Keratin-based iodine-containing disinfectant and preparation method thereof

Abstract

The present application provides an iodine-containing disinfectant based on keratin and a preparation method thereof. The iodine-containing disinfectant comprises elemental iodine, keratin, a stabilizer and water. The preparation method of the iodine-containing disinfectant comprises: step 1): dissolving elemental iodine and a first stabilizer in water, then adding keratin, and then adjusting the pH to obtain a mixed solution A; step 2): adding a second stabilizer to the mixed solution A, mixing, and obtaining the iodine-containing disinfectant. In the iodine-containing disinfectant of the present application, by combining elemental iodine, a stabilizer and keratin, the iodine-containing disinfectant not only has a good bactericidal effect, but also can promote the growth of granulation tissue and accelerate wound healing.

Description

Technical Field

[0001] The present application belongs to the technical field of disinfectants, and specifically relates to an iodine-containing disinfectant and a preparation method thereof. Background Art

[0002] Iodine-containing disinfectants refer to disinfectants with iodine as the main bactericidal ingredient. They have low toxicity to tissues and are widely used in various aspects such as clinical practice, hygiene, daily life, livestock and poultry farming, aquaculture, public places and drinking water.

[0003] Keratin is a fibrous animal protein with connective and protective functions. It is a structural protein in ectoderm cells and is widely found in animal hair, feathers, and hooves. Keratin has excellent bioactivity and tissue compatibility, and can play an anti-inflammatory, hemostatic, nerve repair role, reduce wound exudate, and promote the regeneration, repair, and healing of wound tissue. It is biodegradable in the body, making it an excellent biomedical material.

[0004] However, when iodine-containing disinfectants are used in combination with keratin, there is a problem that the iodine content is significantly reduced. Summary of the Invention

[0005] In response to the problems existing in the prior art, the present application provides a keratin-based iodine-containing disinfectant and a preparation method thereof.

[0006] Specifically, this application involves the following:

[0007] 1. An iodine-containing disinfectant comprising elemental iodine, keratin, a stabilizer and water.

[0008] 2. The iodine-containing disinfectant according to claim 1, wherein

[0009] In the iodine-containing disinfectant, the mass fraction of elemental iodine is 0.01-1%, preferably 0.05-0.5%, more preferably 0.05-0.15%;

[0010] The mass fraction of the keratin is 0.05%-2%, preferably, the mass fraction of the keratin is 0.1%-1%;

[0011] The mass fraction of the stabilizer is 0.016%-1.6%. Preferably, the mass fraction of the stabilizer is 0.1%-0.8%.

[0012] 3. The iodine-containing disinfectant according to item 1, wherein the stabilizer comprises a first stabilizer and / or a second stabilizer; preferably, the first stabilizer is potassium iodide and the second stabilizer is potassium iodate;

[0013] More preferably, in the iodine-containing disinfectant, the mass fraction of the potassium iodide is 0.08%-8%, preferably, the mass fraction of the potassium iodide is 0.10%-0.40%;

[0014] The mass fraction of the potassium iodate is 0.08%-8%, preferably, the mass fraction of the potassium iodate is 0.10%-0.40%;

[0015] Preferably, the mass ratio of potassium iodate to potassium iodate is 1:1.

[0016] 4. The iodine-containing disinfectant according to item 1, wherein the iodine-containing disinfectant further comprises a buffer solution; preferably, the buffer solution is a mixed solution of NaH2PO4.2H2O and Na2HPO4.12H2O;

[0017] Preferably, the mass fraction of the buffer solution is 0.02%-2%, preferably 0.1-1%, more preferably 0.1-0.3%.

[0018] 5. The iodine-containing disinfectant according to item 4, wherein the mass fraction of NaH2PO4.2H2O in the iodine-containing disinfectant is 0.01%-1%, preferably, the mass fraction of NaH2PO4.2H2O is 0.05%-0.15%;

[0019] The mass fraction of the Na2HPO4.12H2O is 0.01%-1%. Preferably, the mass fraction of the NaH2PO4.12H2O is 0.05%-0.15%.

[0020] 6. The iodine-containing disinfectant according to item 1, further comprising a cosolvent for promoting the dissolution of the elemental iodine; preferably, the cosolvent is polyvinylpyrrolidone;

[0021] Preferably, in the iodine-containing disinfectant, the mass fraction of the co-solvent is 88%-91%.

[0022] 7. A method for preparing the iodine-containing disinfectant according to any one of items 1 to 6, comprising:

[0023] Step 1): dissolving elemental iodine and a first stabilizer in water, then adding keratin, and then adjusting the pH to obtain a mixed solution A;

[0024] Step 2): adding a second stabilizer to the mixed solution A and mixing to obtain the iodine-containing disinfectant.

[0025] 8. The preparation method according to item 7, wherein

[0026] The first stabilizer is potassium iodide; the second stabilizer is potassium iodate.

[0027] 9. The preparation method according to item 7, wherein:

[0028] In the step 1), the pH is adjusted to 3-7, preferably, the pH is adjusted to 4-6.

[0029] 10. The preparation method according to item 7, wherein:

[0030] In the iodine-containing disinfectant, the mass fraction of elemental iodine is 0.01-1%, preferably 0.05-0.5%, more preferably 0.05-0.15%;

[0031] The mass fraction of the keratin is 0.05%-2%, preferably 0.1%-1%;

[0032] The mass fraction of the potassium iodide is 0.08%-8%, preferably 0.10%-0.40%;

[0033] The mass fraction of the potassium iodate is 0.08%-8%, preferably 0.10%-0.40%.

[0034] 11. The preparation method according to item 7, wherein:

[0035] In the step 2), after the mixing, a buffer solution is added; preferably, the buffer solution is a mixed solution of NaH2PO4.2H2O and Na2HPO4.12H2O;

[0036] Preferably, in the iodine-containing disinfectant, the mass fraction of the NaH2PO4.2H2O is 0.01%-1%, preferably 0.05%-0.15%; the mass fraction of the Na2HPO4.12H2O is 0.01%-1%, preferably 0.05%-0.15%.

[0037] 12. The preparation method according to item 7, wherein:

[0038] In the step 1), the cosolvent, the elemental iodine and the first stabilizer are dissolved in water.

[0039] Beneficial effects:

[0040] 1) The combined use of iodine and keratin in the present application can synergistically enhance efficacy, not only sterilizing and disinfecting the wound surface, but also promoting the proliferation and migration of fibroblasts, thereby accelerating wound healing.

[0041] 2) The addition of stabilizers such as potassium iodide and potassium iodate in the present application can effectively maintain the stability of the iodine content, thereby making the iodine-containing disinfectant have a good bactericidal effect and can extend the shelf life of the product.

[0042] 3) The iodine-containing disinfectant of the present application has simple ingredients, thereby reducing production costs, and the process is simple and suitable for large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 The migration rates of fibroblasts when treated with the iodine-containing disinfectant of Example 1, the iodine-containing disinfectant of Example 20, and the iodine-containing disinfectant of Comparative Example 1 and compared with the blank control. DETAILED DESCRIPTION

[0044] The present application is further described below with reference to examples. It should be understood that the examples are only used to further illustrate and explain the present application and are not intended to limit the present application.

[0045] Unless otherwise defined, technical and scientific terms used in this specification have the same meaning as those commonly understood by those skilled in the art. Although methods and materials similar or identical to those described herein may be used in experiments or practical applications, the materials and methods are described herein below. In the event of a conflict, the present specification, including definitions, will prevail. In addition, the materials, methods, and examples are provided for illustrative purposes only and are not intended to be limiting. The present application is further described below with reference to specific examples, which are not intended to limit the scope of this application.

[0046] The present application provides an iodine-containing disinfectant, which includes elemental iodine, keratin, a stabilizer and water.

[0047] In the iodine-containing disinfectant of the present application, iodine has good bactericidal and disinfecting effects and can be used for sterilization, wound healing, etc.

[0048] The addition of keratin can accelerate wound healing, but on the other hand it will significantly reduce the iodine content. In this application, the addition of a stabilizer can not only exert the synergistic effect of keratin and iodine, but also minimize the reduction of iodine content.

[0049] In some embodiments of the present application, in the above-mentioned iodine-containing disinfectant, the mass fraction of elemental iodine is 0.01%-1%, preferably 0.05%-0.5%, for example, the mass fraction of elemental iodine is 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.30%, 0.31%, 0.32%, 0.33%, 0.34%, 0.36%, 0.37%, 0.38%, 0.39%, 0.40%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.50%, 0.51%, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%, 0.58%, 0.59%, 0.60%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.70%, 0.71%, 0.72%, 0.73%, 0.74%, 0.75%, 22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%.

[0050] In some embodiments of the present application, in the above-mentioned iodine-containing disinfectant, the mass fraction of keratin is 0.05%-2%, preferably 0.1%-1%, for example, the mass fraction of keratin is 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.15%, 0.20%, 0.25%, 0.30%, 0.35%, 0.40%, 0.45%, 0.50%, 0.55%, 0.60%, 0.65%, 0.70%, 0.75%, 0.80%, 0.85%, 0.90%, 0.95%, 1%, 1.05%, 1.1%, 1.15%, 1.20%, 1.25%, 1.30%, 1.35%, 1.40%, 1.45%, 1.50%, 1.55%, 1.60%, 1.65%, 1.70%, 1.75%, 1.80%, 1.85%, 1.90%, 1.95%, 2%. If the mass fraction of keratin is too low, it will not promote cell growth and accelerate healing; if the mass fraction of keratin is too high, the available iodine content will be reduced too much and the stability will be poor.

[0051] The elemental iodine can be any iodine that can be selected by a person skilled in the art without any limitation, and can be directly purchased or prepared by oneself.

[0052] The keratin can be any keratin selected by those skilled in the art without limitation. It can be purchased keratin or self-prepared keratin, such as the recombinant keratin prepared by the applicant. The keratin can also be epidermal keratin and / or cytokeratin.

[0053] In some embodiments of the present application, in the iodine-containing disinfectant, the stabilizer includes a first stabilizer and / or a second stabilizer; preferably, the first stabilizer is potassium iodide and the second stabilizer is potassium iodate. If the mass fraction of the stabilizer is too low, the available iodine content decreases too quickly; if the mass fraction of the stabilizer is too high, the iodine content exceeds the specified value, increasing costs.

[0054] In some embodiments of the present application, in the above-mentioned iodine-containing disinfectant, the mass fraction of potassium iodide is 0.08%-8%, preferably 0.10%-0.40%; for example, the mass fraction of potassium iodide is 0.08%, 0.10%, 0.12%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.21%, 0.23%, 0.24%, 0.25%, 0.30%, 0.35%, 0.40%, 0.45%, 0.50%, 0.55%, 0.60%, 0.65%, 0.70%, 0.75%, and 0.80%.

[0055] In some embodiments of the present application, in the above-mentioned iodine-containing disinfectant, the mass fraction of potassium iodate is 0.08%-8%, preferably 0.10%-0.40%; for example, the mass fraction of potassium iodate is 0.08%, 0.10%, 0.12%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.21%, 0.23%, 0.24%, 0.25%, 0.30%, 0.35%, 0.40%, 0.45%, 0.50%, 0.55%, 0.60%, 0.65%, 0.70%, 0.75%, and 0.80%.

[0056] In some embodiments of the present application, in the above-mentioned iodine-containing disinfectant, the iodine-containing disinfectant further includes a buffer solution, and the buffer solution can be any buffer solution suitable for the present application, such as phosphate buffer solution, glycine buffer solution, etc.

[0057] Preferably, the buffer solution is a mixed solution of NaH2PO4.2H2O and Na2HPO4.12H2O, wherein NaH2PO4.2H2O and Na2HPO4.12H2O are used simultaneously to form a buffer pair. In the iodine-containing disinfectant, before NaH2PO4.2H2O and Na2HPO4.12H2O are added, the solution is acidic; after NaH2PO4.2H2O and Na2HPO4.12H2O are added to the solution, the acidic ionization of dihydrogen phosphate is suppressed, and the alkaline hydrolysis of monohydrogen phosphate is strengthened, which alleviates the effect of the acidic substance on the acidity and alkalinity of the solution and improves the stability of the iodine-containing disinfectant. If the mass fraction of the buffer solution is too small, the solution is acidic, and the reaction of iodide ions and iodate ions increases. If the mass fraction of the buffer solution is too high, the problem of iodine precipitation is prone to occur.

[0058] In some embodiments of the present application, in the above-mentioned iodine-containing disinfectant, the mass fraction of NaH2PO4.2H2O is 0.01%-1%, preferably 0.05%-0.15%; for example, the mass fraction of NaH2PO4.2H2O is 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9 %, 0.95%, 1%; the mass fraction of Na2HPO4.12H2O is 0.01%-1%, preferably 0.05%-0.15%; for example, the mass fraction of Na2HPO4.12H2O is 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%.

[0059] The present application prepares an iodine-containing disinfectant with good stability and bactericidal effect by combining simple ingredients such as elemental iodine, keratin, stabilizer, buffer and water.

[0060] The present application provides a method for preparing the above-mentioned iodine-containing disinfectant, comprising: step 1): dissolving elemental iodine and a first stabilizer, potassium iodide, in water, then adding keratin, and then adjusting the pH to obtain a mixed solution A; step 2): adding a second stabilizer, potassium iodate, to the mixed solution A, and mixing to obtain the iodine-containing disinfectant.

[0061] In some embodiments of the present application, the pH is adjusted to 3-7, preferably, to 4-6. Since potassium iodate and potassium iodide react under acidic conditions, the stronger the acidity, the faster the reaction. After the mixture of povidone iodine and potassium iodide has a pH of about 2, directly adding potassium iodate will accelerate the reaction rate, resulting in reduced stability. Therefore, the present application reduces the reaction between potassium iodate and potassium iodide by first mixing povidone iodine and potassium iodide, then adjusting the pH, and then adding potassium iodate to avoid reducing the stability of the iodine-containing disinfectant.

[0062] In some embodiments of the present application, in step 2), after potassium iodate is added and mixed, a buffer is added; preferably, the buffer is a mixed solution of NaH2PO4.2H2O and Na2HPO4.12H2O.

[0063] In some embodiments of the present application, in step 1), a cosolvent, elemental iodine, and a first stabilizer, potassium iodide, are dissolved in water together; preferably, povidone iodine comprises the cosolvent and elemental iodine, that is, in step 1), povidone iodine and potassium iodide are dissolved in water together, wherein the mass fraction of elemental iodine in povidone iodine is 9%-12%.

[0064] Preliminary Example 1 Effect of Keratin on Available Iodine Content

[0065] 1. Experimental methods

[0066] Sample 1: Take 25 ml of iodine tincture (purchased from Chongqing Pukang) and add 5 ml of keratin stock solution (Haimers (Chongqing) Medical Biotechnology Co., Ltd.).

[0067] Sample 2: Take 25 ml of iodine tincture (purchased from Shandong Lierkang) and add 1 ml of keratin stock solution (Haimers (Chongqing) Medical Biotechnology Co., Ltd.).

[0068] Perform iodine content titration experiments on samples 1 and 2 to determine the effective iodine content. At the same time, perform iodine content titration experiments on two purchased iodine tinctures to determine the effective iodine content and compare the results.

[0069] 2. Experimental results

[0070]

[0071] 3. Results Analysis

[0072] Adding keratin stock solution to iodine tincture will reduce the effective iodine content in iodine tincture.

[0073] Effect of the amount and time of keratin addition on the available iodine content in Example 2

[0074] 1. Experimental methods

[0075] Sample 1: Take 25 ml of iodine tincture (purchased from Shandong Lierkang), add 1 ml of keratin stock solution (Haimers (Chongqing) Medical Biotechnology Co., Ltd.), and leave at room temperature for 1 day.

[0076] Sample 2: Take 75 ml of iodine tincture (purchased from Shandong Lierkang) and add 0.9 ml of keratin stock solution (Haimers (Chongqing) Medical Biotechnology Co., Ltd.).

[0077] Sample 3: Take 75 ml of iodine tincture (purchased from Shandong Lierkang) and add 0.09 ml of keratin stock solution (Haimers (Chongqing) Medical Biotechnology Co., Ltd.).

[0078] Samples 1, 2, and 3 were subjected to an iodine content titration experiment to determine the available iodine content, and the results were compared with the test results of Sample 2 in Preliminary Example 1.

[0079] 2. Experimental results

[0080]

[0081] 3. Results Analysis

[0082] When adding keratin stock solution to iodine tincture, the effective iodine content will decrease as the volume added increases. At the same time, the increase in the standing time after addition will further lead to a decrease in the effective iodine content.

[0083] Preliminary Example 3 54°C Accelerated Experiment to Observe the Effect of Keratin on Available Iodine Content

[0084] 1. Experimental methods

[0085] Take the remaining samples 2 and 3 of Example 2, divide them into 2 parts, and take 50 ml of iodine tincture (Shandong Lierkang) purchased from the market, divide them into 2 parts, and store them in a constant temperature incubator at 54°C. After 3 days and 5 days, take out one part each, perform iodine content titration experiment, determine the available iodine content, and compare the test results.

[0086] 2. Experimental results

[0087]

[0088] 3. Results Analysis

[0089] After being stored in a constant temperature incubator at 54℃ for 5 days, the available iodine content decreased, and the higher the keratin content, the greater the decrease.

[0090] Study on the stability of available iodine content of iodine-containing disinfectant in Example 4

[0091] 1. Experimental methods

[0092] Weigh 49.2019 g of povidone iodine (Boai Xinkaiyuan) and add purified water to prepare 500 ml of solution, which is recorded as sample 1.

[0093] Take 250 ml of sample 1, add 250 μl of keratin stock solution, mix well, and record it as sample 2.

[0094] Take 75 ml of sample 1 and 150 ml of sample 2, and package them into 25 ml portions. Divide them into 3 groups, with 1 portion of sample 1 and 2 portions of sample 2 as one group. Store one group in a constant temperature incubator at 37°C, and store another group in a constant temperature incubator at 54°C. Perform an iodine content titration experiment on the remaining group to determine the effective iodine content.

[0095] After 11 days, the samples stored in the 54°C constant temperature incubator were taken out and subjected to an iodine content titration experiment to determine the available iodine content; after 12 days, the samples stored in the 37°C constant temperature incubator were taken out and subjected to an iodine content titration experiment to determine the available iodine content.

[0096] 2. Experimental results

[0097]

[0098] 3. Results Analysis

[0099] The effective iodine content in povidone-iodine solution decreases rapidly and cannot be used as a formula. It is considered to add stabilizers in the future to increase the stability of the solution.

[0100] Preliminary Example 5 Screening of Stabilizer 1 with Better Stabilization Effect on Available Iodine Content

[0101] 1. Experimental methods

[0102] Take 40 ml of sample 1 and 80 ml of sample 2 in Example 4, place them in a 54°C constant temperature incubator for 14 days, take them out and let them cool to room temperature, and process them according to the method in the following table:

[0103]

[0104] An iodine content titration experiment was performed to determine the effective iodine content. Sample 2 was tested twice, and the remaining samples were placed in a constant temperature incubator at 60°C for 4 days. After being taken out and cooled, an iodine content titration experiment was performed to determine the effective iodine content.

[0105] 2. Experimental results

[0106]

[0107] 3. Results Analysis

[0108] After the sample was placed in a constant temperature incubator at 60°C for 4 days, the available iodine content decreased significantly and the formulation failed.

[0109] Preliminary Example 6 Screening of Stabilizer 2 with Better Stabilization Effect on Available Iodine Content

[0110] 1. Experimental methods

[0111] Prepare the sample by mixing the remaining povidone-iodine solution (containing only povidone-iodine) from the previous experiment. After mixing, the mass percentage is about 5%. Divide it into 3 portions of 60 ml each, for a total of 3 portions. Add different stabilizer formulas as shown in the table below:

[0112] sample Potassium iodide (mg) Potassium iodate (mg) EDTA (mg) Citric acid (mg) Adjust pH to Sample 1 62.49 73.50 60.87 / / Sample 2 63.45 72.91 60.62 / 5.0 Sample 3 600.99 / / 703.41 5.0

[0113] An iodine content titration experiment was performed to determine the effective iodine content. The remaining samples were placed in a constant temperature incubator at 60°C for 3 days. After being taken out and cooled, an iodine content titration experiment was performed to determine the effective iodine content.

[0114] 2. Experimental results

[0115]

[0116] 3. Results Analysis

[0117] Adding potassium iodate and potassium iodide at the same time will react to produce elemental iodine, increasing the effective iodine content. After the sample was placed in a constant temperature incubator at 60°C for 3 days, the effective iodine content decreased significantly, and the formula failed.

[0118] Preliminary Example 7 Screening of Stabilizer 3 with Better Stabilization Effect on Available Iodine Content

[0119] 1. Experimental methods

[0120] Prepare 8 groups of samples according to the table below, 100 ml each:

[0121]

[0122] Note: Adjust pH first, then add phosphate.

[0123] An iodine content titration experiment was performed to determine the available iodine content. The remaining samples were placed in a constant temperature incubator at 54°C for 2, 6, and 14 days. After being taken out and cooled, an iodine content titration experiment was performed to determine the available iodine content.

[0124] 2. Experimental results

[0125]

[0126]

[0127] 3. Results Analysis

[0128] Adding potassium iodate and potassium iodide at the same time will react to produce elemental iodine, increasing the available iodine content. In a phosphate buffer system, the available iodine content can be kept stable for a long time. Glycerol will destroy the balance after a certain period of time and accelerate the reduction of the available iodine content. Pyrrolidone has no obvious effect. After samples 5, 6, and 7 were placed in a constant temperature incubator at 54°C for 14 days, the available iodine content decreased by less than 10%. However, the available iodine content was too high and the formulation failed.

[0129] Preliminary Example 8 Screening of Stabilizer 4 with Better Stabilization Effect on Available Iodine Content

[0130] 1. Experimental methods

[0131] Reduce the amount of potassium iodide and potassium iodate to reduce the available iodine content to below 1%. Weigh 5.0156g of povidone iodine, 0.09947g of potassium iodide, and 0.05091g of potassium iodate, add 90ml of purified water, stir to dissolve, adjust the pH to 5.0, add 0.5g of NaH2PO4.2H2O and 0.1g of Na2HPO4.12H2O, and add purified water to 100ml.

[0132] An iodine content titration experiment was performed to determine the available iodine content. The remaining samples were placed in a constant temperature incubator at 54°C for 5 days. After being taken out and cooled, an iodine content titration experiment was performed to determine the available iodine content.

[0133] 2. Experimental results

[0134]

[0135] 3. Results Analysis

[0136] The available iodine content reached below 1%, but the stability was poor. After the sample was placed in a constant temperature incubator at 54°C for 5 days, the available iodine content decreased significantly and the formulation failed.

[0137] Preliminary Example 9 Screening of Stabilizer 5 with Better Stabilization Effect on Available Iodine Content

[0138] 1. Experimental methods

[0139] Prepare 3 groups of samples according to the table below, 100 ml each:

[0140]

[0141] Note: First, mix the three weighed groups of povidone-iodine, potassium iodide, and potassium iodate, and dissolve them in 36ml, 90ml, and 90ml of purified water, respectively. Then, mix the three groups of NaH2PO4.2H2O and Na2HPO4.12H2O, and dissolve them in 8ml, 20ml, and 20ml of purified water, respectively. Only half of this solution is added to the sample. After mixing the first group, add 60ml of purified water.

[0142] An iodine content titration experiment was performed to determine the available iodine content. The remaining samples were placed in a constant temperature incubator at 54°C for 2, 7, and 14 days. After being taken out and cooled, an iodine content titration experiment was performed to determine the available iodine content.

[0143] 2. Experimental results

[0144]

[0145] 3. Results Analysis

[0146] After samples 1 and 2 were placed in a constant temperature incubator at 54°C for 14 days, the available iodine content decreased by less than 10%, but the available iodine content was too high and the formulation failed.

[0147] Preliminary Example 10: Screening for a better stabilizer 6 for stabilizing the available iodine content

[0148] 1. Experimental methods

[0149] Prepare 2 groups of samples according to the table below, 90 ml each:

[0150]

[0151] Weigh 1.20072g of NaH2PO4.2H2O and 2.00058g of Na2HPO4.12H2O, add 40ml of purified water, and stir to dissolve. Add 10ml of the solution to each sample and mix.

[0152] An iodine content titration experiment was performed to determine the available iodine content. The remaining samples were placed in a constant temperature incubator at 54°C for 4, 7, and 14 days. After being taken out and cooled, an iodine content titration experiment was performed to determine the available iodine content.

[0153] 2. Experimental results

[0154]

[0155] 3. Results Analysis

[0156] The available iodine content was less than 1%, but after samples 1 and 2 were placed in a constant temperature incubator at 54°C for 14 days, the available iodine content decreased by more than 10%, indicating that the formulation failed.

[0157] Preliminary Example 11: Screening for a better stabilizer 7 for stabilizing the available iodine content

[0158] 1. Experimental methods

[0159] Weigh 6.00242 g of NaH2PO4.2H2O and 10.0022 g of Na2HPO4.12H2O, add 200 ml of purified water, stir to dissolve, then add 0.40190 g of potassium iodide and 0.40023 g of potassium iodate, stir to dissolve, and finally add 2.00068 g of povidone iodine, stir to dissolve.

[0160] An iodine content titration experiment was performed to determine the effective iodine content. The remaining samples were placed in a constant temperature incubator at 54°C and 37°C for 3 and 7 days respectively. After being taken out and cooled, an iodine content titration experiment was performed to determine the effective iodine content.

[0161] 2. Experimental results

[0162]

[0163] 3. Results Analysis

[0164] The available iodine content was less than 1%, but after the sample was placed in a constant temperature incubator at 54°C for 7 days, the available iodine content decreased by more than 10%, indicating that the formulation failed. At the same time, precipitation occurred in the sample.

[0165] Preliminary Example 12: Screening for a better stabilizer 8 for stabilizing the available iodine content

[0166] 1. Experimental methods

[0167] Solution preparation:

[0168] Solution 1: Weigh 2.00160 g of NaH2PO4.2H2O and 2.00062 g of Na2HPO4.12H2O, add 40 ml of purified water, and stir to dissolve.

[0169] Solution 2: Weigh 10.00237 g of povidone iodine and 2.00224 g of potassium iodide, add 180 ml of purified water, stir to dissolve, test the pH, which is 1.98, adjust the pH to 5.18 with 3 mol / L NaOH solution, then weigh 2.00290 g of potassium iodate and stir to dissolve.

[0170] Mix solutions 1 and 2 according to the table below to prepare the sample:

[0171]

[0172] In this preliminary embodiment, the solution preparation method is changed to first adding povidone iodine and potassium iodide, adjusting the pH, then adding potassium iodate, and finally adding NaH2PO4.2H2O and Na2HPO4.12H2O, and then adding purified water to dilute to the corresponding concentration.

[0173] An iodine titration experiment was performed to determine the available iodine content. The remaining samples were divided into two portions. One portion was placed in a 54°C constant temperature incubator for 3, 7, and 14 days. After being removed and cooled, an iodine titration experiment was performed to determine the available iodine content. The other portion was placed in a 37°C constant temperature incubator for 7, 14, 30, 60, and 90 days. After being removed and cooled, an iodine titration experiment was performed to determine the available iodine content.

[0174] 2. Experimental results

[0175] 54℃ results

[0176]

[0177] 37℃ results

[0178]

[0179] 3. Results Analysis

[0180] The available iodine content is less than 1%. After the sample is placed in a constant temperature incubator at 54°C for 14 days, the available iodine content decreases by less than 10%, indicating that the formula is initially successful.

[0181] Example

[0182] Example 1

[0183] Prepare the raw materials according to the mass ratio of each component in Example 1 in Table 1 below.

[0184] 4 / 5 of water is added to the mixture of NaH2PO4.2H2O and Na2HPO4.12H2O to obtain a mixed solution of NaH2PO4.2H2O and Na2HPO4.12H2O, i.e., a buffer solution.

[0185] The remaining 1 / 5 of water is added to a mixture of povidone-iodine (comprising a cosolvent polyvinylpyrrolidone and elemental iodine, wherein the mass fraction of iodine is 9-12%; purchased from Boai Xinkaiyuan) and potassium iodide, and the mixture is stirred and dissolved. Keratin is then added, wherein the keratin is the recombinant keratin modified in CN 115531239 A. 3 mol / L NaOH solution is then used to adjust the pH to 5.00, potassium iodate is added, and the mixture is stirred and dissolved. Finally, a buffer solution is added and the mixture is stirred and mixed to obtain an iodine-containing disinfectant. The iodine-containing disinfectant comprises: 1% by mass of iodine, 0.5% by mass of keratin, 0.2% by mass of potassium iodide, 0.2% by mass of potassium iodate, 0.1% by mass of NaH2PO4.2H2O, 0.1% by mass of Na2HPO4.12H2O, and 97.9% by mass of water.

[0186] After obtaining the iodine-containing disinfectant, the iodine-containing disinfectant was stored at 54° C. for 7 days and 14 days, respectively. After cooling, an iodine content titration experiment was performed to determine the effective iodine content. The result data are listed in Table 1 below.

[0187] Example 2

[0188] The only difference between Example 2 and Example 1 is that the mass fraction of keratin is 0.2%, and the rest is the same as Example 1.

[0189] Example 3

[0190] The only difference between Example 3 and Example 1 is that the mass fraction of keratin is 0.3%, and the rest is the same as Example 1.

[0191] Example 4

[0192] The only difference between Example 4 and Example 1 is that the mass fraction of keratin is 0.7%, and the rest is the same as Example 1.

[0193] Example 5

[0194] The only difference between Example 5 and Example 1 is that the mass fraction of keratin is 1.0%, and the rest is the same as Example 1.

[0195] Example 6

[0196] The only difference between Example 6 and Example 1 is that the mass fraction of keratin is 2.0%, and the rest is the same as Example 1.

[0197] Example 7

[0198] The difference between Example 7 and Example 1 is only that the mass fraction of potassium iodide is 0.15%, and the rest is the same as Example 1.

[0199] Example 8

[0200] The only difference between Example 8 and Example 1 is that the mass fraction of potassium iodide is 0.25%, and the rest is the same as Example 1.

[0201] Example 9

[0202] The only difference between Example 9 and Example 1 is that the mass fraction of potassium iodate is 0.15%, and the rest is the same as Example 1.

[0203] Example 10

[0204] The difference between Example 10 and Example 1 is that the mass fraction of potassium iodate is 0.25%, and the rest is the same as Example 1.

[0205] Example 11

[0206] The difference between Example 11 and Example 1 is that the mass fraction of potassium iodide is 0.1%, the mass fraction of potassium iodate is 0.1%, and the rest is the same as Example 1.

[0207] Example 12

[0208] The difference between Example 12 and Example 1 is only that the mass fraction of potassium iodide is 0.3%, the mass fraction of potassium iodate is 0.3%, and the rest is the same as Example 1.

[0209] Example 13

[0210] The only difference between Example 13 and Example 1 is that 3 mol / L NaOH solution is used to adjust the Ph to 3, and the rest is the same as Example 1.

[0211] Example 14

[0212] The only difference between Example 14 and Example 1 is that 3 mol / L NaOH solution is used to adjust the Ph to 4, and the rest is the same as Example 1.

[0213] Example 15

[0214] The only difference between Example 15 and Example 1 is that 3 mol / L NaOH solution is used to adjust the Ph to 6, and the rest is the same as Example 1.

[0215] Example 16

[0216] The only difference between Example 16 and Example 1 is that 3 mol / L NaOH solution is used to adjust the Ph to 7, and the rest is the same as Example 1.

[0217] Example 17

[0218] The only difference between Example 15 and Example 1 is that the mass fraction of NaH2PO4.2H2O is 0.5%, the mass fraction of Na2HPO4.12H2O is 0.5%, and the rest is the same as Example 1.

[0219] Example 18

[0220] The only difference between Example 18 and Example 1 is that the mass fraction of NaH2PO4.2H2O is 1.0%, the mass fraction of Na2HPO4.12H2O is 1.0%, and the rest is the same as Example 1; after the obtained iodine-containing disinfectant is stored at 54°C for 7 days, obvious iodine elemental precipitation occurs and settles to the bottom.

[0221] Example 19

[0222] The only difference between Example 19 and Example 1 is that the mass fraction of NaH2PO4.2H2O is 2.0%, the mass fraction of Na2HPO4.12H2O is 2.0%, and the rest is the same as Example 1; after the obtained iodine-containing disinfectant is stored at 54°C for 7 days, obvious iodine elemental precipitation occurs and settles to the bottom.

[0223] Example 20

[0224] The only difference between Example 20 and Example 1 is that the keratin in this example is commercially available keratin, which is wool-extracted keratin purchased from Shanghai Huilang Chemical Co., Ltd., with the trade name: FK Protect Plus and the batch number: 21C10B188.

[0225] Comparative Example 1

[0226] The only difference between Comparative Example 1 and Example 1 is that the mass fraction of keratin is 0%, and the rest is the same as Example 1.

[0227] Table 1

[0228]

[0229]

[0230] PS: In the above-described embodiment and comparative example, when the component (excluding water) content in the iodine-containing disinfectant changes, the content of water changes accordingly so that the content of the total components is 100%. In Table 1, the 0-day available iodine content is the content of iodine after the stabilizer potassium iodide and potassium iodate and povidone-iodine act together.

[0231] The bactericidal effect of the iodine-containing disinfectant obtained in Example 1 was verified by referring to the laboratory bactericidal effect test method of disinfectants in GB / T 38502-2020. The results showed that after adding the iodine-containing disinfectant of Example 1 of the present application, no colonies grew, indicating a good disinfection effect.

[0232] Perform cell scratch assay

[0233] L929 fibroblasts were cultured and digested when they reached 95% confluence. Cells were collected by centrifugation at 1000 rpm for 5 minutes and suspended in complete serum-containing DMEM medium for cell counting. Cells were seeded into 6-well plates at a concentration of 2 × 105 cells / well in 1.5 mL of culture medium and incubated at 37°C in a 5% CO2 incubator until the cells reached 90% confluence on the next day. After the cells adhered, a 200 μl pipette tip was used to create a blank area by drawing a cross vertically against the bottom of the 6-well plate. Floating cells were washed with PBS buffer and then DMEM basal medium was added.

[0234] Comparison was made between a blank control (PBS-cultured cells), cells treated with an iodine-containing disinfectant in Example 1 (containing 0.5% recombinant keratin), Example 20 (containing 0.5% commercially available keratin), and Comparative Example 1 (a control group without keratin). Images were taken of the same area using a microscope at 24 and 48 hours of treatment. The area of cell migration was analyzed using ImageJ software. The cell healing rate was calculated using the formula to represent the cell migration rate.

[0235] Healing rate = [(A0-Ax) / A0] × 100

[0236] Where A0 represents the area of the initial scratch area of the cells, and Ax represents the area of the scratch area of the cells at different time points after treatment.

[0237] The repair activity of disinfection products containing recombinant keratin was tested by cell scratch test. The results (see attached) Figure 1 (as shown in Figure 2) shows that when 24h, the fibroblasts treated with the iodine-containing disinfectant (Example 20) containing 0.5% commercially available keratin had a migration rate greater than 40%, and the migration rate of the cells treated with the iodine-containing disinfectant (Example 1) containing recombinant keratin reached nearly 60%, while the cells cultured in culture medium and the cells treated with the iodine-containing disinfectant (Comparative Example 1) containing no keratin had migration rates of 22.13% and 27.05% respectively, and the cell migration speed was significantly lower than the keratin group of Example 1. During the 48th hour, the effect was even more obvious. The migration rate of the cells treated with the iodine-containing disinfectant (Example 1) containing 0.5% recombinant keratin reached more than 90%, the migration rate of the cells treated with the iodine-containing disinfectant (Example 20) containing commercially available keratin was less than 60%, and the other two groups (cells cultured in culture medium and cells treated with the iodine-containing disinfectant (Comparative Example 1) containing no keratin) only had about 40% migration rate. The cell scratch results show that the cells cultured with the iodine-containing disinfectant (Example 1) containing 0.5% recombinant keratin had the best cell migration rate.

Claims

1. An iodine-containing disinfectant, characterized in that, Comprising elemental iodine, keratin, a stabilizer, a buffer and water; wherein the mass fraction of the elemental iodine is 1%; The mass fraction of the keratin is 0.2%-1%; The stabilizer includes a first stabilizer and a second stabilizer; the first stabilizer is potassium iodide, and the second stabilizer is potassium iodate; the mass fraction of the potassium iodide is 0.10%-0.30%; the mass fraction of the potassium iodate is 0.10%-0.30%; and the mass ratio of the potassium iodate to the potassium iodide is 1:1; The buffer solution is a mixed solution of NaH2PO4.2H2O and Na2HPO4.12H2O, the mass fraction of the buffer solution is 0.2%, the mass fraction of the NaH2PO4.2H2O is 0.1%, and the mass fraction of the NaH2PO4.12H2O is 0.1%; The pH of the iodine-containing disinfectant is 5.

2. iodine-containing disinfectant according to claim 1, is characterized in that, In the iodine-containing disinfectant, the mass fraction of the potassium iodide is 0.2%; the mass fraction of the potassium iodate is 0.20%.

3. iodine-containing disinfectant according to claim 1, is characterized in that, In the iodine-containing disinfectant, the mass fraction of keratin is 0.2%-0.7%.

4. iodine-containing disinfectant according to claim 1, is characterized in that, The iodine-containing disinfectant also includes polyvinyl pyrrolidone, which is used to promote the dissolution of the elemental iodine.

5. A method for preparing an iodine-containing disinfectant according to any one of claims 1 to 4, characterized in that: include: Step 1): dissolving elemental iodine and potassium iodide in water, then adding keratin, and adjusting the pH to 5 to obtain a mixed solution A; Step 2): adding potassium iodate to the mixed solution A, mixing, then adding a buffer solution, and mixing to obtain the iodine-containing disinfectant; Wherein, in the iodine-containing disinfectant, the mass fraction of the elemental iodine is 1%; The mass fraction of the keratin is 0.2%-1%; The mass fraction of the potassium iodide is 0.10%-0.30%; the mass fraction of the potassium iodate is 0.10%-0.30%; the mass ratio of the potassium iodate to the potassium iodide is 1:1; The buffer solution is a mixed solution of NaH2PO4.2H2O and Na2HPO4.12H2O, the mass fraction of the buffer solution is 0.2%, the mass fraction of the NaH2PO4.2H2O is 0.1%, and the mass fraction of the NaH2PO4.12H2O is 0.1%.

6. The preparation method according to claim 5, characterized in that In the iodine-containing disinfectant, the mass fraction of the potassium iodide is 0.2%; the mass fraction of the potassium iodate is 0.20%.

7. The preparation method according to claim 5, characterized in that The mass fraction of the keratin is 0.2-0.7%.

8. The preparation method according to claim 5, characterized in that In the step 1), polyvinyl pyrrolidone, elemental iodine and potassium iodide are dissolved in water.

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