Trapping method and cytotoxicity testing method for smoke aerosol of heating non-combustible cigarette

A heat-not-burn, cigarette smoke technology, applied in the preparation of test samples, the determination/inspection of microorganisms, biochemical equipment and methods, etc.

Pending Publication Date: 2020-11-03
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In some existing technologies for trapping smoke gaseous phase substances used in cigarette smoke chemical analysis experiments, organic acids are used as gaseous phase substance leaching solutions, and lye such as sodium hydroxide is also used to adjust the pH value. These organic acids Alkaline and lye itself will cause damage to the cells. In addition, organic acids will react with some chemical components in the smoke, so that the chemical composition of the smoke will change during the subsequent toxicology test, which cannot truly reflect the actual smoke. Biological effects of ingredients
In some existing technologies for capturing gaseous phase substances in cigarette smoke cell experiments, the steps are relatively cumbersome, for example, the gaseous phase substances are captured by phosphate buffer solution (PBS) first, and then organic solvents are used for capture. Multi-stage capture method, and ice bath conditions a

Method used

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  • Trapping method and cytotoxicity testing method for smoke aerosol of heating non-combustible cigarette
  • Trapping method and cytotoxicity testing method for smoke aerosol of heating non-combustible cigarette
  • Trapping method and cytotoxicity testing method for smoke aerosol of heating non-combustible cigarette

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Embodiment 1

[0031] Embodiment of the present embodiment is not heated cigarette smoke aerosol-combustion methods, which processes such as figure 1 As shown, including the following steps:

[0032] (1) linear smoking machine three heating the cigarette does not burn the test sample 1 using the suction mode of the suction depth of Canada (suction bell curve, the suction interval of 30s, each of capacity 55mL suction port, each port suction duration 2s), during suction diameter Cambridge filter capture 44mm total particulate matter smoke per puff by the time the gas was passed into Cambridge filter 15mL DMSO solution containing 4 5, the absorption bottle;

[0033] When the suction, each suction heat does not burn the cigarette 10, each of the Cambridge filter trap 4 total heating without burning cigarette smoke particle phase was prepared in a Cambridge filter 5 samples;

[0034] (2) after the trap, the five Cambridge filter was transferred to a conical flask 2 6, and the gas absorption bottle c...

Embodiment 2

[0040]In this example, the extracellular toxicity test is performed on the extravasation of the flue gas sol obtained in Example 1, and the trial cell is a 1549 cell line, and the CCK-8 kit test is used, and the specific test method is: 1) will be prepared. Cell suspension (15000 / well) in 56-well plates, cultured 24h; 2) Remove medium in the culture plate, adding a control medium and medium containing different concentrations of flue gas sol extract medium ( 100 μl / well), incubation under 37 ° C; 3) Remove the solution in the culture plate, add 100 μl of the configured CCK-8 solution to each well (10 μl of CCK-8 mother liquor per 100 μl), 37 ° C Incubation 0.5-2H; 4) Shake 5 min at room temperature, and the absorbing value was measured under 450 nm with an enzyme label. Cell survival rate = (experimental pore absorbance average / control pore absorbance average) × 100%. Test results figure 2 Indicated.

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Abstract

The invention belongs to the technical field of tobacco toxicity analysis, and particularly relates to a trapping method and a cytotoxicity testing method for smoke aerosol of a heat-not-burn cigarette. The method for capturing the aerosol in the smoke of the heat-not-burn cigarette comprises the following steps: (1) in the process of smoking the heat-not-burn cigarette, capturing total particulate matters in the smoke by adopting a Cambridge filter, and absorbing smoke gas phase matters penetrating through the Cambridge filter by adopting dimethyl sulfoxide; and after the capture is finished,extracting the smoke total particulate matters captured on the Cambridge filter by adopting dimethyl sulfoxide, and forming a smoke aerosol capture object by the dimethyl sulfoxide with the smoke total particulate matters extracted and the dimethyl sulfoxide with the smoke gas-phase matters absorbed in the step (1). The invention establishes a novel smoke gas solvent trapping method for in-vitrotoxicity test of the heat-not-burn cigarette, achieves comprehensive trapping of smoke aerosol of the heat-not-burn cigarette, and provides a basis for comprehensive evaluation of toxicity test of theheat-not-burn cigarette.

Description

Technical field [0001] The present invention belongs to the technical field of tobacco toxicity analysis, in particular to a method of heating without combustion of the cigarette smoke aerosol trap and cytotoxicity testing methods. Background technique [0002] Heating the cigarette does not burn with the "heating rather than burning tobacco or tobacco sheet" characteristics, as compared with conventional cigarettes, occur in different ways aerosol fumes, without cracking the combustion process, reducing the harmful components of tobacco combustion temperature cleavage of the chemical composition of mainstream smoke emission is greatly reduced; compared with the electronic cigarette, tobacco feature can provide some experience for consumers. Based on the lower heating without burning harmful components of cigarette tobacco release characteristics and has the advantage of feeling, which is considered to be a promising new type of cigarette. Currently, in vitro toxic effects of cig...

Claims

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Application Information

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IPC IPC(8): G01N1/40C12Q1/02
CPCG01N1/4055G01N33/5014G01N2001/4061
Inventor 李翔赵俊伟尚平平华辰凤赵阁崔华鹏谢复炜刘惠民
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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