阅读说明：本技术 一种环境友好型松香基季铵盐抗菌剂的制备方法 (Preparation method of environment-friendly rosinyl quaternary ammonium salt antibacterial agent ) 是由 唐艳军 黄梦乐 于 2021-08-05 设计创作，主要内容包括：本发明涉及一种环境友好型松香基季铵盐抗菌剂的制备方法,属于抗菌技术领域。本发明首先松香和环氧氯丙烷加入到三口烧瓶中,加热到60℃并使之溶解成均相后,再滴加催化剂四丁基溴化铵,得到3-松香酰氧-2-羟丙基氯黄色粘稠状液体；然后以3-松香酰氧-2-羟丙基氯和胺盐为原料,乙醇为溶剂,95℃下反应8小时,得到松香基胺盐；最后,将松香基胺盐和环氧氯丙烷加入到三口烧瓶中,在90℃下回流反应5h,得到松香基季铵盐抗菌剂。本发明一方面能充分的利用松香这一可再生资源,实现了对松香深加工产业的开发,另一方面为价格低廉、高活性、无毒、环保、易生物降解的抗菌剂的制备提供了新的路径和方法。(The invention relates to a preparation method of an environment-friendly rosin-based quaternary ammonium salt antibacterial agent, belonging to the technical field of antibiosis. Firstly, adding rosin and epoxy chloropropane into a three-neck flask, heating to 60 ℃, dissolving into a homogeneous phase, and then dropwise adding a catalyst tetrabutylammonium bromide to obtain 3-rosin acyloxy-2-hydroxypropyl chloride yellow viscous liquid; then 3-abietyloxy-2-hydroxypropyl chloride and amine salt are used as raw materials, ethanol is used as a solvent, and the reaction is carried out for 8 hours at the temperature of 95 ℃ to obtain abietylamine salt; and finally, adding the rosinyl amine salt and epoxy chloropropane into a three-neck flask, and carrying out reflux reaction for 5 hours at the temperature of 90 ℃ to obtain the rosinyl quaternary ammonium salt antibacterial agent. The invention can fully utilize the renewable resources of rosin to realize the development of rosin deep processing industry, and provides a new path and method for preparing the antibacterial agent which has low price, high activity, no toxicity, environmental protection and easy biodegradation.)

1. The preparation method of the environment-friendly rosinyl quaternary ammonium salt antibacterial agent is characterized by comprising the following steps: the method comprises the following steps:

(1) adding 5-7 parts by weight of rosin and 10-12 parts by weight of epoxy chloropropane into a three-neck flask, heating to 60 ℃, dissolving into a homogeneous phase, then dropwise adding 0.15-0.2 part by weight of tetrabutylammonium bromide serving as a catalyst, and heating to 95 ℃ for reaction for 5 hours;

after the reaction is finished, rotationally evaporating at 65 ℃ until no distillate exists to obtain 3-rosin acyl oxide-2-hydroxypropyl chloride yellow viscous liquid;

(2) 5-7 parts by weight of 3-abietyl oxide-2-hydroxypropyl chloride and 7.5-10.5 parts by weight of amine salt are taken as raw materials, ethanol is taken as a solvent, and the reaction is carried out for 8 hours at 95 ℃;

after the reaction is finished, removing ethanol and unreacted amine salt by rotary evaporation to obtain rosinyl amine salt;

(3) adding 5-7 parts by weight of rosinyl amine salt and 5-7 parts by weight of epoxy chloropropane into a three-neck flask, carrying out reflux reaction for 5 hours at 90 ℃, washing with petroleum ether and extracting with ether after the reaction is finished, thus obtaining the rosinyl quaternary ammonium salt antibacterial agent.

2. The method for preparing the environmentally-friendly rosinyl quaternary ammonium salt antibacterial agent according to claim 1, wherein the method comprises the following steps: in the step (2), the amine salt is one of triethylamine, diethylamine or di-n-butylamine.

3. The method for preparing the environmentally-friendly rosinyl quaternary ammonium salt antibacterial agent according to claim 1, wherein the method comprises the following steps: in the step (2), the molar ratio of the 3-abietyloxy-2-hydroxypropyl chloride to the amine salt is 1: 1.5.

4. the method for preparing the environmentally-friendly rosinyl quaternary ammonium salt antibacterial agent according to claim 1, wherein the method comprises the following steps: in the step (3), the molar ratio of the rosinyl amine salt to the epichlorohydrin is 1: 1.

5. the method for preparing the environmentally-friendly rosinyl quaternary ammonium salt antibacterial agent according to claim 1, wherein the method comprises the following steps: in the step (3), the antibacterial agent is one of N- (4-abietyloxy-3-hydroxy) butyl-N, N-diethylammonium chloride, N- (3-abietyloxy-2-hydroxy) propyl-N, N-diethylammonium chloride or N- (4-abietyloxy-3-hydroxy) butyl-N, N-dibutylammonium chloride.

Technical Field

The invention relates to the technical field of antibiosis, in particular to a preparation method of an environment-friendly rosin-based quaternary ammonium salt antibacterial agent.

Background

The microorganisms are closely related to the daily life of people, and can bring benefits to people and harm. Most of microorganisms are beneficial to human beings, animals and plants, but some microorganisms can cause diseases to human beings, animals and plants. Contamination of raw materials, production environments and finished products by microorganisms is an important factor causing the defective finished products, and thus the use of antibacterial agents is required to solve the existing problems. Many types of antibacterial agents are sold in the market, but most of the antibacterial agents have serious toxicity and cause serious pollution to the environment. The research and development of new nontoxic or low toxic and high-efficiency antibacterial agents and the exploration of the sterilization effect of the compounds have very important significance.

Disclosure of Invention

In order to solve the problems, the invention provides a preparation method of an environment-friendly rosin-based quaternary ammonium salt antibacterial agent.

In order to achieve the purpose, the technical scheme of the invention is as follows:

the method comprises the following steps:

(1) adding 5-7 parts by weight of rosin and 10-12 parts by weight of epichlorohydrin into a three-neck flask, heating to 60 ℃, dissolving into a homogeneous phase, then dropwise adding 0.15-0.2 part by weight of tetrabutylammonium bromide catalyst, and heating to 95 ℃ for reaction for 5 hours.

After the reaction is finished, the reaction product is rotationally evaporated at 65 ℃ until no distillate exists, and the 3-abietyl oxide-2-hydroxypropyl chloride is obtained as a yellow viscous liquid.

(2) 3-abietyl oxide-2-hydroxypropyl chloride (5-7 parts by weight) and amine salt (7.5-10.5 parts by weight) are used as raw materials, ethanol is used as a solvent, and the reaction is carried out for 8 hours at 95 ℃;

and after the reaction is finished, performing rotary evaporation to remove ethanol and unreacted amine salt to obtain the rosinyl amine salt.

(3) And then adding 5-7 parts by weight of rosinyl amine salt and 5-7 parts by weight of epoxy chloropropane into a three-neck flask, carrying out reflux reaction for 5 hours at 90 ℃, washing with petroleum ether and extracting with ether after the reaction is finished, thus obtaining the rosinyl quaternary ammonium salt antibacterial agent.

Preferably, the amine salt in the step (2) is one of triethylamine, diethylamine or di-n-butylamine;

preferably, the molar ratio of the 3-abietyloxy-2-hydroxypropyl chloride to the amine salt in the step (2) is 1: 1.5;

preferably, the molar ratio of the rosinyl amine salt to the epichlorohydrin in the step (3) is 1: 1;

preferably, the antibacterial agent in the step (3) is one of N- (4-abietyloxy-3-hydroxy) butyl-N, N-diethylammonium chloride, N- (3-abietyloxy-2-hydroxy) propyl-N, N-diethylammonium chloride or N- (4-abietyloxy-3-hydroxy) butyl-N, N-dibutylammonium chloride.

The invention has the beneficial effects that:

(1) the sterilization mechanism of the rosin-based quaternary ammonium salt antibacterial agent is mainly that negative charged bacteria are adsorbed by cations through the actions of electrostatic force, hydrogen bond force, hydrophobic combination between surfactant molecules and protein molecules and the like, and are gathered on cell walls, so that the permeability of the cell walls of the bacteria is changed, a chamber barrier effect is generated, and the bacteria are inhibited from growing and die.

In addition, after the bacterial cell is adsorbed on the surface of a bacterial body, hydrophobic groups (alkyl) and hydrophilic groups (amino) are favorable to respectively penetrate into a lipid layer and a protein layer of the bacterial cell, so that enzyme activity and protein denaturation are caused, a cell structure is damaged, and cell lysis and cell death are caused. Due to the combined effect of the two effects, the rosin-based quaternary ammonium salt antibacterial agent has stronger bactericidal capacity.

(2) The raw materials such as epichlorohydrin and the like selected by the invention have the advantages of low price, rich sources, simple reaction operation steps, high yield of the prepared rosin-based quaternary ammonium salt and excellent antibacterial effect.

(3) On one hand, the rosin-based quaternary ammonium salt prepared by the invention can fully utilize the rosin which is a renewable resource, and the development of the rosin deep processing industry is realized. On the other hand, provides a new path and a method for preparing the antibacterial agent which has low price, high activity, no toxicity, environmental protection and easy biodegradation.

(4) The rosin-based quaternary ammonium salt antibacterial agent prepared by the invention has obvious effects on staphylococcus aureus and escherichia coli, and can realize good antibacterial effect under the condition of low dosage, and the antibacterial rate can reach more than 99%.

Drawings

FIG. 1 is a colony count chart of 24h of cocultivation of different rosin-based quaternary ammonium salt antibacterial agents with Escherichia coli (E bacteria) in the present invention;

FIG. 2 is a colony count chart of 24h of co-culture of different rosin-based quaternary ammonium salt antibacterial agents and Staphylococcus aureus (S bacteria) in the invention;

FIG. 3 shows the results of the bacteriostatic ring test of different rosinyl quaternary ammonium salt antibacterial agents on Escherichia coli (E bacteria) and Staphylococcus aureus (S bacteria) in the invention.

Detailed Description

For further understanding of the present invention, embodiments of the present invention will be described in further detail below with reference to examples and comparative examples, but embodiments of the present invention are not limited thereto.

According to the invention, rosin-based quaternary ammonium salt which is friendly to the environment is synthesized by taking natural product rosin as a raw material, and compared with the influences of different rosin quaternary ammonium salt antibacterial agents on escherichia coli (E bacteria) and staphylococcus aureus (S bacteria), the rosin-based quaternary ammonium salt is found to have a remarkable antibacterial effect, a new path and a new method are provided for the design and preparation of the antibacterial agent, and specific examples are given as follows:

example 1

(1) Adding 5 parts by weight of rosin and 10 parts by weight of epichlorohydrin into a three-neck flask, heating to 60 ℃, dissolving into a homogeneous phase, then dropwise adding 0.15 part by weight of tetrabutylammonium bromide serving as a catalyst, and heating to 95 ℃ for reaction for 5 hours. After the reaction is finished, the reaction product is rotationally evaporated at 65 ℃ until no distillate exists, and the 3-abietyl oxide-2-hydroxypropyl chloride is obtained as a yellow viscous liquid.

(2) 5 parts by weight of 3-abietyloxy-2-hydroxypropyl chloride and 7.5 parts by weight of triethylamine are used as raw materials, ethanol is used as a solvent, the reaction is carried out for 8 hours at 95 ℃, and after the reaction is finished, the ethanol and unreacted triethylamine are removed by rotary evaporation, thus obtaining the N- (4-abietyloxy-3-hydroxy) butyl-N, N-diethylamine.

(3) And then adding 5 parts by weight of N- (4-rosin acyloxy-3-hydroxy) butyl-N, N-diethylamine and 5 parts by weight of epichlorohydrin into a three-neck flask, carrying out reflux reaction for 5 hours at 90 ℃, washing with petroleum ether and extracting with diethyl ether after the reaction is finished, thus obtaining the N- (4-rosin acyloxy-3-hydroxy) butyl-N, N-diethylammonium chloride antibacterial agent A.

Example 2

(1) Adding 6 parts by weight of rosin and 11 parts by weight of epichlorohydrin into a three-neck flask, heating to 60 ℃, dissolving into a homogeneous phase, then dropwise adding 0.2 part by weight of tetrabutylammonium bromide catalyst, and heating to 95 ℃ for reaction for 5 hours. After the reaction is finished, the reaction product is rotationally evaporated at 65 ℃ until no distillate exists, and the 3-abietyl oxide-2-hydroxypropyl chloride is obtained as a yellow viscous liquid.

(2) Taking 6 parts by weight of 3-abietyloxy-2-hydroxypropyl chloride and 9 parts by weight of diethylamine as raw materials, taking ethanol as a solvent, reacting for 10 hours at 50 ℃, and after the reaction is finished, removing the ethanol and unreacted diethylamine by rotary evaporation to obtain the N- (3-abietyloxy-2-hydroxy) propyl-N, N-diethylamine.

(3) And then adding 6 parts by weight of N- (3-rosin acyloxy-2-hydroxy) propyl-N, N-diethylamine and 6 parts by weight of epichlorohydrin into a three-neck flask, carrying out reflux reaction for 5 hours at 90 ℃, washing with petroleum ether and extracting with diethyl ether after the reaction is finished, thus obtaining the N- (3-rosin acyloxy-2-hydroxy) propyl-N, N-diethylammonium chloride antibacterial agent B.

Example 3

(1) Adding 7 parts by weight of rosin and 12 parts by weight of epichlorohydrin into a three-neck flask, heating to 60 ℃, dissolving into a homogeneous phase, then dropwise adding 0.2 part by weight of tetrabutylammonium bromide catalyst, and heating to 95 ℃ for reaction for 5 hours. After the reaction is finished, the reaction product is rotationally evaporated at 65 ℃ until no distillate exists, and the 3-abietyl oxide-2-hydroxypropyl chloride is obtained as a yellow viscous liquid.

(2) Taking 7 parts by weight of 3-abietyloxy-2-hydroxypropyl chloride and 10.5 parts by weight of di-N-butylamine as raw materials, taking ethanol as a solvent, reacting for 8 hours at 95 ℃, and after the reaction is finished, removing the ethanol and the unreacted di-N-butylamine by rotary evaporation to obtain the N- (4-abietyloxy-3-hydroxy) butyl-N, N-dibutylamine.

(3) Then adding 7 parts by weight of N- (4-rosin acyloxy-3-hydroxy) butyl-N, N-dibutylamine and 7 parts by weight of epichlorohydrin into a three-neck flask, carrying out reflux reaction at 90 ℃ for 5 hours, washing with petroleum ether and extracting with diethyl ether after the reaction is finished, thus obtaining the N- (4-rosin acyloxy-3-hydroxy) butyl-N, N-dibutyl ammonium chloride antibacterial agent C.

The test method comprises the following steps:

the bacteriostatic ring method comprises the following steps: adding agar 16.5 weight parts into distilled water 500 weight parts, stirring in water bath (100 deg.C) for 10min, placing into a sterilizing box for 3 hr, pouring under aseptic condition, and cooling.

Bacteria E and S (with a concentration of about 10)⁹CFU/mL) to a bacterial concentration of about 10⁷And (5) uniformly shaking the CFU/mL bacterial solution for 4 hours. Bacteria E and S (with a concentration of about 10)⁷CFU/mL) to a bacterial concentration of about 10⁶Uniformly smearing the CFU/mL bacterial solution on the surface of a nutrient agar medium flat plate (100 muL), waiting for drying, and then pasting three dry sample pieces (each piece is dripped with 20 muL sample solution) and a control sample piece (rosin stock solution) on each flat plate, wherein the total number of the sample pieces is four (the diameter is 5 mm). The sample wafer is lightly pressed with sterile forceps to be tightly attached to the surface of the flat plate. The plate is covered, and the plate is placed in an incubator at 37 ℃, and cultured for 24 hours to observe the result.

Dilution coating plate method: adding agar 16.5 weight parts into distilled water 500 weight parts, stirring in water bath (100 deg.C) for 10min, placing into a sterilizing box for 3 hr, pouring under aseptic condition, and cooling.

Bacteria E and S (with a concentration of about 10)⁹CFU/mL) to a bacterial concentration of about 10⁷And (5) uniformly shaking the CFU/mL bacterial solution for 4 hours. Bacteria E and S (with a concentration of about 10)⁸CFU/mL) to a bacterial concentration of about 10⁷And uniformly shaking the CFU/mL bacterial solution (500 muL bacterial solution +500 muL antibacterial agent +4000 muL LPBS) for 4 h.

Bacteria E and S (with a concentration of about 10) were inoculated using a 24-well plate⁷CFU/mL) to a bacterial concentration of about 10⁶CFU/mL、10⁵CFU/mL、10⁴CFU/mL、10³CFU/mL、10²CFU/mL bacterial liquid, then the bacterial concentration is about 10⁴CFU/mL、10³CFU/mL、10²Respectively coating bacterial liquid of CFU/mL on plates (100 mu L), covering plates, placing in a 37 ℃ incubator, culturing for 24h, and observing knotsAnd (5) fruit.

The results are shown below:

as shown in fig. 1, the culture dish A, B, C treated with the rosin-based quaternary ammonium salt treatment group was said to grow aseptically compared to the control group, and therefore the rosin-based quaternary ammonium salt had a significant effect on escherichia coli (E).

As shown in fig. 2, the rosin-based quaternary ammonium salt-treated petri dish A, B, C was grown aseptically compared to the control group, and thus the rosin-based quaternary ammonium salt was excellent in the anti-staphylococcus aureus (S bacteria) effect.

As shown in FIG. 3 and Table 1, rosin-based quaternary ammonium salt A has the best antibacterial performance against Escherichia coli, and rosin-based quaternary ammonium salt B, C has almost the same antibacterial effect. The rosinyl quaternary ammonium salt C has the best antibacterial performance on staphylococcus aureus (S bacteria), and the rosinyl quaternary ammonium salt C is used for B times. In general, the rosin-based quaternary ammonium salt A, B, C has excellent antibacterial effect.

TABLE 1 different rosin-based quaternary ammonium salt antibacterial ring diameters