CN111671812A - Licorice extract and essential oil composite nanoemulsion preparation and preparation method thereof - Google Patents
Licorice extract and essential oil composite nanoemulsion preparation and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/484—Glycyrrhiza (licorice)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/534—Mentha (mint)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/75—Rutaceae (Rue family)
- A61K36/752—Citrus, e.g. lime, orange or lemon
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
- A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
Abstract
The invention relates to a liquorice extract and essential oil composite nanoemulsion preparation and a preparation method thereof, wherein the method comprises the following steps: s1, mixing the crude licorice extract with essential oil, stirring and heating for 1-6h at 20-35 ℃ and at the rotation speed of 500-800rpm to obtain cream yellow oily liquid as an oil phase; s2, weighing the surfactant Tween 20 and the deionized water according to the mass ratio of 1:10, and then fully mixing at room temperature and the rotation speed of 500-800rpm to obtain a clear aqueous solution which is an external aqueous phase; s3, dropwise dispersing the oil phase into the water phase until the oil phase is completely dispersed, and then obtaining a yellow opaque emulsion which is a crude licorice emulsion by high shear; s4, homogenizing the crude emulsion under high pressure to obtain semitransparent emulsion which is licorice and essential oil-in-water type nano emulsion.
Description
Technical Field
The invention relates to a nanoemulsion preparation, in particular to a liquorice extract and essential oil composite nanoemulsion preparation and a preparation method thereof, and belongs to the technical field of functional food additives.
Background
Caries is a common and frequently encountered disease of human oral cavity, and the world health organization has listed caries as one of three major non-infectious diseases for human key prevention and treatment. Streptococcus mutans is one of the resident bacteria in the oral cavity and is also the main pathogenic bacteria of caries. Fluoride has been used for preventing and treating caries for more than 60 years, is the most common preparation for preventing caries at present, and the effectiveness of preventing caries is well recognized and widely accepted. On the other hand, the use of fluoride also shows increasingly significant limitations, such as long-term stability of the fluorine concentration in fluorine-containing toothpastes; after the fluoride is used for a long time, the streptococcus mutans generates a fluorine-resistant strain, and the genome of bacteria is changed, so that the caries prevention effect of the fluoride is reduced; acute and chronic poisoning of the body caused by improper use of fluoride greatly affects the application and popularization of fluoride in caries prevention, and brings great hidden danger to the health of people. In recent years, researches show that certain traditional Chinese medicines such as liquorice and the like also have the capacity of inhibiting the growth and acid production of streptococcus mutans and have the anti-caries effect similar to chemical preparations such as chlorhexidine and sodium fluoride. Liquorice, honeysuckle, propolis, platycodon grandiflorum and radix puerariae are considered to have antibacterial and anti-inflammatory effects in Chinese traditional medicine research, but reports of application to treatment of oral diseases are not found at present.
The naturally extracted plant essential oil also has the effects of resisting bacteria, inflammation and cancer, and the like, and because the plant active molecules have strong synergistic interaction, after two or more plant extracts are mixed according to a certain proportion, the biological effects of the respective active molecules can be fully exerted to the utmost extent, and the effect which cannot be achieved by the plant extract with a single component is achieved, so the liquorice extract and the essential oil are reasonably and effectively used in a composite way, and the unexpected oral care effect can be achieved.
The chemical components of licorice are numerous, and about 450 compounds have been isolated and reported from glycyrrhiza plants so far, and are mainly divided into triterpenoid saponins, flavonoid aglycones and flavone glycoside compounds. Wherein the aglycone of triterpene saponin is mainly oleanane type triterpene, while flavonoid aglycone compounds are various, and mainly comprise flavone, flavonol, flavanone, isoflavone, dihydroisoflavone, isoflavane, pterocarpan, chalcone and coumarin. Most of the flavonoids are water-insoluble. At present, common methods for improving the water solubility of liquorice include a microencapsulation method, a self-emulsifying system, a liposome, an inclusion compound, an emulsion and the like. The oil-in-water nano dispersion system has small and uniform particle size and is beneficial to absorption and administration of cells. The following advantages are also provided: (1) the safety of the pharmaceutical preparation is improved; (2) the solubility and the dissolution rate of the drug are improved; (3) the stability of the pharmaceutical preparation is improved; (4) the biological adhesiveness of the medicine is improved; (5) realizing targeted drug delivery.
Disclosure of Invention
The invention provides a liquorice extract and essential oil composite nanoemulsion preparation and a preparation method thereof aiming at the problems in the prior art, the technical scheme provides a liquorice crude extract and plant essential oil composite nanoemulsion preparation and a preparation method thereof, and the essential oil comprises: origanum essential oil, mint essential oil, basil essential oil, lemon essential oil, orange peel essential oil and bergamot essential oil, aiming at increasing the solubility of liquorice and solving the problem of dental caries.
In order to achieve the purpose, the technical scheme of the invention is as follows, the licorice extract and essential oil composite nanoemulsion preparation and the preparation method thereof comprise the following steps:
s1, mixing the crude licorice extract with essential oil, stirring and heating for 1-2h at 20-35 ℃ and 800rpm to obtain cream yellow oily liquid as an oil phase;
s2, weighing the surfactant Tween 20 and the deionized water according to the mass ratio of 1:10, and then fully mixing at room temperature and the rotation speed of 500-800rpm to obtain a clear aqueous solution which is an external aqueous phase;
s3, dropwise dispersing the oil phase into the water phase until the oil phase is completely dispersed, and then obtaining a yellow opaque emulsion which is a crude licorice emulsion by high shear;
s4, homogenizing the crude emulsion under high pressure to obtain semitransparent emulsion which is licorice and essential oil-in-water type nano emulsion.
As a modification of the present invention, in this step S1, the mass ratio of licorice and essential oil is 1: 1. Thus, the using amount of the essential oil is small, and the finally prepared nano-emulsion is more stable.
As a modification of the present invention, in step S3, the mass ratio of the oil phase to the external water phase is 1:10, and the conditions for preparing the crude emulsion under high shear are 10000-12000rpm for 5-10 min. The good coarse emulsion can be obtained within proper shearing time, the subsequent preparation of the nano emulsion is facilitated, and the demulsification phenomenon can be caused by too long shearing time.
As a modification of the invention, in the step S4, the oil-in-water type nano emulsion is prepared under high pressure and homogeneous conditions of 30-50MPa for 15-20 times.
As an improvement of the present invention, the essential oil comprises: one or two of oregano essential oil, mint essential oil, basil essential oil, lemon essential oil, orange peel essential oil and bergamot essential oil are mixed in a mass ratio of (1-3) to 1. Aims to increase the solubility of the liquorice and solve the problem of dental caries.
The nanoemulsion preparation prepared by the preparation method is an oil-in-water type nanoemulsion formed by an oil phase (liquorice and essential oil), a surfactant Tween 20 and water-phase deionized water; in the oil phase, the mass ratio of the liquorice to the essential oil is 2: 1, the mass ratio of the internal oil phase of the emulsion particles to the external water phase of the emulsion is 1: 10.
Compared with the prior art, the invention has the advantages that (1) the prepared emulsion is free of other auxiliary materials except the emulsifier, and all the raw materials are edible, so that the biological safety of the licorice root nanoemulsion preparation is ensured; (2) the preparation condition is mild, no high temperature exists, the loss of the liquorice and the essential oil in the preparation process is reduced, and the embedding rate of the liquorice is improved; (3) the prepared emulsion is yellow and semitransparent, and the average particle size of the emulsion is Z-average: 160 +/-5 nm, the potential Zeta is-32.6 +/-0.5 mV, and the dispersion index PDI is 0.18, which shows that the emulsion has good stability and dispersibility and can improve the water solubility of the liquorice. (4) The liquorice and the essential oil are compounded together, so that the synergistic effect can be exerted, and the antibacterial effect is greatly improved; (5) the volatile and inherent fragrance of the essential oil can not only treat caries, but also refresh breath.
Drawings
FIG. 1: the invention discloses a preparation flow chart of a liquorice crude extract and an essential oil nanoemulsion.
The specific implementation mode is as follows:
for the purpose of enhancing an understanding of the present invention, the present embodiment will be described in detail below with reference to the accompanying drawings. However, the present invention is not limited to the following examples.
Example 1: referring to fig. 1, a specific preparation method of a licorice crude extract and essential oil composite nanoemulsion comprises the following steps:
s1, accurately weighing 0.5g of licorice crude extract and 0.5g of essential oil, placing the licorice crude extract and the essential oil in a centrifuge tube, and vortexing for 5-10min to fully mix the licorice crude extract and the essential oil to obtain cream yellow oily liquid which is an oil phase;
s2, accurately weighing 5g of surfactant Tween 20 and 50g of deionized water, placing the weighed materials into an 80ml beaker, placing the beaker on a magnetic stirrer for stirring, setting the temperature at 25 ℃, regulating the rotating speed to 500rpm, and stirring for 30min to obtain a clear aqueous solution which is an external water phase;
s3, slowly adding the oil phase dropwise, primarily dispersing the oil phase into the external water phase until the components are completely dispersed, and preparing the suspension by high shear, wherein the shearing conditions are as follows: the rotation speed is 12000rpm, the time is 5min, and yellow opaque emulsion is obtained, which is licorice crude emulsion.
S4, homogenizing the crude licorice emulsion under high pressure, wherein the homogenizing conditions are as follows: the pressure is 30MPa, and the number of times of homogenizing is 15. During each homogenization, the emulsion is cooled by circulating water to prevent the emulsion from influencing the emulsified particle size due to overhigh temperature, and finally, the yellow emulsion which is the liquorice and essential oil-in-water type nano emulsion is obtained.
Example 2: preparation of liquorice crude extract and essential oil composite nanoemulsion
S1, accurately weighing 0.5g of liquorice crude extract and 0.75g of essential oil, placing the liquorice crude extract and the essential oil in a centrifugal tube, and vortexing for 5-10min to fully mix the liquorice crude extract and the essential oil to obtain creamy yellow oily liquid serving as an oil phase;
s2, accurately weighing 5g of surfactant Tween 20 and 50g of deionized water, placing the weighed materials into an 80ml beaker, placing the beaker on a magnetic stirrer for stirring, setting the temperature at 25 ℃, regulating the rotating speed to 500rpm, and stirring for 30min to obtain a clear aqueous solution which is an external water phase;
s3, slowly adding the oil phase dropwise, primarily dispersing the oil phase into the external water phase until the components are completely dispersed, and preparing the suspension by high shear, wherein the shearing conditions are as follows: the rotation speed is 12000rpm, the time is 5min, and yellow opaque emulsion is obtained, which is licorice crude emulsion.
S4, homogenizing the crude licorice emulsion under high pressure, wherein the homogenizing conditions are as follows: the pressure is 30MPa, and the number of times of homogenizing is 15. During each homogenization, the emulsion is cooled by circulating water to prevent the emulsion from influencing the emulsified particle size due to overhigh temperature, and finally, the yellow emulsion which is the liquorice and essential oil-in-water type nano emulsion is obtained.
Example 3: preparation of liquorice crude extract and essential oil composite nanoemulsion
S1, accurately weighing 0.5g of liquorice crude extract and 1.0g of essential oil, placing the liquorice crude extract and the essential oil in a centrifugal tube, and vortexing for 5-10min to fully mix the liquorice crude extract and the essential oil to obtain creamy yellow oily liquid serving as an oil phase;
s2, accurately weighing 5g of surfactant Tween 20 and 50g of deionized water, placing the weighed materials into an 80ml beaker, placing the beaker on a magnetic stirrer for stirring, setting the temperature at 25 ℃, regulating the rotating speed to 500rpm, and stirring for 30min to obtain a clear aqueous solution which is an external water phase;
s3, slowly adding the oil phase dropwise, primarily dispersing the oil phase into the external water phase until the components are completely dispersed, and preparing the suspension by high shear, wherein the shearing conditions are as follows: the rotation speed is 12000rpm, the time is 5min, and yellow opaque emulsion is obtained, which is licorice crude emulsion.
S4, homogenizing the crude licorice emulsion under high pressure, wherein the homogenizing conditions are as follows: the pressure is 30MPa, and the number of times of homogenizing is 15. During each homogenization, the emulsion is cooled by circulating water to prevent the emulsion from influencing the emulsified particle size due to overhigh temperature, and finally, the yellow emulsion which is the liquorice and essential oil-in-water type nano emulsion is obtained.
Example 4: a method for preparing compound nanoemulsion of Glycyrrhrizae radix crude extract and essential oil is provided.
S1, accurately weighing 0.5g of liquorice crude extract and 0.25g of essential oil, placing the liquorice crude extract and the essential oil in a centrifugal tube, and vortexing for 5-10min to fully mix the liquorice crude extract and the essential oil to obtain creamy yellow oily liquid serving as an oil phase;
s2, accurately weighing 5g of surfactant Tween 20 and 50g of deionized water, placing the weighed materials into an 80ml beaker, placing the beaker on a magnetic stirrer for stirring, setting the temperature at 25 ℃, regulating the rotating speed to 500rpm, and stirring for 30min to obtain a clear aqueous solution which is an external water phase;
s3, slowly adding the oil phase dropwise, primarily dispersing the oil phase into the external water phase until the components are completely dispersed, and preparing the suspension by high shear, wherein the shearing conditions are as follows: the rotation speed is 12000rpm, the time is 5min, and yellow opaque emulsion is obtained, which is licorice crude emulsion.
S4, homogenizing the crude licorice emulsion under high pressure, wherein the homogenizing conditions are as follows: the pressure is 30MPa, and the number of times of homogenizing is 15. During each homogenization, the emulsion is cooled by circulating water to prevent the emulsion from influencing the emulsified particle size due to overhigh temperature, and finally, the yellow emulsion which is the liquorice and essential oil-in-water type nano emulsion is obtained.
Embodiment 5. preparation of licorice crude extract and essential oil composite nanoemulsion
S1, accurately weighing 0.5g of licorice crude extract and 0.5g of essential oil, placing the licorice crude extract and the essential oil in a centrifuge tube, and vortexing for 5-10min to fully mix the licorice crude extract and the essential oil to obtain cream yellow oily liquid which is an oil phase;
s2, accurately weighing 4g of surfactant Tween 20 and 50g of deionized water, placing the weighed materials into an 80ml beaker, placing the beaker on a magnetic stirrer to stir, setting the temperature at 25 ℃, regulating the rotating speed to 500rpm, and stirring for 30min to obtain a clear aqueous solution which is an external water phase;
s3, slowly adding the oil phase dropwise, primarily dispersing the oil phase into the external water phase until the components are completely dispersed, and preparing the suspension by high shear, wherein the shearing conditions are as follows: the rotation speed is 12000rpm, the time is 5min, and yellow opaque emulsion is obtained, which is licorice crude emulsion.
S4, homogenizing the crude licorice emulsion under high pressure, wherein the homogenizing conditions are as follows: the pressure is 30MPa, and the number of times of homogenizing is 15. During each homogenization, the emulsion is cooled by circulating water to prevent the emulsion from influencing the emulsified particle size due to overhigh temperature, and finally, the yellow emulsion which is the liquorice and essential oil-in-water type nano emulsion is obtained.
Embodiment 6 preparation of liquorice crude extract and essential oil composite nanoemulsion
S1, accurately weighing 0.5g of licorice crude extract and 0.5g of essential oil, placing the licorice crude extract and the essential oil in a centrifuge tube, and vortexing for 5-10min to fully mix the licorice crude extract and the essential oil to obtain cream yellow oily liquid which is an oil phase;
s2, accurately weighing 4.3g of surfactant Tween 20 and 50g of deionized water, placing the weighed materials into an 80ml beaker, placing the beaker on a magnetic stirrer to stir, setting the temperature at 25 ℃, adjusting the rotating speed to 500rpm, and stirring for 30min to obtain a clear aqueous solution which is an external water phase;
s3, slowly adding the oil phase dropwise, primarily dispersing the oil phase into the external water phase until the components are completely dispersed, and preparing the suspension by high shear, wherein the shearing conditions are as follows: the rotation speed is 12000rpm, the time is 5min, and yellow opaque emulsion is obtained, which is licorice crude emulsion.
S4, homogenizing the crude licorice emulsion under high pressure, wherein the homogenizing conditions are as follows: the pressure is 30MPa, and the number of times of homogenizing is 15. During each homogenization, the emulsion is cooled by circulating water to prevent the emulsion from influencing the emulsified particle size due to overhigh temperature, and finally, the yellow emulsion which is the liquorice and essential oil-in-water type nano emulsion is obtained.
Embodiment 7. preparation of liquorice crude extract and essential oil composite nanoemulsion
S1, accurately weighing 0.5g of licorice crude extract and 0.5g of essential oil, placing the licorice crude extract and the essential oil in a centrifuge tube, and vortexing for 5-10min to fully mix the licorice crude extract and the essential oil to obtain cream yellow oily liquid which is an oil phase;
s2, accurately weighing 4.5g of surfactant Tween 20 and 50g of deionized water, placing the weighed materials into an 80ml beaker, placing the beaker on a magnetic stirrer to stir, setting the temperature at 25 ℃, adjusting the rotating speed to 500rpm, and stirring for 30min to obtain a clear aqueous solution which is an external water phase;
s3, slowly adding the oil phase dropwise, primarily dispersing the oil phase into the external water phase until the components are completely dispersed, and preparing the suspension by high shear, wherein the shearing conditions are as follows: the rotation speed is 12000rpm, the time is 5min, and yellow opaque emulsion is obtained, which is licorice crude emulsion.
S4, homogenizing the crude licorice emulsion under high pressure, wherein the homogenizing conditions are as follows: the pressure is 30MPa, and the number of times of homogenizing is 15. During each homogenization, the emulsion is cooled by circulating water to prevent the emulsion from influencing the emulsified particle size due to overhigh temperature, and finally, the yellow emulsion which is the liquorice and essential oil-in-water type nano emulsion is obtained.
Example 8 the prepared nanoemulsion was finally stored in an environment at 4 ℃ and the number of days of storage was observed.
The days of storage of the glycyrrhiza crude extract and essential oil composite nanoemulsion prepared in examples 1 to 7 are shown in the following table:
| example of the implementation | Days of storage | Appearance characteristics |
| Example 1 | More than 6 months | The color and luster are uniform after 6 months without layering |
| Example 2 | For 5 months | Precipitate appeared after standing for 5 months |
| Embodiment 3 | 3 months old | Precipitate appeared after standing for 3 months |
| Example 4 | 7 days | Precipitate appeared after standing for 7 days |
| Example 5 | 3 days | Precipitate appeared after standing for 3 days |
| Example 6 | 5 days | Precipitate appeared after standing for 5 days |
| Example 7 | 5 days | Precipitate appeared after standing for 5 days |
From the above table, it can be seen that: the liquorice crude extract and essential oil composite nanoemulsion prepared in the embodiment 1 has the longest storage time at 4 ℃, and no obvious layering phenomenon is found in 6 months, which indicates that the prepared nanoemulsion has good stability. Thus, example 1 was used as the preparation formulation and preparation conditions of the present invention.
Example 9: after the nano-emulsions prepared by the different formulas of the cases 1 to 7 are stored at low temperature for one week, the nano-emulsions are respectively centrifuged for 30 minutes at 4 ℃ and 3000r, the precipitates are redissolved in ethanol, and after being filtered by a 0.45 mu m microporous filtering membrane, the content of the liquorice in the precipitates is detected by adopting high performance liquid chromatography, and the solubility of the liquorice in the emulsions is calculated.
The solubility of licorice, which is analytically measured for the licorice extracts and essential oil composite nano-emulsions prepared in examples 1-7, is shown in the following table:
| example of the implementation | Solubility of Glycyrrhiza |
| Example 1 | 94％ |
| Example 2 | 96％ |
| Embodiment 3 | 98.2％ |
| Example 4 | 60％ |
| Example 5 | 73％ |
| Example 6 | 85％ |
| Example 7 | 90％ |
From the above table, it can be seen that: after the prepared licorice extract and the essential oil nanoemulsion are placed at a low temperature for one week, the solubility of the licorice extract prepared in embodiment 1, embodiment 2 and embodiment 3 and licorice in the essential oil nanoemulsion reaches more than 90%, and it can be seen that the water solubility of the licorice can be greatly improved by the licorice crude extract and the essential oil nanoemulsion prepared by the invention.
Example 10: streptococcus mutans is the leading causative bacterium of dental caries. Streptococcus mutans is easy to produce acid and has strong acid resistance, can continue to live and produce acid when the pH is 4.5, and easily dissolves calcium, thus causing local hard tissue of teeth to fall off and dental caries. The streptococcus mutans takes sucrose as a substrate to synthesize water-insoluble and strongly adhesive polysaccharide which is firmly adhered to the tooth surface to accelerate the formation of bacterial plaque and cause caries. The detection rate of the porphyromonas gingivalis in patients with chronic periodontitis is obviously higher than that of patients with periodontal health, and the detection rate tends to gradually increase along with the increase of the depth of periodontal probing. Porphyromonas gingivalis causes inflammation of gingiva, also generates resistance to common antibiotics, and squeezes out and replaces bacteria beneficial to teeth, and once the porphyromonas gingivalis is out of control, teeth are loosened and fall off finally. Therefore, control of streptococcus mutans and porphyromonas gingivalis is critical for reducing caries and preventing periodontal disease.
The oral pathogenic bacteria are Streptococcus mutans (Streptococcus mutans) CMCC32400 or Porphyromonas gingivalis (Porphyromonas gingivalis) ATCC 33277.
When the test strain in the following examples is Streptococcus mutans (Streptococcus mutans) CMCC32400, the test method is: the test strains were incubated at 37 ℃ for 24h using Brain Heart Infusion Agar (BHIA) slant medium in this example. Then, the cells were diluted with PBS until OD600 became 0.13 to prepare a working solution of the cell suspension. Then adding the emulsion, the licorice extract or the essential oil into a 96-well plate, and measuring the minimum inhibitory concentration. Culturing at 37 deg.C for 24 hr, and adding 0.0015% resazurin color developing solution.
The Minimum Inhibitory Concentrations (MIC) of the three substances against S.mutans are given in the following table:
| additive substance | Minimum inhibitory concentration (μ g/mL) |
| Glycyrrhiza extract | 250 |
| Essential oil | 100 |
| Composite nanoemulsion of liquorice crude extract and essential oil | 15.6 |
The above table shows that the licorice crude extract and the essential oil composite nanoemulsion prepared by the invention have a synergistic effect in the process of inhibiting streptococcus mutans, and compared with a single licorice extract or a single essential oil, the bacteriostatic activity of the licorice crude extract and the essential oil composite nanoemulsion is greatly improved.
It should be noted that the above-mentioned embodiments are not intended to limit the scope of the present invention, and all equivalent modifications and substitutions based on the above-mentioned technical solutions are within the scope of the present invention as defined in the claims.
Claims (7)
1. The liquorice extract and essential oil composite nanoemulsion preparation and the preparation method thereof are characterized by comprising the following steps:
s1, mixing the crude extract of the liquorice with the essential oil, stirring and heating for 1-2h at the temperature of 20-35 ℃ and the rotation speed of 500-800rpm to obtain cream yellow oily liquid which is an oil phase;
s2, weighing the surfactant Tween 20 and the deionized water according to the mass ratio of 1:10, and then fully mixing at room temperature and the rotation speed of 500-800rpm to obtain a clear aqueous solution which is an external aqueous phase;
s3, dropwise dispersing the oil phase into the water phase until the oil phase is completely dispersed, and then obtaining a yellow opaque emulsion which is a crude licorice emulsion by high shear;
s4, homogenizing the crude emulsion under high pressure to obtain semitransparent emulsion which is licorice and essential oil-in-water type nano emulsion.
2. The licorice extract and essential oil composite nanoemulsion formulation and the preparation method thereof according to claim 1, wherein in step S1, the mass ratio of licorice to essential oil is 1: 1.
3. The licorice extract and essential oil composite nanoemulsion formulation and preparation method of claim 2, wherein in step S3, the mass ratio of the oil phase to the external water phase is 1:10, and the conditions for preparing the crude emulsion under high shear are 10000-12000rpm for 5-10 min.
4. The licorice extract and essential oil composite nanoemulsion formulation and the preparation method thereof according to claim 3, wherein in step S4, the oil-in-water nanoemulsion formulation is prepared under the high-pressure homogenization condition of 30-50MPa for 15-20 times.
5. The licorice extract and essential oil composite nanoemulsion formulation and preparation method of claims 3 or 4, wherein the essential oil comprises: one or two of oregano essential oil, mint essential oil, basil essential oil, lemon essential oil, orange peel essential oil and bergamot essential oil are mixed in a mass ratio of (1-3) to 1.
6. The nanoemulsion formulation prepared by the preparation method according to any one of claims 1 to 6, wherein the nanoemulsion formulation is an oil-in-water nanoemulsion formed by an oil phase (licorice extract and essential oil), a surfactant Tween 20, and an aqueous phase deionized water; in the oil phase, the mass ratio of the liquorice to the essential oil is 2: 1, the mass ratio of the internal oil phase of the emulsion particles to the external water phase of the emulsion is 1: 10.
7. The nanometer emulsion is used in oral cavity medicine or oral cavity nursing product for inhibiting pathogenic bacteria in oral cavity, treating dental caries, reducing dental plaque and gingivitis, preventing halitosis, etc.
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|---|---|---|---|---|
| CN101147724A (en) * | 2006-12-04 | 2008-03-26 | 好维股份有限公司 | Medicine or sanitary product containing licorice flavone extraction for oral cavity |
| CN104606085A (en) * | 2013-11-22 | 2015-05-13 | 中国人民解放军第三军医大学 | Novel efficient antimicrobial chlorhexidine acetate nanoemulsion gargle and preparation method thereof |
| CN111135142A (en) * | 2020-01-16 | 2020-05-12 | 兰州大学 | Isoliquiritigenin nanoemulsion and preparation method thereof |
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2020
- 2020-07-01 CN CN202010625044.9A patent/CN111671812A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101147724A (en) * | 2006-12-04 | 2008-03-26 | 好维股份有限公司 | Medicine or sanitary product containing licorice flavone extraction for oral cavity |
| CN104606085A (en) * | 2013-11-22 | 2015-05-13 | 中国人民解放军第三军医大学 | Novel efficient antimicrobial chlorhexidine acetate nanoemulsion gargle and preparation method thereof |
| CN111135142A (en) * | 2020-01-16 | 2020-05-12 | 兰州大学 | Isoliquiritigenin nanoemulsion and preparation method thereof |
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| 孟胜男: "纳米乳与亚微乳", 《药剂学》 * |
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