CN106173806B - SiO (silicon dioxide)2Preparation method and application of artemisinin antibacterial nanoparticles - Google Patents
SiO (silicon dioxide)2Preparation method and application of artemisinin antibacterial nanoparticles Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/358—Inorganic compounds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/24—Inorganic compounds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
Abstract
The invention belongs to the field of antibacterial agents and food additives, and particularly relates to SiO2Preparation method and application of artemisinin antibacterial nanoparticles are provided. Using nano SiO2Adsorbing artemisinin to prepare antibacterial nanoparticles, and combining with plasma for assisting treatment, the method can improve solubility of artemisinin in water and has a certain slow release effect, thereby improving bioavailability of artemisinin and achieving the purposes of efficient utilization and long-acting antibacterial.
Description
Technical Field
The invention belongs to the field of antibacterial agents and food additives, and particularly relates to SiO2Preparation and application of artemisinin antibacterial nanoparticles.
Background
Artemisia annua (Artemisia annua L.) belonging to genus Artemisia of family Compositae; the artemisinin is a sesquiterpene lactone medicine with peroxy groups extracted from stems and leaves of plant artemisia annua by Chinese researchers in 1972, and is an effective component capable of treating malaria; the artemisinin has various pharmacological actions such as malaria resistance, immunoregulation, tumor resistance, antibiosis and the like; however, because artemisinin has water insolubility, the antibacterial activity of artemisinin is greatly reduced in practical application, and the research focuses on solving the problem of water solubility of artemisinin, improving the water dispersibility of artemisinin, further improving the antibacterial activity of artemisinin and increasing the bioavailability of artemisinin.
At present, there are many patent applications for artemisinin in China. Chinese patent CN101293889A discloses a water-soluble artemisinin derivative and a preparation method thereof, which have good water solubility, are easy to prepare, have no toxic or side effect on normal cells and have patent medicine prospect; chinese patent CN102727440A proposes an artemisinin ultrafine powder and a preparation method thereof, wherein the artemisinin ultrafine powder has spheroidal particles, good water dispersibility, narrow particle size distribution and high bioavailability; chinese patent CN102755316A discloses an application of artemisinin and its derivatives in preparing drugs for treating hepatitis C virus, belonging to the field of medicine; chinese patent CN103948585A discloses the application of artemisinin and its derivatives in the preparation of drugs for preventing and treating neurological diseases, which brings a new research direction for the treatment and prevention of neurological diseases; chinese patent CN104906084A discloses an application of artemisinin and its derivatives in preparing drugs for preventing and treating ophthalmic vascular diseases and a pharmaceutical composition, belonging to the application in medicine.
Nano SiO2Is white powder, has large specific surface area and better performanceHas good adsorption property, and can be widely used in the fields of food, antibacterial material and medicine. In the field of antibacterial materials, nano SiO is utilized2The nano antibacterial agent has the advantages of large specific surface area, multi-mesoporous structure on the surface, physiological inertia and super strong adsorption capacity, is combined with antibacterial active ingredients to develop a high-efficiency, durable and broad-spectrum antibacterial nano antibacterial agent, and can be widely used in the fields of food industry, medical health and the like. In the food industry, nano SiO is added2The food packaging bag can play a role in keeping fruits and vegetables fresh; nano SiO2The product can be used in wine production to achieve the effects of purifying and prolonging the preservation period; food-grade nano SiO2When added into granular or powdery food, the additive can keep the food particles in a loose or optimal free flowing state, and has the function of anti-caking. In the aspect of medicine, nano SiO is adopted2Has good drug slow release effect for the carrier.
At present, a plurality of patent applications about nano SiO2 exist in China. Chinese patent CN105169398A discloses a controlled release system based on mesoporous silica nanoparticles and a preparation method thereof; chinese patent CN104785214A proposes a method for preparing chitosan-coated magnetic mesoporous silica core-shell structured nanoparticles, and the product of the invention has excellent adsorption effect on methylene blue; chinese patent CN104262852A discloses that the application field of polyvinyl chloride pipes is greatly expanded by modifying polyvinyl chloride plastic particles with silica nanoparticles; chinese patent CN102424961A discloses a sol-gel coating with silica nanoparticles dispersed in a sol system and a preparation method thereof; chinese patent CN102652735A discloses porous silica nanoparticles carrying insoluble drugs, a preparation method and application thereof, belonging to the field of biological medicine.
Plasma refers to a highly ionized gas, which is a system of a large number of charged particles (ions, electrons) and neutral particles (atoms, molecules) that are ionized when the gas is subjected to external high energy (high temperature, strong electromagnetic field, radiation, etc.). The plasma reaction temperature is low, the speed is high and the environment is protected. The energy of most particles in the low-temperature plasma is slightly higher than the chemical bond energy in the polymer, and the low-temperature plasma has enough energy to cause various chemical bonds on the surface of the polymer to be broken or recombined after being accelerated by an electric field, so that the surface of the molecular material can be modified. For example, some edible films are treated by plasma techniques to alter the surface roughness of the film and increase the surface hydrophilicity of the film; and for example, the Low Density Polyethylene (LDPE) is treated by plasma, so that the adsorption quantity of Nisin is increased, and the antibacterial and fresh-keeping effects on food are achieved.
Based on nanometer SiO2The physiological inertia and high adsorbability of the artemisinin can solve the problems of poor antibacterial effect and short effective period in practical application caused by water insolubility of the artemisinin; by means of nano SiO2Adsorbing artemisinin to prepare antibacterial nanoparticles with a certain slow release effect; and the plasma assisted treatment is combined, so that the surface hydrophilicity of the artemisinin can be improved, and the solubility of the artemisinin in water is further improved, so that the utilization rate of the artemisinin is improved, and the purposes of efficient utilization and long-acting antibiosis are achieved.
Disclosure of Invention
The invention discloses a one meter SiO2Preparation and application of artemisinin antibacterial nanoparticles; using nano SiO2Adsorbing artemisinin to prepare antibacterial nanoparticles, and combining with plasma for assisting treatment, the method can improve solubility of artemisinin in water and has a certain slow release effect, thereby improving bioavailability of artemisinin and achieving the purposes of efficient utilization and long-acting antibacterial.
The invention takes Tetraethoxysilane (TEOS) and artemisinin crystals as raw materials, absolute ethyl alcohol and deionized water as cosolvent, ammonia water (25 percent, wt%) as catalyst to prepare nano SiO2The artemisinin antibacterial nano-ions are combined with plasma for assisting treatment, so that the solubility of the artemisinin can be greatly improved.
The specific preparation method of the invention is as follows: mixing absolute ethyl alcohol, deionized water and ammonia water, heating and stirring, then dropwise adding TEOS, immediately adding artemisinin crystals after dropwise adding, and maintaining the temperature to continue heating and stirring; after completion, the solvent is evaporated by vacuum drying; collecting, grinding, washing, drying and grinding the obtained powder;finally, the SiO is prepared by adopting plasma to assist treatment2Artemisinin antibacterial nanoparticles.
The volume ratio of the absolute ethyl alcohol to the deionized water to the ammonia water is as follows: 20: 5: 1.
the heating and stirring are performed by adopting a heating type magnetic stirrer and stirring for 20min at the stirring speed of 300rmp and 60 ℃.
The volume ratio of TEOS to ammonia water is 1: 1.
the addition of the tetraethyl orthosilicate (TEOS) was carried out by a dropwise addition method at a rate of 2.0mL/2 min.
The mass ratio of the artemisinin crystals to the deionized water is 5: 1.
The temperature maintaining and continuous heating and stirring means that a heating type magnetic stirrer is adopted, and stirring is carried out for 60min at the stirring speed of 300rmp and at the temperature of 60 ℃.
The vacuum drying means that the solvent evaporates: the mixture was placed in an oven at 60 ℃ and dried under vacuum for 48h to evaporate the solvent.
Washing the powder for 1 time by using absolute ethyl alcohol, washing for 2 times by using deionized water, and then washing for 1 time by using the absolute ethyl alcohol; the first step of absolute ethanol washing is to remove residual TEOS, and the second step of absolute ethanol washing is to remove water in the precipitate, so that the surface tension of the solution is reduced, and the aggregation of the powder in the drying process is reduced.
The plasma assisted treatment conditions are as follows: power 800W, time 2 min.
The invention has novel idea and special product structure, gives full play to the superiority of each component, and prepares the SiO with high stability and high antibacterial property by superposing the advantages of each component2Artemisinin antibacterial nanoparticles.
Drawings
FIG. 1 nanometer SiO2The artemisinin antibacterial nano-particles have antibacterial effect on escherichia coli.
FIG. 2 nanometer SiO2The artemisinin antibacterial nano-particles have antibacterial effect on staphylococcus aureus.
FIG. 3 nanometer SiO2Practical application of artemisinin antibacterial nanoparticles in beef (Daedan)Enterobacter).
FIG. 4 nanometer SiO2Practical application of artemisinin antibacterial nanoparticles in beef (Staphylococcus aureus).
Detailed Description
The following examples illustrate specific embodiments of the present invention, but the scope of the present invention is not limited thereto.
Example 1 SiO2Solubility of artemisinin antibacterial nanoparticles
1 materials of the experiment
2 method of experiment
① adding 40.0mL of absolute ethanol, 10.0mL of deionized water and 2.0mL of ammonia water into a reaction flask, placing on a heating magnetic stirrer, heating and stirring for 20min (60 ℃, 300rmp), then adding 2.0mL of TEOS dropwise (ensuring that all the TEOS is added within 2 min), adding 50mg of artemisinin crystal immediately after the addition, maintaining the temperature at 60 ℃, and continuing stirring for 60min at 300 rmp.
② placing the reaction bottle in an oven at 60 deg.C for 48h, vacuum drying to evaporate the solvent, collecting the powder, grinding, washing, further drying, grinding, and collecting.
③ and finally processing with plasma under the conditions of 800W power and 2min time to obtain SiO2Artemisinin antibacterial nanoparticles.
④ it is specifically stated that the mark without artemisinin crystals added during the manufacturing process is blank SiO2Nanoparticles.
3 SiO2Determination of solubility of artemisinin antibacterial nanoparticles
Weigh the SiO once prepared (with 50mg of artemisinin crystals added) and dried to constant weight2Artemisinin antibacterial nanoparticles, marked as W, were added to 40.0mL distilled water in a 50mL beaker, the sample was dissolved with shaking and transferred to a 50mL centrifuge tube, centrifuged with a lid (5000r/min, 10min), and the supernatant was decanted. Adding 40mL of distilled water to fully dissolve the sample, and transferring into a centrifugeThe tube was covered, centrifuged (5000r/min, 10min) and the supernatant decanted, and the precipitate washed with a small amount of distilled water into an evaporation dish of known mass and dried in an oven at 100 ℃ to constant weight. The control group was: 50mg artemisinin crystal and (W-50) mg blank SiO2The nano particles are operated in the same way as above. The solubility calculation formula is as follows:
wherein W is the mass of the sample; w1The mass of the evaporating dish; w2The total mass of the evaporating dish and insoluble matter dried to constant weight.
4 SiO2Solubility of artemisinin antibacterial nanoparticles
Through calculation and analysis, SiO2The solubility of artemisinin in the artemisinin antibacterial nanoparticles is greatly improved, and is about 30% higher than that of simple artemisinin; and plasma treated SiO2The solubility of artemisinin in the artemisinin nano particles is further improved to about 35 percent, so that the nano SiO2The introduction and the assistant treatment of the plasma greatly improve the water solubility and the bioavailability of the artemisinin.
Example 2 SiO2Determination of antibacterial performance of artemisinin antibacterial nanoparticles
1 materials of the experiment
SiO2Artemisinin antibacterial nano particle-plasma, SiO2Artemisinin antibacterial nano particles and artemisinin crystal strains: escherichia coli (Escherichia coli), Staphylococcus aureus (Staphylococcus aureus)
2 method of experiment
Measuring SiO by plate colony counting method with Escherichia coli (Escherichia coli) and Staphylococcus aureus (Staphylococcus aureus) as model strains2The artemisinin antibacterial nano-particles have antibacterial performance.
Inoculating Escherichia coli and Staphylococcus aureus into liquid culture medium, respectively placing in air shaking table, shake culturing at 37 deg.C and 150rpm for 24-48 h,obtaining bacterial suspension in logarithmic growth phase. Taking a proper amount of escherichia coli and staphylococcus aureus which are saturated in the logarithmic growth phase, respectively adding the escherichia coli and the staphylococcus aureus into a test tube containing a certain amount of sterile phosphate buffer solution for dilution, so that the bacterial concentration is about 105～106CFU/mL。
Then add 4% SiO into the tube2Artemisinin antibacterial nano particle (SiO)2Artemisinin antibacterial nano-particle-plasma), adding a group of 4% artemisinin crystals as blank control, placing each test tube in an air shaking table, and carrying out oscillation reaction for 24-48 h at 37 ℃ and 150 rpm. Taking a proper amount of culture solution at two time points of 24h and 48h respectively, carrying out tenfold gradient dilution to reach a proper concentration, then transferring 100 mu L of diluted solution to a sterile solid plate culture medium, uniformly coating, and then putting into a constant temperature and humidity incubator at 37 ℃ for inverted culture. Counting plate colonies after 24-48 h, thereby counting SiO2Evaluation of antibacterial activity of artemisinin antibacterial nanoparticles. The experiment was repeated three times and the results averaged.
3 SiO2Evaluation of antibacterial performance of artemisinin antibacterial nanoparticles
Because the artemisinin crystal has water insolubility, the antibacterial performance of the simple artemisinin crystal cannot be fully exerted. As is clear from FIGS. 1 and 2, the nano SiO2And the introduction of the plasma can greatly improve the water dispersibility of the artemisinin, thereby having better antibacterial activity on escherichia coli and staphylococcus aureus. Moreover, the nano SiO can be seen from the time2Has obvious slow release effect on the artemisinin, thereby ensuring that the artemisinin can play long-acting antibacterial activity and better bactericidal effect.
Example 3 SiO2Practical application of artemisinin antibacterial nanoparticles in beef
1 materials of the experiment
SiO2Artemisinin antibacterial nano particle-plasma, SiO2Artemisinin antibacterial nano particles and artemisinin crystal strains: escherichia coli (Escherichia coli), Staphylococcus aureus (Staphylococcus aureus) beef (cut into 25X 20X 5mm size)
2 method of experiment
Measuring SiO by plate colony counting method with Escherichia coli (Escherichia coli) and Staphylococcus aureus (Staphylococcus aureus) as model strains2The artemisinin antibacterial nano-particles have antibacterial performance.
Inoculating escherichia coli and staphylococcus aureus to a liquid culture medium, and respectively placing the liquid culture medium and the staphylococcus aureus in an air shaking table to shake and culture for 24-48 h under the conditions of 37 ℃ and 150rpm to obtain bacterial suspension in a logarithmic phase. Taking a proper amount of escherichia coli and staphylococcus aureus which are saturated in the logarithmic growth phase, respectively adding the escherichia coli and the staphylococcus aureus into a test tube containing a certain amount of sterile phosphate buffer solution for dilution, so that the bacterial concentration is about 105～106CFU/mL。
Taking Escherichia coli as an example, cutting beef into 25 × 20 × 5mm, ultraviolet sterilizing (or radiation sterilizing), and treating the front and back sides for 1 hr. Soaking the treated beef sample in a solution diluted to 105～106CFU/mL of Escherichia coli suspension for 30 s.
Taking out beef sample from the bacterial liquid, air drying in sterile environment, and adding SiO 4%2A group containing 4% artemisinin crystals was used as a blank in a container of PBS liquid containing artemisinin antibacterial nanoparticles. Culturing the seeds in constant-temperature incubators at 37 ℃ for 24-72 hours respectively. Taking out beef samples at three time points of 24h, 48h and 72h respectively, then airing in a sterile environment, transferring into a sterile homogenizing bag, adding 9 times of sterile Phosphate Buffer Solution (PBS) into the sterile homogenizing bag containing the beef samples, and homogenizing. The supernatant was aspirated to determine the total number of bacteria in the beef sample. The determination method is carried out according to GB 4789.2-2010 food hygiene microbiology detection colony total number. Thereby can be applied to SiO2The antibacterial activity of the artemisinin antibacterial nanoparticles on escherichia coli in beef is evaluated. The experiment was repeated three times and the results averaged.
Evaluation of practical application of 3 SiO 2/artemisinin antibacterial nanoparticles in beef
As can be analyzed from fig. 3 and fig. 4, the SiO 2/artemisinin antibacterial nanoparticles have better effect on Escherichia coli and Staphylococcus aureus in beefAnd (4) antibacterial activity. Moreover, the nano SiO can be seen from the time2Has obvious slow release effect on the artemisinin, thereby ensuring the artemisinin to play long-acting antibacterial activity and better bactericidal effect.
Claims (7)
1. SiO (silicon dioxide)2The preparation method of the artemisinin antibacterial nano particles is characterized by comprising the following steps: using nano SiO2Adsorbing artemisinin to prepare antibacterial nanoparticles, combining with plasma for assisting treatment, improving the solubility of artemisinin in water, and having a slow release effect, thereby improving the bioavailability of artemisinin and achieving the purposes of efficient utilization and long-acting antibacterial, the specific steps are as follows: mixing absolute ethyl alcohol, deionized water and ammonia water, heating and stirring, then dropwise adding TEOS, immediately adding artemisinin crystals after dropwise adding, and maintaining the temperature to continue heating and stirring; after completion, the solvent is evaporated by vacuum drying; collecting, grinding, washing, drying and grinding the obtained powder; finally, the SiO is prepared by adopting plasma to assist treatment2Artemisinin antibacterial nanoparticles; the volume ratio of TEOS to ammonia water is 1: 1; the Tetraethoxysilane (TEOS) is added by a dropwise adding method, and the dropwise adding speed is 2.0mL/2 min; the temperature maintaining and continuous heating and stirring means that a heating type magnetic stirrer is adopted, and stirring is carried out for 60min at the stirring speed of 300rmp and at the temperature of 60 ℃.
2. An SiO as claimed in claim 12The preparation method of the artemisinin antibacterial nano particles is characterized by comprising the following steps: the volume ratio of the absolute ethyl alcohol to the deionized water to the ammonia water is as follows: 20: 5: 1; the heating and stirring are performed by adopting a heating type magnetic stirrer and stirring for 20min at the stirring speed of 300rmp and 60 ℃.
3. An SiO as claimed in claim 12The preparation method of the artemisinin antibacterial nano particles is characterized by comprising the following steps: the mass ratio of the artemisinin crystals to the deionized water is 5: 1.
4. An SiO as claimed in claim 12Artemisinin antibacterial nanoparticlesThe preparation method is characterized in that the vacuum drying enables the solvent evaporation to mean that: placing in an oven at 60 deg.C, vacuum drying for 48 hr to evaporate solvent; washing the powder for 1 time by using absolute ethyl alcohol, washing for 2 times by using deionized water, and then washing for 1 time by using the absolute ethyl alcohol; the first step of absolute ethanol washing is to remove residual TEOS, and the second step of absolute ethanol washing is to remove water in the precipitate, so that the surface tension of the solution is reduced, and the aggregation of the powder in the drying process is reduced.
5. An SiO as claimed in claim 12The preparation method of the artemisinin antibacterial nanoparticles is characterized in that the plasma assisted treatment conditions are as follows: power 800W, time 2 min.
6. An SiO as claimed in claim 12The preparation method of the artemisinin antibacterial nano particles is characterized by comprising the following steps: the concentration of ammonia was 25 wt%.
7. SiO prepared by the process as claimed in any of claims 1 to 62Application of artemisinin antibacterial nanoparticles in the field of food antibacterial and antiseptic is provided.
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