure water is food-grade.
3. the preparation of anti-oxidant drink
By 1. middle concentration the propolis flavone solution for being 0.83mg/mL and bee venom peptide solution that 2. middle concentration is 0.2mg/mL according to body
Product compares V（Melittin）：V（Propolis）=1:5 are compounded, and gained is the drink with anti-oxidation function.
The volume ratio is the ratio of bee venom peptide solution and propolis flavone liquor capacity；
The compounding is the mixing of bee venom peptide solution and propolis flavone solution.
It is 86.31% to hydroxyl radical free radical clearance rate mass fraction that present invention, which obtains anti-oxidant drink, to superoxide anion
The clearance rate mass fraction of free radical is 94.52%.
Embodiment 2：
1. the preparation of propolis flavone solution
Propolis is put into the white wine that alcoholic strength is 70 °, 90 DEG C of extraction 4h are extracted 2 times altogether, are entered with 4000r/min centrifugations 6min
Row separation of solid and liquid, obtains propolis flavone extract solution and propolis residue, is matched somebody with somebody propolis flavone extract solution for 70 ° of white wine with alcoholic strength
It is set to the propolis flavone solution that concentration is 0.996mg/mL；
The alcoholic strength is the percent by volume of contained ethanol in white wine；
It is described to be extracted as taking in the middle temperature extraction of water-bath；
The separation of solid and liquid is the process that propolis residue is removed from leaching liquor；
The white wine is food-grade.
2. the preparation of bee venom peptide solution
By bee venom through the distillation water elution of sephadex G -25, flow control determines the molecule of each separation component in 30mL/h
Amount, collects molecular weight for 2840Da chromatographic fraction under sephadex column and is freeze-dried to obtain solid sample, by solid-like
Product are dissolved with the absolute ethyl alcohol of 4 times of volumes, centrifuge 6min through 3200r/min, deposit is melittin, by melittin
The bee venom peptide solution that concentration is 0.25mg/mL is configured to pure water；
The sephadex G -25 is that glucan is represented with English alphabet G, the G reflections crosslinking degree of gel, degrees of expansion and
Distribution, 25 absorb water 2.5 grams when being every gram of gel expansion；
The elution is that chromatographic column fixed phase is continued through using distilled water as mobile phase, mobile phase and stationary phase force ratio sample
Product are weak, and sample each component is sequentially washed out from stationary phase；
The Da is the unit of bee venom peptide molecular weight；
Being determined as the molecular weight elutes retention time T (min) molecular mass logarithm values corresponding thereto according to standard items
(lgMr) standard curve regression equation, standard items are antibacterial poly saccharide peptide standard product（Mr =1422Da）, GSSG standard items
(Mr=612Da), reductive glutathione standard items (Mr=307.32Da)；
Obtained component is is refrigerated to the below freezing of water by the freeze-drying, in the container for being placed in high vacuum (10-40Pa),
The moisture in material is set directly to be distilled from solid ice as a kind of drying means of steam by heat supply；
The solid matter being precipitated as insoluble in absolute ethyl alcohol；
The pure water is food-grade.
3. the preparation of anti-oxidant drink
By 1. middle concentration the propolis flavone solution for being 0.996mg/mL and bee venom peptide solution that 2. middle concentration is 0.25mg/mL according to
Volume ratio V（Melittin）：V（Propolis）=1:6 are compounded, and gained is the drink with anti-oxidation function.
The volume ratio is the ratio of bee venom peptide solution and propolis flavone liquor capacity；
The compounding is the mixing of bee venom peptide solution and propolis flavone solution.
It is 86.21% to hydroxyl radical free radical clearance rate mass fraction that present invention, which obtains anti-oxidant drink, to superoxide anion
The clearance rate mass fraction of free radical is 94.30%.
Embodiment 3：
1. the preparation of propolis flavone solution
Propolis is put into the white wine that alcoholic strength is 50 °, 70 DEG C of extraction 2h are extracted 3 times altogether, are entered with 3000r/min centrifugations 4min
Row separation of solid and liquid, obtains propolis flavone extract solution and propolis residue, is matched somebody with somebody propolis flavone extract solution for 50 ° of white wine with alcoholic strength
It is set to the propolis flavone solution that concentration is 0.664mg/mL；
The alcoholic strength is the percent by volume of contained ethanol in white wine；
It is described to be extracted as taking in the middle temperature extraction of water-bath；
The separation of solid and liquid is the process that propolis residue is removed from leaching liquor；
The white wine is food-grade.
2. the preparation of bee venom peptide solution
By bee venom through the distillation water elution of sephadex G -25, flow control determines the molecule of each separation component in 26mL/h
Amount, collects molecular weight for 2840Da chromatographic fraction under sephadex column and is freeze-dried to obtain solid sample, by solid-like
Product are dissolved with the absolute ethyl alcohol of 2 times of volumes, centrifuge 4min through 2800r/min, deposit is melittin, by melittin
The bee venom peptide solution that concentration is 0.15mg/mL is configured to pure water；
The sephadex G -25 is that glucan is represented with English alphabet G, the G reflections crosslinking degree of gel, degrees of expansion and
Distribution, 25 absorb water 2.5 grams when being every gram of gel expansion；
The elution is that chromatographic column fixed phase is continued through using distilled water as mobile phase, mobile phase and stationary phase force ratio sample
Product are weak, and sample each component is sequentially washed out from stationary phase；
The Da is the unit of bee venom peptide molecular weight；
Being determined as the molecular weight elutes retention time T (min) molecular mass logarithm values corresponding thereto according to standard items
(lgMr) standard curve regression equation, standard items are antibacterial poly saccharide peptide standard product（Mr =1422Da）, GSSG standard items
(Mr=612Da), reductive glutathione standard items (Mr=307.32Da)；
Obtained component is is refrigerated to the below freezing of water by the freeze-drying, in the container for being placed in high vacuum (10-40Pa),
The moisture in material is set directly to be distilled from solid ice as a kind of drying means of steam by heat supply；
The solid matter being precipitated as insoluble in absolute ethyl alcohol；
The pure water is food-grade.
3. the preparation of anti-oxidant drink
By 1. middle concentration the propolis flavone solution for being 0.664mg/mL and bee venom peptide solution that 2. middle concentration is 0.15mg/mL according to
Volume ratio V（Melittin）：V（Propolis）=1:4 are compounded, and gained is the drink with anti-oxidation function.
The volume ratio is the ratio of bee venom peptide solution and propolis flavone liquor capacity；
The compounding is the mixing of bee venom peptide solution and propolis flavone solution.
It is 82.84% to hydroxyl radical free radical clearance rate mass fraction that present invention, which obtains anti-oxidant drink, to superoxide anion
The clearance rate mass fraction of free radical is 90.06%.
Claims (10)
1. a kind of prepare the method with anti-oxidation function bee peptide drink, step is as follows：
1. the preparation of propolis flavone solution
Propolis is put into the white wine that alcoholic strength is 45-75 °, 70-90 DEG C of extraction 1.5-4.5h is extracted 1-3 times, with 2500- altogether
5500r/min centrifugations 4-8min carries out separation of solid and liquid, obtains propolis flavone extract solution and propolis residue, is 45-75 ° with alcoholic strength
White wine by propolis flavone extract solution be configured to concentration be 0.5-1.0mg/mL propolis flavone solution；
2. the preparation of bee venom peptide solution
By bee venom through the distillation water elution of sephadex G -25, flow control determines point of each separation component in 25-30mL/h
Son amount, collects molecular weight for 2840Da chromatographic fraction under sephadex column and is freeze-dried to obtain solid sample, by solid
Sample is dissolved with the absolute ethyl alcohol of 2-4 times of volume, centrifuges 4-6min through 2500-3500r/min, deposit is bee venom
Peptide, the bee venom peptide solution that concentration is 0.1-0.3mg/mL is configured to pure water by melittin；
3. the preparation of anti-oxidant drink
1. middle concentration the propolis flavone solution for being 0.5-1.0mg/mL and melittin that 2. middle concentration is 0.1-0.3mg/mL is molten
Liquid is according to volume ratio V（Melittin）：V（Propolis）=1:3-6 is compounded, and gained is the drink with anti-oxidation function.
2. as claimed in claim 1 prepare the method with anti-oxidation function bee peptide drink, alcoholic strength is contained in white wine
The percent by volume of food-grade ethanol.
3. as claimed in claim 1 prepare the method with anti-oxidation function bee peptide drink, it is extracted as in the middle temperature of water-bath
Extraction takes.
4. as claimed in claim 1 prepare the method with anti-oxidation function bee peptide drink, separation of solid and liquid is from leaching liquor
The middle process for removing propolis residue.
5. as claimed in claim 1 prepare the method with anti-oxidation function bee peptide drink, sephadex G -25 is Portugal
Glycan represents with English alphabet G, and G reflects crosslinking degree, degrees of expansion and the distribution of gel, and 25 when being every gram of gel expansion
2.5 grams of water suction.
6. as claimed in claim 1 prepare the method with anti-oxidation function bee peptide drink, elute for using distilled water as
Mobile phase continues through chromatographic column fixed phase, and mobile phase and stationary phase force ratio sample are weak, and sample each component is sequentially
Washed out from stationary phase.
7. as claimed in claim 1 prepare the method with anti-oxidation function bee peptide drink, Da is bee venom peptide molecular weight
Unit.
8. as claimed in claim 1 prepare the method with anti-oxidation function bee peptide drink, molecular weight is determined as basis
Standard items elute retention time T (min) molecular mass logarithm value (lgMr) standard curve regression equations corresponding thereto, and standard items are
Antibacterial poly saccharide peptide standard product（Mr =1422Da）, GSSG standard items (Mr=612Da), reductive glutathione standard items
(Mr =307.32Da)。
9. as claimed in claim 1 prepare the method with anti-oxidation function bee peptide drink, it is freeze-dried as by gained group
Divide and be refrigerated to the below freezing of water, in the container for being placed in high vacuum (10-40Pa), make the moisture in material direct by heat supply
Distilled from solid ice as a kind of drying means of steam.
10. as claimed in claim 1 prepare the method with anti-oxidation function bee peptide drink, compound for bee venom peptide solution with
The mixing of propolis flavone solution.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710175288.XA CN107125515A (en) | 2017-03-22 | 2017-03-22 | It is a kind of to prepare the method with anti-oxidation function bee peptide drink |
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| CN201710175288.XA CN107125515A (en) | 2017-03-22 | 2017-03-22 | It is a kind of to prepare the method with anti-oxidation function bee peptide drink |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1544429A (en) * | 2003-11-21 | 2004-11-10 | 长春中医学院 | Bee glue flavone extract preparation method, pharmaceutical preparation and its new medical uses |
| CN101089017A (en) * | 2006-06-16 | 2007-12-19 | 张文礼 | Process of separating and purifying melittin |
| CN102526116A (en) * | 2011-11-23 | 2012-07-04 | 程文显 | Method for refining bee venom |
-
2017
- 2017-03-22 CN CN201710175288.XA patent/CN107125515A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1544429A (en) * | 2003-11-21 | 2004-11-10 | 长春中医学院 | Bee glue flavone extract preparation method, pharmaceutical preparation and its new medical uses |
| CN101089017A (en) * | 2006-06-16 | 2007-12-19 | 张文礼 | Process of separating and purifying melittin |
| CN102526116A (en) * | 2011-11-23 | 2012-07-04 | 程文显 | Method for refining bee venom |
Non-Patent Citations (2)
| Title |
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| 刘䶮 等: ""蜂胶黄酮提取纯化的工艺研究"", 《中国蜂业》 * |
| 杨文超: "蜂毒肽的分离纯化及其抗辐射作用机理研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
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