CN114569630A

CN114569630A - Disinfecting and sterilizing liquid, its production method and use

Info

Publication number: CN114569630A
Application number: CN202011387786.9A
Authority: CN
Inventors: 黄柏川; 黄桂新; 黄庭奕
Original assignee: Quanpeng Technology Co ltd
Current assignee: Quanpeng Technology Co ltd
Priority date: 2020-12-02
Filing date: 2020-12-02
Publication date: 2022-06-03

Abstract

The embodiment of the invention provides a disinfection and sterilization liquid and a manufacturing method thereof. The disinfecting and sterilizing liquid is prepared by mixing sodium hypochlorite and weak acid. The effective chlorine concentration of the disinfection and sterilization liquid is more than 25ppm and less than or equal to 200ppm, and the effective chlorine of the disinfection and sterilization liquid is only from hypochlorous acid. The oxidation-reduction potential of the disinfecting and sterilizing liquid is 1100 to 1500 millivolts. The disinfection and sterilization liquid provided by the embodiment of the invention has better sterilization capability and sterilization speed, and has no toxicity and irritation to organisms.

Description

Disinfecting and sterilizing liquid, its production method and use

Technical Field

The present invention relates to a disinfectant and sterilizing liquor and its preparation method.

Background

A common method of manufacturing sterilizing solutions involves the electrolysis of sodium chloride solution or dilute hydrochloric acid. The electrolysis of sodium chloride solution can generate alkaline sodium hypochlorite, the electrolysis of dilute hydrochloric acid can generate acidic hypochlorous acid, and the sodium hypochlorite and the hypochlorous acid have the functions of disinfection and sterilization.

However, sodium hypochlorite is corrosive and irritant and is not suitable for use in living organisms or humans. Although the electrolysis of sodium chloride solution can produce high concentration sodium hypochlorite, sodium chloride remains therein and is undesirable for medical use. On the other hand, electrolytic dilute hydrochloric acid generates hypochlorous acid, chlorous acid, chloric acid, and the like. In the mixture generated by electrolyzing dilute hydrochloric acid, the oxidation capacity of hypochlorous acid is higher, the oxidation-reduction potential is higher, and the disinfection and sterilization effects are better. However, the concentration of hypochlorous acid generated by electrolyzing dilute hydrochloric acid is low, resulting in poor sterilization effect. When hypochlorous acid having a high concentration is produced, it is energy-consuming in production, and the product contains chlorous acid, chloric acid, etc. in addition to hypochlorous acid, and the ratio of the components is easily affected by the electrode and the electrolytic environment, and is difficult to control. Therefore, it is one of the current research directions to produce a sterilizing solution suitable for living organisms and having an excellent sterilizing ability while considering both the manufacturing cost and the product quality.

Disclosure of Invention

The disinfection and sterilization liquid and the manufacturing method thereof provided by the embodiment of the invention solve the problem that the sterilization liquid containing hypochlorous acid cannot further improve the sterilization capability and the sterilization speed.

One embodiment of the present invention provides a disinfectant and sterilization solution comprising sodium ions, a first weak acid, a second weak acid, and water. The first weak acid and the second weak acid are dissimilar weak acids, and the second weak acid is hypochlorous acid. The effective chlorine concentration of the disinfection and sterilization liquid is more than 25ppm and less than or equal to 200ppm, and the effective chlorine of the disinfection and sterilization liquid is only from hypochlorous acid. The oxidation-reduction potential of the disinfecting and sterilizing liquid is 1100 to 1500 millivolts.

One embodiment of the present invention provides a disinfecting and sterilizing solution that substantially comprises sodium ions, a first weak acid, a second weak acid, and water. The first weak acid and the second weak acid are dissimilar weak acids, and the second weak acid is hypochlorous acid. The effective chlorine concentration of the disinfection and sterilization liquid is more than 25ppm and less than or equal to 200 ppm. The oxidation-reduction potential of the disinfecting and sterilizing liquid is 1100 to 1500 millivolts.

One embodiment of the present invention provides a method for producing a disinfecting and sterilizing solution, comprising mixing sodium hypochlorite, a first weak acid and water to react to form the disinfecting and sterilizing solution. The disinfection sterilization liquid comprises sodium ions, a first weak acid, a second weak acid and water. The first weak acid and the second weak acid are dissimilar weak acids, and the second weak acid is hypochlorous acid. The effective chlorine concentration of the disinfection and sterilization liquid is more than 25ppm and less than or equal to 200ppm, and the effective chlorine of the disinfection and sterilization liquid is only from hypochlorous acid. The oxidation-reduction potential of the disinfecting and sterilizing liquid is 1100 to 1500 millivolts.

The invention also provides application of the disinfection and sterilization liquid in preparing a composition for treating wounds, wherein the disinfection and sterilization liquid is the disinfection and sterilization liquid.

The invention also provides a use of the disinfection and sterilization liquid for preparing a composition for treating infection, wherein the disinfection and sterilization liquid is the disinfection and sterilization liquid.

According to the disinfection solution and the manufacturing method thereof provided by one embodiment of the invention, by mixing sodium hypochlorite and weak acid, the disinfection solution containing high-purity hypochlorous acid can be provided, and the disinfection solution does not contain chloride ions, chloric acid or chlorous acid. That is, the effective chlorine of the disinfecting and sterilizing liquid according to one embodiment of the present invention comes only from hypochlorous acid having a high oxidizing ability. Compared with hypochlorous acid sterilizing liquid prepared by electrolyzing dilute hydrochloric acid, the sterilizing liquid provided by the invention has higher oxidation-reduction potential under the same effective chlorine concentration. Therefore, the disinfection and sterilization liquid according to one embodiment of the invention has better sterilization capability and sterilization speed, and has no toxicity and irritation to organisms. Furthermore, the disinfectant sterilizing liquid of one embodiment of the invention can be used for preparing a composition for treating wounds or treating infection. Treating the wound may include disinfecting the wound resulting from the surgical procedure, the decubitus ulcer, or the burn and scald. Treating the infection may comprise disinfecting infection caused by surgery, peritonitis, bedsores, or burns, and may comprise disinfecting infection caused by bacteria or viruses. By performing wound treatment or infection treatment using the disinfectant sterilizing liquid according to one embodiment of the present invention, infection can be prevented or inhibited, thereby accelerating wound healing.

Drawings

FIG. 1 shows the cell morphology under microscope after 48 hours of treatment with disinfectant and bactericidal solution with different effective chlorine concentrations.

FIG. 2 shows the cell morphology under microscope of virus-infected cells after treatment with and without a disinfectant and disinfectant solution.

FIG. 3 shows the cell growth under microscope of the blank control group, the positive control group, the negative control group and the test group in the in vitro cytotoxicity test.

Fig. 4A to 4C are photographs of the backs of rabbits 1 hour, 24 hours, and 48 hours after removal of gauze in the skin irritation test, respectively.

FIG. 5 shows the change of the effective chlorine concentration of the fifth and sixth groups with time in the stability test.

FIG. 6 shows the change of the effective chlorine concentration with time in the test group and the control group in the stability test.

Detailed Description

The detailed features and advantages of the invention are described in the following detailed description, which is sufficient for anyone skilled in the art to understand the technical contents of the invention and to implement the invention, and the objects and advantages related to the invention can be easily understood by anyone skilled in the art based on the disclosure of the present specification, the claims and the accompanying drawings. The following examples are intended to illustrate aspects of the present invention in further detail, but are not intended to limit the scope of the invention in any way.

First, a disinfecting and sterilizing liquid according to an embodiment of the present invention will be described. In one embodiment, the sanitizer includes sodium ions, a first weak acid, a second weak acid, and a solvent. In other words, the disinfecting and sterilizing solution according to an embodiment of the present invention basically includes sodium ions, a first weak acid, a second weak acid and a solvent. The effective chlorine concentration of the disinfection and sterilization liquid is more than 25ppm and less than or equal to 200ppm, and the effective chlorine of the disinfection and sterilization liquid is only from hypochlorous acid. The oxidation-reduction potential of the disinfecting and sterilizing liquid is 1100 to 1500 millivolts. The first weak acid and the second weak acid are dissimilar weak acids and the second weak acid is hypochlorous acid. In other embodiments, the first weak acid may have a pKa of 4 to 7, and the weak acid having a pKa of 4 to 7 may be, for example, citric acid, carbonic acid, or acetic acid. The solvent is water. In other embodiments, the pH of the disinfecting solution may be 4 to 6. In other embodiments, the disinfecting sterilizing solution may not contain chloride ions, chlorous acid, or chloric acid. In other embodiments, the available chlorine concentration of the disinfecting and sterilizing solution can be greater than 100ppm and less than or equal to 200ppm, and the oxidation-reduction potential of the disinfecting and sterilizing solution can be 1300 to 1500 millivolts.

Next, a method for producing a disinfecting and sterilizing liquid according to an embodiment of the present invention will be described. In one embodiment, a method of making a disinfecting solution includes mixing sodium hypochlorite, a first weak acid, and a solvent to react to form the disinfecting solution as described above. Specifically, sodium hypochlorite is added into pure water to form a sodium hypochlorite aqueous solution, then the dosage of the selected first weak acid is calculated according to the pKa, the target pH value and the target effective chlorine concentration, and the first weak acid is added into the sodium hypochlorite aqueous solution to perform gradual reaction to form a homogeneous solution. Measuring the pH value and the effective chlorine concentration of the generated solution, comparing the pH value and the effective chlorine concentration with the target pH value and the target effective chlorine concentration, calculating the compensation dosage of the sodium hypochlorite, and adding the sodium hypochlorite into the solution to adjust the pH value and the effective chlorine concentration. Measuring the redox potential of the resulting solution, comparing with a target redox potential, calculating a selected adapted dose of a first weak acid, and adding the first weak acid to adjust the redox potential. The pH, available chlorine concentration and oxidation-reduction potential are repeatedly adjusted until target values are met. By mixing the first weak acid with sodium hypochlorite, not only hypochlorous acid but also conjugate base of the first weak acid can be generated. The first weak acid and the conjugate base thereof can form a buffer solution with a buffering effect, and can stabilize the pH value of the disinfection and sterilization liquid, thereby maintaining the stability of the disinfection and sterilization liquid. Therefore, the method for manufacturing the sterilizing liquid according to one embodiment of the present invention can eliminate the need for an additional buffer. The first weak acid may have a pKa of 4 to 7, and the weak acid having a pKa of 4 to 7 is, for example, citric acid, carbonic acid, or acetic acid. The solvent is water. The weight ratio of sodium hypochlorite to the first weak acid may be 1.0:0.5 to 1.0: 1.5. In other embodiments, the first weak acid may be carbonic acid, and the weight ratio of sodium hypochlorite to carbonic acid may be 1: 0.87.

Table 1 shows the composition ratio, effective chlorine concentration, oxidation-reduction potential and pH of each example. The disinfectant and sterilizing solutions of examples 1 to 3 were 5 liters solutions prepared from 10% by weight aqueous sodium hypochlorite solution, 1% by weight aqueous carbonic acid solution and water, wherein pKa of carbonic acid was 6.35. The disinfecting and sterilizing solutions of examples 1 to 3 do not contain chloride ions, chlorous acid and chloric acid. The available chlorine concentration (unit: ppm) was measured by using a available chlorine detector (SIBAAS, ClO) with reference to an operation manual attached to the manufacturer. The operation steps are briefly described as follows: taking colorless and transparent water to be measured, setting a zero point by using an effective chlorine detector, adding a potassium iodide reagent, measuring the concentration of the effective chlorine, keeping the solution at a stable level in the measuring process, and measuring within a specified time. The oxidation-reduction potential (unit: millivolts (mV)) was measured using an ORP detector (ORP TESTER, ORP-986) with reference to the manufacturer's operating manual. The operation steps are briefly described as follows: and taking colorless and transparent water to be detected, putting the ORP detector into the water to be detected, standing for 3-5 minutes until the numerical value is stable, setting a reference value, putting the ORP detector into the solution to be detected, standing for 3-5 minutes until the numerical value is stable, and recording the measured value.

TABLE 1

As can be seen from table 1, examples 1 to 3 have high oxidation-reduction potentials, indicating good bactericidal ability and bactericidal rate.

Examples 1 to 3 are prepared by mixing sodium hypochlorite and carbonic acid at a weight ratio of about 1:0.87, but not limited thereto, and the composition and ratio of the weak acid can be determined according to the desired pH, oxidation-reduction potential, and effective chlorine concentration. Table 2 shows the oxidation-reduction potentials at different pH values of the disinfecting and sterilizing solution of the example of the present invention at the same effective chlorine concentration and the pH values and oxidation-reduction potentials of the comparative examples. In Table 2, the effective chlorine concentration of example 4 is 33 ppm; the available chlorine concentration of example 5 was 48 ppm; the effective chlorine concentration of examples 6 to 8 was 107 ppm; the effective chlorine concentrations of examples 9 to 11 were 147 ppm; the effective chlorine concentration of examples 12 to 14 was 188 ppm. Comparative example 1 is a commercially available hypochlorous acid antibacterial solution of brand a, which was prepared by an electrolytic method and had an effective chlorine concentration of 20 ppm; comparative example 2 is a commercially available hypochlorous acid antibacterial liquid of brand B, which was prepared by an electrolytic method and had an effective chlorine concentration of 44 ppm; comparative example 3 is a commercially available hypochlorous acid antibacterial solution of brand C, which was prepared by an electrolytic method and had an effective chlorine concentration of 20 ppm.

TABLE 2

As is clear from Table 2, increasing the effective chlorine concentration increases the oxidation-reduction potential. At the same effective chlorine concentration, lowering the pH increases the oxidation-reduction potential. In addition, in examples 4 and 5, compared with the commercially available hypochlorous acid sterilizing liquid, it is understood that the oxidation-reduction potentials of the examples are all larger than that of the comparative example. Therefore, compared with the commercially available sterilization liquid prepared by electrolysis, the sterilization liquid of the present embodiment prepared by mixing sodium hypochlorite and weak acid has higher oxidation-reduction potential, and thus has better sterilization capability and sterilization speed, because the effective chlorine of the sterilization liquid of the present embodiment is only from hypochlorous acid with higher oxidation-reduction potential.

The following description will be made of the disinfection and sterilization liquid according to an embodiment of the present invention, in order to confirm that the disinfection and sterilization liquid according to an embodiment of the present invention has excellent antibacterial and antiviral abilities, is harmless and nontoxic to living bodies, and has good stability, by performing an antibacterial test, an antiviral test, an in vitro cytotoxicity test, a skin irritation test, an eye irritation test, an inhalation toxicity test, and a stability test.

[ antibacterial test ]

The results of the antibacterial test using the disinfecting and sterilizing solution of different effective chlorine concentrations are described below. The test was entrusted to SGS Taiwan inspection science and technology, Inc. The effective chlorine concentration of the disinfection and sterilization liquid is 30ppm, 50ppm and 100 ppm. The test results are shown in table 3.

TABLE 3

As is clear from Table 3, the antibacterial activity of the disinfecting and sterilizing solution having the effective chlorine concentrations of 30ppm, 50ppm and 100ppm was 99.9% or more, indicating that the disinfecting and sterilizing solution indeed has excellent antibacterial activity. In addition, since the disinfecting and sterilizing solution can effectively resist bacteria even when the available chlorine concentration is 30ppm, 50ppm or 100ppm, it is presumed that the disinfecting and sterilizing solution can have excellent antibacterial ability even when the available chlorine concentration is 100ppm or more.

[ antiviral test ]

The results of the antiviral test using the disinfecting and sterilizing solution are described below. This test was conducted by the science and technology company Limited, the United states of the Committee for testing. This test is described in ASTM E1053-97: 2002 and ISO 10993-5: 2009 disinfectant and bactericidal solutions were evaluated for their antiviral efficacy against enterovirus EV71 type. In order to avoid the problem that the cytopathic effect is caused by a virus or a test substance, the cytotoxicity of the test substance is evaluated before the antiviral test, and the antiviral test is performed after the concentration of the test substance that is not toxic to the cells is found. The test is divided into a cytotoxicity evaluation test and an antiviral test.

The cytotoxicity assessment assay was performed as MTT assay. In detail, the toxicity of the disinfectant and sterilizing solution to the cells was evaluated by comparing the difference in the number of the living cells between the test group and the control group by measuring the absorbance of MTT reduced by the living cells. The test group is cells treated by a disinfecting and sterilizing solution, and the disinfecting and sterilizing solution comprises 1/10-1/640 diluted by a cell culture solution with the effective chlorine concentration of 200ppm so that the effective chlorine concentration is 0.625 ppm-20 ppm. The control group was cells treated with cell culture broth. The test cell line was human embryonic kidney epithelial tumor cell (human embryonic kidney epithelial tumor cell line, RD cell, ATCC number: CCL-136). The test results are shown in table 4 and fig. 1. FIG. 1 shows the cell morphology under microscope after 48 hours of treatment with disinfectant and bactericidal solution with different effective chlorine concentrations. In fig. 1, a: control group, B: 20ppm, C: 10ppm, D: 5ppm, E: 2.5ppm, F: 1.25ppm, G: 0.625ppm, H: 0.3125 ppm. In table 4, cell viability less than 70% was judged to be cytotoxic, and a p-value less than 0.05 indicated a significant difference compared to the control group.

TABLE 4

As can be seen from Table 4, the cell viability of the test groups was all greater than 70%, indicating that the disinfectant and bactericidal solution with effective chlorine concentration of 0.625ppm to 20ppm was not toxic to cells, and the p-values were all greater than 0.05, indicating that the effective chlorine concentration did not significantly affect the cell viability. Furthermore, as can be seen from FIG. 1, the cells (B) to (H) in the test group were not significantly different in cell morphology and growth density from the control group cell (A). Therefore, the disinfection and sterilization solution with the effective chlorine concentration of 0.625ppm to 20ppm has no influence on cell growth and does not generate cytotoxicity.

The antiviral test comprises treating virus with or without disinfectant, diluting with cell culture medium, infecting cells, comparing pathological changes of cells, and testing according to Reed-calculation of TCID by Muench method₅₀The virus titer. TCID per ml₅₀Viral Titer (TCID)₅₀mL) represents the ability of the virus solution per mL to infect cells. The drop in the log of viral titer represents the ability of the test substance to destroy the virus. Log reduction in viral titer₁₀TCID of control group₅₀/mL)-(log₁₀TCID of test group₅₀/mL). The disinfectant has a virus killing capacity of 4log or more according to the US EPA regulations₁₀. In this test, the strain was Enterovirus EV71 type (Enterovirus-71). The results of the experiment are shown in table 5 and fig. 2. In table 5, the dilution factor is the concentration factor of the serial dilutions of the virus liquid; the control group is cells infected by virus without being treated by the disinfection and sterilization solution; the test group is cells infected by virus treated by 20ppm of disinfection and sterilization solution; the control group is cells without virus infection; "+" indicates the number of cytopathies present; "-" no cytopathic effects occurred. FIG. 2 shows the cell morphology under microscope of virus-infected cells after treatment with and without a disinfectant and disinfectant solution. In fig. 2, a: control group, B: control 10-fold dilution, C: control group 10²Dilution by time, D: control group 10³Fold dilution, E: control group 10⁴Double dilution, F: control group 10⁵Dilution by time, G: control group 10⁶Dilution by time, H: test group 10-fold dilution, I: test group 10²Fold dilution, J: test group 10³Dilution by time, K: test group 10⁴And (5) diluting by times.

TABLE 5

As can be seen from table 5 and fig. 2, the cell morphology of the control group (a) was normal, the control groups (B) to (E) showed significant cytopathic effect at higher virus concentration, and the cell morphology and growth density of the test groups (H) to (K) were not significantly different from those of the control group (a). The test group did not find the cytopathic condition even at the lowest dilution factor, and the virus was considered to have lost activity, indicating that the disinfectant and bactericidal solution can indeed eliminate the virus activity and reduce the diseaseThe ability of the virus to infect cells. Calculation of TCID by Reed-Muench method₅₀The virus titer of the control group was 4.47X 10 per ml⁵The virus titer per ml of the test group was less than 10^0.5The reduction value of the logarithm of the virus titer is more than 5 logs₁₀The disinfectant meeting the United states EPA regulations should have a virus killing capacity of greater than or equal to 4log₁₀The requirements of (1). Therefore, the disinfection and sterilization liquid of the embodiment of the invention has the capability of killing viruses. In addition, since the disinfecting and sterilizing solution can effectively resist viruses when the effective chlorine concentration is 20ppm, it is presumed that the disinfecting and sterilizing solution has good antiviral ability when the effective chlorine concentration is more than 20 ppm.

[ antibacterial and antiviral test of a disinfecting/sterilizing solution having an effective chlorine concentration of 30ppm ]

The following are the results of the tests of the disinfection solution of the present invention with 30ppm of available chlorine for antibacterial and antiviral purposes. The test was entrusted to SGS taiwan inspection science and technology limited (hereinafter abbreviated as SGS), taimei inspection science and technology limited (hereinafter abbreviated as taimei technology), and the food industry development research institute of the financial group act (hereinafter abbreviated as food research institute). The test results are shown in table 6.

TABLE 6

As shown in table 6, the inhibition rate and inhibition rate of the disinfectant and bactericidal solution with 30ppm of available chlorine concentration are over 99% as confirmed by various public trust organizations, which indicates that the disinfectant and bactericidal solution has excellent inhibition capability against drug-resistant bacteria (such as drug-resistant staphylococcus aureus, acinetobacter baumannii), spore bacteria (such as bacillus subtilis, clostridium), bacteria and fungi (such as staphylococcus aureus, escherichia coli, pseudomonas aeruginosa, candida albicans, salmonella enterica, streptococcus mutans, klebsiella pneumoniae, propionibacterium acnes, aspergillus niger, listeria) and viruses such as enterovirus EV-71. In addition, since the disinfectant and bactericidal solution can effectively resist bacteria and viruses when the effective chlorine concentration is 30ppm, it is presumed that the disinfectant and bactericidal solution can have excellent antibacterial ability even when the effective chlorine concentration is more than 30 ppm.

[ antiviral test of sterilizing solution having an effective chlorine concentration of 100ppm ]

The following is the test result of the anti-virus of the disinfectant and bactericidal liquid with the effective chlorine concentration of 100ppm of the embodiment of the invention. This test was entrusted to Microbac Laboratories and was conducted in accordance with ASTM E1053-20. The test viruses were Coronavirus (Severe acid Respiratory Syndrome Coronavir 2, SARA-CoV-2, COVID 19Virus) and Influenza Virus (Influenza A Virus, H1N 1). The test results are shown in table 7.

TABLE 7

As shown in table 7, the Log removal rate (Log10 Reduction) of 100ppm of available chlorine for the disinfectant and bactericidal solution was 1 and represented the inhibition rate (Percent Reduction) of 90%, both of which were confirmed by the international organization with public confidence, indicating that the disinfectant and bactericidal solution indeed has excellent inhibitory activity against both of the coronavirus and the influenza virus. In addition, since the disinfecting and sterilizing solution can effectively inhibit coronavirus or influenza virus even at an effective chlorine concentration of 100ppm, it is presumed that the disinfecting and sterilizing solution can have an excellent virus-inhibiting ability even at an effective chlorine concentration of 100ppm or more.

[ in vitro cytotoxicity assay ]

The results of in vitro cytotoxicity tests using the disinfecting and sterilizing solution having an effective chlorine concentration of 200ppm are described below. The test was entrusted to SGS Taiwan inspection science and technology Co., Ltd. According to the test, according to the specifications of ISO10993-5, ISO10993-12 and ASTM F895-11, the cell appearance and the monolayer cell growth density are qualitatively observed by an Agar dispersion method, and finally, the cell appearance and the monolayer cell growth density are scored to judge whether cytotoxicity is generated or not. The test body was mouse lung fibroblast cell line (mouse lung fibroblast cells, L929 cells, Food Industry Research and Development Institute, Strain No. BCRC 60091).

In detail, in this test, cells were classified into a blank control group, a positive control group, a negative control group, and a test group. The blank control group was treated with cell culture fluid. The positive control group was treated with a cell culture medium containing 2% DMSO. The negative control group was treated with phosphate buffer. The test groups were treated with 200ppm of a disinfectant and disinfectant solution. The test group and the control group are placed on the solidified medium and then placed in an incubator at 37 +/-1 ℃ for 24 +/-2 hours. After 24. + -.2 hours the test substance is position-marked and removed. Then 2 ml of neutral red solution was added for staining for 1 hour. After staining was complete, the neutral red solution was aspirated, and the cell appearance and monolayer cell density were observed and recorded under a microscope. Observing whether the growth condition of the cells in the area is abnormal, degenerated, dead or cytolytic. The test results are shown in table 8 and fig. 3. FIG. 3 shows the cell growth under microscope of the blank control group, the positive control group, the negative control group and the test group in the in vitro cytotoxicity test.

TABLE 8

	Treatment agent	Toxic reaction
			Blank control group	Cell culture solution	Is free of
Positive control group	Cell culture broth containing 2% DMSO	Of moderate degree
			Negative control group	Phosphate buffer	Is composed of
Test group	200ppm disinfecting and sterilizing liquid	Is free of

As can be seen from table 8 and fig. 3, the positive control group showed significant changes in cell density compared to the blank control group, and was judged to be toxic to cells. The negative control group and the test group showed no significant difference in cell density compared to the blank control group, indicating that these two groups were not toxic to cells. Therefore, the disinfection and sterilization liquid of the embodiment of the invention does not generate cytotoxicity to cells. In addition, since the disinfecting/sterilizing solution does not cause toxicity to cells even when the effective chlorine concentration is 200ppm or less, it is presumed that the disinfecting/sterilizing solution does not cause toxicity to cells even when the effective chlorine concentration is 200ppm or less.

[ skin irritation test ]

The results of the skin irritation test using the disinfectant and bactericidal solution having an effective chlorine concentration of 200ppm are described below. The test was entrusted to SGS Taiwan inspection science and technology Co., Ltd. The test species was New Zealand white rabbits. This test investigated whether the disinfectant and bactericidal solution of the example produced a local irritation reaction on the damaged skin of rabbits.

In detail, the rabbit was shaved locally on the back, the shaved area was sterilized with 75% alcohol, and then a broken cross-cut wound was drawn with a sterile needle. 0.5 ml of sterile solution was applied to the damaged area on the left side of the back of the rabbit with 2.5X 2.5 cm square sterile gauze as a test group, and the right side was used as a control group. After 4 hours, the plate was washed with distilled water. The test results are shown in fig. 4A to 4C. Fig. 4A to 4C are photographs of the backs of rabbits 1 hour, 24 hours, and 48 hours after removal of gauze in the skin irritation test, respectively.

As can be seen from fig. 4A to 4C, the skin reaction was observed after 1, 24 and 48 hours after the test, and the results showed that the test group area and the control group area had no clinical symptoms such as erythema, swelling, inflammation, erosion, other toxic reactions and the like, and no death of any animal. Therefore, the disinfectant and bactericidal liquid provided by the embodiment of the invention does not cause irritation reaction on the damaged skin of the rabbit. In addition, since the disinfecting and sterilizing solution does not cause a skin irritant reaction when the available chlorine concentration is 200ppm, it is presumed that the disinfecting and sterilizing solution does not cause a skin irritant reaction when the available chlorine concentration is 200ppm or less.

[ eye irritation test ]

The results of the eye irritation test using the disinfecting and sterilizing liquid having an effective chlorine concentration of 210ppm are described below. The test is carried out by a verification laboratory of the plastic industry technology center of the financial group legal persons. The test is based on ISO 10993-10 and OECD test guide 405 standard, and the disinfectant sterilizing liquid with the effective chlorine concentration of 210ppm is directly instilled to rabbit eyes to judge whether to cause irritation reaction. The test species was New Zealand white rabbits.

Specifically, 0.1 ml of the test substance and the control substance were applied to the left and right eyes in a single eye drop, and the eye reactions of the test group and the control group were observed after 1, 24, 48, and 72 hours. The test substance is a disinfectant and bactericidal solution with the effective chlorine concentration of 210ppm, and the control substance is physiological saline. After the addition, both eyes of each animal were examined at 1. + -. 0.1, 24. + -. 2, 48. + -. 2, 72. + -.2 hours, and judged for eye reaction. The test results are shown in table 9.

TABLE 9

As shown in Table 9, no eye lesions and symptoms were observed in the test group and the control group, and there was no difference between them, and no death of the rabbits occurred. Therefore, 0.1 ml of the disinfectant and bactericidal solution with the effective chlorine concentration of 210ppm is directly instilled to the eyes of the rabbits and does not generate irritation reaction. In addition, since the disinfecting solution does not cause eye irritation when the available chlorine concentration is 210ppm, it is presumed that eye irritation does not occur when the available chlorine concentration is 210ppm or less.

[ inhalation toxicity test ]

The results of the inhalation toxicity test using the disinfecting and sterilizing liquid having an effective chlorine concentration of 210ppm are described below. This test was conducted by Retard Biotechnology, Inc. Animal animals were 6 week old Sprague-Dawley (SD) rats. The special antibacterial atomizer is manufactured by Roc science and technology, Inc., and has the following batch number: WT-J9.

In detail, 10 male mice and 10 female mice were divided into a test group and a control group. The test substance was administered by repeated dose nebulization using an antibacterial special nebulizer for 14 days for 2 hours per day, and whether the test substance caused an acute inhalation response in rats was evaluated. After 14 days, all animals were sacrificed for dissection and observation for the associated lesions. In this test, a disinfectant and bactericidal solution having an effective chlorine concentration of 210ppm was administered to the test group, and RO water was administered to the control group. The test results are shown in table 10.

Watch 10

As can be seen from Table 10, no clinical symptoms and no death of rats were observed in both the test group and the control group during the test period. After the test, the rats were sacrificed and dissected, and no clinical lesions were observable in both the test and control groups. Furthermore, there was no significant difference in animal body weight and no significant difference in feed intake during the test period. Therefore, the disinfectant and bactericidal solution with the effective chlorine concentration of 210ppm does not cause acute inhalation toxicity reaction or death situation to the rats. In addition, since the disinfectant/bactericidal solution does not cause inhalation toxicity to rats when the available chlorine concentration is 210ppm or less, it is presumed that the disinfectant/bactericidal solution does not cause inhalation toxicity to rats when the available chlorine concentration is 210ppm or less.

[ stability test ]

First, the results of the stability test using the disinfecting and sterilizing liquid having an effective chlorine concentration of 85ppm will be described below. The test was performed by the Biochemical laboratory of the Panc science and technology corporation. In this test, 500 ml of the sterilizing solution sealed in a container made of LDPE was left for different periods of time, and then the concentration of available chlorine was measured, and the decomposition of each group of available chlorine concentrations at each time was observed. The available chlorine concentration (unit: ppm) was measured by using a available chlorine detector (SIBAAS, ClO) with reference to an operation manual attached to the manufacturer. The operation steps are briefly described as follows: taking colorless and transparent water to be measured, setting a zero point by using an effective chlorine detector, adding a potassium iodide reagent, measuring the concentration of the effective chlorine, keeping the solution at a stable level in the measuring process, and measuring within a specified time.

Specifically, the test was divided into six groups, the first group was left in a sealed state for 24 months, the second group was left in a sealed state for 18 months, the third group was left in a sealed state for 12 months, the fourth group was left in a sealed state for 6 months, the fifth group was measured weekly and sealed after the measurement, and the sixth group was measured daily and sealed after the measurement. The test results are shown in table 11 and fig. 5. FIG. 5 shows the change of the effective chlorine concentration of the fifth and sixth groups with time in the stability test.

TABLE 11

As is clear from Table 11, the disinfectant/bactericidal solution having an effective chlorine concentration of 85ppm maintained an effective chlorine concentration of 30ppm or more even after being left in a sealed state for 24 months. Under the condition of proper storage, the effective chlorine concentration of more than 40ppm can be maintained after 6 months. In the state of not being properly preserved, the effective chlorine concentration of the sterilizing liquid is decomposed to 0ppm after 57 days. As can be seen from FIG. 5, the available chlorine concentration of the disinfecting and sterilizing solution decreases with time, and the decomposition rate in the properly stored state can be much slower than that in the non-properly stored state. Because the effective chlorine concentration is more than 20ppm, the antiviral effect can be achieved, and the effective chlorine concentration is more than 30ppm, the disinfection and sterilization liquid with the effective chlorine concentration of 85ppm can still maintain the effective chlorine concentration of more than 30ppm after being placed for 24 months, which shows that the disinfection and sterilization liquid has good stability and still has excellent disinfection and sterilization effects after being placed for a long time under a sealed condition. In addition, since the disinfection solution having an effective chlorine concentration of 85ppm can maintain an effective chlorine concentration of 30ppm or more even after being left for 24 months, it is presumed that the disinfection solution having an effective chlorine concentration of 85ppm or more can be left for a longer period of time and can maintain an excellent disinfection effect.

Next, the results of comparison of the stability of the disinfecting/sterilizing liquid having an effective chlorine concentration of 53ppm with that of a commercially available sterilizing liquid will be described below. The experimental conditions were as described above except that the accelerated aging test was performed in a temperature-controlled manner in this test. In detail, each test sample was stored in the same container of 100 ml, and the lapse of time was simulated by leaving it at 55 ℃ and converted to equivalent days. Each test sample was measured for available chlorine concentration weekly and sealed after measurement.

In this test, the test group was a disinfectant sterilizing liquid of one embodiment of the present invention having an effective chlorine concentration of 53 ppm; control group 1 was a commercially available hypochlorous acid antibacterial solution of brand a, which was prepared by electrolysis and had an effective chlorine concentration of 20 ppm; control group 2 was a commercial B-brand hypochlorous acid antibacterial solution prepared by electrolysis and having an effective chlorine concentration of 44 ppm; control 3 was a commercial C-brand hypochlorous acid antibacterial solution prepared by electrolysis and had an effective chlorine concentration of 20 ppm. The test results are shown in table 12 and fig. 6. FIG. 6 shows the change of the effective chlorine concentration with time in the test group and the control group in the stability test.

TABLE 12

As is clear from Table 12 and FIG. 6, the decrease rate of the effective chlorine concentration was the slowest in the test group. It is understood that, in the state of being stored properly, the decomposition rate of hypochlorous acid in the test group is lower than that in the control group on the market, and it means that the disinfecting and sterilizing liquid according to the embodiment of the present invention has better stability and can be stored for a longer period of time than the disinfecting liquid sold on the market.

[ composition analysis of Disinfection/Sterilization solution having an effective chlorine concentration of 30ppm ]

The following is a composition analysis of the disinfecting and sterilizing liquid of the embodiment of the present invention having an effective chlorine concentration of 30 ppm. The test entrusts the detection of the SGS taiwan inspection science and technology corporation (hereinafter abbreviated as SGS) and the plastic industry technology development center of the fiscal treasury corporate (hereinafter abbreviated as plastic center). The detection items comprise plasticizers, eight major harmful heavy metals and nonmetal elements, chlorine or benzene containing toxic compounds and bacteria content, wherein the plasticizers comprise butyl phthalate benzyl ester (BBP), dibutyl phthalate (DBP), di (2-ethylhexyl) phthalate (DEHP), di-n-octyl phthalate (DNOP), di-n-pentyl phthalate (DnPP), di (2-methoxyethyl) phthalate (DMEP) and diisoamyl phthalate (DIPP); the eight harmful heavy metals and nonmetal elements comprise arsenic, lead, mercury, cadmium, selenium, chromium, antimony and barium; toxic compounds containing chlorine or benzene include toluene, xylene, benzene, ethylbenzene, styrene, methylene chloride, 1, 1-dichloroethane, 1, 2-dichloroethane, chloroform, 1,1, 1-trichloroethane, carbon tetrachloride, trichloroethylene, and tetrachloroethylene. The analysis results are shown in Table 13.

Watch 13

As can be seen from table 13, the disinfection and sterilization liquid having 30ppm of available chlorine concentration was confirmed by various public trust organizations to be free from plasticizer, eight major harmful heavy metals and non-metallic elements, toxic compounds containing chlorine or benzene, and bacteria, which indicates that the disinfection and sterilization liquid is indeed harmless and nontoxic to living bodies.

The antibacterial test, the antiviral test, the in vitro cytotoxicity test, the skin irritation test, the eye irritation test, the inhalation toxicity test and the stability test prove that the disinfection and sterilization liquid disclosed by the embodiment of the invention has excellent antibacterial and antiviral capabilities, is harmless and nontoxic to organisms and has good stability.

According to the disinfection solution and the manufacturing method thereof provided by one embodiment of the invention, by mixing sodium hypochlorite and weak acid, the disinfection solution containing high-purity hypochlorous acid can be provided, and the disinfection solution does not contain chloride ions, chloric acid or chlorous acid. That is, the effective chlorine of the disinfecting and sterilizing liquid according to one embodiment of the present invention comes only from hypochlorous acid having a high oxidizing ability. Compared with hypochlorous acid sterilizing liquid prepared by electrolyzing dilute hydrochloric acid, the sterilizing liquid provided by the invention has higher oxidation-reduction potential under the same effective chlorine concentration. Therefore, the disinfection and sterilization liquid according to one embodiment of the invention has better disinfection capability and disinfection speed, and has no toxicity and irritation to organisms. Furthermore, the disinfectant sterilizing liquid of one embodiment of the invention can be used for preparing a composition for treating wounds or treating infection. Treating the wound may include disinfecting the wound resulting from the surgical procedure, the decubitus ulcer, or the burn and scald. Treating the infection may comprise disinfecting infection caused by surgery, peritonitis, bedsores, or burns, and may comprise disinfecting infection caused by bacteria or viruses. By performing wound treatment or infection treatment using the disinfectant sterilizing liquid according to one embodiment of the present invention, infection can be prevented or inhibited, thereby accelerating wound healing.

Description of the symbols

None.

Claims

1. A disinfection solution comprises sodium ions, a first weak acid, a second weak acid and water;

wherein the first weak acid and the second weak acid are different weak acids, and the second weak acid is hypochlorous acid;

the effective chlorine concentration of the disinfection and sterilization liquid is more than 25ppm and less than or equal to 200ppm, and the effective chlorine of the disinfection and sterilization liquid is only from hypochlorous acid; and is

The oxidation-reduction potential of the disinfection and sterilization liquid is 1100 to 1500 millivolts.

2. A disinfecting and sterilizing solution, basically comprising sodium ions, a first weak acid, a second weak acid and water;

the effective chlorine concentration of the disinfection and sterilization liquid is more than 25ppm and less than or equal to 200 ppm; and is

3. The disinfectant sterilizing solution according to claim 1 or 2, wherein the first weak acid comprises citric acid, carbonic acid or acetic acid.

4. The disinfectant sterilizing liquid according to claim 1 or 2, wherein the disinfectant sterilizing liquid has an effective chlorine concentration of more than 100ppm and 200ppm or less.

5. The disinfectant sterilizing liquid according to claim 4, wherein the oxidation-reduction potential of the disinfectant sterilizing liquid is 1300 to 1500 millivolts.

6. The disinfectant sterilizing liquid according to claim 1 or 2, wherein the pH of the disinfectant sterilizing liquid is 4 to 6.

7. The disinfectant sterilizing solution according to claim 1 or 2, wherein the pKa of the first weak acid is 4 to 7.

8. A method of making a disinfecting solution of any one of claims 1-7 comprising mixing sodium hypochlorite, a first weak acid and water to react to form a disinfecting solution;

wherein the disinfection sterilization liquid comprises sodium ions, the first weak acid, the second weak acid and water;

the first weak acid and the second weak acid are different weak acids, and the second weak acid is hypochlorous acid;

9. The method of claim 8, wherein the weight ratio of the sodium hypochlorite to the first weak acid is 1.0:0.5 to 1.0: 1.5.

10. The method of claim 8 or 9, wherein the first weak acid comprises citric acid, carbonic acid, or acetic acid.

11. Use of a disinfecting and sterilizing solution for the preparation of a composition for treating a wound, wherein the disinfecting and sterilizing solution is as defined in any one of claims 1 to 7.

12. The use of claim 11, wherein treating a wound comprises disinfecting a wound caused by surgery, pressure sores, or burns.

13. Use of a disinfectant and bactericidal solution for the preparation of a composition for treating infections, wherein the disinfectant and bactericidal solution is as defined in any one of claims 1 to 7.

14. The use of claim 13, wherein treating the infection comprises disinfecting the infection caused by surgery, peritonitis, bedsores, or burns.

15. The use of claim 13 or 14, wherein treating an infection comprises treating an infection caused by a drug-resistant staphylococcus aureus, acinetobacter baumannii, bacillus subtilis, clostridium, staphylococcus aureus, escherichia coli, pseudomonas aeruginosa, candida albicans, salmonella enterica, streptococcus mutans, klebsiella pneumoniae, propionibacterium acnes, aspergillus niger, listeria, enterovirus, coronavirus, or influenza virus.