CN115843864A - Composite volatile oil ethosome and preparation method thereof - Google Patents
Composite volatile oil ethosome and preparation method thereof Download PDFInfo
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- CN115843864A CN115843864A CN202211575233.5A CN202211575233A CN115843864A CN 115843864 A CN115843864 A CN 115843864A CN 202211575233 A CN202211575233 A CN 202211575233A CN 115843864 A CN115843864 A CN 115843864A
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- volatile oil
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- 239000000341 volatile oil Substances 0.000 title claims abstract description 92
- 239000002131 composite material Substances 0.000 title claims abstract description 49
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 45
- 239000004094 surface-active agent Substances 0.000 claims abstract description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 23
- 239000012528 membrane Substances 0.000 claims abstract description 13
- 238000002156 mixing Methods 0.000 claims abstract description 12
- 150000003904 phospholipids Chemical class 0.000 claims abstract description 12
- 239000002994 raw material Substances 0.000 claims abstract description 10
- 150000001875 compounds Chemical class 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims abstract description 8
- 238000000265 homogenisation Methods 0.000 claims abstract description 4
- 239000012071 phase Substances 0.000 claims description 13
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- 238000003756 stirring Methods 0.000 claims description 8
- 239000008346 aqueous phase Substances 0.000 claims description 7
- 229930182558 Sterol Natural products 0.000 claims description 6
- 235000012000 cholesterol Nutrition 0.000 claims description 6
- 235000003702 sterols Nutrition 0.000 claims description 6
- PNQGEMVOEPSFES-ZPGRZCPFSA-N (2S)-5-[[(2R)-1-(carboxymethylamino)-3-hexadecanoylsulfanyl-1-oxopropan-2-yl]amino]-2-(hexadecanoylamino)-5-oxopentanoic acid Chemical compound CCCCCCCCCCCCCCCC(=O)N[C@H](C(O)=O)CCC(=O)N[C@H](C(=O)NCC(O)=O)CSC(=O)CCCCCCCCCCCCCCC PNQGEMVOEPSFES-ZPGRZCPFSA-N 0.000 claims description 5
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- LGJMUZUPVCAVPU-UHFFFAOYSA-N beta-Sitostanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 LGJMUZUPVCAVPU-UHFFFAOYSA-N 0.000 claims description 4
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- 125000005257 alkyl acyl group Chemical group 0.000 claims description 3
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 3
- ASWBNKHCZGQVJV-UHFFFAOYSA-N (3-hexadecanoyloxy-2-hydroxypropyl) 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(O)COP([O-])(=O)OCC[N+](C)(C)C ASWBNKHCZGQVJV-UHFFFAOYSA-N 0.000 claims description 2
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 claims description 2
- PZNPLUBHRSSFHT-RRHRGVEJSA-N 1-hexadecanoyl-2-octadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)O[C@@H](COP([O-])(=O)OCC[N+](C)(C)C)COC(=O)CCCCCCCCCCCCCCC PZNPLUBHRSSFHT-RRHRGVEJSA-N 0.000 claims description 2
- OQMZNAMGEHIHNN-UHFFFAOYSA-N 7-Dehydrostigmasterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(CC)C(C)C)CCC33)C)C3=CC=C21 OQMZNAMGEHIHNN-UHFFFAOYSA-N 0.000 claims description 2
- OILXMJHPFNGGTO-NRHJOKMGSA-N Brassicasterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@](C)([C@H]([C@@H](/C=C/[C@H](C(C)C)C)C)CC4)CC3)CC=2)CC1 OILXMJHPFNGGTO-NRHJOKMGSA-N 0.000 claims description 2
- SGNBVLSWZMBQTH-FGAXOLDCSA-N Campesterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]([C@H](CC[C@H](C(C)C)C)C)CC4)CC3)CC=2)CC1 SGNBVLSWZMBQTH-FGAXOLDCSA-N 0.000 claims description 2
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 claims description 2
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- BTEISVKTSQLKST-UHFFFAOYSA-N Haliclonasterol Natural products CC(C=CC(C)C(C)(C)C)C1CCC2C3=CC=C4CC(O)CCC4(C)C3CCC12C BTEISVKTSQLKST-UHFFFAOYSA-N 0.000 claims description 2
- QZWIYJJEFUABFJ-IFJPITGXSA-N N[C@@H](CCCCN)C(=O)O.C(CCCCCCCCCCC)(=O)NN[C@@H](CCC(N)=O)C(=O)O.C(CCCCCCCCCCC)(=O)NN[C@@H](CCC(N)=O)C(=O)O.[Na] Chemical compound N[C@@H](CCCCN)C(=O)O.C(CCCCCCCCCCC)(=O)NN[C@@H](CCC(N)=O)C(=O)O.C(CCCCCCCCCCC)(=O)NN[C@@H](CCC(N)=O)C(=O)O.[Na] QZWIYJJEFUABFJ-IFJPITGXSA-N 0.000 claims description 2
- OILXMJHPFNGGTO-ZRUUVFCLSA-N UNPD197407 Natural products C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)C=C[C@H](C)C(C)C)[C@@]1(C)CC2 OILXMJHPFNGGTO-ZRUUVFCLSA-N 0.000 claims description 2
- HZYXFRGVBOPPNZ-UHFFFAOYSA-N UNPD88870 Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)=CCC(CC)C(C)C)C1(C)CC2 HZYXFRGVBOPPNZ-UHFFFAOYSA-N 0.000 claims description 2
- ATBOMIWRCZXYSZ-XZBBILGWSA-N [1-[2,3-dihydroxypropoxy(hydroxy)phosphoryl]oxy-3-hexadecanoyloxypropan-2-yl] (9e,12e)-octadeca-9,12-dienoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCC\C=C\C\C=C\CCCCC ATBOMIWRCZXYSZ-XZBBILGWSA-N 0.000 claims description 2
- AWUCVROLDVIAJX-UHFFFAOYSA-N alpha-glycerophosphate Natural products OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 claims description 2
- 229940076810 beta sitosterol Drugs 0.000 claims description 2
- NJKOMDUNNDKEAI-UHFFFAOYSA-N beta-sitosterol Natural products CCC(CCC(C)C1CCC2(C)C3CC=C4CC(O)CCC4C3CCC12C)C(C)C NJKOMDUNNDKEAI-UHFFFAOYSA-N 0.000 claims description 2
- OILXMJHPFNGGTO-ZAUYPBDWSA-N brassicasterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)/C=C/[C@H](C)C(C)C)[C@@]1(C)CC2 OILXMJHPFNGGTO-ZAUYPBDWSA-N 0.000 claims description 2
- 235000004420 brassicasterol Nutrition 0.000 claims description 2
- SGNBVLSWZMBQTH-PODYLUTMSA-N campesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CC[C@@H](C)C(C)C)[C@@]1(C)CC2 SGNBVLSWZMBQTH-PODYLUTMSA-N 0.000 claims description 2
- 235000000431 campesterol Nutrition 0.000 claims description 2
- FRKBLBQTSTUKOV-UHFFFAOYSA-N diphosphatidyl glycerol Natural products OP(O)(=O)OCC(OP(O)(O)=O)COP(O)(O)=O FRKBLBQTSTUKOV-UHFFFAOYSA-N 0.000 claims description 2
- 239000008344 egg yolk phospholipid Substances 0.000 claims description 2
- 150000002632 lipids Chemical class 0.000 claims description 2
- 125000001095 phosphatidyl group Chemical group 0.000 claims description 2
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 claims description 2
- 150000008104 phosphatidylethanolamines Chemical class 0.000 claims description 2
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- 238000007789 sealing Methods 0.000 claims description 2
- KZJWDPNRJALLNS-VJSFXXLFSA-N sitosterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CC[C@@H](CC)C(C)C)[C@@]1(C)CC2 KZJWDPNRJALLNS-VJSFXXLFSA-N 0.000 claims description 2
- 229950005143 sitosterol Drugs 0.000 claims description 2
- NLQLSVXGSXCXFE-UHFFFAOYSA-N sitosterol Natural products CC=C(/CCC(C)C1CC2C3=CCC4C(C)C(O)CCC4(C)C3CCC2(C)C1)C(C)C NLQLSVXGSXCXFE-UHFFFAOYSA-N 0.000 claims description 2
- 239000008347 soybean phospholipid Substances 0.000 claims description 2
- 150000003432 sterols Chemical class 0.000 claims description 2
- HCXVJBMSMIARIN-PHZDYDNGSA-N stigmasterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)/C=C/[C@@H](CC)C(C)C)[C@@]1(C)CC2 HCXVJBMSMIARIN-PHZDYDNGSA-N 0.000 claims description 2
- 235000016831 stigmasterol Nutrition 0.000 claims description 2
- 229940032091 stigmasterol Drugs 0.000 claims description 2
- BFDNMXAIBMJLBB-UHFFFAOYSA-N stigmasterol Natural products CCC(C=CC(C)C1CCCC2C3CC=C4CC(O)CCC4(C)C3CCC12C)C(C)C BFDNMXAIBMJLBB-UHFFFAOYSA-N 0.000 claims description 2
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Abstract
The invention provides a compound volatile oil ethosome and a preparation method thereof, and relates to the technical field of food fresh-keeping and bacteriostasis. A composite volatile oil ethosome comprises the following raw materials in percentage by weight except water: 0.1-3% of compound volatile oil, 0.5-5% of phospholipid, 0.01-1% of membrane fluidity regulator, 10-20% of ethanol and 0.1-0.4% of surfactant. The preparation method of the composite volatile oil ethosome comprises the following steps: and carrying out first mixing and homogenization on the raw materials and water to obtain the composite volatile oil ethosome. The composite volatile oil ethosome composition with high encapsulation efficiency and low ethanol content provided by the invention has the characteristics of good antibacterial effect, long slow release time, low alcohol irritation, excellent stability and the like, is green and environment-friendly in preparation raw materials, simple in process, controllable in condition and has wide industrial application prospect.
Description
Technical Field
The invention belongs to the technical field of food fresh-keeping and bacteriostasis, and particularly relates to a compound volatile oil ethosome and a preparation method thereof.
Background
Fresh fruits and vegetables are susceptible to contamination from pathogens at any stage from pre-harvest to post-harvest storage and transportation. In the prior art, methods for controlling fruit pathogenic bacteria mainly comprise chemical prevention and control, physical prevention and control, biological prevention and control, disease-resistant varieties and the like. The low-temperature storage can inhibit the growth of pathogenic bacteria or slow down the growth, and can reduce the enzyme activity of the fruits, thereby delaying the senescence of the fruits, effectively controlling the rot of the fruits and prolonging the storage period. But the problems of high cost and short period exist because of various limitations of physical fresh-keeping, and the fresh-keeping effect and time are not ideal. Chemical control is the main method for controlling postharvest diseases of fruits and vegetables. To prevent or reduce fruit spoilage by pathogenic microorganisms, a simple and effective method is to use antimicrobial agents. The application of chemical pesticides such as difenoconazole, iprodione, tioconazole, carbendazim, thiophanate-methyl and the like is an important means for controlling pathogenic bacteria of picked fruits. However, most of the chemical bactericides or pesticide residues have toxic and side effects, easily cause environmental pollution, destroy the ecological system and poison people and livestock. In recent years, scholars at home and abroad pay attention to the replacement of chemical bactericides by some safe and nontoxic substances, including phenols, chitosan, plant volatile oil, sodium sorbate and the like. The volatile oil treatment is used as a natural bacteriostatic agent which is safe to human bodies and friendly to the environment, and has great potential for replacing the traditional chemical preservative in the food industry. However, the volatile oil has the defects of easy volatilization, low water solubility, easy oxidation when meeting light, unstable heat and the like, and the wide application of the plant volatile oil is seriously restricted. Embedding the plant volatile oil can improve the stability of the plant volatile oil, reduce the chemical degradability, stability and biological activity of the plant volatile oil, and solve the problems in the application of the plant volatile oil to a certain extent. The nanometer ethosome is used as a mode of embedding technology liposome, and can effectively improve the water solubility and the chemical stability of the plant volatile oil. However, the ethosomes directly applied to the surface of the food as the preservative have low encapsulation efficiency and high ethanol content, and the ethanol content is usually as high as 40%, so that the putrefaction of the preserved substances can be accelerated. In addition, too high ethanol promotes evaporation of water on the surface of the preserved object, so that the surface is dried, and therefore, the ethosome containing too high alcohol content is not suitable for being used as a food preservative.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a composite volatile oil ethosome and a preparation method thereof, and aims to solve the problems.
In order to realize the purpose, the invention adopts the following technical scheme:
the composite volatile oil ethosome comprises the following raw materials in percentage by weight except water: 0.1-3% of compound volatile oil, 0.5-5% of phospholipid, 0.01-1% of membrane fluidity regulator, 10-20% of ethanol and 0.1-0.4% of surfactant.
Optionally, the composite volatile oil is selected from at least two of the following volatile oils: <xnotran> , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , D- , , , , , , , . </xnotran>
Volatile oils are a class of volatile oily components present in plants that have an aromatic odor and are water-immiscible and distill with water vapor. The composite volatile oil as a main functional component influences the overall preservative and fresh-keeping performance of the system, and provides high functionality for the preservative and fresh-keeping of the ethosome. Preferably, the weight percentage of the compound volatile oil in the raw materials is 0.4-1%. Experiments prove that when the mass ratio of the litsea cubeba oil to the tsaoko amomum fruit oil in the composite volatile oil is 7.
Optionally, the phospholipid comprises a natural phospholipid and/or a synthetic lipid;
preferably, the natural phospholipids include one or more of soy phosphatidylcholine, egg lecithin, soy lecithin, lysolecithin, sphingomyelin, phosphatidyl, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, diphosphatidylglycerol, and plasmalogen.
Preferably, the weight percentage of the phospholipids in the composite volatile oil ethosome raw material is 1-3%.
Optionally, the membrane fluidity modifier comprises a phytosterol and/or cholesterol;
preferably, the phytosterols comprise one or more of 4-methyl free sterol, 4-methyl sterol, 4' -dimethyl sterol, BETA-sitosterol, stigmasterol, campesterol and brassicasterol.
When phytosterols and/or cholesterol are added as membrane fluidity modifiers, more stable formation of ethosomes is facilitated. Cholesterol or phytosterols become wedged between phospholipids, making the membrane structure more rigid and thus contributing to the formation of a stable phospholipid membrane. Preferably, the weight percentage of the cholesterol is 0.01-0.5 percent, and the weight percentage of the phytosterol is 0.5-1.0 percent; more preferably, the weight percentage of the cholesterol is 0.1-0.2%. The appropriate amount of membrane fluidity regulator stabilizes the phospholipid membrane, which contributes to particle stabilization.
Optionally, the surfactant is a linear alkyl acyl peptide structure surfactant;
preferably, the surfactant comprises sodium di (lauramidoglutamine) lysine and/or dipalmitoyl glutathione.
The addition of the surfactant can effectively improve the deformability of the alcohol vesicle membrane and also help to inhibit particle aggregation, thereby effectively improving the stability of the alcohol body. The O/W surfactants commonly used in the preparation of liposomes and ethosomes are poloxamer-188 and tween-80, but the ethosomes prepared therefrom are not sufficiently stable. The linear alkyl acyl peptide structure surfactant adopted by the invention is of pure plant source and has a ceramide-like structure, so that the stability of an ethosome is effectively improved, and the using amount of ethanol is reduced. Preferably, the weight percentage of the sodium bis (lauramide glutamine) lysine is 0.1-0.2%, and the weight percentage of the dipalmitoyl glutathione is 0.3-0.4%.
The invention also provides a preparation method of the composite volatile oil ethosome, which comprises the following steps: and carrying out first mixing and homogenization on the raw materials and water to obtain the composite volatile oil ethosome.
Optionally, the first mixing specifically is: dissolving the surfactant in water to form an aqueous phase; dissolving said phospholipid, said membrane fluidity modifier, and said complex volatile oil in said ethanol to form an alcohol phase;
second mixing the aqueous phase and the alcohol phase.
Optionally, the second mixing comprises dropping the aqueous phase into the alcohol phase at a rate of 1 to 5mL/min under insulated, sealed conditions.
Further optionally, the second mixing is performed under a stirring state, wherein a stirring rate at the time of the dropping is 300 to 800rpm, and a stirring rate after the completion of the dropping is 500 to 800rpm.
Optionally, the homogenizing rotation speed is 6000-10000 rpm, and the time is 5-10 min.
The composite volatile oil ethosome provided by the invention is a non-thermodynamic stable system, namely a nano ethosome, which is formed by mixing active substance composite volatile oil, a surfactant, phospholipid, water and the like in different proportions, compared with non-nano emulsified volatile oil, the nano ethosome can improve the water solubility of the volatile oil, can also enable oil drops of the volatile oil to reach the nano level, is favorable for penetrating cell walls and cell membranes of bacteria and releasing the volatile oil into the bacterial cells to play a role, and thus the antibacterial activity of the volatile oil is enhanced. Meanwhile, the composite volatile oil ethosome provided by the invention has lower ethanol content and lower irritation.
The composite volatile oil ethosome prepared by the preparation method disclosed by the invention encapsulates the active ingredient composite volatile oil in the ethosome, and the composite volatile oil can be slowly released through ethosome coating, so that a more long-acting antibacterial effect is exerted.
The invention has the beneficial effects that:
the composite volatile oil ethosome provided by the invention has lower alcohol content, is a healthier food preservative and an antibacterial agent with lower irritation and higher efficiency, has better antibacterial performance, and has larger diameter data of antibacterial rings of escherichia coli and staphylococcus aureus, and the diameter data can reach 20mm under better conditions; and because the composite volatile oil ethosome has higher encapsulation rate, stable overall performance and long antibacterial release time, the experimental verification proves that the composite volatile oil ethosome has slow weight loss trend and lower proportion, and the weight loss proportion is basically lower than 60 percent or even lower than 40 percent in a period of 4 months.
The preparation method of the composite volatile oil ethosome provided by the invention has the advantages of green and environment-friendly raw materials, low price and easiness in obtaining, simple and controllable preparation process, and excellent performance of the prepared composite volatile oil ethosome, and has the condition of industrial scale popularization and use.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a Tyndall effect plot of the composite volatile oil ethosome prepared in example 3;
FIG. 2 is a diagram showing the bacteriostatic effect of the composite volatile oil ethosome prepared in example 3 on Escherichia coli;
FIG. 3 is a diagram showing the bacteriostatic effect of the composite volatile oil ethosome prepared in example 3 on Staphylococcus aureus;
FIG. 4 is a graph showing the bacteriostatic effect of control example 1 on E.coli;
FIG. 5 is a graph showing the bacteriostatic effect of control example 1 on Staphylococcus aureus.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The preparation method of the composite volatile oil ethosome provided by the invention comprises the following steps:
s1: weighing 3% of soybean lecithin, 1.5% of phytosterol, 0.4% of compound volatile oil prepared by compounding litsea cubeba oil and tsaoko oil according to the mass ratio of 7 to 3 and 20% of absolute ethyl alcohol according to the weight parts, placing the mixture in a conical flask with a plug, and stirring the mixture by a magnetic stirrer at the speed of 700rpm until the mixture is completely dissolved to obtain an alcohol phase;
s2: weighing 0.1 percent of sodium di (lauramide glutamine) lysine serving as a surfactant according to the weight percentage, putting the sodium di (lauramide glutamine) lysine into a beaker, and adding 75 percent of distilled water to completely dissolve the sodium di (lauramide glutamine) lysine to obtain a water phase;
s3: stirring at 700rpm under 35 ℃ water bath and sealing conditions, dripping the water phase obtained in the step S2 into the alcohol phase obtained in the step S1 at the speed of 1mL/min, and continuing stirring at 700rpm after the water phase is completely added;
s4: homogenizing at 10000rpm for 20min with a high speed homogenizer, and filtering with 0.22 μm microporous membrane to obtain the final product.
Example 2
In contrast to example 1, the surfactant weighed in S2 was 0.1% dipalmitoyl glutathione.
Example 3
Different from the embodiment 1, the composite volatile oil in the S1 comprises the following components in percentage by mass of 9; in addition, the surfactant weighed in the S2 is 0.1% of sodium di (lauramide glutamine) lysine and 0.05% of dipalmitoyl glutathione, and the amount of the added distilled water is 74.95%.
Comparative example 1
In contrast to example 1, tween-80 was weighed as the surfactant in S2, and the amount added was 0.1%.
Comparative example 2
Unlike example 1, the complex volatile oil was not added to S1, and the amount of distilled water added to S2 was 75.4%.
Comparative example 3
Unlike example 1, no surfactant was added in the step S2, and 75.1% of distilled water was directly used as an aqueous phase.
The products prepared in examples 1-3 and comparative examples 1-3 were tested for the relevant properties as follows:
1. and (3) stability testing:
taking a proper amount of products prepared in examples 1-3 and comparative examples 1-3, placing the products in penicillin bottles, and observing the products by laser beam irradiation under a black background, wherein the result of the Tyndall effect test of the composite volatile oil ethosome prepared in example 3 is shown in figure 1, and according to figure 1, the composite volatile oil ethosome prepared in the example is in a colloidal state and shows a strong Tyndall effect.
Examples 1-3 and comparative examples 1-3 the results of the tests are shown in table 1 below:
table 1 table of stability test results
As shown in table 1, the composite volatile oil ethosome provided by the invention has higher encapsulation efficiency, so that the appearance and the stability are better. Comparative example 1 the ethosome prepared by using the conventional surfactant Tween-80 has obviously poor stability than the ethosome prepared by using the conventional surfactant Tween-80; comparative example 2 does not add the core component of the composite volatile oil, so the stability is stronger, but the composite volatile oil does not belong to the ethosome of the composite volatile oil, and the corresponding fresh-keeping and bacteriostatic functions are poorer; comparative example 3 no surfactant was added and the appearance and stability of the resulting ethosome were the worst.
2. And (3) testing the release rate:
taking a proper amount of the products prepared in the examples 1-3 and the comparative examples 1 and 3, placing the products in a constant temperature and humidity box, setting the temperature at 25 ℃ and the humidity at 80%, and measuring the weight loss data of the prepared ethosome within the initial period, 1 month, 2 months, 3 months and 4 months respectively, wherein the test results are shown in the following table 2:
table 2 release rate test results table
| Item | 1 month | 2 months old | 3 months old | 4 months old |
| Example 1 | 20.1％ | 30.4％ | 38.4％ | 45.6％ |
| Example 2 | 22.3％ | 34.7％ | 43.3％ | 51.2％ |
| Example 3 | 15.6％ | 24.5％ | 32.3％ | 38.9％ |
| Comparative example 1 | 39.1％ | 56.0％ | 73.8％ | 83.2％ |
| Comparative example 2 | 1.2％ | 1.3％ | 2.1％ | 2.6％ |
| Comparative example 3 | 52.1％ | 68.3％ | 80.4％ | 91.2％ |
As can be seen from Table 3, the data of examples 1-3 show that the composite volatile oil ethosome has high encapsulation rate, obviously improved slow release performance, long release time, slow weight loss and low proportion, and the weight loss is lower than 60 percent or even lower than 40 percent in a 4-month period; experiments prove that the release rate of the composite volatile oil ethosome prepared under the optimal condition can be as long as one year. Meanwhile, the ethosome prepared in the comparative example 1 or the comparative example 3 has poor stability under the condition of adding a conventional surfactant or not adding the surfactant, and meanwhile, the release rate of the volatile oil is relatively high, the weight loss trend is obvious and the proportion is large, and the volatile oil exceeds 80 percent and even reaches 90 percent in a 4-month period; comparative example 2 does not contain the compound volatile oil, and the change of the mass is not large.
3. And (3) testing the antibacterial performance:
and (3) testing conditions: the environment was sterilized with an ultraviolet lamp before the test runs, all of which were performed alongside the alcohol burner flame.
And respectively picking the escherichia coli and staphylococcus aureus colonies which are activated and cultured in advance, adding the escherichia coli and staphylococcus aureus colonies into 1mL of sterile water, fully oscillating and diluting. 100 μ L of diluted cell suspension was taken by a pipette gun, added to a petri dish, immediately spread with an applicator, shaken with sterilized forceps on the flame of an alcohol lamp, then the sterilized filter paper pieces were held by forceps on a petri dish, 10 μ L each of the ethosome prepared in examples 1 to 3 and comparative examples 1 to 3 was dropped, a circular small filter paper piece with the ethosome attached was attached to a culture medium, gently pressed with forceps, and immediately closed with forceps, and the culture dish was placed in a constant temperature incubator at 37 ℃ for inverted culture for 48 hours.
The bacteriostatic effect test of the ethosome of the compound volatile oil prepared in the embodiment 3 is shown in fig. 2 and fig. 3, wherein fig. 2 is a bacteriostatic effect diagram of escherichia coli, and fig. 3 is a bacteriostatic effect diagram of staphylococcus aureus.
The test results of examples 1-3 and comparative examples 1-3 are shown in table 3 below:
table 3 bacteriostatic property test result table
According to the figure 2, the figure 3 and the table 3, the diameter data of the composite volatile oil ethosome aiming at escherichia coli and staphylococcus aureus are larger, and can reach 20mm or more under better conditions, which indicates that the composite volatile oil ethosome has high antibacterial capacity. Meanwhile, the size of the inhibition zone of the comparative examples 1-3 is obviously lower than that of the composite volatile oil ethosome prepared by the invention, which indicates that the inhibition capacity of the conventional ethosome is obviously lower than that of the composite volatile oil ethosome prepared by the invention.
Comparative example 1
Taking the following volatile oils: respectively testing the bacteriostatic effects of the single volatile oil on escherichia coli and staphylococcus aureus according to the bacteriostatic performance test method by using A-clove oil, B-tsaoko oil, C-anise oil, D-litsea cubeba oil, E-citronella oil and F-tangerine peel oil. The results are shown in table 4, fig. 4 and fig. 5, wherein fig. 4 is a bacteriostatic effect graph of escherichia coli, and fig. 5 is a bacteriostatic effect graph of staphylococcus aureus.
TABLE 4 bacteriostatic properties test results of single volatile oil
From the results, the bacteriostatic performance of the single volatile oil is integrally lower than that of the composite volatile oil ethosome provided by the invention, and as can be seen from the graphs in fig. 4 and 5, the single volatile oil has a direct bacteriostatic effect, and the volatile oil of the litsea cubeba and the volatile oil of the amomum tsao-ko also shows a gas-phase bacteriostatic effect, so that the growth of staphylococcus aureus and escherichia coli colonies is very slow and the colonies are less than other volatile oils in places where the volatile oil is not in direct contact. The volatile oil of the litsea cubeba has particularly remarkable gas phase bacteriostasis effect.
Note that, the technical features of the above embodiments may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the above embodiments are not described, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description in the present specification.
The above examples only express several embodiments of the present application, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the concept of the present application, and these are all within the scope of protection of the present application. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. The composite volatile oil ethosome is characterized by comprising the following raw materials in percentage by weight except water: 0.1-3% of compound volatile oil, 0.5-5% of phospholipid, 0.01-1% of membrane fluidity regulator, 10-20% of ethanol and 0.1-0.4% of surfactant.
2. The complex volatile oil ethosome of claim 1, wherein said complex volatile oil is selected from at least two of the following volatile oils: <xnotran> , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , D- , , , , , , , . </xnotran>
3. The composite volatile oil ethosome according to claim 1, wherein said phospholipid comprises a natural phospholipid and/or a synthetic lipid;
preferably, the natural phospholipid comprises one or more of soy phosphatidylcholine, egg lecithin, soy lecithin, lysolecithin, sphingomyelin, phosphatidyl, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, diphosphatidylglycerol, and plasmalogen.
4. The composite volatile oil ethosome of claim 1, wherein said membrane fluidity modifier comprises a phytosterol and/or cholesterol;
preferably, the phytosterols comprise one or more of 4-methyl free sterol, 4-methyl sterol, 4' -dimethyl sterol, BETA-sitosterol, stigmasterol, campesterol and brassicasterol.
5. The composite volatile oil ethosome according to claim 1, wherein said surfactant is a linear alkyl acyl peptide structure surfactant;
preferably, the surfactant comprises sodium di (lauramidoglutamine) lysine and/or dipalmitoyl glutathione.
6. A method of preparing a composite volatile oil ethosome according to any one of claims 1 to 5, comprising: and carrying out first mixing and homogenization on the raw materials and water to obtain the composite volatile oil ethosome.
7. The preparation method according to claim 6, wherein the first mixing is specifically: dissolving the surfactant in water to form an aqueous phase; dissolving said phospholipid, said membrane fluidity modifier, and said complex volatile oil in said ethanol to form an alcohol phase;
second mixing the aqueous phase and the alcohol phase.
8. The method of claim 7, wherein the second mixing comprises dropping the aqueous phase into the alcohol phase at a rate of 1 to 5mL/min under incubation and sealing conditions.
9. The production method according to claim 8, wherein the second mixing is performed in a stirred state, wherein a stirring rate at the time of the dropwise addition is 300 to 800rpm, and a stirring rate after the completion of the dropwise addition is 500 to 800rpm.
10. The method of any one of claims 6-8, wherein the homogenization is performed at 6000-10000 rpm for 5-10 min.
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| CN1957924A (en) * | 2006-11-22 | 2007-05-09 | 上海微纳科技有限公司 | Nano liposome of tea plant oil, and preparation method |
| CN108721604A (en) * | 2018-08-21 | 2018-11-02 | 山东大学 | A kind of -4 alcohol plastid of extrasin beta and its preparation process |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1957924A (en) * | 2006-11-22 | 2007-05-09 | 上海微纳科技有限公司 | Nano liposome of tea plant oil, and preparation method |
| CN108721604A (en) * | 2018-08-21 | 2018-11-02 | 山东大学 | A kind of -4 alcohol plastid of extrasin beta and its preparation process |
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