阅读说明：本技术 十二烷基二甲基羟丙基三甲基氯化二铵及其合成方法与应用 (Dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride and synthetic method and application thereof ) 是由 刘晓林 段梦婕 段秀鹏 于 2021-10-22 设计创作，主要内容包括：本发明公开了十二烷基二甲基羟丙基三甲基氯化二铵及其制备方法与应用。十二烷基二甲基羟丙基三甲基氯化二铵,其理化性质如下：性状：黄白色蜡状固体或胶状体；熔点50-55℃；溶解性：易溶。药效学结果表明,按1∶600使用本发明提供的十二烷基二甲基羟丙基三甲基氯化二铵能有效杀灭猪舍环境中的金黄色葡萄球菌、大肠埃希氏杆菌和溶血性链球菌等,杀菌率达91.9％。十二烷基二甲基羟丙基三甲基氯化二铵可用于环境消毒、厩舍、饲喂器具、畜禽、种蛋、饮水的消毒灭菌等。(The invention discloses dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride and a preparation method and application thereof. The physical and chemical properties of the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride are as follows: the characteristics are as follows: an off-white waxy solid or gel; melting point is 50-55 ℃; solubility: is easy to dissolve. Pharmacodynamic results show that the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride provided by the invention can effectively kill staphylococcus aureus, escherichia coli, hemolytic streptococcus and the like in a pigsty environment according to the ratio of 1: 600, and the sterilization rate reaches 91.9%. The dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride can be used for environmental disinfection, barnyard houses, feeding appliances, livestock and poultry, hatching eggs, drinking water disinfection and the like.)

1. A compound having the structural formula I:

(formula I).

2. A process for the preparation of a compound of formula I according to claim 1, comprising the steps of:

under the condition of stirring, carrying out primary quaternization reaction on dodecyl dimethylamine and epoxy chloropropane, stopping stirring after the reaction is finished, adding trimethylamine hydrochloride into the system to carry out secondary quaternization reaction, and finishing the reaction to obtain the compound shown in the formula I.

3. The method of claim 2, wherein: the reaction temperature of the first quaternization reaction is controlled to be 80-110 ℃, and the reaction time is 4 hours;

the reaction temperature of the second quaternization reaction is controlled to be 80-110 ℃, and the reaction time is 3 hours.

4. The production method according to claim 2 or 3, characterized in that:

the mass ratio of the dodecyl dimethylamine to the epichlorohydrin is (400-) -500): 241, a first electrode and a second electrode;

the mass ratio of the trimethylamine hydrochloride to the epichlorohydrin is (180-) -220): 241.

5. the use of a compound of formula I as claimed in claim 1, comprising at least one of:

1) the application in the preparation of disinfectants;

2) the application in the preparation of sterilizing agents;

3) application in preparing antiseptic.

6. Use according to claim 5, characterized in that: the sterilizing agent can kill gram-positive bacteria and gram-negative bacteria, specifically staphylococcus aureus, escherichia coli and hemolytic streptococcus.

7. Use according to claim 5, characterized in that: the disinfectant and the sterilizing agent are used for sterilizing the environment, barnyards, feeding appliances, livestock and poultry, hatching eggs and drinking water.

8. A method of disinfecting an environment or stall, comprising the steps of: the compound solution of formula I according to claim 1 diluted with water is sprayed into the environment or stable.

9. The method of claim 8, wherein: the mass concentration of the compound solution shown in the formula I is 45%; the mass ratio of the compound solution of the formula I to water is 1: 600.

Technical Field

The invention belongs to the field of chemical synthesis, and particularly relates to dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride as well as a synthesis method and application thereof.

Background

The quaternary ammonium salt is one of the bactericides which are widely applied in the future, and the disinfectant has the characteristics of low toxicity, safety, small side effect, low concentration, effectiveness, no color, no odor and low irritation. Since the 50 s of the last century, quaternary ammonium salt disinfectants developed to date, hundreds of varieties were answered. According to their structure, they can be divided into four classes, single-chain quaternary ammonium salts, double-chain quaternary ammonium salts, polyquaternary ammonium salts. The double-chain quaternary ammonium salt is represented by didecyl dimethyl ammonium chloride (thalidomide), didecyl dimethyl ammonium bromide (chlorhexadione), didodecyl dimethyl ethylene diammonium bromide (neojieling) and the like, and can kill various microorganisms including bacteria, algae and viruses. But the bactericidal performance of the quaternary ammonium salt is closely related to the molecular structural formula, so that the development of the quaternary ammonium salt product with more excellent bactericidal performance has important practical significance.

Disclosure of Invention

The invention aims to provide dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride and a preparation method thereof.

The structural formula of the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride provided by the invention is shown as a formula I:

(formula I)

The molecular formula of the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride is C₂₁H₄₈N₂OCl₂Molecular weight of

415.53. The physical and chemical properties are as follows: the characteristics are as follows: an off-white waxy solid or gel; melting point is 50-55 ℃; solubility: is easy to dissolve.

The preparation method of the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride provided by the invention comprises the following steps:

under the condition of stirring, carrying out primary quaternization reaction on dodecyl dimethylamine and epoxy chloropropane, stopping stirring after the reaction is finished, adding trimethylamine hydrochloride into the system to carry out secondary quaternization reaction, and finishing the reaction to obtain the target product dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride.

The reaction temperature of the first quaternization reaction is controlled to be 80-110 ℃, and the reaction time is 4 hours.

The reaction temperature of the second quaternization reaction is controlled to be 80-110 ℃, and the reaction time is 3 hours.

In the first quaternization reaction, the mass ratio of the dodecyl dimethyl amine to the epichlorohydrin is (400-) -500): 241, specifically 453: 241.

in the second quaternization reaction, the mass ratio of the trimethylamine hydrochloride to the epichlorohydrin is (180-) -220): 241, specifically, 195: 241.

the invention also provides application of the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride.

The application of the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride provided by the invention comprises at least one of the following components:

1) the application in the preparation of disinfectants;

2) the application in the preparation of sterilizing agents;

3) application in preparing antiseptic.

The sterilizing agent can kill gram-positive bacteria and gram-negative bacteria, and particularly can kill staphylococcus aureus, escherichia coli and hemolytic streptococcus.

The disinfectant and the sterilizing agent can be used for environmental disinfection, stable houses, feeding appliances, livestock and poultry, hatching eggs, drinking water disinfection and the like.

The invention also protects a method for disinfecting an environment or a stable.

The method for disinfecting the environment or the stable provided by the invention comprises the following steps: the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride is diluted by water and then sprayed into the environment or a stable house.

The stable can be sprayed and then the door and the window can be closed for 1h, so that the disinfection effect is improved.

In the method, the mass ratio of the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride to water can be 1: 300-900, the preferred mass ratio is 1: 300-600.

Pharmacodynamic results show that the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride provided by the invention can effectively kill staphylococcus aureus, escherichia coli, hemolytic streptococcus and the like in a pigsty environment according to the ratio of 1: 600, and the sterilization rate reaches 98.9%.

Drawings

FIG. 1 is a flow chart of a production process of dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride.

FIG. 2 is an infrared identification spectrum of dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride.

Detailed Description

The present invention will be further illustrated with reference to the following specific examples, but the present invention is not limited to the following examples. The method is a conventional method unless otherwise specified. The starting materials are commercially available from the open literature unless otherwise specified.

The starting materials used in the following examples are as follows:

1. trimethylamine hydrochloride, content limit is not less than 99.0%, supplier: shandonglidingxin section Material Co., Ltd;

2. dodecyl dimethylamine, limit of content ≥ 97.0%, supplier: clarienfengyi fatty amines (lianyuankang) limited;

3. epichlorohydrin, the content limit is more than or equal to 99.0%, and the supplier: shandong Xin Chemicals, Inc.

In the following examples, the quaternization reaction process is carried out in a stainless steel electric heating porcelain kettle; the filtration procedure was carried out in a precision filter, equipment type RG-6.

Example 1 Synthesis of dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride

The flow chart of the production process of dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride is shown in figure 1.

Firstly, adding weighed tertiary amine (dodecyl dimethyl amine) (453 g) into an enamel kettle, starting stirring, stopping stirring and adding epichlorohydrin (241 g) after 30 minutes, starting quaternization, starting to increase the temperature of materials from room temperature, starting to introduce cooling water when the temperature exceeds 80 ℃, and controlling the temperature not to exceed 110 ℃. Keeping the temperature of 80-110 ℃ (100 ℃ is selected specifically) for continuous reaction for 4 hours, stopping stirring, adding trimethylamine hydrochloride (195 g) to start the second quaternization reaction, and keeping the temperature of 80-110 ℃ (100 ℃ is selected specifically) for continuous reaction for 3 hours to obtain the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride (the yield is 90%, and the purity is 95%).

The infrared spectrum is shown in FIG. 2.

Through structure identification, the obtained compound is the target product dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride.

The key process parameters of the reaction are as follows: the reaction temperature is controlled to be 80-110 ℃, and the reaction time is not less than 7 hours

Detecting a target product:

1. the characteristics are as follows: the target product should be yellow colloid at normal temperature, and wax-like solid can be gradually formed at low temperature; the aqueous solution shows alkaline reaction and generates a large amount of foam when shaking;

2. non-quaternary ammonium species: the difference between potassium iodate titration solution (0.05 mol/L) consumed in two times is not more than 0.5 ml;

3. content determination:

the content was determined by potentiometric titration. Calculated as anhydrate, the ammonium salt containing hydrocarbons (C)₂₀H₄₆N₂OCl₂) 97.0% -103.0%.

And opening a discharge port of the reaction kettle after the semi-finished product is qualified, putting the semi-finished product into a storage barrel, starting a filter, and carrying out filtering operation. And then pumping the filtered product into a target packaging barrel, and taking the label, wherein the batch number, the production date and the effective period on the label are printed.

Comparative example 1: synthesis of dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride

Firstly adding weighed tertiary amine (dodecyl dimethyl amine) (453 g) into an enamel kettle, starting stirring, stopping stirring and adding epichlorohydrin (241 g) after 30 minutes, starting quaternization, starting temperature rise from room temperature, keeping the temperature for continuous reaction for 4 hours when the temperature reaches 60-70 ℃ (specifically 65 ℃), stopping stirring, adding trimethylamine hydrochloride (195 g) to start second quaternization, keeping the temperature at 60-70 ℃ (specifically 65 ℃) for continuous reaction for 3 hours, and obtaining dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride (yield is 60%, purity is 80%).

Comparative example 2: synthesis of dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride

Firstly, adding weighed tertiary amine (dodecyl dimethyl amine) (453 g) into an enamel kettle, starting stirring, stopping stirring and adding epichlorohydrin (241 g) after 30 minutes, starting quaternization, starting to increase the temperature of materials from room temperature, starting to introduce cooling water when the temperature exceeds 80 ℃, and controlling the temperature not to exceed 110 ℃. Keeping the temperature of 80-110 ℃ for continuous reaction for 3 hours, stopping stirring, adding trimethylamine hydrochloride (195 g) to start secondary quaternization, keeping the temperature of 80-110 ℃ for continuous reaction for 2.5 hours to obtain the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride (the yield is 75%, and the purity is 86%).

Example 2 efficacy test of dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride

The disinfection work is one of the important links for preventing and controlling the occurrence of epidemic diseases, and is a main means for cutting off the propagation of pathogenic microorganisms. In recent years, high-efficiency, quick and broad-spectrum disinfectants are sought at home and abroad. The dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride prepared by the invention has a killing effect on staphylococcus aureus, escherichia coli and hemolytic streptococcus C groups in the environment. Laboratory tests for bacteriostasis and piggery environment disinfection were performed to confirm the disinfection effect, and are reported below.

Materials and methods

1. Test materials

1.1 test species Staphylococcus aureus, Escherichia coli, and Streptococcus hemolyticus group C, each represented by A, B, C. Provided by the subject group.

1.2 reagent 10g/L phosphate buffer solution of peptone, common nutrient agar culture medium calf serum, Martin broth culture medium

1.3 degreasing white plain cloth (10 mm. times.10 mm), PQ-240 type spray sterilizer.

1.4 pigsty for experiment: provided by the pig farm of the livestock veterinary research institute of shanxi academy of agricultural sciences.

1.5 medicine: dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride: manufactured by Shanxi Nuo Reed Biotechnology Co., Ltd (prepared according to the method of example 1), neutralizing agents glycine and Tween-80 were purchased from the market. Dodecyl dimethyl benzyl ammonium chloride disinfectant: shandongtai and water treatment, batch number: 200716, specification: 40 percent.

2. Test method

2.1 neutralizing agent Glycine and Tween-80 were selected as neutralizing agents for disinfectants according to reference [1], and the optimum neutralizing concentrations were determined.

2.2 bacterial liquid preparation transfer the bacteria to 6 th-8 th generation, and dilute the bacteria into 5X 10 with PBS⁶CFU/mL bacterial suspension.

2.3 Sterilization test

2.3.1 qualitative Sterilization test

Diluting dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride disinfectant with sterilized distilled water to different concentrations, adding an equal amount of test bacterium liquid into 2.5mL of each tube, mixing uniformly, placing in a water bath at 20 ℃, taking 0.5mL of each tube every 5, 10, 15, 30 and 60min, adding into 4.5mL of Martin broth containing a neutralizing agent, shaking uniformly, neutralizing for 10min, taking out 0.5mL of each tube, adding into 4.5mL of Martin broth culture medium, culturing at 37 ℃ for 24h, observing the growth condition of bacteria, showing the growth of bacteria by turbidity, continuing to culture until the 7 th day without turbidity, and judging the bacteria can not grow aseptically. Meanwhile, a control tube is arranged.

2.3.2 quantitative Sterilization test with Carrier

Escherichia coli and Staphylococcus aureus were respectively made into bacterial suspensions in Phosphate Buffered Saline (PBS) containing 10g/L peptone. 0.02ml of the bacterial suspension was applied and dropped on a piece of defatted white plain cloth (10 mm. times.10 mm). Drying in 37 deg.C incubator for 20min to obtain bacterial tablet. For the experiment, 1 bacterial tablet was added to 5.0ml of disinfectant (PBS as positive control). Acting for a prescribed time, taking out the bacterial tablet, transferring into a test tube containing 50ml of neutralizer solution, neutralizing for 10min, beating the test tube to elute bacteria on the bacterial tablet, and mixing uniformly. 0.5ml of eluent is taken as common nutrient agar for pouring inoculation, and the mixture is placed at 37 ℃ for culture for 48 hours, and the result is observed. And calculating the killing rate.

2.3.3 when observing the influence of organic matter, the bacterial suspension is added with calf serum and then infected with bacterial slices, and the experiment is carried out according to the procedure.

2.3.4 piggery pigsty disinfection test

The pigsty is selected for observation of on-site disinfection effect, the pigsty is of a brick-concrete structure, the metal fence is used for feeding, and the cement ground is used. After cleaning and washing with tap water, drying, using a PQ-240 type spray disinfection machine to divide the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride disinfectant into test groups 1, 2 and 3, and spray the diluted solutions according to the ratio of 1:300, 1: 600 and 1: 900, wherein 4 groups are control groups (dodecyl dimethyl benzyl ammonium chloride), and the doors and windows are closed for 1 hour after spraying. Placing agar culture medium plates at four corners and five points in the middle of a pigsty by a plate exposure method respectively before and after disinfection, and placing for about half an hour for sampling; and then placing the agar culture medium plate into an incubator at 37 ℃ for culturing for 24h, counting the number of bacterial colonies before and after disinfection, and calculating the killing rate.

3. Observation items

3.1 growth of bacteria in Martin broth, positive for turbidity and negative for no turbidity.

3.2 variation in the number of colonies grown by bacteria on agar medium before and after sterilization.

Second, test results

1. Effect of neutralizing agent

The result shows that the staphylococcus aureus is taken as an indicator bacterium: the buffer solution of 50g/L glycine and Tween-80 is used as a neutralizer to effectively neutralize 10% disinfectant solution, and does not affect the growth of bacteria.

2. Sterilizing effect

2.1 the three kinds of bacteria respectively act with disinfectants with different concentrations for different time, the lowest concentration of the disinfectants of the aseptic growth tube is taken as the lowest effective sterilization concentration, and the shortest sterilization time of the aseptic growth tube is taken as the fastest effective sterilization time of the concentration (the result is shown in table 1).

TABLE 1 qualitative bactericidal test results of dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride disinfectant on bacteria

Note that A, B, C represents Staphylococcus aureus, Escherichia coli, and hemolytic streptococcus, respectively;

② "+" indicates the growth of bacteria, "-" indicates the growth of no bacteria;

results in the table are the average of 6 tests.

As can be seen from Table 1, the lauryl dimethyl hydroxypropyl trimethyl diammonium chloride disinfectant can effectively kill staphylococcus aureus, Escherichia coli and hemolytic streptococcus within 10min when diluted by 1: 700.

2.2 quantitative germicidal test with vehicle (results in Table 2)

TABLE 2 list of the effect of sterilization at different times

Note: (1) the test temperature is 19-22 ℃;

(2) the number of the positive control group escherichia coli is 1310000 cfu/tablet;

(3) the number of staphylococcus aureus is 1500000 cfu/tablet;

(4) the results are the average of 3 trials.

As can be seen from table 2: the disinfection effect of staphylococcus aureus and escherichia coli for 7min can reach the killing rate of more than 99.10%.

2.3 organic matter Effect test

When the bacterial suspension contained 50% by volume of calf serum, the killing effect of staphylococcus aureus was slightly affected by the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride disinfectant (the results are shown in table 3).

Table 3 organic matter influence test results

2.4 piggery pigsty disinfecting effect

Disinfection of piggery with dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride disinfectant (results are shown in Table 4)

TABLE 4 bacterial colony count before and after piggery and pigsty disinfection

As can be seen from table 4: after the disinfection by the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride, the colony number of the pigsty is obviously reduced, and the sterilization rates of the test groups 1, 2 and 3 and the control group are respectively 99.5%, 98.9%, 91.5% and 91.1%.

III, III and discussion

1. From the aspect of the effect of killing bacteria, the preparation fully exerts the effective killing effect, expands the bactericidal spectrum and is convenient to use. Dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride destroys protein receptors on the surface of bacteria. Under the condition of a laboratory, even if the diluted solution is diluted by 1: 700 times, the staphylococcus aureus, the escherichia coli and the hemolytic streptococcus can be effectively killed within 10 min; can kill gram-positive bacteria and gram-negative bacteria, and has a sterilization effect of staphylococcus aureus and escherichia coli of over 99.10 percent in 7min in a carrier quantitative sterilization test. When the bacterial suspension contains calf serum with the volume fraction of 50%, the dodecyl dimethyl hydroxypropyl trimethyl diammonium chloride disinfectant has a slight influence on the killing effect of staphylococcus aureus.

2. On-site sterilization, test components are 1:300, 1: 600. 1: three doses of 900, the sterilization rate at 60min is 99.5%, 98.9% and 91.5%, respectively. Control group 1: 600, the sterilization rate is 91.1%. Therefore, the recommended dose for clinical use is: 1: 600.

3. When the disinfection effect of the disinfectant is researched under the laboratory condition, when the mixed solution is transferred into the culture medium, a small amount of residual disinfectant is inevitably carried, the growth and the propagation of bacteria in the culture medium can be continuously hindered,form a negative result^[2]Therefore, the effect of residual disinfectant must be removed in the bactericidal concentration test. In the present study, the selected neutralizing agent can neutralize the disinfectant well, and the concentration of the neutralizing agent does not affect the growth of bacteria.

Reference documents:

[1] fouqiang and Liu Zhan military poultry farm poultry disease inspection manual [ M ]: beijing: the Chinese agriculture university Press, 1992.130-133.

[2] The iodine-containing cleaning disinfectant has the following effects on killing clinical microorganisms when being erected, grandfather, Yanxuming and the like [ J ]: journal of china disinfection, 1998, 15 (2): 108 to 110.