Detection method for cytotoxicity of gas-liquid contact type cigarettes under full-smoke exposure
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A technology of cytotoxicity and gas-liquid contact, applied in measuring devices, air quality improvement, instruments, etc., can solve the problems of undiscovered public reports, etc., and achieve the effect of low cost
Active Publication Date: 2012-06-13
YUNNAN RES INST OF TOBACCO SCI
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Embodiment 1
[0033] Example 1: Detection of Neutral Red Blood Cell Toxicity under Total Smoke Exposure of a Mixed Cigarette
[0034] Prepare a CHO cell suspension according to the above steps a., and then follow the above steps b., c. and d. for total cigarette smoke exposure of CHO cells. When the cigarette sample is smoked, mainstream cigarette smoke is produced and then diluted with clean air and introduced into ZL201120063251 In the full smoke exposure bottle of .6, set 4 different clean air flow rates (0mL / min, 250mL / min, 500mL / min, 750mL / min, 1000mL / min) to dilute mainstream cigarette smoke. The exposure time was 5min, the cigarette smoking volume was 35mL, the smoking duration was 2s, and the smoking interval was 58s.
[0035] After the smoke exposure is completed, perform the neutral red blood cell toxicity test according to the above step e.(1), and calculate the cell inhibition rate CI according to the absorbance value.
[0036] CI = ( 1 - OD n - OD o O...
Embodiment 2
[0046] Example 2: Detection of MTT cytotoxicity of a certain mixed cigarette under total smoke exposure
[0047] Except that the MTT cytotoxicity test is performed according to step e.(2), other experimental methods are the same as in Example 1. The MTT cytotoxicity test results of a certain mixed cigarette under total smoke exposure are shown in Tables 3 and 4, and the dose response relationship between smoke exposure and cytotoxicity in the MTT cytotoxicity test under total smoke exposure is analyzed, see figure 2 .
[0048] From Table 3 and figure 2 It can be seen that as the flow rate of clean air increases, the cell inhibition rate produced by flue gas decreases, showing a significant dose-response relationship.
[0049] Table 3 MTT cytotoxicity test results of a certain mixed cigarette under total smoke exposure
[0050]
[0051] Table 4 Smoke dilution ratio and cell inhibition rate in MTT cytotoxicity test
[0052]
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Abstract
The invention relates to a detection method for the cytotoxicity of gas-liquid contact type cigarettes under full-smoke exposure, belonging to the technical field of safety evaluation of cigarette smoke. The detection method provided by the invention comprises the following steps of: firstly, setting cigarette suction parameters to carry out suction on a cigarette; diluting the smoke by clean air with the flow speed of 0-1000 mL/min and transferring the diluted cigarette full smoke into a full-smoke exposure bottle disclosed in the patent with the patent number of ZL201120063251.6 to come into contact and exposure with suspension cells in the bottle, wherein the exposure time is 5 min; after the exposure, carrying out neutral erythrocyte toxicity test or MTT (Methyl Thiazolyl Tetrazolium) cytotoxicity test according to a regular method; analyzing a dosage reaction relation between smoke exposure amount and cytotoxicity according to the following formula: full-smoke dilution multiple=( flow speed of smoke + flow speed of clean air)/flow speed of smoke; smoke exposure amount under exposure of 10 mL of a cell suspension solution for 5 min=TPM (Total Particulate Matter) amount of single cigarette/suction times of single cigarette*6 suctions/10 mL; and representing the extent of cytotoxicity by a cell inhibition ratio CI. The detection method provided by the invention has the advantages of simplicity, feasibility, low cost and the capability of providing technical supports for the safety evaluation of cigarette smoke.
Description
Technical field: [0001] The invention relates to a method for detecting the cytotoxicity of a gas-liquid contact cigarette under full smoke exposure, and belongs to the technical field of cigarette smoke safety evaluation. Background technique: [0002] The research on the total smoke exposure method for tobacco toxicology evaluation has caused the world because of its irreplaceable advantages in simulating the human body environment, being able to achieve multiple relevant detection endpoints, and detecting the original whole smoke online in real time. The extensive attention of various tobacco companies and analytical testing laboratories, some tobacco companies have conducted in-depth and systematic research on the exposure method. [0003] At present, there have been patents on the method for external exposure of cigarettes to the whole smoke. Published Patent 1 (application number: 200810230659.0) is a cytotoxicity test method for mainstream cigarette smoke. The main exposure...
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