Composite potassium persulfate disinfection enhancer and its application

Abstract

The present invention belongs to the technical field of disinfection and sterilization, and discloses a composite disinfection composition, including potassium hydrogen sulfate and wintergreen essential oil. When the above two are used separately, the antibacterial effect is weak, and after the combined action, there is an obvious synergistic antibacterial effect. Therefore, the present application further adds the above two to a solvent and dissolves them to obtain a novel potassium persulfate synergist. Compared with potassium persulfate disinfection alone, the present invention can kill bacteria more efficiently and reduce the use of potassium persulfate concentration, further expanding the application of potassium persulfate in disinfection and sterilization products.

Description

Technical Field

[0001] The invention belongs to the technical field of disinfection and sterilization, and more specifically, relates to a novel potassium persulfate enhancer and application thereof. Background Art

[0002] Potassium persulfate is a white crystalline powder. It is a strong oxidant and is widely used in many chemical reactions and experiments. Potassium persulfate is mainly used as an initiator, especially in the preparation of polymers and resins. It is also used in water treatment, luminophore preparation, paint and ink drying processes, and as a disinfectant. In the polymer industry, potassium persulfate is often used as a free radical initiator. Potassium persulfate is used as a water treatment agent that can oxidize and remove organic and inorganic pollutants from water. This is common in water treatment processes in industry and laboratories.

[0003] In addition, potassium persulfate has some inevitable limitations: 1. The bactericidal spectrum of potassium persulfate at low doses is limited, and it can only kill some viruses and bacteria (with envelopes). In order to enhance the bactericidal effect of potassium persulfate, studying synergists is an indispensable step. However, there are still few related synergists reported so far. The development of potassium persulfate disinfectant synergists can further expand the application of potassium persulfate in the field of disinfection. Summary of the invention

[0004] In order to solve the above problems in the prior art, the present invention provides a novel potassium persulfate synergist. Potassium persulfate is one of the most commonly used disinfectants at present. The combination of potassium persulfate and essential oil provided by the present invention can significantly enhance the bactericidal effect of potassium persulfate. The present invention uses Escherichia coli ATCC25922 and Staphylococcus aureus ATCC29213 as representative strains of Gram-negative bacteria and Gram-positive bacteria, and proves that wintergreen essential oil can be used as a bactericidal synergist of potassium persulfate.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] A composite disinfection composition comprises potassium persulfate and wintergreen essential oil.

[0007] Wintergreen essential oil is a plant essence extracted from the leaves or flowers of holly. It has a refreshing and uplifting aroma with a strong minty flavor and is generally light yellow in color. Its main ingredients are methyl salicylate and a small amount of organic ingredients, such as myrcene, limonene 3,7-guaiacdiene, etc. It has anti-inflammatory and analgesic effects, can promote local blood circulation, refresh the mind and calm the spirit, and also has bactericidal, diuretic, anti-rheumatic, and blood circulation promoting effects. Wintergreen essential oil needs to be used in low doses and is prohibited for pregnant women and epileptic patients. It is currently widely used in food, pesticides and cosmetics.

[0008] Essential oils are highly concentrated natural oils extracted from plants, which contain the aromatic components and active ingredients of the plants. They have rich chemical compositions and special aromas, so they are widely used in medicine, health care, cosmetics, spices and food. Essential oils are obtained through various extraction methods, and common extraction methods include distillation, pressing, solvent extraction and cold pressing. Each extraction method will affect the composition and purity of the essential oil. Antibacterial effect is one of the important functions of essential oils. Many essential oils have antibacterial, antifungal and antiviral abilities, which can effectively inhibit the growth and reproduction of microorganisms.

[0009] The present application dissolves the composite disinfection composition in a solvent to obtain the disinfection enhancer. Further, the solvent is water.

[0010] Preferably, the concentration of potassium persulfate is between 0.01% and 1%.

[0011] Preferably, the concentration of wintergreen essential oil is 0.1%-2%. The present invention uses Escherichia coli ATCC25922 and Staphylococcus aureus ATCC29213 as representative strains of Gram-negative bacteria and Gram-positive bacteria to prove that wintergreen essential oil can be used as a bactericidal synergist of potassium persulfate.

[0012] Compared with the prior art, the present invention has the following beneficial effects:

[0013] The present application provides a composite disinfection composition, including potassium persulfate and wintergreen essential oil. When the above two are used separately, the antibacterial effect is weak, and when they are used together, there is an obvious synergistic antibacterial effect. Therefore, the present application further adds the above two to a solvent and dissolves them to obtain a new potassium persulfate synergist. Compared with potassium persulfate disinfection alone, the present invention can kill bacteria more efficiently and reduce the concentration of potassium persulfate, further expanding the application of potassium persulfate in disinfection and sterilization products. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is the bactericidal effect of wintergreen essential oil combined with potassium persulfate on Escherichia coli ATCC25922.

[0015] Figure 2 It is the bactericidal effect of wintergreen essential oil combined with potassium persulfate against Staphylococcus aureus ATCC29213. DETAILED DESCRIPTION

[0016] The technical scheme of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention and the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of the present invention. If the specific conditions are not specified in the embodiments, they are prepared according to the conditions recommended by the normal conditions or the manufacturers. If the manufacturer is not specified in the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0017] Example 1 Determination of MIC of Wintergreen Essential Oil and Potassium Persulfate against ATCC 25922 and ATCC 29213 Strains

[0018] 1. Experimental materials:

[0019] (1) Potassium persulfate, wintergreen essential oil

[0020] Experimental: The autoclaved MH broth was cooled and set aside.

[0021] (2) Resin Azure: Macklin brand, MW 251.17, purity 90%;

[0022] (3) Escherichia coli standard strains ATCC25922 and ATCC29213 (laboratory collection).

[0023] 2. Preparation before the test:

[0024] (1) Take wintergreen essential oil out of the refrigerator and equilibrate it to room temperature, and draw a portion for later use;

[0025] (2) Escherichia coli standard strains ATCC 25922 and ATCC 29213 were cultured on LB agar medium until they reached an appropriate size.

[0026] (3) Weigh 0.2512 g of resin azure powder into a centrifuge tube and add 10 mL of pure water to dissolve it. The concentration of the resin azure solution is 10 mM / L.

[0027] 3. Wintergreen essential oil / potassium persulfate bactericidal effect evaluation experiment:

[0028] (1) Inoculate a single colony of ATCC 25922 or ATCC 29213 in a centrifuge tube containing 4 mL of MH broth, place in a shaker at 37°C at 180 rpm for 4 hours, and then remove the centrifuge tube;

[0029] (2) Use MH broth to dilute the incubated ATCC25922 and ATCC29213 100 times, about 10 6 CFU / mL, reserve;

[0030] (3) Take a sterile 96-well plate, add 180 μL of MH broth medium to well 1, and add 100 μL of MH broth medium to wells 2-11;

[0031] (4) Add 20 μL of the original concentration of wintergreen essential oil / potassium persulfate to the first well, blow and mix evenly, then pipette 100 μL into the second well, and so on, pipette 100 μL into the tenth well and discard; at this time, the concentrations of wintergreen essential oil / potassium persulfate in each well are 10%, 5%, 2.5%, 1.25%, 0.625%, 0.3125%, 0.156%, 0.078%, 0.039%, and 0.0195%, respectively.

[0032] (5) 100 μL of diluted bacterial solution was added to wells 1 to 10. At this time, the concentrations of wintergreen essential oil / potassium persulfate in each well were 5%, 2.5%, 1.25%, 0.625%, 0.3125%, 0.156%, 0.078%, 0.039%, 0.0195%, and 0.01%, respectively. 100 μL of bacterial solution was added to well 11, and 200 μL of MH broth was added to well 12;

[0033] (6) Repeat steps (3) to (5) three times in parallel;

[0034] (7) Place the inoculated 96-well plate in a 37-degree incubator and incubate for 16-18 hours. Then, pipette 10 μL of 10 mM / L resin azure into the wells. The resin azure concentration is 0.1 mM / L. After incubation for 2 hours, read the results.

[0035] The results are shown in Table 2. The MIC value of wintergreen essential oil against ATCC25922 is 0.156%, and the MIC value against ATCC29213 is 0.078%. At this concentration, the growth of bacteria is inhibited. The MIC value of potassium persulfate against ATCC25922 is 2.5%, and the MIC value against ATCC29213 is 5%. At this concentration, the growth of bacteria is inhibited. Since the action time of potassium persulfate and wintergreen essential oil is short, 0.5% and 1% wintergreen essential oil and 0.01% potassium persulfate are selected for combination.

[0036] Table 1 MIC of wintergreen essential oil and potassium persulfate against ATCC 25922 and ATCC 29213 strains

[0037]

[0038] Example 2: Bactericidal effect of wintergreen essential oil / potassium persulfate on ATCC 25922 and ATCC 29213 strains after 1 hour of action

[0039] 1. Experimental materials:

[0040] (1) Potassium persulfate and wintergreen essential oil.

[0041] (2) Experimental culture medium: The autoclaved LB agar medium (purchased from Guangdong Huankai Microbiological Technology Co., Ltd.) was cooled to 55°C, 20 mL was taken into a sterile culture dish with a pipette, and naturally dried for 30 minutes to prepare LB.

[0042] agar medium and MacConkey agar medium.

[0043] (3) Escherichia coli standard strains ATCC25922 and ATCC29213 (laboratory collection).

[0044] 2. Preparation before the test:

[0045] (1) The standard strain of Escherichia coli ATCC25922 and the standard strain of Staphylococcus aureus were cultured on LB agar medium until they reached an appropriate size.

[0046] (2) Dilute potassium persulfate 1000 times with physiological saline to a concentration of 0.1% for later use.

[0047] (3) Dilute wintergreen essential oil to 5% and 10% concentrations with 1% DMSO and set aside.

[0048] 3. Wintergreen essential oil / potassium persulfate bactericidal effect evaluation experiment:

[0049] (1) Inoculate a single colony of ATCC 25922 or ATCC 29213 in a centrifuge tube containing 4 mL of MH broth, place in a shaker at 37°C at 180 rpm for 4 hours, and then remove the centrifuge tube;

[0050] (2) Place the centrifuge tube in a centrifuge and centrifuge at 5000 rpm for 8 min. Pour off the supernatant and add an equal volume of saline to resuspend the tube so that the final bacterial count is approximately 10 8 CFU / mL;

[0051] (3) Set up different treatment groups, mix well and add to 6-well plates and incubate at 37°C for 1 h

[0052] ①Blank control group: 900 μL bacterial solution + 100 μL normal saline;

[0053] ②Potassium persulfate treatment group: 900 μL bacterial solution + 100 μL of the above diluted 0.1% potassium persulfate;

[0054] ③ Wintergreen essential oil treatment group: 900 μL bacterial solution + 100 μL of the above-diluted 5% / 10% wintergreen essential oil.

[0055] (4) After the incubation, 100 μL of bacterial solution was added to a 2 mL centrifuge tube containing 900 μL of 0.85% saline for gradient dilution. After dilution, 25 μL was dropped on MH agar medium and incubated in a 37°C incubator for 16-18 h. The cells were counted and the experimental results were statistically analyzed after three biological replicates.

[0056] (5) The results are shown in Table 2. The number of bacteria in the selected concentration of wintergreen essential oil and potassium persulfate treatment group is not much different from the number of bacteria in the blank control group, indicating that the selected concentration of wintergreen essential oil and potassium persulfate cannot produce a significant antibacterial effect on ATCC25922. The results are shown in Table 3. The number of bacteria in the selected concentration of wintergreen essential oil and potassium persulfate treatment group is not much different from the number of bacteria in the blank control group, indicating that the selected concentration of wintergreen essential oil and potassium persulfate cannot produce a significant antibacterial effect on ATCC29213.

[0057] Table 2 Bactericidal effect of holly essential oil and potassium persulfate on ATCC 25922 strain

[0058]

[0059] Table 3 Bactericidal effect of holly essential oil and potassium persulfate on ATCC29213 strain

[0060]

[0061] Example 3: Bactericidal effect of wintergreen essential oil combined with potassium persulfate for 1 h on ATCC 25922 and ATCC 29213 strains

[0062] 1. Experimental materials:

[0063] (1) Potassium persulfate and wintergreen essential oil.

[0064] (2) Experimental culture medium: The autoclaved LB agar medium (purchased from Guangdong Huankai Microbiological Technology Co., Ltd.) was cooled to 55°C, 20 mL was taken into a sterile culture dish using a pipette, and naturally dried for 30 min to prepare LB agar medium and MacConkey agar medium.

[0065] (3) Escherichia coli standard strains ATCC25922 and ATCC29213 (laboratory collection).

[0066] 2. Preparation before the test:

[0067] (1) The standard strain of Escherichia coli ATCC25922 and the standard strain of Staphylococcus aureus were cultured on LB agar medium until they reached an appropriate size.

[0068] (2) Dilute potassium persulfate 1000 times with physiological saline to a concentration of 0.1% for later use.

[0069] (3) Dilute wintergreen essential oil to 5% and 10% concentrations with 1% DMSO and set aside.

[0070] 3. Experimental evaluation of the combined bactericidal effect of wintergreen essential oil and potassium persulfate:

[0071] (1) Inoculate a single colony of ATCC 25922 or ATCC 29213 in a centrifuge tube containing 4 mL of MH broth, place in a shaker at 37°C at 180 rpm for 4 hours, and then remove the centrifuge tube;

[0072] (2) Place the centrifuge tube in a centrifuge and centrifuge at 5000 rpm for 8 min. Pour off the supernatant and add an equal volume of saline to resuspend the tube so that the final bacterial count is approximately 10 8 CFU / mL;

[0073] (3) Set up different treatment groups, mix well and add to 6-well plates and incubate at 37°C for 1 h

[0074] ①Blank control group: 800 μL bacterial solution + 200 μL normal saline;

[0075] ②Potassium persulfate treatment group: 800 μL bacterial solution + 100 μL normal saline + 100 μL of the above diluted 0.1% potassium persulfate;

[0076] ③ 0.5% wintergreen essential oil treatment group: 800 μL bacterial solution + 100 μL normal saline + 100 μL of the above diluted 5% wintergreen essential oil;

[0077] ④1% wintergreen essential oil treatment group: 800 μL bacterial solution + 100 μL normal saline + 100 μL of the above diluted 10% wintergreen essential oil;

[0078] ⑤ Potassium persulfate + 0.5% wintergreen essential oil treatment group: 800 μL bacterial solution + 100 μL of the above diluted 0.1% potassium persulfate + 100 μL of the above diluted 5% wintergreen essential oil;

[0079] ⑥Potassium persulfate+1% wintergreen essential oil treatment group: 800 μL bacterial solution+100 μL of the above diluted 0.1% potassium persulfate+100 μL of the above diluted 10% wintergreen essential oil.

[0080] (4) After the incubation, 100 μL of bacterial solution was added to a 2 mL centrifuge tube containing 900 μL of 0.85% saline for gradient dilution. After dilution, 25 μL was dropped on MH agar medium and incubated in a 37°C incubator for 16-18 h. The cells were counted and the experimental results were statistically analyzed after three biological replicates.

[0081] (5) The antibacterial effect of wintergreen essential oil and potassium persulfate was weak. The use of 0.01% potassium persulfate alone reduced the bacterial count by 0.36 log compared with the control group, and the use of 0.5% and 1% wintergreen essential oil alone reduced the bacterial count by 0.07 log and 0.9 log compared with the control group. The combination of them had a significant synergistic antibacterial effect, and the plate count results showed that Escherichia coli ATCC25922 was completely killed; as shown in Table 5 and Figure 2 When the concentration of 0.01% potassium persulfate and 0.5% wintergreen essential oil were combined, the bactericidal effect on Staphylococcus aureus ATCC29213 was significantly improved compared with when used alone, and the bacterial count decreased by >1.5log. When the concentration of 0.01% potassium persulfate and 1% wintergreen essential oil were combined, the bactericidal effect on Staphylococcus aureus ATCC29213 was significantly improved compared with when used alone, and the bacterial count decreased by >4.5log.

[0082] Compared with traditional potassium persulfate disinfection, the present invention can kill bacteria more efficiently and reduce the potassium persulfate concentration used.

[0083] Table 4 Bactericidal effect of potassium persulfate and wintergreen essential oil on Escherichia coli ATCC 25922

[0084]

[0085] Table 5 Bactericidal effect of potassium persulfate and wintergreen essential oil on Staphylococcus aureus ATCC29213

[0086]

[0087] The above embodiments of the present invention are merely examples for clearly illustrating the technical solution of the present invention, and are not intended to limit the specific implementation methods of the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the claims of the present invention shall be included in the protection scope of the claims of the present invention.

Claims

1. A disinfection synergist, characterized in that: The composite disinfection composition is dissolved in a solvent to obtain the disinfection synergist; the composite disinfection composition consists of potassium persulfate and wintergreen essential oil; the concentration of the potassium persulfate is 0.01%-1%; the concentration of the wintergreen essential oil is 0.1%-2%.

2. The disinfection synergist according to claim 1, characterized in that: The solvent is water.

3. Use of the disinfection enhancer according to claim 1 in the preparation of products in the field of disinfection and purification.

4. The use according to claim 3, characterized in that: Disinfection and purification are used to disinfect Escherichia coli and Staphylococcus aureus.

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