CN110240664B - Preparation method of antibacterial chitosan quaternary ammonium salt and product - Google Patents
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Abstract
The invention provides a preparation method of an antibacterial chitosan quaternary ammonium salt, which comprises the following steps: 1) preparing an aqueous solution of chitosan; 2) adding long alkyl chain epoxy quaternary ammonium salt and short alkyl chain epoxy quaternary ammonium salt for reaction; 3) precipitating the reaction solution obtained in the step 2), washing and drying to obtain a product. By grafting the long alkyl chain epoxy quaternary ammonium salt and the alkyl chain epoxy quaternary ammonium salt on the chitosan together, the antibacterial chitosan has stronger electropositivity and can be better combined with a negatively charged bacterial membrane, thereby improving the antibacterial effect. The co-grafting proportion of the chitosan and the chitosan is optimized and adjusted, so that the biocompatibility of the antibacterial chitosan can be effectively improved while the antibacterial effect is ensured.
Description
Technical Field
The invention belongs to the field of antibacterial materials, and relates to a preparation method and a product of antibacterial chitosan quaternary ammonium salt.
Background
Chitosan is a deacetylated product of chitin, and is widely applied to the fields of medicine, food, daily chemicals, textile, agriculture and the like due to the regenerability, good biocompatibility and a plurality of unique physiological and pharmacological functions, so that the chitosan has wide application and development prospects. The Chinese patent publication No. CN101929074A discloses a method for preparing a chitosan quaternary ammonium salt, which is characterized in that C4-18 alkyl dimethyl tertiary amine is firstly used for reacting with epoxy chloropropane to obtain a long alkyl chain epoxy quaternary ammonium salt, then the long alkyl chain epoxy quaternary ammonium salt is used for carrying out an epoxy ring-opening reaction with chitosan, and the quaternary ammonium salt is grafted on the chitosan, so that the material has a certain antibacterial capacity. However, the single long alkyl chain epoxy quaternary ammonium salt has poor biocompatibility, can cause serious cytotoxicity and hemolysis, cannot be used in the field of contact with human tissues, and has certain limitation in application. Therefore, it is required to provide an antibacterial chitosan material having excellent antibacterial effect and good biocompatibility.
Disclosure of Invention
In view of the above, the present invention provides a method for preparing an antibacterial chitosan quaternary ammonium salt and a product thereof.
The invention specifically provides the following technical scheme:
1. a preparation method of antibacterial chitosan quaternary ammonium salt is characterized by comprising the following steps:
1) preparing an aqueous solution of chitosan;
2) adding long alkyl chain epoxy quaternary ammonium salt and short alkyl chain epoxy quaternary ammonium salt for reaction;
3) precipitating the reaction solution obtained in the step 2), washing and drying to obtain a product.
Further, before the step 3), adjusting the pH value of the reaction solution to 10-13 to separate out a product, filtering, and performing subsequent precipitation washing on the filtered product.
Further, the mass ratio of the long alkyl chain epoxy quaternary ammonium salt to the chitosan is 0.1-3:1, and the mass ratio of the long alkyl chain epoxy quaternary ammonium salt to the short alkyl chain epoxy quaternary ammonium salt is 0.5: 1-3: 1.
Further, the aqueous solution of chitosan in step 1) is aqueous solution of chitosan hydrochloride, chitosan acetate, chitosan lactate, chitosan citrate and carboxymethyl chitosan or dilute solution of chitosan dissolved in hydrochloric acid, acetic acid, lactic acid and citric acid.
Further, the long alkyl chain epoxy quaternary ammonium salt in the step 2) has a structural formula:
n is 6-14, and the structural formula of the short alkyl chain epoxy quaternary ammonium salt is as follows:
n＝1-4。
further, the long alkyl chain epoxy quaternary ammonium salt in the step 2) is a precipitation product of solvent-free reaction of epoxy chloropropane and N, N-dimethyl X alkyl tertiary amine, wherein X is 6-16; the short alkyl chain epoxy quaternary ammonium salt is a precipitation product of solvent-free reaction of epichlorohydrin and N, N-dimethyl Y alkyl tertiary amine, wherein Y is 2-4 or 2, 3-epoxypropyl trimethyl ammonium chloride; the reaction temperature is 40-60 ℃, the reaction time is 1-24 hours, and the precipitating agent for precipitation is petroleum ether, diethyl ether, cyclohexane or normal hexane.
Further, the pH value of the chitosan aqueous solution in the step 1) is 3-7, the reaction temperature in the step 2) is 50-70 ℃, and the reaction time is 12-120 hours.
Further, the precipitating agent for precipitation in the step 3) is one or more of ethanol, methanol and acetone, and the volume ratio of the precipitating agent to the reaction liquid is 4-20: 1.
2. the antibacterial chitosan quaternary ammonium salt obtained by the preparation method has the molecular formula:
3. the antibacterial chitosan quaternary ammonium salt obtained by the preparation method has the molecular formula:
the invention has the beneficial effects that:
(1) short alkyl chain epoxy quaternary ammonium salt is independently grafted on chitosan, and the antibacterial performance is poor due to the lack of strong hydrophobicity and the fact that long alkyl chains of bacterial cell membranes can be damaged. According to the invention, the long alkyl chain epoxy quaternary ammonium salt and the short alkyl chain epoxy quaternary ammonium salt with excellent antibacterial performance are grafted on the chitosan simultaneously, so that the antibacterial chitosan has stronger electropositivity and can be better combined with a negatively charged bacterial membrane, thereby improving the antibacterial effect.
(2) Although the long alkyl chain epoxy quaternary ammonium salt has better antibacterial effect, the long alkyl chain epoxy quaternary ammonium salt also has larger destructive effect on common cells, and the single grafting of the long alkyl chain epoxy quaternary ammonium salt on the chitosan shows that the biocompatibility is low, namely hemolysis; and the short alkyl chain epoxy quaternary ammonium salt has good biocompatibility. Therefore, the co-grafting proportion of the two is optimized and adjusted, so that the biocompatibility and the antibacterial property of the antibacterial chitosan can be effectively optimized; and the ratio of the two must be within a suitable range to achieve the best results. The biological compatibility of the chitosan quaternary ammonium salt is influenced by too low feeding/grafting amount of the short alkyl chain epoxy quaternary ammonium salt, and the antibacterial efficiency of the chitosan quaternary ammonium salt is influenced by too high grafting amount.
Detailed Description
The following describes in detail preferred embodiments of the present invention.
Example 1
The synthesis of the antibacterial chitosan quaternary ammonium salt X1 comprises the following steps: :
1) dropwise adding 3g of epichlorohydrin into 4g N, N-dimethylhexylamine, stirring for 2h at 55 ℃, after the reaction is finished, precipitating and washing with petroleum ether for 2 times, and drying under reduced pressure to obtain the epoxy quaternary ammonium salt A1 (2-oxaanemethanaminium, N-hexane-N, N-dimethyl-), of which the molecular formula is long-chain alkyl chain
2) Dripping 1.0g of epichlorohydrin into 1.0g of 1.0g N, N-dimethylethylamine, stirring at 50 ℃ for 36 hours, washing with diethyl ether precipitate for 2 times, and drying under reduced pressure to obtain short-chain alkyl epoxy quaternary ammonium salt B1 with the molecular formula of
3) 10g of chitosan hydrochloride (deacetylation degree 85%, molecular weight 8 ten thousand) is added into 2L of water, the mixture is heated and dissolved at 60 ℃, hydrochloric acid is added to adjust the pH to 4, then 15g of epoxy quaternary ammonium salt A1 and 7.5g of epoxy quaternary ammonium salt B1 are dissolved in 50mL of deionized water and added into the chitosan solution, and the temperature is controlled at 60 ℃ to react for 36 hours.
4) After the reaction is finished, ethanol is used for precipitation and washing for three times, and reduced pressure drying is carried out, so as to obtain the antibacterial chitosan quaternary ammonium salt X1.
Example 2
The synthesis of the antibacterial chitosan quaternary ammonium salt X2 comprises the following steps: :
1) dropwise adding 2g of epoxy chloropropane into 4g N, N-dimethyl octylamine, stirring for 24h at 40 ℃, after the reaction is finished, carrying out ether precipitation and washing for 2 times, and drying under reduced pressure to obtain the long alkyl chain epoxy quaternary ammonium salt A2(2-Oxiranemethanaminium, N-octyl-N, N-dimethyl-), wherein the molecular formula is shown in the specification
2) Dripping 2.0g of epichlorohydrin into 5.0g N, N-dimethylpropylamine, stirring at 40 deg.C for 6 hr, washing with diethyl ether precipitate for 2 times, and drying under reduced pressure to obtain epoxy quaternary ammonium salt B2 with molecular formula of
3) 5g of chitosan citrate (degree of deacetylation 85%, molecular weight 8 ten thousand) was added to 500mL of water, and dissolved by heating at 70 ℃ with citric acid to pH 5. Then 7.5g of epoxy quaternary ammonium salt A2 and 3g of epoxy quaternary ammonium salt B2 were dissolved in 20mL of deionized water and added to the chitosan solution, and the temperature was controlled at 70 ℃ for reaction for 24 h.
4) After the reaction is finished, ethanol is used for precipitation and washing for 1 time, acetone is used for washing for 2 times, and then vacuum drying is carried out, so that the antibacterial chitosan quaternary ammonium salt X2 can be obtained.
Example 3
The synthesis of the antibacterial chitosan quaternary ammonium salt X3 comprises the following steps:
1) dropwise adding 3g of epichlorohydrin into 5g N, N-dimethyl decylamine, heating and stirring for 3h at 50 ℃, precipitating and washing the reaction solution with N-hexane for three times after the reaction is finished, and then carrying out vacuum drying to obtain a quaternizing agent A3 (2-oxaranine, N-decyl-N, N-dimethyl-), wherein the molecular formula is as follows:
2) dripping 1.0g of epichlorohydrin into 4.0g of 4.0g N, N-dimethylbutylamine, stirring for 4 hours at 40 ℃, washing for 2 times by ether precipitation after the end, and drying under reduced pressure to obtain epoxy quaternary ammonium salt B3 with the molecular formula of
3) 5g of chitosan hydrochloride (degree of deacetylation 90%, molecular weight 3 ten thousand) was added to 500mL of deionized water, and dissolved by heating at 50 ℃ to adjust pH to 3. Then 4g of epoxy quaternary ammonium salt A3 and 2g of epoxy quaternary ammonium salt B3 are dissolved in 20mL of deionized water and added into the chitosan solution, the temperature is controlled at 50 ℃, and the mixture is stirred and reacts for 72 hours;
3) and after the reaction is finished, adding solid NaOH into the reaction solution to adjust the pH value of the system to be 12, separating out chitosan, carrying out suction filtration on the separated out product, then precipitating and washing the precipitate with ethanol for three times, and drying the precipitate under reduced pressure to obtain the antibacterial chitosan quaternary ammonium salt X3.
Example 4
The preparation method X4 of the synthetic antibacterial chitosan quaternary ammonium salt comprises the following steps:
1) dropwise adding 3g of epoxy chloropropane into 5g N, N-dimethyldodecylamine, heating and stirring for 3h at 50 ℃, precipitating and washing the reaction solution for three times by using N-hexane after the reaction is finished, and then carrying out vacuum drying to obtain a quaternizing agent A4 (2-oxaranine, N-dodecyl-N, N-dimethyl-), wherein the molecular formula is as follows:
2) preparation of epoxy Quaternary ammonium salt B2, molecular formula
3) 15g of chitosan (degree of deacetylation 85%, molecular weight 8 ten thousand) was added to 2L of an aqueous hydrochloric acid solution (mass fraction 0.1%), and dissolved by heating at 55 ℃ to adjust pH 5. Then 8g of epoxy quaternary ammonium salt A4 and 6g of epoxy quaternary ammonium salt B2 are dissolved in 40mL of deionized water and added into the chitosan solution, the temperature is controlled at 55 ℃, and the mixture is stirred and reacts for 60 hours;
3) after the reaction is finished, precipitating and washing the product by using methanol for 1 time, then washing the product by using acetone for 2 times, and then drying the product in vacuum to obtain the antibacterial chitosan quaternary ammonium salt X4.
Example 5
The preparation method of the synthetic antibacterial chitosan quaternary ammonium salt comprises the following steps
1) Dropwise adding 2g of epichlorohydrin into 5g N, N-dimethyltetradecylamine, heating and stirring for 2h at 50 ℃, precipitating and washing the reaction liquid for three times by using cyclohexane after the reaction is finished, and then drying in vacuum to obtain a quaternizing agent A5 (2-oxaranine, N-tetradecyl-N, N-dimethyltetramine-), wherein the molecular formula is shown in the specification
2) 10g of chitosan lactate (degree of deacetylation 90%, molecular weight 5 ten thousand) was added to 750mL of deionized water, dissolved by heating at 65 ℃, and then the system pH was adjusted to 4 with hydrochloric acid. Then 6g of epoxy quaternary ammonium salt A5 and 2g of 2, 3-epoxypropyl trimethyl ammonium chloride are dissolved in 30mL of deionized water and added into the chitosan solution, the temperature is controlled at 55 ℃, and the mixture is stirred and reacted for 48 hours;
3) and (3) after the reaction is finished, adding solid KOH into the reaction solution to adjust the pH value of the system to be 10, separating out chitosan, carrying out suction filtration on the separated out product, then carrying out precipitation washing for three times by using ethanol, and drying under reduced pressure to obtain the antibacterial chitosan quaternary ammonium salt X5.
Example 6
The synthesis of the antibacterial chitosan quaternary ammonium salt X6 comprises the following steps:
1) preparation of epoxy Quaternary ammonium salt A1, molecular formula
2) Preparation of epoxy Quaternary ammonium salt B2, molecular formula
3) 10g of carboxymethyl chitosan (degree of deacetylation 85%, degree of substitution with carboxymethyl 75%, molecular weight 5 ten thousand) was added to 1L of deionized water, and dissolved by heating at 60 ℃ with hydrochloric acid to adjust pH 6. Then 3g of epoxy quaternary ammonium salt A1 and 2g of epoxy quaternary ammonium salt B2 are dissolved in 15mL of deionized water and added into the carboxymethyl chitosan solution, and the temperature is controlled at 60 ℃ for reaction for 48 hours;
4) after the reaction is finished, ethanol is used for precipitation and washing for three times, and reduced pressure drying is carried out, so that the antibacterial chitosan quaternary ammonium salt X6 can be obtained.
Comparative example 1
1) Preparation of epoxy Quaternary ammonium salt A1, molecular formula
2) 10g of chitosan (deacetylation degree of 85%, molecular weight of 8 ten thousand) is added into 2L of water, the mixture is heated and dissolved at 60 ℃, hydrochloric acid is added to adjust the pH to 4, then 15g of epoxy quaternary ammonium salt A1 is dissolved in 50mL of deionized water and added into the chitosan solution, and the temperature is controlled to be 60 ℃ for reaction for 36 hours.
3) After the reaction is finished, ethanol is used for precipitation and washing for three times, and reduced pressure drying is carried out, so as to obtain the antibacterial chitosan quaternary ammonium salt Y1.
Comparative example 2
1) Preparation of epoxy Quaternary ammonium salt B2, molecular formula
2) 15g of chitosan (degree of deacetylation 85%, molecular weight 8 ten thousand) was added to 2L of an aqueous hydrochloric acid solution (mass fraction 0.1%), and dissolved by heating at 55 ℃ to adjust pH 5. Then 6g of epoxy quaternary ammonium salt B2 is dissolved in 40mL of deionized water and added into the chitosan solution, the temperature is controlled at 55 ℃, and the stirring reaction is carried out for 60 hours;
3) after the reaction is finished, precipitating and washing the product by using methanol for 1 time, then washing the product by using acetone for 2 times, and then drying the product in vacuum to obtain the antibacterial chitosan quaternary ammonium salt Y2.
Comparative example 3
1) Epoxy quaternary ammonium salt A1 with long-chain alkyl chain and molecular formula of
2) The epoxy quaternary ammonium salt B1 with short chain alkyl is obtained by the step 2 in the example 1, and the molecular formula is
3) 10g of chitosan hydrochloride (deacetylation degree of 85%, molecular weight of 8 ten thousand) is added into 2L of water, the mixture is heated and dissolved at 60 ℃, hydrochloric acid is added to adjust the pH to 4, then 6g of epoxy quaternary ammonium salt A1 and 15g of epoxy quaternary ammonium salt B1 are dissolved in 50mL of deionized water and added into the chitosan solution, and the temperature is controlled to be 60 ℃ for reaction for 36 hours.
4) After the reaction is finished, ethanol is used for precipitation and washing for three times, and reduced pressure drying is carried out, so as to obtain the antibacterial chitosan quaternary ammonium salt Y3.
Comparative example 4
1) The epoxy quaternium A2 was prepared as in example 2, step 1, having the formula
2) 5g of chitosan citrate (degree of deacetylation 85%, molecular weight 8 ten thousand) was added to 500mL of water, and dissolved by heating at 70 ℃ with citric acid to pH 5. Then 7.5g of epoxy quaternary ammonium salt A2 and 3g of epoxy quaternary ammonium salt B2 are dissolved in 20mL of deionized water and added into the chitosan solution, and the temperature is controlled to be 70 ℃ for reaction for 24h
3) After the reaction is finished, ethanol is used for precipitation and washing for 1 time, acetone is used for washing for 2 times, and then vacuum drying is carried out, so that the antibacterial chitosan quaternary ammonium salt Y4 is obtained.
Comparative example 5
1) Preparation of epoxy Quaternary ammonium salt A4, molecular formula
2) 15g of chitosan (degree of deacetylation 85%, molecular weight 8 ten thousand) was added to 2L of an aqueous hydrochloric acid solution (mass fraction 0.1%), and dissolved by heating at 55 ℃ to adjust pH 5. Then 8g of epoxy quaternary ammonium salt A4 is dissolved in 40mL of deionized water and added into the chitosan solution, the temperature is controlled at 55 ℃, and the stirring reaction is carried out for 60 hours;
3) after the reaction is finished, precipitating and washing the product by using methanol for 1 time, then washing the product by using acetone for 2 times, and then drying the product in vacuum to obtain the antibacterial chitosan quaternary ammonium salt Y5.
Comparative example 6
1) Preparation of epoxy Quaternary ammonium salt A5, molecular formula
2) 10g of chitosan lactate (degree of deacetylation 90%, molecular weight 5 ten thousand) was added to 750mL of deionized water, dissolved by heating at 65 ℃, and then the system pH was adjusted to 4 with hydrochloric acid. Then 6g of epoxy quaternary ammonium salt A5 is dissolved in 30mL of deionized water and added into the chitosan solution, the temperature is controlled at 55 ℃, and the stirring reaction is carried out for 48 hours;
3) and after the reaction is finished, adding solid KOH into the reaction solution to adjust the pH value of the system to be 10, separating out chitosan, carrying out suction filtration on the separated out product, then carrying out precipitation washing for three times by using ethanol, and drying under reduced pressure to obtain the antibacterial chitosan quaternary ammonium salt Y6.
Comparative example 7
1) Epoxy quaternary ammonium salt A1 with long-chain alkyl chain and molecular formula of
2) The epoxy quaternary ammonium salt B1 with short chain alkyl is obtained by the step 2 in the example 1, and the molecular formula is
3) 10g of chitosan hydrochloride (deacetylation degree 85%, molecular weight 8 ten thousand) is added into 2L of water, heated and dissolved at 60 ℃, hydrochloric acid is added to adjust the pH to 4, and then 7.5g of epoxy quaternary ammonium salt A1 and 1.5g of epoxy quaternary ammonium salt B1 are dissolved in 50mL of deionized water and added into the chitosan solution, and the temperature is controlled at 60 ℃ to react for 36 h.
4) After the reaction is finished, ethanol is used for precipitation and washing for three times, and reduced pressure drying is carried out, so as to obtain the antibacterial chitosan quaternary ammonium salt Y7.
Test example 1 comparison of antibacterial Effect
Detection conditions are as follows: the antibacterial performance test of staphylococcus aureus was performed on experimental examples 1-6, comparative examples 1-9, chitosan (degree of deacetylation 85%, molecular weight 8 ten thousand) and carboxymethyl chitosan (degree of deacetylation 85%, degree of carboxymethyl substitution 75%, molecular weight 5 ten thousand). The bacteria for detection is staphylococcus aureus ATCC 25923.
The detection method refers to national standard GB15979-2002, and the tested material is prepared into 1.5mg/mL aqueous solution or dispersion (sterilized), each tube is 5mL, and the control group is sterile water. Diluting cultured Staphylococcus aureus with PBS to 1 × 10 concentration6CFU/mL of bacterial suspension. And (3) taking the suspension liquid, dropwise adding 100 mu L of the suspension liquid into each tube of sample liquid and the control sample liquid, and uniformly mixing. At this point, the time was started and the effect was 5 min. Sucking 0.5mL of sample solution from each tube and adding the sample solution into 5mL of PBS respectively, fully mixing the sample solution and the control sample solution uniformly, diluting the mixture properly, putting 0.5mL of the mixture into a plate, adding 15mL of nutrient agar culture medium cooled to 40-45 ℃, and rotating the plate to fully mix the mixture uniformly. Cultured at 37 ℃ for 48h, and viable colonies were counted. The experiment was repeated three times and the inhibition was calculated as follows:
x ═ a-B)/a × 100%, where: x-bacteriostasis rate,% A-average colony number of control sample B-average colony number of tested sample, and obtaining the results of the average bacteriostasis rate of each material as follows:
table 1 average bacteriostasis test
As can be seen from Table 1, the bacteriostasis rates of X1-X6 are much higher than those of Y1-Y6. X1-X6 are antibacterial chitosan with co-grafted long alkyl chain quaternary ammonium salt and short alkyl chain quaternary ammonium salt; y1, Y4-Y6 are epoxy quaternary ammonium salts only with lengthened alkyl chains and without short alkyl chains; y2 is epoxy quaternary ammonium salt grafted with only short alkyl chain and not lengthened with alkyl chain; y3 and Y7 are proportioned charges that are not provided in accordance with the claimed invention.
Therefore, the antibacterial effect of the antibacterial chitosan synthesized by the antibacterial chitosan co-grafted with the long alkyl chain quaternary ammonium salt and the short alkyl chain quaternary ammonium salt is obviously better than that of the antibacterial chitosan synthesized by single grafting, and the grafting product with the preferable ratio of the long alkyl chain quaternary ammonium salt to the short alkyl chain quaternary ammonium salt is most preferable.
Test example 2 hemolytic effect comparison
The test method comprises the following steps: washing and centrifuging collected fresh rabbit blood for three times by using normal saline, removing supernatant to obtain erythrocyte suspension, and preparing the erythrocyte suspension with the volume fraction of 4% by using the normal saline. The physiological saline solution or dispersion liquid of the experimental examples X1-X6 and the comparative examples Y1-Y9 with the concentration of 4mg/mL, 2mg/mL, 1mg/mL and 0.5mg/mL is prepared respectively, the volume is 0.5mL, 5 parallel samples are arranged in each group of samples, the negative control group is physiological saline, and the positive control group is Triton-X reagent with the mass fraction of 1%. The 4% erythrocyte suspension and the material solution were mixed in equal volumes and then left at 37 ℃ for 3 hours. After 3h, the sample is taken out and centrifuged at 3000rpm, 140 mu L of supernatant of each sample is added into a 96-well plate, the absorption light intensity of each well at 545nm is read by a microplate reader, and the hemolysis rate is calculated. The hemolysis results are as follows:
TABLE 2 hemolytic Effect test
As shown in the table above, the hemolysis rates of X1-X6 are very small, the hemolysis rates of Y1, Y4-Y6 are very large, Y1, Y4-Y6 are epoxy quaternary ammonium salts with only lengthened alkyl chains, and short alkyl chains are not added. The hemolysis rates of Y3 and Y7 were also higher, with Y3 and Y7 being dosed at a ratio not provided by the claimed protocol.
It can be seen from this that: only long alkyl chain epoxy quaternary ammonium salt is added, short alkyl chain epoxy quaternary ammonium salt is not added, so that the hemolysis rate of the material is too high, and the material has no medical practical value, therefore, the antibacterial chitosan co-grafted with the long alkyl chain quaternary ammonium salt and the short alkyl chain quaternary ammonium salt has obviously excellent blood compatibility, and the biocompatibility of the antibacterial chitosan can be effectively improved by adjusting the proportion of the long alkyl chain quaternary ammonium salt to the short alkyl chain quaternary ammonium salt.
Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the scope of the invention as defined by the appended claims.
Claims (6)
1. The application of the antibacterial chitosan quaternary ammonium salt as an antibacterial material is characterized by comprising the following preparation steps:
1) preparing an aqueous solution of chitosan;
2) adding long alkyl chain epoxy quaternary ammonium salt and short alkyl chain epoxy quaternary ammonium salt for reaction;
3) precipitating the reaction solution obtained in the step 2), washing and drying to obtain a product;
the mass ratio of the long alkyl chain epoxy quaternary ammonium salt to the chitosan is 0.1-3:1, and the mass ratio of the long alkyl chain epoxy quaternary ammonium salt to the short alkyl chain epoxy quaternary ammonium salt is 0.5: 1-3: 1;the structural formula of the long alkyl chain epoxy quaternary ammonium salt in the step 2) is as follows:
n is 6-14, and the structural formula of the short alkyl chain epoxy quaternary ammonium salt is as follows:
n＝1-4。
2. the use of the antibacterial chitosan quaternary ammonium salt as an antibacterial material according to claim 1, wherein: adjusting the pH value of the reaction solution to 10-13 before the step 3), separating out a product, filtering, and performing subsequent precipitation and washing on the filtered product.
3. The use of the antibacterial chitosan quaternary ammonium salt as an antibacterial material according to claim 1 or 2, wherein: the aqueous solution of chitosan in the step 1) is aqueous solution of chitosan hydrochloride, chitosan acetate, chitosan lactate, chitosan citrate and carboxymethyl chitosan or dilute solution of chitosan dissolved in hydrochloric acid, acetic acid, lactic acid and citric acid.
4. The use of the antibacterial chitosan quaternary ammonium salt as an antibacterial material according to claim 1 or 2, wherein: the long alkyl chain epoxy quaternary ammonium salt in the step 2) is a precipitation product of solvent-free reaction of epoxy chloropropane and N, N-dimethyl X alkyl tertiary amine, wherein X is 6-14; the short alkyl chain epoxy quaternary ammonium salt is a precipitation product of solvent-free reaction of epichlorohydrin and N, N-dimethyl Y alkyl tertiary amine, wherein Y is 2-4 or 2, 3-epoxypropyl trimethyl ammonium chloride; the reaction temperature is 40-60 ℃, the reaction time is 1-24 hours, and the precipitating agent for precipitation is petroleum ether, diethyl ether, cyclohexane or normal hexane.
5. The use of the antibacterial chitosan quaternary ammonium salt as an antibacterial material according to claim 1 or 2, wherein: the pH value of the chitosan aqueous solution obtained in the step 1) is 3-7, the reaction temperature in the step 2) is 50-70 ℃, and the reaction time is 12-120 hours.
6. The use of the antibacterial chitosan quaternary ammonium salt as an antibacterial material according to claim 1 or 2, wherein: the precipitator for precipitation in the step 3) is one or more of ethanol, methanol and acetone, and the volume ratio of the precipitator to the reaction liquid is (4-20): 1.
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| CN116410348A (en) * | 2023-04-03 | 2023-07-11 | 华侨大学 | Preparation method of chitosan quaternary ammonium salt |
| CN116655826A (en) * | 2023-07-26 | 2023-08-29 | 青岛溯博生物技术有限公司 | Cationic carboxymethyl chitosan |
| CN116891542B (en) * | 2023-09-11 | 2023-12-15 | 青岛溯博生物技术有限公司 | Cationic carboxymethyl chitosan with antibacterial effect and application thereof |
Family Cites Families (6)
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| DE3245784A1 (en) * | 1982-12-10 | 1984-06-14 | Wella Ag, 6100 Darmstadt | COSMETIC AGENT BASED ON QUATERNAUS CHITOSAN DERIVATIVES, NEW QUATERNAERE CHITOSAN DERIVATIVES AND METHOD FOR THE PRODUCTION THEREOF |
| DK0552176T3 (en) * | 1990-10-09 | 1996-03-04 | Pfizer | Quaternary ammonium salts of chitosan |
| CN101235099B (en) * | 2007-01-29 | 2011-01-12 | 北京理工大学 | Carboxymethyl chitosan quaternary ammonium salt derivatives and preparation method thereof |
| CN102382206B (en) * | 2010-08-30 | 2014-10-01 | 华东理工大学 | Chitooligosaccharide quaternary ammonium salt and preparation method thereof |
| CN106496358A (en) * | 2016-11-09 | 2017-03-15 | 深圳大学 | Amphiphilic chitosan quaternary ammonium salt derivatives and its preparation and application |
| CN107349461B (en) * | 2017-07-21 | 2019-10-29 | 北京化工大学 | A kind of preparation method and applications of antibacterial anti hemorrhagic chitosan |
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