CN115404128B

CN115404128B - Multifunctional essential oil composition and preparation method and application thereof

Info

Publication number: CN115404128B
Application number: CN202211359010.5A
Authority: CN
Inventors: 陈国生; 何永玲; 邓悦新
Original assignee: Guangdong Shunde Aromatic Shijia Natural Products Manufacturing Co ltd
Current assignee: Guangdong Shunde Aromatic Shijia Natural Products Manufacturing Co ltd
Priority date: 2022-11-02
Filing date: 2022-11-02
Publication date: 2023-01-24
Anticipated expiration: 2042-11-02
Also published as: CN115404128A

Abstract

The invention provides a multifunctional essential oil composition and a preparation method and application thereof, belonging to the technical field of plant essential oil. The preparation method comprises the following steps of drying mint, eucalyptus, lavender, clove, melissa officinalis, rosemary and patchouli by microwave, mixing and crushing, performing ultrasonic-assisted extraction by using a mixed solvent, filtering and concentrating to obtain extracted oil, performing enzymolysis on solids by using complex enzyme and fermentation by using a complex microbial inoculum, extracting a fermentation product by using a mixed solution of ionic liquid and acetonitrile, performing extraction by using a nano magnetic fluid to obtain extracted slurry, performing reverse extraction, mixing the obtained extract with the extracted oil, purifying by using a ceramic membrane, adding a polydopamine-modified porous silica microsphere for adsorption to obtain the multifunctional essential oil composition.

Description

Multifunctional essential oil composition and preparation method and application thereof

Technical Field

The invention relates to the technical field of plant essential oil, and particularly relates to a multifunctional essential oil composition and a preparation method and application thereof.

Background

The normal sleep plays an important role in learning and memory formation, and the reduction of long-term sleep time and the reduction of sleep quality are considered as sleep deprivation, which can cause a series of functional changes of emotion, learning and memory, immunity and the like, and particularly cause obvious damage to cognitive functions, such as learning and memory capacity reduction, reaction delay, attention dispersion, orientation, space disorder and the like. The prevention and treatment of the alertness decline caused by sleep deprivation in the long-term operation process by drug intervention has become a preferred research target of people, wherein the study and application of psychostimulants such as amphetamine and caffeine are the most extensive. Studies have shown that these drugs have a significant effect on combating the adverse effects of long-term continuous operations. Modafinil was first used by france for the treatment of narcolepsy, narcolepsy and spontaneous hypersomnia. Modafinil reduced subjective drowsiness and fatigue in sleep deprivation conditions and maintained better alertness, attention, cognition and psychomotor ability in the subject; however, studies have shown that modafinil can cause elevated blood pressure, increased heart rate, fatigue, decreased appetite, and even impaired memory.

Aromatherapy, as a non-pharmaceutical intervention means, combines the dual functions of art and treatment, comprehensively considers the needs of human physiology, intelligence and deep soul, and is a method which naturally resembles integral treatment. The aromatherapy mainly adopts essential oil extracted from plants as a medium, is developed into various dosage forms, and achieves a natural therapy of relieving mental stress, treating diseases and promoting human health through ways of inhalation, bathing, incense, massage, external application and the like. The aromatherapy substances or essential oils have been reported to have a wake-promoting effect. From some traditional Chinese medicines recorded as early as Shen nong Ben Cao Jing to the later aromatic resuscitation inducing traditional Chinese medicines, essential oil contained in the traditional Chinese medicines has the characteristics of good curative effect, small toxic and side effects and the like, and is widely used for refreshing and restoring consciousness and prolonging daily waking time. In addition, modern studies have shown that the refreshing effect of essential oils is related to neurotransmitter signals in different areas of the brain, possibly involving a variety of neurotransmitters such as monoamines including Norepinephrine (NA), 5-hydroxytryptamine (5-HT); the amino acids include gamma-aminobutyric acid (GABA), acetylcholine (Ach), glutamic acid (GLU), and the like.

Plant essential oil is called essential oil in the field of botany, volatile oil in the field of chemistry and medicine, aromatic oil in the field of commerce, and is a volatile oily liquid which exists in leaves, roots, peels, flowers and fruits of aromatic plants, can be distilled along with water vapor, has certain smell (generally fragrance or pungent taste), is colorless in color, belongs to a secondary metabolite of plants and enjoys the reputation of 'liquid gold'.

The current methods for extracting plant essential oil are quite a lot, such as steam distillation method, organic solvent extraction method, molecular distillation methodSupercritical CO ₂ The method comprises a fluid extraction method, a subcritical water extraction method, a microcapsule-double aqueous phase extraction method, an ultrasonic microwave assisted extraction method and the like, wherein the steam distillation method is widely applied due to the advantages of simple equipment, simple and convenient operation, low extraction cost and the like, and is also the most common method for extracting the plant essential oil at present. The plant essential oil contains various groups of active compounds, has various functional properties such as oxidation resistance, bacteriostasis, cancer resistance, tumor resistance, fresh keeping, mosquito repelling, insect prevention and the like, and is widely applied to cosmetics, washing products, food (mainly fruits, vegetables, meat and aquatic products), tobacco and wine, agriculture and other industrial essences all the time. The steam distillation method has the advantages of simple equipment, easy operation, environmental protection and the like, but the HD method also has a plurality of defects. In the method, because the plant raw material tissues are placed at a high temperature for a long time, heat-sensitive components contained in the plant raw material tissues are thermally decomposed, and easily hydrolyzed components are hydrolyzed and the raw materials are coked, so that the quality of essential oil products is adversely affected, and the defects of high energy consumption, long time consumption and the like exist. The organic solvent extraction method has the advantages of simple equipment, low investment and high extraction rate of essential oil, but a large amount of organic solvent is needed in the extraction process, the environment is seriously polluted, and the residual extraction solvent in the final essential oil product is difficult to remove. The traditional methods for extracting plant essential oil include some traditional extraction methods such as absorption method, squeezing method, adsorption method, etc. besides the steam distillation method and organic solvent extraction method described above, wherein the absorption method and adsorption method are suitable for extracting heat-sensitive valuable essential oil, and the process is complicated and takes long time; the squeezing method is the most traditional essential oil extraction method, and the essential oil product obtained by the method is impure, the yield is low, and the storage time of the finished product is short.

Disclosure of Invention

The invention aims to provide a multifunctional essential oil composition, a preparation method and an application thereof, and the multifunctional essential oil composition has the effects of refreshing, restoring consciousness, inducing resuscitation, relieving depression, relieving viscera, smoothing breath, relaxing people, enjoying mood, preventing cold, smoothing nasal obstruction, relieving dizziness and headache, stopping dizziness and carsickness, preventing mosquito bites and bites, reducing swelling and relieving itching, promoting blood circulation to remove blood stasis, dredging channels and collaterals, effectively treating traumatic injuries and sprains and the like.

The technical scheme of the invention is realized as follows:

the invention provides a preparation method of a multifunctional essential oil composition, which comprises the steps of drying mint, eucalyptus, lavender, clove, melissa officinalis, rosemary and patchouli by microwave, mixing and crushing to obtain plant material powder, performing ultrasonic-assisted extraction on the plant material powder by using a mixed solvent, filtering, concentrating filtrate to obtain extracted oil, performing enzymolysis on solid by using complex enzyme and fermentation by using a complex microbial inoculum to obtain a fermentation product, extracting the fermentation product by using a mixed solution of ionic liquid and acetonitrile, adding nano-magnetic fluid for extraction to obtain extracted slurry, adding the extracted slurry into an organic solvent for reverse extraction to obtain an extract, mixing the obtained extract with the extracted oil, purifying by using a ceramic membrane, concentrating, adding porous silica microspheres modified by polydopamine for adsorption to obtain multifunctional essential oil microspheres, namely the multifunctional essential oil composition.

As a further improvement of the invention, the method comprises the following steps:

s1, pretreatment of plant materials: microwave drying herba Menthae, eucalyptus globulus Labill, lavender, flos Caryophylli, herba Melissae axillaris, herba Rosmarini officinalis, and herba Agastaches, mixing, and pulverizing to obtain plant material powder;

s2, mixed solvent extraction: adding the plant material powder prepared in the step S1 into a mixed solvent, performing ultrasonic-assisted extraction, filtering, keeping solid, and removing the solvent from the filtrate under reduced pressure to obtain extracted oil;

the mixed solvent is a mixed solvent of water, ethanol, acetonitrile and petroleum ether;

s3, enzymolysis: adding the solid obtained in the step S2 into water, adding complex enzyme for enzymolysis, and inactivating enzyme to obtain an enzymolysis product;

s4, fermentation: adding activated saccharomyces cerevisiae, bacillus subtilis and lactobacillus plantarum into the enzymolysis product obtained in the step S3, fermenting, culturing, sterilizing and filtering to obtain a fermentation product;

s5, extracting ionic liquid: adding the fermentation product obtained in the step S4 into the mixed solution of ionic liquid and acetonitrile, and slowly introducing CO ₂ Oscillating, stopping introducing CO ₂ To obtain high-extraction ionic liquid-acetonitrile mixed solutionStanding and layering the solution, separating the solution, removing a water layer, adding nano magnetic fluid, performing oscillation adsorption, performing magnetic separation, washing, drying and filtering to obtain extraction slurry;

s6, reverse extraction of active substances: adding the extraction slurry prepared in the step S5 into an organic solvent, stirring and extracting, performing magnetic separation, and removing the organic solvent to obtain an extraction liquid;

s7, purification: uniformly mixing the extracted oil obtained in the step S2 and the extract liquid obtained in the step S6, filtering by adopting a small-aperture ceramic membrane, collecting filtrate obtained after small molecule filtration, and concentrating by adopting an ultrafiltration membrane to obtain purified volatile oil liquid;

s8, preparing porous silicon dioxide microspheres: dissolving alkyl orthosilicate in an organic solvent to obtain an oil phase; dissolving a pore-foaming agent and a surfactant in water to obtain a water phase; adding the oil phase into the water phase, emulsifying by an SPG rapid membrane, adjusting the pH value, heating for reaction and solidification, centrifuging, washing and drying to obtain the porous silicon dioxide microspheres;

s9, preparation of the modified porous silicon dioxide microspheres: uniformly dispersing the porous silica microspheres prepared in the step S8 in water, adding dopamine hydrochloride and a catalyst, heating for reaction, centrifuging, washing and drying to obtain modified porous silica microspheres;

s10, preparing a multifunctional essential oil composition: and (4) adding the modified porous silica microspheres obtained in the step (S9) into the purified volatile oil liquid obtained in the step (S7), stirring, adsorbing and filtering to obtain multifunctional essential oil microspheres, namely the multifunctional essential oil composition.

As a further improvement of the invention, the mass ratio of the mint, the eucalyptus, the lavender, the clove, the melissa officinalis, the rosemary and the patchouli in the step S1 is 1-3:1-2:3-5:2-3:1-2:3-5:5-7; the microwave drying power is 1000-1500W, and the time is 1-2h; the volume ratio of water, ethanol, acetonitrile and petroleum ether in the mixed solvent in the step S2 is 3-5:5-7:1-3:3-5; the solid-liquid ratio of the plant material powder to the mixed solvent is 1:3-5g/mL; the power of the ultrasonic auxiliary extraction is 500-700W, and the time is 30-50min.

As a further improvement of the invention, the mass ratio of the solid to the compound enzyme in the step S3 is 10; the complex enzyme is selected from at least two of cellulase, pectinase, hemicellulase, amylase and ligninase; the temperature adopted by the enzymolysis is 40-60 ℃, and the time is 2-4h; preferably, the complex enzyme is a mixture of cellulase and pectinase, and the mass ratio of the complex enzyme to the pectinase is 3-5:1.

As a further improvement of the invention, the inoculation amounts of the saccharomyces cerevisiae, the bacillus subtilis and the lactobacillus plantarum after the activation in the step S4 are respectively 3-5%, 2-4% and 0.5-1.5%; the fermentation culture condition is 37-40 ℃, and the time is 24-48h; the activation method comprises respectively inoculating Saccharomyces cerevisiae, bacillus subtilis and Lactobacillus plantarum in Gao's medium, culturing at 37-39 deg.C and 50-70r/min for 12-18h to obtain extract with bacteria content of 10 ⁸ -10 ⁹ cfu/mL strain seed solution; the mixed solution of the ionic liquid and the acetonitrile in the step S5 is a mixed solution of the ionic liquid and the acetonitrile according to the mass ratio of 5-7:1-2; the ionic liquid is at least one selected from 1,3-dimethylimidazole bistrifluoromethylsulfonyl imide salt, 1-ethyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-propyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-butyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-pentyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-hexyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-octyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-decyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-dodecyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-tetradecyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-hexadecyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-ethyl-1-methylpyrrolidine bistrifluoromethylsulfonyl imide salt, 1-propyl-1-methylpyrrolidine bistrifluoromethylsulfonyl imide salt, 1-butyl-1-methylpyrrolidine bistrifluoromethylsulfonyl imide salt, 1-butyltributyl-propylbistrifluoromethylsulfonyl imide salt, 1-butylphosphonium sulfonyl imide salt, tributyl-butylbistrifluoromethylsulfonyl imide salt, 1-butylbis-butylphosphonium salt, and tributyl-butylbistrifluoromethanesulfonylimide salt; said slow introduction of CO ₂ Middle CO ₂ At an introduction rate of 0.5 to 1mL/min, the introduction time is 30-50min; the nano magnetic fluid is 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid; the mass ratio of the fermentation product to the nano magnetic fluid to the mixed solution of ionic liquid and acetonitrile is 10-3.

As a further improvement of the invention, the mass ratio of the extraction slurry to the organic solvent in the step S6 is 2:3-5; the organic solvent is at least one selected from dichloromethane, trichloromethane, carbon tetrachloride, ethyl acetate, petroleum ether, acetonitrile, methyl acetate and n-hexane; in the step S7, the aperture of the small-aperture ceramic membrane is 2000-3000D, the aperture of the ultrafiltration membrane is 30-100nm, and the relative density of the purified volatile oil liquid is 1.4-1.7; the mass ratio of the extraction oil to the extraction liquid is 3-5:2-7.

As a further improvement of the present invention, the alkyl orthosilicate in step S8 is methyl orthosilicate or ethyl orthosilicate; the organic solvent is at least one selected from dichloromethane, trichloromethane, carbon tetrachloride, ethyl acetate, petroleum ether, acetonitrile, methyl acetate and n-hexane; the pore-foaming agent is selected from at least one of polyoxyethylene sorbitan fatty acid ester, polyethylene glycol octyl phenyl ether, cetyl trimethyl ammonium bromide, oxyethylene-oxypropylene triblock copolymer PEO20-PPO70-PEO20 and PEO106-PPO70-PEO 10; the surfactant is at least one of tween-20, tween-40, tween-60, tween-80, sodium dodecyl benzene sulfonate, sodium dodecyl sulfate, sodium dodecyl sulfonate, sodium tetradecyl benzene sulfonate, sodium tetradecyl sulfonate, sodium hexadecyl benzene sulfonate, and sodium octadecyl benzene sulfonate; the mass ratio of the alkyl orthosilicate, the pore-forming agent and the surfactant is 10-3; the pore diameter of the SPG rapid membrane is 5000-7000nm; the pH value is adjusted to 8-9, the temperature for heating, reacting and curing is 50-70 ℃, and the time is 3-5h; in the step S9, the mass ratio of the porous silicon dioxide microspheres to the dopamine hydrochloride to the catalyst is 20-32; the catalyst is Co containing 3-5wt% ²⁺ The temperature of the heating reaction is 45-55 ℃ and the time is 2-4h; the modification in step S10 is moreThe mass ratio of the porous silicon dioxide microspheres to the purified volatile oil liquid is 10-15.

As a further improvement of the invention, the method specifically comprises the following steps:

s1, pretreatment of plant materials: drying 1-3 parts by weight of mint, 1-2 parts by weight of eucalyptus, 3-5 parts by weight of lavender, 2-3 parts by weight of clove, 1-2 parts by weight of melissa officinalis, 3-5 parts by weight of rosemary and 5-7 parts by weight of patchouli under 1000-1500W microwave for 1-2h, mixing, crushing, and sieving with a 60-100 mesh sieve to obtain plant material powder;

s2, extracting by using a mixed solvent: adding the plant material powder prepared in the step S1 into a mixed solvent, wherein the solid-liquid ratio of the plant material powder to the mixed solvent is 1:3-5g/mL, performing 500-700W ultrasonic-assisted extraction for 30-50min, filtering, retaining solid, and removing the solvent from the filtrate under reduced pressure to obtain extracted oil;

the mixed solvent is a mixed solvent of water, ethanol, acetonitrile and petroleum ether, and the volume ratio of the water, the ethanol, the acetonitrile and the petroleum ether is 3-5:5-7:1-3:3-5;

s3, enzymolysis: adding 10 parts by weight of the solid obtained in the step S2 into 100 parts by weight of water, adding 1-2 parts by weight of complex enzyme for enzymolysis, and inactivating the enzyme at 100-105 ℃ for 15-20min to obtain an enzymolysis product;

the compound enzyme is a mixture of cellulase and pectinase, and the mass ratio of the compound enzyme to the pectinase is 3-5:1;

s4, fermentation: adding activated saccharomyces cerevisiae, bacillus subtilis and lactobacillus plantarum into the enzymolysis product obtained in the step S3, wherein the inoculation amounts are 3-5%, 2-4% and 0.5-1.5%, respectively, fermenting and culturing for 24-48h at 37-40 ℃, sterilizing by ultraviolet rays, and filtering to obtain a fermentation product;

the activation method comprises respectively inoculating Saccharomyces cerevisiae, bacillus subtilis and Lactobacillus plantarum in Gao's medium, culturing at 37-39 deg.C and 50-70r/min for 12-18h to obtain extract with bacteria content of 10 ⁸ -10 ⁹ cfu/mL strain seed solution;

s5, extracting ionic liquid: adding 10 parts by weight of the fermentation product obtained in the step S4 into 15-20 parts by weight of the ionic liquid-acetonitrile mixed solution, and introducing CO ₂ The feeding rate is 0.5-1mL/min, and the feeding time isOscillating for 30-50min, stopping introducing CO ₂ To obtain a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, adding 2-3 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating adsorption, magnetic separation, washing, drying and filtering to obtain extraction slurry;

the mixed solution of the ionic liquid and the acetonitrile is a mixed solution of the ionic liquid and the acetonitrile according to the mass ratio of 5-7:1-2;

s6, reverse extraction of active substances: adding 2 parts by weight of the extraction slurry prepared in the step S5 into 3-5 parts by weight of an organic solvent, stirring and extracting, carrying out magnetic separation, and removing the organic solvent to obtain an extraction liquid;

s7, purification: 3-5 parts by weight of the extracted oil obtained in the step S2 and 2-7 parts by weight of the extract obtained in the step S6 are uniformly mixed, the mixture is filtered by adopting a small-aperture ceramic membrane with the aperture of 2000-3000D, the filtrate obtained after small molecule filtration is collected, and the filtrate is concentrated by adopting an ultrafiltration membrane with the aperture of 30-100nm to obtain purified volatile oil liquid with the relative density of 1.4-1.7;

s8, preparing porous silicon dioxide microspheres: dissolving 10 parts by weight of methyl orthosilicate or ethyl orthosilicate in 50 parts by weight of organic solvent to obtain an oil phase; dissolving 2-3 parts by weight of pore-foaming agent and 0.5-1 part by weight of surfactant in 100 parts by weight of water to obtain a water phase; adding the oil phase into the water phase, emulsifying by using an SPG rapid membrane with the aperture of 5000-7000nm, adjusting the pH value to 8-9, heating to 50-70 ℃, reacting and curing for 3-5h, centrifuging, washing and drying to obtain the porous silica microspheres;

preferably, the pore-foaming agent is a mixture of a macroporous pore-foaming agent polyoxyethylene sorbitan fatty acid ester and a mesoporous pore-foaming agent PEO106-PPO70-PEO10, and the mass ratio is 5-7:3.

Under the action of the macroporous pore-foaming agent and the mesoporous pore-foaming agent, the silicon dioxide microspheres can form complex and compact pore channels, so that the plant essential oil can be fully adsorbed and slowly released in the later period.

S9, preparation of the modified porous silicon dioxide microspheres: uniformly dispersing 20 parts by weight of the porous silica microspheres prepared in the step S8 in 100 parts by weight of water, adding 25-32 parts by weight of dopamine hydrochloride and 2-3 parts by weight of catalyst, heating to 45-55 ℃, reacting for 2-4h, centrifuging, washing, and drying to obtain modified porous silica microspheres;

the catalyst is Co containing 3-5wt% ²⁺ The pH value of the Tris-HCl solution is 5-6;

s10, preparing the multifunctional essential oil composition: and (2) adding 10-15 parts by weight of the modified porous silica microspheres obtained in the step (S9) into 5-7 parts by weight of the purified volatile oil liquid obtained in the step (S7), stirring, adsorbing and filtering to obtain multifunctional essential oil microspheres, namely the multifunctional essential oil composition.

The invention further protects the multifunctional essential oil composition prepared by the preparation method.

The invention further protects the application of the multifunctional essential oil composition in preparing related products for refreshing, restoring consciousness, inducing resuscitation, resolving stagnation, preventing cold, smoothing nasal obstruction, relieving dizziness and headache, stopping dizziness and car sickness, preventing mosquito bites and detumescence and relieving itching after biting.

The invention has the following beneficial effects:

according to the invention, all the components are firstly dried by microwaves, compared with the traditional drying method, for example, the drying method can expose plant materials in high-temperature air for a long time to cause terpenes to be degraded into monoterpenes, and simultaneously alcohols are esterified to form esters, so that the composition of part of essential oil in the later period is changed. Vacuum drying significantly changes the type and amount of active ingredients of the essential oil due to long vacuum treatment. Infrared drying causes oxidation and chemical structural rearrangement, which results in the production of new volatile compounds that differ significantly from the structure of the plant material itself. The microwave drying method adopted in the invention can improve the internal temperature of the plant material to be higher, cause the damage of plant cells and the rupture of cell walls, and is beneficial to the release of volatile compounds.

The invention utilizes the mixed solvent of water, ethanol, acetonitrile and petroleum ether to extract under the action of ultrasonic wave, the four-phase system of the water-ethanol-acetonitrile-petroleum ether mixed solvent comprises polar solvents of water, ethanol, acetonitrile and the like, and simultaneously comprises nonpolar solvents of petroleum ether, and simultaneously, after microwave drying, the cells of the plant material powder are cracked and damaged, and are further extracted under the assistance of ultrasonic wave to be further cracked, thereby being beneficial to the dissolution of active substances and the similar dissolution of the active substances by the solvent, and being dissolved in the solvent, thereby improving the solubility of the active substances in the solvent and improving the extraction rate.

The solid obtained by further filtering is subjected to enzymolysis by adopting a complex enzyme which is a mixture of cellulase and pectinase, so that the cell walls, pectin and the like of the plant cells are subjected to enzymolysis, a large amount of active substances in the cells are further dissolved out, and the extraction rate is improved; the fermentation technology is adopted to further destroy plant cells, promote the dissolution of active substances and further improve the extraction rate, and meanwhile, the fermentation process is also beneficial to generating a plurality of active aromatic small molecules which are also beneficial to improving the effects of preventing cold, smoothing nasal obstruction, relieving dizziness and headache, stopping dizziness and carsickness, preventing mosquito bites and swelling and relieving itching after biting, and the like.

The enzymolysis product is further extracted by the mixed solution of ionic liquid and acetonitrile, the enzymolysis product contains a large amount of polysaccharide, protein, peptide, oligosaccharide and other impurities, the ionic liquid is extracted by the mixed solution of ionic liquid and acetonitrile, the ionic liquid consists of positively charged ions and negatively charged ions, the ionic liquid is in a liquid state at the temperature of between 100 ℃ below zero and 200 ℃, the ionic liquid can not be steam to generate harmful gas causing air pollution generally, and the ionic liquid can be repeatedly used for a plurality of times. Hydrophobic ionic liquid is not soluble with water, substances dissolved in water can be extracted by utilizing the principle of 'similarity and compatibility', and an ionic liquid-acetonitrile mixed solvent is used as an extracting agent for extraction and separation, so that the problems of high viscosity and high cost of the ionic liquid can be solved, and the extraction of active substances can be promoted, thereby achieving good effects of separating impurities and extracting the active substances; meanwhile, in the extraction process, CO is slowly introduced ₂ The polarity of the mixed solvent is gradually improved, the extraction and dissolution of active substances with different polarities can be promoted in a longer time, the extraction effect of the active substances is greatly improved, and meanwhile, the obtained active substances containing CO ₂ The mixed solvent of (1), passing through a heating partThe state can be returned to the original state, which is beneficial to repeated use. The water phase contains impurities such as polysaccharide, protein, peptide, oligosaccharide and the like, the mixed solvent mainly contains active substances, and the invention further selects 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Extracting with nanometer magnetic fluid, performing reverse extraction on active substance, allowing the active substance to interact with anions and cations of 1-hydroxyethyl-3-methylimidazolyl group, allowing pi-pi interaction between benzene-containing compound and ionic liquid cation to realize extraction, adding into organic solvent, and adding 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ The active substances adsorbed by the nano magnetic fluid are extracted reversely to obtain extract liquor;

mixing the extract with the extraction oil, purifying by ceramic membrane, and further removing macromolecular impurities such as polysaccharide, protein, etc. to obtain high-purity essential oil mixture;

the peppermint essential oil is rich in levomenthol, isomenthone, pulegone, decyl acetate, menthyl acetate, methyl benzoate, alpha-and beta-pinene, beta-thujene, 3-pentanol, 2-hexanol, 3-octanol, dextromyrcene, limonene, eucalyptol and alpha-terpineol; the eucalyptus oil is rich in eucalyptol, linarene, camphor, anethole terpene and pomelo oil terpene; the lavender essential oil is rich in linalyl acetate, linalool, lavender alcohol, lavender acetate ester, p-1-menthene-4-ol, terpineol, caryophyllene and alloocimene; the clove essential oil contains rich eugenol, acetyl eugenol, beta-caryophyllene, alpha-humulene, methyl salicylate, caryophyllene oxide and piperitol; the melissa officinalis essential oil contains rich monoterpene aldehyde, wherein the content of citral is high, and neral and geranial are citral; the rosemary essential oil contains rich a-pinene, borneol, b-pinene, camphor, bornyl acetate, camphene, 1,8-cineole and limonene; the patchouli essential oil contains rich patchouli alkene, patchouli alcohol, patchouli ketone and patchouli oil which mainly contain methyl black pepper phenol and the like. The prepared mixed oil is adsorbed in the porous silica microspheres modified by polydopamine, and the silica microspheres contain a large number of tortuous pores and caves, so that the loading capacity of the loaded essential oil is improved, and the adsorption of the essential oil is also beneficial to the slow release of active volatile components of the essential oil.

The multifunctional plant essential oil composition prepared by the invention has the effects of refreshing, restoring consciousness, inducing resuscitation, relieving depression, relieving viscera, relaxing people, enabling people to relax, enabling people to feel joyful, preventing cold, smoothing nasal obstruction, relieving dizziness and headache, stopping dizziness and carsickness, preventing mosquito bites and mosquito bites, relieving swelling and itching after biting, promoting blood circulation to remove blood stasis, dredging channels and collaterals, effectively treating traumatic injury and the like.

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 description of the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a SEM image of the cross-section of the multifunctional essential oil microsphere prepared in example 1.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. 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.

Cellulase (5000U/g), pectinase (2500U/g), available from Xia Cheng (Beijing) Biotechnology development, inc.; the saccharomyces cerevisiae is purchased from Rihua biological koji making Limited company in the Longmarten Tan area of Luzhou city, and the strain number is more than or equal to 200 hundred million/g; bacillus subtilis with the strain number more than or equal to 200 hundred million/g and purchased from Wuhan Ai Nuosen Biotechnology Limited company; lactobacillus plantarum 100cfu/g, purchased from Weifang Ruichi Biotechnology GmbH; 1,3-dimethylimidazole bis (trifluoromethanesulfonylimide) salt available from Kyoto pure technology, inc., trioctylmethylbis (trifluoromethanesulfonylimide) ammonium salt available from Cian Ji Yue Biotechnology, inc.

Example 1

The embodiment provides a preparation method of a multifunctional essential oil composition, which specifically comprises the following steps:

s1, pretreatment of plant materials: drying 1 weight part of mint, 1 weight part of eucalyptus, 3 weight parts of lavender, 2 weight parts of clove, 1 weight part of melissa officinalis, 3 weight parts of rosemary and 5 weight parts of patchouli by 1000W microwave for 1h, mixing, crushing, and sieving by a 60-mesh sieve to obtain plant material powder;

s2, extracting by using a mixed solvent: adding the plant material powder prepared in the step S1 into a mixed solvent, wherein the solid-liquid ratio of the plant material powder to the mixed solvent is 1 g/mL, performing ultrasonic-assisted extraction for 30min at 500W, filtering, keeping the solid, and removing the solvent from the filtrate under reduced pressure to obtain extracted oil;

the mixed solvent is a mixed solvent of water, ethanol, acetonitrile and petroleum ether, and the volume ratio of the water, the ethanol, the acetonitrile and the petroleum ether is 3;

s3, enzymolysis: adding 10 parts by weight of the solid obtained in the step S2 into 100 parts by weight of water, adding 1 part by weight of complex enzyme for enzymolysis, and inactivating the enzyme at 100 ℃ for 15min to obtain an enzymolysis product;

the compound enzyme is a mixture of cellulase and pectinase, and the mass ratio of the compound enzyme to the pectinase is 3:1;

s4, fermentation: adding activated saccharomyces cerevisiae, bacillus subtilis and lactobacillus plantarum into the enzymolysis product obtained in the step S3, wherein the inoculation amounts are 3%, 2% and 0.5%, respectively, fermenting and culturing for 24 hours at 37 ℃, sterilizing by ultraviolet rays, and filtering to obtain a fermentation product;

the activation method comprises mixing Saccharomyces cerevisiae and Bacillus subtilisRespectively inoculating Bacillus and Lactobacillus plantarum in Gao's medium, culturing at 37 deg.C for 12 hr at 50r/min to obtain Bacillus with a bacterial content of 10 ⁸ cfu/mL strain seed solution;

s5, extracting ionic liquid: adding 10 parts by weight of the fermentation product obtained in the step S4 into 15 parts by weight of the ionic liquid-acetonitrile mixed solution, and introducing CO ₂ Introducing CO at a rate of 0.5mL/min for 30min, oscillating, and stopping introducing CO ₂ Obtaining a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, adding 2 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating adsorption, magnetic separation, washing, drying and filtering to obtain extraction slurry; heating the high-extraction ionic liquid-acetonitrile mixed solution to 50 ℃, treating for 30min, and dissipating CO ₂ And returning to obtain the ionic liquid-acetonitrile mixed solution, wherein the ionic liquid-acetonitrile mixed solution can be repeatedly used.

The mixed solution of the ionic liquid and the acetonitrile is a mixed solution of the ionic liquid and the acetonitrile in a mass ratio of 5:1; the ionic liquid is 1,3-dimethyl imidazole bis (trifluoromethanesulfonimide) salt and trioctylmethyl bis (trifluoromethanesulfonimide) ammonium according to the mass ratio of 3: 2;

s6, reverse extraction of active substances: adding 2 parts by weight of the extraction slurry prepared in the step S5 into 3 parts by weight of dichloromethane, stirring and extracting, performing magnetic separation, and removing an organic solvent to obtain an extraction liquid;

s7, purification: uniformly mixing 3 parts by weight of the extracted oil obtained in the step S2 and 2 parts by weight of the extract obtained in the step S6, filtering by adopting a small-aperture ceramic membrane with the aperture of 2000D, collecting filtrate obtained after small molecule filtration, and concentrating by adopting an ultrafiltration membrane with the aperture of 30nm to obtain purified volatile oil liquid with the relative density of 1.4;

s8, preparing porous silicon dioxide microspheres: dissolving 10 parts by weight of methyl orthosilicate in 50 parts by weight of ethyl acetate to obtain an oil phase; dissolving 2 parts by weight of pore-foaming agent and 0.5 part by weight of tween-20 in 100 parts by weight of water to obtain a water phase; adding the oil phase into the water phase, emulsifying with SPG rapid membrane with aperture of 5000nm, adjusting pH to 8, heating to 50 deg.C, reacting and curing for 3h, centrifuging at 5000r/min for 15min, washing with deionized water, and drying at 70 deg.C for 2h to obtain porous silica microspheres; the pore-foaming agent is a mixture of a macroporous pore-foaming agent polyoxyethylene sorbitan fatty acid ester and a mesoporous pore-foaming agent PEO106-PPO70-PEO10, and the mass ratio is 5:3.

S9, preparation of the modified porous silicon dioxide microspheres: uniformly dispersing 20 parts by weight of the porous silica microspheres prepared in the step S8 in 100 parts by weight of water, adding 25 parts by weight of dopamine hydrochloride and 2 parts by weight of catalyst, heating to 45 ℃ for reaction for 2h, centrifuging at 5000r/min for 15min, washing with deionized water, and drying at 70 ℃ for 2h to obtain modified porous silica microspheres;

the catalyst is Co containing 3wt% ²⁺ Tris-HCl solution with pH value of 5;

s10, preparing the multifunctional essential oil composition: adding 10 parts by weight of the modified porous silica microspheres obtained in the step S9 into 5 parts by weight of the purified volatile oil obtained in the step S7, stirring and adsorbing for 1 hour, and filtering to obtain multifunctional essential oil microspheres, namely the multifunctional essential oil composition, wherein the sectional SEM image of the multifunctional essential oil microspheres shown in the figure 1 shows that the microspheres form a large number of pores with uneven sizes, so that the adsorption and the slow release of the essential oil are facilitated.

Example 2

s1, pretreatment of plant materials: drying 3 parts by weight of mint, 2 parts by weight of eucalyptus, 5 parts by weight of lavender, 3 parts by weight of clove, 2 parts by weight of melissa officinalis, 5 parts by weight of rosemary and 7 parts by weight of patchouli by 1500W microwave for 2 hours, mixing, crushing, and sieving with a 100-mesh sieve to obtain plant material powder;

s2, mixed solvent extraction: adding the plant material powder prepared in the step S1 into a mixed solvent, wherein the solid-liquid ratio of the plant material powder to the mixed solvent is 1;

the mixed solvent is a mixed solvent of water, ethanol, acetonitrile and petroleum ether, and the volume ratio of the water, the ethanol, the acetonitrile and the petroleum ether is 5;

s3, enzymolysis: adding 10 parts by weight of the solid obtained in the step S2 into 100 parts by weight of water, adding 2 parts by weight of complex enzyme for enzymolysis, and inactivating the enzyme at 105 ℃ for 20min to obtain an enzymolysis product;

the compound enzyme is a mixture of cellulase and pectinase, and the mass ratio of the compound enzyme to the pectinase is 5:1;

s4, fermentation: adding activated saccharomyces cerevisiae, bacillus subtilis and lactobacillus plantarum into the enzymolysis product obtained in the step S3, wherein the inoculation amounts are 5%, 4% and 1.5%, respectively, fermenting and culturing for 48h at 40 ℃, sterilizing by ultraviolet rays, and filtering to obtain a fermentation product;

the activation method comprises respectively inoculating Saccharomyces cerevisiae, bacillus subtilis and Lactobacillus plantarum in Gao's medium, culturing at 39 deg.C for 18 hr at 70r/min to obtain extract with bacteria content of 10 ⁹ cfu/mL strain seed solution;

s5, extracting ionic liquid: adding 10 parts by weight of the fermentation product obtained in the step S4 into 20 parts by weight of the ionic liquid-acetonitrile mixed solution, and introducing CO ₂ Introducing CO at a rate of 1mL/min for 50min, oscillating, and stopping introducing CO ₂ Obtaining a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, adding 3 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating and adsorbing, magnetically separating, washing, drying and filtering to obtain extraction slurry; heating the high-extraction ionic liquid-acetonitrile mixed solution to 60 ℃, treating for 50min, and dissipating CO ₂ And returning to obtain the ionic liquid-acetonitrile mixed solution, wherein the ionic liquid-acetonitrile mixed solution can be repeatedly used.

The ionic liquid-acetonitrile mixed solution is a mixed solution of ionic liquid and acetonitrile in a mass ratio of 7:2; the ionic liquid is 1,3-dimethyl imidazole bis (trifluoromethyl) sulfonyl imide salt and trioctylmethyl bis (trifluoromethyl) sulfonyl imide ammonium according to the mass ratio of 5: 2;

s6, reverse extraction of active substances: adding 2 parts by weight of the extraction slurry prepared in the step S5 into 5 parts by weight of chloroform, stirring and extracting, performing magnetic separation, and removing an organic solvent to obtain an extraction liquid;

s7, purification: mixing 5 parts by weight of the extracted oil obtained in the step S2 and 7 parts by weight of the extract obtained in the step S6 uniformly, filtering by adopting a small-pore-diameter ceramic membrane with a pore diameter of 3000D, collecting filtrate obtained after small molecule filtration, and concentrating by adopting an ultrafiltration membrane with a pore diameter of 100nm to obtain purified volatile oil liquid with a relative density of 1.7;

s8, preparing porous silicon dioxide microspheres: dissolving 10 parts by weight of ethyl orthosilicate in 50 parts by weight of methyl acetate to obtain an oil phase; dissolving 3 parts by weight of pore-foaming agent and 1 part by weight of tween-60 in 100 parts by weight of water to obtain a water phase; adding the oil phase into the water phase, emulsifying by using an SPG rapid membrane with the aperture of 7000nm, adjusting the pH value to 9, heating to 70 ℃, reacting and curing for 5h, centrifuging for 15min at 5000r/min, washing by using deionized water, and drying for 2h at 70 ℃ to obtain the porous silicon dioxide microspheres; the pore-foaming agent is a mixture of a macroporous pore-foaming agent polyoxyethylene sorbitan fatty acid ester and a mesoporous pore-foaming agent PEO106-PPO70-PEO10, and the mass ratio is 7:3.

S9, preparation of the modified porous silicon dioxide microspheres: uniformly dispersing 20 parts by weight of the porous silica microspheres prepared in the step S8 in 100 parts by weight of water, adding 32 parts by weight of dopamine hydrochloride and 3 parts by weight of catalyst, heating to 55 ℃, reacting for 4h, centrifuging for 15min at 5000r/min, washing with deionized water, and drying for 2h at 70 ℃ to obtain modified porous silica microspheres;

the catalyst is Co containing 5wt% ²⁺ Tris-HCl solution with pH value of 6;

s10, preparing a multifunctional essential oil composition: and (3) adding 15 parts by weight of the modified porous silica microspheres obtained in the step (S9) into 7 parts by weight of the purified volatile oil liquid obtained in the step (S7), stirring and adsorbing for 2 hours, and filtering to obtain multifunctional essential oil microspheres, namely the multifunctional essential oil composition.

Example 3

s1, pretreatment of plant materials: drying 2 parts by weight of mint, 1.5 parts by weight of eucalyptus, 4 parts by weight of lavender, 2.5 parts by weight of clove, 1.5 parts by weight of melissa officinalis, 4 parts by weight of rosemary and 6 parts by weight of patchouli by 1250W microwave for 1.5h, mixing, crushing, and sieving by a 80-mesh sieve to obtain plant material powder;

s2, extracting by using a mixed solvent: adding the plant material powder prepared in the step S1 into a mixed solvent, wherein the solid-liquid ratio of the plant material powder to the mixed solvent is 1 g/mL-4 g/mL, performing ultrasonic-assisted extraction for 40min at 600W, filtering, keeping the solid, and removing the solvent from the filtrate under reduced pressure to obtain extracted oil;

the mixed solvent is a mixed solvent of water, ethanol, acetonitrile and petroleum ether, and the volume ratio of the water, the ethanol, the acetonitrile and the petroleum ether is 4;

s3, enzymolysis: adding 10 parts by weight of the solid obtained in the step S2 into 100 parts by weight of water, adding 1.5 parts by weight of complex enzyme for enzymolysis, and inactivating the enzyme at 105 ℃ for 20min to obtain an enzymolysis product;

the compound enzyme is a mixture of cellulase and pectinase, and the mass ratio of the compound enzyme to the pectinase is 4:1;

s4, fermentation: adding activated saccharomyces cerevisiae, bacillus subtilis and lactobacillus plantarum into the enzymolysis product obtained in the step S3, wherein the inoculation amounts are 4%, 3% and 1%, respectively, fermenting and culturing for 36 hours at 38 ℃, sterilizing by ultraviolet rays, and filtering to obtain a fermentation product;

the activation method comprises respectively inoculating Saccharomyces cerevisiae, bacillus subtilis and Lactobacillus plantarum in Gao's medium, culturing at 38 deg.C for 15 hr to obtain extract with bacteria content of 10 ⁹ cfu/mL strain seed solution;

s5, extracting ionic liquid: adding 10 parts by weight of the fermentation product obtained in the step S4 into 17 parts by weight of the ionic liquid-acetonitrile mixed solution, and introducing CO ₂ Introducing CO at a rate of 0.7mL/min for 40min, oscillating, and stopping introducing CO ₂ To obtain a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, adding 2.5 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating and adsorbing, magnetically separating, washing, drying and filtering to obtain extraction slurry; heating the high-extraction ionic liquid-acetonitrile mixed solution to 55 ℃, and treating for 40minAfter that, CO is escaped ₂ And returning to obtain the ionic liquid-acetonitrile mixed solution, wherein the ionic liquid-acetonitrile mixed solution can be repeatedly used.

The ionic liquid-acetonitrile mixed solution is a mixed solution of ionic liquid and acetonitrile according to a mass ratio of 6; the ionic liquid is 1,3-dimethyl imidazole bis (trifluoromethanesulfonimide) salt and trioctylmethyl bis (trifluoromethanesulfonimide) ammonium according to the mass ratio of 4: 2;

s6, reverse extraction of active substances: adding 2 parts by weight of the extraction slurry prepared in the step S5 into 4 parts by weight of dichloromethane, stirring and extracting, performing magnetic separation, and removing an organic solvent to obtain an extraction liquid;

s7, purification: uniformly mixing 4 parts by weight of the extracted oil obtained in the step S2 and 5 parts by weight of the extract obtained in the step S6, filtering by adopting a small-aperture ceramic membrane with the aperture of 2500D, collecting filtrate obtained after small molecule filtration, and concentrating by adopting an ultrafiltration membrane with the aperture of 60nm to obtain purified volatile oil liquid with the relative density of 1.55;

s8, preparing porous silica microspheres: dissolving 10 parts by weight of ethyl orthosilicate in 50 parts by weight of dichloromethane to obtain an oil phase; dissolving 2.5 parts by weight of pore-foaming agent and 0.7 part by weight of tween-80 in 100 parts by weight of water to obtain a water phase; adding the oil phase into the water phase, emulsifying with an SPG rapid membrane with the pore diameter of 6000nm, adjusting the pH value to 8.5, heating to 60 ℃, reacting and curing for 4h, centrifuging for 15min at 5000r/min, washing with deionized water, and drying at 70 ℃ for 2h to obtain the porous silicon dioxide microspheres; the pore-foaming agent is a mixture of a macroporous pore-foaming agent polyoxyethylene sorbitan fatty acid ester and a mesoporous pore-foaming agent PEO106-PPO70-PEO10, and the mass ratio is 6:3.

S9, preparation of the modified porous silicon dioxide microspheres: uniformly dispersing 20 parts by weight of the porous silica microspheres prepared in the step S8 in 100 parts by weight of water, adding 29 parts by weight of dopamine hydrochloride and 2.5 parts by weight of catalyst, heating to 50 ℃, reacting for 3h, centrifuging for 15min at 5000r/min, washing with deionized water, and drying for 2h at 70 ℃ to obtain modified porous silica microspheres;

the catalyst is Co containing 4wt% ²⁺ Tris-HCl solution of pH 5.5；

S10, preparing a multifunctional essential oil composition: and (3) adding 12 parts by weight of the modified porous silica microspheres obtained in the step (S9) into 6 parts by weight of the purified volatile oil liquid obtained in the step (S7), stirring and adsorbing for 1.5 hours, and filtering to obtain multifunctional essential oil microspheres, namely the multifunctional essential oil composition.

Example 4

Compared with the ionic liquid in the example 3, the ionic liquid is single 1,3-dimethyl imidazole bis (trifluoromethanesulfonimide) salt, and other conditions are not changed.

Example 5

Compared with the ionic liquid in the example 3, the ionic liquid is single trioctylmethylbistrifluoromethanesulfonylimide ammonium, and other conditions are not changed.

Example 6

Compared with the embodiment 3, the pore-foaming agent is single macroporous pore-foaming agent polyoxyethylene sorbitan fatty acid ester, and other conditions are not changed.

Example 7

Compared with example 3, the pore-forming agent is a single mesoporous pore-forming agent PEO106-PPO70-PEO10, and other conditions are not changed.

Example 8

Compared with the embodiment 3, the mixed solvent is a mixed solvent of ethanol, acetonitrile and petroleum ether, the volume ratio of the ethanol, the acetonitrile and the petroleum ether is 10.

Example 9

Compared with the embodiment 3, the mixed solvent is a mixed solvent of water, acetonitrile and petroleum ether, the volume ratio of the water, the acetonitrile and the petroleum ether is 10.

Example 10

Compared with the embodiment 3, the mixed solvent is the mixed solvent of water and ethanol, the volume ratio of the water to the ethanol is 4:6, and other conditions are not changed.

Example 11

Compared with the example 3, the complex enzyme is single cellulase, and other conditions are not changed.

Example 12

Compared with the example 3, the compound enzyme is single pectinase, and other conditions are not changed.

Comparative example 1

Compared with the embodiment 3, the microwave drying in the step S1 is changed into the vacuum drying, and other conditions are not changed.

The method comprises the following specific steps:

s1, pretreatment of plant materials: vacuum drying herba Menthae 2 weight parts, eucalyptus 1.5 weight parts, lavender 4 weight parts, flos Caryophylli 2.5 weight parts, melissa officinalis 1.5 weight parts, herba Rosmarini officinalis 4 weight parts, and herba Agastaches 6 weight parts at 70 deg.C for 1.5h, mixing, pulverizing, and sieving with 80 mesh sieve to obtain plant material powder.

Comparative example 2

Compared with example 3, the step of extracting the mixed solvent of step S2 was not performed, and other conditions were not changed.

The method comprises the following specific steps:

s1, pretreatment of plant materials: drying 2 parts by weight of mint, 1.5 parts by weight of eucalyptus, 4 parts by weight of lavender, 2.5 parts by weight of clove, 1.5 parts by weight of melissa officinalis, 4 parts by weight of rosemary and 6 parts by weight of pogostemon cablin by 1250W under microwave for 1.5h, mixing, crushing and sieving by a 80-mesh sieve to obtain plant material powder;

s2, enzymolysis: adding 10 parts by weight of the plant material powder prepared in the step S1 into 100 parts by weight of water, adding 1.5 parts by weight of complex enzyme for enzymolysis, and inactivating the enzyme at 105 ℃ for 20min to obtain an enzymolysis product;

s3, fermentation: adding activated saccharomyces cerevisiae, bacillus subtilis and lactobacillus plantarum into the enzymolysis product obtained in the step S2, wherein the inoculation amounts are 4%, 3% and 1%, respectively, fermenting and culturing for 36h at 38 ℃, sterilizing by ultraviolet rays, and filtering to obtain a fermentation product;

s4, ionic liquid extraction: 10 parts by weight of the fermentation product obtained in step S3Adding the mixture into a mixed solution of 17 parts by weight of ionic liquid and acetonitrile, and introducing CO ₂ Introducing CO at a rate of 0.7mL/min for 40min, oscillating, and stopping introducing CO ₂ To obtain a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, and adding 2.5 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating adsorption, magnetic separation, washing, drying and filtering to obtain extraction slurry; heating the high-extraction ionic liquid-acetonitrile mixed solution to 55 ℃, and dissipating CO after 40min of treatment ₂ And returning to obtain the ionic liquid-acetonitrile mixed solution, wherein the ionic liquid-acetonitrile mixed solution can be repeatedly used.

s5, reverse extraction of active substances: adding 2 parts by weight of the extraction slurry prepared in the step S4 into 4 parts by weight of dichloromethane, stirring and extracting, performing magnetic separation, and removing the organic solvent to obtain an extraction liquid;

s6, purification: mixing 9 parts by weight of the extract obtained in the step S5 uniformly, filtering by adopting a small-aperture ceramic membrane with the aperture of 2500D, collecting filtrate after small molecule filtration, and concentrating by adopting an ultrafiltration membrane with the aperture of 60nm to obtain purified volatile oil liquid with the relative density of 1.55;

s7, preparing porous silicon dioxide microspheres: dissolving 10 parts by weight of ethyl orthosilicate in 50 parts by weight of dichloromethane to obtain an oil phase; dissolving 2.5 parts by weight of pore-foaming agent and 0.7 part by weight of tween-80 in 100 parts by weight of water to obtain a water phase; adding the oil phase into the water phase, emulsifying by using an SPG rapid membrane with the pore diameter of 6000nm, adjusting the pH value to 8.5, heating to 60 ℃, reacting and curing for 4h, centrifuging at 5000r/min for 15min, washing by using deionized water, and drying at 70 ℃ for 2h to obtain porous silica microspheres; the pore-foaming agent is a mixture of a macroporous pore-foaming agent polyoxyethylene sorbitan fatty acid ester and a mesoporous pore-foaming agent PEO106-PPO70-PEO10, and the mass ratio is 6:3.

S8, preparing the modified porous silicon dioxide microspheres: uniformly dispersing 20 parts by weight of the porous silica microspheres prepared in the step S7 in 100 parts by weight of water, adding 29 parts by weight of dopamine hydrochloride and 2.5 parts by weight of catalyst, heating to 50 ℃, reacting for 3h, centrifuging for 15min at 5000r/min, washing with deionized water, and drying for 2h at 70 ℃ to obtain modified porous silica microspheres;

the catalyst is Co containing 4wt% ²⁺ Tris-HCl solution of pH 5.5;

s9, preparing the multifunctional essential oil composition: and (3) adding 12 parts by weight of the modified porous silica microspheres obtained in the step (S8) into 6 parts by weight of the purified volatile oil liquid obtained in the step (S6), stirring and adsorbing for 1.5 hours, and filtering to obtain multifunctional essential oil microspheres, namely the multifunctional essential oil composition.

Comparative example 3

Compared with the embodiment 3, the step of S3 enzymolysis is not carried out, and other conditions are not changed.

The method comprises the following specific steps:

s2, mixed solvent extraction: adding the plant material powder prepared in the step S1 into a mixed solvent, wherein the solid-liquid ratio of the plant material powder to the mixed solvent is 1 g/mL-4 g/mL, performing ultrasonic-assisted extraction for 40min at 600W, filtering, keeping the solid, and removing the solvent from the filtrate under reduced pressure to obtain extracted oil;

s3, fermentation: adding 10 parts by weight of the solid obtained in the step S2 into 100 parts by weight of water, adding activated saccharomyces cerevisiae, bacillus subtilis and lactobacillus plantarum, wherein the inoculation amounts are 4%, 3% and 1%, respectively, fermenting and culturing for 36 hours at 38 ℃, sterilizing by ultraviolet rays, and filtering to obtain a fermentation product;

s4, ionic liquid extraction: adding 10 parts by weight of the fermentation product obtained in the step S3 into 17 parts by weight of the ionic liquid-acetonitrile mixed solution, and introducing CO ₂ Introducing CO at a rate of 0.7mL/min for 40min, oscillating, and stopping introducing CO ₂ To obtain a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, adding 2.5 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating adsorption, magnetic separation, washing, drying and filtering to obtain extraction slurry; heating the high-extraction ionic liquid-acetonitrile mixed solution to 55 ℃, and dissipating CO after 40min of treatment ₂ And returning to obtain the ionic liquid-acetonitrile mixed solution, wherein the ionic liquid-acetonitrile mixed solution can be repeatedly used.

s6, purification: mixing 4 parts by weight of the extracted oil obtained in the step S2 and 5 parts by weight of the extract obtained in the step S5 uniformly, filtering by adopting a small-pore-diameter ceramic membrane with the pore diameter of 2500D, collecting filtrate obtained after small molecule filtration, and concentrating by adopting an ultrafiltration membrane with the pore diameter of 60nm to obtain purified volatile oil liquid with the relative density of 1.55;

s7, preparing porous silicon dioxide microspheres: dissolving 10 parts by weight of ethyl orthosilicate in 50 parts by weight of dichloromethane to obtain an oil phase; dissolving 2.5 parts by weight of pore-foaming agent and 0.7 part by weight of tween-80 in 100 parts by weight of water to obtain a water phase; adding the oil phase into the water phase, emulsifying with an SPG rapid membrane with the pore diameter of 6000nm, adjusting the pH value to 8.5, heating to 60 ℃, reacting and curing for 4h, centrifuging for 15min at 5000r/min, washing with deionized water, and drying at 70 ℃ for 2h to obtain the porous silicon dioxide microspheres; the pore-foaming agent is a mixture of a macroporous pore-foaming agent polyoxyethylene sorbitan fatty acid ester and a mesoporous pore-foaming agent PEO106-PPO70-PEO10, and the mass ratio is 6:3.

the catalyst is Co containing 4wt% ²⁺ The Tris-HCl solution of pH 5.5;

Comparative example 4

In step S1, mint was not added, and other conditions were not changed, as compared with example 3.

The method comprises the following specific steps:

s1, pretreatment of plant materials: drying 2 parts by weight of eucalyptus, 5 parts by weight of lavender, 3 parts by weight of clove, 2 parts by weight of melissa officinalis, 5 parts by weight of rosemary and 7 parts by weight of patchouli by 1500W microwave for 2h, mixing, crushing, and sieving with a 100-mesh sieve to obtain the plant material powder.

Comparative example 5

Compared with example 3, no eucalyptus was added in step S1, and other conditions were not changed.

The method comprises the following specific steps:

s1, pretreatment of plant materials: drying 3 parts by weight of mint, 5 parts by weight of lavender, 3 parts by weight of clove, 2 parts by weight of melissa officinalis, 5 parts by weight of rosemary and 7 parts by weight of patchouli by 1500W microwave for 2h, mixing, crushing, and sieving with a 100-mesh sieve to obtain the plant material powder.

Comparative example 6

Compared with the example 3, the lavender is not added in the step S1, and other conditions are not changed.

The method comprises the following specific steps:

s1, pretreatment of plant materials: drying herba Menthae 3 weight parts, eucalyptus globulus 2 weight parts, flos Caryophylli 3 weight parts, herba Melissae axillaris 2 weight parts, herba Rosmarini officinalis 5 weight parts, and herba Agastaches 7 weight parts under 1500W microwave for 2 hr, mixing, pulverizing, and sieving with 100 mesh sieve to obtain plant material powder.

Comparative example 7

In comparison with example 3, the fermentation step of step S4 was not performed, and other conditions were not changed.

The method comprises the following specific steps:

s4, ionic liquid extraction: adding 10 parts by weight of the enzymolysis product prepared in the step S3 into 17 parts by weight of ionic liquid-acetonitrile mixed solution, and introducing CO ₂ The flow rate is 0.7mL/min, the flow time is 40min, and the mixture is vibratedStopping introducing CO ₂ To obtain a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, adding 2.5 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating adsorption, magnetic separation, washing, drying and filtering to obtain extraction slurry; heating the high-extraction ionic liquid-acetonitrile mixed solution to 55 ℃, treating for 40min, and dissipating CO ₂ And returning to obtain the ionic liquid-acetonitrile mixed solution, wherein the ionic liquid-acetonitrile mixed solution can be repeatedly used.

s6, purification: mixing 4 parts by weight of the extracted oil obtained in the step S2 and 5 parts by weight of the extract obtained in the step S5 uniformly, filtering by adopting a small-aperture ceramic membrane with the aperture of 2500D, collecting filtrate obtained after small molecule filtration, and concentrating by adopting an ultrafiltration membrane with the aperture of 60nm to obtain purified volatile oil liquid with the relative density of 1.55;

the catalyst is Co containing 4wt% ²⁺ Tris-HCl solution of pH 5.5;

Comparative example 8

In comparison with example 3, no CO was introduced in step S5 ₂ Other conditions are not changed.

The method comprises the following specific steps:

s5, extracting ionic liquid: adding 10 parts by weight of the fermentation product obtained in the step S4 into 17 parts by weight of ionic liquid-acetonitrile mixed solution, stirring and extracting, standing for layering, separating liquid, removing water layer, adding 2.5 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating adsorption, magnetic separation, washing, drying and filtering to obtain extraction slurry;

the ionic liquid-acetonitrile mixed solution is a mixed solution of ionic liquid and acetonitrile according to a mass ratio of 6; the ionic liquid is 1,3-dimethyl imidazole bis (trifluoromethanesulfonimide) salt and trioctylmethyl bis (trifluoromethanesulfonimide) ammonium according to the mass ratio of 4:2, or a mixture thereof.

Comparative example 9

Compared with the example 3, the ionic liquid-acetonitrile mixed solution in the step S5 is replaced by acetonitrile, and other conditions are not changed.

The method comprises the following specific steps:

s5, extracting ionic liquid: adding 10 parts by weight of the fermentation product obtained in the step S4 into 17 parts by weight of acetonitrile, and introducing CO ₂ Introducing CO at the rate of 0.7mL/min for 40min, oscillating, and stopping introducing CO ₂ Standing and separatingLayer separation, removal of the aqueous layer, addition of 2.5 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillation adsorption, magnetic separation, washing, drying and filtering to obtain the extraction slurry.

Comparative example 10

Compared with example 3, the ionic liquid extraction step of step S5 was not performed, and other conditions were not changed.

The method comprises the following specific steps:

s5, reverse extraction of active substances: adding 2 parts by weight of the fermentation product obtained in the step S4 into 4 parts by weight of dichloromethane, stirring and extracting, performing magnetic separation, and removing the organic solvent to obtain an extract liquid;

s7, preparing the porous silica microspheres: dissolving 10 parts by weight of ethyl orthosilicate in 50 parts by weight of dichloromethane to obtain an oil phase; dissolving 2.5 parts by weight of pore-foaming agent and 0.7 part by weight of tween-80 in 100 parts by weight of water to obtain a water phase; adding the oil phase into the water phase, emulsifying with an SPG rapid membrane with the pore diameter of 6000nm, adjusting the pH value to 8.5, heating to 60 ℃, reacting and curing for 4h, centrifuging for 15min at 5000r/min, washing with deionized water, and drying at 70 ℃ for 2h to obtain the porous silicon dioxide microspheres; the pore-foaming agent is a mixture of a macroporous pore-foaming agent polyoxyethylene sorbitan fatty acid ester and a mesoporous pore-foaming agent PEO106-PPO70-PEO10, and the mass ratio is 6:3.

the catalyst is Co containing 4wt% ²⁺ The Tris-HCl solution of pH 5.5;

Comparative example 11

Compared with the embodiment 3, no pore-foaming agent is added in the step S8, and other conditions are not changed.

The method comprises the following specific steps:

s8, preparation of the silicon dioxide microspheres: dissolving 10 parts by weight of ethyl orthosilicate in 50 parts by weight of dichloromethane to obtain an oil phase; dissolving 0.7 part by weight of tween-80 in 100 parts by weight of water to obtain a water phase; adding the oil phase into the water phase, emulsifying with SPG rapid membrane with pore diameter of 6000nm, adjusting pH to 8.5, heating to 60 deg.C, reacting and curing for 4h, centrifuging at 5000r/min for 15min, washing with deionized water, and drying at 70 deg.C for 2h to obtain silicon dioxide microsphere.

Comparative example 12

Step S9 was not performed, and other conditions were not changed, as compared with example 3.

The method comprises the following specific steps:

the activating method comprises the following steps of using saccharomyces cerevisiaeRespectively inoculating the bacillus subtilis and the lactobacillus plantarum in a Gao's medium, culturing at 38 ℃ for 60r/min for 15h to obtain the bacillus with the bacterial content of 10 ⁹ cfu/mL strain seed solution;

s5, ionic liquid extraction: adding 10 parts by weight of the fermentation product obtained in the step S4 into 17 parts by weight of the ionic liquid-acetonitrile mixed solution, and introducing CO ₂ Introducing CO at the rate of 0.7mL/min for 40min, oscillating, and stopping introducing CO ₂ To obtain a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, adding 2.5 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating and adsorbing, magnetically separating, washing, drying and filtering to obtain extraction slurry; heating the high-extraction ionic liquid-acetonitrile mixed solution to 55 ℃, treating for 40min, and dissipating CO ₂ And returning to obtain the ionic liquid-acetonitrile mixed solution, wherein the ionic liquid-acetonitrile mixed solution can be repeatedly used.

S9, preparing the multifunctional essential oil composition: and (3) adding 12 parts by weight of the porous silica microspheres obtained in the step (S8) into 6 parts by weight of the purified volatile oil liquid obtained in the step (S7), stirring and adsorbing for 1.5 hours, and filtering to obtain multifunctional essential oil microspheres, namely the multifunctional essential oil composition.

Test example 1 sustained Release Performance test

The multifunctional essential oil compositions prepared in examples 1 to 3, examples 6 and 7 and comparative examples 11 to 12 of the present invention were subjected to release property studies.

Storing the same mass of origanum essential oil microcapsules for 30 days at 25 ℃ under aerobic condition, measuring the release rate of the origanum essential oil every 5 days, and repeating the measurement for 3 times. The release rate is calculated according to the following formula:

release rate (%) = (V) ₁ -V _t ）/V ₁ ×100%

In the formula: v ₁ Is the initial microsphere mass/g; v _t Day t microsphere mass/g.

The results are shown in Table 1.

TABLE 1

As can be seen from the table above, the multifunctional essential oil microspheres prepared in the embodiments 1-3 of the invention can adsorb essential oil in the pore canals of the microspheres, and have a long-acting slow-release effect.

Compared with the embodiment 3, the pore-forming agent is single macroporous pore-forming agent polyoxyethylene sorbitan fatty acid ester or mesoporous pore-forming agent PEO106-PPO70-PEO10, the slow release effect is reduced, and the release rate is improved at the same time. Compared with the example 3, the comparative example 11 has the advantages that the pore-foaming agent is not added in the step S8, the slow release effect is reduced, and the release rate is obviously improved at the same time. Under the action of the macroporous pore-foaming agent and the mesoporous pore-foaming agent, the silicon dioxide microspheres can form complex and compact pore channels, so that the plant essential oil can be fully adsorbed and slowly released in the later period.

Compared with the example 3, the slow release effect is reduced without the step S9, the release rate is obviously improved at the same time, the surface and the interior of the modified polydopamine contain abundant polydopamine substances, and the abundant hydroxyl, carboxyl, amino and phenyl of the polydopamine are beneficial to forming hydrogen bonds with active substances in essential oil, such as alcohol, ketone, acid, ester and the like, so that the active substances are stably adsorbed, meanwhile, the phenyl is easy to form pi-pi conjugation with aromatic groups in the active substances, such as methyl salicylate and the like, so that the phenyl is stably fixed in the silicon dioxide microspheres, and the loading capacity and the slow release effect of the microspheres on the active substances in the essential oil are improved.

Test example 2 bacteriostatic test

And (4) judging the inhibition effect of the sample to be tested on escherichia coli (ATCC 25922), staphylococcus aureus (ATCC 6538) and candida albicans (ATCC 10231) through a bacteriostasis zone test.

Pouring the sterilized culture medium into sterile culture dishes with a diameter of 9cm on a sterile clean bench, allowing each dish to have 15-20mL of the culture medium, cooling, and adding 0.2mL of a suspension of Escherichia coli (ATCC 25922), staphylococcus aureus (ATCC 6538) and Candida albicans (ATCC 10231) with a concentration of 10 ⁶ cfu/mL), the surface of the culture medium is evenly coated, an Oxford cup is vertically placed on the surface of the corresponding culture medium, the oxford cup is pressed to be in contact with the culture medium without gaps, 3 small tubes are placed on each flat plate, 0.1mg of the multifunctional essential oil composition prepared in the examples 1-12 and the comparative examples 1-12 is respectively dripped into each small tube, the marking is carried out, and then the multifunctional essential oil composition is placed in an incubator at 37 ℃ for 72 hours, and the result is observed. During the cultivation, the test fungus constantly grows, and the appearance that awaits measuring of interpolation uses the oxford cup as the original point to spread all around, forms the circular region that the bacterial colony can not grow, is "antibacterial circle", and after the experiment, test "antibacterial circle" diameter.

The results are shown in Table 2.

TABLE 2

As can be seen from the above table, the multifunctional essential oil compositions prepared in the embodiments 1 to 3 of the present invention have good bacteriostatic properties.

Test example 3 experiments on fullness

The experiments were performed with reference to NY/T1151.2-2006 standard for mite killing and acaricidal agent testing. The multifunctional essential oil compositions prepared in examples 1 to 12 and comparative examples 1 to 12 were divided into 24 groups, and for each group of samples, 4 petri dishes were taken, 3 of which were added with 10g of the sample to be tested. The other dish was not filled with the test sample as a negative control. The upper edge of the inner wall of each culture dish is evenly coated with white oil and vaseline mixture. 200 test insects are put in the center of each culture dish, 0.05g of mite feed is put in the center of each culture dish when 30min is needed, and then the culture dish is placed in a water-proof incubator. The dead mites were examined and recorded at 48 h. The average of 3 dishes in each group was taken as the final result.

The mite killing rate is calculated as follows:

mite control rate (%) = (1-test insect number of experimental group/test insect number of negative control group) × 100%

The results are shown in Table 3.

TABLE 3

As can be seen from the above table, the multifunctional essential oil compositions prepared in examples 1-3 of the present invention have good mite-killing effect.

Test example 4 mosquito repellent effect test

10g of the multifunctional essential oil composition prepared in examples 1 to 12 of the present invention and comparative examples 1 to 12 was added to 20mL of a 20wt% sodium alginate solution, and 2mL of 5wt% calcium chloride was added dropwise to form a gel.

Selecting Aedes albopictus with qualified attack force and 4 or more test persons according to the requirements of GB/T13917.9-2009, and placing the test persons on the backs of the test personsThe skin of the test subject 5 cm X5 cm was identified and the gel prepared above was applied evenly to the back of the hand (at 1.5 mg/cm) ² ) The back of the hand was covered with a glove, exposing only the gel-coated 4 cm × 4 cm skin, and the other hand was blank. 2h, the hand coated with the gel is put into a mosquito cage for 2 min, and whether the mosquito stops falling and sucks blood is observed. And then testing every 1h, judging that the tested object is invalid as long as one mosquito stops and sucks blood during the testing period, and recording the effective protection time (h). Before each test, the reference hand is required to perform the test first, when the attack force of the mosquitoes is qualified, the test can be continuously performed, otherwise, the mosquitoes with qualified attack force are replaced to continuously perform the test.

Calculating the effective protection time: average number of effective protection times per group of test persons.

The results are shown in Table 4.

TABLE 4

As can be seen from the above table, the multifunctional essential oil composition prepared in embodiments 1 to 3 of the present invention has a good mosquito repelling effect.

Test example 5 anti-inflammatory test

162 SPF male mice are randomly divided into 27 groups, namely a model group, a blank group, a compound dexamethasone acetate cream group, examples 1-12 and comparative examples 1-12, wherein each group comprises 6 mice. Taking the test substance for external administration 2 times per day on the front and back sides of the auricle, 1.2g/kg each time, administering 0.2g/kg compound dexamethasone acetate cream group, administering 1.2g/kg matrix gel each time in blank group, and continuously administering for 7d. 0.5h after the last dose, each mouse was smeared on both sides of the right ear with 0.05mL of xylene, and the animals were sacrificed 30min later. Two ears are cut along the auricle of the mouse, ear pieces with the same parts and the same size are respectively chiseled out by a puncher with the diameter of 8mm, the ear pieces are weighed, the weight difference value of the two ear pieces is used as the swelling degree, and the swelling inhibition rate is calculated.

Swelling inhibition (%) = (average swelling rate in model group-average swelling rate in administration group)/average swelling rate in model group × 100%.

The results are shown in Table 5.

TABLE 5

As can be seen from the above table, the multifunctional essential oil compositions prepared in examples 1 to 3 of the present invention have excellent anti-inflammatory effects.

Compared with the ionic liquid in the embodiment 3, the ionic liquid is single 1,3-dimethyl imidazole bis (trifluoromethyl) sulfonyl imide salt or trioctylmethyl bis (trifluoromethyl) sulfonyl imide ammonium, and the antibacterial, anti-inflammatory, acaricidal and mosquito-repellent effects are reduced. Compared with the embodiment 3, the ionic liquid-acetonitrile mixed solution in the step S5 is replaced by acetonitrile, so that the antibacterial, anti-inflammatory, acaricidal and mosquito-repellent effects are obviously reduced. Compared with the example 3, the comparative example 10 has the advantages that the ionic liquid extraction step of the step S5 is not carried out, and the antibacterial, anti-inflammatory, mite-killing and mosquito-repelling effects are obviously reduced. The enzymolysis product is further extracted by the mixed solution of ionic liquid and acetonitrile, the enzymolysis product contains a large amount of polysaccharide, protein, peptide, oligosaccharide and other impurities, the ionic liquid is extracted by the mixed solution of ionic liquid and acetonitrile, the ionic liquid consists of positively charged ions and negatively charged ions, the ionic liquid is in a liquid state at the temperature of between 100 ℃ below zero and 200 ℃, the ionic liquid can not be steam to generate harmful gas causing air pollution generally, and the ionic liquid can be repeatedly used for a plurality of times. The hydrophobic ionic liquid is not dissolved with water, substances dissolved in water can be extracted by utilizing the principle of 'similarity and dissolution', and the ionic liquid-acetonitrile mixed solvent is used as an extracting agent for extraction and separation, so that the problems of high viscosity and high cost of the ionic liquid can be solved, and the extraction of active substances can be promoted, thereby achieving good effects of separating impurities and extracting the active substances.

Comparative example 8 in comparison with example 3, no CO was fed in step S5 ₂ The effects of bacteriostasis, inflammation diminishing, mite killing and mosquito repelling are reduced.In the extraction process, CO is slowly introduced ₂ The polarity of the mixed solvent is gradually improved, the extraction and dissolution of active substances with different polarities can be promoted in a longer time, the extraction effect of the active substances is greatly improved, and meanwhile, the obtained active substances containing CO ₂ The mixed solvent can return to the original state after being heated, and is favorable for repeated use.

Compared with the embodiment 3, the pore-forming agent is single macroporous pore-forming agent polyoxyethylene sorbitan fatty acid ester or mesoporous pore-forming agent PEO106-PPO70-PEO10 in the embodiments 6 and 7, and compared with the embodiment 3, the pore-forming agent is not added in the step S8, so that the bacteriostatic, anti-inflammatory, mite-killing and mosquito-repellent effects are reduced in the comparative example 11. According to the invention, a plurality of active compositions are mixed, and the prepared mixed oil is adsorbed in porous silica microspheres modified by polydopamine, and the silica microspheres contain a large number of tortuous pores and caves, so that the loading capacity of the loaded essential oil is improved, and meanwhile, the adsorption of the essential oil is also beneficial to the slow release of the active volatile components of the essential oil, so that the effect is exerted.

In examples 8, 9 and 10, compared with example 3, the mixed solvent is a mixed solvent of ethanol, acetonitrile and petroleum ether in sequence, wherein the volume ratio of the ethanol, the acetonitrile and the petroleum ether is 10; the mixed solvent of water, acetonitrile and petroleum ether, wherein the volume ratio of water, acetonitrile and petroleum ether is 10; the mixed solvent of water and ethanol, the volume ratio of water to ethanol is 4:6, and the effects of bacteriostasis, inflammation diminishing, mite killing and mosquito repelling are reduced. Compared with the example 3, the comparative example 2 has the advantages that the mixed solvent extraction step in the step S2 is not carried out, and the antibacterial, anti-inflammatory, acaricidal and mosquito-repellent effects are obviously reduced. The invention utilizes the mixed solvent of water, ethanol, acetonitrile and petroleum ether to extract under the action of ultrasonic wave, the four-phase system of the water-ethanol-acetonitrile-petroleum ether mixed solvent comprises polar solvents of water, ethanol, acetonitrile and the like, and simultaneously comprises nonpolar solvents of petroleum ether, and simultaneously, after microwave drying, the cells of the plant material powder are cracked and damaged, and are further extracted under the assistance of ultrasonic wave to be further cracked, thereby being beneficial to the dissolution of active substances and the similar dissolution of the active substances by the solvent, and being dissolved in the solvent, thereby improving the solubility of the active substances in the solvent and improving the extraction rate.

Compared with the embodiment 3, the embodiment 11 and 12 have the advantages that the complex enzyme is single cellulase or pectinase, and the anti-inflammation and mosquito-repellent effects are reduced. Compared with the embodiment 3, the step S3 of enzymolysis is not carried out, and the anti-inflammation and mosquito-repellent effects are obviously reduced. The solid obtained by filtering is subjected to enzymolysis by adopting the compound enzyme which is a mixture of cellulase and pectinase, so that the cell walls, pectin and the like of plant cells are subjected to enzymolysis, a large amount of active substances in the cells are further dissolved out, the extraction rate is improved, and the effect of the obtained essential oil composition is improved.

Compared with the embodiment 3, the microwave drying in the step S1 is changed into the vacuum drying, so that the anti-inflammation, acaricidal and mosquito-repellent effects are reduced. According to the invention, the components are firstly dried by microwave, compared with the traditional drying method, for example, the drying method can expose the plant materials in high-temperature air for a long time to cause the terpenes to be degraded into the monoterpenes, and simultaneously, the alcohols are esterified to form esters, so that the composition of part of essential oil at the later stage is changed. Vacuum drying significantly changes the type and amount of active ingredients of the essential oil due to long vacuum treatment. Infrared drying causes oxidation and chemical structural rearrangement, which results in the production of new volatile compounds that differ significantly from the structure of the plant material itself. The microwave drying method adopted in the invention can improve the internal temperature of the plant material to be higher, cause the damage of plant cells and the rupture of cell walls, and is beneficial to the release of volatile compounds.

In comparative examples 4, 5 and 6, compared with example 3, the step S1 without the mint, eucalyptus and lavender has reduced antibacterial, anti-inflammatory, acaricidal and mosquito-repellent effects. The mint essential oil is rich in levo-menthol, levo-menthone, isomenthone, pulegone, decyl acetate, menthyl acetate, methyl benzoate, alpha-and beta-pinene, beta-thujene, 3-pentanol, 2-hexanol, 3-octanol, dextro-myrcene, limonene, cineole and alpha-terpineol; the eucalyptus oil is rich in eucalyptol, linarene, camphor, anethole terpene and pomelo oil terpene; the lavender essential oil is rich in linalyl acetate, linalool, lavender alcohol, lavender ester acetate, p-1-menthene-4-ol, terpineol, caryophyllene, and alloocimene. The raw materials have synergistic effect, and the effects of refreshing brain, inducing resuscitation, relieving depression, relieving viscera, smoothing breath, relaxing people, keeping happy mood, preventing common cold, smoothing nasal obstruction, relieving dizziness and headache, stopping dizziness and car and boat sickness, preventing mosquito bites and after bites, relieving swelling and itching, promoting blood circulation, removing blood stasis, dredging channels and collaterals, and effectively treating traumatic injury and sprain are achieved.

Compared with the example 3, the comparative example 7 has no step of S4 fermentation, and the anti-inflammation and mosquito-repellent effects are obviously reduced. The invention adopts fermentation technology, further destroys plant cells, promotes the dissolution of active substances, further improves the extraction rate, and simultaneously contributes to the generation of active aromatic micromolecules in the fermentation process, and the aromatic micromolecules also contribute to improving the effects of preventing cold, smoothing nasal obstruction, relieving dizziness and headache, stopping dizziness and car sickness, preventing mosquito bites and swelling and relieving itching after biting, and the like.

In comparative example 12, compared with example 3, the antibacterial, anti-inflammatory, acaricidal and mosquito repellent effects were reduced without performing step S9. The surface and the interior of the microsphere modified by the polydopamine contain rich polydopamine substances, and the rich hydroxyl, carboxyl, amino and phenyl of the polydopamine are beneficial to forming hydrogen bonds with active substances in essential oil, such as alcohol, ketone, acid, ester and the like, so that the active substances are stably adsorbed, meanwhile, the phenyl is easy to form pi-pi conjugation with aromatic groups in the active substances, such as methyl salicylate and the like, so that the phenyl is stably fixed in the silicon dioxide microsphere, and the carrying capacity of the microsphere on the active substances of the essential oil is improved.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims

1. The preparation method of the multifunctional essential oil composition is characterized by comprising the following steps:

s1, pretreatment of plant materials: microwave drying herba Menthae, eucalyptus globulus Labill, lavender, flos Caryophylli, herba Melissae axillaris, herba Rosmarini officinalis, and herba Agastaches, mixing, and pulverizing to obtain plant material powder; the mass ratio of the mint, the eucalyptus, the lavender, the clove, the melissa officinalis, the rosemary and the patchouli is 1-3:1-2:3-5:2-3:1-2:3-5:5-7;

s2, mixed solvent extraction: adding the plant material powder prepared in the step S1 into a mixed solvent, performing ultrasonic-assisted extraction, filtering, retaining solids, and removing the solvent from the filtrate under reduced pressure to obtain extracted oil;

s3, enzymolysis: adding the solid obtained in the step S2 into water, adding compound enzyme for enzymolysis, and inactivating enzyme to obtain an enzymolysis product; the mass ratio of the solid to the compound enzyme is 10-2;

s5, extracting ionic liquid: adding the fermentation product obtained in the step S4 into the mixed solution of ionic liquid and acetonitrile, and slowly introducing CO ₂ Oscillating, stopping introducing CO ₂ Obtaining a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, adding nano magnetic fluid, oscillating for adsorption, performing magnetic separation, washing, drying and filtering to obtain extraction slurry; the mixed solution of the ionic liquid and the acetonitrile is a mixed solution of the ionic liquid and the acetonitrile according to the mass ratio of 5-7:1-2; the mass ratio of the fermentation product to the nano magnetic fluid to the mixed solution of ionic liquid and acetonitrile is (10-3);

s7, purification: mixing the extracted oil obtained in the step S2 and the extract liquid obtained in the step S6 uniformly, filtering by adopting a small-aperture ceramic membrane, collecting filtrate after small molecules are filtered out, and concentrating by adopting an ultrafiltration membrane to obtain purified volatile oil liquid; the mass ratio of the extraction oil to the extraction liquid is 3-5:2-7;

s8, preparing porous silicon dioxide microspheres: dissolving alkyl orthosilicate in an organic solvent to obtain an oil phase; dissolving a pore-foaming agent and a surfactant in water to obtain a water phase; adding the oil phase into the water phase, emulsifying by an SPG rapid membrane, adjusting the pH value, heating for reaction and solidification, centrifuging, washing and drying to obtain the porous silicon dioxide microspheres; the mass ratio of the alkyl orthosilicate, the pore-forming agent and the surfactant is 10-3;

s9, preparation of the modified porous silicon dioxide microspheres: uniformly dispersing the porous silica microspheres prepared in the step S8 in water, adding dopamine hydrochloride and a catalyst, heating for reaction, centrifuging, washing and drying to obtain modified porous silica microspheres; the mass ratio of the porous silica microspheres to the dopamine hydrochloride to the catalyst is 20-32;

s10, preparing a multifunctional essential oil composition: adding the modified porous silica microspheres obtained in the step S9 into the purified volatile oil liquid obtained in the step S7, stirring, adsorbing and filtering to obtain multifunctional essential oil microspheres, namely the multifunctional essential oil composition; the mass ratio of the modified porous silica microspheres to the purified volatile oil liquid is 10-15.

2. The method according to claim 1, wherein the microwave drying in step S1 is performed at a power of 1000-1500W for 1-2h; the volume ratio of water, ethanol, acetonitrile and petroleum ether in the mixed solvent in the step S2 is 3-5:5-7:1-3:3-5; the solid-liquid ratio of the plant material powder to the mixed solvent is 1:3-5g/mL; the power of the ultrasonic auxiliary extraction is 500-700W, and the time is 30-50min.

3. The preparation method according to claim 1, wherein the complex enzyme in step S3 is selected from at least two of cellulase, pectinase, hemicellulase, amylase and ligninase; the enzymolysis is carried out at 40-60 deg.C for 2-4h.

4. The method according to claim 1, wherein the activated Saccharomyces cerevisiae, bacillus subtilis, or Lactobacillus plantarum is inoculated in step S4The amount of the seed is 3-5%, 2-4% and 0.5-1.5% respectively; the fermentation culture condition is 37-40 ℃, and the time is 24-48h; the activation method comprises respectively inoculating Saccharomyces cerevisiae, bacillus subtilis and Lactobacillus plantarum in Gauss culture medium, culturing at 37-39 deg.C and 50-70r/min for 12-18h to obtain product with bacteria content of 10 ⁸ -10 ⁹ cfu/mL strain seed solution; the ionic liquid in step S5 is selected from 1,3-dimethylimidazole bistrifluoromethylsulfonyl imide salt, 1-ethyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-propyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-butyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-pentyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-hexyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-octyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-decyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-dodecyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-tetradecyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-hexadecyl-3-methylimidazole bistrifluoromethylsulfonyl imide salt, 1-ethyl-1-methylpyrrolidine bistrifluoromethylsulfonyl imide salt, 1-propyl-1-methylpyrrolidine bistrifluoromethylsulfonyl imide salt, 1-butyl-1-methylpyrrolidine bistrifluoromethylsulfonyl imide salt, 1-butyltrifluoromethanesulfonyl-1-butylsulfamoyl imide salt, tri-butyltrifluoromethanesulfonylimide salt, tri-butyltrifluoromethanesulfonimide salt; said slow introduction of CO ₂ Middle CO ₂ The feeding rate of (2) is 0.5-1mL/min, and the feeding time is 30-50min; the nano magnetic fluid is 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ And (4) nano magnetic fluid.

5. The method according to claim 1, wherein the mass ratio of the extraction slurry to the organic solvent in step S6 is 2:3-5; the organic solvent is at least one selected from dichloromethane, trichloromethane, carbon tetrachloride, ethyl acetate, petroleum ether, acetonitrile, methyl acetate and n-hexane; in the step S7, the aperture of the small-aperture ceramic membrane is 2000-3000D, the aperture of the ultrafiltration membrane is 30-100nm, and the relative density of the purified volatile oil liquid is 1.4-1.7.

6. The production method according to claim 1, wherein the alkyl orthosilicate in step S8 is methyl orthosilicate or ethyl orthosilicate; the organic solvent is at least one selected from dichloromethane, trichloromethane, carbon tetrachloride, ethyl acetate, petroleum ether, acetonitrile, methyl acetate and n-hexane; the pore-foaming agent is selected from at least one of polyoxyethylene sorbitan fatty acid ester, polyethylene glycol octyl phenyl ether, cetyl trimethyl ammonium bromide, oxyethylene-oxypropylene triblock copolymer PEO20-PPO70-PEO20 and PEO106-PPO70-PEO 10; the surfactant is at least one of tween-20, tween-40, tween-60, tween-80, sodium dodecyl benzene sulfonate, sodium dodecyl sulfate, sodium dodecyl sulfonate, sodium tetradecyl benzene sulfonate, sodium tetradecyl sulfonate, sodium hexadecyl benzene sulfonate, and sodium octadecyl benzene sulfonate; the pore diameter of the SPG rapid membrane is 5000-7000nm; the pH value is adjusted to 8-9, the temperature of the heating reaction and solidification is 50-70 ℃, and the time is 3-5h; the catalyst in the step S9 is Co containing 3-5wt% ²⁺ The temperature of the heating reaction is 45-55 ℃ and the time is 2-4h.

7. The preparation method according to claim 1, comprising the following steps:

s1, pretreatment of plant materials: drying 1-3 parts by weight of mint, 1-2 parts by weight of eucalyptus, 3-5 parts by weight of lavender, 2-3 parts by weight of clove, 1-2 parts by weight of melissa officinalis, 3-5 parts by weight of rosemary and 5-7 parts by weight of patchouli by 1000-1500W microwave for 1-2h, mixing, crushing, and sieving with a 60-100 mesh sieve to obtain plant material powder;

the activation method comprises respectively inoculating Saccharomyces cerevisiae, bacillus subtilis and Lactobacillus plantarum in Gao's medium, culturing at 37-39 deg.C and 50-70r/min for 12-18h to obtain extract with bacteria content of 10 ⁸ -10 ⁹ cfu/mL of strain seed solution;

s5, extracting ionic liquid: adding 10 parts by weight of the fermentation product obtained in the step S4 into 15-20 parts by weight of the ionic liquid-acetonitrile mixed solution, and introducing CO ₂ Introducing CO at a rate of 0.5-1mL/min for 30-50min, oscillating, and stopping introducing CO ₂ Obtaining a high-extraction ionic liquid-acetonitrile mixed solution, standing for layering, separating liquid, removing a water layer, and adding 2-3 parts by weight of 1-hydroxyethyl-3-methylimidazolyl Fe ₃ O ₄ Nano magnetic fluid, oscillating adsorption, magnetic separation, washing, drying and filtering to obtain extraction slurry;

the ionic liquid-acetonitrile mixed solution is a mixed solution of ionic liquid and acetonitrile according to the mass ratio of 5-7:1-2;

s7, purification: 3-5 parts by weight of the extracted oil obtained in the step S2 and 2-7 parts by weight of the extract liquid obtained in the step S6 are uniformly mixed, the mixture is filtered by adopting a small-pore-diameter ceramic membrane with the pore diameter of 2000-3000D, the filtrate obtained after small molecule filtration is collected, and the filtrate is concentrated by adopting an ultrafiltration membrane with the pore diameter of 30-100nm to obtain purified volatile oil liquid with the relative density of 1.4-1.7;

s8, preparing porous silicon dioxide microspheres: dissolving 10 parts by weight of methyl orthosilicate or ethyl orthosilicate in 50 parts by weight of organic solvent to obtain an oil phase; dissolving 2-3 parts by weight of pore-foaming agent and 0.5-1 part by weight of surfactant in 100 parts by weight of water to obtain a water phase; adding the oil phase into the water phase, emulsifying with SPG rapid membrane with aperture of 5000-7000nm, adjusting pH to 8-9, heating to 50-70 deg.C, reacting and curing for 3-5h, centrifuging, washing, and drying to obtain porous silica microsphere;

8. A multifunctional essential oil composition prepared by the preparation method according to any one of claims 1 to 7.

9. The use of the multifunctional essential oil composition of claim 8 in the preparation of related products for refreshing brain, inducing resuscitation, resolving stagnation, preventing common cold, smoothing nasal obstruction, relieving dizziness and headache, stopping dizziness and motion sickness, preventing mosquito bites and detumescence and relieving itching after biting.