Composite cotton cloth loaded with photosensitizer sodium copper chlorophyllin and preparation method and application thereof

Abstract

The present application relates to the composite cotton cloth loaded with sodium copper chlorophyllin photosensitizer and its preparation method and application, belong to the technical field of antibacterial material.The preparation method of the present application comprises the following steps: soaking cotton cloth in lye and heating treatment to obtain alkalized cotton cloth;placing the obtained alkalized cotton cloth in an organic solvent containing tricyanogen chloride and heating to obtain tricyanogen chloride grafted cotton cloth;immersing the obtained tricyanogen chloride grafted cotton cloth in an aqueous solution of sodium copper chlorophyllin and heating to react, after the reaction is completed, washing, drying to obtain the light-powered antibacterial cotton cloth loaded with sodium copper chlorophyllin.The light-powered antibacterial cotton cloth prepared by the present application has the characteristics of high dyeing rate, high active sterilization efficiency, broad-spectrum sterilization, low price, safety to human body and environment, etc., and can be used in daily life and has the prospect of large-scale production.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of antibacterial materials, in particular to a composite cotton cloth loaded with sodium copper chlorophyllin and a preparation method and application thereof. BACKGROUND

[0002] Broad-spectrum antibacterial materials can effectively block the contact transmission of pathogenic microorganisms in public places, meeting the major needs of the current public health system to improve the prevention and control capabilities of infectious diseases. Among them, the photodynamic antibacterial material converts the energy of visible light into chemical energy through the interaction of photosensitizers embedded in the material and oxygen molecules, generating active oxygen with inactivation of microorganisms. This bactericidal process has the advantages of non-specificity, high efficiency, safety, etc. In recent years, the development of photodynamic antibacterial materials has also made considerable progress, however, the high price of commercial photosensitizers, and the potential threat of photosensitizers released from the material to the environment and human body, hinder the large-scale production and market promotion of photodynamic antibacterial materials. Exploring efficient and low-cost, safe to human body and environment photosensitizers, and based on this to prepare scalable antibacterial products, has become a key step for photodynamic antibacterial materials to successfully scale up and create positive economic benefits.

[0003] Currently, a large number of efficient photodynamic antibacterial materials have been designed, prepared and studied, in which the loaded photosensitizer can be roughly divided into four pyrrole macrocycle (porphyrin), synthetic dyes, nanoparticles and natural products four categories. With the four pyrrole macrocycle structure of the photosensitizer (porphyrin) represented by Photofrin@ approved by the US Food and Drug Administration (FDA) for clinical use, the theory and clinical research in the field of photodynamic have been very mature, but its relatively complex molecular structure determines that the synthesis of this compound is tedious, and the price of the finished product is high, which greatly limits the use of this photosensitizer in the photodynamic antibacterial material which aims to scale production and application in daily life. Some synthetic dyes with reasonable prices, represented by methylene blue (MB) and rose bengal (RB), are also widely used in the preparation of photodynamic materials. However, the safety of these chemical reagents to the human body, especially the side effects of high doses after enrichment, has caused people's concern. Nanoparticles with photosensitive reactions, such as TiO2 and carbon quantum dots, are also used as photosensitizers to prepare a series of photodynamic antibacterial materials. However, like nanometal antibacterial materials, this material has the risk of nanoparticle enrichment in the environment and organisms, which may cause potential harm to the human body and the environment. Through extraction, purification and other semi-synthetic means, compounds with photosensitive effects obtained from natural plants and animals are most likely to meet the requirements of low price and biological safety. At present, researches on natural products as photosensitizers applied in photodynamic antibacterial materials have made certain progress: Wang Qingqing's research group of Jiangnan University successfully loaded the hyporellin produced by the mycelium secondary metabolic synthesis pathway of the Hyporellaceae strain such as bamboo red fungus and bamboo yellow fungus on the bacterial cellulose through trichloroamine as a crosslinking agent, and the prepared film had a bactericidal efficiency of more than 99.5% on gram-positive Staphylococcus aureus under light conditions (30 minutes; 420-780 nm; 65±5 mW / cm 2 ). Gram-negative Escherichia coli showed strong resistance to the material, and needed to be treated with additional potassium iodide to extend the life cycle of the active substance, and under the same light conditions, the bactericidal efficiency reached 99.1%. Adriana Manso's research group at the University of British Columbia in Canada successfully loaded riboflavin, which is widely present in yeast, animal liver, eggs and milk, into a composite resin used for oral treatment, and under the conventional light conditions of dentistry (1 min; blue light; 1.3 J / cm 2), Gram-positive anaerobic Streptococcus exhibits strong resistance to materials. The research group of Lin Shaoling of Fujian Agriculture and Forestry University prepared a degradable antibacterial film using curcumin compound extracted from rhizomes of Zingiber officinale as a photosensitizer. The bactericidal efficiency of the antibacterial film on Staphylococcus aureus is about 90% under light conditions (30 min; blue light LED), and the bactericidal efficiency on Escherichia coli is about 70%. Although these materials have the advantages of biological safety and reasonable price, they generally fail to exhibit high inactivation efficiency of pathogenic microorganisms, especially individual microorganism species exhibit strong resistance. According to a literature of Roca team of Spanish National Research Council (CSIC), 2.5 mM (about 1.8 mg / L) of sodium copper chlorophyllin (Chl) solution can kill 99.9999% of Staphylococcus aureus under 1 hour of visible light irradiation. Sodium copper chlorophyllin is prepared by extracting chlorophyll and its derivatives from plants rich in chlorophyll through high-temperature extraction and the like, and then adding a divalent copper salt as a stabilizer. Because the raw material is easy to obtain, the preparation process is relatively simple, and the price is stable at about 1 yuan / gram. At the same time, Chl has a long history as a food additive (E140i) that has been widely used in the food industry. As early as 2002, FDA approved the use of Chl in food / drugs, and the safe daily intake of each adult is about 300 mg, which is much higher than the effective bactericidal value of Chl mentioned above. In view of the above data, the bactericidal efficiency and biological safety of sodium copper chlorophyllin as a photosensitizer are both well guaranteed, and it is highly possible to achieve the research goal of constructing an efficient, safe and broad-spectrum photodynamic antibacterial material. Although it has a promising future, the research on preparing photodynamic antibacterial materials using Chl as a photosensitizer is still a blank field, and there are many challenges. SUMMARY

[0004] To solve the above technical problems, the application provides a sodium copper chlorophyllin-loaded photodynamic antibacterial cotton cloth and a preparation method thereof. The prepared photodynamic antibacterial cotton cloth has high active bactericidal efficiency, broad-spectrum bactericidal effect, low price, safety to human body and environment, can be used in daily life and has a large-scale prospect. Moreover, the application successfully verifies that the inactivation efficiency of the photodynamic antibacterial cotton cloth on vancomycin-resistant Enterococcus is 99.998% under light conditions (30 minutes; 420-780 nm; 65±5 mW / cm 2 );the inactivation efficiency of the photodynamic antibacterial cotton cloth on methicillin-resistant Staphylococcus aureus is 99.994%;and the inactivation efficiency of the photodynamic antibacterial cotton cloth on feline calicivirus is 99.8%.

[0005] The application is implemented by the following technical solutions:

[0006] The first object of the present application is to provide a preparation method of a sodium copper chlorophyllin loaded composite cotton cloth, comprising the following steps:

[0007] (1) soaking cotton cloth in an alkali solution for heat treatment to obtain an alkalized cotton cloth;

[0008] (2) placing the alkalized cotton cloth obtained in step (1) in an organic solvent containing tricyanogen chloride for heat reaction to obtain a tricyanogen chloride grafted cotton cloth;

[0009] (3) immersing the tricyanogen chloride grafted cotton cloth obtained in step (2) in an aqueous solution of sodium copper chlorophyllin for heat reaction, and after the reaction is completed, washing and drying to obtain the sodium copper chlorophyllin loaded composite cotton cloth.

[0010] In an embodiment of the present application, in step (1), the alkali in the alkali solution is selected from sodium hydroxide and / or potassium hydroxide.

[0011] In an embodiment of the present application, in step (1), the heat treatment is performed at 10-40℃ for 0.5-6h.

[0012] In an embodiment of the present application, in step (2), the organic solvent is selected from one or more of tetrahydrofuran, acetone, acetonitrile, ethanol, acetic acid and chloroform.

[0013] In an embodiment of the present application, in step (2), the heat reaction is performed at 25-60℃ for 6-24h.

[0014] In an embodiment of the present application, in step (3), the molar ratio of tricyanogen chloride to sodium copper chlorophyllin is 1:1-6:1.

[0015] In an embodiment of the present application, in step (3), the heat is performed at 25-60℃ for 6-24h.

[0016] The second object of the present application is to provide the sodium copper chlorophyllin loaded composite cotton cloth obtained by the preparation method.

[0017] In an embodiment of the present application, the content of sodium copper chlorophyllin in the composite cotton cloth is 0.01wt%-0.1wt%.

[0018] The third object of the present application is to provide the application of the composite cotton cloth in antibiosis.

[0019] The fourth object of the present application is to provide the application of the composite cotton cloth in antiviral.

[0020] In an embodiment of the present application, the dyeing rate of the composite cotton cloth is 20%-60%.

[0021] The preparation of Chl-cotton can be roughly divided into three steps Figure 1 The first step is to place pure cotton in NaOH aqueous solution for 2 hours, then react the ionized cotton after soaking with a solution of tetrahydrofuran (THF) containing trichloro-1,3,5-triazine (TCT) at 40°C for 12 hours. The last step is to wash the cotton after the reaction is completed with THF and deionized water (DI water) to wash away the free TCT, and then react with an aqueous solution containing Chl (the molar ratio of TCT to Chl is 4:1) at 40°C for another 12 hours, and then repeatedly washed with DI until the washing liquid is transparent. The sample is taken out and dried, and stored in the dark to be used.

[0022] The above technical solutions of the present application have the following advantages compared with the prior art:

[0023] The present application provides a cotton cloth with photodynamic antibacterial properties loaded with copper sodium salt of chlorophyll as a photosensitizer. The photodynamic antibacterial cotton cloth can greatly reduce the cost of photodynamic antibacterial materials and solve the potential threat of free photosensitizer to the human body and the environment.

[0024] The present application provides a cotton cloth dyeing method for chemically grafting copper sodium salt of chlorophyll with trichloro-1,3,5-triazine. The method has high dyeing rate, can actively kill bacteria, has high broad-spectrum bactericidal efficiency, is safe to the human body and the environment, can be used in daily life, and has large-scale prospects. BRIEF DESCRIPTION OF DRAWINGS

[0025] In order to make the content of the present application more easily understood, the present application will be further described in detail below according to specific embodiments of the present application and in conjunction with the drawings, in which

[0026] Figure 1 is the synthesis route of the copper sodium salt of chlorophyll cotton cloth in the present application;

[0027] Figure 2 is the bacterial detection flowchart of the composite cotton cloth loaded with copper sodium salt of chlorophyll in the present application. DETAILED DESCRIPTION

[0028] The present application will be further described below in conjunction with the drawings and specific embodiments, so that those skilled in the art can better understand the present application and implement it, but the embodiments are not limiting to the present application.

[0029] Example 1

[0030] The embodiment provides a preparation method of composite cotton cloth loaded with sodium copper chlorophyllin, and specifically comprises the following steps.

[0031] (1) 15cm*15cm (about 5g) of cotton cloth is separately soaked in 100mL of 0.5M NaOH solution, and then heated at 40°C for 2h to obtain alkali-treated cotton cloth.

[0032] (2) The alkali-treated cotton cloth is placed in 200mL of 5mM tricyanopyridine tetrahydrofuran solution, heated at 40°C for 12h, and then washed with tetrahydrofuran and distilled water to remove free TCT, so as to obtain tricyanopyridine grafted cotton cloth.

[0033] (3) The tricyanopyridine grafted cotton cloth is then placed in 200mL of 1.25mM sodium copper chlorophyllin aqueous solution, and heated at 40°C for 12h (the molar ratio of TCT to Chl is 4:1) to continue the reaction, after the reaction is completed, the sample is washed with a large amount of clean water until the washing liquid is transparent, and then the sample is placed in a 40°C oven for drying, so as to obtain composite cotton cloth loaded with sodium copper chlorophyllin, which is placed in a cool place for standby.

[0034] Example 2

[0035] The embodiment provides a preparation method of composite cotton cloth loaded with sodium copper chlorophyllin, and specifically comprises the following steps.

[0036] (1) 15cm*15cm (about 5g) of cotton cloth is separately soaked in 100mL of 0.5M NaOH solution, and then heated at 40°C for 2h to obtain alkali-treated cotton cloth.

[0037] (2) The alkali-treated cotton cloth is placed in 200mL of 5mM tricyanopyridine tetrahydrofuran solution, heated at 40°C for 12h, and then washed with tetrahydrofuran and distilled water to remove free TCT, so as to obtain tricyanopyridine grafted cotton cloth.

[0038] (3) The tricyanopyridine grafted cotton cloth is then placed in 200mL of 0.25mM sodium copper chlorophyllin aqueous solution, and heated at 40°C for 12h (the molar ratio of TCT to Chl is 4:1) to continue the reaction, after the reaction is completed, the sample is washed with a large amount of clean water until the washing liquid is transparent, and then the sample is placed in a 40°C oven for drying, so as to obtain composite cotton cloth loaded with sodium copper chlorophyllin, which is placed in a cool place for standby.

[0039] Example 3

[0040] The embodiment provides a preparation method of composite cotton cloth loaded with sodium copper chlorophyllin, and specifically comprises the following steps.

[0041] (1) 15 cm x 15 cm (~5 g) of cotton cloth was separately immersed in 100 mL of 0.5 M NaOH solution, and then heated at a temperature of 40 °C for 2 h to obtain the alkali-treated cotton cloth.

[0042] (2) The alkali-treated cotton cloth was placed in 200 mL of 5 mM tricyanochloro triazine in tetrahydrofuran solution, and heated at a temperature of 40 °C for 12 h. The cotton cloth after the reaction was completed was washed with tetrahydrofuran and distilled water to wash away the free TCT, to obtain the tricyanochloro triazine grafted cotton cloth.

[0043] (3) The tricyanochloro triazine grafted cotton cloth was then placed in 200 mL of 2 mM copper sodium salt of chlorophyll in aqueous solution, and the reaction was continued at a temperature of 40 °C for 12 h (molar ratio of TCT to Chl was 4:1). After the reaction was completed, the sample was washed with a large amount of clean water until the washing liquid was transparent; then the sample was placed in an oven at 40 °C for drying, to obtain the composite cotton cloth loaded with copper sodium salt of chlorophyll, which was stored in a cool place for use.

[0044] Test Example

[0045] The composite cotton cloth loaded with copper sodium salt of chlorophyll prepared in Example 1 was tested by colony counting method. The composite cotton cloth loaded with copper sodium salt of chlorophyll prepared in Example 1 was incubated with the test bacteria overnight under light conditions (30 minutes; 420-780 nm; 65 ± 5 mW / cm 2 ). The number of colonies visible on the agar plate was determined by colony counting, and the survival rate and inactivation rate were determined by the ratio of CFU / mL of the light plate to the corresponding dark control without material. (The specific test method is shown in Figure 2

[0046] From the above test results, it can be seen that the composite cotton cloth loaded with copper sodium salt of chlorophyll obtained in Example 1 has a killing efficiency of 99.998% on vancomycin-resistant Enterococcus faecalis, and an inactivation efficiency of 99.994% on methicillin-resistant Staphylococcus aureus. Using a similar method, the feline calicivirus was detected, and the composite cotton cloth loaded with copper sodium salt of chlorophyll had an inactivation efficiency of 99.8% on feline calicivirus.

[0047] The content of copper sodium salt of chlorophyll in the composite cotton cloth loaded with copper sodium salt of chlorophyll prepared in Example was determined by inductively coupled plasma spectrometer (ICP) to be 0.01 wt% to 0.1 wt%.

[0048] ​Obviously, the above embodiments are merely example for clearly illustrating, and are not limitation to the embodiments. For ordinary skilled in the art, other different forms of changes or variations can be made on the basis of the above description. Here, all the embodiments need not and can not be exhausted. The obvious changes or variations derived therefrom are still within the scope of the present invention.

Claims

1. A method for preparing a composite cotton cloth loaded with sodium copper chlorophyllin, characterized by, The method comprises the following steps: (1) soaking cotton cloth in an alkali solution and heating to obtain alkali-treated cotton cloth; (2) placing the alkali-treated cotton cloth obtained in step (1) in an organic solvent containing cyanuric chloride and heating to obtain cyanuric chloride-grafted cotton cloth; (3) immersing the cyanuric chloride-grafted cotton cloth obtained in step (2) in an aqueous solution of copper sodium salt of chlorophyll and heating to react, and after the reaction, washing, drying to obtain the composite cotton cloth loaded with copper sodium salt of chlorophyll; In step (1), the heating treatment is performed at 10-40°C for 0.5-6 h; In step (2), the heating reaction is performed at 25-60°C for 6-24 h; In step (3), the molar ratio of cyanuric chloride in the cyanuric chloride-grafted cotton cloth to copper sodium salt of chlorophyll is 1:1-6:

1.

2. The production method according to claim 1, characterized by, In step (1), the alkali in the alkali solution is selected from sodium hydroxide and / or potassium hydroxide.

3. The preparation method according to claim 1, characterized in that, In step (2), the organic solvent is selected from one or more of tetrahydrofuran, acetone, acetonitrile, ethanol, acetic acid and chloroform.

4. The composite cotton cloth loaded with copper sodium salt of chlorophyll obtained by the preparation method of any one of claims 1-3.

5. The composite cloth according to claim 4, wherein, The content of copper sodium salt of chlorophyll in the composite cotton cloth is 0.01-0.1 wt%.

6. The application of the composite cotton cloth of claim 5 in antibacterial.

7. The application of the composite cotton cloth of claim 5 in antiviral.

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