CN107467662B - Water-soluble quercetin nanoparticle and preparation method thereof - Google Patents
Water-soluble quercetin nanoparticle and preparation method thereof Download PDFInfo
- Publication number
- CN107467662B CN107467662B CN201710717633.8A CN201710717633A CN107467662B CN 107467662 B CN107467662 B CN 107467662B CN 201710717633 A CN201710717633 A CN 201710717633A CN 107467662 B CN107467662 B CN 107467662B
- Authority
- CN
- China
- Prior art keywords
- quercetin
- water
- wheat protein
- solution
- soluble
- 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.)
- Active
Links
- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 title claims abstract description 220
- 229960001285 quercetin Drugs 0.000 title claims abstract description 118
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 title claims abstract description 110
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 title claims abstract description 110
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 title claims abstract description 110
- 235000005875 quercetin Nutrition 0.000 title claims abstract description 110
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 72
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 241000209140 Triticum Species 0.000 claims abstract description 47
- 235000021307 Triticum Nutrition 0.000 claims abstract description 47
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 47
- 239000000243 solution Substances 0.000 claims abstract description 41
- 239000002245 particle Substances 0.000 claims abstract description 33
- 108010009736 Protein Hydrolysates Proteins 0.000 claims abstract description 26
- 239000003531 protein hydrolysate Substances 0.000 claims abstract description 26
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 20
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 20
- 239000008367 deionised water Substances 0.000 claims abstract description 19
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 19
- 239000012452 mother liquor Substances 0.000 claims abstract description 12
- 108090000790 Enzymes Proteins 0.000 claims abstract description 9
- 102000004190 Enzymes Human genes 0.000 claims abstract description 9
- 238000004108 freeze drying Methods 0.000 claims abstract description 9
- 102000004142 Trypsin Human genes 0.000 claims abstract description 6
- 108090000631 Trypsin Proteins 0.000 claims abstract description 6
- 238000010438 heat treatment Methods 0.000 claims abstract description 6
- 239000012588 trypsin Substances 0.000 claims abstract description 6
- 239000003960 organic solvent Substances 0.000 claims abstract description 5
- 239000010413 mother solution Substances 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 18
- 239000006185 dispersion Substances 0.000 claims description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 15
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 238000000502 dialysis Methods 0.000 claims description 11
- 239000012528 membrane Substances 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 11
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 10
- 238000009826 distribution Methods 0.000 claims description 9
- 239000000693 micelle Substances 0.000 claims description 9
- 238000001471 micro-filtration Methods 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 6
- 229960000583 acetic acid Drugs 0.000 claims description 5
- 238000009835 boiling Methods 0.000 claims description 5
- 239000012362 glacial acetic acid Substances 0.000 claims description 5
- 238000005286 illumination Methods 0.000 claims description 4
- 230000009849 deactivation Effects 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- 239000000126 substance Substances 0.000 abstract description 13
- 230000003993 interaction Effects 0.000 abstract description 7
- 238000011068 loading method Methods 0.000 abstract description 5
- 239000000084 colloidal system Substances 0.000 abstract description 4
- 238000012545 processing Methods 0.000 abstract description 3
- 239000000047 product Substances 0.000 abstract description 3
- 235000012041 food component Nutrition 0.000 abstract description 2
- 230000000415 inactivating effect Effects 0.000 abstract description 2
- 239000005417 food ingredient Substances 0.000 abstract 1
- 239000008187 granular material Substances 0.000 abstract 1
- 230000000052 comparative effect Effects 0.000 description 22
- 239000002131 composite material Substances 0.000 description 16
- 235000018102 proteins Nutrition 0.000 description 14
- 230000000694 effects Effects 0.000 description 10
- 230000007928 solubilization Effects 0.000 description 8
- 238000005063 solubilization Methods 0.000 description 8
- 230000007071 enzymatic hydrolysis Effects 0.000 description 5
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 5
- 239000000413 hydrolysate Substances 0.000 description 5
- 230000002209 hydrophobic effect Effects 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 229940098773 bovine serum albumin Drugs 0.000 description 4
- 229920001184 polypeptide Polymers 0.000 description 4
- 102000004196 processed proteins & peptides Human genes 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 230000017854 proteolysis Effects 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 2
- 108010068370 Glutens Proteins 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 238000002441 X-ray diffraction Methods 0.000 description 2
- 239000012296 anti-solvent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 238000000149 argon plasma sintering Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 229910052802 copper Inorganic materials 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- -1 flavonoid compound Chemical class 0.000 description 2
- 238000012417 linear regression Methods 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- HZWWPUTXBJEENE-UHFFFAOYSA-N 5-amino-2-[[1-[5-amino-2-[[1-[2-amino-3-(4-hydroxyphenyl)propanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoic acid Chemical compound C1CCC(C(=O)NC(CCC(N)=O)C(=O)N2C(CCC2)C(=O)NC(CCC(N)=O)C(O)=O)N1C(=O)C(N)CC1=CC=C(O)C=C1 HZWWPUTXBJEENE-UHFFFAOYSA-N 0.000 description 1
- 206010006458 Bronchitis chronic Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108010061711 Gliadin Proteins 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229930014669 anthocyanidin Natural products 0.000 description 1
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 1
- 235000008758 anthocyanidins Nutrition 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 206010006451 bronchitis Diseases 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 208000007451 chronic bronchitis Diseases 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 230000002888 effect on disease Effects 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000014106 fortified food Nutrition 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 235000021312 gluten Nutrition 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/185—Vegetable proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Botany (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a water-soluble quercetin nanoparticle and a preparation method thereof. Uniformly dispersing wheat protein in deionized water, heating, adjusting the pH value, adding trypsin for enzymolysis, inactivating enzyme, and dialyzing to obtain a wheat protein zymolyte; then, selecting an organic solvent to prepare a quercetin mother solution; dissolving the wheat protein hydrolysate in a PBS solution; dripping the quercetin mother liquor into the wheat protein hydrolysate solution, and centrifuging to obtain a wheat protein hydrolysate-quercetin nanoparticle solution; freeze drying to obtain water soluble quercetin nanometer granule. The invention utilizes the non-covalent interaction between the wheat protein hydrolysate and the quercetin to prepare the water-soluble quercetin nano-particles, the obtained product has good solubility, small particle size, high loading rate, good chemical stability and colloid stability, greatly improves the processing stability of the quercetin, and can be used as food ingredients.
Description
Technical Field
The invention relates to quercetin, in particular to a water-soluble quercetin nanoparticle and a preparation method thereof; the water-soluble quercetin nanoparticles can be used in health food and functional food.
Background
Quercetin (Que) is a flavonoid compound from leguminous plants, has a certain treatment effect on diseases such as chronic bronchitis, coronary heart disease, hypertension and the like, and is a nutritional active ingredient with high utilization value. However, due to poor solubility and chemical stability of Que in water, its application in biological and food fields is limited. Researches show that the nanoparticle delivery system can effectively improve the water solubility and chemical stability of hydrophobic substances and is an effective means for delivering hydrophobic active substances. The current common method is to utilize the non-covalent interaction (including hydrophobic interaction, hydrogen bond, electrostatic interaction, pi-pi accumulation and the like) between the active groups of the protein and the bioactive components to form a colloid conveying system, thereby achieving the purpose of conveying the hydrophobic active nutritional components.
In order to improve the water solubility of quercetin, many researchers have improved the processing characteristics and bioavailability of quercetin by constructing different delivery vehicles. Liujian, etc. { Liujian, Zixiao, Zhouyi, etc. comparison of the interaction patterns of bovine serum albumin with quercetin and anthocyanidin and their nanoparticle characteristics [ J]7-13 } embedding quercetin by Bovine Serum Albumin (BSA) to construct a colloid conveying system, wherein the obtained BSA-Que composite nano-particles have low inoxidizability, and the DPPH free radical clearance rate and ABTS + clearance rate are 53.1% and 85% respectively; mitali Kakran et al { Mitali Kakran, Nanda Gopal Sahoo, Lin Lia, et al]Powder Technology, 2012, 233:59-64 } prepares quee nanoparticles by an anti-solvent precipitation method, with enhanced solubility in water, but with insignificant effect (about 50%); avnesh Kumari et al { Avnesh Kumari, Sudesh Kumar Yadav, Yogish B]The gels and Surfaces B Biointerfaces, 2010, 80(2): 184-;
etc. last page
Emilli，Khalil，et al.Bovine Serum Albumin Nanoparticles Containing Quercetin:Characterization and Antioxidant Activity[J]Journal of Nanoscience and Nanotechnology, 2016, 16: 1346-.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention aims to provide a preparation method of water-soluble quercetin nanoparticles with good solubility, small average particle size, good dispersibility, and good colloidal stability and chemical stability.
Another object of the present invention is to provide water-soluble quercetin nanoparticles prepared by the above method.
The method overcomes the problems of extremely low solubility, poor dispersibility and weak chemical stability of the quercetin in water, and the existing method for preparing the quercetin nanoparticles has the defects of complex preparation process, large particle size of the obtained particles, expensive carrier raw materials and the like.
The purpose of the invention is realized by the following technical scheme:
a preparation method of water-soluble quercetin nanoparticles comprises the following steps:
(1) uniformly dispersing wheat protein in deionized water to prepare a dispersion liquid; heating the material in a water bath to 36-39 ℃, adjusting the pH value to 7-9, adding 0.5-2% of trypsin by mass for enzymolysis, and carrying out enzymolysis for 0.5-3 h under the condition of maintaining the temperature and the pH constant; after the enzymolysis is finished, inactivating the enzyme for 10min in boiling water bath; adjusting the pH value of the material to 6.5-7.5 after enzyme deactivation; centrifuging to remove residues, collecting supernatant, dialyzing with microfiltration membrane, and freeze drying to obtain wheat protein hydrolysate;
(2) selecting a quercetin mother solution prepared from an organic solvent; dissolving the wheat protein hydrolysate in a PBS solution; dropwise adding the quercetin mother liquor into the wheat protein hydrolysate solution under stirring to make the final mass concentration of quercetin 133 mug/mL, and centrifuging to remove the non-embedded quercetin micelles to obtain the wheat protein hydrolysate-quercetin nanoparticle solution; freeze-drying the prepared wheat protein hydrolysate-quercetin nanoparticle solution to obtain water-soluble quercetin nanoparticles.
To further achieve the object of the present invention, preferably, in the step (1), the mass concentration of the dispersion is 3% to 5% (w/v); the pH value is adjusted to be 7-9 by 1-2M NaOH; the pH value of the material is adjusted to 6.5-7.5 by adding 1-2M HCl; the rotational speed of the hydrolysate centrifugation is 10000r/min, and the time is 20 min.
Preferably, in the step (1), the enzymolysis time is 1, 2, 3 h.
Preferably, in the step (1), the dialysis is to filter the supernatant through a microfiltration membrane, then put the filtrate into a dialysis bag with the molecular weight cutoff of 8-14 kDa, dialyze the supernatant for 3d (1 d: deionized water; 2 d: 0.05% glacial acetic acid; 3 d: deionized water) at 4 ℃, and change the dialysate every 8 hours.
Preferably, the membrane pores of the microfiltration membrane have a diameter of 0.45 μm.
Preferably, the organic solvent is ethanol; the concentration of the quercetin mother liquor is 1-4 mg/mL; the concentration of the PBS solution is 5-20 mM, and the pH value is 6.5-8.5. The solubility of quercetin in absolute ethyl alcohol is high, absolute ethyl alcohol is selected for preparing the quercetin mother liquor, and in order to reduce the concentration of ethanol in the quercetin nanoparticle dispersion liquid to the maximum extent, the concentration of the quercetin mother liquor is preferably controlled to be 4mg/mL, and the pH value is 7.0.
Preferably, the mass concentration of the wheat protein hydrolysate dissolved in the PBS solution is 1-5 mg/ml.
Preferably, the step (2) of centrifuging to remove the non-embedded quercetin micelles is carried out at 4 ℃ and 10000r/min for 5 min.
A water soluble quercetin nanoparticle is prepared by the above preparation method; the solubility of the water-soluble quercetin particles in water is 5-16 times higher than that of pure quercetin.
A water soluble quercetin nanoparticle is prepared by the above preparation method; the water-soluble quercetin nanoparticles have good dispersibility, the particle size is below 100nm, and the particle size distribution is uniform.
A water soluble quercetin nanoparticle is prepared by the above preparation method; after the water-soluble quercetin nanoparticles are redissolved, the solution is placed for 8 hours under the conditions of illumination and light shielding, and the residue rate of the quercetin is still over 50 percent, which shows that the water-soluble quercetin nanoparticles have good short-term storage stability.
The raw material used in the present invention may be wheat gluten, commercial gliadins and glutenins.
The invention prepares the wheat protein hydrolysate by enzymolysis, prepares the large protein hydrolysate-quercetin nanoparticle by a simple nanoparticle preparation method of non-covalent interaction of the wheat hydrolysate and quercetin and an anti-solvent, when the concentration of quercetin molecules in water is higher than the critical micelle concentration, the wheat protein hydrolysate can inhibit the formation of micelle, and forms a nano compound with the quercetin by the non-covalent interaction to completely wrap the nano compound, thereby increasing the solubility of the nano compound in water. In addition, because the quercetin is embedded in the protein hydrolysate, oxygen is isolated, and illumination is reduced, so that the chemical stability of the quercetin is improved, and the physiological activity of the quercetin is maintained. And the prepared nano-particles have good colloidal stability and keep nano-size.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the water-soluble quercetin nanoparticles prepared by the method have the characteristics of small particle size, high loading rate and good water solubility.
(2) The water-soluble quercetin nanoparticles prepared by the method have the advantages of average particle size of less than 100nm, uniform particle size distribution, good dispersibility and good colloid stability.
(3) Compared with the quercetin micelle dispersion liquid, the water-soluble quercetin nanoparticle prepared by the invention has higher chemical stability, and the residue rate of quercetin is still kept above 50% after the water-soluble quercetin nanoparticle is placed for 8 hours under the conditions of illumination and light shielding.
(4) The invention provides a method for preparing quercetin nanoparticles, which greatly improves the water solubility of quercetin, obviously improves the processing performance of the quercetin, and realizes the industrial production of quercetin-enriched food. And greatly improves the added value of the corn protein.
(5) The invention relates to only two raw materials: the quercetin and the wheat gluten protein have low cost of raw materials and wide sources; and the preparation process of the quercetin nanoparticles is simple and easy to control, economical, practical, green and safe, and is suitable for industrial continuous production.
Drawings
FIG. 1 is a solution observation diagram of solubilization of Que in comparative example 1 and examples 1 to 3;
FIG. 2 is a graph showing the effect of WPH concentration on solubilization of Que in comparative example 1 and examples 4-7;
FIG. 3A is a graph showing the distribution of the particle size of quercetin nanoparticles in comparative example 1 and examples 4 to 7;
FIG. 3B is a graph showing the analysis of the results of the average particle size of quercetin nanoparticles in comparative example 1 and examples 4 to 7;
fig. 4 is a graph of analysis of the results of short term storage chemical stability of quercetin particles in comparative example 1 and example 7;
FIG. 5 is an X-ray diffraction pattern of Que composite nanoparticles in comparative example 1(Que), comparative example 2(WPH), and example 7.
Detailed Description
The invention will be further described with reference to the following examples for better understanding, but the scope of the invention as claimed is not limited to the examples.
Comparative example 1
Firstly, preparing 4mg/mL quercetin mother liquor by using absolute ethyl alcohol as a solvent. The quercetin stock solution was added dropwise to a 10mM, pH7.0 PBS solution under rapid stirring to give a final concentration of quercetin of 133. mu./mL. Then the mixture is stirred for 30min at normal temperature, and the prepared nano-particle solution is stored at 4 ℃ in the dark.
Comparative example 2
Firstly, uniformly dispersing wheat protein in deionized water to prepare 5% (w/v, g/mL) of dispersion liquid. Heating the dispersion in water bath to 37 deg.C, adjusting pH to 7.0 with 1M NaOH, adding 2% trypsin for enzymolysis, and performing enzymolysis for 3 hr under the condition of maintaining constant temperature and pH. After completion of the enzymatic hydrolysis, the batch was adjusted to pH 6.5 with 1M HCl and then heated in a boiling water bath for 10min to inactivate the enzyme. Centrifuging the hydrolysate at 10000r/min for 20min, filtering the supernatant with 0.45 μm microfiltration membrane, loading into dialysis bag with molecular weight cutoff of 14kDa, dialyzing in deionized water at 4 deg.C for 3d (1 d: deionized water; 2 d: 0.05% glacial acetic acid; 3 d: deionized water), and replacing dialysate every 8 h. After dialysis, carrying out freeze drying treatment to obtain a wheat protein hydrolysate; storing for later use.
Next, the wheat protein hydrolysate was dissolved in a 20mM PBS solution at a concentration of 5 mg/ml.
Example 1
A preparation method of water-soluble quercetin nanoparticles comprises the following steps:
the method comprises the following steps: uniformly dispersing wheat protein in deionized water to prepare 5% (w/v, g/mL) of dispersion liquid. Heating the dispersion in water bath to 37 deg.C, adjusting pH to 9.0 with 1M NaOH, adding 1% trypsin by mass, and performing enzymolysis for 1 hr while maintaining constant temperature and pH. After completion of the enzymatic hydrolysis, the batch was adjusted to pH 7.5 with 1M HCl and then heated in a boiling water bath for 10min to inactivate the enzyme. Centrifuging the hydrolysate at 10000r/min for 20min, filtering the supernatant with 0.45 μm microfiltration membrane, loading into dialysis bag with molecular weight cutoff of 14kDa, dialyzing in deionized water at 4 deg.C for 3d (1 d: deionized water; 2 d: 0.05% glacial acetic acid; 3 d: deionized water), and replacing dialysate every 8 h. After dialysis, carrying out freeze drying treatment to obtain a wheat protein hydrolysate; storing for later use.
Step two: firstly, preparing 4mg/mL quercetin mother liquor by using absolute ethyl alcohol as a solvent. Wheat protein hydrolysate was dissolved in 10mM PBS at a concentration of 5 mg/ml. Dropping the quercetin mother liquor into the wheat protein hydrolysate solution under rapid stirring to make the final mass concentration of quercetin be 100 μ g/mL. Stirring at room temperature for 30min, centrifuging at 4 deg.C and 10000r/min for 5min, removing the non-embedded quercetin micelle, and storing the prepared wheat protein hydrolysate-quercetin nanoparticle solution at 4 deg.C in dark place.
Example 2:
a method for preparing water-soluble quercetin nanoparticles, wherein enzymolysis time is controlled to be 2h, and the other steps are the same as those in example 1.
Example 3:
a method for preparing water-soluble quercetin nanoparticles, wherein enzymolysis time is controlled to be 3h, and the other steps are the same as those in example 1.
Fig. 1 is a solution observation diagram of solubilization conditions of PBS solution and WPH subjected to enzymolysis for 1h, 2h and 3h on Que, namely, quercetin nanoparticle dispersion liquids prepared in comparative example 1 and examples 1 to 3. As can be seen from FIG. 1, the solubilization of quercetin by the enzymatic hydrolysis products varies with the enzymatic hydrolysis time. When the solution is PBS solution (enzymolysis time is 0h), the quercetin is turbid in the water solution, and floccules float in the solution. Along with the extension of the enzymolysis time to 1-3 h, the quercetin and enzymolysis product composite solution is gradually clarified, and when the enzymolysis is carried out for 2h, the solution is basically free of precipitation, because the restriction enzyme hydrolysis of wheat protein is possibly favorable for obtaining the polypeptide with better amphipathy, and the polypeptide has a good embedding effect on hydrophobic polyphenol.
Example 4
A preparation method of water-soluble quercetin nanoparticles comprises the following steps:
the method comprises the following steps: uniformly dispersing wheat protein in deionized water to prepare 3% (w/v, g/mL) of dispersion liquid. Heating the dispersion in water bath to 37 deg.C, adjusting pH to 8.0 with 1M NaOH, adding 0.5% trypsin for enzymolysis, and performing enzymolysis for 3 hr under the condition of maintaining constant temperature and pH. After completion of the enzymatic hydrolysis, the batch was adjusted to pH7 with 1M HCl and then heated in a boiling water bath for 10min to inactivate the enzyme. Centrifuging the hydrolysate at 10000r/min for 20min, filtering the supernatant with 0.45 μm microfiltration membrane, loading into dialysis bag with molecular weight cutoff of 8kDa, dialyzing in deionized water at 4 deg.C for 3d (1 d: deionized water; 2 d: 0.05% glacial acetic acid; 3 d: deionized water), and replacing dialysate every 8 h. After dialysis, carrying out freeze drying treatment to obtain a wheat protein hydrolysate; storing for later use.
Step two: firstly, preparing 4mg/mL quercetin mother liquor by using absolute ethyl alcohol as a solvent. Wheat protein hydrolysate was dissolved in 20mM PBS at a concentration of 1 mg/ml. Dropping the quercetin mother liquor into the wheat protein hydrolysate solution under rapid stirring to make the final mass concentration of quercetin 133 μ g/mL. Stirring at room temperature for 30min, centrifuging at 8000r/min at 4 deg.C for 15min, removing the non-embedded quercetin micelle, and storing the prepared wheat protein hydrolysate-quercetin nanoparticle solution at 4 deg.C in dark place.
Example 5
A method for preparing water-soluble quercetin nanoparticles comprises dissolving wheat protein hydrolysate in 20mM PBS solution at a concentration of 2mg/ml, and the same as example 4.
Example 6
A method for preparing water-soluble quercetin nanoparticles comprises dissolving wheat protein hydrolysate in 20mM PBS solution at a concentration of 3mg/ml, and the same as example 4.
Example 7
A method for preparing water-soluble quercetin nanoparticles comprises dissolving wheat protein hydrolysate in 20mM PBS solution at a concentration of 5mg/ml, and the same as example 4.
Testing indexes are as follows:
(1) solubility:
standard curve: 0.0208g of quercetin is precisely weighed, absolute ethyl alcohol is used for metering volume to a 100mL volumetric flask to obtain a Que standard solution with the concentration of 208 mu g/mL, the absolute ethyl alcohol is used for preparing different concentrations (0, 2, 4, 6, 8 and 10 mu g/mL), the absolute ethyl alcohol is used for zeroing, an ultraviolet spectrophotometer is used for color comparison at the wavelength of 374nm, the optical density is read, and a linear regression equation is made according to data.
And diluting the Que composite nano-particle samples in the comparative example 1 and the examples 4-7, carrying out color comparison under the same condition, calculating the solubility by using a linear regression equation, and comparing to obtain the solubilization effect of the protein zymolyte on the Que.
(2) Particle size distribution and average particle size:
particle size testing of quercetin composite nanoparticles in comparative example 1 and examples 4-7 was performed at 25 ℃ room temperature, and the effect of large particles was reduced using a back-scattering technique (light scattering angle 173 °). The refractive index of the Que nanoparticles was set to 1.763, the refractive index of a dispersant phosphate buffer (20mM, pH7.2) was 1.33, and the average particle diameter (Z-average), polydispersity index (PDI) and volume distribution of the particles were calculated using the change in light scattering intensity with time and measured in parallel three times.
The results of measuring the solubility, average particle diameter, and dispersion index of the Que composite nanoparticles in comparative example 1 and examples 4 to 7 are shown in table 1.
TABLE 1
As can be seen from Table 1, the solubility of the Que composite nanoparticles in examples 4 to 7 was significantly improved, the average particle size was reduced to about 100nm, the dispersion indexes PDI were all less than 0.5, and good monodispersity was exhibited, as compared to comparative example 1.
FIG. 2 shows the effect of WPH concentrations of 0, 1, 2, 3, and 5mg/ml on the solubilization of Que, i.e., the solubilization of the quercetin dispersions prepared in comparative example 1 and examples 4-7. As can be seen from FIG. 2, with the increasing concentration of the proteolytic enzyme, the composite solution is gradually clear and transparent, and no obvious floccule is seen. Within the scope defined by the claims, the presence of wheat proteolysis will increase the solubility of Que in water significantly (p <0.05) by nearly 5-16 fold. This may be associated with some amphiphilicity of the wheat protein hydrolysate. The quercetin and the amphiphilic polypeptide in the zymolyte may have non-covalent interaction, and self-assembly forms composite nano particles, so that a good solubilization effect is shown for Que.
Fig. 3A is a particle size distribution diagram of the quercetin dispersions prepared in comparative example 1 and examples 4 to 7. Quee in aqueous solution shows a broad particle size distribution, ranging from tens of microns to hundreds of thousands of microns, indicating that colloidal particles of free quee are extremely unstable in aqueous solution; the PDI value of the WHP-Que composite nano-particles is less than 0.5, the particle size distribution is uniform, a single peak is only in the range of 10-100nm, and good monodispersity is shown.
FIG. 3B is a graph showing the results of WPH concentrations of 0, 1, 2, 3, and 5mg/ml on the average particle size of quercetin nanoparticles (i.e., comparative example 1 and examples 4 to 7). As shown in FIG. 3B, the mean particle size of the free quercetin solution reached 1800nm, which is visible to the naked eye; the average particle size of the WHP-Que composite nanoparticles was less than 100nm, which is consistent with the solubility results. The solubility of Que was improved by Avnesh et al using poly D, l-lactide (PLA) and by Denitsa et al using chitosan-alginate, but the formed nanoparticles all had larger particle sizes (around 130 + -30 nm and 900nm, respectively).
Fig. 4 is a graph showing the analysis of the chemical stability of water-soluble quercetin particles during short-term storage, which is obtained by measuring the content of (0, 1, 2, 3, 4, 6, 8h) quee (determination of solubility as a measure of quee content) by allowing the WPH-quee composite nanoparticle solution prepared in example 7 and a quee aqueous solution (comparative example 1) to stand under both light and dark environments. As can be seen from FIG. 4, the storage stability of Que was also improved to some extent in the presence of wheat proteolytic enzyme. In the WPH solution, the residue rate of Que is more than 50% after the Que is placed for 8 hours. The reason why the proteolysis substance provides protection for the chemical stability of the Que may be that the polypeptide units in the wheat proteolysis substance are combined with the Que to generate a 'solidification effect' which can reduce the activity of the Que and thus reduce the reaction activity of the Que, or the Que is embedded to block the contact of the Que and an oxidant and the like in a physical mode. It should be noted that the antioxidant activity of the wheat protein zymolyte itself is also likely to participate in the oxidation protection of Que.
Fig. 5 is an X-ray diffraction pattern of the quee (comparative example 1), WPH (comparative example 2) and the quee composite nanoparticles of example 7, which was obtained by freeze-drying the samples prepared in comparative example 1, comparative example 2 and example 7 under normal temperature conditions and performing XRD measurement. (test indexes are as follows: use of D8ADVANCE diffractor (Bruker, Germany), copper target K copper radiation, target using kerANCE, LynxExe array detector, slit DS 1 mm; RS 8mm, Ni filter, tube pressure 40KV, tube flow 40mA, scanning wave). As can be seen from FIG. 5, Que has a large number of crystal peaks on the graph, and no obvious crystal peak appears in WPH and composite nanoparticles, which indicates that free Que exists in water in a highly crystalline state, and is converted into an amorphous state after WPH forms composite nanoparticles, indicating that Que and WPH form a stable non-covalent composite with an amorphous structure.
Claims (7)
1. A preparation method of water-soluble quercetin nanoparticles is characterized by comprising the following steps:
(1) uniformly dispersing wheat protein in deionized water to prepare a dispersion liquid; heating the material in a water bath to 36-39 ℃, adjusting the pH value to 7-9, adding 0.5-2% of trypsin by mass for enzymolysis, and carrying out enzymolysis for 1-3 h under the condition of maintaining the temperature and the pH constant; after the enzymolysis is finished, enzyme is deactivated; adjusting the pH value of the material to 6.5-7.5; centrifuging to remove residues, collecting supernatant, dialyzing with microfiltration membrane, and freeze drying to obtain wheat protein hydrolysate; and (3) the dialysis is to filter the supernatant through a microfiltration membrane, then put the supernatant into a dialysis bag with the molecular weight cutoff of 8-14 kDa, and dialyze the supernatant for 3d at 4 ℃, wherein the content of the supernatant in the dialysis bag is as follows: deionized water; and 2 d: 0.05% glacial acetic acid; and 3 d: deionized water, and the dialysate is replaced every 8 hours; the diameter of the membrane pores of the microfiltration membrane is 0.45 mu m;
(2) selecting an organic solvent to prepare a quercetin mother solution; dissolving the wheat protein hydrolysate in a PBS solution; dropwise adding the quercetin mother liquor into the wheat protein hydrolysate solution under stirring to enable the final mass concentration of quercetin to be 100-133 mug/mL, and centrifuging to remove the non-embedded quercetin micelles to obtain a wheat protein hydrolysate-quercetin nanoparticle solution; freeze-drying the prepared wheat protein hydrolysate-quercetin nanoparticle solution to obtain water-soluble quercetin nanoparticles; the organic solvent is absolute ethyl alcohol; the concentration of the quercetin mother liquor is 1-4 mg/mL; the concentration of the PBS solution is 5-20 mM, and the pH value is 6.5-8.5.
2. The preparation method according to claim 1, wherein in the step (1), the mass-to-volume ratio of the dispersion is 3% to 5%, wherein the mass unit is gram and the volume unit is milliliter; the enzyme deactivation is carried out by boiling water bath; the enzyme deactivation time is 5-10 min.
3. The preparation method according to claim 1, wherein in the step (1), the pH value is adjusted to 7-9 by adding 1-2M NaOH; the pH value of the material is adjusted to 6.5-7.5 by adding 1-2M HCl; the rotation speed of the centrifugation is 10000r/min, and the time is 20 min.
4. The method according to claim 1, wherein the mass concentration of the wheat protein hydrolysate dissolved in the PBS solution is 1-5 mg/ml.
5. The method according to claim 1, wherein the step (2) of centrifuging to remove the non-embedded quercetin micelles is carried out at a temperature of 4-8 ℃ and a rotation speed of 8000-10000 r/min for 5-15 min.
6. A water-soluble quercetin nanoparticle solution, which is prepared by the preparation method according to any one of claims 1 to 5; the water-soluble quercetin nanoparticles are water-soluble, and the solubility of the quercetin particles in water is improved by 5-16 times compared with that of pure quercetin.
7. A water-soluble quercetin nanoparticle prepared by the preparation method according to any one of claims 1 to 5; the quercetin nanoparticles have good dispersibility, the particle size is below 100nm, and the particle size distribution is uniform; after the water-soluble quercetin nanoparticles are redissolved, the solution is placed for 8 hours under the conditions of illumination and light shielding, and the residual rate of the quercetin is more than 50%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710717633.8A CN107467662B (en) | 2017-08-21 | 2017-08-21 | Water-soluble quercetin nanoparticle and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710717633.8A CN107467662B (en) | 2017-08-21 | 2017-08-21 | Water-soluble quercetin nanoparticle and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107467662A CN107467662A (en) | 2017-12-15 |
| CN107467662B true CN107467662B (en) | 2021-01-19 |
Family
ID=60600989
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710717633.8A Active CN107467662B (en) | 2017-08-21 | 2017-08-21 | Water-soluble quercetin nanoparticle and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107467662B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2679044C1 (en) * | 2018-03-02 | 2019-02-05 | Александр Александрович Кролевец | Method of obtaining marmelade with nanostructured querticine and dihydroquerticine |
| CN108851084B (en) * | 2018-06-06 | 2021-06-15 | 福建省农业科学院农业工程技术研究所 | Colon positioning micelle loaded with quercetin and preparation method thereof |
| CN111150063A (en) * | 2020-01-07 | 2020-05-15 | 天津科技大学 | Method for simultaneously improving hydrophobicity and stability of water-soluble active ingredients by using cage-shaped plant ferritin and application |
| CN113230234B (en) * | 2021-04-27 | 2022-04-26 | 江苏大学 | Ultrasonic preparation method of bioactive ingredient-loaded protein peptide-polysaccharide nanoparticles |
| CN113575951B (en) * | 2021-08-06 | 2023-06-16 | 合肥工业大学 | Starch-based dual-load functional nanoparticle, and preparation method and application thereof |
| CN114098076B (en) * | 2021-11-18 | 2022-11-01 | 江南大学 | Preparation method of genipin-crosslinked quercetin-zein/pectin/chitosan nanoparticles |
| CN114983974A (en) * | 2022-06-06 | 2022-09-02 | 盐城工业职业技术学院 | Preparation method of wheat alcohol soluble protein-Arabic gum-quercetin nanoparticles |
| CN115124608A (en) * | 2022-06-27 | 2022-09-30 | 上海理工大学 | Barley-derived self-assembly peptide and preparation method and application thereof |
| CN115226895A (en) * | 2022-06-28 | 2022-10-25 | 海南大学 | Preparation method of areca seed nanocellulose-quercetin compound |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105054073A (en) * | 2015-08-10 | 2015-11-18 | 华南理工大学 | Water-soluble vitamin D3 nanometer particles and preparation method thereof |
-
2017
- 2017-08-21 CN CN201710717633.8A patent/CN107467662B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105054073A (en) * | 2015-08-10 | 2015-11-18 | 华南理工大学 | Water-soluble vitamin D3 nanometer particles and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| Evaluation of gliadins nanoparticles as drug delivery systems:a study of three different drugs;C.Duclairoir,et al;《International Journal of Pharmaceutics》;20031231(第253期);第133-144页 * |
| Plant protein-based hydrophobic fine and ultrafine carrier particles in drug delivery systems;Hedieh Malekzad,et al.3;《Critical Reviews in Biotechnology》;20170424;第38卷(第1期);第47-67页，尤其是摘要，第9页最后1段，第10页第1、4段 * |
| Quercetin loaded biopolymeric colloidal particles prepared by simultaneous precipitation of quercetin with hydrophobic protein inaqueous medium;Patel A R et al.;《Food Chemistry》;20121231;第133卷(第2期);第423-429页，尤其是摘要，第424页第2.2节 * |
| 酶法复合改性对小麦面筋蛋白性质和结构的影响;王凯强等;《现代食品科技》;20161231;第32卷(第3期);第177-182，280页，尤其是摘要，第178页第1.2.1节 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107467662A (en) | 2017-12-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107467662B (en) | Water-soluble quercetin nanoparticle and preparation method thereof | |
| Thandapani et al. | Size optimization and in vitro biocompatibility studies of chitosan nanoparticles | |
| CN112022834B (en) | Hyperoside-loaded zein-pectin composite nano-particles and preparation method thereof | |
| Li et al. | The characterization and stability of the soy protein isolate/1-Octacosanol nanocomplex | |
| Sun et al. | Physical, structural, thermal and morphological characteristics of zeinquercetagetin composite colloidal nanoparticles | |
| Zhang et al. | Preparation and characterization of selenium nanoparticles decorated by Spirulina platensis polysaccharide | |
| CN110623937A (en) | Tea polyphenol-based multifunctional nano-composite as well as preparation method and application thereof | |
| CN105054073B (en) | A kind of water miscible vitamine D3 nano particle and preparation method thereof | |
| Bhatia et al. | Formulation and optimization of quinoa starch nanoparticles: Quality by design approach for solubility enhancement of piroxicam | |
| CN112205628A (en) | Composite condensate with double embedding functions and preparation method and application thereof | |
| Zhang et al. | The preparation of chitosan nanoparticles by wet media milling | |
| Wani et al. | Techno-functional characterization of chitosan nanoparticles prepared through planetary ball milling | |
| Cai et al. | Effect of molecular weight of chitosan on the formation and properties of zein-nisin-chitosan nanocomplexes | |
| An et al. | Toward function starch nanogels by self-assembly of polysaccharide and protein: From synthesis to potential for polyphenol delivery | |
| Niu et al. | Hydrophobin-enhanced stability, dispersions and release of curcumin nanoparticles in water | |
| Cheng et al. | pH-induced complex coacervation of fish gelatin and carboxylated chitosan: Phase behavior and structural properties | |
| Kou et al. | Preparation and application of a polymer with pH/temperature-responsive targeting | |
| CN113368076B (en) | Protein microparticles and methods of making and using the same | |
| CN112121178B (en) | Water-soluble zein-EGCG covalent compound and preparation and application thereof | |
| CN113633781A (en) | Astaxanthin-chitosan self-assembly nano compound and preparation method and application thereof | |
| Kumar et al. | Formulation and evaluation of quercetin-loaded banana starch nanoparticles | |
| Han et al. | Synthesis and evaluation of hydroxycamptothecin-encapsulated chitosan nanospheres for the treatment of liver cancer | |
| John et al. | Cross-linking with multifunctional excipients and its effect on the physicochemical properties and release profile of ibuprofen-loaded Digitaria exilis starch nanoparticles | |
| Bo et al. | Immunoregulatory effects on RAW264. 7 cells and subacute oral toxicity of ultra-large pore mesoporous silica nanoparticles loading Lycium barbarum polysaccharides | |
| CN114504565B (en) | Tumor microenvironment responsive intelligent chitosan arsenic-loaded nanoparticle and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 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 |