CN106188276A - A kind of preparation method of albumen catechin free radical grafting thing - Google Patents
A kind of preparation method of albumen catechin free radical grafting thing Download PDFInfo
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- CN106188276A CN106188276A CN201610540133.7A CN201610540133A CN106188276A CN 106188276 A CN106188276 A CN 106188276A CN 201610540133 A CN201610540133 A CN 201610540133A CN 106188276 A CN106188276 A CN 106188276A
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- Prior art keywords
- catechin
- albumen
- free radical
- sample
- radical grafting
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- 229950001002 cianidanol Drugs 0.000 title claims abstract description 53
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 title claims abstract description 26
- 235000005487 catechin Nutrition 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- -1 catechin free radical Chemical class 0.000 title abstract description 7
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 36
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 34
- 108010000912 Egg Proteins Proteins 0.000 claims abstract description 33
- 102000002322 Egg Proteins Human genes 0.000 claims abstract description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 30
- 239000000243 solution Substances 0.000 claims abstract description 29
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-catechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 claims abstract description 24
- 238000003756 stirring Methods 0.000 claims abstract description 20
- 241000287828 Gallus gallus Species 0.000 claims abstract description 19
- 210000004681 ovum Anatomy 0.000 claims abstract description 19
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 19
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 19
- 235000010323 ascorbic acid Nutrition 0.000 claims abstract description 15
- 239000011668 ascorbic acid Substances 0.000 claims abstract description 15
- 229960005070 ascorbic acid Drugs 0.000 claims abstract description 15
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims abstract description 14
- 235000014103 egg white Nutrition 0.000 claims abstract description 14
- 210000000969 egg white Anatomy 0.000 claims abstract description 14
- 238000004108 freeze drying Methods 0.000 claims abstract description 14
- 239000004615 ingredient Substances 0.000 claims abstract description 12
- 238000004062 sedimentation Methods 0.000 claims abstract description 12
- 238000010790 dilution Methods 0.000 claims abstract description 10
- 239000012895 dilution Substances 0.000 claims abstract description 10
- 235000013305 food Nutrition 0.000 claims abstract description 8
- 239000000523 sample Substances 0.000 claims description 33
- 238000006243 chemical reaction Methods 0.000 claims description 13
- 150000003254 radicals Chemical class 0.000 claims description 11
- 239000003999 initiator Substances 0.000 claims description 9
- 238000000502 dialysis Methods 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 7
- 239000012488 sample solution Substances 0.000 claims description 3
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims description 2
- 229930003268 Vitamin C Natural products 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 235000019154 vitamin C Nutrition 0.000 claims description 2
- 239000011718 vitamin C Substances 0.000 claims description 2
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 abstract description 12
- 150000008442 polyphenolic compounds Chemical class 0.000 abstract description 12
- 235000013824 polyphenols Nutrition 0.000 abstract description 12
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 abstract description 5
- 230000007760 free radical scavenging Effects 0.000 abstract description 5
- 238000005516 engineering process Methods 0.000 abstract description 3
- 235000018102 proteins Nutrition 0.000 description 17
- 229920000159 gelatin Polymers 0.000 description 14
- 235000019322 gelatine Nutrition 0.000 description 14
- 108010010803 Gelatin Proteins 0.000 description 13
- 239000008273 gelatin Substances 0.000 description 13
- 235000011852 gelatine desserts Nutrition 0.000 description 13
- 230000008859 change Effects 0.000 description 12
- 230000031700 light absorption Effects 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- 239000002994 raw material Substances 0.000 description 7
- 230000003993 interaction Effects 0.000 description 5
- 230000003064 anti-oxidating effect Effects 0.000 description 4
- 239000012496 blank sample Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- OCZVHBZNPVABKX-UHFFFAOYSA-N 1,1-diphenyl-2-(2,4,6-trinitrophenyl)hydrazine;ethanol Chemical compound CCO.[O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1NN(C=1C=CC=CC=1)C1=CC=CC=C1 OCZVHBZNPVABKX-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108010058846 Ovalbumin Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 235000013601 eggs Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 229940092253 ovalbumin Drugs 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- GDSOZVZXVXTJMI-SNAWJCMRSA-N (e)-1-methylbut-1-ene-1,2,4-tricarboxylic acid Chemical compound OC(=O)C(/C)=C(C(O)=O)\CCC(O)=O GDSOZVZXVXTJMI-SNAWJCMRSA-N 0.000 description 1
- 244000080767 Areca catechu Species 0.000 description 1
- 235000006226 Areca catechu Nutrition 0.000 description 1
- 108010026206 Conalbumin Proteins 0.000 description 1
- 239000001828 Gelatine Substances 0.000 description 1
- 108010063045 Lactoferrin Proteins 0.000 description 1
- 102000010445 Lactoferrin Human genes 0.000 description 1
- 102000008192 Lactoglobulins Human genes 0.000 description 1
- 108010060630 Lactoglobulins Proteins 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 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
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000007334 copolymerization reaction Methods 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000010559 graft polymerization reaction Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- CSSYQJWUGATIHM-IKGCZBKSSA-N l-phenylalanyl-l-lysyl-l-cysteinyl-l-arginyl-l-arginyl-l-tryptophyl-l-glutaminyl-l-tryptophyl-l-arginyl-l-methionyl-l-lysyl-l-lysyl-l-leucylglycyl-l-alanyl-l-prolyl-l-seryl-l-isoleucyl-l-threonyl-l-cysteinyl-l-valyl-l-arginyl-l-arginyl-l-alanyl-l-phenylal Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CSSYQJWUGATIHM-IKGCZBKSSA-N 0.000 description 1
- 229940078795 lactoferrin Drugs 0.000 description 1
- 235000021242 lactoferrin Nutrition 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000012966 redox initiator Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/77—Ovalbumin
-
- 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
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses the preparation method of a kind of albumen catechin free radical grafting thing, belong to food technology field.The inventive method is the water dilution by adding 9 times of volumes in Ovum Gallus domesticus album, and final albumen quality mark is 1%, regulates pH to about 5.0, stirs 10 20min, natural sedimentation 1h, then filters and remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask, add 0.5 2.0mL 5M hydrogen peroxide and 0.0625 0.5g ascorbic acid, stir rearmounted 2h at room temperature, add 0.025 0.1g catechin, 12 48h are reacted under the conditions of 25 DEG C, dialyse under the conditions of 4 DEG C 12 60h again to sample, changes water once to guarantee that unreacted free polyphenol is dialysed away completely every 6h, obtains sample after last lyophilization.The albumen catechin free radical grafting thing obtained by the inventive method, its DPPH free radical scavenging activity is up to 65.29%, and ABTS free radical scavenging activity is 94.17%, and reducing power reaches as high as 0.649Abs.
Description
Technical field
The present invention relates to the preparation method of a kind of albumen-catechin free radical grafting thing, belong to food technology field.
Background technology
Along with the development of food industry, the demand of the protein with specific functional features and nutritive peculiarity is also existed
Constantly improve.Albumen has nutrition and the organoleptic attribute of excellence, is widely used in food.By physics, chemistry with
And biological means gives albumen and has non-oxidizability and more preferable functional character, can be widened it further at food, biology
With the application in medicine and other fields.Catechin is the general name that in tea polyphenols, a class has highly active flavonoid, has the strongest
Anti-oxidation function.But prepare albumen-catechin complex, protein and polyphenol phase interaction by mixing method or alkaline process
With more weak, it is not enough to significantly improve the non-oxidizability of protein.Therefore need badly and use new technological means promotion protein with many
Interaction between phenol molecule, forms the complex with strong anti-oxidation.
Graft polymerization technique is usually used in developing the new material with specific physicochemical structure, can give naturally occurring or synthetic high score
The functional character that sub-material is new, thus widen its application in each field further.The purpose of glycerol polymerization is some to be had
The small-molecule substance of specific functional features is grafted on macromolecular structure obtain novel complexes, has the functional of two kinds of materials concurrently
Matter.In Research on Graft copolymerization, the selection of initiator is the key of polymerization, in numerous initiators, a kind of without metallic element
Redox initiation system ascorbic acid one by one and hydrogen peroxide system the most progressively cause the attention of people.The operation letter of this system
Single, mild condition, can carry out under room temperature, and harmful by-products will not be produced, at the food neck harsh to security requirement
Territory tool is of great significance.
Ascorbic acid and hydrogen peroxide system are in terms of synthetic proteins-polyphenol complex, at present also in basic research
Starting stage, under conditions of free radical exists, interaction mechanism and this complex between protein and polyphenol should in reality
With researchs such as the effects in system seldom, the most also there is no Patents.Existing research mainly use lactoferrin,
The single albumen such as beta lactoglobulin, ovotransferrin is primary raw material.But this kind of protein raw materials price is high, source rareness, produce
Cost is the highest, limits its application in actual production.Additionally, due to albumen source is different, the interaction with polyphenol is also
The most different, the performance how improving albumen-polyphenol complex product is also current urgent problem.
Summary of the invention
In order to solve the problems referred to above, the method that the invention provides the non-oxidizability of simple and quick raising albumen, be
A kind of preparation method of albumen-catechin free radical grafting thing.The inventive method, with albumen as raw material, adds anti-bad
Hematic acid and hydrogen peroxide system are as radical initiator, and protein molecular forms bioactive molecule under hydroxyl radical free radical effect, has
It is beneficial to form the albumen-catechin free radical grafting thing with strong anti-oxidation with catechin small molecular phase interaction.And
And albumen has the advantages such as raw material sources are extensive, cheap, nutritious, market prospect is extensive.
The preparation method of the albumen of the present invention-catechin free radical grafting thing, described method includes: the most first pretreatment
Ovum Gallus domesticus album, to remove the mucin in Ovum Gallus domesticus album, adds radical initiator in egg white solution, then adds catechin and forms Ovum Gallus domesticus album
Albumen-catechin free radical grafting thing, then it is the completeest to guarantee unreacted free catechin that sample solution carries out dialysis treatment
Full removal, obtains product after last lyophilization.
In one embodiment of the invention, described Ovum Gallus domesticus album pretreatment is: the water adding 9 times of volumes in Ovum Gallus domesticus album is dilute
Releasing, final albumen quality mark is 1%, regulates pH to about 5.0, stirs 10-20min, natural sedimentation 1h, then filters removal
Insoluble protein ingredient.
In one embodiment of the invention, the mass ratio of the albumen in described catechin and egg white solution be 1:20～
1:5。
In one embodiment of the invention, final concentration of the 0.5～2g/L of described catechin.
In one embodiment of the invention, described radical initiator is hydrogen peroxide and ascorbic acid.
In one embodiment of the invention, final concentration of the 0.05 of described hydrogen peroxide～0.2mol/L, Vitamin C
Final concentration of the 1.25～10g/L of acid.
In one embodiment of the invention, the volume of described albumen solution is 50mL；Described free radical causes
Agent is 0.5-2.0mL 5M hydrogen peroxide and 0.0625-0.5g ascorbic acid.
In one embodiment of the invention, described radical initiator is 25 DEG C with the reaction condition of albumen,
2h。
In one embodiment of the invention, the addition of described catechin is 0.025-0.1g.
In one embodiment of the invention, the reaction condition of described catechin and albumen is 25 DEG C, 12～
48h。
In one embodiment of the invention, described dialysis treatment is to continue under the conditions of 4 DEG C.
In one embodiment of the invention, described dialysis treatment is dialysis 12-60h, changes water once every 6h.
In one embodiment of the invention, described preparation method, specifically include: in Ovum Gallus domesticus album, add 9 times of volumes
Water dilutes, and final albumen quality mark is 1%, regulates pH to about 5.0, stirs 10-20min, natural sedimentation 1h, then filters
Remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask, add 0.5-2.0mL 5M hydrogen peroxide and
0.0625-0.5g ascorbic acid, stir rearmounted 2h at room temperature, adds 0.025-0.1g catechin, under the conditions of 25 DEG C
Reaction 12-48h, then sample carries out at 4 DEG C dialysis 12-60h, and (if room temperature condition is dialysed, more than 24h, albumen will
Degeneration precipitates), change water once to guarantee that unreacted free polyphenol is dialysed away completely every 6h, obtain after last lyophilization
Sample.
The present invention is also claimed albumen-catechin free radical grafting thing that described method obtains, and described egg
Albumin-catechin free radical grafting thing application in terms of food.
Albumen prepared by the present invention-catechin free radical grafting thing need to be dialysed under the conditions of 4 DEG C.
Beneficial effects of the present invention:
(1) the free radical grafting method that the present invention uses is simple to operate, mild condition, and will not produce harmful by-products.
(2) compared with complex prepared by albumen and alkaline process, albumen prepared by the present invention-catechin free radical
Graft, its non-oxidizability is significantly improved, and its DPPH free radical scavenging activity is 65.29%, and ABTS free radical scavenging activity is
94.17%, reducing power reaches as high as 0.649Abs.
(3) preparation method of the gelatin-catechin free radical grafting thing and reported, the free radical that the present invention uses causes
The addition of agent and catechin is less, i.e. can reach the non-oxidizability of gelatin-catechin free radical grafting thing.At same process bar
Under part, the present invention uses albumen to be that albumen-catechin prepared by primary raw material has higher non-oxidizability.
Detailed description of the invention
(1) mensuration of DPPH radical scavenging activity
Weighing DPPH powder 4mg, with anhydrous alcohol solution and be settled to 100mL, being configured to ultimate density is 0.1mM's
DPPH ethanol solution, is positioned in brown bottle, and it is standby to be saved in dark place.By diluted sample suitable multiple, sample is made to carry out point
During analysis, its light absorption value is preferably as the criterion below 0.7.Take 2mL DPPH ethanol solution and 2mL sample (1mg/mL) in tool plug test tube
Mixing, opens in dark place after mixing in time, measures absorbance after 30min at 517nm, simultaneously right using dehydrated alcohol as blank
According to, experiment in triplicate, and is calculated as follows the clearance rate of DPPH free radical:
In formula, A0Blank is blank sample light absorption value at 517nm, and A is testing sample light absorption value at 517nm.
(2)ABTS+·The mensuration of Scavenging activity
7mM ABTS solution and 2.45mM potassium persulfate solution equal-volume are mixed and made into ABTS working stocks, room temperature lucifuge
Deposit 16h, be diluted with 5mM pH 7.4 phosphate buffer before using, it is desirable to the absorbance of ABTS working solution deducts phosphoric acid
After salt buffer blank, it is 0.70 ± 0.02.Accurately measure the ABTS of 2.5mL sample solution (0.5mg/mL) and 2.5mL
Working solution mixes, and measures its light absorption value under 734nm wavelength after room temperature reaction 15min, simultaneously using phosphate buffer as
Blank, parallel test three times.ABTS+·The computing formula of clearance rate is as follows:
In formula: A0For blank sample and ABTS+·Reacted light absorption value；A is sample and ABTS+·Reacted light absorption value.
(3) mensuration of iron ion reducing power
Measure 1.0mL sample liquid (10mg/mL), add phosphate buffer (pH 6.6) 2.5mL and 1% of 0.2moL/L
Potassium ferricyanide solution 2.5mL, mixture, in 50 DEG C of water bath heat preservation 30min, is subsequently adding 2.5mL 10% (w/v) trichloroacetic acid
Solution, under 3000g rotating speed, centrifugal 10min, accurately pipettes supernatant 2.5mL, is sequentially added into 2.5mL deionized water and 0.5mL
The liquor ferri trichloridi of 0.1%, surveys light absorption value (light absorption value is the biggest, and reducing power is the strongest) at 700nm after reaction 10min.To go
Ionized water is positive control.Experiment in triplicate, is averaged.
Embodiment 1: the preparation of albumen-catechin free radical grafting thing
Adding the water dilution of 9 times of volumes in Ovum Gallus domesticus album, final albumen quality mark is 1%, regulates pH to about 5.0, stirs
Mix 10-20min, natural sedimentation 1h, then filter and remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask
In, add 1.0mL 5M hydrogen peroxide and 0.25g ascorbic acid, stir rearmounted 2h at room temperature, adds 0.025g
Theine, reacts 24h, then sample carries out the 48h that dialyses under the conditions of 25 DEG C, change water every 6h once unreacted free many to guarantee
Phenol is dialysed away completely, obtains sample after last lyophilization.
Embodiment 2: the preparation of albumen-catechin free radical grafting thing
Adding the water dilution of 9 times of volumes in Ovum Gallus domesticus album, final albumen quality mark is 1%, regulates pH to about 5.0, stirs
Mix 10-20min, natural sedimentation 1h, then filter and remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask
In, add 0.5mL 5M hydrogen peroxide and 0.0625g ascorbic acid, stir rearmounted 2h at room temperature, adds 0.05g
Theine, reacts 24h, then sample carries out the 48h that dialyses under the conditions of 25 DEG C, change water every 6h once unreacted free many to guarantee
Phenol is dialysed away completely, obtains sample after last lyophilization.
Embodiment 3: the preparation of albumen-catechin free radical grafting thing
Adding the water dilution of 9 times of volumes in Ovum Gallus domesticus album, final albumen quality mark is 1%, regulates pH to about 5.0, stirs
Mix 10-20min, natural sedimentation 1h, then filter and remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask
In, add 2.0mL 5M hydrogen peroxide and 0.5g ascorbic acid, stir rearmounted 2h at room temperature, adds 0.1g catechu
Element, reacts 24h, then sample carries out the 48h that dialyses, change water once to guarantee unreacted free polyphenol every 6h under the conditions of 25 DEG C
Dialyse away completely, after last lyophilization, obtain sample.
Additionally, inventor finds, when continuing to increase the ratio of catechin and albumen, appearance in system can be caused a large amount of heavy
Forming sediment, reaction cannot proceed.Inventor has also attempted use ovalbumin
When 1%, in 100ml solution, hydrogen peroxide addition more than 1mmol or ascorbic acid addition more than 0.04g time, the white of an egg
Degeneration is precipitated by albumen, the carrying out of impact reaction.Therefore, ovalbumin is not suitable as albumen-polyphenol free radical grafting thing
Primary raw material.
Reference examples 1: the preparation of albumen blank sample
Adding the water dilution of 9 times of volumes in Ovum Gallus domesticus album, final albumen quality mark is 1%, regulates pH to about 5.0, stirs
Mix 10-20min, natural sedimentation 1h, then filter and remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask
In, react 24h under the conditions of 25 DEG C, then sample is carried out the 48h that dialyses, change water once every 6h, after last lyophilization, obtain sample
Product.
Reference examples 2: the preparation of albumen control sample
Adding the water dilution of 9 times of volumes in Ovum Gallus domesticus album, final albumen quality mark is 1%, regulates pH to about 5.0, stirs
Mix 10-20min, natural sedimentation 1h, then filter and remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask
In, add 1.0mL 5M hydrogen peroxide and 0.25g ascorbic acid, stir rearmounted 2h at room temperature, then is placed in 25 DEG C of conditions
Lower reaction 24h, then sample is carried out the 48h that dialyses, change water once every 6h, after last lyophilization, obtain sample.
Reference examples 3: the preparation of albumen-catechin alkaline process complex
Adding the water dilution of 9 times of volumes in Ovum Gallus domesticus album, final albumen quality mark is 1%, regulates pH to about 5.0, stirs
Mix 10-20min, natural sedimentation 1h, then filter and remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask
In, regulate pH to 10.0, add 0.025g catechin, react 24h under the conditions of 25 DEG C, then sample is carried out the 48h that dialyses, every 6h
Change water once to guarantee that unreacted free polyphenol is dialysed away completely, after last lyophilization, obtain sample.
Reference examples 4: the preparation of albumen-catechin alkaline process complex
Adding the water dilution of 9 times of volumes in Ovum Gallus domesticus album, final albumen quality mark is 1%, regulates pH to about 5.0, stirs
Mix 10-20min, natural sedimentation 1h, then filter and remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask
In, regulate pH to 10.0, add 0.05g catechin, react 24h under the conditions of 25 DEG C, then sample is carried out the 48h that dialyses, every 6h
Change water once to guarantee that unreacted free polyphenol is dialysed away completely, after last lyophilization, obtain sample.
Reference examples 5: the preparation of albumen-catechin alkaline process complex
Adding the water dilution of 9 times of volumes in Ovum Gallus domesticus album, final albumen quality mark is 1%, regulates pH to about 5.0, stirs
Mix 10-20min, natural sedimentation 1h, then filter and remove insoluble protein ingredient.Take 50mL egg white solution in 100mL triangular flask
In, regulate pH to 10.0, add 0.1g catechin, react 24h under the conditions of 25 DEG C, then sample is carried out the 48h that dialyses, change every 6h
Water once to guarantee that unreacted free polyphenol is dialysed away completely, obtains sample after last lyophilization.
Reference examples 6: the preparation of gelatin blank sample
Prepare 1% gelatin solution in 45 DEG C of hot water, be stirred well to gelatin and be completely dissolved.Take 50mL gelatin solution in
In 100mL triangular flask, react 24h under the conditions of 25 DEG C, then sample is carried out the 48h that dialyses, change water once every 6h, last freezing dry
Sample is obtained after dry.
Reference examples 7: the preparation of gelatin-catechin free radical grafting thing
Prepare 1% gelatin solution in 45 DEG C of hot water, be stirred well to gelatin and be completely dissolved.Take 50mL gelatin solution in
In 100mL triangular flask, adding 0.5mL 5M hydrogen peroxide and 0.0625g ascorbic acid, stir rearmounted 2h at room temperature, then
Add 0.05g catechin, react 24h under the conditions of 25 DEG C, then sample is carried out the 48h that dialyses, change water once to guarantee not every 6h
The free polyphenol of reaction is dialysed away completely, obtains sample after last lyophilization.
Reference examples 8: the preparation of gelatin-catechin free radical grafting thing
Prepare 1% gelatin solution in 45 DEG C of hot water, be stirred well to gelatin and be completely dissolved.Take 50mL gelatin solution in
In 100mL triangular flask, adding 2.0mL 5M hydrogen peroxide and 0.5g ascorbic acid, stir rearmounted 2h at room temperature, then adds
Enter 0.1g catechin, react 24h under the conditions of 25 DEG C, then sample is carried out the 48h that dialyses, change water once to guarantee unreacted every 6h
Free polyphenol dialyse away completely, obtain sample after last lyophilization.
Additionally, inventor also finds, when gelatin is as primary raw material, when gelatine content is higher than 1%, system temperature is room
Wen Shi, in course of reaction, gelatin will form gel, the carrying out of impact reaction.Therefore, the content of gelatin need to control 1% and
Below.
Have detected the non-oxidizability of the sample obtained according to the method for embodiment 1-3 and reference examples 1-8, result such as table 1
Shown in.
The non-oxidizability of table 1 sample
DPPH method, ABTS method and ferrum reduction force method are usually used in the in vitro anti-oxidation evaluation of antioxidant content, DPPH and
ABTS free radical scavenging activity is the highest, and the light absorption value of ferrum reducing power is the highest, represents that the non-oxidizability of sample is the strongest.As shown in Table 1, press
Albumen-catechin free radical grafting the thing prepared according to the method for the present invention, albumen and hydrogen peroxide, ascorbic acid,
Synergism between catechin, has higher non-oxidizability, hence it is evident that be better than matched group.
Although the present invention is open the most as above with preferred embodiment, but it is not limited to the present invention, any is familiar with this skill
The people of art, without departing from the spirit and scope of the present invention, can do various changes and modification, therefore the protection model of the present invention
Enclosing should be with being as the criterion that claims are defined.
Claims (10)
1. the preparation method of albumen-catechin free radical grafting thing, it is characterised in that described method includes: the most pre-
Process Ovum Gallus domesticus album, to remove the mucin in Ovum Gallus domesticus album, adds radical initiator in egg white solution, then adds catechin and is formed
Albumen-catechin free radical grafting thing, then sample solution is carried out dialysis treatment to guarantee unreacted free catechin
Remove the most completely, after last lyophilization, obtain sample.
Method the most according to claim 1, it is characterised in that the mass ratio of the albumen in described catechin and egg white solution
For 1:20～1:5.
Method the most according to claim 1, it is characterised in that final concentration of the 0.5 of described catechin～2g/L.
Method the most according to claim 1, it is characterised in that described radical initiator is hydrogen peroxide and Vitamin C
Acid.
Method the most according to claim 1, it is characterised in that final concentration of the 0.05 of described hydrogen peroxide～0.2mol/
L, final concentration of the 1.25 of ascorbic acid～10g/L.
Method the most according to claim 1, it is characterised in that described Ovum Gallus domesticus album pretreatment is: add 9 times of volumes in Ovum Gallus domesticus album
Water dilution, final albumen quality mark is 1%, regulate pH to about 5.0, stir 10-20min, natural sedimentation 1h, then mistake
Filter off except insoluble protein ingredient.
Method the most according to claim 1, it is characterised in that described radical initiator and the reaction condition of albumen
It is 25 DEG C, 2h；Described catechin is 25 DEG C with the reaction condition of albumen, 12～48h.
Method the most according to claim 1, it is characterised in that described dialysis treatment is to carry out under the conditions of 4 DEG C.
9. albumen-catechin free radical grafting the thing obtained according to the arbitrary described method of claim 1-8.
10. albumen described in claim 9-catechin free radical grafting thing application in terms of food.
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| CN106723050A (en) * | 2017-01-03 | 2017-05-31 | 浙江大学 | The covalent thing of epigallocatechin ovalbumin stabilizes fish oil preparation and preparation method |
| CN106820150A (en) * | 2017-01-03 | 2017-06-13 | 浙江大学 | The covalent thing of catechin ovalbumin stabilizes fish oil preparation and preparation method thereof |
| CN106858601A (en) * | 2017-01-03 | 2017-06-20 | 浙江大学 | The covalent thing of Epigallo-catechin gallate (EGCG) ovalbumin stabilizes fish oil preparation and preparation method |
| CN107296283A (en) * | 2017-06-07 | 2017-10-27 | 东北农业大学 | A kind of preparation method of soybean protein-anthocyanidin compound |
| CN108822184A (en) * | 2018-04-25 | 2018-11-16 | 浙江工业大学 | A kind of method that protein function is modified |
| CN111919991A (en) * | 2020-07-14 | 2020-11-13 | 江苏大学 | Albumen protein-plant polyphenol covalent complex and preparation method thereof |
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| CN113598312A (en) * | 2021-07-27 | 2021-11-05 | 江苏大学 | Low-glycemic-index nutritional staple food fine dried noodles and production method thereof |
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| CN106723050A (en) * | 2017-01-03 | 2017-05-31 | 浙江大学 | The covalent thing of epigallocatechin ovalbumin stabilizes fish oil preparation and preparation method |
| CN106820150A (en) * | 2017-01-03 | 2017-06-13 | 浙江大学 | The covalent thing of catechin ovalbumin stabilizes fish oil preparation and preparation method thereof |
| CN106858601A (en) * | 2017-01-03 | 2017-06-20 | 浙江大学 | The covalent thing of Epigallo-catechin gallate (EGCG) ovalbumin stabilizes fish oil preparation and preparation method |
| CN107296283A (en) * | 2017-06-07 | 2017-10-27 | 东北农业大学 | A kind of preparation method of soybean protein-anthocyanidin compound |
| CN108822184A (en) * | 2018-04-25 | 2018-11-16 | 浙江工业大学 | A kind of method that protein function is modified |
| CN111919991A (en) * | 2020-07-14 | 2020-11-13 | 江苏大学 | Albumen protein-plant polyphenol covalent complex and preparation method thereof |
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| CN113598312A (en) * | 2021-07-27 | 2021-11-05 | 江苏大学 | Low-glycemic-index nutritional staple food fine dried noodles and production method thereof |
| CN114702692A (en) * | 2022-04-01 | 2022-07-05 | 中国科学院青岛生物能源与过程研究所 | Method for improving stability of protein hydrogel |
| CN114702692B (en) * | 2022-04-01 | 2024-04-23 | 中国科学院青岛生物能源与过程研究所 | Method for improving stability of protein hydrogel |
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Application publication date: 20161207 |