CN114931138A

CN114931138A - Sterilization disinfectant and preparation method thereof

Info

Publication number: CN114931138A
Application number: CN202111190850.9A
Authority: CN
Inventors: 倪卓
Original assignee: Shenzhen Huabao Biomaterial Technology Co ltd
Current assignee: Shenzhen Huabao Biomaterial Technology Co ltd
Priority date: 2021-10-13
Filing date: 2021-10-13
Publication date: 2022-08-23
Anticipated expiration: 2041-10-13
Also published as: CN114931138B

Abstract

The invention provides a sterilizing disinfectant and a preparation method thereof, wherein the effective components of the sterilizing disinfectant are in a microcapsule form, and the capsule core contains two chemical substances, wherein one chemical substance is polyhexamethylene guanidine hydrochloride or polyhexamethylene guanidine phosphate, and the other chemical substance is benzalkonium chloride or benzalkonium bromide. The microcapsule wall structure has the characteristic of chemical water-soluble gel, can be compatible with soap, anionic surfactant and the like, and overcomes the defect that the application of the existing polyhexamethylene guanidine salt in washing products is limited because most of the washing product formulas in the market contain the anionic surfactant. On the other hand, by wrapping polyhexamethylene guanidine hydrochloride (phosphate) and benzalkonium chloride (bromide) aqueous solution with a disinfection effect in a capsule wall structure with certain mechanical properties, the capsule wall structure of the microcapsule is destroyed when in use, and the polyhexamethylene guanidine hydrochloride (phosphate) and benzalkonium chloride (bromide) aqueous solution are released for sterilization and disinfection.

Description

Bactericidal disinfectant and preparation method thereof

Technical Field

The invention relates to the technical field of medical treatment, in particular to a sterilizing disinfectant and a preparation method thereof.

Background

The types of disinfectants commonly used in the prior art are as follows:

1. oxidizing agents

The povidone iodine is commonly called iodophor and is a complex compound of polyvinylpyrrolidone and iodine. Iodophor is a dark red solution, odorless, tasteless, and free of insoluble substances, and can be mixed with water and ethanol. The iodophor can permeate and oxidize to swell the outer membrane layer, shell layer and core of spore, destroy the permeability of cell membrane, and iodinate bioactive substance molecules contacting with iodophor to make them lose activity, thereby killing bacteria. The iodophor has strong bactericidal power and has strong killing effect on bacteria, fungi, viruses, spores and the like. It kills bacterial propagules very rapidly, but killing spores generally requires higher concentrations and longer times.

② the hydrogen peroxide for hydrogen peroxide disinfection has 3 percent of use concentration and is mainly used for wound disinfection. The hydrogen peroxide forms free hydroxyl with strong oxidizing ability when contacting with bacteria, destroys the basic molecular structure of protein and destroys bacteria thallus, thereby playing the role of bacteriostasis and sterilization. After killing bacteria, green decomposition products of water and oxygen are produced, and environmental pollution is avoided. Therefore, hydrogen peroxide is an ideal disinfectant.

2. Surfactants and the like

Benzalkonium chloride (bromide) is a quaternary ammonium salt cationic surfactant and has strong bactericidal performance in various bactericidal surfactants. The benzalkonium chloride (bromide) solution can adsorb bacteria with negative charges, destroy cell membranes of the bacteria, enable enzymes, coenzymes and metabolic intermediates in bacteria to escape, and denature and precipitate mycoprotein. Has strong bacteriostasis and sterilization effects on a plurality of spore pathogenic bacteria, gram-positive bacteria and mould fungi, but has weaker reaction on gram-negative bacteria and enteroviruses and has no effect on tubercle bacillus. The benzalkonium chloride (bromide) has low bactericidal concentration, less side reaction, no irritation and difficult volatilization. However, when benzalkonium chloride (bromide) is used, attention should be paid to the effect of hydrogen peroxide and soapy water on the disinfection effect of the disinfectant. Free hydroxyl formed by hydrogen peroxide has negative charges, the effective part of the benzalkonium chloride (bromide) is a cationic group which has positive charges, and the positive charges and the negative charges are mutually attracted and combined, so that the sterilization effect is obviously reduced. Soap is an anionic detergent, and if the soap meets benzalkonium chloride (bromide), the bactericidal capacity of the disinfectant is weakened or even lost due to the attraction of positive and negative charges.

3. Alcohols

Ethanol belongs to an alcohol intermediate-effect disinfectant, and absorbs the moisture of bacterial protein to ensure that the bacterial protein is dehydrated, denatured and solidified, thereby killing bacteria. An aqueous solution with a volume fraction of about 75% is the optimum concentration for the use of ethanol as a disinfectant. The reasons are as follows: (1) the ethanol molecule enters the peptide chain link of the bacterial protein molecule, so that the protein is denatured and precipitated, and the effect is stronger when the volume fraction of the ethanol is 70%. (2) And (3) breaking the bacterial cell wall: the ethanol has strong osmosis, and 60-85% of ethanol can easily permeate into the thallus, so that the bacterial cells are destroyed and dissolved. (3) And (3) destroying a microbial enzyme system: ethanol inhibits the normal metabolism and inhibits the growth and reproduction of bacteria by inhibiting the bacterial enzyme system, particularly dehydrogenase and oxidase. If high-concentration ethanol is used, bacterial protein is dehydrated too quickly, so that the protein on the surface of the bacteria is firstly denatured and solidified to form a firm coating, and the ethanol cannot well penetrate into the bacteria instead, so that the sterilization capability of the bacteria is influenced. The osmotic pressure of 75% ethanol is similar to that of bacteria, and the ethanol can gradually permeate into thalli before the protein on the surface of the bacteria is not denatured, so that all the protein of the bacteria is dehydrated, denatured and coagulated, and finally the bacteria are killed. Ethanol concentrations below 75% may also affect bactericidal efficacy due to reduced permeability. Ethanol is used for disinfection, but is nontoxic to human body, but individual people are allergic to ethanol and can cause rash and erythema after contact. Frequent use of ethanol for hand washing and disinfection can cause dry and rough skin, and the problem can be solved by adding humectants such as glycerol, ethylene glycol and the like into the ethanol disinfectant.

4. Organic small molecules

Triclosan, commonly known as triclosan, is a widely used broad-spectrum antimicrobial agent in the scientific name of 2,4,4 '-trichloro-2' -hydroxydiphenyl ether, and is used in household daily necessities and personal hygiene products because of its high sterilization efficiency, no irritation to the skin and good skin compatibility. Such broad-spectrum antimicrobial agents are also widely used in medical device disinfectants, disinfection of textiles before shipment, toys and construction materials. Triclosan is a white or off-white crystalline powder with strong surface activity, phenol odor, solubility in organic solvents, and high stability to strong acids, bases and heat. The basic characteristics of triclosan are shown in the following six aspects. The product has high purity and is not easy to be polluted by skin and products. ② slightly soluble in water and easily soluble in various organic solvents and surfactants. And thirdly, the storage is easy, and the high stability to strong acid, strong base and heat is realized. The material can not be rapidly decomposed when stored at high temperature; the decomposition is only slight under the irradiation of ultraviolet rays for a long time; the active substance is only decomposed by 2% when heated for 14h at 200 ℃. And fourthly, the bactericide has the functions of inhibiting and killing gram-positive bacteria, saccharomycetes, fungi, viruses and the like. Fifth, no irritation to skin. Sixthly, the antibacterial agent is also effective to drug-resistant bacteria and non-drug-resistant bacteria.

5. Polymers of the class

The polymer disinfectant is a new generation of bactericide, has excellent bactericidal activity, and has the following advantages: high efficiency, safety, no toxic and side effect, and stability to light and heat; does not generate bacterial drug resistance and is effective for a long time; colorless, tasteless and non-volatile; heavy metal and phenolic substances are not contained; no corrosion to various treated surfaces; is environment-friendly.

The common polymer disinfectant is polyhexamethylene guanidine disinfectant. Polyhexamethylene guanidine disinfectants are classified into biguanide salts and monoguanidine salts. The commonly used biguanide salt is polyhexamethylene biguanide hydrochloride, which is generally sold as an aqueous solution with the mass fraction of 20 percent and can be used alone or in combination with other types of bactericides. The pH value of the system is 4-10, the odor and the foam are reduced to a large extent compared with quaternary ammonium salt disinfectants, and the active ingredients are stable to heat and do not volatilize. The monoguanidine salts are: 1) polyhexamethyleneguanidine hydrochloride. No special smell, easy to dissolve in water; the water solution is colorless to light yellow, tasteless and has weak corrosion to metals such as copper, stainless steel, carbon steel and the like; broad-spectrum sterilization and good stability. 2) Polyhexamethylene guanidine stearate. The derivative of the mono-guanidine salt disinfectant has good stability, and the antibacterial activity of the derivative is still good after the derivative is heated for 15min at the high temperature of 280 ℃. 3) Polyhexamethylene guanidine propionate. Has excellent sterilizing effect, low toxicity and no corrosion to metal. Generally made into powder, and has wide application in the fields of plastics, daily chemicals, textiles, water treatment and the like. 4) Polyhexamethylene guanidine phosphate. The compound containing phosphate radical has irritation to skin mucous membrane and eyes when being used as disinfectant, has high toxicity, and simultaneously contains phosphorus as growth substance of algae, so that water tends to eutrophication, thus destroying environment. Polyhexamethylene guanidine phosphate is not widely used at present and is also used internationally with great care. 5) Polyhexamethylene guanidine sulfate. A white powder is odorless, easily soluble in water, and substantially non-corrosive to metals, and has a decomposition temperature higher than 400 deg.C, no bleaching phenomenon on the treated surface, and no deterioration for two years. The bactericidal performance of the polyhexamethylene biguanide salt is similar to that of the biguanide salt, the synthesis cost of the monoguanidine salt is low, the biguanide salt is milder, the safety is higher, and the bactericidal effect of the hydrochloride is better than that of other salts.

The sterilization mechanism of the polyhexamethylene guanidine salt disinfectant mainly comprises the following steps: 1) the guanidino in the polyhexamethylene guanidine salt has high activity, and the polymer is electropositive. Because various bacteria and viruses are generally electronegative, the polyhexamethylene guanidine salt is easily adsorbed, so that the bacteria and the viruses cannot split and propagate and lose activity; 2) the structure of the cell membrane is collapsed to form a transmembrane pore, the cell membrane is broken to destroy the energy metabolism of the microorganism, so that the activity of the bacterial virus is lost; 3) the polymer can form a layer of film to seal the breathing channel of the microorganism, so that the microorganism is suffocated. The sterilization mechanism is irrelevant to the form and the type of the microorganism, the effect of the microorganism is not influenced even if the microorganism is changed or mutated, and the microorganism does not generate the drug resistance to the polyhexamethylene guanidine salt.

The existing polyhexamethylene guanidine salt disinfectant has cationic property and is difficult to be compatible with soap, anionic surfactant and the like, and most of washing product formulas in the market contain the anionic surfactant, so that the application of the polyhexamethylene guanidine salt in washing products is limited.

Disclosure of Invention

The invention aims to solve the defects of the prior art and provides a sterilizing disinfectant and a preparation method thereof.

In order to achieve the purpose, the invention adopts the following technical scheme: a germicidal sterilant, characterized by: the microcapsule comprises a microcapsule with a core material of polyhexamethylene guanidine hydrochloride (phosphate) and a benzalkonium chloride (bromide) aqueous solution, wherein the wall material of the microcapsule is a polymer formed by crosslinking citric acid/genipin and gelatin/chitosan, and the chemical structural formula of the polymer is shown as the formula (I):

in the formula (I), R1-R9 are selected from one of residues of eighteen different amino acids of glycine, alanine, serine, aspartic acid, glutamic amino acid, proline, arginine, histidine, tyrosine, cystine, leucine, threonine, methionine, valine, phenylalanine, tryptophan, glutamic acid and lysine.

Preferably, in said formula (I):

r5 and R6 are selected from one of eighteen different amino acid residues of glycine, alanine, serine, aspartic acid, glutamic amino acid, proline, arginine, histidine, tyrosine, cystine, leucine, threonine, methionine, valine, phenylalanine, tryptophan, glutamic acid and lysine;

r3, R7 are residues of lysine or arginine;

r2, R4 and R8 are residues of aspartic acid or glutamic acid amino;

r1, R9 are residues of serine, threonine or tyrosine.

The invention also provides a preparation method of the sterilizing disinfectant, which comprises the following steps:

(1) mixing glycerol, propylene glycol, water-soluble cherry essence and deionized water uniformly;

(2) the preparation method of the microcapsule with the core material of polyhexamethylene guanidine hydrochloride (phosphate) and benzalkonium chloride (bromide) aqueous solution comprises the following steps:

a. dissolving gelatin in an acetic acid aqueous solution to obtain a gelatin acetic acid solution;

b. adding chitosan into gelatin acetic acid solution, stirring to dissolve chitosan to obtain gelatin/chitosan mixed solution, adding polyhexamethylene guanidine hydrochloride (phosphate) and benzalkonium chloride (bromide) aqueous solution, and adjusting pH to 5.8-6.2;

c. adding surfactant soybean phospholipid into vegetable oil, heating, and stirring;

d. c, adding the gelatin/chitosan mixed solution prepared in the step b into the vegetable oil obtained in the step c, heating and emulsifying, turning off the heating after the emulsification is finished, and naturally cooling to room temperature;

e. d, adding genipin into the solution system obtained after the step d for crosslinking reaction, and fully reacting;

f. adding citric acid and glacial acetic acid into the reaction system after the step e, adjusting the pH to 2-3, and adding N ₂ Fully reacting under protection to obtain microcapsules of which the capsule core substances are polyhexamethylene guanidine hydrochloride (phosphate) and aqueous solution of benzalkonium chloride (bromide);

(3) and (2) adding microcapsules with core substances of polyhexamethylene guanidine hydrochloride (phosphate) and benzalkonium chloride (bromide) aqueous solution into the system in the step (1) to prepare the sterilizing disinfectant.

Preferably, the mixture ratio of each component in the step (1) is as follows according to parts by weight: 5-10 parts of glycerol, 5-10 parts of propylene glycol and 0.2-0.5 part of water-soluble cherry essence; the ratio of the total mass of the components in the step (1) to the total mass of the components in the step (2) is 1: 1.

Preferably, step f is followed by: and f, standing the reaction system after the step f, pouring out an upper oil phase, centrifuging, and separating out the oil phase to obtain the microcapsule with the core substances of polyhexamethylene guanidine hydrochloride (phosphate) and benzalkonium chloride (bromide) aqueous solution.

Preferably, in step b, the pH is preferably adjusted to 6.

Preferably, the specific steps of step e are: and d, adding 0.5% genipin aqueous solution into the solution system obtained in the step d, reacting for 3 hours at room temperature, heating to 35 ℃, reacting for 15 hours again, and fully reacting.

Preferably, the specific steps of step f are: and e, adding a 1% citric acid aqueous solution into the reaction system in the step e, adding glacial acetic acid, adjusting the pH value to 2-3, raising the reaction temperature to 40 ℃ under the protection of N2, reacting for 8 hours, and cooling to room temperature to obtain the microcapsule with the capsule core substances of polyhexamethylene guanidine hydrochloride (phosphate) and benzalkonium chloride (bromide) aqueous solution.

Preferably, the vegetable oil of step c is one or more of corn oil, olive oil, soybean oil and peanut oil.

Preferably, the ratio of each component is as follows according to parts by weight: 45-55 parts of gelatin, 3-7 parts of chitosan, 350 parts of 1.0% acetic acid solution 250-ion, 40-60 parts of soybean lecithin, 30-40 parts of 0.5% genipin aqueous solution, 5-15 parts of 1.0% citric acid solution and 3000 parts of vegetable oil 2500-ion.

Further preferably, the composition comprises 50 parts by weight of gelatin, 5 parts by weight of chitosan, 300 parts by weight of 1.0% acetic acid solution, 50 parts by weight of soybean lecithin, 50 parts by weight of 0.5% genipin aqueous solution, 10 parts by weight of 1.0% citric acid solution and 2700 parts by weight of vegetable oil.

Preferably, the active ingredients of the sterilizing disinfectant are in a microcapsule form, the capsule core material comprises two chemical substances, wherein one chemical substance is polyhexamethylene guanidine hydrochloride or polyhexamethylene guanidine phosphate, the other chemical substance is benzalkonium chloride or benzalkonium bromide, and the mass ratio of the two chemical substances is 3:1-5:1, and further preferably is 4: 1.

Preferably, the capsule core material is 8-12 parts of germicidal disinfectant and 350-400 parts of water, and further preferably, the capsule core is 10 parts of germicidal disinfectant and 380 parts of water.

Preferably, the mass ratio of gelatin to chitosan is 8: 1 to 12:1, the volume ratio of the water phase to the oil phase is 1:3 to 1: 5; further preferably, the mass ratio of gelatin to chitosan is selected to be 10:1, and the volume ratio of water phase to oil phase is selected to be 1: 4.

Compared with the prior art, the sterilizing disinfectant has the beneficial effects that the capsule core materials of the sterilizing disinfectant are two chemical substances, wherein one chemical substance is polyhexamethylene guanidine hydrochloride or polyhexamethylene guanidine phosphate, and the other chemical substance is benzalkonium chloride or benzalkonium bromide. The microcapsule wall structure has the characteristic of chemical water-soluble gel, can be compatible with soap, anionic surfactant and the like, and overcomes the defect that the application of the existing polyhexamethylene guanidine salt in washing products is limited because most of the washing product formulas in the market contain the anionic surfactant. On the other hand, by wrapping the polyhexamethylene guanidine hydrochloride (phosphate) and the aqueous solution of the benzalkonium chloride (bromide) in a capsule wall structure with certain mechanical properties, the capsule wall structure of the microcapsule is destroyed when in use, and the aqueous solution of the polyhexamethylene guanidine hydrochloride (phosphate) and the benzalkonium chloride (bromide) is released for sterilization and disinfection.

Drawings

FIG. 1 is a graph of isoelectric points of gelatin, chitosan, and chitosan/gelatin of example 1 at different mass ratios;

FIG. 2 is a thermogravimetric plot of gelatin, chitosan, gelatin/chitosan complex, genipin cross-linked gelatin/chitosan, and citric acid secondary cross-linked gelatin/chitosan.

FIG. 3 is a comparative microencapsulation microscope photograph showing that the core material of example 2 is an aqueous solution of polyhexamethylene guanidine phosphate and benzalkonium chloride.

Figure 4 is a microscopic image of the microcapsules after shearing in example 4.

FIG. 5 is a graph showing the bactericidal effect of the disinfectant prepared in example 2 on Staphylococcus aureus.

FIG. 6 is a graph showing the bactericidal effect of the bactericidal disinfectant prepared in example 2 on Escherichia coli.

Detailed Description

The technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention.

EXAMPLE 1 measurement of isoelectric Point of gelatin, Chitosan, gelatin/Chitosan composite

1.1 isoelectric Point of gelatin

0.5g of gelatin is weighed, 100ml of deionized water is added, and the mixture is stirred at a water bath temperature of 50 ℃ until the gelatin is completely dissolved, so that a 0.5% gelatin solution is obtained. The pH was adjusted using 0.001mol/L HCl solution and 0.001mol/L NaOH solution, and the conductivity of the gelatin solution at different pH values was recorded using a pH meter and conductivity meter.

1.2 isoelectric points of Chitosan

0.5g of chitosan is weighed and added into 100ml of 0.01mol/L HCl solution, and the mixture is stirred at room temperature until the chitosan is completely dissolved, so that 0.5 percent chitosan solution is obtained. The pH of the solution was adjusted using 0.001mol/L NaOH solution and the conductivity of the gelatin solution at different pH values was recorded using a pH meter and conductivity meter.

1.3 isoelectric point of Chitosan/gelatin composite

100mL of 1.0% gelatin solution was placed in a beaker and magnetically stirred with a water bath temperature of 50 ℃. Adding a certain volume of 1.0% chitosan solution into the gelatin solution, and stirring for 1h to obtain a gelatin/chitosan uniform mixed solution. The volume ratio of the chitosan to the gelatin is respectively as follows: 4:100, 10:100, 20: 100. 50:100, 75:100, 100: 100. The pH of the mixed solution was changed using 1.0% HCl solution and 0.1mol/L NaOH solution, and the conductivity of the gelatin/chitosan solution at various pH values was measured using a pH meter and a conductivity meter.

The isoelectric points of the gelatin and the chitosan obtained by the method and the isoelectric points of the chitosan/gelatin with different mass ratios are shown in Table 1, and the isoelectric point curve chart of the gelatin, the chitosan and the chitosan/gelatin with different mass ratios is shown in FIG. 1.

TABLE 1 isoelectric points of gelatin, chitosan and chitosan/gelatin in different mass ratios

EXAMPLE 2 preparation of germicidal sterilant

(1) Weighing 8.00g of glycerol, 8.00g of propylene glycol, 0.30g of water-soluble cherry essence and 83.70g of deionized water, and stirring for 10-20min for uniform mixing;

(2) the preparation method of the microcapsule with the core material of polyhexamethylene guanidine phosphate and benzalkonium chloride aqueous solution comprises the following steps:

a. weighing 5.00g of gelatin in a beaker, dissolving 30mL of 1.0% acetic acid aqueous solution at the water bath temperature of 37 ℃;

b. adding 0.50g of chitosan into the gelatin solution when the gelatin is completely dissolved, stirring for dissolving to obtain a uniform gelatin/chitosan mixed solution, adding polyhexamethylene guanidine phosphate and benzalkonium chloride, and adjusting the pH value to 6 by using a 5.0% ammonia water solution;

in this step, the pH is adjusted to 6 because gelatin is an amphoteric polymer having an isoelectric point of 5.0, and gelatin molecules exhibit negative charges, i.e., -NH, at a pH above its isoelectric point ₃ ⁺ Having a portion with-OH ^- Combined to form-NH ₂ thus-COO in the gelatin molecule ^- (negative charge) content greater than-NH ₃ ⁺ (positive charge) content, the molecule shows negative charge. When gelatin is in a medium below the isoelectric point, the gelatin molecule appears positively, i.e., -COO ^- Having a portion of and-H ⁺ The binding is converted into-COOH, thereby forming-NH in the gelatin molecule ₃ ⁺ (Positive charge) content greater than-COO ^- (negative charge) content, the molecule is positively charged. Therefore, the pH value of the system is adjusted to 6, the gelatin is negatively charged, the chitosan is positively charged due to protonation of free ammonia genes on the molecules in an acidic medium, and the negatively charged gelatin and the positively charged chitosan are subjected to complex coacervation reaction due to electrostatic interaction. The ionization reaction of gelatin at different pH values is shown as follows:

the protonation reaction process of chitosan in an acidic medium is shown as a formula III:

the complex coacervation reaction of gelatin and chitosan is shown as formula IV:

c. adding 270mL of corn oil into a three-neck flask, adding 5.00g of soybean phospholipid, heating to 36 ℃, and uniformly stirring, wherein the soybean phospholipid is an amphoteric surfactant which can be extracted from soybeans and is natural and nontoxic;

d. adding gelatin/chitosan mixed solution into corn oil for emulsification at 600rpm at 37 deg.C for 60 min;

e. turning off heating, and naturally cooling to room temperature; gelatin can undergo sol-gel transition, and when the temperature is higher than 35 ℃, gelatin swells, and when the temperature is lower than 35 ℃, gelatin gels. Thereby reducing the temperature to room temperature, being beneficial to the particles to form a relatively fixed shell membrane due to gelatin gel, improving the stability of the particles and being beneficial to the next step of crosslinking reaction;

f. and e, adding 3.50mL of 0.5% genipin solution into the cooled reaction system in the step e, reacting for 3h at room temperature, and then heating to 35 ℃ for reacting for 15 h.

In the step, genipin can generate a crosslinking reaction with a free amino-containing polymer, free amino groups on chitosan and gelatin attack olefinic carbon atoms at the C-3 position of genipin in an affinity manner under an acidic condition, and a dihydropyran ring is opened to form heterocyclic amine; in addition, ester group on genipin can generate SN with amino ₂ Nucleophilic substitution reaction to form amide and release methanol, so as to form a three-dimensional network structure polymer taking short-chain genipin as a cross-linking bridge; the cross-linking reaction process of genipin and chitosan is shown as formula V:

g. adding 1% of 1mL citric acid aqueous solution into the reaction system obtained after the step f, adding glacial acetic acid, adjusting the pH to 2-3, heating the reaction temperature to 40 ℃ under the protection of N2, reacting for 8h, and cooling to room temperature to obtain microcapsules with core substances of polyhexamethylene guanidine phosphate and benzalkonium chloride aqueous solution;

in the step, genipin reacts with free amino groups of gelatin and chitosan to generate crosslinking, free hydroxyl groups also exist on molecules of the chitosan and the gelatin, citric acid is added into a reaction system, under a certain condition, carboxyl groups on the citric acid and the free hydroxyl groups on macromolecules undergo esterification reaction, and the microcapsules taking the gelatin and the chitosan as capsule walls are subjected to secondary crosslinking, wherein the structural formula is shown as formula I:

h. standing the microcapsule obtained by the reaction for 2h, depositing the gelatin/chitosan microcapsule on the lower layer, depositing the corn oil on the upper layer, pouring out the upper oil phase, taking out the lower microcapsule, centrifuging, and separating out the oil phase to obtain the microcapsule with the core materials of polyhexamethylene guanidine phosphate and benzalkonium chloride aqueous solution. Finally transferring the microcapsule into a wide-mouth bottle, and sealing and storing;

(3) and (2) weighing microcapsules of which the capsule core substances are polyhexamethylene guanidine phosphate and benzalkonium chloride water solution, adding the microcapsules into the system obtained in the step (1), and stirring for 20-30min to obtain the disinfectant.

Example 3 thermogravimetric analysis (TG)

3.1 analytical methods

Weighing 2-6mg of sample, testing by adopting a TG/DSC synchronous thermal analyzer, heating from room temperature to 600 ℃ at the speed of 10 ℃/min, and taking nitrogen as gas atmosphere.

3.2 results of analysis

The thermal decomposition temperatures of the different samples are shown in table 2,

TABLE 2 thermal decomposition temperatures of different samples

Fig. 2 is a thermogravimetric graph of gelatin, chitosan, a gelatin/chitosan compound, genipin cross-linked gelatin/chitosan and citric acid secondary cross-linked microcapsule, and it can be seen from table 2 and fig. 2 that the thermal decomposition temperature of the microcapsule obtained by citric acid secondary cross-linking is the maximum and reaches 293 ℃, and the thermal stability is higher than that of genipin primary cross-linked gelatin/chitosan. Therefore, the microcapsule wall subjected to secondary citric acid crosslinking has higher strength and better thermal stability.

The reason for the above results is: the microcapsule subjected to secondary citric acid crosslinking is a double crosslinking agent, genipin is adopted for primary crosslinking, and genipin is a product obtained by hydrolyzing geniposide by beta-glucosidase and is an excellent natural biological crosslinking agent. The second cross-linking adopts citric acid which naturally exists in fruits such as citrus limon and the like, one citric acid molecule contains three carboxyl groups and one hydroxyl group, under certain reaction conditions, the citric acid can be subjected to esterification reaction with the hydroxyl groups on the gelatin and the chitosan, and the gelatin/chitosan on the capsule wall of the microcapsule can be further subjected to cross-linking and curing, so that the strength of the capsule wall is improved, and the thermal stability of the microcapsule is increased.

Example 4 optical microscopy characterization

The glass slide was then observed under an optical microscope of the type WV-CP240/G by sucking an appropriate amount of the disinfectant prepared in example 2 with a pipette and photographed and recorded.

FIG. 3 shows the microscopic appearance of the microcapsules of example 2, in which the core material is an aqueous solution of polyhexamethylene guanidine phosphate and benzalkonium chloride, and as can be seen from FIG. 3, the particle size of the microparticles gradually decreases and the particle size distribution becomes narrow with the increase of the emulsification time in the emulsification stage, and when the emulsification time reaches 60min, the particle size is smaller and the stability is better, and when the emulsification time continues to be prolonged, the particle size distribution of some microparticles becomes larger, so the emulsification time is preferably 60 min. After emulsification is finished, adding a cross-linking agent, and fully reacting to obtain the stable microcapsule with the core substance of polyhexamethylene guanidine phosphate and benzalkonium chloride aqueous solution.

A small amount of the microcapsules was placed on a glass slide and cut with another glass slide, and the microscopic view of the microcapsules after cutting is shown in FIG. 4. it can be seen from FIG. 4 that the microcapsules were ruptured and the cores flowed out after cutting. Therefore, when the microcapsules are subjected to shearing force, the microcapsules can be broken, and the polyhexamethylene guanidine phosphate and benzalkonium chloride aqueous solution flow out to complete the site-specific disinfection.

Example 5 bacterial experiments

The experimental method comprises the following steps:

1. primary reagent

Tryptone; extracting yeast powder; agar powder; sodium chloride; escherichia coli; staphylococcus aureus bacteria;

2. main instrument

A vertical pressure steam sterilizer; baking oven; an ultra-clean bench; a turbidimetric tube; a liquid transferring gun; a culture dish; an alcohol lamp; a conical flask; a centrifuge; centrifuge tubes and the like

3. The main steps of

LB medium:

liquid culture medium:

weighing 10g of tryptone, 5g of yeast extract powder, 10g of sodium chloride and 1000mL of water, adding the weighed materials into a conical flask, stirring by a glass rod to fully dissolve, sealing the opening of the conical flask by a sealing film after the materials are dissolved, putting the conical flask into a vertical pressure steam sterilizer, and sterilizing for 25-30min at 115 ℃.

Solid medium:

weighing 10g of tryptone, 5g of yeast extract powder, 20g of agar powder, 10g of sodium chloride and 1000mL of water, adding into a conical flask, stirring by a glass rod under heating to fully dissolve, sealing the conical flask with a sealing film after dissolving, placing into a vertical pressure steam sterilizer, and sterilizing at 115 ℃ for 25-30min

② inoculating bacteria:

firing the inoculating loop, slightly touching the original seed by the inoculating loop, inoculating into a culture medium, culturing at the constant temperature of 37 ℃ for 18h, and marking.

Thirdly, 10mL of uncooled solid culture medium is taken by a pipette and put into a culture dish, and the uncooled solid culture medium is uniformly spread and naturally cooled to form the solid culture medium.

Subpackaging, centrifuging and spreading bacteria

Place the tubes and add 1mL of the bacteria and media mixture (liquid from previous step) to each tube and centrifuge the numbers. Centrifuging, removing supernatant, adding 1mL of normal saline, comparing with a turbidimetric tube, and diluting with normal saline by corresponding times to obtain bacteria with required concentration.

Fifthly, taking 200 mu L of bacterial liquid, beating the bacterial liquid to one corner of a solid culture medium, then spreading the bacteria by a bacteria spreader with the surface facing upwards and the bottom marked, and putting the bacteria into an incubator at 37 ℃ for culturing for 18 hours.

Sixthly, observing whether a bacteriostatic circle exists on the solid culture medium after 18 hours and taking a picture.

The experimental results are as follows:

the bactericidal effect of the disinfectant prepared in example 2 on staphylococcus aureus is shown in fig. 5, and the bactericidal effect of the disinfectant prepared in example 2 on escherichia coli is shown in fig. 6. In fig. 5, the uppermost zone is the zone of inhibition by the non-microencapsulated germicidal sterilant, the middle zone is the zone of inhibition by the microencapsulated and ground germicidal sterilant, and the lowermost zone is the zone of inhibition by the microencapsulated but non-ground germicidal sterilant. As can be seen from fig. 5, the maximal bactericidal performance of the zone of inhibition generated by the non-microencapsulated bactericidal disinfectant is the best, the zone of inhibition of the microencapsulated and ground bactericidal disinfectant is smaller than that of the non-microencapsulated disinfectant, and the bactericidal performance is reduced because the grinding can not ensure that the microcapsules are completely broken, while the zone of inhibition generated by the microencapsulated and non-ground bactericidal disinfectant is almost not generated due to the coating of the wall material. FIG. 6 shows the bactericidal effect on Escherichia coli, the uppermost zone is the zone of inhibition by the microencapsulated but unground bactericidal disinfectant, the middle zone is the zone of inhibition by the ungelled bactericidal disinfectant, and the lowermost zone is the zone of inhibition by the microencapsulated and ground bactericidal disinfectant. As can be seen from FIG. 6, the bactericidal activity was comparable to that of Staphylococcus aureus.

The experiment can prove that: (1) the bactericidal disinfectant has good bactericidal effect on staphylococcus aureus and escherichia coli; (2) the sterilization disinfectant can be successfully coated by microencapsulation; (3) the microcapsule is broken after being stressed, and the capsule core substance has bactericidal effect after flowing out.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.

Claims

1. A germicidal sterilant, comprising: the microcapsule comprises a microcapsule with a core material of polyhexamethylene guanidine hydrochloride (phosphate) and a benzalkonium chloride (bromide) aqueous solution, wherein the wall material of the microcapsule is a polymer formed by crosslinking citric acid/genipin and gelatin/chitosan, and the chemical structural formula of the polymer is shown as the formula (I):

in the formula (I), R1-R9 are selected from one of residues of eighteen different amino acids of glycine, alanine, serine, aspartic acid, glutamic acid amino, proline, arginine, histidine, tyrosine, cystine, leucine, threonine, methionine, valine, phenylalanine, tryptophan, glutamic acid and lysine.

2. The germicidal sterilant according to claim 1, wherein: in the formula (I):

r3, R7 are residues of lysine or arginine;

r2, R4 and R8 are residues of aspartic acid or glutamic acid amino;

r1, R9 are residues of serine, threonine or tyrosine.

3. A preparation method of a sterilizing disinfectant specifically comprises the following steps:

4. The preparation method of the sterilizing disinfectant as claimed in claim 3, wherein the ratio of the components in the step (1) by weight is as follows: 5-10 parts of glycerol, 5-10 parts of propylene glycol and 0.2-0.5 part of water-soluble cherry essence; the ratio of the total mass of the components in the step (1) to the total mass of the components in the step (2) is 1: 1.

5. A method of preparing a germicidal sterilant according to claim 3, further comprising, after step f: and f, standing the reaction system after the step f, pouring out an upper oil phase, centrifuging, and separating out the oil phase to obtain the microcapsule with the core substances of polyhexamethylene guanidine hydrochloride (phosphate) and benzalkonium chloride (bromide) aqueous solution.

6. A method of preparing a germicidal sterilant according to claim 3 wherein in step b the pH is preferably adjusted to 6.

7. A method for preparing a germicidal disinfectant as claimed in claim 3, wherein the step e comprises the following steps: and d, adding 0.5% genipin aqueous solution into the solution system obtained in the step d, reacting for 3 hours at room temperature, heating to 35 ℃, reacting for 15 hours again, and fully reacting.

8. The method for preparing the sterilizing disinfectant as claimed in claim 3, wherein the specific steps of the step f are as follows: and e, adding a 1% citric acid aqueous solution into the reaction system in the step e, adding glacial acetic acid, adjusting the pH value to 2-3, raising the reaction temperature to 40 ℃ under the protection of N2, reacting for 8 hours, and cooling to room temperature to obtain the microcapsule with the capsule core substances of polyhexamethylene guanidine hydrochloride (phosphate) and benzalkonium chloride (bromide) aqueous solution.

9. The method according to claim 3, wherein the vegetable oil of step c is one or more of corn oil, olive oil, soybean oil and peanut oil.

10. The bactericidal disinfectant microcapsule as claimed in claim 3, wherein the mixture ratio of the components is as follows by weight: 45-55 parts of gelatin, 3-7 parts of chitosan, 350 parts of 1.0% acetic acid solution 250-ion, 40-60 parts of soybean lecithin, 30-40 parts of 0.5% genipin aqueous solution, 5-15 parts of 1.0% citric acid solution and 3000 parts of vegetable oil 2500-ion.