A heated cigarette smoke capture device and a heated cigarette sample pretreatment method

Abstract

The present invention belongs to the technical field of heated smoking utensils, and particularly relates to a heated cigarette smoke capture device and a heated cigarette sample pretreatment method. The smoke capture device comprises: a smoking machine, which has a Cambridge filter collector inside; a capture bottle, which is connected behind the Cambridge filter collector of the smoking machine, the lower part of the capture bottle is filled with a capture liquid, the upper part of the capture bottle has a filter paper circumferentially attached to the inner wall of the capture bottle, and the filter paper is wetted by the capture liquid as a whole. The present invention also discloses a heated cigarette sample pretreatment method. The present invention designs a smoke capture bottle that simulates the human oral cavity for the first time, and the filter paper wetted by the capture liquid is attached to the inner wall of the upper part of the capture bottle. The capture bottle is set in a water bath device at 36.7-37.7 degrees Celsius, so that the rough and moist surface of the human oral cavity and the process of smoke absorption in the saliva of the oral cavity can be more realistically simulated.

Description

Technical Field

[0001] The invention belongs to the technical field of heated smoking articles, and in particular relates to a heated cigarette smoke capture device and a heated cigarette sample pretreatment method. Background Art

[0002] The increasingly stringent tobacco regulatory legislation and the in-depth research on smoking and health have led tobacco companies to seek new tobacco products with lower risks and no environmental smoke. Heated cigarettes are characterized by heating tobacco instead of burning it, providing consumers with a certain tobacco characteristic experience. Heated cigarette products avoid the harm caused by the complex mixture produced when traditional cigarette products are burned. At this stage, it has become one of the new directions for foreign tobacco companies to turn away from traditional cigarette products.

[0003] At present, the samples of heated cigarettes used for biological effect testing are mainly exposed directly to whole smoke and indirectly exposed after capturing total particulate matter (TPM, hereinafter referred to as particulate matter) in smoke. Both methods have their own advantages and disadvantages. The whole smoke direct exposure method can more comprehensively reflect the potential effects of the sample, but because the whole smoke direct exposure has high requirements for instruments, it is not conducive to promotion. Although the indirect exposure after capturing TPM has a capture efficiency of more than 99%, some gas phase substances (GVP) cannot be captured on the Cambridge filter.

[0004] In the process of realizing the present invention, the inventors found that although there is a method for capturing gas phase substances, which is to connect an ice bath glass trap containing phosphate buffer solution after the Cambridge filter trap. However, when performing cytotoxicity tests, for convenience, the TPM captured by the Cambridge filter and the GVP captured by the ice bath glass trap are separately subjected to cytotoxicity detection experiments.

[0005] The inventors found that the above method has the following problems:

[0006] 1. The trapping solution of the ice bath glass trap is generally a phosphate buffer solution. When the trapping solution is added to the cell culture medium in the cytotoxicity test, the chemical ions in the trapping solution will affect the normal growth of the cells and interfere with the cytotoxicity test results.

[0007] 2. Since TPM and GVP were tested separately for cytotoxicity, it is impossible to obtain the overall effect of TPM and GVP in the smoke of a single cigarette on cell growth and the toxicity test results. Therefore, it is impossible to objectively and comprehensively evaluate the potential effects of heated cigarette smoke.

[0008] The present invention is proposed to solve the above-mentioned problems. Summary of the invention

[0009] In view of the shortcomings of the prior art, the present invention provides a heated cigarette smoke capture device and a heated cigarette sample pretreatment method. Cambridge filter capture is combined with capture bottle capture to simultaneously obtain particulate matter and gaseous matter for back-end biological effect testing, thereby objectively and comprehensively evaluating the potential effects of heated cigarette smoke.

[0010] A first aspect of the present invention provides a heated cigarette smoke capture device for biological effect testing, the smoke capture device comprising:

[0011] a smoking machine with a Cambridge filter trap inside;

[0012] The collecting bottle is connected behind the Cambridge filter collector of the smoking machine. The lower part of the collecting bottle is filled with collecting liquid. The upper part of the collecting bottle has filter paper circumferentially attached to the inner wall of the collecting bottle. The filter paper is wetted by the collecting liquid as a whole.

[0013] The purpose of wetting the filter paper here is to imitate the rough and moist surface structure of the human oral cavity, so that the process of simulating the rough and moist surface of the human oral cavity and the absorption of smoke in the saliva of the oral cavity is closer to the real situation.

[0014] Preferably, the lower end of the filter paper does not contact the capture liquid, and the filter paper completely covers the portion of the inner wall of the capture bottle body that does not contact the capture liquid.

[0015] Preferably, the capture bottle is placed in a water bath at 36.7-37.7 degrees Celsius. The purpose here is to simulate the temperature environment of the human oral cavity.

[0016] Previously, the condition for using a capture bottle to capture the gas phase of flue gas was that the capture bottle was placed in an ice bath device in order to condense the gas phase.

[0017] Preferably, the capture liquid is a cell culture medium. Of course, the cell culture medium is a liquid cell culture medium.

[0018] The second aspect of the present invention provides a method for pre-treating heated cigarette samples for biological effect testing, wherein the method uses the smoke capture device of the first aspect, and the method comprises the following steps:

[0019] A. smoking a heated cigarette using the smoking machine of the first aspect, the heated cigarette generates a smoke sample, the smoke sample passes through the Cambridge filter collector and the capture bottle in sequence, the particulate matter in the smoke sample is captured by the Cambridge filter in the Cambridge filter collector, and the gaseous matter in the smoke sample is captured by the capture liquid and filter paper in the capture bottle;

[0020] B. soaking the Cambridge filter disc that has captured the particulate matter in dimethyl sulfoxide (DMSO) for extraction to obtain a dimethyl sulfoxide solution containing the particulate matter as a particulate matter storage solution;

[0021] The filter paper that has captured the gas phase substance is soaked in the capture liquid in the capture bottle, and extracted to obtain the capture liquid containing the gas phase substance;

[0022] C. According to the cytotoxicity preliminary experiment, the particulate matter storage solution described in step B is diluted to the concentration of the particulate matter working solution, and the number of cigarettes smoked during the capture process of the Cambridge filter collector and the capture bottle is used as a measurement scale. According to the concentration of the particulate matter working solution, the capture solution containing the gas phase substance is diluted with the capture solution in a certain proportion to obtain a diluted gas phase substance capture solution;

[0023] D. Mix the particulate matter working solution of step C and the diluted gas phase material capture solution for the back-end biological effect test.

[0024] Preferably, in step A, the smoking machine according to claim 1 is used to smoke heated cigarettes, and multiple smoking channels containing the Cambridge filter collector are used to smoke multiple cigarettes simultaneously, the Cambridge filter collector of at least one smoking channel of the smoking machine is connected to the capture bottle, and the number of cigarettes captured by the Cambridge filter collector and the number of cigarettes captured by the capture bottle are the same or different.

[0025] Preferably, the capture solution is a cell culture medium corresponding to the test cells used in the back-end biological effect test.

[0026] Preferably, in step C, the method for calculating the dilution ratio of the captured liquid containing the gas phase substance according to the concentration of the particulate phase substance working liquid is as follows:

[0027] The total number of cigarettes smoked during the Cambridge filter trapping process is recorded as b;

[0028] The total mass of the particulate matter captured by the Cambridge filter disc during the Cambridge filter disc capture process is recorded as aug;

[0029] The concentration of the particle phase material working solution is recorded as dug particle phase material / ml dimethyl sulfoxide;

[0030] The concentration of the gas phase substance in the capture liquid in step B is recorded as e cigarettes / ml capture liquid;

[0031] Then in step C, the dilution factor of the capture liquid containing the gas phase substance is:

[0032] In this article, the GVP concentration in the capture liquid is calculated as: number of smoked cigarettes / capture liquid volume, so the unit is: cigarettes / ml.

[0033] The present invention has the following beneficial effects:

[0034] 1. The present invention is the first to design a smoke collection bottle that simulates the human oral cavity. A filter paper moistened with a collection liquid is attached to the inner wall of the upper part of the collection bottle. The collection bottle is placed in a water bath device at 36.7-37.7 degrees Celsius, which can more realistically simulate the process of smoke absorption by the rough and wet surface of the human oral cavity and the saliva of the oral cavity. The prior art directly uses an ice bath collector to capture smoke gas phase substances, and has never thought of improving it to be closer to the process of smoke absorption by the human oral cavity.

[0035] 2. The present invention uses the cell culture medium corresponding to the cells used for biological effect testing as the capture liquid in the capture bottle for the first time. This method avoids the influence of ions in the general phosphate buffer capture liquid on cell growth and will not interfere with the biological effect test results at the back end.

[0036] 3. The present invention is the first to mix two sample liquids containing TPM and GVP in a certain ratio, and when used in back-end testing, it can objectively and effectively reflect the comprehensive biological effects of TPM and GVP.

[0037] In addition, the present invention considers diluting the GVP according to the number of smoked cigarettes for the first time, and proposes a method for calculating the dilution ratio for the first time, taking the number of cigarettes smoked in the Cambridge filter collector and the capture bottle capture process as a measure, and diluting the capture liquid containing the gas phase substance with the capture liquid according to the concentration of the particulate matter working liquid in a certain ratio to obtain the diluted gas phase substance capture liquid for the back-end biological effect test. This method avoids the defect that the gas phase substance is infinitely enriched during capture, resulting in too high pH value of the capture liquid, pH value affects the survival rate of cells, and the harmfulness of the substance itself cannot be detected.

[0038] 4. The present invention comprehensively examines the exposure pathways and modes of action of heated cigarettes for the first time, establishes a relatively complete method for pre-treatment of heated cigarette smoke, and effectively treats the samples, so that the samples obtained by the method of the present invention can objectively and effectively reflect the true state of the biological effects of heated cigarette smoke when used in the back-end biological effect test. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is the gas phase material (GVP) collection bottle of the sample in Example 1.

[0040] Figure 2Graph showing the cytotoxicity test results of TPM, GVP, and TPM+GVP of heated cigarette samples and reference cigarette samples under different smoking modes in Example 1.

[0041] Figure 3 These are the cytotoxicity test results of the gas phase substances (GVP) of the four samples of Comparative Example 1.

[0042] List of reference numerals:

[0043] 1. Inlet pipe, 11. Outlet head, 2. Inlet and outlet pipe covers, 3. Outlet pipe, 31. Expansion section, 4. Filter paper. DETAILED DESCRIPTION

[0044] The content of the present invention is further explained below through specific implementation examples.

[0045] Example 1

[0046] A heated cigarette smoke capture device for biological effect testing, the smoke capture device comprising:

[0047] a smoking machine with a Cambridge filter trap inside;

[0048] The collecting bottle is connected behind the Cambridge filter collector of the smoking machine. The lower part of the collecting bottle is filled with collecting liquid. The upper part of the collecting bottle has filter paper 4 circumferentially attached to the inner wall of the collecting bottle. The filter paper 4 is wetted by the collecting liquid.

[0049] The lower end of the filter paper 4 does not contact the capture liquid, and the filter paper 4 completely covers the portion of the inner wall of the capture bottle body that does not contact the capture liquid.

[0050] The capture bottle is placed in a water bath at 37 degrees Celsius. The capture liquid is a liquid cell culture medium.

[0051] The specific design of the capture bottle is as follows Figure 1 As shown, it includes an upper air inlet and outlet pipe cover 2 and a lower collection bottle body. The filter paper 4 is attached to the inner wall of the collection bottle body in a circumferential direction. The collection liquid at the lower end of the filter paper 4 is not Figure 1 Indicated in.

[0052] The upper air inlet and outlet pipe cover 2 comprises:

[0053] The spherical expansion part is mushroom-shaped and has a mushroom head and a mushroom stem, wherein the mushroom head at the upper end is sealed and the mushroom stem at the lower end is open and plugged into the collection bottle body;

[0054] An air inlet pipe 1, the air inlet pipe 1 passes through the spherical expansion part, the upper end of the air inlet pipe 1 extends out of the upper air inlet and outlet pipe cover 2 for connecting with the external air inlet channel, and the lower end goes deep into the interior of the collection bottle body, the lower end of the air inlet pipe 1 has an air outlet head 11 connected to the pipe body in one piece, the air outlet head 11 is a hollow sphere, and the surface of the sphere has a plurality of air outlet holes;

[0055] The air outlet pipe 3 is connected to the mushroom head of the spherical expansion part and extends out of the upper air inlet and outlet pipe cover 2. The air outlet pipe 3 is provided with an expansion section 31.

[0056] One function of the expanded section 31 on the air outlet pipe 3 is to facilitate differentiation from the air inlet pipe 1 during installation.

[0057] The gas outlet head 11 is immersed in the collecting liquid. The gas outlet head 11 is designed to make the ejected gas more dispersed, which is conducive to the collecting liquid in the collecting bottle to absorb the gas more fully.

[0058] The process of capturing GVP in flue gas samples by the capture bottle is as follows:

[0059] When in use, the external air intake channel is connected to the upper end of the air intake pipe 1, and the smoke sample enters the air intake pipe 1 from the air intake channel, and is sprayed into the collecting liquid in the collecting bottle through a plurality of air outlet holes on the spherical surface of the air outlet head 11, and part of the smoke sample is absorbed by the collecting liquid, and the rest of the smoke sample diffuses into the cavity part at the upper part of the collecting bottle, and part of the smoke sample is again absorbed by the filter paper 4 moistened by the inner wall of the collecting bottle, and the rest of the smoke sample diffuses to the outside of the collecting bottle through the spherical expansion part and the air outlet pipe 3.

[0060] Example 2

[0061] A method for pre-treating heated cigarette samples for biological effect testing comprises the following steps:

[0062] TPM and TPM+GVP samples of heated cigarette samples and reference cigarettes were prepared. The heated cigarette sample was 1# and the reference cigarette sample was 2#. The preparation methods and smoking parameters of the two samples to be tested are shown in Table 1.

[0063] Table 1. Preparation methods and suction parameters of two samples to be tested

[0064]

[0065] The specific sample pretreatment method is as follows:

[0066] 1. Determine the number of puffs based on the heating time and puffing pattern of the heated cigarette.

[0067] In this embodiment, the number of puffs of the two samples to be tested under different puffing modes is shown in Table 2, the number of puffs of the heated cigarette is the set value, and the number of puffs of the reference cigarette is the actual puff value.

[0068] Table 2 Number of puffs of two samples under different puff modes

[0069]

[0070] 2. Use a smoking machine to smoke and collect the TPM and corresponding GVP of each sample at the same time. The GVP is captured by a capture bottle (capture bottle such as Figure 1 The capture liquid was culture medium, and TPM was captured using Cambridge filter discs.

[0071] 3. Capture according to the key sample preparation parameters in Table 3.

[0072] Table 3 Key parameters for cigarette sample preparation

[0073]

[0074]

[0075] 4. Record the number of puffs during the puffing process.

[0076] The Cambridge filter disc that has captured the particulate matter is soaked in dimethyl sulfoxide (DMSO) for extraction to obtain a dimethyl sulfoxide solution containing the particulate matter as a particulate matter storage solution;

[0077] The filter paper that has captured the gas phase substance is soaked in the capturing liquid in the capturing bottle, and extracted to obtain a capturing liquid containing the gas phase substance.

[0078] (1)Sample 1#-I:

[0079] Cambridge filter trap collection process:

[0080] Smoked 18 heated cigarettes, taking 5 puffs per cigarette;

[0081] Six Cambridge filters were used, and the total mass of TPM captured by the Cambridge filter was 0.47 g. The volume of DMSO used for extraction was 23.5 ml. The TPM concentration in DMSO after extraction was 20 mg / ml, which was used as the storage solution. When it was used, according to the cytotoxicity pre-experiment and the toxicity of TPM in the cigarette sample, the TPM concentration was diluted to an appropriate dose concentration of 600 ug / ml as the working solution for the cytotoxicity test.

[0082] Capture bottle capture process:

[0083] 12 heated cigarettes were smoked, with 5 puffs per cigarette, the volume of the captured liquid was 20 ml, and the GVP concentration was 0.6 cigarettes / ml.

[0084] During the smoking test of each sample, the Cambridge filter was replaced according to the number of cigarettes smoked, and only one capture bottle was used and was not replaced.

[0085] (2) Sample 1#-H:

[0086] Cambridge filter trap collection process:

[0087] Smoked 18 heated cigarettes, taking 10 puffs per cigarette;

[0088] Six Cambridge filters were used, and the total mass of TPM captured by the Cambridge filter was 0.5212 g. The volume of DMSO used for extraction was 26.1 ml. The TPM concentration in DMSO after extraction was 20 mg / ml, which was used as the storage solution. When it was used, according to the cytotoxicity pre-experiment and the toxicity of TPM in cigarette samples, the TPM concentration was diluted to an appropriate dose concentration of 600 ug / ml, which was used as the working solution for cytotoxicity testing.

[0089] Capture bottle capture process:

[0090] 12 heated cigarettes were smoked, with 10 puffs per cigarette, the volume of the captured liquid was 20 ml, and the GVP concentration was 0.6 cigarettes / ml.

[0091] (3)Sample 2#-I:

[0092] Cambridge filter trap collection process:

[0093] Smoked 30 heated cigarettes, taking 8.5 puffs per cigarette;

[0094] Six Cambridge filters were used, and the total mass of TPM captured by the Cambridge filter was 0.2485 g. The volume of DMSO used for extraction was 24.9 ml. The TPM concentration in DMSO after extraction was 10 mg / ml, which was used as the storage solution. When it was used, according to the cytotoxicity pre-experiment and the toxicity of TPM in the cigarette sample, the TPM concentration was diluted to an appropriate dose concentration of 200 ug / ml as the working solution for the cytotoxicity test.

[0095] Capture bottle capture process:

[0096] Five heated cigarettes were smoked, with the number of puffs being 8.5 per cigarette, the volume of the capture liquid being 50 ml, and the GVP concentration being 0.1 cigarette per ml.

[0097] (4) Sample 2#-H:

[0098] Cambridge filter trap collection process:

[0099] 18 heated cigarettes were smoked, with 13.2 puffs per cigarette.

[0100] Six Cambridge filters were used, and the total mass of TPM captured by the Cambridge filter was 0.4328 g. The volume of DMSO used for extraction was 43.3 ml, and the TPM concentration in DMSO after extraction was 10 mg / ml; it was used as a storage solution. When it was used, according to the cytotoxicity pre-experiment and the toxicity of TPM in cigarette samples, the TPM concentration was diluted to an appropriate dose concentration of 200 ug / ml, and used as a working solution for cytotoxicity testing.

[0101] Capture bottle capture process:

[0102] Three heated cigarettes were smoked, with the number of puffs being 8.5 per cigarette, the volume of the capture liquid being 30 ml, and the GVP concentration being 0.1 cigarette per ml.

[0103] 5. Using the number of cigarettes smoked during the Cambridge filter trap and the trap bottle capture process as a measure, and according to the concentration of the particulate matter working solution, dilute the four capture solution samples containing the gas phase substance with liquid cell culture medium in a certain proportion to obtain a diluted gas phase substance capture solution;

[0104] The particulate matter working solution and the diluted gas phase matter capture solution are mixed to obtain a TPM+GVP sample for a back-end biological effect test.

[0105] TPM is extracted with DMSO and its unit is ug particulate matter / ml dimethyl sulfoxide, while GVP is captured by liquid cell culture medium and its unit is cigarette / ml capture liquid.

[0106] The calculation method of GVP dilution multiple is as follows: Taking sample 1#-I as an example

[0107] According to conventional methods in the art, after a cytotoxicity pre-experiment, the TPM sample working solution concentration of sample 1#-1 was 600ug / ml, and the detection concentrations were 0ug / ml, 60ug / ml, 120ug / ml, 180ug / ml, 240ug / ml, 320ug / ml, 360ug / ml, 420ug / ml, 480ug / ml, 540ug / ml, and 600ug / ml. That is, within this range, the cell survival rate was well correlated with the concentration of TMP. The working solution concentration of each sample was related to the toxicity of the sample TPM.

[0108] The TPM concentration of sample 1#-I is: 0.47*1000*1000 / 18=26111.11ug / vial;

[0109] Calculated by the number of cigarettes, the TPM concentration is: 600 / 26111.11=0.0229787 cigarettes / ml;

[0110] Because: GVP sample concentration is 0.6 tube / ml,

[0111] Therefore: The dilution factor of the GVP sample is: 0.6 / 0.0229787=26.11

[0112] Therefore, the GVP sample was diluted 26 times and mixed with the TPM sample in a 1:1 ratio for testing.

[0113] Another way to calculate the dilution factor is to directly insert it into the formula of the present invention:

[0114]

[0115] According to the above calculation method, the GVP sample dilution factor of sample 1#-H is 29, the GVP sample dilution factor of sample 2#-I is 4.14, and the GVP sample dilution factor of sample 2#-H is 12.

[0116] Example 3

[0117] The TPM working solution sample prepared in step 4 of Example 2 and the TPM+GVP sample prepared in step 5 were subjected to cytotoxicity test.

[0118] The test method is: MTT cytotoxicity method. The test cells are RPMI2650 human nasal epithelial cells.

[0119] Figure 2 Effects of TPM and TPM+GVP samples of heated cigarette samples and reference cigarettes on cell viability

[0120] Figure 2 In the above, the TPM working solution sample prepared in step 4 is recorded as: T, and the TPM+GVP sample prepared in step 5 is recorded as: T+V. The abscissa represents the concentration of TPM in all samples, and the unit is: ug particulate matter / ml dimethyl sulfoxide.

[0121] The TPM working solution sample of sample 1# prepared in step 4 of Example 2 and the TPM+GVP sample prepared in step 5 were diluted again to 0-600ug particulate matter / ml dimethyl sulfoxide (specific detection concentrations: 0ug / ml, 60ug / ml, 120ug / ml, 180ug / ml, 240ug / ml, 320ug / ml, 360ug / ml, 420ug / ml, 480ug / ml, 540ug / ml, 600ug / ml) for testing.

[0122] The TPM working solution sample of sample 2# prepared in step 4 of Example 2 and the TPM+GVP sample prepared in step 5 were diluted again to 0-200ug particulate matter / ml dimethyl sulfoxide (detection concentrations: 0ug / ml, 20ug / ml, 40ug / ml, 60ug / ml, 80ug / ml, 100ug / ml, 120ug / ml, 1,40ug / ml, 160ug / ml, 180ug / ml, 200ug / ml.) for testing. The diluents were all the liquid cell culture medium of Example 2, which was sourced from commercially available RPMI1640 cell culture medium.

[0123] Depend on Figure 2 It can be seen that:

[0124] 1. As the sample test dose increases, the cell survival rate of the test cells shows a downward trend.

[0125] 2. There was no significant difference in the effects of TPM samples and TPM+GVP mixed samples on the survival rate of the test cells, which indicated that GVP had no significant effect on the survival rate of the test cells.

[0126] This is consistent with the understanding of technicians in this field: the gas phase substances in the smoke inhaled by normal people after being intercepted by the Cambridge filter should have little effect on the survival rate of cells.

[0127] Therefore, the calculation of the dilution multiple of GVP and the dilution multiple of TMP storage solution cannot be simply diluted in proportion, but needs to be determined through experimental exploration and calculation to determine the appropriate ratio. The mixture of GVP and TMP cannot be simply diluted in proportion, but needs to be determined through experimental exploration and calculation to determine the appropriate ratio.

[0128] Comparative Example 1

[0129] The gas phase substances of the four samples obtained in step 5 of Example 2 were diluted according to random proportions and directly used for cytotoxicity testing.

[0130] The test method is: MTT cytotoxicity method. The test cells are RPMI2650 human nasal epithelial cells.

[0131] Figure 3 The cytotoxicity test results of the gas phase substances (GVP) of the four samples of Comparative Example 1 are shown below. The horizontal axis is the dosage, and the unit is V capture solution / V diluent (liquid cell culture medium). The vertical axis is the cell survival rate, and the unit is %.

[0132] Figure 3Results: There is no specific pattern between cell survival rate and dilution multiple. As the dilution multiple decreases, the cell survival rates of two samples (samples 2#-I and 2#-H) drop rapidly to 0, the cell survival rate of one sample (sample 1#-H) decreases slowly, and the cell survival rate of another sample (sample 1#-I) even exceeds 100%.

[0133] The gas phase substances intercepted by Cambridge filter in the smoke inhaled by normal people should have little effect on the survival rate of cells. Therefore, the cell survival rate should be close to 100%.

[0134] This comparative example 1 results in Figure 3 The possible reason is that the gas phase material is infinitely enriched during the capture, resulting in a too high pH value of the capture solution. It is the pH value that affects the survival rate of the cells, not the harmfulness of the material itself. It can be seen that when the two samples of GVP and TPM are mixed, the appropriate dilution ratio of GVP is necessary to objectively reflect the true safety of the product.

[0135] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A method for pre-treatment of heated cigarette samples for biological effect testing, characterized in that: The sample pretreatment method uses the following fume capture device, which includes: a smoking machine with a Cambridge filter trap inside; A collection bottle connected after the Cambridge filter collector of the smoking machine, wherein the lower part of the collection bottle is filled with a collection liquid, and the upper part of the collection bottle has a filter paper circumferentially attached to the inner wall of the collection bottle, and the filter paper is entirely wetted by the collection liquid; The sample pretreatment method comprises the following steps: A. Using the smoking machine described above to smoke heated cigarettes, the heated cigarettes generate smoke samples, the smoke samples pass through the Cambridge filter collector and the capture bottle in sequence, the particulate matter in the smoke sample is captured by the Cambridge filter in the Cambridge filter collector, and the gaseous matter in the smoke sample is captured by the capture liquid and filter paper in the capture bottle; B. soaking the Cambridge filter disc that has captured the particulate matter in dimethyl sulfoxide for extraction to obtain a dimethyl sulfoxide solution containing the particulate matter as a particulate matter storage solution; The filter paper that has captured the gas phase substance is soaked in the capture liquid in the capture bottle, and extracted to obtain the capture liquid containing the gas phase substance; C. According to the cytotoxicity preliminary experiment, the particulate matter storage solution described in step B is diluted to the concentration of the particulate matter working solution, and the number of cigarettes smoked during the capture process of the Cambridge filter collector and the capture bottle is used as a measurement scale. According to the concentration of the particulate matter working solution, the capture solution containing the gas phase substance is diluted with the capture solution in a certain proportion to obtain a diluted gas phase substance capture solution; D. Mixing the particulate matter working solution of step C and the diluted gas phase material capture solution for a back-end biological effect test; In the step C, the method for calculating the dilution ratio of the capture liquid containing the gas phase substance according to the concentration of the particulate phase substance working liquid is as follows: The total number of cigarettes smoked during the Cambridge filter trapping process is recorded as b; The total mass of the particulate matter captured by the Cambridge filter disc during the Cambridge filter disc capture process is recorded as aug; In step C, the concentration of the particle phase material working solution is recorded as d ug particle phase material / ml dimethyl sulfoxide; The concentration of the gas phase substance in the capture liquid in step B is recorded as e cigarettes / ml capture liquid; Then in step C, the dilution factor of the capture liquid containing the gas phase substance is:

2. The smoke capture device according to claim 1, characterized in that: The lower end of the filter paper does not contact the capture liquid, and the filter paper completely covers the portion of the inner wall of the capture bottle body that does not contact the capture liquid.

3. The smoke capture device according to claim 1, characterized in that: The catch flask was placed in a water bath at 36.7-37.7 degrees Celsius.

4. The smoke capture device according to claim 1, characterized in that: The capture solution is a cell culture medium.

5. The sample pretreatment method according to claim 1, characterized in that: In the step A, the smoking machine according to claim 1 is used to smoke heated cigarettes, and multiple smoking channels containing the Cambridge filter collector are used to smoke multiple cigarettes at the same time, the Cambridge filter collector of at least one smoking channel of the smoking machine is connected to the capture bottle, and the number of cigarettes captured by the Cambridge filter collector and the number of cigarettes captured by the capture bottle are the same or different.

6. The sample pretreatment method according to claim 1, characterized in that: The capture solution is the cell culture medium corresponding to the test cells used in the back-end biological effect test.

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