CN111423851B

CN111423851B - Biological aerosol simulator and preparation method thereof

Info

Publication number: CN111423851B
Application number: CN202010097352.9A
Authority: CN
Inventors: 粟勇; 袁丁; 吴红彦; 夏征
Original assignee: Ht Nova Co ltd
Current assignee: Ht Nova Co ltd
Priority date: 2020-02-17
Filing date: 2020-02-17
Publication date: 2023-09-08
Anticipated expiration: 2040-02-17
Also published as: CN111423851A

Abstract

The application relates to the technical field of biological aerosol, in particular to a biological aerosol simulator and a preparation method thereof. The biological aerosol simulator with flexible control of shape, low cost and good fluorescent effect is provided by adopting the mode of mixing the fluorescent agent and the surfactant.

Description

Biological aerosol simulator and preparation method thereof

Technical Field

The application relates to the technical field of biological aerosol, in particular to a biological aerosol simulator and a preparation method thereof.

Background

Bioaerosols refer to aerosols containing biological particles. Including bacteria, viruses, sensitized pollen, mould spores, fern spores, parasitic ova and the like, and has infectivity, sensitization and the like in addition to the characteristic of common aerosol.

When the bioaerosol is irradiated with light of a certain wavelength, a specific fluorescence is emitted, so that the bioaerosol alarm can be prepared according to the principle. For such alarms, aerosols with fluorescent properties can be used as simulators for earlier investigation and demonstration of the device. At present, fluorescent polymer microspheres are used as a simulator, and an aqueous solution of the fluorescent polymer microspheres is atomized to generate aerosol for simulation test. Alternatively, aqueous riboflavin may be used as a simulator.

The method for simulating the biological aerosol by adopting the fluorescent microspheres is very limited in shape, the obtained aerosol is mostly spherical, other biological aerosols except the spherical cannot be truly simulated, and the fluorescent polymer microspheres have higher manufacturing cost and are inconvenient to use in a large amount. For the method using riboflavin, the fluorescence is relatively weak, and the actual test effect is poor.

Therefore, there is an urgent need in the market for bioaerosol simulators that can be flexibly controlled in shape, are low in cost, and have good fluorescence effects.

Disclosure of Invention

The application aims to solve the problems and the disadvantages, and provides a bioaerosol simulator which has flexible control of shape, low cost and good fluorescence effect by adopting a mixture of a fluorescent agent and a surfactant.

According to one aspect of the present application, there is provided a bioaerosol simulation agent comprising a fluorescent agent and a surfactant.

Wherein the fluorescent agent is selected from one or more of naphthalene, anthracene, phenanthrene and derivatives thereof, and heterocyclic compounds with fluorescent signals.

Wherein the surfactant is selected from cationic surfactant, anionic surfactant and nonionic surfactant.

Preferably, the surfactant is selected from quaternary ammonium salts, carboxylates, alkyl sulfonates, alkylbenzene sulfonates or polysorbates.

More preferably, the surfactant is selected from cetyltrimethylammonium bromide (CTAB), cetyltrimethylammonium chloride (CTAC), dodecyltrimethylammonium bromide (DTAB), dodecyltrimethylammonium chloride (DTAC), sodium Dodecyl Sulfate (SDS), sodium Tetradecyl Sulfate (STS), sodium hexadecyl sulfate, sodium octadecyl sulfate, potassium stearate, potassium oleate Sodium Dodecyl Benzene Sulfonate (SDBS), or tween 80.

The fluorescent agent, also called fluorescent brightening agent, is a fluorescent dye or white dye, and is characterized by that it can absorb incident light ray to produce fluorescence so as to make the dyed material obtain the effect of fluorspar-like flash luminescence, and make the material seen by naked eye be very white so as to attain the effect of whitening. The effect is that the invisible ultraviolet radiation absorbed by the product is converted into purple blue fluorescent radiation, and the purple blue fluorescent radiation and the original yellow fluorescent radiation are complementary to each other to form white light, so that the whiteness of the product under sunlight is improved. The fluorescent agent of the application can be naphthalene, anthracene, phenanthrene and other derivatives with relatively simple structures and fluorescent signals, or heterocyclic compounds with slightly large molecular weight and complex structures.

Surfactant (surfactant) means a substance added in a small amount to cause a significant change in the interfacial state of the solution system. Has immobilized hydrophilic and lipophilic groups, and can be oriented on the surface of the solution. The molecular structure of the surfactant has amphipathy: one end is hydrophilic group, and the other end is hydrophobic group; the hydrophilic group is usually a polar group such as carboxylic acid, sulfonic acid, sulfuric acid, amino or amine group and salts thereof, and hydroxyl group, amide group, ether bond and the like can also be used as the polar hydrophilic group; while hydrophobic groups are often nonpolar hydrocarbon chains, such as hydrocarbon chains of more than 8 carbon atoms. Surfactants are classified into ionic surfactants (including cationic surfactants and anionic surfactants), nonionic surfactants, amphoteric surfactants, built surfactants, and the like. The surfactant of the present application is selected from cationic surfactants, anionic surfactants, or nonionic surfactants. Preferably, the surfactant is selected from quaternary ammonium salts, carboxylates, alkyl sulfonates, alkylbenzene sulfonates or polysorbates. More preferably, the surfactant is selected from cetyltrimethylammonium bromide (CTAB), cetyltrimethylammonium chloride (CTAC), dodecyltrimethylammonium bromide (DTAB), dodecyltrimethylammonium chloride (DTAC), sodium Dodecyl Sulfate (SDS), sodium Tetradecyl Sulfate (STS), sodium hexadecyl sulfate, sodium octadecyl sulfate, potassium stearate, potassium oleate Sodium Dodecyl Benzene Sulfonate (SDBS), or tween 80.

According to another aspect of the present application, there is also provided a method for preparing a bioaerosol simulator, comprising the steps of:

1) Dissolving a fluorescent agent: adding the fluorescent agent into an organic solvent, and stirring and dissolving to obtain an organic solution of the fluorescent agent;

2) Dissolving a surfactant: adding a surfactant into water for dissolution to obtain an aqueous solution of the surfactant;

3) Mixing: mixing the solutions obtained in 1) and 2), and stirring;

4) Curing: and 3) standing and curing the solution obtained in the step 3) to obtain the bioaerosol simulator.

Wherein, in the step 1), the organic solvent is selected from polar solvent or nonpolar solvent.

Wherein the organic solvent is selected from methanol, ethanol, benzene or tetrahydrofuran.

Wherein the organic solvent in step 1) is a nonpolar solvent, and after step 3), the organic solvent is required to be left to stand and then separated to remove the organic solution which is not miscible with water.

Wherein, in the step 2), the addition amount of the surfactant is 1.05 to 1.20 times of the critical micelle concentration.

Wherein, in the step 3), the stirring time is 3-10min.

Wherein, in the step 4), standing and curing time is 6-72h.

As the number of surfactant molecules in solution increases, their hydrophobic portions attract each other, associate together, and the hydrophilic portions move toward water, with tens or more molecules associating together to form associated particles, known as micelles. Critical micelle concentration (critical micell concentration, CMC), the lowest concentration at which the surface active molecules associate to form micelles. The size of CMC is related to the structure, composition of the material. The surfactant is firstly easy to form micelle, and the addition amount of the surfactant is 1.05 to 1.20 times of the critical micelle concentration in consideration of cost.

Preferably, in step 3), the stirring time is 5-8min.

Preferably, in step 4), the standing and curing time is 12-48 hours.

Compared with the prior art, the application has the beneficial effects that:

1. the preparation method is simple, and the preparation of the simulator can be completed through the steps of dissolution, mixing, curing and the like; because common fluorescent agent is adopted, the solvent is easy to obtain, the cost is low, and the cost is greatly reduced;

2. the prepared simulator has strong fluorescent signal and good simulation test effect;

3. the concentration of the simulator is flexible and controllable, the simulators with various proportions can be prepared according to actual demands, and the obtained simulators can be diluted in any proportion to obtain the bioaerosol simulators with different concentrations;

4. the shape and the particle size of the biological simulator are controllable, different fluorescent compounds can be used for preparing biological aerosol simulators with different shapes and sizes, and the particle size of the aerosol can be controlled by adjusting the concentration of the simulators and the airflow of an aerosol generator. The fluorescent agent can be uniformly dispersed in the solution, and the particle size distribution of the fluorescent agent in the solution is little affected by the concentration, as shown in fig. 2. But the concentration has a significant effect on the particle size of the aerosol sprayed by the aerosol generator, and different solutions can simulate bioaerosols of different shapes and colors, see fig. 3.

5. The stability of the simulator is good, and FIG. 4 shows that the transparency and fluorescence intensity of the sample just prepared and stored for one month in dark place (the sample is irradiated by an ultraviolet lamp with the same intensity) are basically unchanged when the simulation agent is 0.1mmol/L anthracene aerosol.

Drawings

The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this specification, illustrate embodiments of the application and together with the description serve to explain the application. In the drawings:

FIG. 1 is a flow chart of the preparation of a bioaerosol mimetic agent of the present application;

FIG. 2 is a graph showing particle size distribution of a simulator in the state of solutions of different concentrations;

wherein FIG. 2 (a) is the particle size distribution in an anthracene-containing bioaerosol simulator solution at a concentration of 0.01 mmol/L;

FIG. 2 (b) is the particle size distribution in an anthracene-containing bioaerosol simulator solution at a concentration of 0.1 mmol/L;

FIG. 3 is an electron micrograph of the particle size distribution of an aerosol produced from solutions of different concentrations;

wherein FIG. 3 (a) is an electron micrograph of the particle size distribution of an anthracene-containing bioaerosol simulator at a concentration of 0.01 mmol/L;

FIG. 3 (b) is an electron micrograph of the particle size distribution of an anthracene-containing bioaerosol simulator at a concentration of 0.1 mmol/L;

FIG. 4 is a comparison of transparency and fluorescence intensity before and after one month of light-shielding preservation of the simulants;

wherein, FIG. 4 (a) is a transparency comparison of the simulant before and after one month of light-proof preservation;

FIG. 4 (b) is a comparison of fluorescence intensity before and after one month of light-shielding preservation of the mimetic agent;

FIG. 5 is a graph of the alarm response of 0.01mmol/L and 0.1mmol/L of anthracene-containing bioaerosol simulator.

Detailed Description

For the purpose of making the objects, technical solutions and advantages of the embodiments of the present application more apparent, the technical solutions in the embodiments of the present application will be clearly and completely described below, and it is apparent that the described embodiments are some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application. It should be noted that, without conflict, the embodiments of the present application and features of the embodiments may be arbitrarily combined with each other.

The bioaerosol simulants of the present application include fluorescent agents and surfactants. The fluorescent agent is selected from one or more of naphthalene, anthracene, phenanthrene and derivatives thereof, and heterocyclic compounds with fluorescent signals. The surfactant is selected from cationic surfactant, anionic surfactant or nonionic surfactant. Preferably, the surfactant is selected from quaternary ammonium salts, carboxylates, alkyl sulfonates, alkylbenzene sulfonates or polysorbates. More preferably, the surfactant is selected from cetyltrimethylammonium bromide (CTAB), cetyltrimethylammonium chloride (CTAC), dodecyltrimethylammonium bromide (DTAB), dodecyltrimethylammonium chloride (DTAC), sodium Dodecyl Sulfate (SDS), sodium Tetradecyl Sulfate (STS), sodium hexadecyl sulfate, sodium octadecyl sulfate, potassium stearate, potassium oleate Sodium Dodecyl Benzene Sulfonate (SDBS), or tween 80. As shown in fig. 1, the preparation method of the bioaerosol simulator comprises the following steps:

3) Mixing: mixing the solutions obtained in 1) and 2), and stirring;

In step 1), the organic solvent is selected from polar solvents or nonpolar solvents.

In step 1), the organic solvent is selected from methanol, ethanol, benzene or tetrahydrofuran.

The organic solvent in step 1) is a nonpolar solvent, and after step 3), it is necessary to stand and then separate the liquid to remove the organic solution which is not miscible with water.

In the step 2), the addition amount of the surfactant is 1.05 to 1.20 times of the critical micelle concentration.

In the step 3), the stirring time is 3-10min.

In the step 4), standing and curing time is 6-72h.

The bioaerosol simulator provided by the present application will be explained in detail by way of specific examples.

Example 1: preparation of anthracene-containing aerosol simulant

1) 35.6mg of anthracene is weighed by an electronic balance and added into 200mL of ethanol, and the anthracene is stirred and dissolved to obtain anthracene ethanol solution with the concentration of 1 mmol/L;

2) 18mg of cetyltrimethylammonium bromide (CTAB) is weighed by an electronic balance and added into 500mL of deionized water, and the mixture is stirred and dissolved to obtain CTAB aqueous solution;

3) Weighing 10mL of the solution prepared in the step 1), dropwise adding the solution into 100mL of the aqueous solution prepared in the step 2), vigorously stirring for 5min, standing and curing for 12h to obtain the anthracene-containing biological aerosol simulator with the concentration of about 0.1 mmol/L; in the same way, 1mL of the solution prepared in the step 1) is measured, and then added dropwise into 100mL of the aqueous solution prepared in the step 2), and the solution is vigorously stirred for 5min, and then left to stand and cure for 12h, so that the anthracene-containing bioaerosol simulator with the concentration of about 0.01mmol/L is obtained.

Example 2: preparation of aerosol simulants containing 4- (dimercaptomethylene) -2-methyl-6- (p-dimethylaminostyryl) -4H-pyran (DCM)

1) 60.6mgDCM is weighed by an electronic balance and added into 200mL tetrahydrofuran, and the mixture is stirred and dissolved to obtain DCM tetrahydrofuran solution with the concentration of 1 mmol/L;

2) 550mg of Sodium Dodecyl Sulfate (SDS) is weighed by an electronic balance and added into 200mL of deionized water, and the mixture is stirred and dissolved to obtain an aqueous solution of SDS;

3) Taking 100mL of the aqueous solution prepared in the step 2), heating to 50 ℃, taking 10mL of the solution prepared in the step 1), dropwise adding the solution into the aqueous solution at 50 ℃, vigorously stirring for 5min, standing and curing for 48h, and obtaining the biological aerosol simulator containing DCM with the concentration of about 0.1 mmol/L;

example 3: preparation of aerosol simulators containing 2,4, 5-Triphenylimidazole (TPI)

1) 59.2mgTPI is weighed by an electronic balance and added into 200mL of methanol, and the mixture is stirred and dissolved to obtain a TPI methanol solution with the concentration of 1 mmol/L;

2) 540mg of Sodium Dodecyl Benzene Sulfonate (SDBS) is weighed by an electronic balance and added into 200mL of deionized water, and stirred for dissolution, so as to obtain an aqueous solution of the SDBS;

3) 10mL of the solution prepared in the step 1) is measured and added dropwise into 100mL of the aqueous solution prepared in the step 2), and the solution is vigorously stirred for 5min, and then is stood for curing for 48h, so that the biological aerosol simulator containing TPI with the concentration of about 0.1mmol/L is obtained.

Example 4: preparation of rubrene-containing aerosol simulants

1) 53.3mg of rubrene is weighed by an electronic balance and added into 100mL of benzene, and the mixture is stirred and dissolved to obtain rubrene benzene solution with the concentration of 1 mmol/L;

2) Weighing 10mg of Tween 80 by an electronic balance, adding into 500mL of deionized water, stirring and dissolving to obtain an aqueous solution of Tween 80;

3) Weighing 10mL of the solution prepared in the step 1), dropwise adding the solution into 100mL of the aqueous solution prepared in the step 2), vigorously stirring for 5min, standing for 30min for layering, separating the solution, discarding the upper organic phase, standing and curing the lower aqueous phase for 12h, and obtaining the rubrene-containing biological aerosol simulator with the concentration of about 0.1 mmol/L.

Example 5: alarm response of anthracene-containing bioaerosol simulants at different concentrations

Taking anthracene-containing bioaerosol simulants with the concentration of 0.01mmol/L and 0.1mmol/L respectively, taking FCBR-100 independently researched and developed by a company as test equipment, sampling each sample for 3 seconds under the same test condition, and recording the indication change of fluorescent particles of a FCBR-100 bioaerosol detection module, wherein the test result is shown in figure 5. The device can sensitively detect the simulant with low concentration, and can respond obviously to simulants with different concentrations, and the recovery is rapid; the concentration of the simulant is increased, so that the response amplitude of fluorescent particles of the equipment can be remarkably improved.

Example 6: comparison of the simulation test results of the same concentration of the simulation agent prepared by the method of the application and the riboflavin

Taking anthracene-containing biological aerosol simulant (example 1) and riboflavin aqueous solution simulant with the concentration of 0.01mmol/L, carrying out generation by using the same biological aerosol generator, and testing the alarm response time of the two simulants to equipment and the maximum concentration value reached by the equipment after alarm under the same test condition by adopting an FCBR-100 biological aerosol detection module. Each sample was tested 3 times to obtain the test results shown in the following table. The simulant prepared by the method has quicker alarm response than the riboflavin, and the alarm response concentration of the equipment is far higher than that of the riboflavin.

As can be seen from the above examples and comparative examples, the present application provides a bioaerosol simulator with a flexible shape control, low cost and good fluorescence effect by mixing a fluorescent agent with a surfactant. The equipment can sensitively detect the low-concentration simulant prepared by the method, and the equipment can respond obviously and recover quickly for the simulant with different concentrations; the concentration of the simulant is increased, so that the response amplitude of fluorescent particles of the equipment can be remarkably improved. The simulant prepared by the method has quicker alarm response than the riboflavin, and the alarm response concentration of the equipment is far higher than that of the riboflavin.

The bioaerosol simulator in the application has the following advantages: 1. the preparation method is simple, and the preparation of the simulator can be completed through the steps of dissolution, mixing, curing and the like; because common fluorescent agent is adopted, the solvent is easy to obtain, the cost is low, and the cost is greatly reduced; 2. the prepared simulator has strong fluorescent signal and good simulation test effect, the equipment can sensitively detect the low-concentration simulator prepared by the method, and the equipment has high response speed and rapid recovery; 3. the concentration of the simulator is flexible and controllable, the simulators with various proportions can be prepared according to actual demands, and the obtained simulators can be diluted in any proportion to obtain the bioaerosol simulators with different concentrations; 4. the shape and the particle size of the biological simulator are controllable, the control method is simple, the biological aerosol simulators with different shapes and sizes can be prepared by using different fluorescent compounds, and the particle size of the aerosol can be controlled by adjusting the concentration of the simulators and the airflow of the aerosol generator. 5. The simulator has good stability and can be stored for more than one month in dark place.

Finally, it should be noted that: in this document, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.

The above embodiments are only for illustrating the technical solution of the present application, and are not limiting. Although the application has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A method for preparing a bioaerosol simulator, which is characterized in that the bioaerosol simulator comprises a fluorescent agent and a surfactant;

the fluorescent agent is selected from one of anthracene, 4- (dimercaptomethylene) -2-methyl-6- (p-dimethylaminostyryl) -4H-pyran or 2,4, 5-triphenylimidazole;

the surfactant is selected from cetyltrimethylammonium bromide, cetyltrimethylammonium chloride, dodecyltrimethylammonium chloride, sodium tetradecyl sulfate, sodium hexadecyl sulfate, sodium octadecyl sulfate or tween 80;

the preparation method of the bioaerosol simulator comprises the following steps:

2) Dissolving a surfactant: adding a surfactant into water for dissolution to obtain an aqueous solution of the surfactant, wherein the addition amount of the surfactant is 1.05-1.20 times of the critical micelle concentration;

3) Mixing: mixing the solutions obtained in 1) and 2), and stirring;

4) Curing: standing and curing the solution obtained in the step 3) for 12-48 hours to obtain the bioaerosol simulator;

the organic solvent in step 1) is a nonpolar solvent, and after step 3), the organic solvent is allowed to stand and then separated to remove the organic solution which is not miscible with water.