CN113996800B - Method for preparing nano-silver based on gynura bicolor extract - Google Patents
Method for preparing nano-silver based on gynura bicolor extract Download PDFInfo
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- CN113996800B CN113996800B CN202111259506.0A CN202111259506A CN113996800B CN 113996800 B CN113996800 B CN 113996800B CN 202111259506 A CN202111259506 A CN 202111259506A CN 113996800 B CN113996800 B CN 113996800B
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Abstract
The invention discloses a method for preparing nano-silver based on a gynura bicolor extract. Mixing the ethanol extract of the gynura bicolor with the silver ion solution, and reacting to generate the nano silver. The invention utilizes the gynura bicolor extract to reduce silver ions and prepare the simple substance nano silver. The method can prepare the nano silver particles with the particle size of 3-33 nm. The method for preparing the nano silver particles by utilizing the gynura bicolor extracting solution is easy to implement, strong in operability and low in cost, the gynura bicolor extracting solution is simultaneously used as a reducing agent and a stabilizing agent, other chemical components are not required to be added in the reaction process, a large number of nano silver particles are prepared in a short time, the method is an environment-friendly nano silver preparation method, and the prepared nano silver solution has high antibacterial effect.
Description
The technical field is as follows:
the invention belongs to the fields of biotechnology and antibacterial application, and particularly relates to a method for preparing nano-silver based on an gynura bicolor extract.
Background art:
the Silver nanoparticles (Ag-NPs for short) are called as a new generation antibacterial agent in twenty-first century, and have the advantages of broad-spectrum and efficient antibacterial performance, lasting antibacterial property, strong permeability, safety, no toxicity, restoration and regeneration, and difficulty in causing the drug resistance of microorganisms. Nano silver is widely used in various medical materials, such as wound dressings, masks for surgeons, medical catheters, medical surgical instruments, implants for human tissues and the like. Based on the excellent antibacterial property of the nano-silver, the nano-silver antibacterial material has wide application prospect and has wide market demands in the fields of articles for daily use, medical treatment and health, building materials, electronic industry and the like.
Common methods for preparing nano silver include chemical reduction, physical reduction, biological reduction and the like. The chemical method comprises the following steps: chemical methods such as a liquid-phase chemical reduction method, an adsorption in-situ reduction method, an ethylene glycol hydrothermal reduction method, an electrochemical method, a photochemical reduction method and the like; the physical method comprises the following steps: a reduction ball milling method, an ultrasonic wave reduction method, a microwave-assisted reduction method, a photon reduction method, an evaporation condensation method and an atomization method; the biological reduction method adopts natural plant extract as a reducing agent and a stabilizing agent to prepare the nano-silver. The chemical method is common, but the reducing agent and the stabilizing agent are toxic reagents, which seriously pollute the environment, and the reaction conditions need to be strictly controlled, so the preparation technology has very high requirements.
Gynura bicolor (Gynura bicolor D.C) is called Gynura bicolor, notoginseng, herba violae and the like, belongs to perennial root herbaceous plants in sanchi of Compositae, and is mostly distributed in southern areas of China. The pharmaceutical experiment shows that the semiaquilegia purpurea has the functions of clearing away heat and toxic materials, promoting blood circulation, stopping bleeding, resisting inflammation, oxidation, tumors, aging, regulating immunity, improving hematopoiesis, reducing blood sugar and blood fat and the like.
The invention content is as follows:
the invention aims to provide a method for preparing nano-silver based on an gynura bicolor extracting solution, which is simple, convenient and easy to implement, strong in operability, low in cost and environment-friendly, and explores the antibacterial effect of the nano-silver.
The method for preparing nano-silver based on the gynura bicolor extract comprises the following steps:
mixing the ethanol extract of the gynura bicolor with the silver ion solution, and reacting to generate the nano silver.
Preferably, the gynura bicolor ethanol extract is obtained by drying and crushing the leaf part of gynura bicolor, reflux-extracting gynura bicolor powder with an ethanol water solution, performing ultrasonic treatment, and filtering.
Preferably, the method comprises the following specific steps:
(1) Washing leaf part of gynura bicolor with water, drying, and pulverizing with a pulverizer to obtain gynura bicolor powder;
(2) According to mass-to-volume ratio =1g: adding the gynura bicolor powder into an ethanol aqueous solution with the volume fraction of 75% in a proportion of 20-30 ml, carrying out hot reflux for 2h at 80 ℃, continuously stirring, cooling to room temperature, carrying out ultrasonic treatment under an ice bath condition, and filtering to obtain a gynura bicolor ethanol extracting solution;
(3) 2 to 10mmol/L of AgNO 3 The solution and the gynura bicolor ethanol extract are mixed according to the volume ratio of 2-10: 1, heating and stirring at 75-95 ℃ for reaction for 15-40 min to prepare the nano silver solution.
Preferably, the washing of the gynura bicolor leaf part with water is that the gynura bicolor leaf part is washed with ultrapure water for 3 times.
Preferably, the drying is drying at 65 ℃ for 1-2 d.
Preferably, the gynura bicolor powder is pulverized by a pulverizer and then passes through a 40-mesh sieve.
Preferably, the ultrasonic treatment under the ice bath condition is carried out under the ice bath condition, the ultrasonic power is 300W, and ultrasonic treatment is carried out for 30min for 2sec and 4 sec.
Preferably, the ethanol extract of gynura bicolor obtained by filtration is filtered by a 0.22 mu M filter membrane.
Preferably, the step (3) is to use AgNO with the concentration of 10mmol/L 3 The volume ratio of the solution to the gynura bicolor extract is 5:1, heating and stirring at 95 ℃ in a dark place for 20min, and preparing the nano-silver solution.
The invention utilizes the gynura bicolor extract to reduce silver ions and prepare the simple substance nano silver. The method can prepare the nano silver particles with the particle size of 3-33 nm. The invention utilizes the gynura bicolor extract to prepare the nano silver particles, the preparation method is easy to implement, the operability is strong, the cost is low, the gynura bicolor extract is simultaneously used as a reducing agent and a stabilizing agent, other chemical components are not required to be added in the reaction process, a large number of nano silver particles are prepared in a short time, the preparation method is an environment-friendly nano silver preparation method, and the synthesized nano silver solution has high-efficiency antibacterial performance.
The method for synthesizing the nano-silver by utilizing the gynura bicolor extracting solution has high safety and strong operability. The method for synthesizing the nano-silver by reducing the plant extract has the advantages of local material availability, mild reaction conditions, low cost, low toxicity, difficult agglomeration of products, less pollution and the like. At present, the research on the reduction synthesis of nano-silver by utilizing the gynura bicolor extracting solution is little in China, the invention is a method for preparing a large number of nano-silver particles in a short time by taking the gynura bicolor extracting solution as a reducing agent and a stabilizing agent at the same time without adding other chemical components, and the synthesized nano-silver solution has high-efficiency antibacterial performance.
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FIG. 1 is a diagram showing the UV/Vis absorption of the nano-silver solution (b), the silver nitrate solution and the gynura bicolor extract in example 1;
FIG. 2 is a transmission electron micrograph (A) and a particle size distribution (B) of the nanosilver solution of example 1;
FIG. 3 is a graph showing the bacteriostatic effect of the nano-silver solution synthesized in example 1 on Escherichia coli;
FIG. 4 is a graph showing the bacteriostatic effect of the nano-silver solution synthesized in example 1 on Staphylococcus aureus;
FIG. 5 is a diagram of the bacteriostatic effect of the nano-silver solution synthesized in example 1 on Pseudomonas aeruginosa.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Example 1:
picking leaf parts of gynura bicolor, cleaning with ultrapure water for 3 times, drying at 65 ℃ for 2 days, crushing by a crusher, and sieving by a 40-mesh sieve to obtain gynura bicolor powder. Weighing 3g of gynura bicolor powder, adding 90ml of 75% ethanol aqueous solution with volume fraction, performing hot reflux at 80 ℃ for 2h, continuously stirring, and cooling to room temperature; carrying out ice bath ultrasonic 30min under the conditions of 300W, ultrasonic 2sec and stopping 4sec, roughly filtering the extracting solution by using 200-mesh gauze, and filtering the extracting solution by using a 0.22 mu M filter membrane to obtain the gynura bicolor extracting solution. AgNO with the concentration of 10mmol/L 3 The volume ratio of the solution to the gynura bicolor extract is 5:1, and keeping away from light at 95 DEG CHeating and stirring to react for 20min to prepare the nano silver solution.
FIG. 1 shows the UV-VIS absorption curves of the synthesized nano-silver solution, silver nitrate extract and gynura bicolor extract measured at 350-650 nm. Wherein: a is a gynura bicolor extract solution; b is nano silver solution synthesized by reaction; c is silver nitrate solution. The figure shows that the synthesized nano silver has a maximum absorption peak at about 450nm, and the silver nitrate solution and the gynura bicolor extracting solution have no special absorption peak at the position.
FIG. 2 is a transmission electron microscope image (A) of the nano-silver solution and a distribution image (B) of the particle size of the nano-silver particles. It was determined that the nano silver solution prepared in this example had a silver ion particle size of 3 to 33nm and an average particle size of 13nm, as shown in fig. 2.
Example 2: EXAMPLE 1 evaluation of antibacterial Properties of the synthesized Nano-silver solution
(1) Strain culture: inoculating Escherichia coli, staphylococcus aureus and Pseudomonas aeruginosa in MH liquid culture medium, and culturing at 37 deg.C under 180r/min to logarithmic phase.
(2) The zone of inhibition test: respectively diluting escherichia coli, staphylococcus aureus and pseudomonas aeruginosa in logarithmic growth phase to 10 8 CFU/mL, 100. Mu.L each was spread evenly on MH solid medium. A plurality of sterilized filter paper sheets are respectively immersed in the nano-silver solution, the gynura bicolor extracting solution and the physiological saline which are synthesized in the embodiment 1, the filter paper sheets are stuck on an MH (metal hydride) plate loaded with escherichia coli, staphylococcus aureus and pseudomonas aeruginosa, the culture is carried out for 16 hours in an inverted way at the constant temperature of 37 ℃, the diameters of inhibition zones are respectively measured, and the average value is taken for 3 times. Each treatment was 3 replicates.
Table 1 shows the diameters of the zones of inhibition of the silver nitrate solution on Escherichia coli, staphylococcus aureus and Pseudomonas aeruginosa. Table 1 shows that, compared with normal saline and gynura bicolor extract, the obtained nano silver solution has high-efficiency antibacterial activity on escherichia coli, staphylococcus aureus and pseudomonas aeruginosa.
TABLE 1 diameter of the zone of inhibition of nanosilver solution to Escherichia coli, staphylococcus aureus and Pseudomonas aeruginosa
FIG. 3 is a diagram showing the results of the zone of inhibition test of silver nitrate solution to Escherichia coli.
FIG. 4 is a graph of the results of a test of the zone of inhibition of silver nitrate solution to Staphylococcus aureus.
FIG. 5 is a graph showing the results of a zone of inhibition test of silver nitrate solution on Pseudomonas aeruginosa.
The above description is only a preferred embodiment of the present invention and should not be taken as limiting the scope of the present invention, and the present invention is not limited thereto, and obvious modifications, equivalent substitutions, improvements, etc. within the technical spirit of the present invention should be included in the scope of the present invention.
Claims (6)
1. A method for preparing nano-silver based on gynura bicolor extract is characterized by comprising the following specific steps:
(1) Washing leaf part of gynura bicolor with water, drying, and pulverizing with a pulverizer to obtain gynura bicolor powder;
(2) According to mass-to-volume ratio =1g: adding the gynura bicolor powder into an ethanol water solution with the volume fraction of 75% in a proportion of 20-30 ml, carrying out hot reflux on the mixture at 80 ℃ for 2h, stirring continuously during the hot reflux, cooling the mixture to room temperature, carrying out ultrasonic treatment under an ice bath condition, and filtering to obtain a gynura bicolor ethanol extracting solution;
(3) Adding 10mmol/L AgNO 3 The solution and the gynura bicolor ethanol extract are mixed according to the volume ratio of 5:1, heating and stirring at 95 ℃ for reaction for 20min to prepare the nano silver solution.
2. The method of claim 1, wherein the washing with water of the leaf portion of the gynura bicolor is a washing with ultra pure water 3 times.
3. The method of claim 1, wherein the drying is drying at 65 ℃ of 1-2 d.
4. The method of claim 1, wherein the gynura bicolor powder is a gynura bicolor powder which is crushed by a crusher and passed through a 40-mesh sieve.
5. The method of claim 1, wherein the ultrasonic treatment under ice bath conditions is ultrasonic treatment under ice bath conditions with ultrasonic power of 300W for 30min for 2sec and 4 sec.
6. The method of claim 1, wherein said filtering to obtain an ethanol extract of gynura bicolor is performed with a 0.22 μ M filter.
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| CN107671305B (en) * | 2017-09-22 | 2019-05-10 | 鲁东大学 | The method for quickly preparing nano silver antibacterial agent using Ligustrum quihoui fruit extracting solution |
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