CN115124630A - Chitosan derivative and preparation method and application thereof - Google Patents
Chitosan derivative and preparation method and application thereof Download PDFInfo
- Publication number
- CN115124630A CN115124630A CN202110181835.1A CN202110181835A CN115124630A CN 115124630 A CN115124630 A CN 115124630A CN 202110181835 A CN202110181835 A CN 202110181835A CN 115124630 A CN115124630 A CN 115124630A
- Authority
- CN
- China
- Prior art keywords
- chitosan
- derivative
- acid
- salt
- chitosan derivative
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 229920001661 Chitosan Polymers 0.000 title claims abstract description 201
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 229940030275 epigallocatechin gallate Drugs 0.000 claims abstract description 38
- WMBWREPUVVBILR-UHFFFAOYSA-N GCG Natural products C=1C(O)=C(O)C(O)=CC=1C1OC2=CC(O)=CC(O)=C2CC1OC(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-UHFFFAOYSA-N 0.000 claims abstract description 25
- OEIJRRGCTVHYTH-UHFFFAOYSA-N Favan-3-ol Chemical group OC1CC2=CC=CC=C2OC1C1=CC=CC=C1 OEIJRRGCTVHYTH-UHFFFAOYSA-N 0.000 claims abstract description 23
- WMBWREPUVVBILR-WIYYLYMNSA-N (-)-Epigallocatechin-3-o-gallate Chemical compound O([C@@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=C(O)C=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-WIYYLYMNSA-N 0.000 claims abstract description 21
- XMOCLSLCDHWDHP-IUODEOHRSA-N epi-Gallocatechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@H]2O)=CC(O)=C(O)C(O)=C1 XMOCLSLCDHWDHP-IUODEOHRSA-N 0.000 claims abstract description 15
- XMOCLSLCDHWDHP-UHFFFAOYSA-N L-Epigallocatechin Natural products OC1CC2=C(O)C=C(O)C=C2OC1C1=CC(O)=C(O)C(O)=C1 XMOCLSLCDHWDHP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229950001002 cianidanol Drugs 0.000 claims abstract description 9
- DZYNKLUGCOSVKS-UHFFFAOYSA-N epigallocatechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3cc(O)c(O)c(O)c3 DZYNKLUGCOSVKS-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229940026509 theaflavin Drugs 0.000 claims abstract description 8
- 239000004410 anthocyanin Substances 0.000 claims abstract description 7
- 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 6
- WMBWREPUVVBILR-GHTZIAJQSA-N (+)-gallocatechin gallate Chemical compound O([C@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=C(O)C=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-GHTZIAJQSA-N 0.000 claims abstract description 6
- LSHVYAFMTMFKBA-TZIWHRDSSA-N (-)-epicatechin-3-O-gallate Chemical compound O([C@@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=CC=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 LSHVYAFMTMFKBA-TZIWHRDSSA-N 0.000 claims abstract description 6
- LSHVYAFMTMFKBA-UHFFFAOYSA-N ECG Natural products C=1C=C(O)C(O)=CC=1C1OC2=CC(O)=CC(O)=C2CC1OC(=O)C1=CC(O)=C(O)C(O)=C1 LSHVYAFMTMFKBA-UHFFFAOYSA-N 0.000 claims abstract description 6
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 235000005487 catechin Nutrition 0.000 claims abstract description 6
- LVJJFMLUMNSUFN-UHFFFAOYSA-N gallocatechin gallate Natural products C1=C(O)C=C2OC(C=3C=C(O)C(O)=CC=3)C(O)CC2=C1OC(=O)C1=CC(O)=C(O)C(O)=C1 LVJJFMLUMNSUFN-UHFFFAOYSA-N 0.000 claims abstract description 6
- PFTAWBLQPZVEMU-ZFWWWQNUSA-N (+)-epicatechin Natural products C1([C@@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-ZFWWWQNUSA-N 0.000 claims abstract description 5
- PFTAWBLQPZVEMU-UKRRQHHQSA-N (-)-epicatechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-UKRRQHHQSA-N 0.000 claims abstract description 5
- IPMYMEWFZKHGAX-UHFFFAOYSA-N Isotheaflavin Natural products OC1CC2=C(O)C=C(O)C=C2OC1C(C1=C2)=CC(O)=C(O)C1=C(O)C(=O)C=C2C1C(O)CC2=C(O)C=C(O)C=C2O1 IPMYMEWFZKHGAX-UHFFFAOYSA-N 0.000 claims abstract description 5
- UXRMWRBWCAGDQB-UHFFFAOYSA-N Theaflavin Natural products C1=CC(C2C(CC3=C(O)C=C(O)C=C3O2)O)=C(O)C(=O)C2=C1C(C1OC3=CC(O)=CC(O)=C3CC1O)=CC(O)=C2O UXRMWRBWCAGDQB-UHFFFAOYSA-N 0.000 claims abstract description 5
- 235000012734 epicatechin Nutrition 0.000 claims abstract description 5
- LPTRNLNOHUVQMS-UHFFFAOYSA-N epicatechin Natural products Cc1cc(O)cc2OC(C(O)Cc12)c1ccc(O)c(O)c1 LPTRNLNOHUVQMS-UHFFFAOYSA-N 0.000 claims abstract description 5
- IPMYMEWFZKHGAX-ZKSIBHASSA-N theaflavin Chemical compound C1=C2C([C@H]3OC4=CC(O)=CC(O)=C4C[C@H]3O)=CC(O)=C(O)C2=C(O)C(=O)C=C1[C@@H]1[C@H](O)CC2=C(O)C=C(O)C=C2O1 IPMYMEWFZKHGAX-ZKSIBHASSA-N 0.000 claims abstract description 5
- 235000014620 theaflavin Nutrition 0.000 claims abstract description 5
- 229930002877 anthocyanin Natural products 0.000 claims abstract description 4
- 235000010208 anthocyanin Nutrition 0.000 claims abstract description 4
- 150000004636 anthocyanins Chemical class 0.000 claims abstract description 4
- XMOCLSLCDHWDHP-SWLSCSKDSA-N (+)-Epigallocatechin Natural products C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC(O)=C(O)C(O)=C1 XMOCLSLCDHWDHP-SWLSCSKDSA-N 0.000 claims abstract description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 64
- 239000002253 acid Substances 0.000 claims description 38
- 238000010438 heat treatment Methods 0.000 claims description 33
- 238000000034 method Methods 0.000 claims description 22
- 235000011054 acetic acid Nutrition 0.000 claims description 21
- 150000003839 salts Chemical class 0.000 claims description 21
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 18
- 238000002156 mixing Methods 0.000 claims description 17
- 238000003756 stirring Methods 0.000 claims description 17
- 230000006196 deacetylation Effects 0.000 claims description 16
- 238000003381 deacetylation reaction Methods 0.000 claims description 16
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 15
- 229930182497 flavan-3-ol Natural products 0.000 claims description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- 239000012535 impurity Substances 0.000 claims description 11
- 238000006243 chemical reaction Methods 0.000 claims description 10
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 10
- 239000004310 lactic acid Substances 0.000 claims description 9
- 235000014655 lactic acid Nutrition 0.000 claims description 9
- 235000013305 food Nutrition 0.000 claims description 8
- 239000002537 cosmetic Substances 0.000 claims description 6
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 5
- 238000000576 coating method Methods 0.000 claims description 5
- 235000019253 formic acid Nutrition 0.000 claims description 5
- 230000035484 reaction time Effects 0.000 claims description 5
- 239000011248 coating agent Substances 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 235000015165 citric acid Nutrition 0.000 claims description 2
- 235000011167 hydrochloric acid Nutrition 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 36
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 30
- 230000003647 oxidation Effects 0.000 abstract description 12
- 238000007254 oxidation reaction Methods 0.000 abstract description 12
- 239000000463 material Substances 0.000 abstract description 2
- 230000001046 anti-mould Effects 0.000 abstract 1
- 239000002546 antimould Substances 0.000 abstract 1
- 238000004108 freeze drying Methods 0.000 description 39
- 238000000227 grinding Methods 0.000 description 39
- 239000000843 powder Substances 0.000 description 36
- 238000000502 dialysis Methods 0.000 description 19
- 239000000203 mixture Substances 0.000 description 19
- 238000005303 weighing Methods 0.000 description 19
- 238000000746 purification Methods 0.000 description 16
- 229910052739 hydrogen Inorganic materials 0.000 description 12
- 239000001257 hydrogen Substances 0.000 description 12
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 10
- 238000005481 NMR spectroscopy Methods 0.000 description 10
- 238000012360 testing method Methods 0.000 description 10
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 238000004611 spectroscopical analysis Methods 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 description 4
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 235000015278 beef Nutrition 0.000 description 4
- 230000009881 electrostatic interaction Effects 0.000 description 4
- -1 hydrogen ions Chemical class 0.000 description 4
- 239000004302 potassium sorbate Substances 0.000 description 4
- 235000010241 potassium sorbate Nutrition 0.000 description 4
- 229940069338 potassium sorbate Drugs 0.000 description 4
- 229940079593 drug Drugs 0.000 description 3
- CHHHXKFHOYLYRE-STWYSWDKSA-M potassium sorbate Chemical group [K+].C\C=C\C=C\C([O-])=O CHHHXKFHOYLYRE-STWYSWDKSA-M 0.000 description 3
- 238000001228 spectrum Methods 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
- 229920002101 Chitin Polymers 0.000 description 2
- 239000005714 Chitosan hydrochloride Substances 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- 230000002292 Radical scavenging effect Effects 0.000 description 2
- 125000000129 anionic group Chemical group 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 239000004599 antimicrobial Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 239000007806 chemical reaction intermediate Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 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 2
- 238000009920 food preservation Methods 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 230000002335 preservative effect Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000002834 transmittance Methods 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 241000228245 Aspergillus niger Species 0.000 description 1
- 241000193755 Bacillus cereus Species 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- 241000238557 Decapoda Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 108010029541 Laccase Proteins 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 241000555688 Malassezia furfur Species 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- 241000122123 Penicillium italicum Species 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 1
- MSWZFWKMSRAUBD-QZABAPFNSA-N beta-D-glucosamine Chemical compound N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-QZABAPFNSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229940095731 candida albicans Drugs 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 235000015223 cooked beef Nutrition 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
Images
Classifications
-
- 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
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
- C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the field of chitosan materials, and discloses a chitosan derivative, and a preparation method and application thereof. The chitosan derivative comprises flavan-3-ol structure and chitosan structure; the flavan-3-ol structure is connected with the chitosan structure through an ionic bond; the flavan-3-ol structure is at least one selected from catechin, epicatechin, gallocatechin, epigallocatechin, epicatechin gallate, gallocatechin gallate, epigallocatechin gallate, anthocyanin and theaflavin. The chitosan derivative has good antibacterial property and oxidation resistance, particularly good high temperature stability, for example, after being treated at a temperature of more than 120 ℃, the chitosan derivative still has good antibacterial property, and even the anti-mould property is further improved. In addition, the chitosan derivative has a solubility of not less than 1g/100g of water at 25 ℃.
Description
Technical Field
The invention belongs to the field of chitosan materials, and particularly relates to a chitosan derivative and a preparation method and application thereof.
Background
Chitin is a naturally occurring polymer formed by β -1,4 glycosidic linkages to N-acetyl-D-glucosamine and D-glucosamine. It can be processed to achieve partial or complete removal of acetyl groups and to obtain a polymer known as chitosan. Most commercial chitosan is obtained by first extracting chitin from fishery by-products (e.g., shrimp or crab shells) and then deacylating with alkali or acid.
Chitosan is known to have various biological properties, including its antibacterial activity, and thus can be applied to food or pharmaceutical industries. Chitosan is a food additive that has been approved in the national standard GB2760-2014, with specific functions being thickeners and coating agents. However, the application of chitosan has the following problems: first, it is poorly soluble in water; second, chitosan as an antimicrobial agent requires higher concentrations to be added for antimicrobial effect, and is more costly and less effective to apply than other antimicrobial agents. Therefore, in the specific application process of chitosan, the chitosan needs to be improved to meet the requirement of practical production.
The prior art introduces a preparation method of chitosan derivatives to prepare chitosan-polyphenol conjugates, and the antibacterial property, the oxidation resistance and the water solubility of the chitosan-polyphenol conjugates are improved compared with those of chitosan. However, the chitosan and the polyphenol conjugate are linked through covalent bonds, the preparation process needs to introduce catalytic enzyme (such as laccase) or introduce non-food-grade chemical reagent (such as hydrogen peroxide), the preparation process is relatively complex and high in cost, and the antibacterial property, the antioxidant property or the stability are not good. This has a certain barrier effect to the application of chitosan.
Therefore, there is a need to provide a novel chitosan derivative which not only has good antibacterial and antioxidant properties, but also has good stability, and facilitates the application of the chitosan derivative.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art described above. Therefore, the invention provides a chitosan derivative and a preparation method and application thereof, wherein the chitosan derivative has good antibacterial property and oxidation resistance, and particularly has good high-temperature stability, for example, after being treated at a temperature of more than 120 ℃, the chitosan derivative still has good antibacterial property, and even the antibacterial property is further improved. In addition, the water solubility of the chitosan derivative is remarkably improved compared with that of chitosan.
The invention conception of the invention is as follows: the chitosan derivative is a salt formed by electrostatic interaction between hydroxyl on flavan-3-ol and chitosan side chain amino. The preparation process of the chitosan derivative needs chitosan-acid salt as an important reaction intermediate product, then the anionic group formed by acid in the chitosan-acid salt is replaced by the flavan-3-alcohol group to generate chitosan-flavan-3-alkoxide, and the hydroxyl in the flavan-3-alcohol reacts with the amino in the chitosan to form an ionic bond. In the chitosan acid salt derivative in the prior art, the reacted group is a covalent bond conjugate derivative generated by the reaction of hydrogen ions in organic acid and hydroxyl groups of chitosan side chains.
In a first aspect of the invention, there is provided a chitosan derivative.
Specifically, a chitosan derivative comprises a flavan-3-ol structure and a chitosan structure; the flavan-3-ol structure is connected with the chitosan structure through ionic bonds.
The chitosan derivative is a salt, the chitosan structure serves as a positive ion, the flavan-3-ol structure serves as an anion, and hydroxyl in the flavan-3-ol and amino in the chitosan are subjected to electrostatic interaction to form the salt.
Preferably, the flavan-3-ol structure is at least one selected from catechin, epicatechin, gallocatechin, epigallocatechin, epicatechin gallate, gallocatechin gallate, epigallocatechin gallate, anthocyanin and theaflavin.
Preferably, the degree of deacetylation of the chitosan structure is 50-100%; further preferably, the degree of deacetylation of the chitosan structure is 80-99%.
Preferably, the chitosan derivative has a molecular weight of 1kDa to 5000 kDa; further preferably, the chitosan derivative has a molecular weight of 1kDa to 2500 kDa; more preferably from 120kDa to 1000 kDa.
Preferably, the chitosan derivative is at least one selected from chitosan-catechin salt, chitosan-epicatechin salt, chitosan-epigallocatechin salt, chitosan-epicatechin gallate salt, chitosan-epigallocatechin gallate salt, chitosan-gallocatechin gallate salt and chitosan-theaflavin salt.
Preferably, the structural formula of the chitosan-epigallocatechin gallate is shown in the specification
Wherein the value of n is 1-10000. The epigallocatechin gallate and the chitosan are connected through ionic bonds (the chitosan-epigallocatechin gallate can also be formed by electrostatic interaction between any hydroxyl on the epigallocatechin gallate structure and amino in the chitosan structure to form ionic bonds).
The second aspect of the present invention provides a method for preparing a chitosan derivative.
Specifically, the preparation method of the chitosan derivative comprises the following steps:
mixing chitosan with acid solution, heating to obtain chitosan-acid salt, adding flavan-3-ol, stirring, and reacting to obtain the chitosan derivative.
Preferably, the chitosan has a molecular weight of 1kDa to 4000 kDa; further preferably, the chitosan has a molecular weight of 1kDa to 1500 kDa; more preferably from 120kDa to 840 kDa.
Preferably, the chitosan has a deacetylation degree of 50-100%; further preferably, the chitosan has a degree of deacetylation of 80 to 99%.
Preferably, the acid solution is selected from a solution of hydrochloric acid, formic acid, lactic acid, citric acid or acetic acid.
Preferably, the volume concentration of the acid liquor is 0.5-10%; further preferably, the volume concentration of the acid solution is 2-8% (the volume concentration of the acid solution refers to the volume concentration of the acid in the mixture formed after the acid solution is mixed with the chitosan).
Preferably, the mass volume ratio of the chitosan to the acid liquor is (5-20) g (5-30) mL; further preferably, the mass volume ratio of the chitosan to the acid liquor is (8-15) g (10-25) mL.
Preferably, the pH of a mixture formed by mixing the chitosan and the acid solution is 1-6; further preferably, the pH is 2 to 5.
Preferably, the heating temperature is 30-300 ℃; further preferably, the heating temperature is 50 to 100 ℃.
Preferably, the heating time is 10 to 500 minutes; further preferably, the heating time is 20 to 200 minutes.
Preferably, the mass ratio of the added flavan-3-ol to the chitosan-acid salt is (0.1-5) to 1; more preferably, the mass ratio of the added flavan-3-ol to the chitosan-acid salt is (0.5-4): 1.
Preferably, the temperature of the reaction is 30-300 ℃; further preferably, the temperature of the reaction is 50 to 100 ℃.
Preferably, the reaction time is 10 to 500 minutes; further preferably, the reaction time is 20 to 200 minutes. The addition amount of flavan-3-ol, the proper reaction temperature and the proper reaction time of the chitosan-acid salt enable the prepared chitosan derivative to have better antibacterial property, and particularly, the prepared chitosan derivative can still keep good antibacterial property after being treated at high temperature (such as 120 ℃).
Preferably, a method for preparing a chitosan derivative comprises the following steps:
mixing chitosan with acid solution, heating, removing impurities for the first time to obtain chitosan-acid salt, adding flavan-3-ol, stirring, reacting, and removing impurities for the second time to obtain the chitosan derivative.
Preferably, the specific process of the first impurity removal is as follows: the mixture obtained after heating was dried and then purified by dialysis (commercially available cellulose filter membranes, pore size adjusted according to the molecular weight requirement, and purified by dialysis with ultrapure water at room temperature for 3 to 7 days) to remove the excess unreacted acid. Helps to reduce impurities in the chitosan-acid salt.
Preferably, the second impurity removal specifically comprises the following steps: the mixture obtained after the reaction is dried and then purified by dialysis to remove the excess unreacted flavan-3-ol. Is helpful for reducing impurities in the chitosan derivative, thereby improving the antibacterial property and the oxidation resistance of the chitosan derivative.
More preferably, a method for preparing a chitosan derivative comprises the following steps:
mixing chitosan with acid liquor, heating, drying, removing impurities for the first time to obtain chitosan-acid salt, mixing the chitosan-acid salt with a solvent, adding flavan-3-ol, stirring, reacting, drying, and removing impurities for the second time to obtain the chitosan derivative.
Preferably, drying and grinding are further carried out after the second impurity removal, so as to obtain the powdery chitosan derivative. Is helpful for the use of the chitosan derivative.
The drying can be air drying, freeze drying, spray drying or heat drying.
The third aspect of the present invention provides a use of a chitosan derivative.
The chitosan derivative is applied to preparation of medicines, foods, cosmetics or coatings.
Preferably, the coating is an antibacterial coating.
Preferably, the food or cosmetic is a food or cosmetic having good antibacterial and antiseptic effects.
Compared with the prior art, the invention has the following beneficial effects:
(1) the chitosan derivative is a salt formed by electrostatic interaction between hydroxyl on flavan-3-ol and chitosan side chain amino. The preparation process of the chitosan derivative needs chitosan-acid salt as an important reaction intermediate product, then the anionic group formed by acid in the chitosan-acid salt is replaced by the flavan-3-alcohol group to generate chitosan-flavan-3-alkoxide, and the hydroxyl in the flavan-3-alcohol reacts with the amino in the chitosan to form an ionic bond. In the chitosan acid salt derivative in the prior art, the reacted group is a covalent bond conjugate derivative generated by the reaction of hydrogen ions in organic acid and hydroxyl groups of chitosan side chains. Therefore, the chitosan derivative has better water solubility (at 25 ℃, the solubility is not less than 1g/100g of water, for example, 1g/100g of water to 12g/100g of water), good antibacterial property and oxidation resistance, and particularly good high-temperature resistance stability, for example, after being treated at the temperature of more than 120 ℃, the chitosan derivative still has good antibacterial property, and even the mould property is further improved.
(2) The chitosan derivative prepared by the invention has good high temperature stability, and good antibacterial property and oxidation resistance, so the chitosan derivative can be widely applied to medicines, foods and cosmetics.
Drawings
FIG. 1 is a Fourier infrared spectrum of chitosan-epigallocatechin gallate prepared in example 1.
Detailed Description
In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples are given for illustration. It should be noted that the following examples are not intended to limit the scope of the claimed invention.
The starting materials, reagents or apparatuses used in the following examples are conventionally commercially available or can be obtained by conventionally known methods, unless otherwise specified.
Example 1: preparation of chitosan derivative (chitosan-epigallocatechin gallate)
A method for preparing chitosan derivative (chitosan-epigallocatechin gallate) comprises the following steps:
selecting chitosan with the molecular weight of 280kDa and the deacetylation degree of 88% (subjected to nuclear magnetic resonance hydrogen spectrometry), weighing 10g of the chitosan, dissolving the chitosan in 1L of water, adding acetic acid (the volume concentration of the acetic acid is 2%, namely the volume concentration of the acetic acid in a mixture formed by the chitosan, the water and the acetic acid is 2%), mixing to form a mixture, wherein the pH of the mixture is 3.5, heating the mixture at 50 ℃ for 1 hour, performing freeze-drying treatment for 24 hours after the heating is finished, grinding the mixture to obtain powder, performing dialysis purification treatment on the powder to remove excessive unreacted acetic acid, and performing freeze-drying and grinding treatment again to obtain chitosan-acetate powder;
weighing 10g of chitosan-acetate powder, dissolving in 1L of water, adding 10g of epigallocatechin gallate, carrying out stirring reaction at 70 ℃ for 1.5 hours under a closed condition (the reaction is carried out under the closed condition to avoid volatilization of acetic acid), carrying out freeze-drying and grinding treatment for 24 hours, carrying out dialysis purification to remove the epigallocatechin gallate which is not completely reacted, and carrying out freeze-drying and grinding treatment to obtain the chitosan derivative (chitosan-epigallocatechin gallate).
FIG. 1 is a Fourier infrared spectrum of the chitosan-epigallocatechin gallate obtained in example 1 (in FIG. 1, the ordinate represents "Transmittance", the abscissa represents "Wave number", and the abscissa represents "cm" in units of "Transmittance", and "Wave number", respectively -1 "; CHIT represents chitosan, CHIT-AC represents chitosan-acetate, CHIT-FLAV represents chitosan-epigallocatechin gallate, and FLAV represents epigallocatechin gallate). As can be seen from FIG. 1, 1556cm in CHIT-AC -1 At peak, is carboxylate anion COO - ，1408cm -1 Peak, tensile mode of carboxylate anion, FLAV and CHIT-FLAV, 1693cm -1 And 1606cm -1 Peak at 1230cm in FLAV and CHIT-FLAV, representing C ═ C double bond in epigallocatechin gallate benzene ring -1 The peak indicates the C-O bond in the hydroxyl group of epigallocatechin gallate.
The intermediate product chitosan-acetate and the product chitosan-epigallocatechin gallate obtained in this example 1 were subjected to structural analysis by fourier infrared spectroscopy, and the spectra thereof are shown in fig. 1. From FIG. 1, it can be seen that chitosan-acetate is an important intermediate in the synthesis step, and then the acetate group is replaced by epigallocatechin gallate group to form chitosan-epigallocatechin gallate with different structural characteristics from chitosan, epigallocatechin gallate and chitosan acetate.
The structural formula of chitosan is:
(since the chitosan molecular weight chosen is 280kDa, n is also defined here).
The structural formula of the chitosan-acetate is as follows:
(since the chitosan molecular weight chosen is 280kDa, n is also defined here).
The structural formula of the chitosan-epigallocatechin gallate is as follows:
(since the chitosan molecular weight chosen is 280kDa, n is also defined here).
Example 2: preparation of chitosan derivative (chitosan-catechin salt)
A method for preparing chitosan derivative (chitosan-catechin salt) comprises the following steps:
selecting chitosan with molecular weight of 100kDa and deacetylation degree of 80% (by nuclear magnetic resonance hydrogen spectrometry), weighing 8g of the chitosan, dissolving in 1L of water, adding hydrochloric acid (the volume concentration of the hydrochloric acid is 3%), mixing to form a mixture, wherein the pH of the mixture is 3.0, heating at 40 ℃ for 40 minutes, performing 28-hour freeze-drying treatment after heating is completed, grinding to obtain powder, performing dialysis purification treatment on the powder, removing excessive unreacted hydrochloric acid, and performing freeze-drying and grinding again to obtain chitosan-hydrochloride powder;
weighing 10g of chitosan-hydrochloride powder, dissolving in 1L of water, adding 15g of catechin, stirring and reacting for 2 hours at 75 ℃ under a closed condition, then carrying out freeze-drying and grinding treatment for 24 hours, then carrying out dialysis purification, removing catechin which is not completely reacted, and then carrying out freeze-drying and grinding treatment to obtain the chitosan derivative (chitosan-catechin salt).
Example 3: preparation of chitosan derivative (chitosan-epicatechin salt)
A method for preparing chitosan derivative (chitosan-epicatechin salt) comprises the following steps:
selecting chitosan with the molecular weight of 1000kDa and the deacetylation degree of 70% (subjected to nuclear magnetic resonance hydrogen spectrometry), weighing 12g of the chitosan, dissolving the chitosan in 1L of water, adding acetic acid (the volume concentration of the acetic acid is 4%), mixing to form a mixture, heating at 60 ℃ for 50 minutes, carrying out freeze-drying treatment for 24 hours after heating is finished, grinding to obtain powder, carrying out dialysis purification treatment on the powder, removing excessive unreacted acetic acid, and then carrying out freeze-drying and grinding treatment again to obtain chitosan-acetate powder;
weighing 10g of chitosan-acetate powder, dissolving in 1L of water, adding 11g of epicatechin, reacting under a closed condition at 70 ℃ for 2 hours under stirring, carrying out freeze-drying and grinding treatment for 24 hours, then carrying out dialysis purification, removing the epicatechin which is not completely reacted, and then carrying out freeze-drying and grinding treatment to obtain the chitosan derivative (chitosan-epicatechin salt).
Example 4: preparation of chitosan derivative (chitosan-epigallocatechin salt)
A method for preparing chitosan derivative (chitosan-epigallocatechin salt) comprises the following steps:
selecting chitosan with the molecular weight of 1100kDa and the deacetylation degree of 75% (subjected to nuclear magnetic resonance hydrogen spectrometry), weighing 10g of the chitosan, dissolving the chitosan in 1L of water, adding acetic acid (the volume concentration of the acetic acid is 2.5%), mixing to form a mixture, heating at 70 ℃ for 70 minutes, carrying out freeze-drying treatment for 24 hours after heating is completed, grinding to obtain powder, carrying out dialysis purification treatment on the powder, removing excessive unreacted acetic acid, and then carrying out freeze-drying and grinding treatment again to obtain chitosan-acetate powder;
weighing 10g of chitosan-acetate powder, dissolving in 1L of water, adding 18g of epigallocatechin, stirring and reacting for 1 hour at 80 ℃ under a closed condition, freeze-drying and grinding for 24 hours, then dialyzing and purifying to remove the epigallocatechin which is not completely reacted, and freeze-drying and grinding to obtain the chitosan derivative (chitosan-epigallocatechin salt).
Example 5: preparation of chitosan derivative (chitosan-epicatechin gallate)
A method for preparing chitosan derivative (chitosan-epicatechin gallate) comprises the following steps:
selecting chitosan (subjected to nuclear magnetic resonance hydrogen spectrum verification) with the molecular weight of 1200kDa and the deacetylation degree of 85 percent, weighing 10g of the chitosan, dissolving the chitosan in 1L of water, adding formic acid (the volume concentration of the formic acid is 2 percent), mixing to form a mixture, heating at 75 ℃ for 75 minutes, carrying out freeze-drying treatment for 24 hours after heating is finished, grinding to obtain powder, carrying out dialysis purification treatment on the powder, removing redundant unreacted formic acid, and then carrying out freeze-drying and grinding treatment again to obtain chitosan-formate powder;
weighing 10g of chitosan-formate powder, dissolving in 1L of water, adding 20g of epicatechin gallate, reacting under sealed conditions at 85 ℃ for 1.5 hours under stirring, then carrying out freeze-drying and grinding treatment for 24 hours, then carrying out dialysis purification, removing the epicatechin gallate which is not completely reacted, and then carrying out freeze-drying and grinding treatment to obtain the chitosan derivative (chitosan-epicatechin gallate).
Example 6: preparation of chitosan derivative (chitosan-gallocatechin gallate)
A method for preparing chitosan derivative (chitosan-gallocatechin gallate) comprises the following steps:
selecting chitosan (subjected to nuclear magnetic resonance hydrogen spectroscopy verification) with the molecular weight of 1500kDa and the deacetylation degree of 88 percent, weighing 10g of the chitosan, dissolving the chitosan in 1L of water, adding lactic acid (the volume concentration of the lactic acid is 2 percent), mixing to form a mixture, heating at 75 ℃ for 75 minutes, performing freeze-drying treatment for 24 hours after heating is completed, grinding to obtain powder, performing dialysis purification treatment on the powder, removing redundant unreacted lactic acid, and performing freeze-drying and grinding treatment again to obtain chitosan-lactate powder;
weighing 10g of chitosan-lactate powder, dissolving in 1L of water, adding 22g of gallocatechin gallate, reacting for 2 hours under sealed conditions at 90 ℃, then carrying out freeze-drying and grinding treatment for 24 hours, then carrying out dialysis purification, removing the gallocatechin gallate which is not completely reacted, and then carrying out freeze-drying and grinding treatment to obtain the chitosan derivative (chitosan-gallocatechin gallate).
Example 7: preparation of chitosan derivative (chitosan-theaflavin salt)
A method for preparing chitosan derivative (chitosan-theaflavin salt) comprises the following steps:
selecting chitosan with the molecular weight of 100kDa and the deacetylation degree of 88% (subjected to nuclear magnetic resonance hydrogen spectrometry), weighing 10g of the chitosan, dissolving the chitosan in 1L of water, adding citric acid (the volume concentration of lactic acid is 2.5%), mixing to form a mixture, heating at 80 ℃ for 70 minutes, carrying out freeze-drying treatment for 24 hours after heating is completed, grinding to obtain powder, carrying out dialysis purification treatment on the powder, removing excessive unreacted citric acid, and then carrying out freeze-drying and grinding treatment again to obtain chitosan-citrate powder;
weighing 10g of chitosan-citrate powder to be dissolved in 1L of water, adding 13g of theaflavin, stirring and reacting for 2 hours at 90 ℃ under a closed condition, then carrying out freeze-drying and grinding treatment for 24 hours, subsequently carrying out dialysis purification to remove the unreacted theaflavin, and then carrying out freeze-drying and grinding treatment to obtain the chitosan derivative (chitosan-theaflavin salt).
Example 8: preparation of chitosan derivative (chitosan-epigallocatechin gallate)
A method for preparing chitosan derivative (chitosan-epigallocatechin gallate) comprises the following steps:
selecting chitosan with the molecular weight of 180kDa and the deacetylation degree of 88% (subjected to nuclear magnetic resonance hydrogen spectrometry), weighing 10g of the chitosan, dissolving the chitosan in 1L of water, adding acetic acid (the volume concentration of the acetic acid is 2.5%), mixing to form a mixture, heating at 80 ℃ for 1 hour, carrying out freeze-drying treatment for 24 hours after heating is completed, grinding to obtain powder, carrying out dialysis purification treatment on the powder, removing excessive unreacted acetic acid, and then carrying out freeze-drying and grinding treatment again to obtain chitosan-acetate powder;
weighing 10g of chitosan-acetate powder, dissolving in 1L of water, adding 6g of epigallocatechin gallate, reacting under sealed conditions at 70 ℃ for 1.5 hours under stirring, freeze-drying and grinding for 24 hours, then dialyzing and purifying, and freeze-drying and grinding to obtain the chitosan derivative (chitosan-epigallocatechin gallate).
Example 9: preparation of chitosan derivative (chitosan-anthocyanin salt)
A method for preparing chitosan derivative (chitosan-anthocyanin salt) comprises the following steps:
selecting chitosan with molecular weight of 200kDa and deacetylation degree of 88% (by nuclear magnetic resonance hydrogen spectrum check), weighing 10g of the chitosan, dissolving in 1L of water, adding lactic acid (the volume concentration of lactic acid is 2.5%), mixing to form a mixture, heating at 85 ℃ for 2 hours, after heating, carrying out freeze-drying treatment for 24 hours, grinding to obtain powder, carrying out dialysis purification treatment on the powder, removing excessive unreacted lactic acid, and then carrying out freeze-drying and grinding treatment again to obtain chitosan-lactate powder;
weighing 10g of chitosan-lactate powder, dissolving in 1L of water, adding 15g of anthocyanin, reacting for 2 hours under a closed condition at 70 ℃ under stirring, carrying out freeze-drying and grinding treatment for 24 hours, carrying out dialysis purification, and carrying out freeze-drying and grinding treatment to obtain the chitosan derivative (chitosan-anthocyanin salt).
Example 10
The only difference in example 10 compared to example 1 is that the temperature for the reaction of chitosan-acetate with epigallocatechin gallate in example 10 was 110 ℃ and the reaction time was 1.5 hours, and the rest of the procedure was the same as in example 1.
Comparative example 1 (preparation of chitosan-epigallocatechin gallate conjugate product of the prior art)
Selecting chitosan with molecular weight of 280kDa and deacetylation degree of 88% (subjected to nuclear magnetic resonance hydrogen spectrometry), weighing 10g of the chitosan, dissolving in 1L of water, adding acetic acid (the volume concentration of acetic acid is 2%, namely the volume concentration of acetic acid in the mixture formed by chitosan, water and acetic acid is 2%), mixing, stirring overnight to obtain chitosan solution, taking 100mL of the chitosan solution, adding 1mol/L hydrochloric acid to adjust the pH to 3.5, adding 1mL of 0.5mol/L hydrogen peroxide and 0.025g ascorbic acid, stirring at 40 ℃ for 1 hour, adding 1g epigallocatechin gallate, reacting at 40 deg.C under stirring for 12 hr, then freeze-drying and grinding for 24 hours, then dialyzing and purifying to remove the epigallocatechin gallate which is not completely reacted, then freeze-drying and grinding to obtain the chitosan-epigallocatechin gallate conjugate product.
Product effectiveness testing
1. Test of antibacterial Effect
The chitosan derivatives prepared in examples 1-7, 10 and 1 were tested for their Minimal Inhibitory Concentrations (MIC) against bacteria (including gram-positive bacteria including Staphylococcus aureus, Bacillus cereus and Lactobacillus plantarum, and gram-negative bacteria including Escherichia coli and Pseudomonas aeruginosa), yeasts (Malassezia furfur and Candida albicans), molds (Aspergillus niger and Penicillium italicum) (sample inoculation concentrations in ppm, and concentration gradients of 4000ppm, 2000ppm, 1000ppm and 500 ppm).
The cultured system was a commercially available nutrient broth (supplied by Kyork, Guangdong, Inc., model number 022010), the pH of the cultured system was 6, the culture conditions of bacteria were 7 days at a temperature of 36 ℃ and the culture conditions of yeast and mold were 7 days at a temperature of 28 ℃, and the results are shown in Table 1.
Table 1: antibacterial Effect (data in Table 1 represent MIC in ppm)
Remarking: in table 1 "/" indicates no bacteriostatic effect.
As can be seen from Table 1, the chitosan derivatives obtained in examples 1-7 and example 10 of the present invention have better antibacterial effects than comparative example 1. On the other hand, the chitosan-epigallocatechin gallate conjugate product prepared in comparative example 1 has no antibacterial effect on mold. As can be seen from the data of examples 1 and 10, the temperature at which the chitosan-acetate and epigallocatechin gallate are reacted has a certain effect on the antibacterial effect of the prepared chitosan derivative. The antibacterial effect of the chitosan derivative prepared in the remaining examples was similar to that of example 1.
2. Stability test
The chitosan derivative prepared in example 1 was used to test the antibacterial effect after being treated at different temperature conditions, and further used to test the stability of the chitosan derivative prepared in example 1. The system and the culture conditions were the same as above (the culture conditions for bacteria were 36 ℃ C. for 7 days, and the culture conditions for yeast and mold were 28 ℃ C. for 7 days). Different temperature conditions are specifically divided into: normal temperature of 25 ℃ for 1 hour, high pressure wet heat treatment (101KPa, 121 ℃, 15 minutes), normal pressure water bath at 121 ℃ for 1 hour, and normal pressure oil bath at 180 ℃ for 1 hour. The antimicrobial results are shown in table 2.
Table 2: antibacterial Effect (data in Table 2 represent MIC in ppm)
As can be seen from Table 2, after the chitosan derivative prepared in example 1 is treated under the conditions of high pressure, wet heat and humidity for 1 hour, normal pressure water bath at 121 ℃ for 1 hour, normal pressure oil bath at 180 ℃ for 1 hour and the like, the antibacterial effect on gram-positive bacteria, gram-negative bacteria and yeast is consistent with that at the normal temperature of 25 ℃, which indicates that the chitosan derivative prepared in example 1 of the present invention has good high temperature stability. In addition, the inventors have also found that the chitosan derivative obtained in example 1 has an unexpectedly enhanced antibacterial effect against mold after being subjected to the conditions of high-pressure moist heat treatment for 1 hour, normal-pressure water bath treatment for 1 hour at 121 ℃, and normal-pressure oil bath treatment for 1 hour at 180 ℃.
The chitosan derivatives obtained in the remaining examples had high temperature stability similar to that of the chitosan derivative obtained in example 1 above.
3. Oxidation resistance test
The chitosan derivatives obtained in examples 1 to 6 of the present invention were tested for their oxidation resistance (the strength of oxidation resistance was measured by DPPH radical scavenging rate, which is 1, 1-diphenyl-2-trinitrophenylhydrazine), and the results are shown in Table 3.
Table 3: effect of oxidation resistance
| Sample (I) | DPPH radical scavenging ratio (%) |
| Chitosan | 0 |
| Catechin | 54 |
| Example 2 | 50 |
| Epimetechin | 68 |
| Example 3 | 59 |
| Epicatechin gallate | 78 |
| Example 5 | 65 |
| Epigallocatechin | 73 |
| Example 4 | 61 |
| Epigallocatechin gallate | 84 |
| Example 1 | 72 |
| Gallocatechin gallate | 74 |
| Example 6 | 60 |
| Comparative example 1 | 73 |
As can be seen from Table 3, the chitosan derivatives prepared in examples 1-6 still have good oxidation resistance.
4. Application effect in food preservation and fresh-keeping
The chitosan derivative prepared in example 1 is taken and tested for application effect in food preservation and fresh keeping.
The experimental procedure was as follows:
(1) after the frozen beef is unfrozen, cutting the frozen beef into blocks, wherein each block is about 300-400 g;
(2) experimental groups, specifically 5 groups, which were blank group, chitosan group, epigallocatechin gallate group, chitosan derivative group prepared in example 1, and potassium sorbate group,
(3) preparing saline solution with the mass concentration of 2% for each group, respectively adding chitosan (the concentration of chitosan in the saline solution is 3000ppm), epigallocatechin gallate (the concentration of epigallocatechin gallate in the saline solution is 3000ppm), the chitosan derivative prepared in example 1 (the concentration of the chitosan derivative prepared in example 1 in the saline solution is 3000ppm), potassium sorbate (the concentration of potassium sorbate in the saline solution is 75ppm), recording the original pH value of the saline solution in the potassium sorbate group, adding potassium sorbate after adjusting the pH to 5.0 by adding citric acid, uniformly stirring to exert the antibacterial action of the potassium sorbate), heating in a pot, putting 2 beef blocks, stewing for about 1 hour by slow fire after boiling by strong fire, and preferably stewing the beef in soft and soft but not dispersing;
(4) cutting the cooked beef into small blocks of about 20g, filling 2 blocks into one bag, and directly sealing by heat, wherein each group is sealed by 14 bags;
(5) cold storing, testing colony count on day 0, taking 2 parallel samples for each test, and testing colony counts on days 1, 2, 3, 4 and 7;
(6) the test is repeated for two rounds, and the final result is based on the average value of two independent test data.
The results of monitoring the total number of colonies are shown in Table 4.
Table 4: total number of colonies (unit: log cfu/mL)
As can be seen from Table 4, after 7 days of food storage, the total number of colonies in the blank group had reached 7.96log cfu/mL and the total number of colonies in the potassium sorbate group was 4.12log cfu/mL, while the chitosan derivative group prepared in example 1 was maintained at 1.02log cfu/mL. As can be seen, the chitosan derivative prepared in example 1 has good preservative and fresh-keeping effects. The chitosan derivatives prepared in other examples also have similar preservative and fresh-keeping effects.
Addition of chitosan and epigallocatechin gallate alone to the above food products does not have good antibacterial properties.
In addition, the chitosan derivative prepared by the invention has good high-temperature stability, and good antibacterial property and oxidation resistance, so the chitosan derivative can be widely applied to medicines, cosmetics or coatings.
5. Dissolution effect
The chitosan derivative prepared in example 1 of the present invention had a solubility in water of 10g/100g water, a solubility of chitosan of less than 0.01g/100g water, and a solubility of the chitosan-epigallocatechin gallate conjugate product prepared in comparative example 1 of less than 0.01g/100g water at room temperature of 25 ℃.
Claims (15)
1. A chitosan derivative, comprising a flavan-3-ol structure and a chitosan structure; the flavan-3-ol structure is connected with the chitosan structure through an ionic bond.
2. The chitosan derivative of claim 1, wherein the flavan-3-ol structure is selected from at least one of catechin, epicatechin, gallocatechin, epigallocatechin, epicatechin gallate, gallocatechin gallate, epigallocatechin gallate, anthocyanin, or theaflavin.
3. Chitosan derivative according to claim 1, characterized in that the degree of deacetylation of the chitosan structure is between 50-100%.
4. The chitosan derivative according to claim 1, wherein the chitosan derivative has a molecular weight of 1kDa to 5000 kDa.
5. The chitosan derivative according to claim 1, wherein the chitosan derivative is at least one selected from the group consisting of chitosan-catechin salt, chitosan-epicatechin salt, chitosan-epigallocatechin salt, chitosan-epicatechin gallate salt, chitosan-epigallocatechin gallate salt, chitosan-gallocatechin gallate salt, and chitosan-theaflavin salt.
6. The chitosan derivative according to claim 5, wherein the chitosan-epigallocatechin gallate has a structural formula of
Wherein the value of n is 1-10000.
7. The method for producing a chitosan derivative according to any one of claims 1 to 6, comprising the steps of:
mixing chitosan with acid solution, heating to obtain chitosan-acid salt, adding flavan-3-ol, stirring, and reacting to obtain the chitosan derivative.
8. The method of claim 7, wherein the chitosan has a molecular weight of 1kDa to 4000 kDa.
9. The method according to claim 7, wherein the degree of deacetylation of chitosan is 50 to 100%.
10. The method according to claim 7, wherein the acid solution is selected from a solution of hydrochloric acid, formic acid, lactic acid, citric acid, or acetic acid.
11. The preparation method according to claim 7, wherein the acid solution has a volume concentration of 0.5-10%; the mass volume ratio of the chitosan to the acid liquor is (5-20) g (5-30) mL.
12. The method of claim 7, wherein the heating temperature is 30-300 ℃; the heating time is 10-500 minutes.
13. The method of claim 7, wherein the mass ratio of flavan-3-ol to chitosan-acid salt is (0.1-5): 1; the reaction temperature is 30-300 ℃; the reaction time is 10-500 minutes.
14. The method of claim 7, comprising the steps of:
mixing chitosan with acid solution, heating, removing impurities for the first time to obtain chitosan-acid salt, adding flavan-3-ol, stirring, reacting, and removing impurities for the second time to obtain the chitosan derivative.
15. Use of a chitosan derivative as defined in any of claims 1 to 6 in the preparation of a medicament, food, cosmetic or coating.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110181835.1A CN115124630B (en) | 2021-02-08 | 2021-02-08 | Chitosan derivative and preparation method and application thereof |
| PCT/CN2021/082319 WO2022165940A1 (en) | 2021-02-08 | 2021-03-23 | Chitosan derivative, preparation method therefor and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110181835.1A CN115124630B (en) | 2021-02-08 | 2021-02-08 | Chitosan derivative and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115124630A true CN115124630A (en) | 2022-09-30 |
| CN115124630B CN115124630B (en) | 2024-02-23 |
Family
ID=82740755
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110181835.1A Active CN115124630B (en) | 2021-02-08 | 2021-02-08 | Chitosan derivative and preparation method and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN115124630B (en) |
| WO (1) | WO2022165940A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116554704A (en) * | 2023-04-17 | 2023-08-08 | 深圳力越新材料有限公司 | Liquid crystal polymer material for 5G and preparation method thereof |
| CN116554363A (en) * | 2023-04-18 | 2023-08-08 | 珠海市自然之旅生物技术有限公司 | Torone-chitosan derivative and preparation method and application thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117143677A (en) * | 2023-10-14 | 2023-12-01 | 广州市爱家有方日用品有限公司 | Green and safe sweat-odor-removing laundry detergent and preparation method thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1486700A (en) * | 2002-09-30 | 2004-04-07 | 王洪栋 | Proanthocyanidin compound and its prepn |
| CN101974102A (en) * | 2010-10-28 | 2011-02-16 | 浙江工业大学 | Novel water soluble chitosan derivative and preparation and application thereof |
| CN106084088A (en) * | 2016-06-06 | 2016-11-09 | 武汉艾美博特医疗用品有限公司 | The Preparation method and use of chitosan gallic acid copolymer |
| CN106893004A (en) * | 2017-03-17 | 2017-06-27 | 广西大学 | A kind of preparation method of shitosan phenolic acid conjugates |
| CN109336081A (en) * | 2018-08-29 | 2019-02-15 | 北京化工大学常州先进材料研究院 | A kind of preparation method of the lithium ion battery spherical carbon cathode based on chitosan |
| CN111514097A (en) * | 2020-05-14 | 2020-08-11 | 燕山大学 | Preparation method of pH-responsive nano hydrogel of walnut shell polyphenol |
| CN111961263A (en) * | 2020-08-28 | 2020-11-20 | 浙江奚态生物科技有限公司 | Preparation method of antibacterial degradable food packaging film material |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007262400A (en) * | 2006-03-03 | 2007-10-11 | Hisashi Yoshioka | Polyphenol-containing plant extract- chitosan composite material and method for producing the same |
| US20080009411A1 (en) * | 2006-07-05 | 2008-01-10 | Hsien-Chih Lin | Method for Manufacturing Absorbent having Function of Suppressing Leakage of Formaldehyde |
| TWI439288B (en) * | 2012-10-05 | 2014-06-01 | Univ China Medical | Medicinal carriers and preparation method and uses thereof |
-
2021
- 2021-02-08 CN CN202110181835.1A patent/CN115124630B/en active Active
- 2021-03-23 WO PCT/CN2021/082319 patent/WO2022165940A1/en active Application Filing
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1486700A (en) * | 2002-09-30 | 2004-04-07 | 王洪栋 | Proanthocyanidin compound and its prepn |
| CN101974102A (en) * | 2010-10-28 | 2011-02-16 | 浙江工业大学 | Novel water soluble chitosan derivative and preparation and application thereof |
| CN106084088A (en) * | 2016-06-06 | 2016-11-09 | 武汉艾美博特医疗用品有限公司 | The Preparation method and use of chitosan gallic acid copolymer |
| CN106893004A (en) * | 2017-03-17 | 2017-06-27 | 广西大学 | A kind of preparation method of shitosan phenolic acid conjugates |
| CN109336081A (en) * | 2018-08-29 | 2019-02-15 | 北京化工大学常州先进材料研究院 | A kind of preparation method of the lithium ion battery spherical carbon cathode based on chitosan |
| CN111514097A (en) * | 2020-05-14 | 2020-08-11 | 燕山大学 | Preparation method of pH-responsive nano hydrogel of walnut shell polyphenol |
| CN111961263A (en) * | 2020-08-28 | 2020-11-20 | 浙江奚态生物科技有限公司 | Preparation method of antibacterial degradable food packaging film material |
Non-Patent Citations (3)
| Title |
|---|
| 杨铭主编: "《纤维增强纳米羟基磷灰石及其生物复合材料》", vol. 01, 中国协和医科大学联合出版社, pages: 72 - 18 * |
| 杨铭主编: "《药物研究中的分子识别》", 中国协和医科大学联合出版社 * |
| 魏子淏: ""乳蛋白-EGCG和壳聚糖-绿原酸复合物的制备、结构表征及功能评价"", 《中国优秀硕士学位论文全文数据库工程科技Ⅰ辑》, no. 8, 15 August 2015 (2015-08-15), pages 024 - 85 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116554704A (en) * | 2023-04-17 | 2023-08-08 | 深圳力越新材料有限公司 | Liquid crystal polymer material for 5G and preparation method thereof |
| CN116554363A (en) * | 2023-04-18 | 2023-08-08 | 珠海市自然之旅生物技术有限公司 | Torone-chitosan derivative and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115124630B (en) | 2024-02-23 |
| WO2022165940A1 (en) | 2022-08-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115124630B (en) | Chitosan derivative and preparation method and application thereof | |
| Sun et al. | Synthesis, characterization, and antimicrobial activities of sulfonated chitosan | |
| Chen et al. | Application of functionalized chitosan in food: A review | |
| Ahmed et al. | Chitosan: a natural antimicrobial agent-a review | |
| EP1483299B1 (en) | Cell wall derivatives from biomass and preparation thereof | |
| Dang et al. | Preparation, characterization, and evaluation of 3, 6-ON-acetylethylenediamine modified chitosan as potential antimicrobial wound dressing material | |
| El-Aidie | A review on chitosan: ecofriendly multiple potential applications in the food industry | |
| Elbarbary et al. | Effect of γ-rays on carboxymethyl chitosan for use as antioxidant and preservative coating for peach fruit | |
| Savin et al. | Antioxidant, cytotoxic and antimicrobial activity of chitosan preparations extracted from Ganoderma lucidum mushroom | |
| Cai et al. | Preparation and characterization of N-benzoyl-O-acetyl-chitosan | |
| EP2451845B1 (en) | Preservatives from chitin derivatives | |
| Nadarajah | Development and characterization of antimicrobial edible films from crawfish chitosan | |
| Liu et al. | Recent advances in chitosan based bioactive materials for food preservation | |
| CN113248632A (en) | Multifunctional traditional Chinese medicine organic acid grafted chitosan quaternary ammonium salt broad-spectrum antibacterial agent and preparation method thereof | |
| Omer et al. | Preparation of Isatin/chitosan schiff base as novel antibacterial biomaterials | |
| CN110558376B (en) | Preparation method and application of chitosan-Zn composite nanoparticle preservative film | |
| WO1991010435A1 (en) | Chitosan pyrithione as antimicrobial agent useful in personal care products | |
| RU2352126C2 (en) | Protective film-forming coating for meat and meat products (versions) | |
| CN115181195B (en) | Quaternary ammonium salt derivative of chitosan oligosaccharide containing quinoline group, preparation method and application | |
| Dutta et al. | 15 Antimicrobial Activity of Chitin, Chitosan, and Their Oligosaccharides | |
| US20140328974A1 (en) | Winemaking method without the admixture of sulphur dioxide, using chitosan-based films | |
| CN110183546B (en) | Gel polysaccharide water-soluble antibacterial derivative and preparation method thereof | |
| CN109354632B (en) | Carboxymethyl aminopolysaccharide derivative and preparation method and application thereof | |
| CN116162181B (en) | Chitosan-tricarboxylic acid derivative and preparation method and application thereof | |
| KR101113729B1 (en) | Process for Preparation of Middle Molecular Chitosan Having Antibacterial Activity against Antibiotic Resistance Bacteria and Uses thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Wu Shigui Inventor after: Mu Bo Inventor after: Zhang Jing Inventor before: Wu Shigui Inventor before: Joshua Johannes van Mullem Inventor before: Zhang Jing |
|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |