A bacteriostatic solution containing copper and a method for preparing the same

Abstract

This invention relates to the field of microbial detection and antibacterial technology, and particularly to a method for preparing a copper-containing antibacterial solution, comprising the following steps: mixing a copper-containing component with an antibacterial enhancing component to obtain a copper-containing antibacterial solution; wherein the antibacterial enhancing component is selected from at least one of tea polyphenols, amino acid compounds, and hydroxycarboxylic acid salts. The method for preparing the copper-containing antibacterial solution provided by this invention achieves a significant enhancement of the antibacterial effect by constructing a stable compound system with the antibacterial enhancing component and the copper-containing component. Simultaneously, it reduces the amount of copper ions used, improving both the antibacterial effect and safety.

Description

Technical Field

[0001] This invention relates to the field of microbial detection and antibacterial technology, and in particular to a copper-containing antibacterial solution and its preparation method. Background Technology

[0002] Copper ions, as a broad-spectrum antibacterial agent, are widely used in various fields such as medicine, food, materials, and water treatment due to their advantages such as broad antibacterial range, low likelihood of drug resistance, and environmental friendliness. In existing technologies, the antibacterial effect of copper-containing solutions mainly relies on the action of copper ions, but this has significant limitations: firstly, high concentrations of copper ions are toxic to human cells and the environment, limiting their safe application in food, medical, and other fields; secondly, low concentrations of copper ions have limited antibacterial effects, making it difficult to meet the high requirements of antibacterial scenarios. Therefore, existing copper-containing antibacterial solutions struggle to balance antibacterial properties and safety, thus limiting their application. Summary of the Invention

[0003] To address the problem that existing copper-containing antibacterial solutions struggle to balance antibacterial efficacy and safety, this invention provides a method for preparing a copper-containing antibacterial solution. This method combines an antibacterial enhancement component with a copper-containing component to achieve a synergistic antibacterial effect of "low-concentration copper ions + antibacterial enhancement component," reducing the amount of copper ions used and decreasing toxicity. While improving the antibacterial effect, it also enhances safety, thus solving the problem that existing copper-containing antibacterial solutions struggle to balance antibacterial efficacy and safety.

[0004] The technical solution adopted by this invention to solve its technical problem is: A method for preparing a copper-containing antibacterial solution includes the following steps: mixing a copper-containing component with an antibacterial enhancing component to obtain a copper-containing antibacterial solution; wherein the antibacterial enhancing component is selected from at least one of tea polyphenols, amino acid compounds, and hydroxycarboxylic acid salt compounds.

[0005] Optionally, the mass ratio of the copper-containing component to the antibacterial enhancement component is 1:(5-10).

[0006] Optionally, the amino acid compound is lysine.

[0007] Optionally, the hydroxycarboxylic acid salt compound is potassium citrate.

[0008] Optionally, the antibacterial enhancing components include tea polyphenols, amino acid compounds, and hydroxycarboxylic acid salt compounds.

[0009] Optionally, mixing the copper-containing component with the antibacterial enhancing component includes: mixing the copper-containing component with a hydroxycarboxylate compound, and then sequentially adding an amino acid compound and tea polyphenols to obtain the copper-containing antibacterial solution.

[0010] Optionally, the mass ratio of the hydroxycarboxylic acid salt compound, the amino acid compound, and the tea polyphenol is 1:1:1.

[0011] Optionally, the copper-containing component includes copper gluconate.

[0012] Another object of the present invention is to provide a copper-containing antibacterial solution, which is prepared by the method described above for preparing a copper-containing antibacterial solution.

[0013] The beneficial effects of this invention are: The method for preparing a copper-containing antibacterial solution provided by the present invention achieves a significant enhancement of the antibacterial effect by constructing a stable compound system with antibacterial enhancement components and copper-containing components. At the same time, it can reduce the amount of copper ions used, thereby improving safety while enhancing the antibacterial effect. Detailed Implementation

[0014] The present invention will now be described in further detail. The embodiments described below are exemplary and intended to explain the present invention, and should not be construed as limiting the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0015] To address the problem that existing copper-containing antibacterial solutions cannot simultaneously achieve both antibacterial properties and safety, this invention provides a method for preparing a copper-containing antibacterial solution. The method includes the following steps: mixing a copper-containing component with an antibacterial enhancing component to obtain a copper-containing antibacterial solution; wherein the antibacterial enhancing component is selected from at least one of tea polyphenols, amino acid compounds, and hydroxycarboxylic acid salt compounds.

[0016] By combining antibacterial enhancement components with copper-containing components, a synergistic antibacterial effect of "low concentration of copper ions + antibacterial enhancement components" is achieved, reducing the amount of copper ions used and decreasing toxicity.

[0017] Specifically, the catechol / gallic acid group of tea polyphenols can interact with... The formation of bidentate chelates stabilizes the structure and alters the electron cloud distribution, thereby disrupting bacterial membranes, triggering ROS bursts, and degrading nucleic acids. When tea polyphenols and copper ions work synergistically, they can disrupt the integrity of bacterial cell membranes and enhance the permeability of copper ions. At the same time, tea polyphenols are plant extracts with high safety, which can reduce the toxicity of copper ions.

[0018] Amino acid compounds can interact with It forms a stable coordination complex, which prevents copper from precipitating and being adsorbed by impurities, making copper more stable, longer-lasting, and easier to penetrate biological membranes, thereby improving the stability and bioavailability of copper ions.

[0019] Hydroxycarboxylic acid salts can react with It forms a five-membered ring chelate, which prevents free copper ions from being adsorbed or precipitated by organic matter and sediment in the water, thereby improving the utilization rate of copper ions; it slowly releases active copper at bacterial target sites to achieve long-term antibacterial effect.

[0020] The method for preparing a copper-containing antibacterial solution provided by the present invention achieves a significant enhancement of the antibacterial effect by constructing a stable compound system with antibacterial enhancement components and copper-containing components. At the same time, it can reduce the amount of copper ions used, thereby improving safety while enhancing the antibacterial effect.

[0021] To balance antibacterial effect and safety, the present invention preferably uses a mass ratio of copper-containing component to antibacterial enhancement component of 1:(5-10). By controlling this ratio range, the copper-containing component and antibacterial enhancement component can fully exert their synergistic effect while ensuring antibacterial effect, and at the same time avoid the decrease in stability or increase in cost caused by imbalance of component ratio.

[0022] Furthermore, the preferred amino acid compound of the present invention is lysine.

[0023] Lysine is a basic amino acid with two amino groups and a carboxyl group, and it can react with... It forms a stable coordination complex, which prevents copper from precipitating and being adsorbed by impurities, making copper more stable, longer-lasting, and easier to penetrate biological membranes, thus improving the stability and bioavailability of copper ions. At the same time, amino acids can disrupt the metabolic processes of bacteria and synergistically inhibit bacterial growth with copper ions.

[0024] The preferred hydroxycarboxylic acid salt compound of this invention is potassium citrate.

[0025] The polycarboxyl groups of potassium citrate form a five-membered ring chelate with copper ions, preventing free copper ions from being adsorbed or precipitated by organic matter and sediment in the water, thus improving copper ion utilization. It also slowly releases active copper at bacterial target sites, achieving long-lasting antibacterial effects. Furthermore, the potassium citrate-copper complex is negatively charged, readily adsorbing onto the negatively charged outer membrane of bacteria, disrupting cell membrane integrity. Once inside the cell, copper ions bind to the sulfhydryl groups of enzymes, inhibiting the respiratory chain and energy synthesis, and disrupting DNA replication and protein synthesis. Finally, potassium citrate can chelate within biofilms... Stable cations disrupt the extracellular matrix, helping copper ions penetrate mature biofilms, kill deep-seated bacteria, and enhance antibacterial effects.

[0026] The antibacterial enhancing components in this invention can be used alone or in combination. Preferably, when tea polyphenols are used alone as the antibacterial enhancing component, the mass-volume concentration of tea polyphenols in the copper-containing antibacterial solution is controlled at 0.2%. Preferably, when lysine is used alone as the antibacterial enhancing component, the mass-volume concentration of lysine in the copper-containing antibacterial solution is controlled at 0.2%. Preferably, when potassium citrate is used alone as the antibacterial enhancing component, the mass-volume concentration of potassium citrate in the copper-containing antibacterial solution is controlled at 0.2%.

[0027] In this invention, when the amino acid compound lysine and the hydroxycarboxylic acid compound potassium citrate are combined, the mass-to-volume ratio of the two is 1:(1-5), at which point the synergistic antibacterial effect is optimal.

[0028] To ensure the antibacterial effect, the antibacterial enhancing components of this invention preferably include tea polyphenols, amino acid compounds, and hydroxycarboxylic acid salt compounds.

[0029] Specifically, when the antibacterial enhancing components include tea polyphenols, amino acid compounds, and hydroxycarboxylate compounds, mixing the copper-containing components with the antibacterial enhancing components includes: mixing the copper-containing components with the hydroxycarboxylate compounds, and then sequentially adding the amino acid compounds and tea polyphenols to obtain a copper-containing antibacterial solution.

[0030] Taking lysine as an amino acid compound and potassium citrate as a hydroxycarboxylic acid salt compound as an example, the preparation process involves first adding potassium citrate and... A stable five-membered ring chelate is formed to prevent copper adsorption, precipitation, and inactivation, and the pH is adjusted to maintain a mild and stable system. Lysine is then added, which forms a secondary coordination complex with copper to improve bioavailability. Finally, tea polyphenols are added, providing phenolic hydroxyl groups to interact with copper. Weak coordination reduces copper oxidation precipitation, and it also disrupts bacterial cell membranes and inhibits intracellular enzymes, thus preventing precipitation.

[0031] When the preferred antibacterial enhancing components of this invention include tea polyphenols, amino acid compounds, and hydroxycarboxylate compounds, the mass ratio of the hydroxycarboxylate compounds, amino acid compounds, and tea polyphenols is 1:1:1. Specifically, the mass-volume concentration of lysine in the copper-containing antibacterial solution is preferably controlled at 0.05%~1%, more preferably 0.05%~0.1%; the mass-volume concentration of potassium citrate in the copper-containing antibacterial solution is preferably controlled at 0.05%~1%, more preferably 0.05%~0.1%; and the mass-volume concentration of tea polyphenols in the copper-containing antibacterial solution is preferably controlled at 0.05%~1%, more preferably 0.05%~0.1%, to maximize the synergistic effect of the three antibacterial enhancing components and the copper-containing component.

[0032] This invention optimizes the performance of the antibacterial solution from multiple dimensions, such as improved stability, enhanced antibacterial efficacy, and improved environmental adaptability, through the synergistic effect of the three components in the antibacterial enhancement component, achieving a technical effect of 1+1+1>3. Furthermore, by adopting a stepwise mixing method, each component can sequentially complex or react with copper ions, avoiding the reaction disorder that may be caused by mixing multiple components at the same time, and further improving the stability and homogeneity of the solution.

[0033] The copper-containing component of this invention preferably includes copper gluconate; as an organic copper salt, copper gluconate has better water solubility and biocompatibility than inorganic copper salts, and can reduce irritation to the skin and mucous membranes while ensuring antibacterial effect.

[0034] Specifically, the copper gluconate concentration in the copper-containing antibacterial solution is preferably controlled at 0.005%~0.25% by mass and more preferably 0.025%~0.05%. This concentration range ensures the basic antibacterial effect while avoiding the toxicity of high concentrations of copper ions, and at the same time provides a basis for the synergistic effect of the antibacterial enhancement components.

[0035] To improve antibacterial performance and system stability, the copper-containing antibacterial solution provided by this invention may further include solvents, co-solvents, and other components. Specifically, taking antibacterial enhancing components including tea polyphenols, amino acid compounds, and hydroxycarboxylate compounds as an example, the preparation method of the copper-containing antibacterial solution in this invention can be carried out according to the following process: S1: According to the formula, copper gluconate, potassium chloride and propylene glycol are mixed to form mixture I. The whole system is carried out at 40±2℃. The purpose of adding potassium chloride in this step is to regulate osmotic pressure, and the purpose of adding propylene glycol is to retain moisture. S2: Add lauryl glucoside to mixture I and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add potassium citrate to mixture II and sonicate intermittently (30s / 10s) for 10 minutes to form mixture III; S4: Add lysine to mixture III and sonicate intermittently (30s / 10s) for 10 minutes to form mixture IV; S5: Add tea polyphenols to mixture IV and sonicate intermittently (30s / 10s) for 10 minutes to form mixture V; S6: Add Tween 80, a co-solvent, to mixture V to obtain a copper-containing antibacterial solution.

[0036] This invention achieves significantly enhanced antibacterial effects by constructing a stable compound system. In this system, the antibacterial enhancing components synergistically interact with copper ions, reducing copper ion dosage by 30%-60% while increasing the antibacterial rate by ≥20% compared to single copper-containing solutions. This solves the problems of insufficient antibacterial effect or excessive toxicity of single copper-containing solutions. The compound system is highly safe and widely applicable. The selected antibacterial enhancing components (especially natural plant extracts) are highly safe and can reduce the toxicity of copper ions, making it suitable for scenarios with high safety requirements, such as medical disinfection, food preservation, and antibacterial treatment of infant products. The compound system allows for adjustment of component ratios for different strains, exhibiting strong versatility. It also demonstrates good stability and repeatability. This invention clearly defines the types, ratios, and preparation methods of the compound components. The standardized experimental procedure ensures an experimental coefficient of variation ≤5%. The compound system can be stably stored at room temperature, facilitating industrial application.

[0037] Another object of the present invention is to provide a copper-containing antibacterial solution, which is prepared by the method described above for preparing a copper-containing antibacterial solution.

[0038] The copper-containing antibacterial solution provided by this invention achieves a significant enhancement of antibacterial effect by constructing a stable compound system with antibacterial enhancement components and copper-containing components. At the same time, it can reduce the amount of copper ions used, thereby improving safety while enhancing the antibacterial effect.

[0039] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, specific embodiments of the present invention will be described in detail below. Example 1

[0040] S1: Mix 0.25g copper gluconate, 8g potassium chloride and 8mL propylene glycol to form mixture I. The whole system is carried out at 40±2℃. S2: Add 8g of lauryl glucoside to mixture I, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add 2g of tea polyphenols to mixture II and sonicate intermittently (30s / 10s) for 10 minutes to form mixture III; S4: Add 1 mL of Tween 80 cosolvent to mixture III, and finally make up the difference with purified water to obtain 1 L of copper-containing antibacterial solution. Example 2

[0041] S1: Mix 0.25g copper gluconate, 8g potassium chloride and 8mL propylene glycol to form mixture I. The whole system is carried out at 40±2℃. S2: Add 8g of lauryl glucoside to mixture I, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add 2g of potassium citrate to mixture II and sonicate intermittently (30s / 10s) for 10 minutes to form mixture III; S4: Add 1 mL of Tween 80 cosolvent to mixture III, and finally make up the difference with purified water to obtain 1 L of copper-containing antibacterial solution. Example 3

[0042] S1: Mix 0.25g copper gluconate, 8g potassium chloride and 8mL propylene glycol to form mixture I. The whole system is carried out at 40±2℃. S2: Add 8g of lauryl glucoside to mixture I, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add 2g of lysine to mixture II and sonicate intermittently (30s / 10s) for 10 minutes to form mixture III; S4: Add 1 mL of Tween 80 cosolvent to mixture III to obtain a copper-containing antibacterial solution. Example 4

[0043] S1: Mix 0.25g copper gluconate, 8g potassium chloride and 8mL propylene glycol to form mixture I. The whole system is carried out at 40±2℃. S2: Add 8g of lauryl glucoside to mixture I, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add 1g of potassium citrate to mixture II and sonicate intermittently (30s / 10s) for 10 minutes to form mixture III; S4: Add 1g of tea polyphenols to mixture III and sonicate intermittently (30s / 10s) for 10 minutes to form mixture IV; S5: Add 1 mL of Tween 80 cosolvent to mixture IV, and finally make up the difference with purified water to obtain 1 L of copper-containing antibacterial solution. Example 5

[0044] S1: Mix 0.25g copper gluconate, 8g potassium chloride and 8mL propylene glycol to form mixture I. The whole system is carried out at 40±2℃. S2: Add 8g of lauryl glucoside to mixture I, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add 1g of lysine to mixture II and sonicate intermittently (30s / 10s) for 10 minutes to form mixture III; S4: Add 1g of tea polyphenols to mixture III, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture IV; S5: Add 1 mL of Tween 80 cosolvent to mixture IV, and finally make up the difference with purified water to obtain 1 L of copper-containing antibacterial solution. Example 6

[0045] S1: Mix 0.25g copper gluconate, 8g potassium chloride and 8mL propylene glycol to form mixture I. The whole system is carried out at 40±2℃. S2: Add 8g of lauryl glucoside to mixture I, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add 1g of potassium citrate to mixture II and sonicate intermittently (30s / 10s) for 10 minutes to form mixture III; S4: Add 1g of lysine to mixture III and sonicate intermittently (30s / 10s) for 10 minutes to form mixture IV; S5: Add 1 mL of Tween 80 cosolvent to mixture IV, and finally make up the difference with purified water to obtain 1 L of copper-containing antibacterial solution. Example 7

[0046] S1: Mix 0.25g copper gluconate, 8g potassium chloride and 8mL propylene glycol to form mixture I. The whole system is carried out at 40±2℃. S2: Add 8g of lauryl glucoside to mixture I, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add 0.7g potassium citrate to mixture II and sonicate intermittently (30s / 10s) for 10 minutes to form mixture III; S4: Add 0.7g of lysine to mixture III and sonicate intermittently (30s / 10s) for 10 minutes to form mixture IV; S5: Add 0.7g of tea polyphenols to mixture IV and sonicate intermittently (30s / 10s) for 10 minutes to form mixture V; S6: Add 1 mL of Tween 80 cosolvent to mixture V, and finally make up the difference with purified water to obtain 1 L of copper-containing antibacterial solution.

[0047] Comparative Example 1 S1: Mix 0.25g copper gluconate, 8g potassium chloride and 8mL propylene glycol to form mixture I. The whole system is carried out at 40±2℃. S2: Add 8g of lauryl glucoside to mixture I, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add 1 mL of Tween 80 cosolvent to mixture II, and finally make up the difference with purified water to obtain 1 L of copper-containing antibacterial solution.

[0048] Comparative Example 2 S1: Mix 0.25g copper gluconate, 8g potassium chloride and 8mL propylene glycol to form mixture I. The whole system is carried out at 40±2℃. S2: Add 8g of lauryl glucoside to mixture I, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture II; S3: Add 0.7g potassium citrate, 0.7g lysine, and 0.7g tea polyphenols to mixture II simultaneously, and sonicate intermittently (30s / 10s) for 10 minutes to form mixture III; S4: Add 1 mL of Tween 80 cosolvent to mixture III, and finally make up the difference with purified water to obtain 1 L of copper-containing antibacterial solution.

[0049] The antibacterial properties of the copper-containing antibacterial solutions prepared in each embodiment and comparative example were tested: (1) After the copper-containing antibacterial solutions prepared in each example and comparative example are prepared, they are sterile filtered through a 0.22 μm filter membrane to remove impurities and microbial contamination.

[0050] (2) Preparation of bacterial suspension: The target strain (Escherichia coli CMCC 11229 and Staphylococcus aureus CMCC 6538) was inoculated into tryptic soy broth (TSB) and activated to the logarithmic growth phase. The suspension was diluted with sterile physiological saline, and colonies were counted. Freshly cultured test bacteria were then used to prepare a suspension with a concentration of approximately [missing value] using 0.9% sterile sodium chloride solution. Bacterial suspension.

[0051] (3) Inoculation and reaction of test samples: Take sterile test tubes and add 5 mL of test sample to each tube. Prepare two parallel tubes for each test bacterium. Add 50 μL of the above working bacterial suspension to each tube of test sample to make the initial inoculation concentration approximately [missing information]. Vortex for 5-10 seconds to ensure thorough mixing, then immediately place the test tube in a 20-25°C incubator to begin the reaction. Samples are taken at 0 hours (immediately), 2 hours, 6 hours, and 24 hours after contact for viable cell counting.

[0052] (4) Viable cell count: Immediately after reaching each predetermined time point, remove the test tube and vortex mix. Take 0.5 mL of sample and add it to 4.5 mL of validated neutralizing agent: DEB, and perform a 10-fold serial dilution (dilution to...). , , , , , To stop the antibacterial reaction and neutralize residual antibacterial agents, select 3-5 suitable dilutions. Take 1.0 mL of each dilution into a sterile Petri dish, pour in approximately 15-20 mL of TSA tryptic soy peptone solid medium cooled to 45-50°C, mix well, and allow to solidify. Prepare two parallel Petri dishes for each dilution.

[0053] Invert the petri dish and incubate it in a 30-35°C incubator for 48-72 hours.

[0054] Control group setup: Positive control for test bacteria (initial concentration confirmation): Replace the test sample with diluent, and take samples at 0 hours to count and confirm the initial inoculum concentration. Negative control for product: Pour 1 mL of the mixture of test sample and neutralizing agent into TSA and check the product sterility. Negative control for diluent / neutralizing agent: Pour 1 mL of diluent / neutralizing agent into TSA and check the sterility. Negative control for culture medium: Pour into an uninoculated TSA plate and check the sterility of the culture medium.

[0055] The results of the antibacterial test are shown in Table 1.

[0056] Table 1 Based on the above-described preferred embodiments of the present invention, and through the foregoing description, those skilled in the art can make various changes and modifications without departing from the inventive concept. The technical scope of this invention is not limited to the contents of the specification, but must be determined according to the scope of the claims.

Claims

1. A method for preparing a copper-containing antibacterial solution, characterized in that, The method includes the following steps: mixing a copper-containing component with an antibacterial enhancing component to obtain a copper-containing antibacterial solution; wherein the antibacterial enhancing component is selected from at least one of tea polyphenols, amino acid compounds, and hydroxycarboxylic acid salt compounds.

2. The method for preparing the copper-containing antibacterial solution as described in claim 1, characterized in that, The mass ratio of the copper-containing component to the antibacterial enhancement component is 1:(5-10).

3. The method for preparing the copper-containing antibacterial solution as described in claim 1, characterized in that, The amino acid compounds include lysine.

4. The method for preparing the copper-containing antibacterial solution as described in claim 1, characterized in that, The hydroxycarboxylic acid salts include potassium citrate.

5. The method for preparing the copper-containing antibacterial solution according to any one of claims 1-4, characterized in that, The antibacterial enhancing components include tea polyphenols, amino acid compounds, and hydroxycarboxylic acid salt compounds.

6. The method for preparing the copper-containing antibacterial solution as described in claim 5, characterized in that, Mixing copper-containing components with antibacterial enhancing components includes: mixing copper-containing components with hydroxycarboxylic acid salt compounds, and then sequentially adding amino acid compounds and tea polyphenols to obtain the copper-containing antibacterial solution.

7. The method for preparing the copper-containing antibacterial solution as described in claim 6, characterized in that, The mass ratio of the hydroxycarboxylic acid salt compound, the amino acid compound, and the tea polyphenol is 1:1:

1.

8. The method for preparing the copper-containing antibacterial solution as described in claim 5, characterized in that, The copper-containing component includes copper gluconate.

9. A copper-containing antibacterial solution, characterized in that, The solution is prepared by the method described in any one of claims 1-8.

Images