阅读说明：本技术 一种高效复合型脂质体消毒剂的制备工艺 (Preparation process of efficient compound liposome disinfectant ) 是由 王海峰 黄海君 王龙 刘星 于 2020-03-09 设计创作，主要内容包括：本发明公开了一种高效复合型脂质体消毒剂的制备工艺,包括将APG、BCDMH、LSM按比例配置成复合药剂,将溶解于碳酸钠/碳酸氢钠溶液中,再与大豆卵磷脂和胆固醇混合,并通过超声乳化处理,形成含脂质体的W/O乳化液；将所述乳化液进行两次减压旋转蒸发,期间需要对所述乳化液进行氮气吹脱盒碳酸钠/碳酸氢钠溶液清洗,获得所述复合型脂质体消毒剂的初级品,经过过滤获得所述复合型脂质体消毒剂的成品。本发明利用脂质体的亲脂功能,运载消毒剂穿透菌胶团/气溶胶的胞外聚合物(EPS)到达微生物内部,增强消毒产品的靶向性,结合绿色表面活性剂APG的渗透通道作用、氧化性含氮消毒剂BCDMH的消毒作用以及LSZ对细胞的生物溶解作用,能够彻底杀灭悬浮型/附着型的微生物。(the invention discloses a preparation process of a high-efficiency compound liposome disinfectant, which comprises the steps of preparing APG, BCDMH and L SM into a compound medicament according to a proportion, dissolving the compound medicament into a sodium carbonate/sodium bicarbonate solution, mixing the compound medicament with soybean lecithin and cholesterol, and performing ultrasonic emulsification treatment to form a W/O emulsion containing liposome, performing reduced pressure rotary evaporation on the emulsion twice, wherein the emulsion needs to be cleaned by a nitrogen blow-off box sodium carbonate/sodium bicarbonate solution during the reduced pressure rotary evaporation to obtain a primary product of the compound liposome disinfectant, and filtering to obtain a finished product of the compound liposome disinfectant.)

1. A preparation process of an efficient compound liposome disinfectant is characterized by comprising the following steps:

S1, preparing APG (alkyl polyglucoside, nonionic surfactant), BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, high-efficiency disinfectant) and L SM (lysozyme ) into a composite medicament in proportion, and dissolving the composite medicament into a sodium carbonate/sodium bicarbonate solution to prepare a composite medicament buffer solution;

S2, dissolving soybean lecithin and cholesterol in ether substances to prepare a first mixed solution;

S3, dissolving the composite medicament buffer solution into the first mixed solution, and performing ultrasonic emulsification treatment to form a liposome-containing W/O emulsion;

S4, carrying out first reduced pressure rotary evaporation on the emulsion, wherein nitrogen stripping is required to be carried out on the emulsion during the first reduced pressure rotary evaporation;

And S5, when a homogeneous film appears on the surface layer of the emulsion, adding a sodium carbonate/sodium bicarbonate solution, and performing secondary reduced pressure rotary evaporation to obtain a primary product of the compound liposome disinfectant.

2. The preparation process of the efficient compound liposome disinfectant as claimed in claim 1, wherein the ether substance is a mixture of chloroform and diethyl ether, and the volume ratio of chloroform to diethyl ether is (1: 3) - (3: 4).

3. The preparation process of the efficient composite liposome disinfectant as claimed in claim 1, wherein the composite liposome disinfectant is filtered by a filtering membrane, so as to obtain a finished product of the composite liposome disinfectant, and the aperture of the filtering membrane is 0.45-0.8 μm.

4. The preparation process of the efficient compound liposome disinfectant as claimed in claim 1, wherein the mass ratio of the soybean lecithin to the cholesterol is (1: 1) - (3: 1).

5. the preparation process of the efficient compound liposome disinfectant as claimed in claim 1, wherein the mass ratio of APG (alkyl polyglucoside, nonionic surfactant), BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, efficient disinfectant) and L YM in the compound medicament is (1: 3: 5) - (1: 10: 10), and the dissolving mass concentration of the compound medicament in sodium carbonate/sodium bicarbonate is 1.5% -4.5%.

6. The preparation process of the efficient compound liposome disinfectant as claimed in claim 1, wherein the volume ratio of the first mixed solution to the compound medicament buffer solution is (1:1) - (1: 5).

7. the preparation process of the efficient compound liposome disinfectant as claimed in claim 1, wherein the concentration of sodium carbonate in a sodium carbonate solution is controlled to be 0.1-0.3 mol/L, and the concentration of sodium bicarbonate in a sodium bicarbonate solution is controlled to be 0.1-0.3 mol/L;

The pH value of the sodium carbonate/sodium bicarbonate solution is controlled to be 9.3-9.86.

8. The preparation process of the efficient compound liposome disinfectant as claimed in claim 1, wherein the reduced-pressure rotary evaporation is performed by using a rotary evaporation device, the working temperature is controlled to be 38-42 ℃, and the working air pressure is controlled to be 0.28-0.38 MPa;

The total time of the first reduced-pressure rotary evaporation and the second reduced-pressure rotary evaporation is 36-64 h.

9. The preparation process of the high-efficiency compound liposome disinfectant as claimed in claim 1, wherein after the reduced-pressure rotary evaporation process, the condensate of chloroform and/or diethyl ether is recycled through a reflux pipeline.

10. The preparation process of the high-efficiency compound liposome disinfectant according to claim 1, wherein the average molecular weight of APG (alkyl polyglucoside, nonionic surfactant) is controlled to be 320-380.

the entrapment rates of the liposome to APG (alkyl polyglucoside, nonionic surfactant), BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, high-efficiency disinfectant) and L SM (lysozyme ) are respectively more than 82%.

Technical Field

The invention relates to the field of killing of microorganisms on the surface of water/air/solid, in particular to a preparation process of an efficient compound liposome disinfectant.

Background

Along with the improvement of the living standard of people, disinfection becomes daily necessary work required by various industries. And the disinfectant has higher removal efficiency requirements for microorganisms in suspension type and attachment type air, otherwise, after the disinfectant is invalid, the microorganisms in the colloid or the zoogloea grow again, and the killing effect of the microorganisms cannot be achieved [ Janus, Liuqian, bamboo stretched in the rain, and the like ] some preliminary recognitions of the novel coronavirus 2019-nCoV [ J ]. environmental chemistry, 2020,39(2): 283-291; influence of Poplar, Below river, alkalinity on removal of biofilms by disinfectants [ J ] chemical management, 2014,11(05): 124-127; research on biofilm optimization control technology on stainless steel surfaces [ D ]. university of congruency, 2009; m, Schneide-Feyrer.S., Huber.C.et al, the interference of custom substituted digital image indications [ J ]. Journal of the Mechanical waveguide of biological Materials 2020,104:660-663 ].

The action principle of the traditional chemical biocides mainly comprises: (1) oxidizing microbial protein amino acids to cause chain scission thereby rendering the protein non-functional, such as chlorine and bromine series biocides; (2) inhibiting the synthesis of microbial enzymes, such as isothiazolinone compounds; (3) cell wall disruption, interaction with cellular proteins, interference with the normal metabolism of cells, such as phenols and quaternary ammonium compounds; (4) adsorbing to the cell wall of the microorganism, diffusing into the cytoplasm, forming a colloidal solution with the cytoplasm, precipitating and denaturing the proteins in the cell, e.g.; a chlorophenolic agent; (5) disrupting microbial energy metabolism processes, such as chlorophenolic agents; (6) adsorbing on the surface of cell membrane and changing its properties, destroying phospholipids in the cell membrane to cause cell autolysis, such as quaternary ammonium compounds; (7) disrupting chloroplasts such as salicylaldehyde and dichlorophen in algal cells; (8) affecting the growth and division of bacterial cells, spore germination and respiration, inhibiting cell expansion, and disintegrating cytoplasm, such as alkylguanidine; (9) heavy metal ions bind to cellular proteins or enzymes to inactivate them, such as heavy metal salts; (10) adsorption with negatively charged microorganisms occurs, and long chains are wound around the microorganisms to hinder the gene expression process, such as quaternary phosphonium salts. However, microorganisms have proven to be the most environmentally compatible and effective drug-resistant microorganisms on earth. The microorganisms protect their own survival by secreting extracellular polymers and group gene replication, and form polymers of other microorganisms, preventing the effective entry of disinfectants.

The surfactant is a substance having both a hydrophilic group and a hydrophobic group in a molecule. Such substances can significantly alter the tension at the interface of the two phases, thereby facilitating dispersion of the pharmaceutical agent in aqueous solution. The disadvantages are that: the single use of the composition has low efficiency of killing microorganisms and is greatly influenced by pH.

The enzyme is protein with special structure and function, and has the characteristics of high efficiency, specificity and the like. The enzyme can effectively attack mucous membrane layers around bacteria, so that microorganisms are dispersed, a biological membrane is not formed any more, and the removal capacity of the disinfectant can be enhanced [ slow, Wanghang peak, plum aegiline ] lysozyme liposome preparation and the stripping effect on the biological membrane [ J ]. Tongji university newspaper (Nature science edition).: 2011,39(1):90-94 ].

the Chinese invention patent Z L201110181901.1 describes a technique for controlling a pipeline biofilm by using a bromochlorohydantoin liposome, and only BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, a high-efficiency disinfectant) liposome is used for analyzing the disinfection effect, which belongs to an oxidative disinfectant and has a good effect on Escherichia coli.

Japanese patent JP 2008-63304 discloses a disinfectant based on quaternary ammonium salt, which has only high killing efficiency on microorganisms in water, and lacks deep discussion on killing aerosol and zoogloea microorganisms.

The chinese invention patent CN201911041301.8 discloses a method for disinfection in a pipeline, which only uses a conventional chlorine-containing disinfectant to clean the pipeline, and does not select the disinfectant according to the property of the pipeline enriched with microbial micelles, and the treatment efficiency is often low in the actual process.

Before 150 years, humans found: traditional microbial growth favors surface growth. For example, the amount of microorganisms in the pipeline that are enriched on the surface of the pipeline often accounts for more than 90%, and the removal of microorganisms from the water body cannot effectively control the total amount of microorganisms in the whole pipeline system. Microorganisms, particularly viruses, in the air are also present on the surface of the aerosol (human droplets also belong to the aerosol), and the microorganisms attached to the surface of the aerosol can perform complete gene expression through the attached microorganisms through the population effect ("quorum-sense"), so that the attached microorganisms can have the gene segments which are most suitable for survival.

The adhered bacteria undergo new metabolism and growth to form colonies (Community) containing various microorganisms, the microorganisms in the colonies form a protective layer having mechanical strength by secreting various Extracellular Polymers (EPS), and finally form a biofilm structure having a stable structure by continuous new metabolism, shedding and re-growth, resulting in poor treatment efficiency of the conventional disinfectant. At present, the virus microorganisms with high mutation probability and strong replication capacity are often present in a 'host', and are expressed through complete genes. How to break the zoogloea protective layer efficiently to form sterilization, permeation, cell wall dissolution and coexistence of hydrophilicity and hydrophobicity, the development of a targeted compound disinfectant is not found, and the processing capacity similar to that of a targeted drug is achieved through sustained release.

Disclosure of Invention

aiming at the defects and shortcomings of the prior art, the preparation process of the efficient compound liposome disinfectant comprises the following steps of S1, preparing APG (alkyl polyglucoside and nonionic surfactant), BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, efficient disinfectant) and L SM (lysozyme ) into compound medicaments according to a proportion, dissolving the compound medicaments into a sodium carbonate/sodium bicarbonate solution to prepare a compound medicament buffer solution, S2, dissolving soybean lecithin and cholesterol into ether substances to prepare a first mixed solution, S3, dissolving the compound medicament buffer solution into the first mixed solution, performing ultrasonic emulsification treatment to form a liposome-containing W/O emulsion, S4, performing first reduced pressure rotary evaporation on the emulsion, blowing off nitrogen gas to the emulsion during the first reduced pressure rotary evaporation, and S5, adding the surface layer/sodium carbonate solution into the emulsion after the emulsion appears a homogeneous film, and performing second reduced pressure rotary evaporation on the emulsion to obtain a compound sodium carbonate solution.

Preferably, the ether substance is a mixture of chloroform and diethyl ether, and the volume ratio of chloroform to diethyl ether is (1: 3) - (3: 4).

Preferably, the compound liposome disinfectant is filtered by a filter membrane to obtain a finished product of the compound liposome disinfectant, wherein the aperture of the filter membrane is 0.45-0.8 μm.

Preferably, the mass ratio of the soybean lecithin to the cholesterol is (1: 1) - (3: 1).

preferably, the mass ratio of APG (alkyl polyglucoside, nonionic surfactant), BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, high-efficiency disinfectant) and L YM in the compound medicament is (1: 3: 5) - (1: 10: 10), and the dissolving mass concentration of the compound medicament in sodium carbonate/sodium bicarbonate is 1.5% -4.5%.

Preferably, the volume ratio of the first mixed solution to the composite medicament buffer solution is (1:1) - (1: 5).

preferably, the concentration of sodium carbonate in the sodium carbonate solution is controlled to be 0.1-0.3 mol/L, the concentration of sodium bicarbonate in the sodium bicarbonate solution is controlled to be 0.1-0.3 mol/L, and the pH value of the sodium carbonate/sodium bicarbonate solution is controlled to be 9.3-9.86.

Preferably, a rotary evaporation device is adopted for carrying out the reduced pressure rotary evaporation, the working temperature is controlled to be 38-42 ℃, and the working air pressure is controlled to be 0.28-0.38 MPa; the total time of the first reduced-pressure rotary evaporation and the second reduced-pressure rotary evaporation is 36-64 h.

Preferably, after the reduced pressure rotary evaporation process, the condensate of chloroform and/or diethyl ether is recovered for reuse through a reflux pipeline.

preferably, the average molecular weight of APG (alkyl polyglucoside, nonionic surfactant) is controlled to be 320-380, and the encapsulation rates of the liposome to APG (alkyl polyglucoside, nonionic surfactant), BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, high-efficiency disinfectant) and L SM (lysozyme ) are respectively more than 82%.

BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, high-efficiency disinfectant) belongs to a disinfectant with specific performance and has the function of slow-release disinfection

Due to the adoption of the technical scheme, compared with the prior art, the invention has the following advantages and positive effects:

1. the invention provides a preparation process of a high-efficiency compound liposome disinfectant, which utilizes the lipophilic function of liposome to carry a disinfectant to penetrate Extracellular Polymer (EPS) of zoogloea/aerosol to reach the inside of microorganisms, enhances the targeting of a disinfection product, combines the permeation channel effect of a green surfactant APG (alkyl polyglucoside, nonionic surfactant), the disinfection importance of an oxidative nitrogenous disinfectant BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, high-efficiency disinfectant) and the biological dissolution effect of L SZ (lysozyme ) on cells, has multiple technical effects of slow release, health of human bodies, cell wall dissolution and hydroxyl radical disinfection, has high-efficiency killing effects on suspended and attached microorganisms in water bodies/gases, has wide office application, relatively low efficient killing efficiency on zoogloom or relatively low efficient killing efficiency compared with the zoogloom disinfectant, can achieve the high-efficiency killing effect of removing suspended and balanced hydrophilic zooglomus/aerosol, can meet the environment-friendly disinfection scenes of environmental protection and environmental protection.

2. The preparation process of the high-efficiency compound liposome disinfectant provided by the invention can blow off oxygen in the rotary evaporation device by nitrogen blowing off, and reduce the oxidation water in synthesis.

3. According to the preparation process of the efficient compound type liposome disinfectant, provided by the invention, the sodium carbonate/sodium bicarbonate solution is added in the decompression rotary evaporation process, so that the solution in the liposome is in a stable state, and the liposome is also in a stable state system.

Drawings

FIG. 1 is a process flow diagram of the present invention;

Fig. 2 is a scanning electron microscope SEM image of the complex pharmaceutical agent liposome of the present invention;

Fig. 3 is a TEM image of a projection electron microscope of a complexed pharmaceutical agent liposome of the present invention;

FIG. 4 is a table showing Zeta potential of complex liposomes under ultrasound conditions according to the present invention as a function of time.

Detailed Description

The invention will be described in more detail hereinafter with reference to the accompanying drawings, in which embodiments of the invention are shown. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. In the drawings, the size and relative sizes of layers and regions may be exaggerated for clarity.

As shown in figure 1, the invention provides a preparation process of a high-efficiency compound type liposome disinfectant, which comprises the steps of preparing APG (alkyl polyglucoside, nonionic surfactant), BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, high-efficiency disinfectant) and L SM (lysozyme ) into compound medicaments according to a proportion, dissolving the compound medicaments into a sodium carbonate/sodium bicarbonate solution to prepare a compound medicament buffer solution, dissolving soybean lecithin and cholesterol into ether substances according to the proportion of (1: 1) to (3: 1) to prepare a first mixed solution, wherein the ether substances are usually a mixture of chloroform and diethyl ether, the volume ratio of the chloroform to the diethyl ether can be replaced by dichloromethane or other ethers with the effect of not better than that of the chloroform and the diethyl ether, dissolving the compound medicament buffer solution into the first mixed solution, performing emulsification ultrasonic treatment to form a W/O emulsion containing liposome, performing rotary evaporation on the first-reduced-pressure emulsion, performing rotary evaporation on the compound type liposome disinfectant during the first-reduced-pressure evaporation, and the first-reduced-concentration of the compound type liposome disinfectant is reduced by blowing nitrogen, and the compound type sodium bicarbonate solution is obtained after the compound type liposome emulsion is cooled, and the compound type sodium bicarbonate solution is cooled, and the concentration of the compound type sodium carbonate solution is reduced by adding a sodium carbonate/sodium carbonate solution, wherein the compound type sodium carbonate solution is reduced by adding a sodium carbonate solution, and adding a sodium carbonate solution is reduced by adding a sodium carbonate solution, and adding a sodium carbonate solution, wherein the compound type sodium carbonate solution is reduced by adding a sodium carbonate solution.

The efficient compound liposome disinfectant disclosed by the invention is simple to prepare, strong in disinfection spectrum, capable of being stored for a long time, high in removal efficiency of viruses (including coronavirus) in aerosol, and good in removal effect on gram-positive bacteria and gram-negative bacteria. The process flow of the invention has strong adaptability, can be suitable for the preparation of liposome of disinfectants with different scales and different types, has high degree of process automation, and can conduct dynamic data analysis by 5G and other technologies so as to guide the operation.

furthermore, the mass ratio of APG (alkyl polyglucoside, nonionic surfactant), BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, high-efficiency disinfectant) and L YM in the composite medicament is (1: 3: 5) - (1: 10: 10), the average molecular weight of APG (alkyl polyglucoside, nonionic surfactant) is controlled to be 320-380, the encapsulation rates of liposome to APG (alkyl polyglucoside, nonionic surfactant), BCDMH (bromochlorohydantoin, 1-bromo-3-chloro-5, 5-dimethylhydantoin, high-efficiency disinfectant) and L SM (lysozyme, L OZYME) are respectively greater than 82%, the dissolving mass concentration of the composite medicament in sodium carbonate/sodium bicarbonate is 1.5% -4.5%, and the volume ratio of the first mixed solution to the composite medicament buffer solution is (1: 5).

furthermore, if a sodium carbonate solution is used, the concentration of sodium carbonate in the sodium carbonate solution is controlled to be 0.1-0.3 mol/L, if a sodium bicarbonate solution is used, the concentration of sodium bicarbonate in the sodium bicarbonate solution is controlled to be 0.1-0.3 mol/L, and the pH value of the sodium carbonate/sodium bicarbonate solution is adjusted to be 9.3-9.86 because the compound type liposome disinfectant is an organic matter, so that alkalescence is kept, and the organic matter can be prevented from being oxidized or other processes.

Further, a rotary evaporation device is adopted for carrying out reduced pressure rotary evaporation, the working temperature is controlled to be 38-42 ℃, and the working air pressure is controlled to be 0.28-0.38 MPa; the total time of the first reduced-pressure rotary evaporation and the second reduced-pressure rotary evaporation is 36-64 h. After the decompression rotary evaporation process, the condensate of the chloroform and/or the diethyl ether is recycled through a reflux pipeline, the use of the solvent is reduced, and secondary environmental pollutants are not generated.

The present invention will be described in detail with reference to embodiments for different application scenarios and different process parameters.