Concentrated alkaline multienzyme cleaning agent and preparation method thereof

Abstract

The invention discloses a concentrated alkaline multienzyme cleaning agent and a preparation method thereof. The concentrated multi-enzyme cleaning agent comprises the following components: (1) a multi-enzyme mixed solution formed by mixing protease, lipase, amylase and cellulase, wherein the multi-enzyme mixed solution is a component A; (2) a solution formed by mixing a corrosion inhibitor, a stabilizer, a buffering agent, a surfactant and deionized water is a component B; wherein the buffering agent is glycine and boric acid. The concentrated alkaline multi-enzyme cleaning agent is high in cleaning capacity, has good cleaning capacity on various pollutants such as protein, a bacterial blood mixture and a biological membrane, is easy to store, can keep stable enzyme activity in a long time, is long in shelf life, and can be used for cleaning various pollutants such as the protein, the bacterial blood mixture and the biological membrane. The device can be widely applied to hospitals, third-party decontamination centers, scientific research laboratories, hotels, schools, families and other places.

Description

Technical Field

[0001] The invention relates to the field of chemistry, and in particular to a concentrated alkaline multi-enzyme cleaning agent and a preparation method thereof. Background Art

[0002] Multi-enzyme cleaning agents are suitable for cleaning medical devices. Surgical instruments have dirt attached to their surfaces, such as protein, blood, fat and some microorganisms. When these dirt dries on the surface of medical instruments, it is difficult to clean them in a short time. Biological enzymes such as proteases, amylases and lipases can enzymatically hydrolyze the dirt attached to medical instruments into substances that are easily soluble in surfactants, thereby completely removing them. However, pH has a great influence on the decomposition of dirt on medical instruments. Too high or too low pH can easily cause corrosion to medical instruments and is not conducive to the subsequent cleaning of medical instruments. In order to solve this problem, one approach is to use multiple enzymes under neutral conditions, so that the cleaning agent can not only decompose the dirt, but also will not contaminate the medical device, which is beneficial to subsequent cleaning and increases the safety of the product. However, multiple enzymes are prone to cause the activity of some enzymes to decrease, and the effective shelf life of the product is short. In addition, in order to achieve the cleaning effect, the common practice is to increase the types of components and the amount added, which not only leads to high costs and overly complicated preparation, but also is not conducive to industrial production. Another approach is a single enzyme cleaning agent. Although the enzyme activity of a single enzyme is improved, it cannot completely remove the dirt on the medical device, increasing the difficulty of the subsequent cleaning steps.

[0003] The short effective shelf life of multi-enzyme cleaning agents on the market, rapid decline in enzyme activity, high production costs, and complex components are the main factors restricting the development of multi-enzyme cleaning agents. Therefore, it is urgent to develop a multi-enzyme cleaning agent with low cost, easy production and preparation, strong cleaning ability, long shelf life, and stable enzyme activity. Summary of the invention

[0004] The purpose of the present invention is to provide a multi-enzyme cleaning agent with few components, easy production and preparation, strong cleaning ability, long shelf life and stable enzyme activity.

[0005] In a first aspect of the present invention, a concentrated multi-enzyme cleaning agent is provided, wherein the multi-enzyme cleaning agent comprises the following components:

[0006] (1) a multi-enzyme mixture formed by protease, lipase, amylase and cellulase is component A;

[0007] (2) a solution formed by mixing a corrosion inhibitor, a stabilizer, a buffer, a surfactant and deionized water is component B; and the buffer is glycine and boric acid;

[0008] Furthermore, component A and component B are further mixed to form the concentrated multi-enzyme cleaning agent;

[0009] The concentrated multi-enzyme cleaning agent contains the following ingredients: protease 0.3-5IU / mL; lipase 0.5-5IU / mL; amylase 0.2-5IU / mL; cellulase 0.2-5IU / mL; corrosion inhibitor 3-10w / w%; stabilizer 20-30w / w%; buffer 1-2mol / L; surfactant 0.05-1.0mol / L; and the balance is deionized water.

[0010] In another preferred embodiment, the amount of deionized water used is 40%-80% (W / W).

[0011] In another preferred embodiment, the volume ratio of component A to component B is (1-5):(1-5).

[0012] In another preferred embodiment, the molar ratio of glycine to boric acid in the buffer is 2:1 to 20:1.

[0013] In another preferred embodiment, in the concentrated multi-enzyme cleaning agent, the glycine is 0.5-1.9 mol / L, and the boric acid is 0.04-0.9 mol / L.

[0014] In another preferred embodiment, the corrosion inhibitor is triethanolamine, sodium silicate, or a combination thereof.

[0015] In another preferred embodiment, the stabilizer is glycerol.

[0016] In another preferred embodiment, the surfactant is: AEO-9 (fatty alcohol polyoxyethylene ether), decyl glucoside APG0810, Brij 35 (lauryl alcohol polyoxyethylene ether), or a combination thereof.

[0017] In another preferred embodiment, the concentrated multi-enzyme cleaning agent further comprises a chelating agent.

[0018] In another preferred embodiment, the dosage of the chelating agent is 0.05-1.0 (mol / L).

[0019] In another preferred embodiment, the chelating agent is EDTA-sodium.

[0020] In another preferred embodiment, the concentrated multi-enzyme cleaning agent contains the following components: protease 0.3-5IU / mL; lipase 0.5-5IU / mL; amylase 0.2-5IU / mL; cellulase 0.2-5IU / mL; triethanol and / or sodium silicate 3-10w / w%; glycerol 20-30w / w%; glycine and boric acid 1-2mol / L; AEO-9 and / or decyl glucoside APG 0810 and / or Brij 35 0.05-1.0mol / L; and the balance is deionized water.

[0021] In another preferred embodiment, the concentrated multi-enzyme cleaning agent contains the following components: protease 0.3-5IU / mL; lipase 0.5-5IU / mL; amylase 0.2-5IU / mL; cellulase 0.2-5IU / mL; triethanol and / or sodium silicate 3-10w / w%; glycerol 20-30w / w%; glycine and boric acid 1-2mol / L; AEO-9 and / or decyl glucoside APG 0810 and / or Brij 350.05-1.0mol / L; EDTA-sodium 0.05-1.0 (mol / L); and the balance is deionized water.

[0022] In another preferred embodiment, the pH of the concentrated multi-enzyme cleaning agent is 7.1-8.8.

[0023] In another preferred embodiment, the pH of the concentrated multi-enzyme cleaning agent is 7.6-8.6, more preferably 7.8-8.6.

[0024] In another preferred embodiment, the pH adjuster of the concentrated multi-enzyme cleaning agent is HCl or NaOH.

[0025] In another preferred embodiment, the protease is a protease from Bacillus, the lipase is a lipase from Candida rugosa, and / or the amylase is an amylase from Bacillus.

[0026] In a second aspect of the present invention, a method for cleaning a utensil is provided, comprising the steps of: (a) providing a concentrated multi-enzyme cleaning agent as described in the first aspect of the present invention, and diluting the concentrated multi-enzyme cleaning agent by 10-600 times to obtain a diluted multi-enzyme cleaning agent;

[0027] (b) cleaning the utensil with the diluted multi-enzyme cleaning agent.

[0028] The concentrated multi-enzyme cleaning agent as described in the first aspect of the present invention is used to clean the utensil after diluting it 10-600 times.

[0029] In another preferred embodiment, the dilution factor of the cleaning agent is 200-400 times.

[0030] In another preferred embodiment, the dilution factor of the cleaning agent is 200 times.

[0031] In another preferred embodiment, the starch removal rate of the cleaning agent is greater than 95%. Preferably, the starch removal rate of the cleaning agent is greater than or equal to 99%.

[0032] In another preferred embodiment, the protein removal rate of the cleaning agent is greater than 95%. Preferably, the protein removal rate of the cleaning agent is greater than or equal to 98%.

[0033] In another preferred embodiment, the fat removal rate of the cleaning agent is greater than 90%. Preferably, the fat removal rate of the cleaning agent is greater than or equal to 95%.

[0034] In another preferred embodiment, the instrument is selected from the following group: surgical instruments, laboratory instruments, or a combination thereof.

[0035] In another preferred embodiment, the device contains pollutants selected from the group consisting of starch pollutants, oil pollutant preparations, protein pollutants, bacterium-blood mixed pollutants, microbial biofilms, or combinations thereof.

[0036] In another preferred embodiment, the device contains the following pollutants: microbial biofilm.

[0037] In another preferred embodiment, the concentrated multi-enzyme cleaning agent is a cleaning agent stock solution stored at a certain temperature T (such as room temperature, or 4-37° C.) for a period of time t (such as ≥6 months, ≥9 months, or ≥12 months).

[0038] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 Diagram of the biofilm removal experimental setup. DETAILED DESCRIPTION

[0040] The inventors have conducted extensive and in-depth research, and through a large number of screenings and tests, have provided for the first time a concentrated alkaline multi-enzyme cleaning agent with excellent performance. The concentrated alkaline multi-enzyme cleaning agent of the present invention not only has a strong cleaning ability, but also has excellent cleaning ability for various pollutants such as proteins, bacterium-blood mixtures, etc., and also has a very high removal rate for microorganisms (such as bacteria) in stubborn microbial biofilms. In addition, the concentrated alkaline multi-enzyme cleaning agent of the present invention also has the advantages of being easy to store and stable, and the enzyme activity, decontamination ability and bacteria removal performance can remain stable for a long time, and the shelf life is long. The present invention was completed on this basis.

[0041] Concentrated alkaline multi-enzyme cleaner

[0042] As used herein, the terms "concentrated multi-enzyme cleaning agent of the present invention", "concentrated alkaline multi-enzyme cleaning agent of the present invention", "alkaline concentrated multi-enzyme cleaning agent of the present invention", or "concentrated cleaning agent of the present invention" are used interchangeably to refer to the alkaline concentrated multi-enzyme cleaning agent described in the first aspect of the present invention. It should be understood that the concentrated cleaning agent of the present invention may contain trace amounts of impurities that do not interfere with cleaning performance.

[0043] Typically, the concentrated multi-enzyme cleaning agent of the present invention is obtained by mixing component A and component B.

[0044] Typically, based on the active content of the final product solution, component A:

[0045] include:

[0046] Protease 0.3-5 IU / mL;

[0047] Lipase 0.5-5 IU / mL;

[0048] Amylase 0.2-5 IU / mL;

[0049] Cellulase 0.2-5 IU / mL;

[0050] Preferably,

[0051] The protease is derived from Bacillus;

[0052] The lipase was derived from Candida rugosa;

[0053] Amylase is derived from Bacillus.

[0054] Component B contains the following components based on the mass percentage of the final finished product solution:

[0055] Corrosion inhibitor: one or more of triethanolamine or sodium silicate: 3-10w / w%;

[0056] Stabilizer: glycerol: 20-30w / w%;

[0057] Buffer: glycine and boric acid, preferably the molar ratio of glycine to boric acid is 2:1-20:1, and the total amount of the two is 1-2 (mol / L);

[0058] Surfactant: one or more of AEO-9 (fatty alcohol polyoxyethylene ether), decyl glucoside APG 0810, or Brij35 (lauryl alcohol polyoxyethylene ether); the total content of the surfactant is usually 0.05-1.0 mol / L.

[0059] Preferably, a chelating agent is also included, wherein the chelating agent is EDTA-sodium, and the amount of the chelating agent is 0.05-1.0 mol / L;

[0060] The balance is deionized water. Preferably, the content of deionized water is 40%-80% (W / W).

[0061] The concentrated multi-enzyme cleaning agent of the present invention is weakly alkaline, and its pH is usually 7.1-8.8, preferably 7.6-8.6, and more preferably 7.8-8.6. Usually, HCl or NaOH can be used to adjust the pH.

[0062] Preparation method of multi-enzyme cleaning agent

[0063] The invention also provides a preparation method of the concentrated multi-enzyme cleaning agent.

[0064] A preferred method comprises the following steps:

[0065] Step 1: Add appropriate amount of deionized water, appropriate amount of propylene glycol, and the formulated amount of protease, lipase, and amylase, stir, and filter to obtain component A.

[0066] Step 2: Add appropriate amount of deionized water, add the formulated amount of corrosion inhibitor, stabilizer, buffer, surfactant, chelating agent, add appropriate amount of deionized water, stir until completely dissolved, and obtain component B after filtering.

[0067] Step 3: Component A and component B are mixed in a certain proportion to obtain a concentrated multi-enzyme cleaning solution.

[0068] Step 4: Use NaOH or HCl to adjust the pH of the concentrated multi-enzyme cleaning solution obtained in step 3.

[0069] Preferably, in step 1 and step 2, the filtration is performed by 0.45 μm filter membrane filtration.

[0070] Preferably, in the step 1, the stirring speed is 5-100 rpm, and the dissolving temperature is 15°C-28°C.

[0071] Preferably, in step 1 and step 2, the filtration is performed by 0.45 μm filter membrane filtration.

[0072] Preferably, the volume ratio of component A to component B is (1-5):(1-5).

[0073] Biofilm

[0074] Biofilms are a mixture of bacteria, fungi, protozoa and / or algae. They are a complex of microbial communities that usually form at the boundary layer between the gas and liquid phases, such as pipes, cooling towers, drains and other water transport systems. Microorganisms can synthesize and secrete a protective matrix through which the biofilm is firmly attached to the inside of the pipe. They are usually seen as a layer of slime or a coating with different thickness, color and consistency. The characteristic biofilm slime matrix is ​​composed of extracellular polymers (EPS) produced by microorganisms, which form a hydrogel with water.

[0075] Biofilms are in a constant state of flux. They may consist of a single bacterial or fungal population, but in most cases they consist of multiple populations, i.e. a diverse collection of bacteria. Basically, biofilms can be described as bacteria hiding in a thick, slimy protective layer made of sugars and proteins. The biofilm protective layer protects the microorganisms from external hazards. Therefore, biofilms that grow inside pipes are very resistant and difficult to completely remove.

[0076] Luminal surface culture method

[0077] One method of evaluating the effectiveness of biofilm removal is to use the luminal surface culture method. This method involves collecting samples from the inner surface of the pipe after treatment and then culturing them under laboratory conditions to evaluate the residual biofilm. By comparing the number of microorganisms in the samples before and after treatment, the effectiveness of biofilm removal can be quantified. This method can help determine the efficiency of the treatment method and whether the expected cleaning standards have been achieved.

[0078] the term

[0079] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0080] As used herein, when used in reference to a specific recited numerical value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0081] As used herein, the term "comprising" or "including (comprising)" may be open, semi-closed and closed. In other words, the term also includes "consisting essentially of" or "consisting of".

[0082] As used herein, the term "room temperature" or "normal temperature" refers to a temperature of 4-40°C, preferably, 25±5°C.

[0083] As used herein, the term "AEO-9" refers to fatty alcohol polyoxyethylene ether.

[0084] As used herein, the term "APG 0810" refers to decyl glucoside.

[0085] As used herein, the term "Brij 35" refers to polyoxyethylene lauryl ether.

[0086] As used herein, the terms "concentrated multi-enzyme cleaning agent", "multi-enzyme cleaning solution stock solution" and "multi-enzyme cleaning solution stock solution dilution" are used interchangeably, and all refer to a mixed solution including component A and component B.

[0087] As used herein, the term "component A" refers to a mixed solution comprising formulated amounts of protease, lipase, amylase and cellulase.

[0088] As used herein, the term "component B" refers to a mixed solution including a corrosion inhibitor, a stabilizer, a buffer, a surfactant and deionized water.

[0089] The main advantages of the present invention include:

[0090] 1. The concentrated multi-enzyme cleaning agent of the present invention has strong cleaning ability and has excellent cleaning ability for various pollutants such as starch pollutants, grease pollutants, protein pollutants, bacterium-blood mixed pollutants and biofilms, which is better than existing cleaning agents on the market and can maintain strong cleaning ability after a large amount of dilution.

[0091] 2. The concentrated multi-enzyme cleaning agent of the present invention has low corrosiveness. While maintaining strong cleaning ability, it does not corrode medical equipment or experimental equipment that need to be cleaned. It is safer and suitable for use in public places such as schools.

[0092] 3. The concentrated multi-enzyme cleaning agent of the present invention has good stability. Unlike the existing multi-enzyme cleaning agents, the enzyme activity decreases slowly and the product has a long effective shelf life.

[0093] 4. The concentrated multi-enzyme cleaning agent of the present invention has simple components. Unlike the prior art formulas that have more than ten components, the simplest formula contains only nine components, which greatly reduces the preparation cost, time and operation difficulty.

[0094] The present invention will be further described below in conjunction with specific implementation. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0095] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0096] Example 1 Alkaline concentrated multi-enzyme cleaning agent and its preparation

[0097] The preparation method of the concentrated multi-enzyme cleaning agent is carried out using the formula shown in Table 1. The preparation method includes the following steps:

[0098] Add appropriate amount of deionized water, add appropriate amount of glycerol, add the formulated amount of protease, lipase, and amylase (see Table 1), stir, and filter through a 0.45 μm filter membrane to obtain component A.

[0099] Add appropriate amount of deionized water, add the formulated amount of stabilizer, corrosion inhibitor, buffer, surfactant, chelating agent (see Table 1), add appropriate amount of deionized water, stir until completely dissolved, and filter through 0.45 μm to obtain component B.

[0100] Component A and component B are mixed in a certain proportion to obtain a multi-enzyme cleaning solution, and then the pH of the multi-enzyme cleaning solution is adjusted to 7.1-8.8 with NaOH or HCl.

[0101] The formula of the multi-enzyme cleaning solution is shown in Table 1.

[0102] Table 1 Recipe

[0103]

[0104] Note: “-” in the table means none.

[0105] Example 2 Corrosion Test

[0106] In Example 1, the multi-enzyme cleaning solution of formula 1-7 was diluted 200 times and poured into a 50 mL glass jar with a lid. Four commercially available metal sheets: carbon steel GB 700-65, aluminum GB 1173-74, copper GB 2060-80, and stainless steel GB1220-75, with a diameter of 24±0.1 mm and a thickness of 1.0 mm, were placed in the cleaning solution and immersed for 72 hours. The weight of each metal sheet before and after treatment was weighed, and the weight loss value was calculated.

[0107] Observed with the naked eye, the cleaning solutions of formulas 1-7 of the present invention are non-corrosive. Experimental data show that the corrosion rates of carbon steel GB 700-65, aluminum GB 1173-74, copper GB 2060-80, and stainless steel GB1220-75 are less than or equal to 0.0011 mm / a, 0.0025 mm / a, 0.0012 mm / a, and 0.0090 mm / a, respectively, which are much less than 0.0100 mm / a (the criterion for non-corrosion). It can be concluded that the cleaning agents of formulas 1-7 are non-corrosive and have a mild effect.

[0108] Comparative Examples 1 and 2 are severely corroded. The corrosion rates of carbon steel GB 700-65, aluminum GB 1173-74, copper GB 2060-80, and stainless steel GB1220-75 are respectively greater than 0.0263mm / a, 0.0155mm / a, 0.0173mm / a, and 0.0259mm / a. Compared with 0.0100mm / a (the judgment standard for basically no corrosion), it shows that the comparative examples are corrosive to a variety of materials and are highly irritating. The corrosion rates of commercially available products on carbon steel GB 700-65, aluminum GB 1173-74, copper GB 2060-80 and stainless steel GB1220-75 are 0.0057mm / a, 0.0084mm / a, 0.0037mm / a and 0.0106mm / a respectively. Compared with 0.0100mm / a (the judgment standard for basically no corrosion), it shows that the commercially available products have certain corrosiveness to stainless steel GB1220-75.

[0109] Example 3: Test of the decontamination effect on artificial simulated pollutants

[0110] Step 1: Preheat the oil at 60℃, take 2.4mL, add 2g BSA, 1g corn starch, add water to 20mL to obtain artificial simulated pollutants, add it dropwise on the STF cleaning effect test card, place the STF cleaning effect test card in a desiccator overnight, and weigh it as M0 using an analytical balance.

[0111] Step 2: Place the artificial simulated pollutant carrier in step 1, i.e., the STF cleaning effect test card, into a glass jar containing 300 ml of multi-enzyme cleaning agent (the stock solution was diluted 200 times), and wash at 40°C at a speed of 50 r / min for 5 minutes. Remove the glass jar, take out the carrier, dry it at 30°C for 2 hours, and weigh it as M1 using an analytical balance.

[0112] Step 3: Remove the artificial simulated pollutants remaining on the STF cleaning effect test card and weigh them as M2 using an analytical balance.

[0113] The cleaning effect is evaluated according to the following formula:

[0114] Removal efficiency R = 100%*(M0-M1) / (M0-M2)

[0115] Where: M0——mass of simulated pollutants before cleaning, unit: g

[0116] M1——The mass of simulated pollutants after cleaning with multi-enzyme cleaning agent, unit: g

[0117] M2——mass of artificially simulated pollutants after cleaning, unit: g

[0118] The cleaning effect is shown in Table 2.

[0119] Table 2 Cleaning effect of each formula on artificial simulated pollutants

[0120] formula Cleaning efficiency 1 99.49％ 2 98.99％ 3 ≥99.99％ 4 99.55％ 5 99.27％ 6 99.79％ 7 99.86％ Comparative Example 1 87.32％ Comparative Example 2 89.85％ Commercially available products 95.44％

[0121] As can be seen from Table 2, after the original solution of the multi-enzyme cleaning solution of the present invention is diluted 200 times, the cleaning efficiency of removing pollutants is about 99%, and the preferred formula can be ≥99.99%, which is far higher than the cleaning rate qualification standard (95%) of the STF cleaning effect test card. The cleaning efficiency of the comparative cleaning agents is less than 90%.

[0122] Example 4 Decontamination effect test of cleaning agent on fat, protein and starch

[0123] The test was conducted with reference to the experimental method for evaluating the removal effect of fat, protein and starch by medical cleaning agents in Appendix E of "Hygiene Requirements for Medical Cleaning Agents" T / WSJT 002-2019.

[0124] Preparation of starch pollutants: 3 g of starch and 0.5 g of wallpaper paste were dissolved in an appropriate amount of water, stirred thoroughly and water was added to 100 mL to obtain simulated starch pollutants.

[0125] Preparation of protein pollutants: 3 g of bovine serum albumin and 0.5 g of wallpaper paste were dissolved in an appropriate amount of water, stirred thoroughly and water was added to 100 mL to obtain simulated protein pollutants.

[0126] Preparation of oil contaminants: Preheat edible lard in a 45°C water bath until the lard melts into a liquid state.

[0127] To prepare starch and protein pollution slices: take defatted and air-dried polytetrafluoroethylene, insert the polytetrafluoroethylene into the pollutant, slowly take it out, dry it and weigh it.

[0128] To make a grease contaminated sheet: take the defatted and dried polytetrafluoroethylene, use a pipette to absorb 20uL of lard and evenly spread it on the polytetrafluoroethylene, and then weigh it.

[0129] Cleaning of pollutants by cleaning agent: dilute the original multi-enzyme cleaning solution in Example 1 200 times and place it in an open glass bottle, put the polluted sheets into the dilution solution in the open glass bottle, rotate and wash at 45°C at a speed of 50r / min for 5 minutes, remove the carrier sheet, put it in a dryer, and weigh it after 2 hours. Evaluate the removal effect of fat, protein and starch.

[0130] The experimental results are shown in Table 3 below.

[0131] Table 3 Effects of cleaning agents and commercial products on fat, protein and starch removal

[0132] serial number Starch removal (%) Protein removal (%) Fat removed (%) Recipe 1 99.77 99.67 95.85 Recipe 2 99.81 ≥99.99 95.88 Recipe 3 99.85 99.49 95.75 Recipe 4 99.95 99.58 95.98 Recipe 5 99.92 98.89 96.82 Recipe 6 99.85 99.94 98.96 Recipe 7 99.78 99.88 95.86 Comparative Example 1 65.65 91.67 78.85 Comparative Example 2 61.45 90.32 85.63 Commercially available products 75.31 95.98 85.57

[0133] From the results in Table 3, it can be seen that after the multi-enzyme cleaning solution of the present invention is diluted 200 times, the cleaning efficiency of starch, protein and fat is higher than that of the comparative example and the commercially available product, and the disadvantage of low starch and / or fat removal rate of the general products on the market is improved, and the starch and fat removal rate is greatly improved, which helps to fill the market gap. Specifically, the cleaning efficiency of starch is greater than 99%, while the cleaning efficiency of the cleaning agent of the comparative example is less than 65.65%, and the cleaning efficiency of the commercially available product is 75.31%; the cleaning efficiency of protein is about 99%, and the preferred formula can be ≥99.99%, while the cleaning efficiency of the cleaning agent of the comparative example is less than 91.67%, and the cleaning efficiency of the commercially available product is 95.98%; the cleaning efficiency of fat is greater than 95%, while the cleaning efficiency of the cleaning agent of the comparative example is less than 71.63%, and the cleaning efficiency of the commercially available product is 77.57%.

[0134] Example 5: Effect of cleaning agent on removal of mixed pollutants of bacterium and blood

[0135] Preparation of bacterium-blood mixture: 0.35 mL of cultured enterococci, 9.5 mL of heparinized blood, and 0.15 mL of protamine hydrochloride (concentration of 4 mg / mL) were mixed, and the blood coagulated within 20 minutes to obtain the bacterium-blood mixture.

[0136] Preparation of carrier: The degreased and sterilized stainless steel screws were completely immersed in the test pollutant for 1 min, taken out, and dried at 36°C until the blood coagulated.

[0137] With reference to the test method for the removal effect of a mixture of blood and bacteria in Appendix C of "Hygiene Requirements for Medical Cleaning Agents" T / WSJT 002-2019, the cleaning agent of formula 1-7 described in the present invention is used to clean the contaminated carrier: 1-2 screws are placed in a diluent containing a 300-fold dilution of the multi-enzyme cleaning solution, and the screws are rotated and washed at a rotation speed of 45 r / min for 5 minutes. The screws are taken out and placed in a conventional eluent, and the cleaning effect of the cleaning agent on the contaminated carrier is evaluated by bacterial counting and ATP content determination.

[0138] After analyzing the experimental results, the cleaning agents of formula 1-7 in the experiment have a bacteria removal rate of more than 99.99%. The bacteria removal rates of comparative examples 1 and 2 are 99.63% and 99.24% respectively, which is higher than the 99.70% bacteria removal rate of commercial products. When tested with the Jerry test paper, formula 1-7, comparative examples 1-2 and commercial products were all negative, indicating that the blood has been cleaned.

[0139] Example 6 Determination of biofilm removal effect

[0140] Refer to the tube inner surface culture method in the biofilm removal effect determination method in Appendix F of "Medical Cleaning Agent Hygiene Requirements" T / WSJT 002-2019 for testing. Figure 1 As shown, an additional liquid storage system is added to the pipeline, and a polycarbonate sheet is placed in the bottle as a biofilm carrier. During the circulating culture, the culture liquid level is made to cover the polycarbonate sheet.

[0141] The experimental results are shown in Table 4 below.

[0142] Table 4 Bacterial removal rate and ATP content reduction rate in biofilm

[0143] serial number Bacteria removal rate (%) ATP content decrease rate (%) Recipe 1 99.92 99.91 Recipe 2 99.98 99.93 Recipe 3 ≥99.99 ≥99.99 Recipe 4 ≥99.99 ≥99.99 Recipe 5 99.96 99.97 Recipe 6 ≥99.99 ≥99.99 Recipe 7 99.88 99.87 Comparative Example 1 94.54 96.75 Comparative Example 2 95.31 94.63 Commercially available products 96.76 95.74

[0144] From this table, it can be seen that after the original solution of the multi-enzyme cleaning solution of the present invention is diluted 300 times, the bacteria removal rate and ATP content reduction rate in the biofilm are still better than those of the commercially available products and the comparative examples.

[0145] Example 7 Determination of α-amylase stability in cleaning solution

[0146] Referring to the test method for enzyme activity in Appendix B of "Hygiene Requirements for Medical Cleaning Agents" T / WSJT 002-2019, α-amylase activity was measured at three time points: ① within 1 hour after preparation; ② after storage at 37°C for 4 months; and ③ after storage at room temperature for 12 months.

[0147] The protease activity measured within 1 hour after the preparation was taken as the benchmark enzyme activity and was set to 100. The α-amylase activity measured at other time points was compared with the enzyme activity at the benchmark point. The experimental results are shown in Table 5 below.

[0148] Table 5 α-amylase activity (%)

[0149] serial number Preparation 1 hour Store at 37℃ for 4 months Store at room temperature for 12 months Recipe 1 100 98.91 98.93 Recipe 2 100 98.57 98.86 Recipe 3 100 99.83 99.84 Recipe 4 100 98.88 99.31 Recipe 5 100 98.35 98.42 Recipe 6 100 98.84 98.98 Recipe 7 100 98.74 98.92 Comparative Example 1 100 87.28 87.43 Comparative Example 2 100 81.54 82.21

[0150] As can be seen from Table 5, after the multi-enzyme cleaning agent of the present invention is stored at 37°C for 4 months, the enzyme activity of α-amylase maintains an extremely high enzyme activity value, which is higher than that of the comparative example. After long-term storage, i.e., storage at room temperature for 12 months, the activity of α-amylase is above 98.36% before storage, while the enzyme activity of the comparative example is less than 87.43% before storage.

[0151] Example 8 Determination of protease stability in cleaning solution

[0152] Referring to the test method for enzyme activity in Appendix B of "Hygiene Requirements for Medical Cleaning Agents" T / WSJT 002-2019, the protease activity was measured at three time points: ① 1 hour after preparation; ② 4 months of storage at 37°C; and ③ 12 months of storage at room temperature.

[0153] The protease activity measured within 1 hour after the preparation was taken as the benchmark enzyme activity and was set to 100. The protease activities measured at other time points were compared with the enzyme activity at the benchmark point. The experimental results are shown in Table 6.

[0154] Table 6 Protease activity (%)

[0155] serial number Preparation 1 hour Store at 37℃ for 4 months Store at room temperature for 12 months Recipe 1 100 98.33 98.53 Recipe 2 100 98.31 98.68 Recipe 3 100 98.85 98.01 Recipe 4 100 98.47 98.43 Recipe 5 100 98.59 98.98 Recipe 6 100 98.94 98.77 Recipe 7 100 98.59 98.48 Comparative Example 1 100 85.96 84.03 Comparative Example 2 100 85.53 83.94 Commercially available products 100 94.12 94.52

[0156] It can be seen from Table 6 that after the multi-enzyme cleaning agent of the present invention is stored at 37° C. for 4 months, the enzyme activity of the protease can also maintain extremely high enzyme activity values, which are higher than those of the comparative example and commercially available products.

[0157] Long-term storage, i.e. storage at room temperature for 12 months, has a protease activity of more than 98.33% before storage, while the activity of the control group enzymes is 84.03% and 83.94% before storage, and the activity of the commercially available product enzymes is 94.52% before storage. This patented formula exhibits superior stability.

[0158] Example 9 Determination of lipase stability in cleaning solution

[0159] Referring to the test method for enzyme activity in Appendix B of "Hygiene Requirements for Medical Cleaning Agents" T / WSJT 002-2019, lipase activity was measured at three time points: ① 1 hour after preparation; ② 4 months of storage at 37°C; and ③ 12 months of storage at room temperature.

[0160] The lipase activity measured within 1 hour after the preparation was taken as the benchmark value of 100, and the lipase activity measured at other time points was compared with the enzyme activity at the benchmark point.

[0161] The experimental results are shown in Table 7.

[0162] Table 7 Lipase activity (%)

[0163]

[0164]

[0165] From the table data, it can be seen that after the multi-enzyme cleaning agent of the present invention is stored at 37° C. for 4 months, the lipase activity is 91.33%, which is significantly higher than that of the comparative example and the commercially available product.

[0166] After long-term storage, i.e. storage at room temperature for 12 months, the lipase activity was above 98.64% before storage, while the enzyme activity of the control group was less than 90.35% before storage, and the enzyme activity of the commercially available product was 97.21% before storage. The patented formula exhibits excellent stability.

[0167] Example 10 Determination of cellulase stability in cleaning solution

[0168] Using a cellulase activity detection kit, the cellulase activity was measured at three time points: ① within 1 hour after preparation; ② after storage at 37°C for 4 months; and ③ after storage at room temperature for 12 months.

[0169] The cellulase activity measured within 1 hour after the preparation was set as 100, and the cellulase activity measured at other time points was compared with the enzyme activity at the reference point. The experimental results are shown in Table 8.

[0170] Table 8 Cellulase activity (%)

[0171] serial number Preparation 1 hour Store at 37℃ for 4 months Store at room temperature for 12 months Recipe 1 100 97.93 98.26 Recipe 2 100 97.69 98.61 Recipe 3 100 98.73 98.30 Recipe 4 100 98.14 98.55 Recipe 5 100 97.71 98.43 Recipe 6 100 98.96 98.33 Recipe 7 100 98.66 98.87 Comparative Example 1 100 81.56 80.33 Comparative Example 2 100 85.18 88.06

[0172] From the results in the table, it can be seen that after the multi-enzyme cleaning agent of the present invention is stored at 37° C. for 4 months, the cellulase activity is 97.69%, which is significantly higher than that of the comparative example.

[0173] Moreover, after long-term storage, i.e. storage at room temperature for 12 months, the cellulase activity was above 98.33% of that before storage, while the activity of the comparative enzyme was less than 76.33% before storage. The formula of the present invention exhibits excellent stability.

[0174] In summary, the multi-enzyme cleaning agent described in the present invention has excellent cleaning effect, low corrosivity, mild action, simple formula components and few compound ingredients. In addition, it has the outstanding effect of good multi-enzyme stability, among which α-amylase, protease, lipase and cellulase can maintain excellent stability and can show significant enzyme activity values ​​even after long-term storage, which is significantly better than existing commercially available products and can be widely used in the market.

[0175] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A concentrated multi-enzyme cleaning agent, characterized in that: The multi-enzyme cleaning agent comprises the following components: (1) a multi-enzyme mixture formed by mixing protease, lipase, amylase and cellulase is component A; (2) a solution formed by mixing a corrosion inhibitor, a stabilizer, a buffer, a surfactant and deionized water is component B; and the buffer is glycine and boric acid; Furthermore, component A and component B are further mixed to form the concentrated multi-enzyme cleaning agent; The multi-enzyme cleaning agent contains the following ingredients: protease 0.3-5IU / mL; lipase 0.5-5IU / mL; amylase 0.2-5IU / mL; cellulase 0.2-5IU / mL; corrosion inhibitor 3-10w / w%; stabilizer 20-30w / w%; buffer 1-2mol / L; surfactant 0.05-1.0mol / L; and the balance is deionized water.

2. The concentrated multi-enzyme cleaning agent according to claim 1, characterized in that The molar ratio of glycine to boric acid in the buffer is 2:1 to 20:

1.

3. The concentrated multi-enzyme cleaning agent according to claim 1, characterized in that In the multi-enzyme cleaning agent, glycine is 0.5-1.9 mol / L, and boric acid is 0.04-0.9 mol / L.

4. The concentrated multi-enzyme cleaning agent according to claim 1, characterized in that The corrosion inhibitor is triethanolamine, sodium silicate, or a combination thereof.

5. The concentrated multi-enzyme cleaning agent according to claim 1, characterized in that: The stabilizer is glycerol.

6. The concentrated multi-enzyme cleaning agent according to claim 1, characterized in that: The surfactant is: AEO-9 (fatty alcohol polyoxyethylene ether), decyl glucoside APG 0810, Brij 35 (lauryl alcohol polyoxyethylene ether), or a combination thereof.

7. The concentrated multi-enzyme cleaning agent according to claim 1, characterized in that: The cleaning agent includes a chelating agent.

8. The concentrated multi-enzyme cleaning agent according to claim 1, characterized in that: The cleaning agent contains the following components: protease 0.3-5 IU / mL; lipase 0.5-5 IU / mL; amylase 0.2-5 IU / mL; cellulase 0.2-5 IU / mL; triethanol and / or sodium silicate 3-10 w / w%; glycerol 20-30 w / w%; glycine and boric acid 1-2 mol / L; AEO-9 and / or decyl glucoside APG 0810 and / or Brij 35 0.05-1.0 mol / L; and the balance is deionized water.

9. The concentrated multi-enzyme cleaning agent according to claim 1, characterized in that: The protease is a protease from Bacillus, the lipase is a lipase from Candida rugosa, and / or the amylase is an amylase from Bacillus.

10. A method for cleaning an appliance, characterized in that: Includes steps: (a) providing a concentrated multi-enzyme cleaning agent as described in any one of claims 1 to 8, and diluting it 10 to 600 times to obtain a diluted multi-enzyme cleaning agent; (b) cleaning the utensil with the diluted multi-enzyme cleaning agent.

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