CN114592017B - Method for preparing essential oil emulsion by enzyme method and application of essential oil emulsion in bacteriostat - Google Patents
Method for preparing essential oil emulsion by enzyme method and application of essential oil emulsion in bacteriostat Download PDFInfo
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- A61K47/6951—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
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- A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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Abstract
The invention discloses a method for preparing essential oil emulsion by an enzyme method and application of the essential oil emulsion in a bacteriostatic agent, and belongs to the field of emulsion preparation. The method for preparing the essential oil emulsion by the enzymatic method comprises the following steps: (1) Adding enzyme into the starch water solution, performing enzymolysis, and cyclizing to obtain a cyclized solution; (2) Uniformly mixing the essential oil and the auxiliary emulsifier to obtain a mixed solution of the essential oil and the auxiliary emulsifier; (3) Adding a mixed solution of essential oil and an auxiliary emulsifier into the cyclization solution, and embedding to obtain an embedding solution; (4) Adding an emulsifying agent into the embedding solution in the step (3), emulsifying, shearing and homogenizing to obtain the essential oil emulsion. The emulsion prepared by the invention is extremely stable, and does not delaminate when being placed for 60 days at 20, 40 and 60 ℃, thereby being beneficial to the storage, transportation and use of products; the dilution stability is good, the stability can be kept after dilution in any proportion, and no layering phenomenon occurs.
Description
Technical Field
The invention relates to a method for preparing essential oil emulsion by an enzyme method and application of the essential oil emulsion in a bacteriostat, and belongs to the field of emulsion preparation.
Background
Plant essential oils (essential oils) are volatile oily compounds with strong odors extracted from various tissues of aromatic plants, and the main components are several tens or even more hundred kinds of alkaloids, flavonoids, phenolic acids, monoterpenes, isoflavones, aldehydes, oxygenated terpenes, non-oxygenated terpenes, etc., and most of them are simple lipophilic compounds, among which citral, cinnamaldehyde, carvacrol, thymol, eugenol, etc. are common. The plant essential oil is usually obtained by water distillation, steam and water distillation, solvent extraction or supercritical fluid extraction. Because of the increasing current demand for the development of natural, safe, effective healthy products, the search for plant essential oils has been one of the research hotspots.
The plant essential oil is generally liquid at normal temperature, and part of the single-component essential oil is crystalline powder at normal temperature, and is generally colorless or light yellow. The plant essential oil has a wide boiling point range, a specific gravity range of 0850-1.065 at 70-300 ℃, a pH value of usually acidic, a neutral optical rotation (+ 97-117 DEG) and a refractive index (143-161) of higher. Plant essential oil is extremely insoluble in water, is easily dissolved in organic solvents such as alcohols and ethers, is easily volatilized at normal temperature, is sensitive to light, heat and oxygen, and is easily deteriorated after long-term exposure to the environment.
Currently, relatively few reports are reported on improving the application of essential oils. Liu Tonghui A method for embedding essential oil (Liu Tonghui. One-step method for preparing cyclodextrin-plant essential oil compound and its antibacterial property research [ J ]. University of Jiangnan, 2021.) is disclosed, but the prepared microcapsule still has the problem of limited emulsion preparation, namely that the oil phase is not more than 20% and the water phase is not less than 60%.
Disclosure of Invention
[ technical problem ]
The plant essential oil is almost insoluble in water, is volatile and has poor stability, and has certain hydrophobicity. Therefore, the prepared water aqua has the problems of uneven dispersion and poor stability.
Technical scheme
In order to solve the problems, the invention combines emulsification and an enzymatic cyclodextrin preparation system to prepare the essential oil emulsion, thereby greatly improving the water solubility of the essential oil emulsion. Meanwhile, the cyclodextrin prepared by cyclizing the starch substrate through the enzymatic action is used for embedding the essential oil, so that the method has the advantages of improving the physicochemical property of the essential oil, preventing volatilization, improving the solubility, reducing the sensitivity to the environment and the like. In addition, starch is used as a raw material to prepare the emulsion, so that the use amount of cyclodextrin can be saved, the cost is greatly reduced, the process is simplified, the problem of insufficient water solubility of the cyclodextrin can be solved by introducing the emulsifier, and the use amount of the emulsifier is reduced when the cyclodextrin exists, so that the system safety is improved.
The first object of the invention is to provide a method for preparing an essential oil emulsion by an enzymatic method, which comprises the following steps:
(1) Adding enzyme into the starch water solution, performing enzymolysis, and cyclizing to obtain a cyclized solution;
(2) Uniformly mixing the essential oil and the auxiliary emulsifier to obtain a mixed solution of the essential oil and the auxiliary emulsifier;
(3) Adding the mixed solution of the essential oil and the auxiliary emulsifier in the step (2) into the cyclization solution in the step (1) to perform embedding to obtain an embedding solution;
(4) Adding an emulsifying agent into the embedding solution in the step (3), emulsifying, shearing and homogenizing to obtain the essential oil emulsion.
In one embodiment of the invention, the mass percentage of each substance in the step (1), the step (2) and the step (3) in the essential oil emulsion is 1.5-2% of starch, 1-40% of essential oil, 10-20% of emulsifying agent, 2.5-10% of auxiliary emulsifying agent and the balance of water, wherein the total amount is 100%.
In one embodiment of the present invention, the starch in step (1) comprises one or more of wheat starch, corn starch, tapioca starch, waxy corn starch; further preferred are corn starch and tapioca starch.
In one embodiment of the present invention, the starch aqueous solution in step (1) is obtained by dissolving starch in water, stirring at 200-500 rpm at 40-60 ℃ for 5-15 min, and mixing until the liquid is clear and transparent.
In one embodiment of the present invention, the enzyme of step (1) comprises one or more of α -cgtase, β -cgtase, γ -cgtase, wherein α -cgtase, β -cgtase, γ -cgtase are all commercially available; the enzyme activity of the alpha-CGTase is 200U/mL, the enzyme activity of the beta-CGTase is 30U/mL, and the enzyme activity of the gamma-CGTase is 6U/mL.
In one embodiment of the invention, the enzyme of step (1) is added in an amount of 1 to 8U/g starch.
In one embodiment of the present invention, the enzymolysis in the step (1) is performed by heating to 60-90 ℃ and stirring at 200-500 rpm for 5-15 min.
In one embodiment of the present invention, the cyclization in step (1) is carried out by cooling to 40-60℃and stirring at 200-500 rpm for 30-50 min.
In one embodiment of the present invention, the essential oil in step (2) includes one or more of cinnamaldehyde, carvacrol, thymol, and eugenol.
In one embodiment of the present invention, the co-emulsifier in step (2) comprises one or both of ethanol and acetic acid, and more preferably ethanol.
In one embodiment of the invention, the uniform mixing in the step (2) is preheating for 30-60 s at 40-60 ℃.
In one embodiment of the present invention, the embedding in step (3) is performed at 40-60℃and 200-500 rpm for 5-15 min.
In one embodiment of the present invention, the emulsifier in the step (4) includes one or more of polyoxyethylated castor oil 30, tween, span; tween includes tween 20, tween 40, tween 60, tween 80; span includes span 20, span 40, span 60, span 80, and more preferably polyoxyethylene castor oil 30 or tween 80.
In one embodiment of the invention, the emulsification in the step (4) is carried out at 40-60 ℃ and 200-500 rpm for 15-30 min; further preferably 50℃and 300 rpm.
In one embodiment of the present invention, the shearing and homogenizing in the step (4) is performed at 5000rpm to 15000rpm for 1 to 3 minutes, and the homogenizing is performed at 300 to 500Bar pressure by using a homogenizer.
The second purpose of the invention is the essential oil emulsion prepared by the method.
The third purpose of the invention is to apply the emulsion essential oil of the invention in the fields of medicine, daily chemicals and agriculture.
A fourth object of the present invention is to provide a bacteriostatic agent comprising the essential oil emulsion of the present invention.
[ advantageous effects ]
(1) The invention prepares cyclodextrin by using starch as a raw material through an enzymatic method and combines an emulsifying system to prepare an emulsion system of essential oil, so that the raw material is easy to obtain, the process is simple, the operation is convenient, and the industrial and large-scale production is very easy to realize.
(2) According to the invention, an enzymatic system is introduced into the traditional emulsification technology, the cyclodextrin property of 'inner thinning and outer thinning' is utilized to clathrate the essential oil, so that the stability of the essential oil is greatly improved, and a novel method is provided for enhancing the application performance of the essential oil.
(3) The emulsion prepared by the invention is extremely stable, and does not delaminate when being placed for 60 days at 20, 40 and 60 ℃, thereby being beneficial to storage, transportation and use of products.
(4) The essential oil emulsion prepared by the invention has good dilution stability, can be diluted in any proportion and is stable, no layering phenomenon occurs, and the conditions of poor stability and insufficient water solubility of the essential oil are obviously improved.
(5) The essential oil emulsion product prepared by the invention has a good antibacterial effect.
Drawings
FIG. 1 shows the results of stability tests for different emulsifiers in example 2.
FIG. 2 shows the results of the stability test of the various coemulsifiers of example 3.
FIG. 3 is a graph showing the results of the stability test for the amounts of absolute ethanol used as the various co-emulsifiers in example 4; wherein, (a) is 2.5g of absolute ethyl alcohol, and the mass ratio of the emulsifier polyoxyethylene castor oil 30 to the co-emulsifier absolute ethyl alcohol is 4:1, a step of; (b) 5g of absolute ethyl alcohol, wherein the mass ratio of the emulsifier polyoxyethylene castor oil 30 to the co-emulsifier absolute ethyl alcohol is 2:1, a step of; (c) 10g of absolute ethyl alcohol, wherein the mass ratio of the emulsifier polyoxyethylene castor oil 30 to the co-emulsifier absolute ethyl alcohol is 1:1, a step of; (d) 40g of absolute ethyl alcohol, wherein the mass ratio of the emulsifier polyoxyethylene castor oil 30 to the co-emulsifier absolute ethyl alcohol is 1:4, a step of; (e) 0g of absolute ethanol.
FIG. 4 is a graph showing the stability test results of various eucalyptol compounds of example 5; wherein, the addition amount of the eucalyptol in (a) to (f) is 1 g, 5g, 10g, 20g, 40g and 60g.
FIG. 5 is the test results of example 13; wherein, from top to bottom, the first row is cinnamaldehyde essential oil emulsion, the third row is thymol essential oil emulsion, the fifth row is carvacrol essential oil emulsion, the seventh row is eugenol essential oil emulsion, and the second, fourth, sixth and eighth rows are covers; the concentration in the first column was 2.5mg/ml, after which each column was 50% lower than the previous column.
Detailed Description
The following description of the preferred embodiments of the present invention is provided for better illustration of the invention, and should not be construed as limiting the invention.
The testing method comprises the following steps:
1. the stability of the emulsion was evaluated by direct observation:
the emulsion was placed in a centrifuge tube and allowed to stand at various temperatures (20, 40, 60 ℃) and the emulsion was observed with the naked eye periodically to see if it was uniform and stable.
2. Dilution performance evaluation of emulsion:
after the system is subjected to gradient dilution, the direct observation method is adopted to carry out: part of the liquid was sucked from the prepared system, subjected to two-fold gradient dilution, stored in a centrifuge tube, and left at room temperature, and the state of the diluted liquid was observed with naked eyes at regular intervals.
3. Evaluation of antibacterial performance of essential oil emulsion products:
mixing the solid culture medium with the double concentration which is not coagulated with the essential oil, pouring the essential oil into a flat plate, respectively inoculating 3 coliform bacteria liquid which is diluted by 100 times after 1 microliter of the solid culture medium is subjected to expansion culture for 8 hours on each culture medium, and observing colony growth conditions after the solid culture medium is subjected to constant temperature culture in a 37 ℃ incubator for 12 hours to obtain the minimum antibacterial concentration of a system to be tested.
Example 1
A method for preparing an essential oil emulsion by an enzyme method, which comprises the following steps:
(1) Adding 1.5g of corn starch into 66g of distilled water, and stirring for 10min at 50 ℃ and 300rpm to obtain a clear and transparent starch aqueous solution;
(2) Adding 4U/g (starch) beta-CGTase into starch water solution, heating to 90 ℃, stirring at 300rpm for 10min for enzymolysis, cooling to 50 ℃, stirring at 300rpm for 30min for cyclization, and obtaining cyclized solution;
(3) Preheating 20g of eucalyptol and 2.5g of absolute ethyl alcohol serving as a coemulsifier at 50 ℃ for 1 minute to obtain a mixed solution of essential oil and the coemulsifier;
(4) Adding the mixed solution of the essential oil and the auxiliary emulsifier in the step (3) into the cyclization solution in the step (2), and stirring at 50 ℃ and 300rpm for 30min to obtain an embedding solution;
(5) Adding 10g of emulsifier polyoxyethylene castor oil 30 (EL-30) into the embedding solution in the step (4), stirring at 50 ℃ and 300rpm for 15min for emulsification, shearing at 10000rpm for 2min, and homogenizing under 500Bar pressure to obtain the essential oil emulsion.
Example 2 selection of emulsifier
The emulsifier EL-30 in example 1 was adjusted to Tween 80, sodium lauroyl sarcosinate (LS-30N) and polyoxyethylene octyl phenol ether-10 (OP-10), and the other was kept the same as in example 1 to obtain an essential oil emulsion.
The resulting emulsion was tested as follows:
TABLE 1
As can be seen from table 1, fig. 1: when the El-30 is used as an emulsifier, the prepared eucalyptol emulsion system is stable, is uniform and stable and is not layered after being placed for 60 days at 20, 40 and 60 ℃, and can maintain a good uniform system after being diluted by water according to any proportion; the HLB value of Tween 80 is about 15, and the emulsion taking Tween 80 as the emulsifier is an ideal emulsifier, but the emulsion taking Tween 80 as the emulsifier can be diluted only at a multiple of 1-10, and further dilution can damage the water-soluble property of the emulsion, so that layering occurs, and the emulsion is not taken as an optimal scheme. When sodium lauroyl sarcosinate (LS-30N) and polyoxyethylene octylphenol ether-10 (OP-10) are used as emulsifiers, the resulting eucalyptol emulsion is unstable and delaminates rapidly, possibly due to the incompatibility of these emulsifiers with the system or the preparation method.
Example 3 selection of Co-emulsifier
The co-emulsifier absolute ethanol of example 1 was adjusted to acetic acid, and the other was kept the same as example 1 to obtain an essential oil emulsion.
The resulting emulsion was tested as follows:
TABLE 2
As can be seen from table 2, fig. 2: the system prepared by using acetic acid as the auxiliary emulsifier has weak layering phenomenon, and the system prepared by using ethanol as the auxiliary emulsifier is uniformly milky and has good stability, so that the ethanol is preferable as the auxiliary emulsifier.
Example 4 amount of co-emulsifier
The amounts of the auxiliary emulsifier absolute ethyl alcohol in the example 1 are adjusted to 0g, 2.5g, 5g, 10g and 40g, and the amounts of the water are adjusted to 68.5g, 66g, 63.5g, 58.5g and 28.5g, so that the total percentage is 100%, and the other parts are kept consistent with the example 1, so that the essential oil emulsion is obtained.
The resulting emulsion was tested as follows:
TABLE 3 Table 3
As can be seen from table 3, fig. 3: when the ratio of the emulsifier to the auxiliary emulsifier is higher than 1:1, the system is stable; as shown in fig. 3 (a), (b), and (c). When the proportion of ethanol is high, starch and decomposition products thereof have low solubility in ethanol, and precipitate is separated out, so that the system is obviously unstable and layered, as shown in (d) of fig. 3. At the same time, comparing the stable ethanol-containing system with (e) in FIG. 3, it was found that the whole of the system was stable when ethanol was not added to the embedding system, but there was a little white floccule at the junction of the surface and the cup wall, and it was suspected that the lack of ethanol had an influence on the balance of the system.
Considering that ethanol is inflammable and explosive liquid at normal temperature and normal pressure, reducing the content of ethanol in a system is beneficial to transportation and storage of pesticide products, and finally, the adding amount of the ethanol serving as an auxiliary emulsifier is 1/4 of that of the emulsifier, namely: the amount of the co-emulsifier ethanol is 2.5g.
EXAMPLE 5 Eucalyptus extract dosage
The addition amounts of eucalyptol in example 1 were adjusted to 1, 5, 10, 20, 40 and 60g, and the amounts of water were adjusted to 85, 81, 76, 66, 46 and 26g, so that the total percentage was 100%, and the other materials were kept the same as in example 1, to obtain an essential oil emulsion.
The resulting emulsion was tested as follows:
TABLE 4 Table 4
As can be seen from table 4, fig. 4: the dosage of eucalyptol reaches 40g at most, and when the dosage is further increased, the emulsifier can not balance water and oil phases, so that the phenomenon of instability occurs.
EXAMPLE 6 amount of emulsifier
The amounts of the emulsifiers EL-30 in example 1 were adjusted to 0.625, 1.25, 2.5, 5, 10, 15 and 20g, and the amounts of water were adjusted to 75.375, 74.75, 73.5, 72, 66, 61 and 56g, so that the total percentage was 100%, and the other components remained the same as in example 1, to obtain an essential oil emulsion.
The resulting emulsion was tested as follows:
TABLE 5
As can be seen from table 5: under the conditions of the current system proportion and the preparation process, the dosage of the emulsifier EL-30 is lower than 5g, namely the concentration is lower than 5%, and the system is easy to be unstable. Higher amounts of emulsifier increase the viscosity of the system and also increase the cost of preparation. Meanwhile, when the amount of the emulsifier is too high (20%), a significant delamination phenomenon may occur because it is more easily combined with eucalyptol. Therefore, 10g of the optimum amount of the emulsifier EL-30 is preferable.
EXAMPLE 7 time of emulsion reaction
The time of the emulsification reaction in example 1 was adjusted to 0, 10, 20, 30 minutes, and the other conditions were kept the same as in example 1, to obtain an essential oil emulsion.
The resulting emulsion was tested as follows:
TABLE 6
As can be seen from table 6: the emulsion layering is obvious due to the fact that the emulsion layering is too short, the content of eucalyptus essential oil is possibly high, the eucalyptus essential oil cannot be completely embedded by a single enzyme inclusion system, and an appropriate amount of emulsifying agent can be used for emulsifying the incompletely embedded essential oil to form a double-acting water-soluble system. Meanwhile, the time cost is considered, and the emulsification reaction time of 15min is most suitable.
Example 8 optimization of starch species
The corn starch in example 1 was adjusted to waxy corn starch, wheat starch, tapioca starch, and the other materials were the same as in example 1 to obtain an essential oil emulsion.
The resulting emulsion was tested as follows:
TABLE 7
As can be seen from table 7: obvious layering occurs at the bottom of the waxy corn starch group, the color of the wheat starch group is uneven up and down, the color of the corn starch and tapioca starch system is uniform and stable, but the tapioca starch has high branched chain content, and the prepared emulsion has high viscosity and is not suitable for preparing an emulsion system. Corn starch is finally selected as the reaction substrate.
Example 9 optimization of starch concentration
The corn starch dosage in example 1 was adjusted to 0.5, 1, 1.5, 2, 4g, while the water dosage was adjusted to 67, 66.5, 66, 65.5, 63.5g, ensuring 100% overall percentage, and the other materials remained the same as in example 1, to obtain an essential oil emulsion.
The resulting emulsion was tested as follows:
TABLE 8
As can be seen from table 8: the starch content is too low, the essential oil cannot be completely embedded, the floating oil appears and has obvious layering, the starch content is high, the system is thick although the system is uniform, and the content of starch enzymolysis products is high. Too high a starch substrate may retrograde starch during the reaction to produce insoluble precipitation. Resulting in a final preferred 1.5g starch solution as substrate for the enzymatic inclusion reaction.
Example 10 optimization of enzyme dosage
The amounts of beta-CGTase used in example 1 were adjusted to 1U/g, 2U/g and 8U/g, and the other amounts were kept the same as in example 1 to obtain an essential oil emulsion.
The resulting emulsion was tested as follows:
TABLE 9
As can be seen from table 9: the addition amount of the enzyme of 4U/g is most suitable, namely when the addition amount of the enzyme is small, the enzymolysis degree of the starch is insufficient, and the aging and precipitation can influence the product property; too high an enzyme addition affects the cost of preparation.
Example 11 optimization of cyclization time
The cyclization times of example 1 were adjusted to 10, 20, 30, 40, 50, 60min, and the other were kept the same as in example 1 to obtain an essential oil emulsion.
The resulting emulsion was tested as follows:
table 10
As can be seen from table 10: when the reaction time is less than 30min, the eucalyptol part is separated out without embedding due to insufficient cyclodextrin yield, so that the stability of a system is affected; excessive cyclization time results in increased byproducts, and precipitation occurs due to starch aging and other reactions, which is unfavorable for system formation. Considering in combination, 30min is the most suitable cyclization reaction time.
Example 12 System suitability comparison of different essential oils
The essential oils in step (2) of the adjustment example 1 are cinnamaldehyde, carvacrol, thymol and eugenol, which are common plant essential oils, and the other essential oils are consistent with the essential oils of the example 1, so as to obtain an essential oil emulsion.
The obtained emulsion was subjected to stability and dilution performance evaluation, and the test results are shown in table 11 below:
TABLE 11
As can be seen from table 11: the essential oil emulsion obtained after the essential oil type of the example 1 is changed is stable and not layered, and still not layered after being placed at 20, 40 and 60 ℃ for 60 days, has good stability and is beneficial to storage, transportation and use of products. Meanwhile, the plant essential oil is stable after being diluted in any proportion, and layering phenomenon is avoided, so that the system well solves the problem that the stability and water solubility of the plant essential oil are poor.
Example 13 Effect of enzymatic System on essential oil bacteriostasis
Preparing a blank control group by adjusting the dosage of eucalyptus essential oil in the step (3) in the embodiment 1 to be 0 g;
the antibacterial performance of the pure essential oils (cinnamaldehyde, carvacrol, thymol, eugenol), the blank group and the essential oil emulsion system prepared in example 12 were evaluated.
The test results are shown in table 12 below:
table 12
As can be seen from table 12: when the essential oil emulsion obtained after the essential oil types of the embodiment 1 are changed, the essential oil emulsion has different degrees of improvement on the antibacterial effect compared with the original essential oil, and probably because the emulsion system after inclusion enables the essential oil to be distributed more uniformly in the culture medium, the antibacterial effect is improved, so that the system can well solve the problem of poor stability and water solubility of the plant essential oil, and simultaneously provides a new thought for the application of the essential oil in antibacterial aspect.
Comparative example 1 Using beta-cyclodextrin alone
A method of preparing an essential oil emulsion comprising the steps of:
1.5g of beta-cyclodextrin is dissolved in water, 20g of eucalyptol is added, then a mixture of 10gEL and 2.5g of ethanol is added, and finally 66g of water is added to prepare a 100g system, so as to obtain an essential oil emulsion;
after 20 minutes the phenomenon was observed, finding that: a small amount of precipitate appeared at the bottom of the emulsion, and after standing for 1 day, 10 days, and 30 days, the sample was observed, and the precipitate was found to increase. From this analysis, the beta-cyclodextrin system can indeed play a certain embedding effect, but due to the low solubility of cyclodextrin, the addition of the emulsifier and the co-emulsifier ethanol can further reduce the solubility of the embedded product, and finally the sample is precipitated, which is unfavorable for the stable preservation of the product.
Comparative example 2 Using gamma-cyclodextrin alone
A method of preparing an essential oil emulsion comprising the steps of:
1.5g of gamma-cyclodextrin is dissolved in water, 20g of eucalyptol is added, then a mixture of 10gEL and 2.5g of ethanol is added, and finally 66g of water is added to prepare a 100g system, so as to obtain an essential oil emulsion;
after 20 minutes the phenomenon was observed, finding that: the sample is observed after the emulsion is placed for 1 day, 10 days and 30 days, and the emulsion is found to be layered, wherein the upper layer is an oil layer, and the lower layer is a darker liquid. The analysis is as follows: the gamma-cyclodextrin may not be precisely matched with the eucalyptol due to the large pore size, and part of the eucalyptol does not enter the embedding system. At the same time, the successfully embedded eucalyptol sinks to generate liquid delamination due to the increase of density.
Meanwhile, because the production cost of the gamma-cyclodextrin is too high, the gamma-cyclodextrin is not generally used for embedding in the process flow.
Comparative example 3 gamma-and beta-cyclodextrin was used
A method of preparing an essential oil emulsion comprising the steps of:
1.5g of cyclodextrin (gamma-cyclodextrin and beta-cyclodextrin are 1:1) is dissolved in water, 20g of eucalyptol is added, then a mixture of 10gEL and 2.5g of ethanol is added, and finally water is added to prepare a 100g system, so as to obtain essential oil emulsion;
after 20 minutes the phenomenon was observed, finding that: the system is stable and uniform. After 1, 10 and 30 days of standing, the samples were observed and the system was found to remain uncollimated. Comparing the dilution stability, the samples were found to be stable at 10-fold dilution, but delamination occurred after 10 days of storage after 100-fold dilution. Moreover, compared with example 1, the preparation process of pure cyclodextrin is complex, the processing cost is high, and the economic cost is considered in the mass production of the process. In combination, the enzymatic method of example 1 is selected to be suitable.
Comparative example 4 sequential preparation System of starch solution- & gt eucalyptol- & gt mixture of emulsifier and coemulsifier
A method for preparing an essential oil emulsion by an enzyme method, which comprises the following steps:
(1) Adding 1.5g of corn starch into 66g of distilled water, and stirring for 10min at 50 ℃ and 300rpm to obtain a clear and transparent starch aqueous solution;
(2) Adding 4U/g beta-CGTase into starch water solution, heating to 90 ℃, stirring at 300rpm for 10min for enzymolysis, cooling to 50 ℃, stirring at 300rpm for 30min for cyclization, and obtaining cyclized solution;
(3) 10g of emulsifier polyoxyethylene castor oil 30 (EL-30) and 2.5g of co-emulsifier absolute ethanol are preheated for 1 minute at 50 ℃ to obtain a mixed solution of the emulsifier and the co-emulsifier;
(4) Adding 20g of eucalyptol into the cyclized solution in the step (2), stirring at 50 ℃ and 300rpm for 30min, and embedding to obtain an embedding solution;
(5) Adding the mixed solution of the emulsifier and the auxiliary emulsifier in the step (3) into the embedding solution in the step (4), stirring at 50 ℃ and 300rpm for 15min for emulsification, shearing at 10000rpm for 2min, and homogenizing under 500Bar pressure to obtain the essential oil emulsion.
The result shows that: the emulsion appeared to be significantly oily, and after 1, 10, 30 days of standing the sample was observed to find that the emulsion appeared to be significantly layered. From this analysis, the lack of the auxiliary emulsifier in the embedding process can reduce the combination effect of the eucalyptol and the enzymatic system, so that the eucalyptol is affected by the surface tension in the process of entering the water-soluble system and cannot well enter the pores of the cyclodextrin, and the final emulsion layering is obvious. Thus, the system was prepared by selecting the order of the starch solution→eucalyptol and coemulsifier→emulsifier mixture in example 1.
Comparative example 5
The shear homogenizing step in step (5) of example 1 was omitted, and the essential oil emulsion was obtained in the same manner as in example 1.
The result shows that: the surface of the emulsion has tiny oil drops. After 1, 10 and 30 days of standing, the samples were observed and the emulsion was found to delaminate. Therefore, the shearing homogenization can well reduce the size of the emulsion, so that the emulsion is better dispersed in the system, and the emulsion plays an important role in improving the storage of products.
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (5)
1. The method for preparing the essential oil emulsion by using the enzymatic method is characterized by comprising the following steps of:
(1) Adding enzyme into the starch water solution, performing enzymolysis, and cyclizing to obtain a cyclized solution;
(2) Uniformly mixing the essential oil and the auxiliary emulsifier to obtain a mixed solution of the essential oil and the auxiliary emulsifier;
(3) Adding the mixed solution of the essential oil and the auxiliary emulsifier in the step (2) into the cyclization solution in the step (1) to perform embedding to obtain an embedding solution;
(4) Adding an emulsifying agent into the embedding solution in the step (3), emulsifying, shearing and homogenizing to obtain the essential oil emulsion;
wherein, in the essential oil emulsion, the mass percentages of all substances in the step (1), the step (2) and the step (3) are 1.5-2% of starch, 1-40% of essential oil, 10-20% of emulsifying agent, 2.5-10% of auxiliary emulsifying agent and the balance of water, and the total is 100%;
the starch in the step (1) is corn starch; cyclization is carried out by cooling to 40-60 ℃ and stirring at 200-500 rpm for 30-50 min; the enzyme in the step (1) is beta-CGTase;
the auxiliary emulsifier in the step (2) is absolute ethyl alcohol;
the emulsification in the step (4) is carried out at 40-60 ℃ and 200-500 rpm for 15-30 min; the emulsifier is polyoxyethylene castor oil 30.
2. The method of claim 1, wherein the enzyme in step (1) is added in an amount of 1-8 u/g starch; the enzymolysis is carried out by heating to 60-90 ℃ and stirring at 200-500 rpm for 5-15 min.
3. The method of claim 1, wherein the essential oil of step (2) is one of cinnamaldehyde, carvacrol, thymol, eugenol.
4. An essential oil emulsion prepared by the method of any one of claims 1-3.
5. A bacteriostatic agent comprising the essential oil emulsion of claim 4.
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