阅读说明：本技术 新生儿脐部护理液及其制备方法 (Newborn umbilical region care solution and preparation method thereof ) 是由 刘果 秦翠林 于 2021-07-30 设计创作，主要内容包括：本申请提供一种新生儿脐部护理液,包括：液态溶剂,重量比为96-99份；第一溶质,包括石墨烯；第二溶质,包括聚六亚甲基双胍盐酸盐；以及分散剂,用于协助所述第一溶质及所述第二溶质在所述液态溶剂中分散,重量比为0.4至1.6份。本申请实施例还提供一种新生儿脐部护理液的制备方法,包括如下步骤：将第二溶质溶于液态溶剂,制备第一混合液,所述第二溶质包括聚六亚甲基双胍盐酸盐；将第一溶质及分散剂分别溶于所述第一混合液,制备第二混合液,所述第一溶质包括石墨烯,所述分散剂用于协助所述第一溶质及所述第二溶质在所述液态溶剂中分散；对所述第二混合液进行分散处理得到新生儿脐部护理液。(The application provides a neonate umbilical region nursing liquid includes: 96-99 parts of liquid solvent; a first solute comprising graphene; a second solute comprising polyhexamethylene biguanide hydrochloride; and a dispersing agent for assisting dispersion of the first solute and the second solute in the liquid solvent, in a weight ratio of 0.4 to 1.6 parts. The embodiment of the application also provides a preparation method of the newborn umbilical region care solution, which comprises the following steps: dissolving a second solute in a liquid solvent to prepare a first mixed solution, wherein the second solute comprises polyhexamethylene biguanide hydrochloride; respectively dissolving a first solute and a dispersant in the first mixed solution to prepare a second mixed solution, wherein the first solute comprises graphene, and the dispersant is used for assisting the first solute and the second solute in dispersing in the liquid solvent; and performing dispersion treatment on the second mixed solution to obtain the newborn umbilical nursing solution.)

1. A newborn umbilical region nursing liquid is characterized by comprising:

96-99 parts of liquid solvent;

a first solute comprising graphene;

a second solute comprising polyhexamethylene biguanide hydrochloride; and

a dispersant for assisting dispersion of the first solute and the second solute in the liquid solvent in a weight ratio of 0.4 to 1.6 parts.

2. The neonatal umbilical care solution of claim 1, wherein the graphene comprises nano graphene or graphene oxide.

3. The newborn umbilical care solution according to claim 1, wherein the weight ratio of the graphene is 0.005 to 0.008 parts.

4. The umbilical cord care solution of claim 1 wherein the polyhexamethylene biguanide hydrochloride is present in an amount of 0.05 to 0.1 parts by weight.

5. The newborn navel nursing solution according to claim 1, wherein the dispersing agent comprises at least one of sodium dodecyl benzene sulfonate, polyvinyl alcohol, polyacrylamide, polyethylene glycol, polyethylene oxide and polyvinylpyrrolidone.

6. The neonatal umbilical care solution of claim 1, wherein the liquid solvent is purified water or deionized water.

7. A method for preparing the umbilical cord of the newborn infant care solution as claimed in any one of claims 1 to 6, comprising the steps of:

dissolving a second solute in a liquid solvent to prepare a first mixed solution, wherein the second solute comprises polyhexamethylene biguanide hydrochloride;

respectively dissolving a first solute and a dispersant in the first mixed solution to prepare a second mixed solution, wherein the first solute comprises graphene, and the dispersant is used for assisting the first solute and the second solute in dispersing in the liquid solvent; and

and performing dispersion treatment on the second mixed solution to obtain the newborn umbilical nursing solution.

8. The method of preparing the newborn umbilical care solution according to claim 7, wherein the graphene comprises nano graphene or graphene oxide, the weight ratio of the graphene is 0.005 to 0.008 parts, and the weight ratio of the polyhexamethylene biguanide hydrochloride is 0.05 to 0.1 parts.

9. The method of claim 7, wherein the dispersant comprises at least one of sodium dodecylbenzenesulfonate, polyvinyl alcohol, polyacrylamide, polyethylene glycol, polyethylene oxide, and polyvinylpyrrolidone, and the liquid solvent is purified water or deionized water.

10. The method of preparing a solution for umbilical cord of the newborn according to claim 7, wherein the method of dispersing the second mixed solution includes ultrasonic dispersion.

Technical Field

The application relates to the technical field of biological medicines, in particular to a neonate umbilical region care solution and a preparation method thereof.

Background

The umbilical cord is the passage for the fetus between the mother and the mother to supply the fetus with nutrients and waste products. After the fetus comes out, the medical staff ligates and cuts off the umbilical cord, and then the umbilical cord stump gradually becomes dry and thin and falls off after 3-7 days after the baby comes out. However, before and after umbilical cord amputation, if nursing is not performed properly, bacterial contamination is easily caused, and inflammation is caused to the umbilical part of the newborn.

The main products in the market at present mainly used for nursing the umbilical region of the newborn are as follows: (1) alcohols (e.g., 75% alcohol, polyethylene glycol, etc.); (2) iodine (such as iodine tincture, iodophor); dressings and the like. However, the above-mentioned types of products all have drawbacks when sterilizing the umbilical region of a newborn: (1) 75% alcohol is mainly used for intact skin (such as skin disinfection before injection, puncture or operation), and can not be used in skin damage and erosion and effusion; and can be irritating to the wound. (2) Iodine tincture is used for sterilizing the whole skin, and 70% ethanol is required to be used for deiodination after 1 minute; large area skin defects are unusable and can damage granulation tissue. Iodophor is used for skin disinfection, mucous membrane flushing, hand brushing, hand soaking, injection and skin disinfection of operation positions of medical staff and treatment of skin mucous membrane bacterial infection, but has the problem of cautious use for people with iodine or povidone iodine allergy, and the Chinese pharmacopoeia determines that visible iodine is obviously absorbed by skin for external use of infants, and the iodophor should be cautious to infants. (3) Medical dressings: the main components of the liquid dressing are high molecular materials such as carbomer, polyvinyl alcohol, sodium hyaluronate and the like, the liquid dressing acts on wounds to form a film, the ventilation and the drying of the wounds are not facilitated, and the formed product has multiple technical components and is easy to cause irritation and allergy; the wound surface dressing usually consists of a gluing base material, an absorptive dressing pad and a strippable protective layer, the dressing has poor air permeability, needs to be fixed by adhesive, and the fixed part is easy to cause allergy, red, swollen, itchy and other adverse reactions.

How to solve the above problems needs to be considered by those skilled in the art.

Disclosure of Invention

The embodiment of the application provides a neonate umbilical part nursing liquid, includes:

96-99 parts of liquid solvent;

a first solute comprising graphene;

a second solute comprising polyhexamethylene biguanide hydrochloride; and

a dispersant for assisting dispersion of the first solute and the second solute in the liquid solvent in a weight ratio of 0.4 to 1.6 parts.

In one possible embodiment, the graphene comprises nano-graphene or graphene oxide.

In one possible embodiment, the weight ratio of the graphene is 0.005 to 0.008 parts.

In one possible embodiment, the polyhexamethylene biguanide hydrochloride is present in a weight ratio ranging from 0.05 to 0.1 parts.

In one possible embodiment, the dispersant comprises at least one of sodium dodecylbenzene sulfonate, polyvinyl alcohol, polyacrylamide, polyethylene glycol, polyethylene oxide, polyvinylpyrrolidone.

In one possible embodiment, the liquid solvent is purified water or deionized water.

The embodiment of the application also provides a preparation method of the newborn umbilical region care solution, which comprises the following steps:

dissolving a second solute in a liquid solvent to prepare a first mixed solution, wherein the second solute comprises polyhexamethylene biguanide hydrochloride;

respectively dissolving a first solute and a dispersant in the first mixed solution to prepare a second mixed solution, wherein the first solute comprises graphene, and the dispersant is used for assisting the first solute and the second solute in dispersing in the liquid solvent; and

and performing dispersion treatment on the second mixed solution to obtain the newborn umbilical nursing solution.

In one possible embodiment, the graphene includes nano-graphene or graphene oxide, the weight ratio of the graphene is 0.005 to 0.008 parts, and the weight ratio of the polyhexamethylene biguanide hydrochloride is 0.05 to 0.1 parts.

In one possible embodiment, the dispersant comprises at least one of sodium dodecylbenzene sulfonate, polyvinyl alcohol, polyacrylamide, polyethylene glycol, polyethylene oxide, and polyvinylpyrrolidone, and the liquid solvent is purified water or deionized water.

In one possible embodiment, the method of performing the dispersion treatment on the second mixed liquid includes ultrasonic dispersion.

Compared with the prior art, the newborn umbilical region care solution and the preparation method thereof are provided. On one hand, the bactericidal capacity of the graphene and the derivatives thereof mainly comes from the physical destruction effect, bacteria hardly generate resistance to the graphene and the derivatives thereof, and the antibacterial material of the graphene and the derivatives thereof has the characteristics of high action efficiency, small using amount, easy modification, simple preparation process, capability of effectively reducing the biotoxicity, good biocompatibility and the like. On the other hand, the polyhexamethylene biguanide hydrochloride has a certain bactericidal effect, and can be synergistically acted with the graphene and the derivatives thereof, so that the bactericidal effect is more obvious.

Drawings

Fig. 1 is a schematic flow chart of a preparation process of a neonate umbilical region care solution according to an embodiment of the present application.

Fig. 2 is a schematic diagram of an experimental electron microscope according to an embodiment of the present application.

The following detailed description will further illustrate the present application in conjunction with the above-described figures.

Detailed Description

The following description will refer to the accompanying drawings to more fully describe the present disclosure. There is shown in the drawings exemplary embodiments of the present application. This application may, however, be embodied in many different forms and should not be construed as limited to the exemplary embodiments set forth herein. These exemplary embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art. Like reference numerals designate identical or similar components.

The terminology used herein is for the purpose of describing particular example embodiments only and is not intended to be limiting of the application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. Further, as used herein, "comprises" and/or "comprising" and/or "having," integers, steps, operations, components, and/or components, but does not preclude the presence or addition of one or more other features, regions, integers, steps, operations, components, and/or groups thereof.

Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Furthermore, unless otherwise defined herein, terms such as those defined in commonly used dictionaries should be interpreted as having a meaning that is consistent with their meaning in the context of the relevant art and the present application and will not be interpreted in an idealized or overly formal sense.

The following description of exemplary embodiments refers to the accompanying drawings. It should be noted that the components depicted in the referenced drawings are not necessarily shown to scale; and the same or similar components will be given the same or similar reference numerals or similar terms.

Embodiments of the present application will now be described in further detail with reference to the accompanying drawings.

The embodiment of the application provides a neonate umbilical region care solution, including liquid solvent, first solute, second solute and dispersant are dissolved in the liquid solvent.

In one embodiment, the liquid solvent is purified water or deionized water in a weight ratio of 96-99 parts.

In one embodiment, the first solute includes graphene, the graphene includes nano-graphene or graphene oxide, and the weight ratio of the graphene is 0.005 to 0.008 parts. The newborn umbilical region care solution with the proportion has a strong sterilization effect, and is shown as follows:

1. staphylococcus aureus (ATTCC6538), Escherichia coli (ATCC25922) and Candida albicans (ATCC10231) were respectively prepared into a bacterial solution with a bacterial content of 1 × 108 CFU/ml.

2. Weighing a certain amount of graphene or graphene oxide, and preparing into water solutions with mass concentrations of 1, 2, 3, 5 and 8 mg/ml.

3. 100ul of the above-mentioned bacterial solutions were taken, respectively. And (3) adding 1ml of the graphene or graphene oxide aqueous solution with each concentration into each bacterial liquid to be tested, and uniformly mixing. Blank control is to add equal volume of distilled water to each bacterial solution to mix well. Taking 100ul of the mixed solution, coating the mixed solution on an LB agar culture medium, culturing for 24h in an incubator at 37 ℃, and counting the number of colonies on a plate.

4. The method for calculating the bacteriostasis rate comprises the following steps:

the bacteriostasis rate is (number of blank control colonies-number of sample colonies)/number of blank control colonies multiplied by 100%

5. The results are as follows:

the results are shown in the table above, when the mass concentration of the graphene or the graphene oxide is 1-3mg/ml, the bacteriostasis rate of each test strain of the graphene aqueous solution is 30.55-73.94% of that of staphylococcus aureus respectively; 40.23-70.12% of escherichia coli; 47.56-70.64% of candida albicans; the bacteriostasis rate of each test strain of the graphene oxide aqueous solution is 32.48-76.89% of staphylococcus aureus; 45.56-75.19% of Escherichia coli; candida albicans 49.23-72.11%. And when the mass concentration of the graphene aqueous solution and the graphene oxide aqueous solution is 5-8mg/ml, the sterilization rate of each test strain reaches 99.99%. Exhibits high bactericidal action.

It can be understood that when the mass concentration of the graphene aqueous solution or the graphene oxide aqueous solution reaches a certain value, cytotoxicity may be caused. As shown in fig. 2, when the mass concentration of graphene is 3mg/ml, it can be seen that the cell state is good, the cell and graphene are well adhered, and phagocytosis and wrapping phenomena occur on part of graphene; when the concentration is 8mg/ml, the cell body becomes small, the surface secretion is obviously increased, and the microvilli on the cell surface becomes long, which indicates that the cells are stimulated; when the concentration is 9mg/ml, cells shrink and deform, most of the cells are inactivated and die, and the graphene has high cytotoxicity to the cells.

In other embodiments, the first solute may further include other graphene-based derivatives, and the weight ratio of the graphene may be 0.5 to 1 part, 1 to 1.5 parts, and 1.5 to 2 parts.

In one embodiment, the second solute comprises polyhexamethylene biguanide hydrochloride in a weight ratio of 0.05 to 0.1 parts.

The polyhexamethylene biguanide hydrochloride has a certain bactericidal effect, and can be synergistically acted with the graphene and the derivatives thereof, so that the bactericidal effect is more obvious.

Polyhexamethylene biguanide hydrochloride (PHMB) is a broad-spectrum antibiotic, has killing effects on gram-positive bacteria, gram-negative bacteria, fungi, yeasts and the like, and can effectively kill various bacteria and viruses such as staphylococcus aureus, escherichia coli, dysentery bacillus, candida albicans and the like. And can achieve good sterilization effect at low concentration, and meanwhile, can not cause bacteria to generate drug resistance. The polyhexamethylene biguanide hydrochloride is an active antibacterial agent of cell membranes, and the antibacterial mechanism is as follows: firstly, polyhexamethylene biguanide hydrochloride is a polycationic compound and can be adsorbed on the surface of a negatively charged microorganism to prevent the microorganism from dividing and propagating; meanwhile, the polyhexamethylene biguanide hydrochloride adsorbed on the surface of the microorganism acts on a phospholipid bilayer in cytoplasm through transmembrane transport and is combined with a phosphate group with negative electricity, so that the selective permeability of a cell membrane is changed, the cell metabolism is damaged, and the microorganism is killed.

By utilizing the positive charge characteristic of the polyhexamethylene biguanide hydrochloride, the surface of the graphene can carry cationic groups after the polyhexamethylene biguanide hydrochloride is compounded with the graphene, and the electrostatic adsorption effect of the graphene on bacteria is enhanced. Meanwhile, the compounded graphene has a larger surface area, so that the graphene is more easily contacted with bacteria, the integral death rate of the bacteria is increased, the sterilization effect is stronger, and the synergistic effect is generated. Meanwhile, the PHMB cations are adsorbed on the surface of the object after sterilization, so that the growth of bacteria can be inhibited for a long time.

Confirmation of antibacterial long-lasting property:

1. the method comprises the following steps: 5 pieces of disc-shaped fabric with the diameter of 3cm are overlapped, a sample is sprayed on the fabric to completely soak the fabric, and the antibacterial durability simulation evaluation is carried out after the sample is subjected to 50 ℃ durability simulation heat treatment for 16 hours and ultraviolet lamp irradiation for 100 hours of light resistance treatment according to an antibacterial processing product antibacterial force durability test method provided by the Japan antibacterial product Association.

2. Staphylococcus aureus and Escherichia coli were used as test strains.

3. As a result: the samples with added biocide (PHMB) still showed good antimicrobial effect after heat and light experiments compared to the samples without the added biocide.

In one embodiment, the dispersant comprises at least one of sodium dodecylbenzene sulfonate, polyvinyl alcohol, polyacrylamide, polyethylene glycol, polyethylene oxide, and polyvinylpyrrolidone.

The dispersing agent is used for assisting the dispersion of the first solute and the second solute in the liquid solvent, and the weight ratio is 0.4-1.6 parts. The dispersing agent can improve the dispersing effect of the graphene in the liquid solvent, so that the effect of the graphene for sterilization is better.

As shown in fig. 1, an embodiment of the present application further provides a preparation method of a neonate umbilical region care solution, including the following steps:

step S1: and dissolving a second solute in the liquid solvent to prepare a first mixed solution, wherein the second solute comprises polyhexamethylene biguanide hydrochloride.

In one embodiment, the polyhexamethylene biguanide hydrochloride is present in an amount of 0.05 to 0.1 parts by weight.

Step S2: respectively dissolving a first solute and a dispersant in the first mixed solution to prepare a second mixed solution, wherein the first solute comprises graphene, and the dispersant is used for assisting the first solute and the second solute in dispersing in the liquid solvent.

In one embodiment, the graphene includes nano-graphene or graphene oxide, and the weight ratio of the graphene is 0.005 to 0.008 parts.

In one embodiment, the dispersant includes at least one of sodium dodecylbenzene sulfonate, polyvinyl alcohol, polyacrylamide, polyethylene glycol, polyethylene oxide, and polyvinylpyrrolidone, and the liquid solvent is purified water or deionized water.

Step S3: and performing dispersion treatment on the second mixed solution to obtain the newborn umbilical nursing solution.

In an embodiment, the method of performing the dispersion treatment on the second mixture includes ultrasonic dispersion.

Example 1

Graphene oxide	0.05
		Polyvinylpyrrolidone	0.5
Polyhexamethylene biguanide hydrochloride	0.1
		Purified water	99.0

Adding polyvinylpyrrolidone into a container filled with polyhexamethylene biguanide hydrochloride aqueous solution, stirring to disperse and dissolve, adding graphene oxide, performing ultrasonic treatment to disperse uniformly, and subpackaging in a proper container to obtain the care solution.

And (3) carrying out effect test on the prepared care solution:

quantitative bacteria killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; staphylococcus aureus (ATTCC6538), Pseudomonas aeruginosa (ATCC15442) and Escherichia coli (ATCC25922), wherein the generation number of the above strains is 4, and PBS containing 0.03mol/L is used for preparing bacterial suspension. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test was carried out according to the neutralization agent identification test 2.1.1.5 in the technical Specification for Disinfection (2002 edition), the test strain was Escherichia coli, and the test was repeated 3 times.

2.3 killing test: the test was carried out according to the test method 2.1.1.7 "Disinfection technical Specification" (2002 edition), using the sample stock solution, the action time was 2.5 minutes, 5 minutes, 7.5 minutes, and the test was repeated 3 times, the test temperature was 21 ℃ to 22 ℃.

3. And (3) testing results:

3.1 coli neutralizer identification result:

in 3 repeated experiments, the original solution is acted for 0.5 minute, and the colony error rates among 3 groups, 4 groups and 5 groups are respectively 4.94 percent, 6.87 percent and 3.83 percent.

Note: negative controls were all grown aseptically.

3.2 results of quantitative bacteria killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of bactericidal component in the disinfectant on Escherichia coli, and the neutralizing agent and the neutralized product have no influence on Escherichia coli and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at 21-22 ℃, the killing logarithm values of escherichia coli, staphylococcus aureus and pseudomonas aeruginosa in suspension are all greater than 5.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Quantitative fungus killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; candida albicans (ATCC10231) with a strain passage number of 4 and a suspension prepared with 0.03mol/L PBS. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test is carried out according to the identification test of a neutralizer 2.1.1.5 in the technical Specification for disinfection (2002 edition), the test strain is candida albicans, the water bath control action temperature is 19-21 ℃, and the test is repeated for 3 times.

2.3 quantitative kill test: according to the operating procedure of a disinfection technical specification (2002 edition) 2.1.1.9 fungus killing test, the sample stock solution is used, the action time is 2.5 minutes, 5 minutes and 7.5 minutes, the action temperature of a water bath is controlled to be 19-21 ℃, and the test is repeated for 3 times.

3. And (3) testing results:

3.1 Candida albicans neutralization agent identification test results:

in 3 repeated experiments, the original solution is acted for 0.5 minute, and the colony error rates among 3 groups, 4 groups and 5 groups are respectively 2.61 percent, 3.04 percent and 2.82 percent.

Note: negative controls were all grown aseptically.

3.2 results of quantitative fungus killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of the bactericidal component in the disinfectant on Candida albicans, and the neutralizing agent and the neutralized product have no influence on Candida albicans and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at the temperature of 19-21 ℃, the killing logarithm value of candida albicans in suspension is greater than 4.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Example 2

Graphene oxide	0.08
		Polyvinylpyrrolidone	1.6
Polyhexamethylene biguanide hydrochloride	0.05
		Purified water	96.4

Adding polyvinylpyrrolidone into a container filled with polyhexamethylene biguanide hydrochloride aqueous solution, stirring to disperse and dissolve, adding graphene oxide, performing ultrasonic treatment to disperse uniformly, and subpackaging in a proper container to obtain the care solution.

And (3) carrying out effect test on the prepared care solution:

quantitative bacteria killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; staphylococcus aureus (ATTCC6538), Pseudomonas aeruginosa (ATCC15442) and Escherichia coli (ATCC25922), wherein the generation number of the above strains is 4, and PBS containing 0.03mol/L is used for preparing bacterial suspension. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test was carried out according to the neutralization agent identification test 2.1.1.5 in the technical Specification for Disinfection (2002 edition), the test strain was Escherichia coli, and the test was repeated 3 times.

2.3 killing test: the test was carried out according to the test method 2.1.1.7 "Disinfection technical Specification" (2002 edition), using the sample stock solution, the action time was 2.5 minutes, 5 minutes, 7.5 minutes, and the test was repeated 3 times, the test temperature was 21 ℃ to 22 ℃.

3. And (3) testing results:

3.1 coli neutralizer identification result:

in 3 repeated experiments, the original solution is acted for 0.5 minute, and the colony error rates among 3 groups, 4 groups and 5 groups are respectively 4.91 percent, 6.85 percent and 3.80 percent.

Note: negative controls were all grown aseptically.

3.2 results of quantitative bacteria killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of bactericidal component in the disinfectant on Escherichia coli, and the neutralizing agent and the neutralized product have no influence on Escherichia coli and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at 21-22 ℃, the killing logarithm values of escherichia coli, staphylococcus aureus and pseudomonas aeruginosa in suspension are all greater than 5.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Quantitative fungus killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; candida albicans (ATCC10231) with a strain passage number of 4 and a suspension prepared with 0.03mol/L PBS. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test is carried out according to the identification test of a neutralizer 2.1.1.5 in the technical Specification for disinfection (2002 edition), the test strain is candida albicans, the water bath control action temperature is 19-21 ℃, and the test is repeated for 3 times.

2.3 quantitative kill test: according to the operating procedure of a disinfection technical specification (2002 edition) 2.1.1.9 fungus killing test, the sample stock solution is used, the action time is 2.5 minutes, 5 minutes and 7.5 minutes, the action temperature of a water bath is controlled to be 19-21 ℃, and the test is repeated for 3 times.

3. And (3) testing results:

3.1 Candida albicans neutralization agent identification test results:

in 3 replicates, the stock solution was applied for 0.5 min, and the colony error rates among 3, 4, and 5 groups were 2.61%, 2.84%, and 2.82%, respectively.

Note: negative controls were all grown aseptically.

3.2 results of quantitative fungus killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of the bactericidal component in the disinfectant on Candida albicans, and the neutralizing agent and the neutralized product have no influence on Candida albicans and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at the temperature of 19-21 ℃, the killing logarithm value of candida albicans in suspension is greater than 4.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Example 3

Graphene oxide	0.065
		Polyvinylpyrrolidone	1
Polyhexamethylene biguanide hydrochloride	0.075
		Purified water	97.75

Adding polyvinylpyrrolidone into a container filled with polyhexamethylene biguanide hydrochloride aqueous solution, stirring to disperse and dissolve, adding graphene oxide, performing ultrasonic treatment to disperse uniformly, and subpackaging in a proper container to obtain the care solution.

And (3) carrying out effect test on the prepared care solution:

quantitative bacteria killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; staphylococcus aureus (ATTCC6538), Pseudomonas aeruginosa (ATCC15442) and Escherichia coli (ATCC25922), wherein the generation number of the above strains is 4, and PBS containing 0.03mol/L is used for preparing bacterial suspension. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test was carried out according to the neutralization agent identification test 2.1.1.5 in the technical Specification for Disinfection (2002 edition), the test strain was Escherichia coli, and the test was repeated 3 times.

2.3 killing test: the test was carried out according to the test method 2.1.1.7 "Disinfection technical Specification" (2002 edition), using the sample stock solution, the action time was 2.5 minutes, 5 minutes, 7.5 minutes, and the test was repeated 3 times, the test temperature was 21 ℃ to 23 ℃.

3. And (3) testing results:

3.1 coli neutralizer identification result:

in 3 replicates, the stock solution was applied for 0.5 min, and the colony error rates among 3, 4, and 5 groups were 4.84%, 6.65%, and 3.29%, respectively.

Note: negative controls were all grown aseptically.

3.2 results of quantitative bacteria killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of bactericidal component in the disinfectant on Escherichia coli, and the neutralizing agent and the neutralized product have no influence on Escherichia coli and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at 21-22 ℃, the killing logarithm values of escherichia coli, staphylococcus aureus and pseudomonas aeruginosa in suspension are all greater than 5.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Quantitative fungus killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; candida albicans (ATCC10231) with a strain passage number of 4 and a suspension prepared with 0.03mol/L PBS. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test is carried out according to the identification test of a neutralizer 2.1.1.5 in the technical Specification for disinfection (2002 edition), the test strain is candida albicans, the water bath control action temperature is 19-21 ℃, and the test is repeated for 3 times.

2.3 quantitative kill test: according to the operating procedure of a disinfection technical specification (2002 edition) 2.1.1.9 fungus killing test, the sample stock solution is used, the action time is 2.5 minutes, 5 minutes and 7.5 minutes, the action temperature of a water bath is controlled to be 19-21 ℃, and the test is repeated for 3 times.

3. And (3) testing results:

3.1 Candida albicans neutralization agent identification test results:

in 3 replicates, the stock solution was applied for 0.5 min, and the colony error rates among 3, 4, and 5 groups were 2.44%, 2.26%, and 2.48%, respectively.

Note: negative controls were all grown aseptically.

3.2 results of quantitative fungus killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of the bactericidal component in the disinfectant on Candida albicans, and the neutralizing agent and the neutralized product have no influence on Candida albicans and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at the temperature of 19-21 ℃, the killing logarithm value of candida albicans in suspension is greater than 4.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Example 4

Nano graphene	0.05
		Polyvinylpyrrolidone	1.6
Polyhexamethylene biguanide hydrochloride	0.1
		Purified water	96.4

Adding polyvinylpyrrolidone into a container filled with polyhexamethylene biguanide hydrochloride aqueous solution, stirring to disperse and dissolve, adding nano graphene, performing ultrasonic treatment to disperse uniformly, and subpackaging in a proper container to obtain the care solution.

And (3) carrying out effect test on the prepared care solution:

quantitative bacteria killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; staphylococcus aureus (ATTCC6538), Pseudomonas aeruginosa (ATCC15442) and Escherichia coli (ATCC25922), wherein the generation number of the above strains is 4, and PBS containing 0.03mol/L is used for preparing bacterial suspension. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test was carried out according to the neutralization agent identification test 2.1.1.5 in the technical Specification for Disinfection (2002 edition), the test strain was Escherichia coli, and the test was repeated 3 times.

2.3 killing test: the test was carried out according to the test method 2.1.1.7 "Disinfection technical Specification" (2002 edition), using the sample stock solution, the action time was 2.5 minutes, 5 minutes, 7.5 minutes, and the test was repeated 3 times, the test temperature was 21 ℃ to 23 ℃.

3. And (3) testing results:

3.1 coli neutralizer identification result:

in 3 replicates, the stock solution was applied for 0.5 min, and the colony error rates between groups 3, 4, and 5 were 4.84%, 6.77%, and 3.73%, respectively.

Note: negative controls were all grown aseptically.

3.2 results of quantitative bacteria killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of bactericidal component in the disinfectant on Escherichia coli, and the neutralizing agent and the neutralized product have no influence on Escherichia coli and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at 21-22 ℃, the killing logarithm values of escherichia coli, staphylococcus aureus and pseudomonas aeruginosa in suspension are all greater than 5.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Quantitative fungus killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; candida albicans (ATCC10231) with a strain passage number of 4 and a suspension prepared with 0.03mol/L PBS. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test is carried out according to the identification test of a neutralizer 2.1.1.5 in the technical Specification for disinfection (2002 edition), the test strain is candida albicans, the water bath control action temperature is 19-21 ℃, and the test is repeated for 3 times.

2.3 quantitative kill test: according to the operating procedure of a disinfection technical specification (2002 edition) 2.1.1.9 fungus killing test, the sample stock solution is used, the action time is 2.5 minutes, 5 minutes and 7.5 minutes, the action temperature of a water bath is controlled to be 19-21 ℃, and the test is repeated for 3 times.

3. And (3) testing results:

3.1 Candida albicans neutralization agent identification test results:

in 3 repeated experiments, the original solution is acted for 0.5 minute, and the colony error rates among 3 groups, 4 groups and 5 groups are respectively 2.31 percent, 2.04 percent and 2.82 percent.

Note: negative controls were all grown aseptically.

3.2 results of quantitative fungus killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of the bactericidal component in the disinfectant on Candida albicans, and the neutralizing agent and the neutralized product have no influence on Candida albicans and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at the temperature of 19-21 ℃, the killing logarithm value of candida albicans in suspension is greater than 4.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Example 5

Nano graphene	0.08
		Polyvinylpyrrolidone	0.5
Polyhexamethylene biguanide hydrochloride	0.05
		Purified water	99.0

Adding polyvinylpyrrolidone into a container filled with polyhexamethylene biguanide hydrochloride aqueous solution, stirring to disperse and dissolve, adding nano graphene, performing ultrasonic treatment to disperse uniformly, and subpackaging in a proper container to obtain the care solution.

And (3) carrying out effect test on the prepared care solution:

quantitative bacteria killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; staphylococcus aureus (ATTCC6538), Pseudomonas aeruginosa (ATCC15442) and Escherichia coli (ATCC25922), wherein the generation number of the above strains is 4, and PBS containing 0.03mol/L is used for preparing bacterial suspension. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test was carried out according to the neutralization agent identification test 2.1.1.5 in the technical Specification for Disinfection (2002 edition), the test strain was Escherichia coli, and the test was repeated 3 times.

2.3 killing test: the test was carried out according to the test method 2.1.1.7 "Disinfection technical Specification" (2002 edition), using the sample stock solution, the action time was 2.5 minutes, 5 minutes, 7.5 minutes, and the test was repeated 3 times, the test temperature was 21 ℃ to 23 ℃.

3. And (3) testing results:

3.1 coli neutralizer identification result:

in 3 repeated experiments, the original solution is acted for 0.5 minute, and the colony error rates among 3 groups, 4 groups and 5 groups are respectively 4.94 percent, 6.87 percent and 3.83 percent.

Note: negative controls were all grown aseptically.

3.2 results of quantitative bacteria killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of bactericidal component in the disinfectant on Escherichia coli, and the neutralizing agent and the neutralized product have no influence on Escherichia coli and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at 21-22 ℃, the killing logarithm values of escherichia coli, staphylococcus aureus and pseudomonas aeruginosa in suspension are all greater than 5.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Quantitative fungus killing test:

1. instruments and strains used for testing:

nutrient agar, a culture dish, a vertical pressure steam sterilizer, an electric heating blowing drying box, a biochemical incubator, an electric heating constant temperature incubator, a scale suction pipe (0.1ml, 1.0ml, 5.0ml, 10.0ml) and the like; candida albicans (ATCC10231) with a strain passage number of 4 and a suspension prepared with 0.03mol/L PBS. Neutralizing agent: D/E neutralizes the broth.

2. The test method comprises the following steps:

2.1 preparation method of bacterial suspension: disinfection Specification (2002 edition) 2.1.1.2.3.

2.2 neutralizer identification test: the test is carried out according to the identification test of a neutralizer 2.1.1.5 in the technical Specification for disinfection (2002 edition), the test strain is candida albicans, the water bath control action temperature is 19-21 ℃, and the test is repeated for 3 times.

2.3 quantitative kill test: according to the operating procedure of a disinfection technical specification (2002 edition) 2.1.1.9 fungus killing test, the sample stock solution is used, the action time is 2.5 minutes, 5 minutes and 7.5 minutes, the action temperature of a water bath is controlled to be 19-21 ℃, and the test is repeated for 3 times.

3. And (3) testing results:

3.1 Candida albicans neutralization agent identification test results:

in 3 replicates, the stock solution was applied for 0.5 min, and the colony error rates among 3, 4, and 5 groups were 2.21%, 2.34%, and 2.72%, respectively.

Note: negative controls were all grown aseptically.

3.2 results of quantitative fungus killing test:

note: 1) negative controls were all grown aseptically.

2)

4. And (4) conclusion:

1) the neutralizing agent (D/E broth) can neutralize the action of the bactericidal component in the disinfectant on Candida albicans, and the neutralizing agent and the neutralized product have no influence on Candida albicans and culture medium.

2) Repeated experiments for 3 times show that the stock solution of the sample acts for 2.5 minutes at the temperature of 19-21 ℃, the killing logarithm value of candida albicans in suspension is greater than 4.00, and the requirements of disinfection technical specifications (2002 edition) are met.

Hereinbefore, specific embodiments of the present application are described with reference to the drawings. However, those skilled in the art will appreciate that various modifications and substitutions can be made to the specific embodiments of the present application without departing from the spirit and scope of the application. Such modifications and substitutions are intended to be within the scope of the present application.