CN114698688B - Preparation and application of nanostructure lipid carrier - Google Patents
Preparation and application of nanostructure lipid carrier Download PDFInfo
- Publication number
- CN114698688B CN114698688B CN202210450397.9A CN202210450397A CN114698688B CN 114698688 B CN114698688 B CN 114698688B CN 202210450397 A CN202210450397 A CN 202210450397A CN 114698688 B CN114698688 B CN 114698688B
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- CN
- China
- Prior art keywords
- medium
- oil
- essential oil
- lipid carrier
- litsea cubeba
- Prior art date
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- 150000002632 lipids Chemical class 0.000 title claims abstract description 66
- 239000002086 nanomaterial Substances 0.000 title claims abstract description 17
- 238000002360 preparation method Methods 0.000 title abstract description 24
- 239000000341 volatile oil Substances 0.000 claims abstract description 111
- 240000002262 Litsea cubeba Species 0.000 claims abstract description 79
- 235000012854 Litsea cubeba Nutrition 0.000 claims abstract description 79
- 239000003921 oil Substances 0.000 claims abstract description 47
- 235000019198 oils Nutrition 0.000 claims abstract description 47
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 43
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- 238000001816 cooling Methods 0.000 claims abstract description 6
- 238000000265 homogenisation Methods 0.000 claims abstract description 3
- 239000012071 phase Substances 0.000 claims description 66
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- 238000000034 method Methods 0.000 claims description 27
- 239000004006 olive oil Substances 0.000 claims description 22
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- 102000004882 Lipase Human genes 0.000 claims description 21
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- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 claims description 19
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- 239000000123 paper Substances 0.000 claims description 17
- 150000004668 long chain fatty acids Chemical class 0.000 claims description 16
- 150000004667 medium chain fatty acids Chemical class 0.000 claims description 16
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 16
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- 238000006555 catalytic reaction Methods 0.000 claims description 11
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- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 claims description 10
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- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 claims description 9
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- 239000007908 nanoemulsion Substances 0.000 description 14
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
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- 238000002835 absorbance Methods 0.000 description 4
- 239000000969 carrier Substances 0.000 description 4
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- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 4
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- 230000004580 weight loss Effects 0.000 description 4
- 244000178231 Rosmarinus officinalis Species 0.000 description 3
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- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 3
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- WTEVQBCEXWBHNA-UHFFFAOYSA-N Citral Natural products CC(C)=CCCC(C)=CC=O WTEVQBCEXWBHNA-UHFFFAOYSA-N 0.000 description 2
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- 235000001636 Mentha x rotundifolia Nutrition 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 230000002292 Radical scavenging effect Effects 0.000 description 2
- QNVSXXGDAPORNA-UHFFFAOYSA-N Resveratrol Natural products OC1=CC=CC(C=CC=2C=C(O)C(O)=CC=2)=C1 QNVSXXGDAPORNA-UHFFFAOYSA-N 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- LUKBXSAWLPMMSZ-OWOJBTEDSA-N Trans-resveratrol Chemical compound C1=CC(O)=CC=C1\C=C\C1=CC(O)=CC(O)=C1 LUKBXSAWLPMMSZ-OWOJBTEDSA-N 0.000 description 2
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- 239000012043 crude product Substances 0.000 description 2
- GHVNFZFCNZKVNT-UHFFFAOYSA-N decanoic acid Chemical compound CCCCCCCCCC(O)=O GHVNFZFCNZKVNT-UHFFFAOYSA-N 0.000 description 2
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- WTEVQBCEXWBHNA-JXMROGBWSA-N geranial Chemical compound CC(C)=CCC\C(C)=C\C=O WTEVQBCEXWBHNA-JXMROGBWSA-N 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- XMGQYMWWDOXHJM-UHFFFAOYSA-N limonene Chemical compound CC(=C)C1CCC(C)=CC1 XMGQYMWWDOXHJM-UHFFFAOYSA-N 0.000 description 2
- CDOSHBSSFJOMGT-UHFFFAOYSA-N linalool Chemical compound CC(C)=CCCC(C)(O)C=C CDOSHBSSFJOMGT-UHFFFAOYSA-N 0.000 description 2
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- 235000021283 resveratrol Nutrition 0.000 description 2
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- 238000000108 ultra-filtration Methods 0.000 description 2
- 239000001490 (3R)-3,7-dimethylocta-1,6-dien-3-ol Substances 0.000 description 1
- CDOSHBSSFJOMGT-JTQLQIEISA-N (R)-linalool Natural products CC(C)=CCC[C@@](C)(O)C=C CDOSHBSSFJOMGT-JTQLQIEISA-N 0.000 description 1
- 241000228245 Aspergillus niger Species 0.000 description 1
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- 239000005632 Capric acid (CAS 334-48-5) Substances 0.000 description 1
- 102000030523 Catechol oxidase Human genes 0.000 description 1
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- 244000175448 Citrus madurensis Species 0.000 description 1
- 201000007336 Cryptococcosis Diseases 0.000 description 1
- 241000221204 Cryptococcus neoformans Species 0.000 description 1
- 241000238557 Decapoda Species 0.000 description 1
- 244000061508 Eriobotrya japonica Species 0.000 description 1
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- 235000017317 Fortunella Nutrition 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 description 1
- 244000246386 Mentha pulegium Species 0.000 description 1
- 235000016257 Mentha pulegium Nutrition 0.000 description 1
- 235000004357 Mentha x piperita Nutrition 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- 235000019483 Peanut oil Nutrition 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 108700023158 Phenylalanine ammonia-lyases Proteins 0.000 description 1
- IMQLKJBTEOYOSI-UHFFFAOYSA-N Phytic acid Natural products OP(O)(=O)OC1C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C1OP(O)(O)=O IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 description 1
- 229920001214 Polysorbate 60 Polymers 0.000 description 1
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- 238000010586 diagram Methods 0.000 description 1
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
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- 235000001050 hortel pimenta Nutrition 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 229940087305 limonene Drugs 0.000 description 1
- 235000001510 limonene Nutrition 0.000 description 1
- 229930007744 linalool Natural products 0.000 description 1
- 239000000944 linseed oil Substances 0.000 description 1
- 235000021388 linseed oil Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000004530 micro-emulsion Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 239000005022 packaging material Substances 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000000312 peanut oil Substances 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 229940085991 phosphate ion Drugs 0.000 description 1
- 235000002949 phytic acid Nutrition 0.000 description 1
- 239000000467 phytic acid Substances 0.000 description 1
- 229940068041 phytic acid Drugs 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000008165 rice bran oil Substances 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
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- 238000002791 soaking Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 235000020238 sunflower seed Nutrition 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- TUNFSRHWOTWDNC-HKGQFRNVSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCC[14C](O)=O TUNFSRHWOTWDNC-HKGQFRNVSA-N 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/154—Organic compounds; Microorganisms; Enzymes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65D—CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
- B65D19/00—Pallets or like platforms, with or without side walls, for supporting loads to be lifted or lowered
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65D—CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
- B65D81/00—Containers, packaging elements, or packages, for contents presenting particular transport or storage problems, or adapted to be used for non-packaging purposes after removal of contents
- B65D81/24—Adaptations for preventing deterioration or decay of contents; Applications to the container or packaging material of food preservatives, fungicides, pesticides or animal repellants
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65D—CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
- B65D85/00—Containers, packaging elements or packages, specially adapted for particular articles or materials
- B65D85/30—Containers, packaging elements or packages, specially adapted for particular articles or materials for articles particularly sensitive to damage by shock or pressure
- B65D85/34—Containers, packaging elements or packages, specially adapted for particular articles or materials for articles particularly sensitive to damage by shock or pressure for fruit, e.g. apples, oranges or tomatoes
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Engineering & Computer Science (AREA)
- Mechanical Engineering (AREA)
- Life Sciences & Earth Sciences (AREA)
- Food Science & Technology (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Fats And Perfumes (AREA)
- Medicinal Preparation (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the field of foods, and discloses a preparation method of a nano-structure lipid carrier based on medium-long chain diglyceride embedded litsea cubeba essential oil and application of the nano-structure lipid carrier in fruit and vegetable fresh-keeping. The preparation method comprises the following steps: taking litsea cubeba essential oil, vegetable oil and medium-long chain diglyceride in a molten state as oil phases; and then mixing the water phase into the oil phase, carrying out high-speed shearing and homogenization, carrying out ultrasonic treatment, stirring in an ice bath, cooling, and obtaining the nano-structure lipid carrier based on the medium-long chain diglyceride embedded litsea cubeba essential oil. The nano-structured lipid carrier for embedding the litsea cubeba essential oil has the advantages of small particle size, high encapsulation efficiency and good stability, solves the difficult problem that the litsea cubeba essential oil is difficult to dissolve in water and has high volatility as oil, realizes efficient embedding and delayed release of the litsea cubeba essential oil, fully exerts broad-spectrum antibacterial property and oxidation resistance, can effectively inhibit the spoilage of blueberries, prolongs the shelf life of the blueberries and improves the economic value.
Description
Technical Field
The invention belongs to the field of foods, and particularly relates to a preparation method and application of a nano-structure lipid carrier based on medium-long chain diglyceride embedded litsea cubeba essential oil.
Background
The litsea cubeba essential oil (Litsea cubeba essential oil) is a yellowish natural aromatic compound extracted from fresh litsea cubeba fruits, has lipophilicity and high volatility, is rich in bioactive components such as citral, limonene, linalool and the like, and has remarkable biological activities such as antifungal, antibacterial, insect-resistant, antioxidant and the like. The litsea cubeba essential oil can destroy cell membranes of bacteria to cause protein denaturation, has obvious inhibition effect on escherichia coli, staphylococcus aureus, cryptococcus neoformans, aspergillus niger, rhizopus oryzae, bacillus subtilis, candida albicans and the like, and has broad-spectrum antibacterial and antifungal effects. The main component citral of the litsea cubeba essential oil can effectively remove DPPH free radicals, hydroxyl free radicals and superoxide free radicals, and has good oxidation resistance. At present, the litsea cubeba essential oil has been partially studied in the field of food preservation and fresh-keeping, and can remarkably inhibit the activities of loquat and kumquat peroxidase, polyphenol oxidase and phenylalanine ammonia lyase and delay the senescence of fruits; the compound preservative prepared from VE and phytic acid can prolong the shelf lives of shrimp meat and pork.
Although the external activity of the litsea cubeba essential oil is obvious and has been partially studied in the field of food preservation, the antioxidant activity of the litsea cubeba essential oil is quickly reduced due to the instability of light, heat and oxygen during the storage process, and the defects of easy volatilization, low water solubility, strong special flavor of the litsea cubeba essential oil and the like are overcome, so that the value of the litsea cubeba essential oil cannot be fully exerted in practical application.
Chinese patent publication (CN 108464998A) discloses a peppermint essential oil emulsion with high stability. The mint essential oil and the resveratrol embedded by the method show synergistic antibacterial effect, but the method uses absolute ethyl alcohol which is an organic solvent, the preparation operation by an anti-solvent method is complex, and the embedding rate of the mint essential oil and the resveratrol is low and is only about 80 percent.
Chinese patent publication (CN 113440479A) discloses a nanoemulsion of plant essential oil, and a preparation method and application thereof. The nanoemulsion prepared by the method has high stability and low volatility, and has obvious inhibition effect on escherichia coli and staphylococcus aureus. However, the emulsion prepared by the method has larger particle size.
Chinese patent publication (CN 111110720A) discloses a rosemary essential oil nano liposome and a preparation method and application thereof. The method overcomes the defects of rosemary essential oil in the using process, has good slow release effect, can increase the retention amount and retention time of the medicament in skin, better plays the local effect of the medicament, and has good application prospect. However, the emulsion prepared by the method has low encapsulation rate on rosemary essential oil.
Disclosure of Invention
In order to overcome the defects of the prior art, the primary aim of the invention is to provide a preparation method of a nano-structured lipid carrier based on the medium-long chain diglyceride embedded litsea cubeba essential oil, which is efficient, simple in process, green, safe, high in stability and high in encapsulation efficiency. The nanostructure lipid carrier provided by the invention comprises an oil phase and a water phase, wherein the oil phase consists of solid lipid (medium-long chain diglyceride) and liquid lipid (vegetable oil and litsea cubeba essential oil), and the water phase comprises a surfactant and a phosphate buffer solution. The litsea cubeba essential oil serving as part of liquid lipid in the system has the dual functions of antioxidation and bacteriostasis and functional liquid oil, can be better wrapped in fat crystal lattices, can reduce lattice order of vegetable oil, reduce the problems of essential oil exudation and the like caused by crystal form conversion, improves the solubility and stability of a nano lipid carrier in water, and can well realize the slow release of the essential oil.
The invention also aims to provide the nano-structured lipid carrier emulsion based on the medium-long chain diglyceride embedded litsea cubeba essential oil, which is prepared by the method, has small and uniform emulsion particle size and high stability in the storage process, and shows the high encapsulation efficiency and the slow release effect which are not possessed by the traditional nano-emulsion and the solid lipid nanoparticle.
The invention also aims to provide the application of the emulsion based on the nano-structured lipid carrier of the medium-long chain diglyceride embedded litsea cubeba essential oil in fruit and vegetable fresh-keeping, in particular to the application in blueberry fresh-keeping. The controllable release performance of the nano-structured lipid carrier as an encapsulating material can fully exert the anti-corrosion potential of litsea cubeba essential oil in fruit and vegetable fresh-keeping, prolong the shelf life of fruits and vegetables and improve the economic value.
The aim of the invention is achieved by the following technical scheme:
the preparation method of the nano-structured lipid carrier based on the embedding of the litsea cubeba essential oil by the medium-long chain diglyceride comprises the following steps:
(1) Preparation of a nanostructured lipid carrier based on medium-long chain diglyceride embedded litsea cubeba essential oil:
dissolving litsea cubeba essential oil in vegetable oil, adding the obtained mixed liquid oil into melted medium-long chain diglyceride, heating and stirring to obtain an oil phase; adding an emulsifying agent into a phosphate buffer solution, heating and stirring to obtain a water phase; adding the water phase into the oil phase, and carrying out high-speed shearing homogenization to obtain coarse emulsion; immediately carrying out ultrasonic treatment on the crude emulsion to obtain dispersion liquid;
(2) Cooling: and (3) immediately stirring, cooling and cooling the dispersion liquid in an ice bath after ultrasonic treatment to crystallize the lipid, thereby obtaining the nano-structure lipid carrier based on the medium-long chain diglyceride embedded litsea cubeba essential oil, namely the emulsion embedded with the litsea cubeba essential oil.
The medium-long chain diglyceride in the step (1) can be prepared by esterifying a monoglyceride containing medium-chain fatty acid and long-chain fatty acid under the catalysis of lipase, or performing transesterification of the monoglyceride containing medium-chain fatty acid and triglyceride containing long-chain fatty acid under the catalysis of lipase; or can be prepared by esterifying monoglyceride containing long-chain fatty acid and medium-chain fatty acid under lipase catalysis, or transesterifying monoglyceride containing long-chain fatty acid and triglyceride containing medium-chain fatty acid under lipase catalysis; the melting point range of the medium-long chain diglyceride is 40-70 ℃, wherein the medium-chain fatty acid is selected from one of capric acid and lauric acid, and the long-chain fatty acid is selected from one of myristic acid, palmitic acid or stearic acid.
Purifying the crude product by molecular distillation or solvent method to obtain medium-long chain diglyceride with purity not less than 50%. The molecular distillation method selects the distillation temperature of 140-200 ℃.
When the medium-long chain diglyceride is prepared by esterifying monoglyceride and fatty acid (monoglyceride and long chain fatty acid containing medium chain fatty acid or monoglyceride and medium chain fatty acid containing long chain fatty acid) under the catalysis of lipase, wherein the lipase is at least one of NoveXin 435 immobilized lipase, lipozymeCALB lipase and TLIM lipase; the mol ratio of the monoglyceride to the fatty acid is 1:3-3:1, the using amount of the lipase is 3-5wt% of the total mass of the monoglyceride and the fatty acid, the temperature of the esterification reaction is 60-70 ℃, and the reaction time is 30min-3h;
when the medium-long chain diglyceride is prepared by transesterification of monoglyceride and triglyceride (monoglyceride containing medium-chain fatty acid and triglyceride composed of long-chain fatty acid, or monoglyceride containing long-chain fatty acid and triglyceride composed of medium-chain fatty acid) under the catalysis of lipase, wherein the lipase is at least one of NoveXin 435 immobilized lipase, lipozymeCALB lipase, and TLIM lipase; the mol ratio of the monoglyceride to the triglyceride is 1:3-3:1, the using amount of lipase is 3-5wt% of the total mass of the monoglyceride and the triglyceride, the temperature of the transesterification reaction is 60-70 ℃, and the reaction time is 30min-3h.
Preferably, the vegetable oil in the step (1) is at least one of olive oil, soybean oil, corn oil, sunflower seed oil, tea seed oil, cotton seed oil, rice bran oil, peanut oil and linseed oil.
The nanostructure lipid carrier (nanostructured lipid carriers, NLCs) is used as a novel nano delivery system, can protect loaded bioactive substances from environmental factors, improves water solubility, stability and bioavailability, and enhances controlled release and targeting. The lipid matrix of NLCs is formed by mixing solid lipid and liquid lipid, so that the defects of other colloid carriers such as nanoemulsion and solid lipid nanoparticles can be overcome, and compared with Nanoemulsion (NE), the bioactive substances have weak fluidity in the solid matrix, so that the slow release can be better realized; whereas the presence of liquid oil slows down the polymorphic transition of single type lipid crystals and provides more disordered lattice to accommodate bioactive substances, increasing encapsulation efficiency compared to solid lipid nanoparticles (solid lipid nanoparticles, SLNs) with only solid matrix; other embedding systems such as polymer nanoparticles typically use organic solvents; the liposome has complex preparation process; the microemulsion has the defects of low encapsulation efficiency, poor stability and the like.
Preferably, the emulsifier in the step (1) is one of tween 80, tween 60 and tween 20; the phosphate ion concentration in the Phosphate Buffer (PBS) is 0.5-10mM, ph=6-8.
Preferably, in the step (1), the mass ratio of the oil phase to the water phase is (5:95) - (20:80), the amount of the emulsifier is 2.0-8.0% of the water phase (namely, the added amount of the emulsifier accounts for 1.6-7.6% of the total mass of the oil phase and the water phase), the amount of the medium-long chain diglyceride is 1.0-5.0% of the total mass of the emulsion, the vegetable oil is 1.0-5.0% of the total mass of the emulsion, and the litsea cubeba essential oil is 3.0-10.0% of the total mass of the emulsion. The sum of the mass percentages of the phosphate buffer solution, the diglyceride, the vegetable oil, the litsea cubeba essential oil and the emulsifier is 100 percent.
Preferably, in the step (1), the water phase and the oil phase are heated, the temperature is kept at 55-75 ℃, and volatilization of litsea cubeba essential oil is reduced while melting of medium-long chain diglyceride is ensured.
Preferably, the high-speed shearing homogenizing speed in the step (1) is 8000-15000 rpm, the homogenizing time is 2-5 minutes, and the water bath temperature in the shearing homogenizing process is 55-75 ℃.
Preferably, the power of the ultrasonic probe in the step (1) is 300-900W, the ultrasonic time is 4-10 minutes, and the water bath temperature in the ultrasonic process is 55-75 ℃.
Preferably, in the step (2), the dispersion liquid after ultrasonic treatment is placed in an ice-water bath, stirred and cooled to room temperature, and the stirring speed is 100-1000 rpm.
The invention discloses a preparation method of a nano-structure lipid carrier with a medium-long chain diglyceride as solid lipid for embedding litsea cubeba essential oil, wherein the diglyceride simultaneously has hydrophilic groups and lipophilic groups, can better reduce the interfacial tension of oil-water, has high encapsulation efficiency and high stability, improves the solubility and stability of the litsea cubeba essential oil in water, slows down the volatilization speed of the essential oil, and achieves the effects of lasting bacteriostasis and antioxidation.
The nano-structured lipid carrier based on the medium-long chain diglyceride embedded litsea cubeba essential oil prepared by the method. The nanostructure lipid carrier has light white appearance, high encapsulation efficiency, good fluidity, and lemon fragrance of litsea cubeba essential oil. The stability and biological activity of the essential oil can be protected during processing and storage, ideal slow release can be realized, the difficult problems that the litsea cubeba essential oil is difficult to dissolve in water and has high volatility and difficult utilization as oil are solved, and the broad-spectrum antibacterial property and the antioxidant capacity of the litsea cubeba essential oil are fully exerted. The properties of the lipid carrier embedded with different bioactive compounds can be regulated and controlled by changing the preparation conditions such as the medium-long chain diglyceride/vegetable oil ratio, the vegetable oil/essential oil ratio, the type and the content of the emulsifier and the like in the lipid carrier with the nano structure.
The particle size of the nano-structured lipid carrier based on the medium-long chain diglyceride embedded litsea cubeba essential oil is 100-300nm, and the encapsulation rate is 92-98%.
Application of nano-structured lipid carrier based on long-chain diglyceride embedded litsea cubeba essential oil in fruit and vegetable fresh-keeping comprises the following steps:
(1) Preparing a bacteriostatic paper box: uniformly coating the emulsion embedded with litsea cubeba essential oil on corrugated paper or white paperboard by using a spraying mode or a coating rod, naturally air-drying or drying in an electrothermal constant-temperature blast drying oven, and then folding into a bacteriostatic paper box;
(2) The fruits and vegetables are placed in a bacteriostatic paper box and stored in a refrigerator.
Compared with the prior art, the invention has the following advantages and effects:
(1) The nano-structure lipid carrier prepared by embedding litsea cubeba essential oil with the medium-long chain diglyceride solves the difficult problems that the litsea cubeba essential oil is not easy to dissolve in water and is difficult to use due to high volatility, realizes high-efficiency embedding and sustained release of the litsea cubeba essential oil, and fully exerts the broad-spectrum antibacterial property and oxidation resistance of the litsea cubeba essential oil. The litsea cubeba essential oil is wrapped in the water-based nano-structured lipid carrier, and can be used for preserving fruits and vegetables in a soaking and spraying mode, and the high stability of the nano-structured lipid carrier is also beneficial to the application of fruits and vegetables in storage and cold chain transportation, so that the nano-structured lipid carrier has a wide application prospect in the field of preserving fruits and vegetables.
(2) The invention has simple process, does not relate to the application of organic reagents, has simple ultrasonic treatment operation and environmental protection, can carry out rapid continuous preparation, can prepare emulsion embedding different bioactive compounds by simply regulating and controlling the proportion of medium-long chain diglyceride/vegetable oil, the proportion of vegetable oil/essential oil, the type and content of emulsifying agent and the like in the nano-structure lipid carrier based on the medium-long chain diglyceride embedded litsea cubeba essential oil, and has better industrialized application value.
(3) The medium-long chain diglyceride used in the invention has excellent molecular amphipathy, high melting point, good emulsifying property and difficult occurrence of crystal form conversion in the storage process. The prepared nano-structured lipid carrier based on the litsea cubeba essential oil embedded by the medium-long chain diglyceride has light white appearance, good fluidity, moderate viscosity, high encapsulation efficiency and high stability, can be infinitely diluted by water or phosphoric acid buffer solution, is stable at low temperature, and has outstanding advantages compared with the traditional emulsion and solid lipid nanoparticles which are easy to generate crystal form transformation in the storage process and have lower encapsulation efficiency.
(4) The size of the paper box tray can be made by beating according to the requirements, has stronger flexibility, and has better preference compared with the traditional common paper box treated by chemical preservative, such as coated corrugated paper boxes, paper boards and the like which are specially used for food packaging. The green and safe natural plant essential oil with broad-spectrum antibacterial effect is added into the food package, so that the food package can be used for relieving the putrefaction of fruits, and can also reduce the residue of chemical preservative, so that the packaging material is healthier and more environment-friendly.
Drawings
Fig. 1 is a schematic flow chart of the present invention.
Fig. 2 is a particle size distribution diagram, a microscopic image and a solid image of a nanostructured lipid carrier based on the medium-long chain diglyceride embedded litsea cubeba essential oil of example 1.
Fig. 3 shows the encapsulation efficiency of example 1, example 2, example 3, comparative example 1 and comparative example 2.
Fig. 4 is a graph showing the effect of example 1, example 2, example 3, comparative example 1 and comparative example 2 on the active oxygen scavenging ability.
Fig. 5 shows the morphology changes during storage of blueberries of example 1, example 2, example 3, comparative example 1, comparative example 2 and control group.
Fig. 6 shows the variation of the rotting rate of blueberries of example 1, example 2, example 3, comparative example 1, comparative example 2 and control group.
Fig. 7 is a variation of the blueberry weight loss rate of example 1, example 2, example 3, comparative example 1, comparative example 2 and control group.
Fig. 8 is a graph showing the variation in hardness of blueberries of example 1, example 2, example 3, comparative example 1, comparative example 2 and control group.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but embodiments of the present invention are not limited thereto. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
The reagents used in the examples are commercially available as usual unless otherwise specified.
Example 1:
the preparation method of the nano-structured lipid carrier based on the embedding of the litsea cubeba essential oil by the medium-long chain diglyceride comprises the steps that the emulsion is formed by compounding a water phase and an oil phase containing the litsea cubeba essential oil, wherein the mass ratio of the water phase is 90%, and the mass ratio of the oil phase is 10%; wherein the aqueous phase comprises 5.4% tween 80 and 84.6% 5mM phosphate buffer; the oil phase comprises 2% of medium-long chain diglyceride, 3% of olive oil and 5% of litsea cubeba essential oil, and the preparation steps are as follows:
(1) Preparing and purifying a medium-long chain diglyceride raw material: mixing lauric acid and monoglyceride stearate according to a molar ratio of 3:1, carrying out vacuum catalytic reaction at 60 ℃ for 30min, wherein the catalyst is NoveXin 435 immobilized lipase, the addition amount is 5% of the total mass of a substrate, purifying the reacted crude product by molecular distillation at 180 ℃ to obtain purified diglyceride, and detecting by adopting gas chromatography, wherein the proportion of medium-long chain diglyceride is more than 60%.
(2) Heating and stirring the medium-long chain diglyceride containing lauric acid and stearic acid at 70 ℃ to completely melt, dissolving litsea cubeba essential oil in olive oil, and adding the mixture into the melted medium-long chain diglyceride to obtain an oil phase; adding Tween 80 into 5mM phosphate buffer solution, preheating at 70deg.C, and stirring to obtain water phase; adding the water phase into the oil phase, shearing and homogenizing the mixed solution at a high speed of 12000rpm, and preparing coarse emulsion at a water bath temperature of 70 ℃; ultrasonic treatment is carried out for 6 minutes under the power of 540W to obtain diglyceride dispersion liquid embedding essential oil;
(2) And (3) immediately using a magnetic stirrer at a rotating speed of 300rpm to stir and cool the dispersion liquid obtained in the step (1) to obtain the nano-structured lipid carrier of the medium-long chain diglyceride embedded litsea cubeba essential oil.
The application of the nano-structured lipid carrier based on the medium-long chain diglyceride embedded litsea cubeba essential oil obtained by the preparation method in the antibacterial paper box comprises the following steps of:
(1) Preparing a bacteriostatic paper box: the emulsion embedded with litsea cubeba essential oil is 80g/m 2 Uniformly coating on corrugated paper, naturally air-drying, and folding to obtain the invented antibacterial paper box.
(2) And placing the blueberries in a bacteriostatic paper box, and storing in a refrigerator.
Preferably, the size of the bacteriostatic paper box tray can be designed according to actual conditions; the fresh-keeping temperature of the refrigerator is 4-8 ℃.
Example 2:
the preparation method of the nano-structured lipid carrier based on the litsea cubeba essential oil embedded by the medium-long chain diglyceride is the same as that of the embodiment 1, wherein the nano-structured lipid carrier is prepared by compounding an aqueous phase and an oil phase containing the litsea cubeba essential oil, the mass ratio of the aqueous phase is 90%, and the mass ratio of the oil phase is 10%; wherein the aqueous phase is 7.2% tween 20 and 82.8%5mm phosphate buffer; the oil phase was 2.5% medium-long chain diglyceride containing lauric acid and stearic acid, 2.5% soybean oil, and 5% litsea cubeba essential oil, and the preparation procedure was as in example 1.
Example 3:
the preparation method of the nano-structured lipid carrier based on the litsea cubeba essential oil embedded by the medium-long chain diglyceride is the same as that of the embodiment 1, wherein the nano-structured lipid carrier is prepared by compounding an aqueous phase and an oil phase containing the litsea cubeba essential oil, the mass ratio of the aqueous phase is 85%, and the mass ratio of the oil phase is 15%; wherein the aqueous phase is 7.2% tween 20 and 77.8%5mm phosphate buffer; the oil phase comprised 5% lauric acid, stearic acid medium chain diglyceride, 5% soybean oil and 5% litsea cubeba essential oil was prepared in the same manner as in example 1.
The test method comprises the following steps:
the particle size distribution and microscopic morphology of the nanostructured lipid carrier were analyzed using a dynamic light scattering analyzer and a transmission electron microscope, respectively.
Encapsulation efficiency measurement: mixing the emulsion with 50% ethanol according to a volume ratio of 1: 7% and centrifuging at 4000rpm for 5 min in an ultrafiltration tube to separate the free essential oil from the ultrafiltration membrane, collecting the filtrate, and diluting with 50% ethanol. Adding 1mL chloroform to 1mL emulsion and shaking sufficiently to destroy carrier and release encapsulated essential oil, centrifuging at 10000 for 10 min, collecting chloroform layer containing free and embedded essential oil, diluting with 50% ethanol, and measuring lambda with ultraviolet-visible spectrophotometer max Measured at =238 nm. Drawing a standard curve y= 0.00002881674x-0.00000056298 by using litsea cubeba essential oil with concentration range of 0.00001-0.00005 mg/mL, and calculating essential oil content, wherein a correlation coefficient R 2 =0.987, y is the concentration of litsea cubeba essential oil (mg/mL), x is the absorbance (a) value of the solution at 238nm, and the encapsulation efficiency is calculated using the following formula:
DPPH radical scavenging Rate determination: preparing 0.1-1.0 mu L/mL of sample methanol solution, mixing 500 mu L of the sample methanol solution with 3.0mL of 0.06mM DPPH methanol solution, and measuring absorbance at 517nm, wherein the methanol solution without the sample is used as a blank control. The number of replicates was 3 and the radical scavenging rate was according to the following formula:
the IC is calculated by plotting the linear relation of the concentration and the clearance of the sample 50 I.e. the concentration of nanostructured lipid carrier dispersion required to scavenge 50% of free radicals.
Wherein A is Sample of Absorbance at 517nm for the sample; a is that Blank space The absorbance at 517nm was used as a blank.
Blueberry decay rate evaluation: the rate of decay is evaluated based on the degree of softening of the fruit, the leakage of juice caused by decay, and the degree of mold infection. According to the spoilage, the waste water is classified into 3 grades, namely 0 grade: no obvious change on the surface; stage 1: obvious softening phenomenon occurs; 2 stages: the softening is serious, and the juice leakage phenomenon occurs; 3 stages: mildew infection appears on the surface.
The decay rate is calculated as follows:
blueberry weight loss rate evaluation:
blueberry hardness measurement: and (3) performing puncture measurement by adopting a texture analyzer and aligning a 2mm probe with the equatorial region of the blueberry. The penetration depth was 4mm, the speed was 1.5mm/s, the trigger force was 7.0g, and 15 measurements were made per treatment in g.
Example 4: effect of emulsifier type and concentration on nanostructured lipid Carrier Properties based on Medium-Long chain diglyceride-embedded Litsea cubeba essential oil
In this example, 5mM phosphate buffer was used as the water phase, and the oil phase was: medium-long chain diglycerides (MLCD) containing lauric acid, stearic acid (the medium-long chain diglycerides were prepared and purified as in example 1), and a mixed liquid oil having an olive oil to essential oil quality ratio of 1:1. The effect of different concentrations of emulsifier (tween 20 or tween 80) on the particle size, potential and Dispersibility Index (PDI) of the nanoemulsion was measured by a dynamic light scattering analyzer, and specific parameters are shown in table 1, wherein the total content of essential oil and olive oil is 5% of the total mass of the water phase and oil phase, the content of MLCD is 5% of the total mass of the water phase and oil phase, and the addition amount of tween 80 is the ratio of the total mass of the water phase and oil phase.
TABLE 1 nanostructured lipid Carrier Properties based on Medium-Long chain diglyceride-embedded Litsea cubeba essential oil at different emulsifier types and addition levels
Different emulsifier types and concentrations can form emulsions of different particle sizes (Table 1). Tween-series emulsifiers all assisted in forming smaller size and uniformly dispersed nanostructured lipid carriers based on medium-long chain diglycerides, formulated with 5.4% tween 80 in the following examples.
Example 5: effect of olive oil to essential oil ratio on nanostructure lipid Carrier Properties
In this example, the aqueous phase is 0.5mM phosphate buffer, the emulsifier is Tween 80, and the oil phase comprises: medium-long chain diglyceride (MLCD) (the preparation and purification of the medium-long chain diglyceride is the same as in example 1), olive oil, litsea cubeba essential oil. The effect of different ratios of olive oil to essential oil on the particle size, potential and PDI of the nanoemulsion was chosen, as shown in table 2. Wherein the total content of essential oil and olive oil is 5% of the total mass of the water phase and oil phase, the content of MLCD is 5% of the total mass of the water phase and oil phase, and the addition amount of Tween 80 is 5.4% of the total mass of the water phase and oil phase.
TABLE 2 Effect of olive oil to essential oil ratio on nanostructured lipid Carrier Properties
The average particle size of the emulsion was more distinct at different ratios of olive oil to essential oil (table 2). When the olive oil to essential oil quality ratio is 1:1, the nanoemulsion has the smallest particle size and good dispersibility.
Example 6: effect of Medium-Long chain diglyceride to olive oil ratio on nanostructured lipid Carrier Properties based on Medium-Long chain diglyceride-embedded Litsea cubeba essential oil
In this example, 5mM phosphate buffer solution is used as the water phase, tween 80 is used as the emulsifier, and the oil phase comprises: medium-long chain diglyceride (MLCD) containing lauric acid and stearic acid (the preparation and purification of the medium-long chain diglyceride are the same as in example 1), olive oil and litsea cubeba essential oil. The influence of different medium-long chain diglyceride and olive oil ratios on the particle size, potential and PDI of the nanoemulsion is analyzed, and the specific reference is shown in table 2, wherein the total content of MLCD and olive oil accounts for 5% of the total mass of the water phase and the oil phase, and the addition amounts of essential oil and tween 80 refer to the ratio of the mass of the essential oil to the total mass of the water phase and the oil phase.
TABLE 3 influence of the ratio of medium-long chain diglycerides to olive oil on the properties of nanostructured lipid carriers based on the embedding of Litsea cubeba essential oil with medium-long chain diglycerides
The emulsion particle sizes formed at different diglyceride to olive oil ratios were different (table 3). When the mass ratio of the medium-long chain diglyceride to the olive oil is 2:3, the formed nano emulsion has the smallest particle size and is uniformly dispersed.
Comparative example 1
(1) Dissolving fructus Litseae essential oil in oleum Olivarum, and stirring, wherein the weight ratio of oleum Olivarum to fructus Litseae essential oil is 1:1, obtaining an oil phase; adding Tween 80 into 5mM phosphate buffer solution, and stirring to obtain water phase; the aqueous phase is added into the oil phase, the mixed solution is sheared and homogenized at a high speed at 15000rpm, and then ultrasonic is carried out for 6 minutes under the power of 540W, so that the traditional nano emulsion embedding the essential oil is obtained.
The nano emulsion embedded with litsea cubeba essential oil comprises 5% of olive oil, 5% of litsea cubeba essential oil, 5.4% of tween 80 and 84.6% of phosphate buffer solution.
Comparative example 2
(1) Heating and stirring medium-long chain diglyceride containing lauric acid and stearic acid (the preparation and purification of the medium-long chain diglyceride are the same as in example 1) at 60 ℃ to completely melt the medium-long chain diglyceride, adding litsea cubeba essential oil into the medium-long chain diglyceride (the mass ratio is 1:1), and heating and stirring at 60 ℃ to obtain an oil phase; adding Tween 80 into 5mM phosphate buffer solution, preheating at 60deg.C, and stirring to obtain water phase; adding the water phase into the oil phase, and shearing and homogenizing the mixed solution at a high speed of 15000rpm to obtain a coarse emulsion; ultrasonic treatment is carried out for 6 minutes under the power of 540W to obtain diglyceride dispersion liquid embedding essential oil;
(2) And (3) immediately using a magnetic stirrer at a rotating speed of 300rpm to stir and cool the dispersion liquid obtained in the step (1) to obtain solid lipid nanoparticles embedded with litsea cubeba essential oil.
The solid lipid nanoparticle embedded with litsea cubeba essential oil comprises 5% of medium-long chain diglyceride, 5% of litsea cubeba essential oil, 5.4% of tween-80 and 84.6% of phosphate buffer solution.
The emulsion properties and blueberry storage process properties were measured as shown in tables 4 and 5:
TABLE 4 emulsion Properties
Analysis: as can be seen from Table 4 above, the NLC prepared in example 1 has a light white appearance, uniform dispersion, good fluidity, fresh lemon flavor, particle size of 181.21nm, PDI of 0.153, encapsulation efficiency of 97.69%, DPPH removal rate IC 50 At 0.76 μl/mL, and after 60 days of storage at 4 ℃ there was no significant change in both particle size and PDI, the encapsulation efficiency was slightly reduced to 93.84%, showing higher entrapment rate, better stability and stronger free radical scavenging ability than comparative examples 1, 2.
TABLE 5 blueberry storage process property measurements
Analysis: as can be seen from Table 5 above, all of the examples and comparative examples had some retardation effect on the spoilage of blueberries compared to the control group without any treatment, the blueberries treated in example 1 had spoilage rates of 0%, 0.83%, 3.33% and 5.83% on days 1, 4, 8 and 12, weight loss rates of 0.29%, 2.12%, 6.05% and 10.69% and hardness of 279g, 267g, 255g and 241g, respectively. The blueberries have no obvious change after 4 days of storage, and a small part of blueberry fruits begin to soften after 8 days.
After 12 days of storage, the untreated blueberry rotting rate was 2.79 times that of the NLC treated group, the weight loss percentage was 1.49 times, and the softening degree of the blueberry fruits kept fresh by NLC was minimal. Compared with the nanoemulsions of examples 2 and 3 and comparative example 1 and the solid lipid nanoparticle of comparative example 2, the NLC prepared in example 1 has better antioxidation and bacteriostasis capacity and has the best fresh-keeping effect on blueberry fruits.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (7)
1. A method for preparing a nanostructured lipid carrier, comprising the steps of:
(1) Dissolving litsea cubeba essential oil in olive oil, adding the obtained mixed oil into melted medium-long chain diglyceride, heating and stirring to obtain an oil phase; adding Tween 80 into phosphate buffer solution, heating and stirring to obtain water phase; adding the water phase into the oil phase, and carrying out high-speed shearing homogenization to obtain coarse emulsion; immediately carrying out ultrasonic treatment on the crude emulsion to obtain dispersion liquid;
(2) After ultrasonic treatment, immediately stirring and cooling the dispersion liquid in an ice bath to obtain a nano-structure lipid carrier based on the medium-long chain diglyceride embedded litsea cubeba essential oil;
the two corresponding fatty acids after the medium-long chain diglyceride is completely hydrolyzed in the step (1) are medium-chain fatty acid and long-chain fatty acid, wherein the medium-chain fatty acid is lauric acid, and the long-chain fatty acid is stearic acid; the melting point range of the medium-long chain diglyceride in the step (1) is 40-70 ℃, and the purity is not lower than 50%;
in the step (1), the mass ratio of the oil phase to the water phase is 10:90, wherein the amount of Tween 80 is 5.4% of the water phase, the amount of medium-long chain diglyceride is 2.0% of the total mass of the emulsion, the amount of olive oil is 3.0% of the total mass of the emulsion, and the amount of litsea cubeba essential oil is 5.0% of the total mass of the emulsion.
2. The method for preparing a nanostructured lipid carrier according to claim 1, wherein,
the medium-long chain diglyceride is prepared by esterifying a monoglyceride containing medium-chain fatty acid and long-chain fatty acid under the catalysis of lipase, or is prepared by transesterifying a monoglyceride containing medium-chain fatty acid and a triglyceride containing long-chain fatty acid under the catalysis of lipase; or is prepared by esterifying monoglyceride containing long-chain fatty acid and medium-chain fatty acid under lipase catalysis, or is prepared by transesterifying monoglyceride containing long-chain fatty acid and triglyceride containing medium-chain fatty acid under lipase catalysis; wherein the medium chain fatty acid is lauric acid, and the long chain fatty acid is stearic acid.
3. The method for preparing a nanostructured lipid carrier according to claim 1, wherein the heating temperature of the aqueous phase and the oil phase in the step (1) is kept at 55-75 ℃;
the high-speed shearing homogenizing speed in the step (1) is 8000-15000 rpm, and the homogenizing time is 2-5 minutes;
the power of the ultrasonic probe in the step (1) is 300-900W, and the ultrasonic time is 4-10 minutes;
and (2) placing the dispersion liquid subjected to ultrasonic treatment in an ice bath, stirring and cooling to room temperature, wherein the stirring speed is 100-1000 rpm.
4. A nanostructured lipid carrier, characterized in that it is obtainable by the method according to any one of claims 1-3.
5. The use of the nanostructured lipid carrier according to claim 4 for preserving fruits and vegetables.
6. The use of the nanostructured lipid carrier according to claim 4 for preserving blueberries.
7. Use of a nanostructured lipid carrier according to claim 5, characterized in that it comprises the steps of:
(1) Preparing a bacteriostatic paper box: spraying the nano-structure lipid carrier embedded with litsea cubeba essential oil on corrugated paper or white cardboard, naturally air-drying or drying in an electrothermal constant-temperature blast drying oven, and then folding into a bacteriostatic paper box;
(2) The fruits and vegetables are placed in a bacteriostatic paper box and stored in a refrigerator.
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| PCT/CN2022/106735 WO2023206830A1 (en) | 2022-04-24 | 2022-07-20 | Preparation and application of nanostructured lipid carrier based on medium- and long-chain diglyceride embedded litsea cubeba essential oil |
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| CN114698688B (en) * | 2022-04-24 | 2023-09-26 | 暨南大学 | Preparation and application of nanostructure lipid carrier |
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