PHMB disinfectant neutralizer as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of neutralizers for disinfectants, and particularly relates to a PHMB disinfectant neutralizer as well as a preparation method and application thereof. The neutralizer is prepared from the following components in percentage by weight: 0.3%-0.5% of Tween-80, 0.5%-0.75% of lecithin, 0.15%-0.3% of calcium stearate, 0.15%-0.3% of sodium dodecyl benzene sulfonate and the balance of water. The neutralizer provided by the invention is suitable for laboratory disinfection experiments to evaluate the sterilization effect of guanidine disinfection products, and has the following advantages: the corresponding guanidine disinfection products can be neutralized more quickly and more effectively; the neutralizer and a neutralization product generated by the reaction of the neutralizer and the guanidine disinfection product do not have any effect of killing or inhibiting the growth of experimental microorganisms; no damage is caused to culture components, and no influence is caused to the characters of the culture components.

Description

Technical Field

[0001] The invention belongs to the technical field of neutralizers for disinfectants, and in particular relates to a PHMB disinfectant neutralizer and a preparation method and application thereof. Background Art

[0002] The purpose of the bactericidal effect evaluation experiment of disinfectants is to accurately test their bactericidal, bacteriostatic and antibacterial effects. In order to ensure the high accuracy of the test results, the action time of the disinfectant must be precisely controlled, and its disinfection effect must be quickly terminated after the predetermined time is reached to avoid the continued influence of the disinfectant during the recovery culture of microorganisms. Therefore, selecting a neutralizer that can effectively block the bactericidal effect of the disinfectant is extremely critical for accurately evaluating the bactericidal effect of disinfectant products. If effective measures are not taken to remove residual disinfectants during the experiment, it will be regarded as a major mistake in the experimental design and the results obtained will be unreliable.

[0003] Guanidine polymers are named because of the alkylguanidines contained in their chemical structure. They are a class of biologically active organic bases with a very wide range of applications. As early as the 1930s, people had discovered its bactericidal ability and continued to research and develop it. Polyhexamethylene biguanide hydrochloride (PHMB) is recognized as the safest and most effective broad-spectrum antibacterial agent in the 21st century. It is colorless and odorless, has a low antibacterial concentration, a fast action rate, a low foam volume, and can form a cationic film on the surface of objects to continuously inhibit bacteria for a long time without generating antibacterial bacteria. It is widely used in various fields such as medical devices, public environments, homes, fabrics, food, milk, and care products, and has become one of the most commonly used disinfectants in people's daily lives.

[0004] The bactericidal mechanism of PHMB is that the structure of its high molecular polymer makes the guanidine group highly active, which can make the polymer positively charged and easily adsorbed by various negatively charged bacteria and viruses, thereby inhibiting the division function of bacteria and viruses and making them lose their reproductive ability. In addition, the film formed by the polymer blocks the respiratory tract of microorganisms, causing microorganisms to suffocate and die quickly. This feature makes it have a broad-spectrum bactericidal power, especially for Gram-negative bacteria that are difficult to treat with general bactericides. However, there are different evaluations of its bactericidal performance in the academic community. Some scholars believe that guanidine disinfectants themselves have strong bactericidal ability, while others believe that this is due to the fact that guanidine disinfectants have not been able to find a suitable neutralizer.

[0005] At present, the laboratory has existing neutralizer formulas for PHMB disinfectants, most of which are combinations of compounds such as lecithin, Tween-80, sodium thiosulfate, and histidine. Although this type of neutralizer can play a certain neutralizing role on PHMB, this type of neutralizer also often has some problems. Either the results of the neutralization identification experiment are poorly repeatable, which limits its scope of application; or the neutralization effect is not satisfactory and it is difficult to meet the use requirements, which is very unfavorable for testing the sterilization performance of guanidine disinfectants. Summary of the invention

[0006] The present invention aims to introduce a novel neutralizing agent to solve many deficiencies of existing guanidine disinfectant neutralizing agents.

[0007] The first object of the present invention is to provide a PHMB disinfectant neutralizer for the above-mentioned problem, wherein the neutralizer is made of the following components by weight percentage:

[0008] Tween-80 0.3%-0.5%, lecithin 0.5%-0.75%, calcium stearate 0.15%-0.3%, sodium dodecylbenzene sulfonate 0.15%-0.3%, and the balance is water.

[0009] The neutralizer of the present invention is used for neutralization of disinfectants in microbial effect verification including quantitative bacterial killing tests, so as to overcome the defects of existing neutralizers, and can more quickly and effectively neutralize corresponding guanidine disinfection products. The neutralizer itself and its neutralized product have no killing or growth inhibition effect on experimental microorganisms, do not cause any damage to culture components, and do not have any effect on their physical properties. In addition, the components of the neutralizer are simple, easy to make, and can be well applied to various sterilization tests.

[0010] The second object of the present invention is to provide a method for preparing the neutralizing agent, comprising the following steps:

[0011] 1) weighing calcium stearate and sodium dodecylbenzene sulfonate in proportion, heating and dissolving them in water to obtain solution A;

[0012] 2) Tween-80 and lecithin are weighed in proportion, heated and dissolved in water, and then cooled to obtain solution B;

[0013] 3) Solution A and solution B are mixed evenly, and the neutralizing agent is obtained after cooling, fixing the volume and sterilizing.

[0014] As a preferred embodiment of the preparation method of the present invention, in step 1), calcium stearate and sodium dodecylbenzene sulfonate are added to water, heated to 20-27° C., and stirred to dissolve the calcium stearate and sodium dodecylbenzene sulfonate.

[0015] As a preferred embodiment of the preparation method of the present invention, in step 2), Tween-80 and lecithin are added to water, heated to 54-55° C., and stirred to dissolve the Tween-80 and lecithin.

[0016] As a preferred embodiment of the preparation method of the present invention, in step 3), high temperature and high pressure sterilization is adopted, and high pressure sterilization is carried out at 120-121° C. and 0.2 MPa for 15 minutes.

[0017] The second object of the present invention is to provide the use of the PHMB disinfectant neutralizer in evaluating the bactericidal effect of guanidine disinfection products.

[0018] In a specific embodiment of the present invention, the guanidine disinfection product includes PHMB disinfectant.

[0019] In a specific embodiment of the present invention, the bacteria include Escherichia coli and Candida albicans.

[0020] The principle of the neutralizing agent of the present invention to neutralize the PHMB disinfectant is as follows: the PHMB disinfectant is a high molecular polymer with an alkylguanidine active group and exhibits positive charge, while Tween-80, as a nonionic surfactant and emulsifier, has a strong dispersing ability, can remove the disinfectant adsorbed on the surface of the bacteria and block the combination of the disinfectant and the bacteria, lecithin is a biosurfactant with a phosphate group as a hydrophilic group, has an interface adsorption ability, can form a film on the surface of the bacteria to block the contact with the disinfectant; calcium stearate and sodium dodecylbenzene sulfonate are both anionic Surfactant, calcium stearate can hydrolyze calcium ions and negatively charged stearate ions, sodium dodecylbenzene sulfonate generates sulfonate after ionization in water, has good degreasing ability and reduces the surface tension of water and the properties of wetting, penetration and emulsification. In addition, sodium dodecylbenzene sulfonate is also easy to produce synergistic effects with other non-ionic surfactant substances. When these four components are combined with the positively charged PHMB disinfectant, they can react with it to form an insoluble flocculent complex and block the contact between the microbial surface and PHMB, thereby neutralizing its disinfection effect.

[0021] The present invention has the following beneficial effects:

[0022] First, the neutralizer of the present invention can neutralize the corresponding guanidine disinfection products more quickly and effectively; second, the neutralizer itself and the neutralization product produced by the reaction with the guanidine disinfection product have no effect on killing or inhibiting the growth of experimental microorganisms; finally, the neutralizer does not cause any damage to the culture components and will not have any effect on their physical properties. DETAILED DESCRIPTION

[0023] The present invention is described in detail below in conjunction with specific examples, but it should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0024] Example 1

[0025] The present embodiment provides a PHMB disinfectant neutralizer, which is made of the following components by weight percentage: 0.5% lecithin, 0.3% Tween-80, 0.15% calcium stearate, 0.15% sodium dodecylbenzene sulfonate, and 98.9% purified double distilled water.

[0026] The preparation method of the PHMB disinfectant neutralizer of the present embodiment comprises the following steps:

[0027] 1) Accurately measure 1.5 g of calcium stearate and sodium dodecylbenzene sulfonate according to the ratio and dissolve them in purified double distilled water, heat to 25° C., and stir to fully dissolve them to obtain solution A;

[0028] 2) Accurately measure 3 g of Tween-80 and 5 g of lecithin according to the ratio and dissolve them in purified double distilled water, heat to 54° C., and stir to fully dissolve them to obtain solution B;

[0029] 3) Mix solution A and solution B according to the formula ratio and stir evenly, cool and dilute to 1 kg before packaging;

[0030] 4) The solution obtained in step 3) is sterilized by high pressure sterilization at 121° C. and 0.2 MPa for 15 min, and is put into storage after passing the sterility test, thereby obtaining the neutralizing agent.

[0031] Example 2

[0032] The present embodiment provides a PHMB disinfectant neutralizer, which is made of the following components by weight percentage: 0.6% lecithin, 0.4% Tween-80, 0.2% calcium stearate, 0.2% sodium dodecylbenzene sulfonate, and 98.6% purified double distilled water.

[0033] The preparation method of the PHMB disinfectant neutralizer of the present embodiment comprises the following steps:

[0034] 1) Accurately measure 2 g of calcium stearate and sodium dodecylbenzene sulfonate according to the ratio and dissolve them in purified double distilled water, heat to 25°C, and stir to fully dissolve them to obtain solution A;

[0035] 2) Accurately measure 4 g of Tween-80 and 6 g of lecithin according to the ratio and dissolve them in purified double distilled water, heat to 54° C., and stir to fully dissolve them to obtain solution B;

[0036] 3) Mix solution A and solution B according to the formula ratio and stir evenly, and then package after cooling and fixing the volume;

[0037] 4) The solution obtained in step 3) is sterilized by high pressure sterilization at 121° C. and 0.2 MPa for 15 min, and is put into storage after passing the sterility test, thereby obtaining the neutralizing agent.

[0038] Example 3

[0039] The present embodiment provides a PHMB disinfectant neutralizer, which is made of the following components by weight percentage: 0.75% lecithin, 0.5% Tween-80, 0.3% calcium stearate, 0.3% sodium dodecylbenzene sulfonate, and 98.6% purified double distilled water.

[0040] The preparation method of the PHMB disinfectant neutralizer of the present embodiment comprises the following steps:

[0041] 1) Accurately measure 3 g of calcium stearate and sodium dodecylbenzene sulfonate according to the ratio and dissolve them in purified double distilled water, heat to 25°C, and stir to fully dissolve them to obtain solution A;

[0042] 2) Accurately measure 5 g of Tween-80 and 7.5 g of lecithin according to the ratio and dissolve them in purified double distilled water, heat to 54° C., and stir to fully dissolve them to obtain solution B;

[0043] 3) Mix solution A and solution B according to the formula ratio and stir evenly, and then package after cooling and fixing the volume;

[0044] 4) The solution obtained in step 3) is sterilized by high pressure sterilization at 121° C. and 0.2 MPa for 15 min, and is put into storage after passing the sterility test, thereby obtaining the neutralizing agent.

[0045] The effects of Example 1, Example 2 and Example 3 are similar. In order to more clearly and in detail introduce the effect of the neutralizer described in the embodiment of the present invention, the present invention will be further described below in conjunction with Example 1.

[0046] Neutralizer effect test, the method is as follows:

[0047] According to the requirements of GB / T38502-2020 "Test Methods for Laboratory Bactericidal Effect of Disinfectants", 6 groups of experiments were set up, each group was repeated 3 times, and the horizontal effects of different neutralizers were compared to determine the selection advantage of the selected neutralizer. The test results require that Group 1 should not grow bacteria or have a few colonies growing; Group 2 should have more colonies than Group 1, but significantly less than Groups 3, 4, and 5; Groups 3, 4, and 5 should have similar bacterial growth and the number of colonies should be between 1×10 5 ~5×10 5However, the error rate among the three groups should not exceed 15%; and the colony error rate among the groups should not exceed 15%, which means that the composition and concentration of the test neutralizer are appropriate and can effectively neutralize the target disinfectant. The experimental groups are as follows:

[0048] Table 1 Test groups

[0049]

[0050] Choose the common PHMB disinfectant on the market with a mass concentration of 20%, dilute it to 0.3% with purified double distilled water, and refrigerate it to 4°C in a refrigerator for later use.

[0051] The strains used in the test were Escherichia coli 25922 and Candida albicans ATCC 10231. The diluent used was tryptone physiological saline solution.

[0052] Freeze-dried bacterial tube, add appropriate amount of nutrient broth medium to melt and disperse the bacterial strain. Pipette a small amount of bacterial strain into a test tube with appropriate amount of nutrient broth medium, and incubate at 36±1℃ for an appropriate period of time to make bacterial solution. Use OD 600 The absorbance measured by spectrophotometer was about 0.3, and the bacterial content of the bacterial solution was 1.0×10 6 ~3×10 6 CFU / ml, and then mixed evenly with bovine serum albumin BSA solution with a mass concentration of 0.3% in a volume ratio of 1:1 to obtain a bacterial suspension.

[0053] The above disinfectant was used to conduct a neutralization identification test, the neutralizer described in Specific Example 1 was used as the test group neutralizer, and five neutralizers were set as comparative examples for testing experiments, wherein the neutralizers of the five comparative examples were:

[0054] The neutralizing agent of Comparative Example 1 is made of the following components calculated by mass percentage: 0.3% Tween-80, 0.5% lecithin, and the balance is purified water;

[0055] The neutralizer of Comparative Example 2 is made of the following components calculated by mass percentage: 0.3% Tween-80 and the remainder D / E neutralizing broth.

[0056] The neutralizer of Comparative Example 3 is made of the following components calculated by mass percentage: 0.3% Tween-80, 0.5% lecithin, 0.5% histidine, 0.5% sodium thiosulfate, and the balance is distilled water.

[0057] The neutralizer of Comparative Example 4 is made of the following components calculated by mass percentage: 3% Tween-80, 0.3% lecithin, 0.3% soap base, and the balance PBS phosphate buffer solution.

[0058] The neutralizer of Comparative Example 5 is made of the following components calculated by mass percentage: 0.25% Tween-80, 0.3% lecithin, 0.25% sodium thiosulfate, 0.05% sodium sulfite, and the balance is PBS phosphate buffer solution.

[0059] The results of the neutralization identification test of each neutralizer on 0.3% PHMB disinfectant are shown in Table 2 and Table 3:

[0060] Table 2 Results of the neutralization and identification test of Escherichia coli of PHMB disinfectant

[0061]

[0062] Note: The temperature is room temperature, 0.3% PHMB acts on E. coli for 10 seconds, and the result is the average of 3 tests.

[0063] Table 3 Candida albicans neutralization identification test results of PHMB disinfectant

[0064]

[0065]

[0066] Note: The temperature is room temperature, 0.3% PHMB acts on Candida albicans for 10 seconds, and the results are the average of 3 tests.

[0067] The experimental results in Table 2 and Table 3 show that the colony error rates of the neutralizers in the experimental group and the comparative example for the PHMB disinfectant samples are no more than 15%, and the neutralization effects on the PHMB disinfectant reflect that the neutralizer of the present invention is superior to the five neutralizers in the comparative example, that is, the neutralizer of the present invention can completely and efficiently neutralize the PHMB disinfectant, can effectively avoid the influence of the disinfectant residue on the accuracy of the bactericidal effect verification test, and can meet the requirements of national statutory regulations.

[0068] It should be noted that when the claims of the present invention involve numerical ranges, it should be understood that the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. In order to avoid redundancy, the present invention describes a preferred embodiment.

[0069] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

Claims

1. A PHMB disinfectant neutralizer, characterized in that: The neutralizing agent is made of the following components by weight percentage: Tween-80 0.3%-0.5%, lecithin 0.5%-0.75%, calcium stearate 0.15%-0.3%, sodium dodecylbenzene sulfonate 0.15%-0.3%, and the balance is water.

2. The method for preparing the PHMB disinfectant neutralizer according to claim 1, characterized in that: The following steps are involved: 1) weighing calcium stearate and sodium dodecylbenzene sulfonate in proportion, heating and dissolving them in water to obtain solution A; 2) Tween-80 and lecithin are weighed in proportion, heated and dissolved in water, and then cooled to obtain solution B; 3) Solution A and solution B are mixed, and the neutralizing agent is obtained after cooling, fixing the volume and sterilizing.

3. The preparation method according to claim 2, characterized in that: In step 1), calcium stearate and sodium dodecylbenzene sulfonate are added into water, heated to 20-27° C., and stirred to dissolve calcium stearate and sodium dodecylbenzene sulfonate.

4. The preparation method according to claim 2, characterized in that: In step 2), Tween-80 and lecithin are added to water, heated to 54-55° C., and stirred to dissolve the Tween-80 and lecithin.

5. The preparation method according to claim 2, characterized in that: In step 3), high temperature and high pressure sterilization is adopted, and high pressure sterilization is carried out at 120-121° C. and 0.2 MPa for 15 minutes.

6. Use of the PHMB disinfectant neutralizer according to claim 1 in evaluating the bactericidal effect of guanidine disinfection products.

7. The use according to claim 6, characterized in that: The guanidine disinfectant products include PHMB disinfectant.

8. The use according to claim 6, characterized in that: The bacteria include Escherichia coli and Candida albicans.

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