Method for preparing ion-conjugated material and metal coordination material thereof, and use thereof

Abstract

Disclosed in the present invention are an ion-conjugated material and a metal coordination material thereof, and the antibacterial use thereof, wherein 2,9-diformyl-1,10-phenanthroline and 1,2,3-triaminoguanidine hydrochloride are brought into a reaction to prepare an ion-conjugated material, and metal ions are coordinated on the ion-conjugated material to form a new metal coordination covalent organic framework material, which has an efficient antibacterial and sterilizing performance against Escherichia coli and Staphylococcus aureus, and a greatly improved sterilizing performance under light irradiation. Specifically, in practice, the sterilizing rate against Escherichia coli and Staphylococcus aureus can reach 99% or more; furthermore, under irradiation with an ultraviolet lamp, the sterilizing performance against high-concentration bacteria is greatly improved.

Description

Preparation method and application of ion conjugated material and metal coordination material thereof Technical Field

[0001] The present invention relates to the field of antibacterial polymer materials, and in particular to an ion conjugated material of a metal-coordinated covalent organic framework and a preparation method thereof, which has high antibacterial properties against Escherichia coli and Staphylococcus aureus, and specifically to a preparation method and application of an ion conjugated material and a metal coordination material thereof. Background Art

[0002] Bacterial infections have long been a threat to human health. For example, Escherichia coli infections can cause diarrhea, and in severe cases, dehydration and low blood pressure. Staphylococcus aureus infections, on the other hand, can lead to fever, vomiting, diarrhea, and, in severe cases, shock or even death. Antibiotics are a traditional treatment for bacterial infections, but as bacteria evolve and develop drug resistance, traditional antibiotics are becoming less effective. Therefore, the development of new antimicrobial materials is crucial. Summary of the Invention

[0003] In view of the need to develop antibacterial materials with better performance in the existing technology, the present invention discloses a class of ionic conjugate materials of metal coordination covalent organic frameworks and their preparation methods, which have high antibacterial properties against Escherichia coli and Staphylococcus aureus.

[0004] The cationic covalent organic framework used in the present invention is a type of ionic conjugated material with N in the ground state on the main chain. + ions, surrounded by stoichiometric amounts of Cl - ions, belonging to the second type of ion conjugated materials. After coordinating with metal ions, it forms a non-intrinsic fourth type of ion conjugated material. This ion conjugated property makes this material exhibit good chemical stability before and after metal coordination, which also allows this material to maintain a stable ionic state in water. The experimental results show that Zn 2+ / TGH + PD and Ag + / TGH + ·PD at 0.1 mg / mL for a bacterial solution concentration of 5×10 4 The sterilization rate of CFU / mL Escherichia coli and Staphylococcus aureus reached more than 99%, among which Ag + / TGH + PD has an outstanding antibacterial property against Escherichia coli. When 10 μg / mL is used, the concentration of the bacterial solution is 5×10 4 The sterilization rate of E. coli CFU / mL reached 99.99%. And under the ultraviolet light of 405 nm wavelength (50mW / cm 2) Under the condition of irradiation for 30 minutes, its bactericidal performance against higher concentrations of E. coli was greatly improved. 15μg / mL TGH + ·PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD and Ag + / TGH + ·PD for 1×10 7 The sterilization rates of E. coli at 100 CFU / mL were 63.54%, 92.02%, 70.87% and 99.99% respectively. + PD is exposed to UV light at 405 nm (50 mW / cm 2 ) Under the condition of irradiation for 30 minutes, for 10 6 The sterilization rate of CFU / mL reached 99.68%; 2μg / mL of Ag + / TGH + PD at 405 nm wavelength UV light (50 mW / cm 2 ) Under the condition of irradiation for 30 minutes, for 10 7 The sterilization rate of Escherichia coli CFU / mL reaches 99.99%.

[0005] Specifically, the present invention adopts the following technical solutions:

[0006] The present invention discloses the use of an ion-conjugated material or a metal-coordinated covalent organic framework material as or in the preparation of an antibacterial material. Preferably, the ion-conjugated material or the metal-coordinated covalent organic framework material is used as or in the preparation of a photocatalytic antibacterial material; further preferably, the photocatalysis is ultraviolet light catalysis, such as ultraviolet light catalysis at a wavelength of 405 nm.

[0007] The present invention discloses the above-mentioned ion conjugated material (TGH + The preparation method of PD comprises the following steps: reacting 2,9-diformyl-1,10-phenanthroline with 1,2,3-triaminoguanidine hydrochloride in a solvent to obtain the ion conjugated material (TGH + PD). Preferably, the molar ratio of 2,9-diformyl-1,10-phenanthroline to 1,2,3-triaminoguanidine hydrochloride is (1.2-2):1, preferably (1.4-1.8):1, for example including but not limited to 1.5:1; the reaction temperature can be 100-150° C., and the reaction time can be 0.5-2 hours; the solvent is a mixed solvent consisting of water and an organic solvent, preferably, the organic solvent can be dioxane.

[0008] The present invention discloses the above-mentioned metal coordinated covalent organic framework material, the raw materials of which include the above-prepared ion conjugated material and metal ions; preferably, the metal represented by the metal ions can include one or a combination of platinum, copper, zinc, gold, and silver.

[0009] The present invention discloses a method for preparing the above-mentioned metal-coordinated covalent organic framework material, comprising the following steps: reacting a metal salt with the ion-conjugated material in a solvent to obtain the metal-coordinated covalent organic framework material. The type of metal salt is selected based on the above-mentioned metal, specifically using conventional techniques, such as metal halides, metal sulfates, metal nitrates, etc., including but not limited to PtCl2, ZnCl2, AgNO3, etc. The molar amount of the metal ion in the metal salt is calculated according to the following formula:

[0010]

[0011] Among them, n 金属离子 is the molar amount of metal ions in the metal salt, m PD is the mass of 2,9-diformyl-1,10-phenanthroline, M PD is the molecular weight of 2,9-diformyl-1,10-phenanthroline, m TGH•Cl is the mass of 1,2,3-triaminoguanidine hydrochloride, m TGH + •PD is the mass of the ion conjugated material. + The molar amount of 2,9-diformyl-1,10-phenanthroline (PD) added during the PD reaction was the same.

[0012] When the metal salt is a platinum salt, the solvent is a mixed solution of ammonia water and water, and the reaction is carried out at room temperature for 5 to 20 hours to obtain the metal coordinated covalent organic framework material.

[0013] When the metal salt is zinc salt, the solvent is ethanol, and the reaction is carried out at 60-70° C. for 10-20 hours to obtain the metal coordinated covalent organic framework material.

[0014] When the metal salt is a silver salt, the solvent is water, and the reaction is carried out at room temperature for 8 to 15 hours to obtain the metal coordinated covalent organic framework material; preferably, the reaction is carried out in the dark.

[0015] The present invention discloses an antibacterial method, which utilizes the above-mentioned ion conjugated material or metal coordinated covalent organic framework material to perform antibacterial treatment.

[0016] In the present invention, the bacteria that have been tested so far include Escherichia coli and Staphylococcus aureus.

[0017] The present invention discloses a class of ion conjugate materials or metal coordination covalent organic framework materials with high antibacterial properties against Escherichia coli and Staphylococcus aureus, including Pt 2+ / TGH + PD, Zn 2+ / TGH + PD and Ag + / TGH + PD and its preparation method. The ion conjugated material or metal coordinated covalent organic framework material of the present invention has good antibacterial properties against Escherichia coli and Staphylococcus aureus, especially under the condition of relatively short-term ultraviolet light irradiation.

[0018] As an example, the prepared TGH + PD is dispersed in water, PtCl2 is dissolved in a mixture of ammonia and water, and Pt 2+ The solution was added dropwise to TGH + ·PD suspension, stirred, reacted at room temperature for 12 hours to obtain Pt 2+ / TGH + ·PD.

[0019] As an example, Zn 2+ / TGH + The preparation method of PD includes the following steps: ZnCl2, TGH + PD was dissolved in anhydrous ethanol and stirred at 70℃ for 12 hours to obtain Zn 2+ / TGH + ·PD.

[0020] As an example, Ag + / TGH + The preparation method of PD includes the following steps: + PD is dispersed in water, AgNO3 is dissolved in water, and Ag + The solution was added dropwise to TGH + ·PD suspension, stirred, reacted at room temperature for 12 hours to obtain Ag + / TGH + ·PD.

[0021] The present invention is a kind of metal ion (Pt 2+ / Zn 2+ / Ag +) is coordinated on the covalent organic framework to form a new type of ion conjugated material, namely the metal-coordinated covalent organic framework material, which has high antibacterial and bactericidal properties against Escherichia coli and Staphylococcus aureus, and the bactericidal properties are greatly enhanced under light. Specifically, the covalent organic framework TGH is prepared using triaminoguanidine hydrochloride and 2,9-diformyl-1,10-phenanthroline as raw materials and a mixed solution of 1,4-dioxane and water as solvent. + PD; then PtCl2, ZnCl2 or AgNO3 and TGH + PD reaction to prepare metal ions and TGH + The present invention selects Escherichia coli and Staphylococcus aureus as the sterilization test objects, and the concentration of bacteria in 1 mL is 5×10 4 CFU / mL solution, a certain amount of this antibacterial material was added and cultured at 37℃ in a dark environment for 24 hours. The final result was that Zn 2+ / TGH + PD and Ag + / TGH + ·PD at 0.1 mg / mL for a bacterial solution concentration of 5×10 4 The sterilization rate of CFU / mL Escherichia coli and Staphylococcus aureus reached more than 99%, among which Ag + / TGH + PD has an outstanding antibacterial property against Escherichia coli. When 10 μg / mL is used, the concentration of the bacterial solution is 5×10 4 The sterilization rate of E. coli CFU / mL reached 99.99%. And under the ultraviolet light of 405 nm wavelength (50mW / cm 2 ) Under the condition of irradiation for 30 minutes, its bactericidal performance against higher concentrations of E. coli was greatly improved. 15μg / mL TGH + PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD and Ag + / TGH + ·PD for 1×10 7 The sterilization rates of E. coli at 100 CFU / mL were 63.54%, 92.02%, 70.87% and 99.99% respectively. + PD is exposed to UV light at 405 nm (50 mW / cm 2 ) irradiation for 30 minutes for 10 6 The sterilization rate of CFU / mL reached 99.68%. 2μg / mL of Ag + / TGH +PD at 405 nm wavelength UV light (50 mW / cm 2 ) irradiation for 30 minutes for 10 7 The sterilization rate of Escherichia coli CFU / mL reaches 99.99%.

[0022] Compared with the prior art, the present invention using the above technical solution has the following advantages:

[0023] (1) The material disclosed in the present invention is easy to prepare and simple to operate;

[0024] (2) The material structure disclosed in the present invention is adjustable;

[0025] (3) The illumination time disclosed by the present invention is short and the operation is simple, that is, the sterilization rate can be greatly improved under shorter illumination conditions;

[0026] (4) The material disclosed in the present invention has a high sterilization rate against Escherichia coli and Staphylococcus aureus, and a low minimum antibacterial concentration;

[0027] (5) The antibacterial properties of the materials disclosed in the present invention against Escherichia coli are significantly improved under a relatively short period of ultraviolet light irradiation;

[0028] (6) The material disclosed in the present invention has a high sterilization rate against Escherichia coli under a relatively short period of ultraviolet light irradiation, and the minimum antibacterial concentration is extremely low. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 shows the ion conjugated material (TGH + PD), metal-coordinated covalent organic framework materials and their reaction diagrams;

[0030] Figure 2 shows the four materials (TGH + PD, TGH + PD / Pt, TGH + PD / Ag, TGH + PD / Zn) XRD pattern;

[0031] Figure 3 shows infrared images of four materials;

[0032] Figure 4 is a summary of the X-ray photoelectron spectra of the four materials;

[0033] FIG5 is a detailed X-ray photoelectric spectrum of C, N, Pt, Zn, and Ag elements in the four materials;

[0034] Figure 6 shows TGH + PD's MRI 13 C spectrum;

[0035] Figure 7 is a scanning electron microscope image of the four materials;

[0036] FIG8 is a UV-visible spectrum of four materials;

[0037] Figure 9 shows high concentration TGH + PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD, Ag + / TGH + PD pair 5×10 4 CFU / mL antibacterial properties against Escherichia coli and Staphylococcus aureus (at concentrations of 0.1, 0.5, and 2.5 mg / mL, respectively);

[0038] FIG10 is a bar graph showing the survival rates of E. coli of four materials at different concentrations;

[0039] FIG11 is a bar graph showing the survival rates of Staphylococcus aureus using four materials at different concentrations;

[0040] Figure 12 shows low concentration Zn 2+ / TGH + PD, Ag 2+ / TGH + PD pair 5×10 4 CFU / mL antibacterial performance against Escherichia coli and Staphylococcus aureus (concentrations of 10, 20, 40, and 80 μg / mL, respectively)

[0041] Figure 13 shows the low concentration of 15 μg / mL TGH + PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD, Ag + / TGH + PD at 405 nm wavelength, optical power 50 mW / cm 2 UV light for 30 minutes on 10 7 CFU / mL antibacterial performance test results of Escherichia coli;

[0042] Figure 14 shows the four materials at low concentration at a wavelength of 405 nm and a light power of 50 mW / cm 2 UV light irradiation for 30 minutes, the concentration is 10 7 Histogram of E. coli survival rate in CFU / mL;

[0043] Figure 15 shows TGH + PD at 405 nm wavelength, optical power 50 mW / cm 2For 30 minutes of ultraviolet light irradiation, the concentration of 10 6 Minimum antimicrobial concentration of CFU / mL Escherichia coli;

[0044] Figure 16 shows Ag + / TGH + PD at 405 nm wavelength, optical power 50 mW / cm 2 For 30 minutes of ultraviolet light irradiation, the concentration of 10 7 The lowest antibacterial concentration of CFU / mL Escherichia coli. DETAILED DESCRIPTION

[0045] The present invention belongs to the field of antibacterial materials, and specifically relates to a class of ionic conjugated materials of metal-coordinated covalent organic frameworks and their preparation methods. The materials have high antibacterial properties against Escherichia coli and Staphylococcus aureus, and their bactericidal properties against Escherichia coli are greatly improved under brief irradiation with 405 nm wavelength ultraviolet light.

[0046] Regulating the release rate of metal ions, improving biosafety, and addressing the poor antibacterial properties of low-concentration materials have always been challenges in the use of covalent organic frameworks (COFs) to load metal nanoparticles or coordinate with metal ions. Furthermore, many current COF materials used for antibacterial purposes still exhibit unsatisfactory performance under photocatalytic conditions. The present invention discloses a class of ion-conjugated materials based on metal-coordinated covalent organic frameworks, which have the advantages of a large specific surface area, a loose and porous structure (referring to a structure with numerous pores interconnected, non-interconnected, or partially interconnected, forming a porous and fluffy structure), good chemical stability, and ease of preparation. These materials effectively achieve highly effective antibacterial properties against Escherichia coli and Staphylococcus aureus, and can play an important role in many fields.

[0047] The present invention demonstrates the progress of the technical solution through antibacterial experiments.

[0048] The present invention has a concentration of 5×10 4 0.1, 0.5, and 2.5 mg of TGH were added to the Escherichia coli / Staphylococcus aureus culture solution containing 100 CFU / mL of TGH. + PD or Pt 2+ / TGH + PD or Zn 2+ / TGH + PD or Ag + / TGH + ·PD.

[0049] The present invention has a concentration of 5×10 4 10, 20, 40, and 80 μg of Zn were added to the Escherichia coli / Staphylococcus aureus culture solution containing 100 CFU / mL of Zn. 2+ / TGH + PD or Ag + / TGH + ·PD.

[0050] The present invention has a concentration of 5×10 4 TGH was added to the E. coli / S. aureus solution containing CFU / mL + PD or Pt 2+ / TGH + PD or Zn 2+ / TGH + PD or Ag + / TGH + ·PD.

[0051] The present invention has a concentration of 1×10 7 15 μg of TGH was added to the E. coli culture solution containing CFU / mL + PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD or Ag + / TGH + ·PD.

[0052] The present invention has a concentration of 1×10 7 TGH was added to the E. coli culture solution containing CFU / mL + PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD or Ag + / TGH + PD, using 405 nm UV light (50 mW / cm 2 ) for 30 minutes and then placed in an air atmosphere incubator at 37°C for 24 hours.

[0053] The bacterial dilution solution and the material dispersion solution were both sterile PBS solutions; and the co-culture time with the bacteria was 24 hours.

[0054] The present invention reacts 2,9-diformyl-1,10-phenanthroline with 1,2,3-triaminoguanidine hydrochloride in a solution to obtain the ion conjugated material (TGH + PD); reacting a metal salt with the ion conjugated material in a solution to obtain the metal coordinated covalent organic framework material. The synthesis steps are shown in Figure 1.

[0055] The raw materials used in this invention are all existing products, and the specific preparation methods and performance testing are conventional techniques. The Escherichia coli and Staphylococcus aureus used in this invention are from Wenzhou Weiqiong Microbiological Technology Co., Ltd., where the Escherichia coli is HC-16007 and the strain number is CMCC(B)44102; the Staphylococcus aureus is HC-16005 and the strain number is CMCC(B)26003.

[0056] The formula for calculating the sterilization rate is as follows:

[0057]

[0058] The colony count refers to the number of bacterial colonies on the LB medium after the culture is completed.

[0059] Example 1 Ionic Conjugated Covalent Organic Framework (TGH + Synthesis of PD)

[0060] TGH + The preparation method of PD is as follows: 106.2 mg (0.45 mmol) of 2,9-diformyl-1,10-phenanthroline and 42.4 mg (0.30 mmol) of 1,2,3-triaminoguanidine hydrochloride are dispersed in 2 mL of a mixture of 1,4-dioxane and water (v:v = 1:0.6). The mixture is placed in a 25 mL reactor and heated at 120°C for 1 hour. After heating, the mixture is filtered and washed three times with 1,4-dioxane and anhydrous ethanol. Finally, the resulting solid product is dried in a vacuum oven at 110°C for 12 hours to obtain the ionic conjugated covalent organic framework (COF) - TGH. + ·PD.

[0061] Example 2 Pt 2+ Coordination ion conjugated covalent organic framework (Pt 2+ / TGH + Synthesis of PD)

[0062] Take 100 mg TGH + PD was dispersed in 50 mL of water; 43 mg (0.24 mmol) of PtCl2 was dissolved in a mixture of 10 mL of ammonia and 40 mL of water. The PdCl2 solution was added dropwise to the TGH + ·PD suspension, stirred at room temperature for 12 hours. After the reaction, filtered, washed with water and PBS solution, and the obtained solid product was vacuum dried at 80 ° C for 3 hours to obtain the product Pt 2+ / TGH + ·PD.

[0063] Example 3 Zn2+ Coordination ion conjugated covalent organic framework (Zn 2+ / TGH + Synthesis of PD)

[0064] Take 100 mg TGH + PD and 33 mg (0.24 mmol) of ZnCl2 were dispersed in 25 mL of anhydrous ethanol and stirred at 70°C for 10 hours. After the reaction was completed, the mixture was filtered and washed with ethanol solution. The solid product was vacuum dried at 70°C for 3 hours to obtain the product Zn 2+ / TGH + ·PD.

[0065] Example 4 Ag + Coordination ion conjugated covalent organic framework (Ag / TGH + Synthesis of PD)

[0066] Take 100 mg TGH + PD, dispersed in 50 mL of water; 40.7 mg (0.24 mmol) of AgNO3 was dissolved in 50 mL of water. The AgNO3 solution was added dropwise to the TGH + The suspension of PD was stirred at room temperature for 12 hours, and the reaction was protected from light during the entire reaction process. After the reaction was completed, it was filtered, washed with water and PBS solution, and the obtained solid product was vacuum dried at 80 ° C for 3 hours to obtain the product Ag. + / TGH + ·PD.

[0067] Figure 2 shows the four materials (TGH + PD, TGH + PD / Pt, TGH + PD / Ag, TGH + Figure 3 is the infrared image of the four materials; Figure 4 is the X-ray photoelectron spectrum of the four materials; Figure 5 is the X-ray photoelectron spectrum of C, N, Pt, Zn, and Ag in the four materials; Figure 6 is the X-ray photoelectron spectrum of TGH + PD's MRI 13 C spectrum; Figure 7 is a scanning electron microscope image of the four materials; Figure 8 is a UV-visible spectrum of the four materials;

[0068] The identification data is as follows:

[0069] From NMR 13 The C spectrum shows that C atoms in different chemical environments correspond to different peaks. 13 C spectrum can prove TGH + Successful synthesis of PD.

[0070] From the XRD diagram, it can be seen that Pt, Zn, and Ag elements exist in the material in the form of ions; from the infrared spectrum, it can be seen that the aromatic CN bonds of the three metal-coordinated COFs have shifted and metal ion peaks are shown in the XPS diagram (X-ray photoelectron spectroscopy).

[0071] This demonstrates the successful coordination of metal ions.

[0072] Example 5 Four ionic conjugated metal coordinated covalent organic framework materials for 5×10 4 Antibacterial experiments on CFU concentration of Escherichia coli and Staphylococcus aureus

[0073] (1) Take 0.1 mL of revived Escherichia coli or Staphylococcus aureus liquid and measure the transmittance at 600 nm (OD 600 =1), and the bacterial solution concentration was 1×10 9 CFU / mL, a certain amount of bacterial solution was taken and mixed with sterile PBS solution to reduce the concentration of bacterial solution to 5×10 6 CFU / mL.

[0074] (2) Weigh the antibacterial material and add sterile PBS solution to prepare a suspension with a concentration of 1 mg / mL.

[0075] (3) Select a sterile 24-well plate and perform three replicates for each concentration of material to eliminate errors. For example, for a material with a concentration of 0.1 mg / mL, first add 0.89 mL of PBS solution to each well, then add 0.1 mL of material suspension, and finally add 0.01 mL of diluted bacterial solution. Ensure that the total amount of liquid in each well is 1 mL. At this time, the number of bacteria in each well is 5×10 4 After spotting, the plates were placed in an air-filled 37°C incubator to incubate the bacteria with the antimicrobial material for 24 hours. A control group for each concentration contained the same bacterial solution without the antimicrobial material.

[0076] (4) Remove the well plate after co-culture. Take 10 μl of liquid directly from the well of the 24-well plate and drop it on the LB medium in the culture dish. Spread it evenly with a spreader. Then, turn the coated culture dish upside down and place it in an incubator at 37°C under air atmosphere. Wait for the bacteria to grow for 24 hours.

[0077] (5) Take out the culture dish, take photos of the growth of the colonies, count the number of colonies, and calculate the sterilization rate.

[0078] Figure 9 shows high concentration TGH + PD, Pt 2+ / TGH + PD, Zn2+ / TGH + PD, Ag + / TGH + PD pair 5×10 4 The antibacterial performance of CFU / mL Escherichia coli and Staphylococcus aureus (concentrations are 0.1, 0.5, and 2.5 mg / mL respectively) is shown in Figure 10. + PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD, Ag + / TGH + The histograms corresponding to PD are arranged from left to right) The histograms of the survival rates of E. coli at different concentrations show that when all four materials are at high concentrations, E. coli is difficult to survive; Figure 11 shows the four materials (in the figure, TGH + PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD, Ag + / TGH + The histograms corresponding to PD are arranged from left to right. The results show that when all four materials are at high concentrations, it is difficult for Staphylococcus aureus to survive. Figure 12 shows the survival rate of Staphylococcus aureus at low concentrations of Zn 2+ / TGH + PD, Ag + / TGH + PD pair 5×10 4 Figure 2 shows the test results of the antibacterial properties of CFU / mL Escherichia coli and Staphylococcus aureus (at concentrations of 10, 20, 40, and 80 μg / mL, respectively).

[0079] The above test results show that the ionic conjugated covalent organic framework (TGH) prepared by the present invention + ·PD) and materials formed by metal coordination ion conjugated covalent organic frameworks have good antibacterial properties.

[0080] Example 6 Four ionic conjugated metal coordinated covalent organic framework materials 10 7 Photocatalytic antibacterial experiment on Escherichia coli with CFU / mL concentration

[0081] (1) Take 0.1 mL of revived Escherichia coli or Staphylococcus aureus liquid and measure the transmittance at 600 nm (OD 600 =1), and the bacterial solution concentration was 1×10 9 CFU / mL.

[0082] (2) Weigh the antibacterial material and add sterile PBS solution to prepare a 1 mg / mL suspension.

[0083] (3) Select a sterile 24-well plate and perform three replicates for each concentration of material to eliminate errors. For the 15 μg / mL concentration experimental group, first add 0.975 mL of PBS solution to each well, then add 0.015 mL of material suspension, and finally add 0.01 mL of diluted bacterial solution. Ensure that the total amount of liquid in each well is 1 mL. At this time, the number of bacteria in each well is 10 7 The control group for each concentration was the same concentration of bacterial solution without adding the antibacterial material.

[0084] (4) The wavelength used is 405 nm and the optical power is 50 mW / cm 2 UV light was placed on the well plate and illuminated for 30 minutes. Both the material group and the control group received illumination. After illumination, the well plate was inverted and placed in a 37°C incubator with air atmosphere to allow the bacteria and antimicrobial material to co-incubate for 24 hours.

[0085] (5) Take out the well plate after co-cultivation. Aspirate the bacterial solution in the well plate and dilute it to a bacterial concentration of about 10 5 CFU / mL of bacterial solution. Then, take 10 μl of the bacterial solution from this test tube and drop it onto the LB medium in a culture dish. Spread it evenly with a spreading stick. Then, invert the coated culture dish and place it in an air-filled incubator at 37°C to allow the bacteria to grow for 24 hours.

[0086] (6) Take out the culture dish, take photos of the growth of the colonies, count the number of colonies, and calculate the sterilization rate.

[0087] Figure 13 shows a low concentration of 15 μg / mL TGH + PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD, Ag + / TGH + PD at 405nm wavelength, optical power is 50mW / cm 2 UV light for 30 minutes on 10 7 Figure 14 shows the antibacterial performance test results of the four materials at low concentrations at a wavelength of 405 nm and a light power of 50 mW / cm 2 Histogram of E. coli survival rate under UV light irradiation for 30 minutes.

[0088] The above test results show that the ionic conjugated covalent organic framework (TGH) prepared by the present invention + ·PD) and materials formed by metal coordination ion conjugated covalent organic frameworks have good antibacterial properties.

[0089] Example 7 Ionic conjugated covalent organic framework material (TGH + PD) to 10 6 Experiment on the minimum antibacterial concentration of photocatalysis against Escherichia coli with CFU / mL concentration

[0090] (1) Take 0.1 mL of the revived Escherichia coli / Staphylococcus aureus solution and measure the transmittance at 600 nm (OD 600 =1), and the bacterial solution concentration was 1×10 9 CFU / mL.

[0091] (2) Weigh the antibacterial material and add sterile PBS solution to prepare a 1 mg / mL suspension.

[0092] (3) Select a sterile 24-well plate and perform three replicates for each concentration of material to eliminate errors. For the 2 μg / mL concentration experimental group, first add 0.988 mL of PBS solution to each well, then add 0.002 mL of material suspension, and finally add 0.01 mL of diluted bacterial solution. Ensure that the total amount of liquid in each well is 1 mL. At this time, the number of bacteria in each well is 10 6 The control group for each concentration was the same concentration of bacterial solution without adding the antibacterial material.

[0093] (4) The wavelength is 405 nm and the optical power is 50 mW / cm 2 UV light was placed on the well plate and illuminated for 30 minutes. Both the material group and the control group received illumination. After illumination, the well plate was inverted and placed in a 37°C incubator with air atmosphere to allow the bacteria and antimicrobial material to co-incubate for 24 hours.

[0094] (5) Take out the well plate after co-cultivation. Aspirate the bacterial solution in the well plate and dilute it to a bacterial concentration of about 10 5 CFU / mL of bacterial solution. Then, take 10 μl of the bacterial solution from this test tube and drop it onto the LB medium in a culture dish. Spread it evenly with a spreading stick. Then, invert the coated culture dish and place it in an air-filled incubator at 37°C to allow the bacteria to grow for 24 hours.

[0095] (6) Take out the culture dish, take photos of the growth of the colonies, count the number of colonies, and calculate the sterilization rate.

[0096] Figure 15 shows TGH + PD at 405 nm wavelength, optical power 50 mW / cm2 For 30 minutes of ultraviolet light irradiation, the concentration of 10 6 The lowest antibacterial concentration of CFU / mL Escherichia coli.

[0097] Example 8 Metal coordination ion conjugated covalent organic framework material (Ag + / TGH + PD) to 10 7 Experiment on the minimum antibacterial concentration of photocatalysis against Escherichia coli with CFU / mL concentration

[0098] (1) Take 0.1 mL of the revived E. coli / S. aureus solution and measure the transmittance at 600 nm (OD 600 =1), and the bacterial solution concentration was 1×10 9 CFU / mL.

[0099] (2) Weigh the antibacterial material and add sterile PBS solution to prepare a 1 mg / mL suspension.

[0100] (3) Select a sterile 24-well plate and perform three replicates for each concentration of material to eliminate errors. For the experimental group with a concentration of 2 μg / mL, first add 0.988 mL of PBS solution to each well, then add 0.002 mL of material suspension, and finally add 0.01 mL of diluted bacterial solution. Ensure that the total amount of liquid in each well is 1 mL. At this time, the number of bacteria in each well is 10 7 The control group for each concentration was the same concentration of bacterial solution without adding the antibacterial material.

[0101] (4) The wavelength used is 405 nm and the optical power is 50 mW / cm 2 UV light was placed on the well plate and illuminated for 30 minutes. Both the material group and the control group received illumination. After illumination, the well plate was inverted and placed in a 37°C incubator with air atmosphere to allow the bacteria and antimicrobial material to co-incubate for 24 hours.

[0102] (5) Take out the well plate after co-cultivation. Aspirate the bacterial solution in the well plate and dilute it to a bacterial concentration of about 10 5 CFU / mL of bacterial solution. Then, take 10 μl of the bacterial solution from this test tube and drop it onto the LB medium in a culture dish. Spread it evenly with a spreading stick. Then, invert the coated culture dish and place it in an air-filled incubator at 37°C to allow the bacteria to grow for 24 hours.

[0103] (6) Take out the culture dish, take photos of the growth of the colonies, count the number of colonies, and calculate the sterilization rate.

[0104] Figure 16 shows Ag + / TGH +PD at 405 nm wavelength, optical power 50 mW / cm 2 For 30 minutes of ultraviolet light irradiation, the concentration of 10 7 The lowest antibacterial concentration of CFU / mL Escherichia coli.

[0105] Example 9 Antibacterial material TGH of the present invention + PD and Ag + / TGH + Comparison of antibacterial performance of PD under photocatalytic conditions with existing technologies

[0106] Table 1 is the TGH of the present invention + PD and reported IBU@DhaTph-membrane, COF BDP / CD-S-3, Er-Co-S composite photocatalytic antibacterial material (Er / Co=1.5%mol) and Ag2TaNb7O 21 Comparison of the antibacterial performance of Ag in the present invention under suitable photocatalytic conditions. + / TGH + PD and reported Ag@DhaTph-COOH, COF BDP / CD-S-3-Ag, CFR@UiO-66(NH2)@Ag2S and Ag2TaNb 6.51 V 0.49 O 21 Comparison of antibacterial performance against Escherichia coli under suitable photocatalytic conditions.

[0107] Table 1 TGH of the present invention + Comparison of PD with existing technologies

[0108]

[0109] Table 2 Ag of the present invention + / TGH + Comparison of PD with existing technologies

[0110]

[0111] In summary, the present invention achieves high efficiency antibacterial activity against Escherichia coli and Staphylococcus aureus by preparing a class of ion-conjugated covalent organic frameworks and adding metal ion coordination. Moreover, the material of the present invention can effectively inhibit Escherichia coli and Staphylococcus aureus at low concentrations. The final result is that Zn 2+ / TGH + PD and Ag + / TGH + ·PD at 0.1 mg / mL for a bacterial solution concentration of 5×10 4The sterilization rate of CFU / mL Escherichia coli and Staphylococcus aureus reached more than 99%, among which Ag + / TGH + PD has an outstanding antibacterial property against Escherichia coli. When 10 μg / mL is used, the concentration of the bacterial solution is 5×10 4 The sterilization rate of CFU / mL Escherichia coli was 99.99%. 2 ) Under the condition of irradiation for 30 minutes, its bactericidal performance against higher concentrations of E. coli was greatly improved. 15μg / mL TGH + PD, Pt 2+ / TGH + PD, Zn 2+ / TGH + PD and Ag + / TGH + ·PD for 1×10 7 The sterilization rates of E. coli at 100 CFU / mL were 63.54%, 92.02%, 70.87% and 99.99% respectively. + PD at 405 nm wavelength UV light (50 mW / cm 2 ) irradiation for 30 minutes for 10 6 The sterilization rate of CFU / mL reached 99.68%. 2μg / mL of Ag + / TGH + PD at 405 nm wavelength UV light (50 mW / cm 2 ) irradiation for 30 minutes for 10 7 The sterilization rate of Escherichia coli CFU / mL reached 99.99%. Compared with other reported photocatalytic silver-containing covalent organic framework materials, the TGH of the present invention + PD and Ag + / TGH + PD is suitable for killing high concentrations of E. coli and has an extremely low minimum antibacterial concentration and a higher sterilization rate.

[0112] The ionic conjugate material of the present invention with high antibacterial properties against Escherichia coli and Staphylococcus aureus solves the current problem of poor antibacterial effect against Escherichia coli and Staphylococcus aureus at low material concentrations, as well as the problem of poor antibacterial effect against high-concentration Escherichia coli at low material concentrations under short-time illumination conditions.

[0113] The above embodiments are intended only to illustrate the technical concepts and features of the present invention. Their purpose is to enable those skilled in the art to understand the contents of the present invention and implement them accordingly. They are not intended to limit the scope of protection of the present invention. Any equivalent changes or modifications made in accordance with the spirit of the present invention are intended to be covered by the scope of protection of the present invention.

[0114] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

Claims

1. An ion conjugated material, characterized in that: The ionic conjugate material is prepared by reacting 2,9-diformyl-1,10-phenanthroline with 1,2,3-triaminoguanidine hydrochloride.

2. The ion conjugated material according to claim 1, characterized in that The ion conjugated material has a porous structure.

3. The ion conjugated material according to claim 1, characterized in that The ion conjugated material has a loose porous structure.

4. The ion conjugated material according to claim 1, characterized in that The main chain of the ion conjugated material has N in the ground state. + ions, surrounded by stoichiometric amounts of Cl - ions, and the ion conjugated material can maintain an ionic state in water.

5. A method for preparing the ion conjugated material according to any one of claims 1 to 4, characterized in that: The preparation method comprises: 2,9-diformyl-1,10-phenanthroline is reacted with 1,2,3-triaminoguanidine hydrochloride in water and an organic solvent to produce the ion conjugated material.

6. The method for preparing an ion conjugated material according to claim 5, characterized in that: The organic solvent includes dioxane.

7. The method for preparing an ion conjugated material according to claim 5, wherein: The reaction is controlled to be carried out at 100-150° C.; and / or the reaction time is controlled to be 0.5-2 h.

8. The method for preparing an ion conjugated material according to claim 5, wherein: The molar ratio of the 2,9-diformyl-1,10-phenanthroline to the 1,2,3-triaminoguanidine hydrochloride is 1.2-2:

1.

9. The method for preparing an ion conjugated material according to claim 8, characterized in that: The molar ratio of the 2,9-diformyl-1,10-phenanthroline to the 1,2,3-triaminoguanidine hydrochloride is 1.4-1.8:

1.

10. The antibacterial material according to claim 5, characterized in that: The embodiment of preparing the ion conjugated material includes: 2,9-diformyl-1,10-phenanthroline and 1,2,3-triaminoguanidine hydrochloride are dispersed in a mixed solvent consisting of water and an organic solvent respectively, reacted under heating conditions, and after the reaction is completed, filtered, washed, and vacuum dried to obtain the ionic conjugated material.

11. A metal coordinated covalent organic framework material, characterized in that: The metal coordinated covalent organic framework material is prepared by reacting an ion conjugated material with metal ions, and the ion conjugated material is prepared by reacting 2,9-diformyl-1,10-phenanthroline with 1,2,3-triaminoguanidine hydrochloride.

12. The metal coordinated covalent organic framework material according to claim 11, characterized in that: The metal coordination covalent organic framework material has a porous structure.

13. The metal coordinated covalent organic framework material according to claim 11, characterized in that: The metal coordinated covalent organic framework material has a loose porous structure.

14. The metal coordinated covalent organic framework material according to claim 11, characterized in that: The main chain of the metal coordinated covalent organic framework material has N in the ground state. + ions, surrounded by stoichiometric amounts of Cl - ions, and the metal coordinated covalent organic framework material can maintain an ionic state in water.

15. The metal coordinated covalent organic framework material according to claim 11, characterized in that: The metal ions include one or more combinations selected from platinum ions, copper ions, zinc ions, gold ions, and silver ions.

16. The metal coordinated covalent organic framework material according to claim 11, characterized in that The metal ions are provided by adding metal salts.

17. The metal coordinated covalent organic framework material according to claim 16, characterized in that: The metal salt comprises a combination of one or more selected from metal halogen salts, metal sulfates and metal nitrates.

18. The metal coordinated covalent organic framework material according to claim 16, characterized in that The metal salt is a combination of one or more selected from PtCl2, ZnCl2, and AgNO3.

19. A method for preparing the metal coordinated covalent organic framework material according to any one of claims 11 to 18, characterized in that: The preparation method of the metal coordinated covalent organic framework material comprises: (1) reacting 2,9-diformyl-1,10-phenanthroline with 1,2,3-triaminoguanidine hydrochloride in water and an organic solvent to generate an ion conjugated material; (2) Then, the ionic conjugated material is reacted with a metal salt in a solvent to obtain the metal coordinated covalent organic framework material.

20. The method for preparing a metal coordinated covalent organic framework material according to claim 19, characterized in that: The organic solvent includes dioxane; and / or the molar ratio of the 2,9-diformyl-1,10-phenanthroline to the 1,2,3-triaminoguanidine hydrochloride is 1.2-2:

1.

21. The method for preparing a metal coordinated covalent organic framework material according to claim 19, characterized in that: The reaction of 2,9-diformyl-1,10-phenanthroline with 1,2,3-triaminoguanidine hydrochloride is carried out at 100-150° C.; and / or the reaction of 2,9-diformyl-1,10-phenanthroline with 1,2,3-triaminoguanidine hydrochloride is carried out for 0.5-2 h.

22. The method for preparing a metal coordinated covalent organic framework material according to claim 19, characterized in that: The molar ratio of the 2,9-diformyl-1,10-phenanthroline to the 1,2,3-triaminoguanidine hydrochloride is 1.4-1.8:

1.

23. The method for preparing a metal coordinated covalent organic framework material according to claim 19, characterized in that: When the metal salt is a platinum salt, the reaction between the ionic conjugate material and the metal salt is carried out in a mixed solution of ammonia and water; preferably, the reaction is controlled to react at room temperature for 5 to 20 hours; When the metal salt is a zinc salt, the reaction between the ionic conjugate material and the metal salt is carried out in ethanol; preferably, the reaction is controlled at 60-70° C. for 10-20 hours; When the metal salt is a silver salt, the reaction of the ionic conjugated material and the metal salt is carried out in water; preferably, the reaction is controlled to react at room temperature for 8 to 15 hours, and more preferably, the reaction is carried out in the dark.

24. The method for preparing a metal coordinated covalent organic framework material according to claim 19, wherein: The molar amount of the metal ion in the metal salt is calculated according to the following formula: ； Among them, n 金属离子 is the molar amount of metal ions in the metal salt, m PD is the mass of 2,9-diformyl-1,10-phenanthroline, M PD is the molecular weight of 2,9-diformyl-1,10-phenanthroline, m TGH•Cl is the mass of 1,2,3-triaminoguanidine hydrochloride, m TGH + •PD is the mass of the ion-conjugated material.

25. Use of the ionic conjugated material according to any one of claims 1 to 4, or the ionic conjugated material prepared by the preparation method according to any one of claims 5 to 10, or the metal coordination covalent organic framework material according to any one of claims 11 to 18, or the metal coordination covalent organic framework material prepared by the preparation method according to any one of claims 19 to 24 as or in the preparation of an antibacterial material.

26. The use according to claim 25, characterized in that The antibacterial treatment refers to antibacterial treatment of Escherichia coli and / or Staphylococcus aureus; and / or the antibacterial treatment is performed under light, and the light used in the light treatment includes ultraviolet light.

27. The use according to claim 25, characterized in that The antibacterial material is a photocatalytic antibacterial material or a non-photocatalytic antibacterial material.

28. An antibacterial method, characterized in that: 2,9-diformyl-1,10-phenanthroline is reacted with 1,2,3-triaminoguanidine hydrochloride in a solvent to obtain an ion conjugated material; a metal salt is reacted with the ion conjugated material in a solvent to obtain a metal coordinated covalent organic framework material; and the ion conjugated material or the metal coordinated covalent organic framework material is used for antibacterial treatment to achieve antibacterial effect.

29. The antibacterial method according to claim 28, characterized in that: The antibacterial treatment is a photocatalytic antibacterial treatment or a non-photocatalytic antibacterial treatment.

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