A disinfectant product based on citric acid and natural naphthoquinone compounds

Abstract

The present invention discloses a disinfectant product based on citric acid and natural naphthoquinone compounds and a preparation method thereof. The disinfectant is composed of 15-25% citric acid, 0.01-0.1% natural naphthoquinone compounds, a certain amount of ethanol and a certain amount of deionized water containing alkali metal and alkaline earth metal salts in weight percentage. When in use, the disinfectant can be sprayed on the surface of an article or the diluted disinfectant can be used to clean the article to be disinfected. The disinfectant product has the characteristics of high disinfection and sterilization efficacy, low irritation, low corrosiveness, no odor, and no environmental toxicity of the disinfection residue. The disinfectant is suitable for cold chain disinfection and surface disinfection of vegetables and food. Its application places include public places such as hospitals at all levels, shopping malls, cinemas, Internet cafes, restaurants, administrative offices, enterprises, public transportation, etc., and can also be used for disinfection of homes.

Description

Technical Field

[0001] The present invention specifically relates to a disinfectant product based on citric acid and natural naphthoquinone compounds and a preparation method thereof, and in particular to an environmentally friendly disinfectant liquid and disinfectant gel based on citric acid and natural naphthoquinone and a preparation method thereof. Background Art

[0002] Currently widely used chemical disinfectants such as chlorine-containing disinfectants (84 disinfectant), peracetic acid, ethanol, quaternary ammonium disinfectants, etc. have certain oxidizing, corrosive and allergenic properties.

[0003] Therefore, we urgently need to develop a low-toxic but highly effective disinfectant that is suitable for environmental disinfection protection, while effectively controlling the negative effects of disinfectants on the human body and the environment, and dealing with the increasing number of multidrug-resistant bacteria.

[0004] Natural products are secondary metabolites of plants, animals, and microorganisms. Some natural products possess significant antibacterial and antiviral activity. Natural products are also known for their low toxicity and minimal environmental impact. The use of mugwort for insect repellent and antibacterial purposes has been documented in my country since ancient times. Summary of the Invention

[0005] The present invention aims to overcome the deficiencies of the prior art by providing a disinfectant product based on citric acid and natural naphthoquinone compounds and a method for preparing the same. The disinfectant product is an environmentally friendly, green, low-irritant, and low-corrosive disinfectant or disinfectant gel that can be used for sterilization and disinfection.

[0006] The purpose of the present invention can be achieved by the following solutions:

[0007] The present invention provides a disinfectant product based on citric acid and natural naphthoquinone compounds, the disinfectant product comprising the following components in parts by weight:

[0008]

[0009] As an embodiment of the present invention, the naphthoquinone compound includes one or more of juglone, 7-methyljuglone, 7-methyl-5-acetyljuglone, emodin, and shikonin.

[0010] As one embodiment of the present invention, the citric acid has a purity greater than 99.5%.

[0011] As an embodiment of the present invention, the disinfection product further comprises 0.1 to 1 parts of inorganic salts by weight.

[0012] In one embodiment of the present invention, the inorganic salt comprises an alkali metal salt and / or an alkaline earth metal salt. The inorganic salt is analytically pure. Alkali metal and alkaline earth metal salts can absorb moisture from the air through deliquescent effects, thereby extending the effective duration of the disinfectant during daily use and enhancing its long-term antibacterial ability.

[0013] As an embodiment of the present invention, the alkali metal salt includes one or more of sodium chloride, potassium sulfate, and potassium nitrate; the alkaline earth metal salt includes one or more of magnesium chloride, magnesium sulfate, and calcium sulfate.

[0014] As an embodiment of the present invention, the disinfection product further comprises 1 to 5 parts by weight of a cosolvent, wherein the cosolvent comprises one of ethanol, isopropanol, and DMSO.

[0015] The present invention also provides an application of a disinfectant product based on citric acid and natural naphthoquinone compounds in the preparation of disinfectant liquid and disinfectant gel.

[0016] The present invention provides a disinfectant, which comprises the following components in parts by weight per hundred parts:

[0017] 15-25 parts of citric acid;

[0018] 1-5 parts of cosolvent;

[0019] 0.01~1 part of naphthoquinone compound;

[0020] 0.1~1 part of inorganic salt;

[0021] Deionized water balance.

[0022] In one embodiment of the present invention, the cosolvent comprises one of ethanol, isopropanol, and DMSO. The naphthoquinone compound is dissolved in the cosolvent and then mixed with other components of the disinfectant to achieve a mass fraction of the naphthoquinone compound in the disinfectant of 0.01 to 1%.

[0023] The present invention also provides a method for preparing the disinfectant, which comprises the steps of:

[0024] S1. Weigh citric acid, add it to an appropriate amount of deionized water, and heat it to 70-95°C in a water bath. Ensure that all the citric acid is dissolved during the preparation process.

[0025] S2. Weigh an inorganic salt, add it to the citric acid aqueous solution prepared in step S1, and stir at 70-95° C. for 25-35 minutes at a stirring speed of 60-100 r / min.

[0026] S3. Cool the solution obtained in step S2 to room temperature, add a mixed solution formed by dissolving the naphthoquinone compound in a cosolvent, so that the mass fraction of the naphthoquinone compound in the disinfectant is 0.01-1%, stir evenly, and the resulting solution is the disinfectant.

[0027] The present invention provides a disinfectant gel, wherein the disinfectant comprises the following components in parts by weight per hundred parts:

[0028] 15-25 parts of citric acid;

[0029] 0.01~0.1 parts of naphthoquinone compounds;

[0030] 0.1~1 part of inorganic salt;

[0031] 1-5 parts of cosolvent;

[0032] 1~3 parts thickener;

[0033] Deionized water balance.

[0034] In one embodiment of the present invention, the cosolvent comprises one of ethanol, isopropanol, and DMSO. The naphthoquinone compound is dissolved in the cosolvent and then mixed with other components of the disinfectant to achieve a mass fraction of the naphthoquinone compound in the disinfectant of 0.01 to 1%.

[0035] In one embodiment of the present invention, the thickener is formed by adding hydroxyethyl cellulose to an ethanol solution containing deionized water, acetate buffer, and chlorhexidine acetate. The ratio of hydroxyethyl cellulose, deionized water, acetate buffer, and chlorhexidine acetate is 10g:18.5ml:1ml:0.5ml. The addition of the thickening component imparts a gel-like consistency to the disinfectant, making it easier to carry and use.

[0036] The present invention also provides a method for preparing a disinfectant gel, the method comprising the steps of:

[0037] A1. Weigh citric acid and add it to an appropriate amount of deionized water. Heat the mixture in a water bath to 70-95°C, ensuring that all the citric acid is dissolved.

[0038] A2. Weigh an inorganic salt and add it to the citric acid aqueous solution obtained in step A1. Stir at 70-95°C for 25-35 minutes at a stirring speed of 60-100 r / min.

[0039] A3. Cool the solution obtained in step A2 to room temperature, add a mixed solution formed by dissolving the naphthoquinone compound in a cosolvent to adjust the mass fraction of the naphthoquinone compound in the disinfectant to 0.01-1%, stir evenly, add a thickener, mix and thicken to obtain a gel, and obtain a disinfectant gel.

[0040] The preparation method of the present invention can significantly improve the solubility of salts.

[0041] The preparation method described in the present invention is conducive to enhancing the synergistic effect of citric acid and inorganic salts, mainly because in step A2, citric acid and inorganic salts can undergo a complex reaction under heating and stirring, which helps to increase the concentration of effective bactericidal ingredients in the disinfectant. The dosage of each component in the present invention is the optimal dosage. The dosage of citric acid is at its maximum solubility. Excessive addition of citric acid will form crystals at room temperature, reducing the effect; too little dosage will reduce its concentration and reduce the bactericidal effect. The dosage of naphthoquinone compounds is also at its maximum solubility. If added too much, substances in the disinfectant will precipitate and reduce the stability of the disinfectant. Too little dosage will reduce its concentration and reduce the bactericidal effect. Excessive dosage of inorganic salts will lead to reduced stability of the disinfectant, while too little dosage is not conducive to prolonging the bactericidal effect of the disinfectant.

[0042] The disinfectant solution prepared by the invention can be sprayed on the surface of objects and has the characteristics of disinfection and sterilization, low irritation, low corrosiveness, no odor, and no environmental toxicity of disinfection residues.

[0043] Compared with the prior art, the present invention has the following beneficial effects:

[0044] 1) Citric acid, a natural plant ingredient, has the disinfection effect of killing viruses and bacteria, and contains a certain acidity, is green and low-toxic.

[0045] 2) By adding natural naphthoquinone compounds, new antibacterial and antiviral targets are provided, enhancing the efficiency of disinfectants in killing bacteria and viruses.

[0046] 3) A new green, environmentally friendly, safe and efficient disinfectant is prepared by using low-toxic and pollution-free citric acid, natural naphthoquinone compounds and inorganic salts.

[0047] 4) Through the pore-punching effect of citric acid, natural naphthoquinone compounds can freely diffuse into bacterial cells, thereby improving the bactericidal efficiency of naphthoquinone compounds.

[0048] 5) Citric acid and naphthoquinone compounds have good synergistic effects and are low in toxicity, making them the preferred formulation for disinfectants. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Other features, objects and advantages of the present invention will become more apparent upon reading the detailed description of non-limiting embodiments with reference to the following drawings:

[0050] Figure 1 Schematic diagram of the minimum inhibitory concentration of the disinfectants prepared in Example 1 and Comparative Examples 1-4;

[0051] Figure 2Schematic diagram of the antibacterial time of Example 1 and Comparative Example 5;

[0052] Figure 3 This is a schematic diagram of the antibacterial effect of disinfectant in Example 1;

[0053] Figure 4 This is a schematic diagram of the disinfection effect of Example 1;

[0054] Figure 5 This is a graph showing the changes in zebrafish heart rate after treatment with different concentrations of disinfectant in Example 1;

[0055] Figure 6 This is a graph showing the change in the number of tail flicks of zebrafish after treatment with different concentrations of disinfectant in Example 1;

[0056] Figure 7 This is a graph showing the weight changes of mice after being fed with different concentrations of disinfectant in Example 1. DETAILED DESCRIPTION

[0057] The present invention is described in detail below in conjunction with the accompanying drawings and specific examples. The following examples will help those skilled in the art further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that those skilled in the art can make several adjustments and improvements without departing from the scope of the present invention. These all fall within the scope of protection of the present invention. The experimental methods in the following examples where specific conditions are not specified are generally based on conventional conditions or the conditions recommended by the manufacturer.

[0058] The present invention discloses a disinfectant based on citric acid and natural naphthoquinone and a method for preparing the same. Those skilled in the art may refer to the present invention and appropriately modify the process parameters. It is particularly important to note that all similar substitutions and modifications obvious to those skilled in the art are considered to be included in the present invention.

[0059] Example 1

[0060] This embodiment relates to a method for preparing a disinfectant based on citric acid and natural naphthoquinone compounds, and the preparation steps are as follows:

[0061] (1) Accurately weigh 48 g of citric acid (purity greater than 99.5%), 0.7 g of analytical grade NaCl, 2 mL of 100 μM juglone ethanol solution, and 200 mL of purified water, and set aside.

[0062] (2) Add citric acid and NaCl to 200 mL of purified water and heat to 70-95°C in a water bath reactor. Ensure that all citric acid is dissolved during the preparation process.

[0063] (3) Cool the solution obtained in step (2) to room temperature, add 2 mL of 100 μM juglone ethanol solution, and stir evenly. The resulting solution is a disinfectant based on citric acid and natural naphthoquinone;

[0064] (4) The final solution is divided into spray bottles.

[0065] Example 2

[0066] This embodiment relates to a method for preparing a disinfectant gel based on citric acid and natural naphthoquinone compounds.

[0067] (1) Accurately weigh 40 g of citric acid with a purity greater than 99.5%, 0.3 g of analytical grade potassium sulfate, 2 mL of 100 μM emodin ethanol solution, and 200 mL of purified water, and set aside;

[0068] (2) Add citric acid and potassium sulfate to 200 mL of purified water and heat to 70-95°C in a water bath reactor. Ensure that all citric acid is dissolved during the preparation process.

[0069] (3) Cool the solution obtained in step (2) to room temperature, add 2 mL of 100 μM emodin ethanol solution, and stir evenly;

[0070] (4) Add 3% by mass of a thickener to the solution obtained in step (3), wherein the ratio of hydroxyethyl cellulose, deionized water, acetate buffer, and acetic acid chloride in the thickener is 10 g:18.5 ml:1 ml:0.5 ml, and thicken to obtain a gel, i.e., a disinfectant gel;

[0071] (5) The final gel is divided into bottles.

[0072] Example 3

[0073] This embodiment relates to a method for preparing a disinfectant based on citric acid and natural naphthoquinone compounds.

[0074] (1) Accurately weigh 38 g of citric acid (purity greater than 99.5%), 0.28 g of analytical grade NaCl, 2 mL of 100 μM 7-methyljuglone ethanol solution, and 198 mL of purified water, and set aside.

[0075] (2) Add citric acid and NaCl to 198 mL of purified water and heat to 70-95 °C in a water bath reactor. Ensure that all citric acid is dissolved during the preparation process.

[0076] (3) Cool the solution obtained in step (2) to room temperature, add 2 mL of 100 μM 7-methyljuglone ethanol solution, and stir evenly. The resulting solution is a disinfectant based on citric acid and natural naphthoquinone;

[0077] (4) The final solution is divided into spray bottles.

[0078] Example 4

[0079] This embodiment relates to a method for preparing a disinfectant based on citric acid and natural naphthoquinone compounds.

[0080] (1) Accurately weigh 64 g of citric acid (purity greater than 99.5%), 0.28 g of analytical grade NaCl, 0.1 mL of 100 μM 7-methyl-5-acetyljuglans quinone ethanol solution, and 200 mL of purified water, and set aside.

[0081] (2) Add citric acid and NaCl to 200 mL of purified water and heat to 70-95°C in a water bath reactor. Ensure that all citric acid is dissolved during the preparation process.

[0082] (3) Cool the solution obtained in step (2) to room temperature, add 0.1 mL of 100 μM 7-methyl-5-acetyljuglans quinone ethanol solution, and stir evenly. The resulting solution is a disinfectant based on citric acid and natural naphthoquinone;

[0083] (4) The final solution is divided into spray bottles.

[0084] Example 5

[0085] This embodiment relates to a method for preparing a disinfectant based on citric acid and natural naphthoquinone compounds.

[0086] (1) Accurately weigh 38 g of citric acid (purity greater than 99.5%), 0.54 g of analytical grade NaCl, 2 mL of 100 μM shikonin ethanol solution, and 198 mL of purified water, and set aside.

[0087] (2) Add citric acid and NaCl to 198 mL of purified water and heat to 70-95 °C in a water bath reactor. Ensure that all citric acid is dissolved during the preparation process.

[0088] (3) Cool the solution obtained in step (2) to room temperature, add 2 mL of 100 μM shikonin ethanol solution, and stir evenly. The resulting solution is a disinfectant based on citric acid and natural naphthoquinone;

[0089] (4) The final solution is divided into spray bottles.

[0090] Comparative Example 1

[0091] This comparative example relates to a method for preparing a disinfectant based on citric acid and natural naphthoquinone compounds. The method is substantially the same as that in Example 1, except that 20 g of citric acid and 198 mL of purified water are used.

[0092] The MIC (minimum inhibitory concentration) of the disinfectant was determined by the same 96-well plate method as in Example 1. The MIC of the composite disinfectant in Comparative Example 1 was 100-fold diluted, and the disinfection effect was reduced compared with the original disinfectant.

[0093] Comparative Example 2

[0094] This comparative example relates to a method for preparing a disinfectant based on citric acid and natural naphthoquinone compounds. The method is substantially the same as that of Example 1, except that juglone is not contained.

[0095] The MIC (minimum inhibitory concentration) of the disinfectant was determined by the same 96-well plate method as in Example 1. The MIC of the composite disinfectant in Comparative Example 2 was 150 times diluted, and the disinfection effect was reduced compared with the original disinfectant.

[0096] Comparative Example 3

[0097] This comparative example relates to a method for preparing a disinfectant based on citric acid and natural naphthoquinone compounds. The method is substantially the same as that of Example 1, except that juglone is replaced with 1,4-naphthoquinone.

[0098] The MIC (minimum inhibitory concentration) of the disinfectant was determined by the same 96-well plate method as in Example 1. The MIC of the composite disinfectant in Comparative Example 3 was 100 times diluted, and the disinfection effect was reduced compared with the original disinfectant. In addition, juglone was more economical.

[0099] Comparative Example 4

[0100] This comparative example relates to a method for preparing a disinfectant based on citric acid and natural naphthoquinone compounds. The method is substantially the same as that of Example 1, except that citric acid is replaced by lactic acid.

[0101] The MIC (minimum inhibitory concentration) of the disinfectant was determined by the same 96-well plate method as in Example 1. The MIC of the composite disinfectant in Comparative Example 4 was 10-fold diluted, and the disinfection effect was reduced compared with the original disinfectant.

[0102] like Figure 1 As shown, the disinfection and antibacterial effects of Comparative Examples 1-4 were significantly reduced compared to Example 1. The bactericidal effect of organic acids such as lactic acid was not as good as that of citric acid, and the complexation effect of other acids with salts was poor.

[0103] Comparative Example 5

[0104] This comparative example relates to a method for preparing a disinfectant based on citric acid and natural naphthoquinone compounds. The method is substantially the same as that of Example 1, except that no inorganic salt is added.

[0105] The MIC (minimum inhibitory concentration) of the disinfectant was determined by the same 96-well plate method as in Example 1. The MIC of the composite disinfectant in Comparative Example 5 was 10 times diluted, and the disinfection effect was reduced compared with the original disinfectant. Figure 2 As shown in Figure 3, the effective antibacterial time was significantly shortened.

[0106] Effect verification 1: Antibacterial kinetics experiment of the disinfectant prepared in Example 1.

[0107] This experiment relates to the antibacterial kinetics experimental methods and results of the disinfectant based on citric acid and natural naphthoquinone compounds in Example 1. It is characterized by resuspending the bacteria in LB medium and diluting the disinfectant in LB medium to provide sufficient growth conditions for the bacteria, thereby simulating the inhibition of bacterial growth by disinfectant in a nutrient-rich environment.

[0108] According to the American CLSI Association standards, the antibacterial effect of the compound disinfectant was evaluated by the improved microplate method. The specific experimental steps are as follows:

[0109] (1) Take a suspension of Staphylococcus aureus in the logarithmic growth phase that has been cultured overnight and use a microplate reader to measure its OD600 value. If its OD value is between 0.5 and 0.8, it is considered to be in the logarithmic growth phase.

[0110] (2) Dilute the bacterial suspension 100-fold using high-temperature sterilized LB medium under sterile conditions, and then use a dispenser to add 100uL of the diluted bacterial suspension to each microwell in the 96-well plate.

[0111] (3) Add 100 μL of 50-fold diluted disinfectant water of the high-temperature sterilized LB medium into each of the eighteen parallel bacterial suspension microwells.

[0112] (4) Place the 96-well plate in a constant temperature incubator at 37°C and culture. Aspirate the bacterial suspensions in the three wells at 0h, 1h, 2h, 4h, and 8h, respectively. Then use a pipette to transfer the bacterial suspensions in the microwells corresponding to different concentration gradients into different 1.5mL PE tubes, and immediately place the PE tubes in ice.

[0113] (5) Centrifuge the bacterial suspension at 10,000 rpm for 10 min using a high-speed centrifuge and resuspend in 0.85% NaCl solution. Repeat this process three times.

[0114] (6) Stain the bacteria using LIVE\DEAD stain in the dark and incubate at room temperature for 15 min.

[0115] (7) Flow cytometry was used to determine the bacterial population and count the bacteria. The antibacterial activity of the compound disinfectant was characterized by the number of live and dead cells under different concentration gradients. Figure 3 shown.

[0116] The experimental results show that the removal rate of Staphylococcus aureus increases with increasing treatment time. While the removal rate was less than 40% at one hour, it exceeded 90% after two hours of treatment. The inhibition rate continued to rise over the next eight hours, reaching 98.3% at that time. This result demonstrates that the new disinfectant has the ability to stably and continuously inhibit bacterial growth.

[0117] Effect verification 2: bactericidal kinetics experiment of the disinfectant prepared in Example 1.

[0118] This experiment relates to the experimental methods and results for the bactericidal kinetics of a disinfectant based on citric acid and natural naphthoquinone compounds, described in Example 1. During the assay, the bacteria were resuspended in a 0.85% NaCl solution, and the disinfectant was also diluted with a 0.85% NaCl solution, depriving the bacteria of the organic matter and other energy sources they need to survive and thus preventing bacterial growth. Under these conditions, the disinfectant's direct bacterial killing effect was measured in a nutrient-deprived environment.

[0119] According to the American CLSI Association standards, the disinfection effect of the compound disinfectant was evaluated by the improved microplate method. The specific experimental steps are as follows:

[0120] (1) Take a suspension of Staphylococcus aureus in the logarithmic growth phase that has been cultured overnight and use a microplate reader to measure its OD600 value. If its OD value is between 0.5 and 0.8, it is considered to be in the logarithmic growth phase.

[0121] (2) Dilute the bacterial suspension 100-fold using high-temperature sterilized 0.85% NaCl solution under sterile conditions, and then use a dispenser to add 100 μL of the diluted bacterial suspension to each microwell in the 96-well plate.

[0122] (3) Add 100 μL of disinfectant water diluted 50 times with high-temperature sterilized 0.85% NaCl solution into 18 parallel bacterial suspension microwells.

[0123] (4) Place the 96-well plate in a constant temperature incubator at 37°C and culture the bacterial suspensions in the three wells at 0h, 15min, 30min, 1h, 2h, and 4h, respectively. Then use a pipette to transfer the bacterial suspensions in the microwells corresponding to different concentration gradients into different 1.5mL PE tubes, and immediately place the PE tubes in ice.

[0124] (5) Centrifuge the bacterial suspension at 10,000 rpm for 10 min using a high-speed centrifuge and resuspend in 0.85% NaCl solution. Repeat this process three times.

[0125] (6) Stain the bacteria using LIVE\DEAD stain in the dark and incubate at room temperature for 15 min.

[0126] (7) Flow cytometry was used to determine the bacterial population and bacterial technology was performed to characterize the antibacterial activity of the composite disinfectant according to the characteristics of living cells and dead cells under different concentration gradients. The experimental results are as follows: Figure 4 shown.

[0127] From the experimental results, we can observe that the removal rate of Staphylococcus aureus increases with increasing treatment time with the novel disinfectant of the present invention. At 15 minutes, the removal rate of Staphylococcus aureus reached nearly 70%, and at 240 minutes, it reached 96.7%. Furthermore, the bacteria detected by flow cytometry all had severely damaged cell membranes, significantly reducing their viability and pathogenicity. This result demonstrates that the novel disinfectant of the present invention has a relatively strong ability to kill bacteria in a short period of time.

[0128] Effect verification three: skin irritation experiment of the disinfectant prepared in Example 1.

[0129] This experiment involves the skin irritation experiment of the disinfectant based on citric acid and natural naphthoquinone compounds in Example 1.

[0130] This experiment referred to the "Procedures and Methods for Toxicological Evaluation of Safety of Disinfectants" (GB / T 38496-2020) and used New Zealand rabbits to conduct a safety evaluation of disinfectants based on citric acid and natural naphthoquinone compounds.

[0131] The specific steps of the experiment are as follows:

[0132] Take an adult New Zealand rabbit, and use scissors to remove the hair on both sides of the spine of the New Zealand rabbit's back 24 hours before the start of the experiment. Soak 1mL of the test substance on 2~4 layers of gauze with an area of 2.5cm*2.5cm, apply it to the fur surface on one side, and then fix it with non-irritating plastic film and tape. The experimental group has a total of six groups: 20 times diluted disinfectant, 10 times diluted disinfectant, 5 times diluted disinfectant, 20 times diluted 84, 10 times diluted 84, 5 times diluted 84, and a blank control group. After 4 hours of application, the residual test substance was removed with a non-irritating solvent. The local skin reaction was observed 1h, 24h, and 48h after the removal of the test substance, and scored according to the skin irritation score table in the "Procedures and Methods for Safety Toxicological Evaluation of Disinfectants" (GB / T 38496-2020). The specific scoring criteria are as follows:

[0133] The results of a complete skin irritation test are as follows:

[0134] Table 1: Scores of different test substances in a complete skin irritation test

[0135]

[0136] From the experimental result, we can observe that after 20% 84 disinfectants are processed, the formation of erythema and edema is all comparatively obvious during 1 hour, and particularly erythema is comparatively serious, and continues to 24 hours, and the situation of redness and swelling is just slightly weakened after 48 hours. The novel disinfectant of the present invention with the same concentration has only appeared the erythema and edema that are barely visible, and after 48 hours, skin has fully recovered to normal. The New Zealand rabbit skin after 10% 84 disinfectants and 5% 84 disinfectants are processed has all appeared erythema and edema in varying degrees, but erythema and edema are not then appeared in the composite disinfectant of the same concentration. This result proves that the novel disinfectant of the present invention is very little to the skin irritation of rabbits under effective bactericidal concentration, and is far lower than 84 disinfectants, proving that the safety of novel disinfectant of the present invention is higher.

[0137] Effect verification 4: Ecotoxicity evaluation experiment of the disinfectant prepared in Example 1.

[0138] This experiment involves the ecotoxicity evaluation of the disinfectant based on citric acid and natural naphthoquinone compounds in Example 1.

[0139] This experiment uses a zebrafish model to conduct environmental toxicity assessment, including acute toxicity tests and behavioral experiments. The specific experimental steps are as follows:

[0140] 1. Acute toxicity test

[0141] (1) Use OD water to dilute the compound disinfectant and 84 disinfectant to a final concentration of 20%, 10%, 5%, and 1%, respectively (calculate the volume ratio based on the working concentration).

[0142] (2) Twenty zebrafish embryos and 20 zebrafish larvae were added to OD water containing different concentrations of disinfectant, and the survival of the zebrafish embryos and larvae was observed at 5 minutes, 30 minutes, and 90 minutes.

[0143] The results of the zebrafish acute toxicity test are as follows:

[0144] Table 2: Results of acute toxicity test on zebrafish

[0145]

[0146] The results are shown in Table 2. It can be seen that at the same treatment concentration, the lethality of the disinfectant of the present invention was lower than that of the disinfectant 84 for both zebrafish embryos and juveniles. At the experimental concentrations used in the experiment, no embryos treated with the disinfectant of the present invention died, while embryos treated with the disinfectant 84 at concentrations of 20% and 10% died. Furthermore, the median lethal concentration of the disinfectant of the present invention for zebrafish juveniles was between 5% and 10%, while that of the disinfectant 84 for zebrafish juveniles was less than 1%. This result demonstrates that the acute toxicity of the novel disinfectant of the present invention to zebrafish is significantly lower than that of the disinfectant 84.

[0147] 2. Zebrafish behavioral experiments

[0148] (1) Use OD water to dilute the composite disinfectant and 84 disinfectant to a final concentration of 0.5%, 0.25%, 0.125%, 0.6125%, and 0.30625%, respectively (volume ratio calculated based on the working concentration).

[0149] (2) Add 20 zebrafish embryos and 20 zebrafish larvae to water containing different concentrations of disinfectant. The number of tail flicks per minute of the embryos was recorded at 24 hours, and the number of heart beats per minute of the larvae was recorded at 48 hours to assess whether the zebrafish's condition was affected by the disinfectant. All data were recorded in the form of video. (Because zebrafish cannot survive long-term in 0.5% 84 disinfectant water, no 84 positive control group was set up.)

[0150] (3) The video was processed using ImageJ software to obtain the tail swinging data of zebrafish embryos per minute and the heartbeat data of zebrafish larvae.

[0151] Related experimental results such as Figure 5 、 Figure 6As shown: According to the experimental results, it can be seen that there is no significant difference in the basic behavior of the zebrafish in the experimental group treated with the new disinfectant of the present invention and the zebrafish in the blank control group. It can be considered that the experimental concentration has no effect on the growth of zebrafish, which proves that the ecological toxicity of the new disinfectant of the present invention is low.

[0152] Effect Verification 5: In vivo toxicity evaluation experiment of the disinfectant prepared in Example 1.

[0153] This experiment involves the in vivo toxicity of the disinfectant based on citric acid and natural naphthoquinone compounds described in Example 1. The toxicity of the disinfectant in Balb / C mice was tested by feeding the mice with disinfectant water.

[0154] We continuously monitored the weight changes of mice for seven days and recorded the following: Figure 7 shown.

[0155] From the experimental results, we observed that under the feeding conditions of the disinfectant of the present invention at an effective bactericidal concentration, the weight changes of mice within seven days were basically the same as those of mice in the blank control group, proving that the oral toxicity of the new disinfectant of the present invention is low and relatively safe.

[0156] In summary, the disinfectant based on citric acid and natural naphthoquinone compounds described in the present invention has a simple preparation method and readily available raw materials. Activity assay results show that the disinfectant has the characteristics of high disinfection and sterilization efficacy, low irritation, low corrosiveness, no odor, no environmental toxicity of disinfection residues, and stable quality. This disinfectant is suitable for cold chain disinfection and protection of vegetable and food surface disinfection. Its application places include public places such as hospitals at all levels, shopping malls, cinemas, Internet cafes, restaurants, administrative offices, enterprises, public transportation, and can also be used for disinfection of homes.

[0157] The above describes the specific embodiments of the present invention. It should be understood that the present invention is not limited to the above specific embodiments, and those skilled in the art may make various variations or modifications within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. A disinfectant, characterized in that: The disinfectant comprises the following components in parts by weight per hundred parts: 15-25 parts of citric acid; 1-5 parts of cosolvent; 0.01~1 part of naphthoquinone compound; 0.1~1 part of inorganic salt; Deionized water balance; The naphthoquinone compound is juglone; The inorganic salt is sodium chloride; The preparation method of the disinfectant comprises the following steps: S1. Weigh citric acid, add it to an appropriate amount of deionized water, and heat it to 70-95°C in a water bath reactor. Ensure that all the citric acid is dissolved during the preparation process. S2. Weigh the inorganic salt and add it to the citric acid aqueous solution prepared in step S1. Stir at 70-95°C for 25-35 minutes at a stirring speed of 60-100 r / min. S3. Cool the solution obtained in step S2 to room temperature, add a mixed solution formed by dissolving the naphthoquinone compound in a cosolvent, so that the mass fraction of the naphthoquinone compound in the disinfectant is 0.01-1%, stir evenly, and the resulting solution is the disinfectant.

2. A method for preparing the disinfectant according to claim 1, characterized in that: The method comprises the following steps: S1. Weigh citric acid, add it to an appropriate amount of deionized water, and heat it to 70-95°C in a water bath reactor. Ensure that all the citric acid is dissolved during the preparation process. S2. Weigh the inorganic salt and add it to the citric acid aqueous solution prepared in step S1. Stir at 70-95°C for 25-35 minutes at a stirring speed of 60-100 r / min. S3. Cool the solution obtained in step S2 to room temperature, add a mixed solution formed by dissolving the naphthoquinone compound in a cosolvent, so that the mass fraction of the naphthoquinone compound in the disinfectant is 0.01-1%, stir evenly, and the resulting solution is the disinfectant.

3. A disinfectant gel, characterized in that: The disinfectant gel comprises the following components in parts by weight per hundred: 15-25 parts of citric acid; 0.01~0.1 parts of naphthoquinone compounds; 0.1~1 part of inorganic salt; 1-5 parts of cosolvent; 1~3 parts thickener; Deionized water balance; The naphthoquinone compound is juglone; The inorganic salt is sodium chloride; The preparation method of the disinfectant gel comprises the following steps: A1. Weigh citric acid and add it to an appropriate amount of deionized water. Heat the mixture in a water bath to 70-95°C, ensuring that all the citric acid is dissolved. A2. Weigh an inorganic salt and add it to the citric acid aqueous solution obtained in step A1. Stir at 70-95°C for 25-35 minutes at a stirring speed of 60-100 r / min. A3. Cool the solution obtained in step A2 to room temperature, add a mixed solution formed by dissolving the naphthoquinone compound in a cosolvent to adjust the mass fraction of the naphthoquinone compound in the disinfectant to 0.01-1%, stir evenly, add a thickener, mix and thicken to obtain a gel, and obtain a disinfectant gel.

4. A method for preparing the disinfectant gel according to claim 3, characterized in that: The method comprises the following steps: A1. Weigh citric acid and add it to an appropriate amount of deionized water. Heat the mixture in a water bath to 70-95°C, ensuring that all the citric acid is dissolved. A2. Weigh an inorganic salt and add it to the citric acid aqueous solution obtained in step A1. Stir at 70-95°C for 25-35 minutes at a stirring speed of 60-100 r / min. A3. Cool the solution obtained in step A2 to room temperature, add a mixed solution formed by dissolving the naphthoquinone compound in a cosolvent to adjust the mass fraction of the naphthoquinone compound in the disinfectant to 0.01-1%, stir evenly, add a thickener, mix and thicken to obtain a gel, and obtain a disinfectant gel.

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