CN114532332A - Method for constructing zinc oxide/attapulgite nano composite antibacterial material by utilizing ginkgo leaf extract - Google Patents
Method for constructing zinc oxide/attapulgite nano composite antibacterial material by utilizing ginkgo leaf extract Download PDFInfo
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- A01N25/08—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing solids as carriers or diluents
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/22—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing ingredients stabilising the active ingredients
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
- A01N59/16—Heavy metals; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/30—Zinc; Compounds thereof
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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
- A61K47/02—Inorganic compounds
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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
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
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- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01G—COMPOUNDS CONTAINING METALS NOT COVERED BY SUBCLASSES C01D OR C01F
- C01G9/00—Compounds of zinc
- C01G9/02—Oxides; Hydroxides
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- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2002/00—Crystal-structural characteristics
- C01P2002/70—Crystal-structural characteristics defined by measured X-ray, neutron or electron diffraction data
- C01P2002/72—Crystal-structural characteristics defined by measured X-ray, neutron or electron diffraction data by d-values or two theta-values, e.g. as X-ray diagram
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- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2002/00—Crystal-structural characteristics
- C01P2002/80—Crystal-structural characteristics defined by measured data other than those specified in group C01P2002/70
- C01P2002/82—Crystal-structural characteristics defined by measured data other than those specified in group C01P2002/70 by IR- or Raman-data
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- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2002/00—Crystal-structural characteristics
- C01P2002/80—Crystal-structural characteristics defined by measured data other than those specified in group C01P2002/70
- C01P2002/84—Crystal-structural characteristics defined by measured data other than those specified in group C01P2002/70 by UV- or VIS- data
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a method for constructing a zinc oxide/attapulgite nano composite antibacterial material by utilizing a ginkgo leaf extract, which comprises the steps of adding attapulgite into a ginkgo leaf aqueous extract, uniformly stirring, dropwise adding a zinc salt solution, and adjusting the pH value to 7-11 by adopting sodium hydroxide to obtain a precursor mixed solution; and (3) reacting the precursor mixed solution for 30-180 min under the conditions of stirring and oil bath at the constant temperature of 55-95 ℃, and centrifugally washing and drying the obtained product to obtain the zinc oxide/attapulgite composite antibacterial material. The invention takes the ginkgo leaf extract as a reducing agent and a stabilizing agent, constructs the attapulgite-zinc oxide composite material by a one-step method, and shows excellent antibacterial performance to gram-positive bacteria and gram-negative bacteria. The preparation method has the advantages of easily-accessible raw materials, simple process, low cost, environmental friendliness, and easy mass production.
Description
Technical Field
The invention belongs to the technical field of nano composite material preparation, and particularly relates to a method for constructing a zinc oxide/attapulgite nano composite antibacterial material by utilizing a ginkgo leaf extract.
Background
Currently, common antibacterial agents can be classified into inorganic antibacterial agents, organic antibacterial agents, and natural antibacterial agents. Compared with other two antibacterial agents, the inorganic antibacterial agent has the advantages of broad spectrum, high efficiency, heat resistance, safety, environmental friendliness and the like, and becomes a research hotspot of the current novel antibacterial agent. Among them, zinc oxide is attracting much attention because of its advantages such as low cost of raw materials, long-lasting antibacterial action, safety and stability (research in stomatology 2015,31, 195). Compared with common zinc oxide, the nano zinc oxide has stronger antibacterial activity due to smaller particle size and larger specific surface area, and has higher application value in the fields of packaging materials, food additives, textiles, building materials, animal health breeding and the like (packaging engineering, 2019,40, 172). Researches prove that zinc oxide can replace antibiotics to prevent and treat livestock and poultry diarrhea (China journal of livestock husbandry 2021,57,7, Proc. 2012,24, 285). However, the zinc oxide with the nano structure has large specific surface area, high surface energy and easy agglomeration, and seriously restricts the exertion of the bioactivity and the wide application of the zinc oxide.
In recent years, composite antibacterial materials based on nano zinc oxide have attracted much attention due to their excellent antibacterial activity and broad application potential (functional materials 2018,49, 9061). The nano zinc oxide is loaded on an organic or inorganic carrier, so that the agglomeration of particles can be effectively avoided, the zinc oxide nano particles with smaller sizes can be obtained, the stability of the zinc oxide nano particles in the environment is improved, and the antibacterial activity of the nano zinc oxide is further enhanced (novel chemical materials, 2019,47 and 268). Attapulgite is a water-containing magnesium-aluminum-rich silicate clay mineral, has the characteristics of large specific surface area, rich surface functional groups and the like, has inherent advantages in the aspect of building functional materials (China science: chemistry 2018,48, 1432), and is widely applied to the fields of agriculture, chemical industry, environmental protection, adsorption separation, composite materials and the like at present. The nano zinc oxide composite material is prepared by taking attapulgite rod crystals as a carrier, so that on one hand, the growth process of nano particles can be regulated and controlled, and the high-activity zinc oxide nano particles with uniform dispersion and smaller size are obtained; on the other hand, the zinc oxide is fixedly carried in the attapulgite rod crystals, so that the zinc oxide can be prevented from being aggregated and losing efficacy in use, namely the environmental stability of the zinc oxide is enhanced. In addition, the attapulgite has the advantages of natural and easily-obtained carbon carrier materials such as graphene, low cost, easy operation and the like.
The nano zinc oxide and the composite material thereof are generally prepared by a chemical method, such as a precipitation method, a hydrothermal synthesis method, a sol-gel method and the like. The chemical preparation methods have complex process, higher cost and environmental pollution. The green synthesis method based on natural plants can reduce the use of toxic and harmful chemical reagents and the generation of side samples, and the obtained material has better biocompatibility and stability and better meets the application requirements. The ginkgo leaves contain abundant flavonoid glycoside substances and have strong effect of scavenging free radicals. It has been shown that ginkgo biloba extract can be synthesized into gold, silver, zinc oxide and other nanoparticles (optik.2017, 144,511, J Mater Sci: Mater electron. 2021,32,17154, chem. phy. let. 2018,711,100). However, the synthesis of the attapulgite-loaded zinc oxide antibacterial composite material is not reported.
Disclosure of Invention
The invention aims to provide a method for constructing a zinc oxide/attapulgite nano composite antibacterial material by utilizing a ginkgo leaf extract, which adopts the ginkgo leaf extract as a medium, can effectively avoid the generation of toxic byproducts, is green and environment-friendly, and has high activity and stability of the obtained nano composite material.
In order to achieve the above object, the present invention provides the following technical solutions:
a method for constructing a zinc oxide/attapulgite nano composite antibacterial material by utilizing a ginkgo leaf extract comprises the following steps:
1) adding the ginkgo leaf powder into distilled water under stirring, stirring in a water bath at 55-65 ℃ for 30-60 min, and filtering to obtain a ginkgo leaf water extract; the mass concentration of the ginkgo leaves in the distilled water is 5-20%.
2) Adding attapulgite powder into the ginkgo leaf aqueous extract, uniformly stirring, adding a zinc salt solution, and adjusting the pH value to 7-11 by using a sodium hydroxide solution to obtain a precursor mixed solution; the zinc salt is at least one of zinc nitrate hexahydrate, zinc sulfate heptahydrate and zinc acetate dihydrate, and the concentration of the zinc salt solution is 0.05-1 mol/L. The mass ratio of the attapulgite powder to the zinc salt is 10: 1-1: 1. The volume ratio of the ginkgo leaf aqueous extract to the zinc salt solution is 2: 1-4: 1.
3) And (3) reacting the precursor mixed solution for 30-180 min under the conditions of stirring and oil bath at the constant temperature of 55-95 ℃, and centrifugally washing and drying the obtained product to obtain the zinc oxide/attapulgite nano composite antibacterial material. The stirring speed of the precursor mixed liquid is 400-600 rpm.
The synthesis mechanism of the invention is as follows: flavonoid glycoside, terpene lactone, organic acid, alkylphenol, protein, polysaccharide and other phytochemicals in the ginkgo biloba extract are key factors for reducing and stabilizing the zinc oxide nanoparticles.
FIG. 1 is an infrared spectrum of the zinc oxide/attapulgite nano composite material prepared by the invention, which is 3419cm-1And 1452cm-1The shift in absorption peaks can be attributed to N-H stretching vibration of the amide group and C-N stretching vibration of the aromatic amine group; 1647 and 640cm-1Is shifted from the absorption peak and 1032cm-1The peak is enhanced by extractingThe stretching vibration of C-OH and C-H of protein, polyphenol and olefin groups in the extract is caused, which shows that flavonoid glycoside, alkylphenol, organic acid, protein and the like in the ginkgo extract participate in the reduction and stabilization of the nanoparticles. The polar hydroxyl groups on the molecules of these compounds contain electron-rich oxygen atoms and are capable of interacting with positively charged zinc ions, while the carboxyl and amino groups in the active molecules are key factors in stabilizing the nanoparticles. Furthermore, 469cm-1The peak enhancement indicates the formation of zinc oxide. Fig. 2 and 3 are ultraviolet absorption and XRD spectra of the nanocomposite, respectively, in which absorption peaks and diffraction peaks of zinc oxide appear, indicating that zinc oxide nanoparticles were successfully prepared.
The zinc oxide/attapulgite nano composite material shows excellent antibacterial performance to gram-positive bacteria and gram-negative bacteria.
In conclusion, the zinc oxide/attapulgite nano composite antibacterial material prepared by mediating and synthesizing the ginkgo leaf water extract has the following advantages: attapulgite is introduced as a carrier to obtain zinc oxide nano-particles with smaller particle size, so that the antibacterial activity of the composite material is improved; the ginkgo leaf extract is used as a reducing agent and a stabilizing agent, does not relate to the generation of toxic chemical substances and byproducts, does not cause environmental pollution, and the obtained nano-particles have high safety, good stability and excellent broad-spectrum antibacterial activity. The preparation method has the advantages of convenient material acquisition, simple process operation, low cost, environmental protection and easy mass production.
Drawings
FIG. 1 is a FTIR chart of a zinc oxide/attapulgite nanocomposite prepared in example 2 of the present invention;
FIG. 2 is a UV-vis diagram of the zinc oxide/attapulgite nanocomposite prepared in example 2 of the present invention;
FIG. 3 is an XRD pattern of the zinc oxide/attapulgite nanocomposite prepared in example 2 of the present invention.
Detailed Description
The method for constructing the zinc oxide/attapulgite nano composite antibacterial material by using the ginkgo biloba extract according to the invention is explained in detail by combining with the specific embodiment.
Example 1
1) Adding 5g folium Ginkgo powder into 100mL distilled water under stirring, stirring in 55 deg.C water bath for 30min, and filtering to obtain folium Ginkgo water extract; 2) weighing 0.81g of attapulgite powder, adding 40mL of ginkgo leaf aqueous extract, uniformly stirring, dropwise adding 20mL of 0.05 mol/L-concentration zinc nitrate hexahydrate solution, and adjusting the pH value to 7 by adopting 1mol/L sodium hydroxide to obtain precursor mixed solution; 3) and (3) reacting the precursor mixed solution for 60min under the conditions of mechanical stirring at 400rpm and constant-temperature oil bath at 80 ℃, centrifuging the obtained product, washing for three times, and drying to obtain the zinc oxide/attapulgite nano composite antibacterial material.
Example 2
1) Adding 10g folium Ginkgo powder into 100mL distilled water under stirring, stirring in 60 deg.C water bath for 30min, and filtering to obtain folium Ginkgo water extract; 2) weighing 0.54g of attapulgite powder, adding 50mL of ginkgo leaf aqueous extract, uniformly stirring, dropwise adding 20mL of 0.1mol/L heptahydrate zinc sulfate solution, and adjusting the pH value to 9 by adopting 1mol/L sodium hydroxide to obtain precursor mixed solution; 3) and (3) reacting the precursor mixed solution for 120min under the conditions of mechanical stirring at 600rpm and constant-temperature oil bath at 60 ℃, centrifuging the obtained product, washing for three times, and drying to obtain the zinc oxide/attapulgite nano composite antibacterial material.
Example 3
1) Adding 10g folium Ginkgo powder into 100mL distilled water under stirring, stirring in 60 deg.C water bath for 40min, and filtering to obtain folium Ginkgo water extract; 2) weighing 0.81g of attapulgite powder, adding 60mL of ginkgo leaf aqueous extract, uniformly stirring, dropwise adding 20mL of 0.25mol/L zinc nitrate hexahydrate solution, and adjusting the pH value to 8 by adopting 1mol/L sodium hydroxide to obtain precursor mixed solution; 3) and (3) reacting the precursor mixed solution for 30min under the conditions of mechanical stirring at 500rpm and constant-temperature oil bath at 95 ℃, centrifuging the obtained product, washing for three times, and drying to obtain the zinc oxide/attapulgite nano composite antibacterial material.
Example 4
1) Adding 15g folium Ginkgo powder into 100mL distilled water under stirring, stirring in 65 deg.C water bath for 50min, and filtering to obtain folium Ginkgo water extract; 2) weighing 1.2g of attapulgite powder, adding 70mL of ginkgo leaf aqueous extract, uniformly stirring, dropwise adding 20mL of 0.5mol/L zinc acetate dihydrate solution, and adjusting the pH value to 10 by adopting 1mol/L sodium hydroxide to obtain precursor mixed solution; 3) the precursor mixed solution reacts for 90min under the conditions of mechanical stirring at 600rpm and constant-temperature oil bath at 70 ℃, and the obtained product is centrifuged, washed for three times and dried
Drying to obtain the zinc oxide/attapulgite nano composite antibacterial material.
Example 5
1) Adding 20g folium Ginkgo powder into 100mL distilled water under stirring, stirring in 60 deg.C water bath for 60min, and filtering to obtain folium Ginkgo water extract; 2) weighing 1.62g of attapulgite powder, adding 80mL of ginkgo leaf aqueous extract, uniformly stirring, dropwise adding 20mL of 1mol/L zinc nitrate hexahydrate solution, and adjusting the pH value to 11 by adopting 1mol/L sodium hydroxide to obtain precursor mixed solution; 3) and (3) reacting the precursor mixed solution for 180min under the conditions of mechanical stirring at 600rpm and constant-temperature oil bath at 55 ℃, centrifuging the obtained product, washing for three times, and drying to obtain the zinc oxide/attapulgite nano composite antibacterial material.
The antibacterial performance of the zinc oxide/attapulgite nanocomposite prepared in examples 1-5 was evaluated by measuring the Minimum Inhibitory Concentration (MIC) using the agar dilution method, and the test procedure was as follows: dispersing the sterilized samples to be tested into agar culture media at 40-50 ℃ according to different concentrations, and cooling to obtain sample plates; setting the concentration of 1-2 muL to 104CFU/mL of fresh bacterial suspension was inoculated onto the sample plate at 3 different positions, while positive and negative control plates were prepared, and the inoculated plates were then incubated in a 37 ℃ incubator for 18-24 h. Bacteria were considered to be completely inhibited if no colony grew at all in the sample plates after completion of the incubation, wherein the minimum sample concentration to completely inhibit bacterial growth was the MIC. The MICs of the zinc oxide/attapulgite nanocomposites of examples 1-5 for E.coli and Staphylococcus aureus are shown in Table 1.
TABLE 1 MIC (mg/mL) of the Zinc oxide/Attapulgite nanocomposites in examples 1-5 for E.coli and Staphylococcus aureus
Claims (6)
1. A method for constructing a zinc oxide/attapulgite nano composite antibacterial material by utilizing a ginkgo leaf extract is characterized by comprising the following steps of:
1) adding the ginkgo leaf powder into distilled water under stirring, stirring in a water bath at 55-65 ℃ for 30-60 min, and filtering to obtain a ginkgo leaf water extract;
2) adding attapulgite powder into the ginkgo leaf aqueous extract, uniformly stirring, adding a zinc salt solution, and adjusting the pH value to 7-11 by using a sodium hydroxide solution to obtain a precursor mixed solution;
3) and (3) reacting the precursor mixed solution for 30-180 min under the conditions of stirring and oil bath at the constant temperature of 55-95 ℃, and centrifugally washing and drying the obtained product to obtain the zinc oxide/attapulgite nano composite antibacterial material.
2. The method for constructing the zinc oxide/attapulgite nano-composite antibacterial material by utilizing the ginkgo biloba extract as claimed in claim 1, is characterized in that: in the step 1), the mass concentration of the ginkgo leaves in the distilled water is 5-20%.
3. The method for constructing the zinc oxide/attapulgite nano-composite antibacterial material by utilizing the ginkgo biloba extract as claimed in claim 1, is characterized in that: in the step 2), the zinc salt is at least one of zinc nitrate hexahydrate, zinc sulfate heptahydrate and zinc acetate dihydrate, and the concentration of the zinc salt solution is 0.05-1 mol/L.
4. The method for constructing the zinc oxide/attapulgite nano-composite antibacterial material by utilizing the ginkgo biloba extract as claimed in claim 1, is characterized in that: in the step 2), the mass ratio of the attapulgite powder to the zinc salt is 10: 1-1: 1.
5. The method for constructing the zinc oxide/attapulgite nano-composite antibacterial material by utilizing the ginkgo biloba extract as claimed in claim 1, is characterized in that: in the step 2), the volume ratio of the ginkgo leaf aqueous extract to the zinc salt solution is 2: 1-4: 1.
6. The method for constructing the zinc oxide/attapulgite nano-composite antibacterial material by utilizing the ginkgo biloba extract as claimed in claim 1, is characterized in that: in the step 3), the stirring speed of the precursor mixed liquid is 400-600 rpm.
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