Chinese herbal medicine composition antibacterial nanoemulsion and preparation method thereof

Abstract

The invention relates to the technical field of biological medicine, and discloses a Chinese herbal medicine composition antibacterial nanoemulsion and a preparation method thereof, the Chinese herbal medicine composition antibacterial nanoemulsion comprises the following components by weight: 20-40 parts of a Chinese herbal medicine extract; 25 to 40 parts of a solvent; 5 to 12 parts of an oil phase carrier; 8 to 15 parts of an emulsifier; 3-8 parts of a co-emulsifier; 1-3 parts of a co-stabilizer; the Chinese herbal medicine extract is prepared from honeysuckle flowers, radix scutellariae, radix sophorae flavescentis and cortex phellodendri, and the weight ratio of the honeysuckle flowers to the radix scutellariae to the radix sophorae flavescentis to the cortex phellodendri is (3-6): (2-4): (1-2): (0.5-1). According to the Chinese herbal medicine composition antibacterial nanoemulsion, Chinese herbal medicines such as honeysuckle, radix scutellariae, radix sophorae flavescentis and cortex phellodendri are used as raw materials, natural antibacterial activity is achieved through the synergistic effect of the Chinese herbal medicines, pathogenic bacteria are not prone to generating drug resistance, use is safe and environmentally friendly, drug residues are avoided, and the problems of drug resistance and drug residues existing in chemical antibiotics are effectively solved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to a Chinese herbal medicine composition antibacterial nanoemulsion and a preparation method thereof. BACKGROUND

[0002] Aquaculture is an important source of protein worldwide, but bacterial diseases seriously affect the benefits of aquaculture, especially the characteristic fish of Guizhou Megalobrama amblycephala, which is more prone to bacterial infection in high-density culture mode. Traditional antibiotics have been restricted in many countries due to the problems of increasing drug resistance, drug residues, etc. Nanoemulsion technology can improve the solubility of hydrophobic drugs, enhance intestinal absorption, and achieve sustained release or targeted delivery, so it has broad prospects in the field of aquaculture antibacterial.

[0003] In the process of aquaculture, the outbreak of bacterial diseases often brings huge economic losses to the aquaculture industry. Traditional prevention and control methods rely on chemical antibiotics, but long-term and large-scale use of chemical antibiotics can easily lead to drug resistance of pathogenic bacteria, and drug residues can affect the quality of aquatic products and human health, and also cause potential harm to the ecological environment. SUMMARY

[0004] In view of the deficiencies of the prior art, the present application provides a Chinese herbal medicine composition antibacterial nanoemulsion and a preparation method thereof, which solves the problems of drug resistance, drug residues and destruction of the ecological environment caused by chemical antibiotics.

[0005] To achieve the above purpose, the present application realizes the following technical scheme: a Chinese herbal medicine composition antibacterial nanoemulsion and a preparation method thereof, comprising the following components by weight: Chinese herbal medicine extract 20-40 parts; Solvent 25-40 parts; Oil phase carrier 5-12 parts; Emulsifier 8-15 parts; Co-emulsifier 3-8 parts; Co-stabilizer 1-3 parts; The balance is the water phase.

[0006] Preferably, the Chinese herbal medicine extract is prepared from honeysuckle, radix scutellariae, sophora and phellodendron, and the weight ratio of honeysuckle, radix scutellariae, sophora and phellodendron is (3-6):(2-4):(1-2):(0.5-1).

[0007] Preferably, the solvent is one or a combination of more than one of ethanol, propylene glycol or glycerol; The oil phase carrier is one or a combination of more than one of olive oil, tea seed oil or soybean oil; The co-stabilizer is one or a combination of more than one of xanthan gum, gum arabic or sodium carboxymethyl cellulose.

[0008] Preferably, the emulsifier is one or more of a combination of Tween-80, Span-80 or polyoxyethylene castor oil; The co-emulsifier is any one of n-butanol, ethylene glycol monomethyl ether or propylene glycol methyl ether.

[0009] Preferably, the nanoemulsion particle size is 80-200nm, and the polydispersity index is ≤0.25.

[0010] A preparation method of a Chinese herbal medicine composition antibacterial nanoemulsion, comprising the following steps: Step one, grind honeysuckle, radix scrophulariae, sophora flavescens and cortex phellodendri to 80-100 mesh, respectively, for standby; Step two, after the raw material powdering in step one is completed, the Chinese herbal medicine raw material mixture is obtained by mixing, and then the solvent is added, the amount of solvent added is 5-8 times the total weight of the raw material, and the reflux extraction method is used to extract at 65-75℃ for 2.5-3.5 hours to obtain the extract; Step three, filter the obtained extract, and concentrate the filtrate under reduced pressure to a relative density of 1.10-1.15 at 40-50℃ to obtain a Chinese herbal medicine extract; Step four, after obtaining the Chinese herbal medicine extract, mix the Chinese herbal medicine extract, solvent, emulsifier, co-emulsifier and oil phase carrier, stir at a speed of 3000-4000r / min for 10-15 minutes to obtain a mixed liquid A; Step five, add a co-stabilizer to the mixed liquid A, and continue to stir at a speed of 3000-4000r / min for 5-8 minutes to obtain a mixed liquid B; Step six, slowly add the mixed liquid B to the water phase at 50-60℃, the amount of water phase added is 3-5 times the total weight of the mixed liquid B, and stir at a speed of 2000-3000r / min for 20-30 minutes while adding, and then cool to room temperature to obtain a Chinese herbal medicine composition antibacterial nanoemulsion.

[0011] Preferably, in step two, the solvent is a combination of ethanol and propylene glycol, and the weight ratio of ethanol to propylene glycol is 3:1.

[0012] Preferably, in step four, the emulsifier is a combination of Tween-80 and Span-80, and the weight ratio of Tween-80 to Span-80 is 2:1, and the co-emulsifier is n-butanol.

[0013] Preferably, the water phase is any one of distilled water or phosphate buffer.

[0014] Preferably, in step six, the nanoemulsion after cooling to room temperature is stored at 1-4℃.

[0015] The application provides a Chinese herbal medicine composition antibacterial nanoemulsion and a preparation method thereof. 1、The Chinese herbal medicine composition antibacterial nanoemulsion is prepared from honeysuckle, radix scutellariae, radix sophorae flavescentis and radix phellodendri as raw materials, and natural antibacterial activity is exerted through the synergistic effect of the Chinese herbal medicines, so that pathogenic bacteria are not prone to drug resistance, the Chinese herbal medicine composition antibacterial nanoemulsion is safe and environmentally friendly, and no drug residues are generated, thus effectively solving the problems of drug resistance and drug residues of chemical antibiotics and guaranteeing the quality and safety of aquatic products.

[0016] 2、The Chinese herbal medicine extract is prepared into nanoemulsion, the characteristics of the nanoemulsion are utilized, the effective components of the Chinese herbal medicine are fully reserved and uniformly dispersed, the contact efficiency of the antibacterial components and pathogenic bacteria is improved, the water solubility and bioavailability of the Chinese herbal medicine extract are improved, the antibacterial effect is enhanced, and the pathogenic bacteria in aquaculture can be more effectively inhibited.

[0017] 3、The emulsifier, the co-emulsifier and the oil phase carrier are combined, a stable colloidal dispersion system of the nanoemulsion is realized, and the problems of easy delamination and emulsion breaking of the product during transportation and use are solved. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 The application further provides a step flow chart of the preparation method of the Chinese herbal medicine composition antibacterial nanoemulsion. DETAILED DESCRIPTION

[0019] The technical solutions of the application will be described clearly and completely in combination with the drawings of the application. Apparently, the described embodiments are only some of the embodiments of the application, but not all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the application.

[0020] The application will be further described in combination with examples and comparative examples.

[0021] The honeysuckle is selected from Guizhou Bailin Pharmaceutical Co., Ltd. and has a brand BL-JYH-01; The radix scutellariae is selected from Chengde Jingfukang Pharmaceutical Co., Ltd. and has a brand JF-HQ-GMP; The radix sophorae flavescentis is selected from Lanzhou Foci Pharmaceutical Co., Ltd. and has a brand FC-KS-202; The radix phellodendri is selected from Sichuan Xinhua Chinese Medicine Co., Ltd. and has a brand XHH-HB-60; The Tween-80 is selected from Klora and has a brand Crill 10; The Span-80 is selected from Huoda and has a brand Span 80; The polyoxyethylene castor oil is selected from BASF and has a brand Cremophor EL Preparation Example I: Preparation of Chinese herbal medicine extract: Honeysuckle, Huangqi, Kuhsiang and Huangbai are mixed in a weight ratio of 3:2:1:0.5, respectively crushed to 80 mesh, 6 times the weight of 75% ethanol solution is added, refluxed at 70℃ for 3 hours, filtered and concentrated under reduced pressure to a relative density of 1.12 extract, the Chinese herbal medicine extract is obtained.

[0022] Preparation Example Two: Preparation of Chinese herbal medicine extract: Honeysuckle, Huangqi, Kuhsiang and Huangbai are mixed in a weight ratio of 4.5:3:1.5:0.75, respectively crushed to 90 mesh, 7 times the weight of 75% ethanol solution is added, refluxed at 70℃ for 3 hours, filtered and concentrated under reduced pressure to a relative density of 1.12 extract, the Chinese herbal medicine extract is obtained Preparation Example Three: Preparation of Chinese herbal medicine extract: Honeysuckle, Huangqi, Kuhsiang and Huangbai are mixed in a weight ratio of 6:4:2:1, respectively crushed to 100 mesh, 8 times the weight of 75% ethanol solution is added, refluxed at 70℃ for 3 hours, filtered and concentrated under reduced pressure to a relative density of 1.12 extract, the Chinese herbal medicine extract is obtained. Example One

[0023] The Chinese herbal medicine composition antibacterial nanoemulsion provided by the embodiment of the application comprises the following components in parts by weight: Chinese herbal medicine extract obtained in Preparation Example One 20 parts, ethanol 15 parts, propylene glycol 10 parts, olive oil 3 parts, tea seed oil 2 parts, Tween-80 5 parts, Span-80 3 parts, n-butanol 3 parts, xanthan gum 1 part, and the balance is distilled water. The Chinese herbal medicine extract is prepared from honeysuckle, Huangqi, Kuhsiang and Huangbai, and the weight ratio of the four is 3:2:1:0.5.

[0024] The solvent is a combination of ethanol and propylene glycol, and the weight ratio is 3:1; The emulsifier is a combination of Tween-80 and Span-80, and the weight ratio is 2:1 The co-emulsifier is n-butanol, the oil phase carrier is a combination of olive oil and tea seed oil, and the co-stabilizer is xanthan gum.

[0025] Please refer to the attached Figure 1 A preparation method of a Chinese herbal medicine composition antibacterial nanoemulsion, comprising the following steps: Step One, honeysuckle, Huangqi, Kuhsiang and Huangbai are respectively crushed to 80 mesh, and are ready for use; Step two, after the raw materials are crushed, the raw materials are mixed according to the weight ratio to obtain a Chinese herbal medicine raw material mixture, then a solvent is added, the amount of the solvent added is 5 times the total weight of the raw materials, and the reflux extraction method is used at 65°C for 2.5 hours to obtain an extract; Step three, the obtained extract is filtered, and the filtrate is concentrated under reduced pressure at 40°C to a relative density of 1.10 to obtain a Chinese herbal medicine extract; Step four, after obtaining the Chinese herbal medicine extract, the Chinese herbal medicine extract, solvent, emulsifier, co-emulsifier and oil phase carrier are mixed, stirred at a speed of 3000 r / min for 10 minutes to obtain a mixed solution A; Step five, a co-stabilizer is added to the mixed solution A, and stirring is continued at a speed of 3000 r / min for 5 minutes to obtain a mixed solution B; Step six, the mixed solution B is slowly added to a water phase at 50°C, the amount of the water phase added is 3 times the total weight of the mixed solution B, and stirring is continued at a speed of 2000 r / min for 20 minutes, and then cooled to room temperature to obtain a Chinese herbal medicine composition antibacterial nanoemulsion. The nanoemulsion cooled to room temperature is stored at 1°C. Example two

[0026] A Chinese herbal medicine composition antibacterial nanoemulsion, comprising the following components by weight: 30 parts of the Chinese herbal medicine extract obtained in Preparation Example two; 32 parts of solvent is 24 parts of ethanol and 8 parts of propylene glycol; 8 parts of oil phase carrier is 4 parts of tea seed oil and 4 parts of soybean oil; 12 parts of emulsifier is 8 parts of Tween-80 and 4 parts of Span-80; 5 parts of co-emulsifier is 5 parts of ethylene glycol monomethyl ether; 2 parts of co-stabilizer is 1 part of xanthan gum and 1 part of gum arabic; The remaining amount of water phase is phosphate buffer.

[0027] Please refer to the attached Figure 1 A preparation method of a Chinese herbal medicine composition antibacterial nanoemulsion, comprising the following steps: Step one, honeysuckle, scute, kuh-seng and huang-bai are crushed to 90 mesh respectively and reserved; Step two, a Chinese herbal medicine raw material mixture is obtained by mixing, then a solvent is added, which is consistent with that in Example one, the amount of the solvent added is 6.5 times the total weight of the raw materials, and the reflux extraction method is used at 70°C for 3 hours to obtain an extract; Step three, the obtained extract is filtered, and the filtrate is concentrated under reduced pressure at 45°C to a relative density of 1.12 to obtain a Chinese herbal medicine extract; Step four, after obtaining the Chinese herbal medicine extract, mix the Chinese herbal medicine extract, solvent, emulsifier, co-emulsifier and oil phase carrier, stir at a speed of 3500 r / min for 12 minutes to obtain a mixed solution A; Step five, add co-stabilizer to the mixed solution A, continue to stir at a speed of 3500 r / min for 6 minutes to obtain a mixed solution B; Step six, slowly add the mixed solution B to the water phase at 55℃, the water phase is added in an amount of 4 times the total weight of the mixed solution B, while stirring at a speed of 2500 r / min for 25 minutes, and then cool to room temperature to obtain the Chinese herbal medicine composition antibacterial nanoemulsion. The nanoemulsion cooled to room temperature is stored at 2℃. Example three

[0028] A Chinese herbal medicine composition antibacterial nanoemulsion, comprising the following components by weight: Chinese herbal medicine extract obtained in Preparation Example three 40 parts; Solvent 40 parts, ethanol 30 parts and propylene glycol 10 parts; Oil phase carrier 12 parts, olive oil 6 parts and tea seed oil 6 parts; Emulsifier 15 parts, 10 parts of Tween-80 and 5 parts of Span-80; Co-emulsifier 8 parts, n-butanol 8 parts; Co-stabilizer 3 parts, xanthan gum 2 parts and sodium carboxymethyl cellulose 1 part; The balance of the water phase is distilled water.

[0029] Please refer to the attached Figure 1 A method for preparing a Chinese herbal medicine composition antibacterial nanoemulsion, comprising the following steps: Step one, grind honeysuckle, scute, sophora, and cork tree to 100 mesh respectively and reserve; Step two, mix to obtain a Chinese herbal medicine raw material mixture, then add solvent, and the solvent is added in an amount of 8 times the total weight of the raw materials, extract at 75℃ by reflux extraction for 3.5 hours to obtain an extract; Step three, filter the obtained extract, take the filtrate, and concentrate under reduced pressure at 50℃ to a relative density of 1.15 to obtain a Chinese herbal medicine extract; Step four, after obtaining the Chinese herbal medicine extract, mix the Chinese herbal medicine extract, solvent, emulsifier, co-emulsifier and oil phase carrier, stir at a speed of 4000 r / min for 15 minutes to obtain a mixed solution A; Step five, add co-stabilizer to the mixed solution A, continue to stir at a speed of 4000 r / min for 8 minutes to obtain a mixed solution B; Step six, slowly add the mixed solution B to the aqueous phase at 60°C, the amount of the aqueous phase added is 5 times the total weight of the mixed solution B, stir at a speed of 3000 r / min for 30 minutes, cool to room temperature, and obtain the antibacterial nanoemulsion of the Chinese herbal medicine composition. After cooling to room temperature, the nanoemulsion is stored at 4°C.

[0030] Comparative Example 1: The difference from Example 2 is that the nanoemulsion does not contain honeysuckle extract, and the other components and preparation methods are the same as those of Example 2.

[0031] Comparative Example 2: The difference from Example 2 is that the nanoemulsion does not contain the scutellaria baicalensis extract, and the other components and preparation methods are the same as those of Example 2.

[0032] Comparative Example 3: The difference from Example 2 is that the nanoemulsion does not contain Sophora flavescens extract, and the other components and preparation method are the same as those of Example 2.

[0033] Comparative Example 4: The difference from Example 2 is that the nanoemulsion does not contain the Phellodendron amurense extract, and the other components and preparation methods are the same as those of Example 2.

[0034] Comparative Example 5: The difference from Example 2 is that the weight ratio of honeysuckle, scutellaria, sophora flavescens and phellodendron is adjusted to 1:1:1:1, and the other components and preparation method are the same as those in Example 2.

[0035] Comparative Example 6: The difference from Example 2 is that in step 2, only ethanol is used as the solvent, and propylene glycol is not used. Other components and preparation methods are the same as those in Example 2.

[0036] Comparative Example 7: The difference from Example 2 is that in step 4, only Tween-80 is used as the emulsifier, and Span-80 is not used. The other components and preparation methods are the same as those in Example 2. Comparative Example 8: The difference from Example 2 is that the temperature of the aqueous phase in step 6 is 25° C., and the other components and preparation methods are the same as those in Example 2.

[0037] Common pathogens in aquaculture, Aeromonas hydrophila, Edwardsiella and Vibrio, were selected, and the antibacterial effects of the samples prepared in Examples 1 to 3 and Comparative Examples 1 to 8 of the present invention were tested using the plate count method. Test method: the pathogenic bacteria in logarithmic growth phase were diluted to 106CFU / mL, 1mL of the bacterial solution was added into the medium containing different concentrations of samples, so that the final concentration of the sample was 0.1mg / mL, 0.5mg / mL and 1mg / mL respectively, a blank control group was set up, only the bacterial solution and the medium were added, 3 replicates were set up for each group, after 24 hours of culture at 37℃, plate counting was performed, and the inhibition rate was calculated. Inhibition rate = (number of bacteria in the blank control group-number of bacteria in the experimental group) / number of bacteria in the blank control group x 100%.

[0038] Table 1: Inhibition control table

[0039] According to Table 1, the stability of examples 1 to 3 is obviously better than that of comparative examples 1 to 8, the stability of comparative examples 1 to 4 decreases due to the lack of one Chinese herbal medicine extract, the stability of comparative example 5 is also affected due to the change of the weight ratio of four Chinese herbal medicines, the stability of comparative examples 6 to 8 decreases more obviously due to the change of the corresponding solvent and emulsifier in the preparation method and the temperature of the water phase.

[0040] The antibacterial nanoemulsion of the Chinese herbal medicine composition of the application is prepared from honeysuckle, scutellaria, sophora and phellodendron as raw materials, through a formula and a preparation method, and has significant antibacterial effect, good stability and high bioavailability.

[0041] Although the embodiments of the application have been shown and described, it is to be understood that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the application, the scope of the application being defined by the appended claims and their equivalents.

Claims

1. A Chinese herbal medicine antibacterial nanoemulsion, characterized in that: It comprises the following components in parts by weight: 20-40 parts of Chinese herbal medicine extract; 25-40 parts of solvent; 5-12 parts of oil phase carrier; 8-15 parts of emulsifier; 3-8 parts of co-emulsifier; 1-3 parts of co-stabilizer; The balance was the aqueous phase.

2. The antibacterial nanoemulsion of a Chinese herbal medicine composition according to claim 1, characterized in that: The Chinese herbal medicine extract is prepared from honeysuckle, scutellaria, sophora flavescens and phellodendron amurense, and the weight ratio of the honeysuckle, scutellaria, sophora flavescens and phellodendron amurense is (3-6): (2-4): (1-2): (0.5-1).

3. The antibacterial nanoemulsion of a Chinese herbal medicine composition according to claim 1, characterized in that: The solvent is one or more combinations of ethanol, propylene glycol or glycerol; The oil phase carrier is one or more combinations of olive oil, camellia oil or soybean oil; The auxiliary stabilizer is one or more combinations of xanthan gum, gum arabic or sodium carboxymethyl cellulose.

4. The antibacterial nanoemulsion of a Chinese herbal medicine composition according to claim 1, characterized in that: The emulsifier is one or more combinations of Tween-80, Span-80 or polyoxyethylene castor oil; The auxiliary emulsifier is any one of n-butanol, ethylene glycol monomethyl ether or propylene glycol methyl ether.

5. The antibacterial nanoemulsion of a Chinese herbal medicine composition according to any one of claims 1 to 4, characterized in that: The nanoemulsion particle size is 80-200 nm, and the polydispersity index is ≤0.

25.

6. A method for preparing an antibacterial nanoemulsion of a Chinese herbal medicine composition, characterized in that: The antibacterial nanoemulsion of a Chinese herbal medicine composition according to any one of claims 1 to 5 comprises the following steps: Step 1: Grind honeysuckle, scutellaria, sophora flavescens, and phellodendron into 80-100 mesh respectively and set aside; Step 2: After the raw materials in step 1 are crushed, they are mixed to obtain a mixture of Chinese herbal raw materials, and then a solvent is added in an amount of 5-8 times the total weight of the raw materials. The mixture is extracted by reflux extraction at 65-75° C. for 2.5-3.5 hours to obtain an extract; Step 3, filtering the obtained extract, taking the filtrate, and concentrating it under reduced pressure at 40-50° C. to an extract with a relative density of 1.10-1.15 to obtain a Chinese herbal medicine extract; Step 4: After obtaining the Chinese herbal extract, the Chinese herbal extract, solvent, emulsifier, co-emulsifier and oil phase carrier are mixed, and stirred at a speed of 3000-4000 r / min for 10-15 minutes to obtain a mixed solution A; Step 5: Add a stabilizer to the mixed solution A, and continue stirring at a speed of 3000-4000 r / min for 5-8 minutes to obtain a mixed solution B; Step 6: slowly add the mixed solution B to the aqueous phase at 50-60°C, the amount of aqueous phase added is 3-5 times the total weight of the mixed solution B, stir at a speed of 2000-3000 r / min for 20-30 minutes, and cool to room temperature to obtain the Chinese herbal medicine composition antibacterial nanoemulsion.

7. The method for preparing the antibacterial nanoemulsion of a Chinese herbal medicine composition according to claim 6, characterized in that: In the step 2, the solvent is a combination of ethanol and propylene glycol, and the weight ratio of ethanol to propylene glycol is 3:

1.

8. The method for preparing the antibacterial nanoemulsion of a Chinese herbal medicine composition according to claim 6, characterized in that: In the step 4, the emulsifier is a combination of Tween-80 and Span-80, and the weight ratio of Tween-80 to Span-80 is 2:1, and the co-emulsifier is n-butanol.

9. The method for preparing the antibacterial nanoemulsion of a Chinese herbal medicine composition according to claim 6, characterized in that: The aqueous phase is any one of distilled water or phosphate buffer.

10. The method for preparing the antibacterial nanoemulsion of a Chinese herbal medicine composition according to claim 6, characterized in that: In the step 6, the nanoemulsion is stored at 1-4° C. after being cooled to room temperature.

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