Neutral erythrocyte toxicity testing method for biological evaluation of cigarette tipping paper

A technology of cigarette tipping paper and biological evaluation, applied in the field of tobacco, to achieve the effect of practical method and suitable dose range

Inactive Publication Date: 2011-08-17
YUNNAN RES INST OF TOBACCO SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there have been no public reports at home and abroad on the neutral erythrocyte toxicity test method for the biolo...

Method used

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  • Neutral erythrocyte toxicity testing method for biological evaluation of cigarette tipping paper
  • Neutral erythrocyte toxicity testing method for biological evaluation of cigarette tipping paper
  • Neutral erythrocyte toxicity testing method for biological evaluation of cigarette tipping paper

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0024] Example 1 Evaluation of HH Cigarette Tipping Paper

[0025] 1 Experimental part

[0026] 1.1 Main instruments and reagents

[0027] Series II CO 2 Incubator (ThermoForma Company); inverted microscope (Olympus Company); microplate reader (Bio-Rad Company); 96-well cell culture plate (Corning Company); cell culture flask (Corning Company).

[0028] DMEM medium (Dulbecco's modification of Eagle's medium, Solarbio); fetal bovine serum (Clark); phosphate buffer (CMF-PBS, Solarbio); trypsin (Solarbio); neutral red dye ( Sigma Company); penicillin-streptomycin (Solarbio Company); formaldehyde; ethanol; glacial acetic acid; organic solvents were all analytically pure.

[0029] The cell culture medium was prepared by adding 10% (v / v) fetal calf serum and 100 units / mL penicillin-streptomycin to DMEM medium. The neutral red solution was prepared in DMEM medium with a concentration of 100 μg / mL. The acidic ethanol solution is prepared by volume ratio, glacial acetic acid: abso...

example 2

[0043] Example 2 Evaluation of HD cigarette tipping paper

[0044]Basically the same as Example 1. The differences are:

[0045] Measure HD cigarette tipping paper 300cm×6cm (equivalent to the amount of tipping paper for 250 cigarettes), cut into thin strips of about 1cm×6cm, and place it in a 250mL conical flask with a ground stopper. Inject 150mL of prepared artificial saliva into the Erlenmeyer flask equipped with HD cigarette tipping paper, so that the tipping paper strip is completely immersed (the ratio of tipping paper area / extraction solution is 24cm 2 / mL, which is equivalent to extracting the tipping paper of 3.3 cigarettes with 1mL extracting solution), and cover the stopper. Simulate the actual exposure conditions, set the extraction temperature to 37±1°C, place the Erlenmeyer flask in a water bath shaker at 37°C for 1 hour of vibration extraction (oscillation speed 200r / min), and prepare HD cigarette tipping paper sample stock solution.

[0046] It can be known...

example 3

[0050] Example 3 Evaluation of YY cigarette tipping paper

[0051] Basically the same as Example 1. The differences are:

[0052] Measure YY cigarette tipping paper 120cm×6cm (equivalent to the amount of tipping paper for 100 cigarettes), cut it into thin strips of about 1cm×6cm, and place it in a 50mL conical flask with a ground stopper. Inject 20mL of prepared artificial saliva into the Erlenmeyer flask with YY cigarette tipping paper, so that the tipping paper strip is completely immersed (the ratio of tipping paper area / extraction solution is 36cm 2 / mL, which is equivalent to extracting the tipping paper of 5 cigarettes with 1mL extracting solution), and cover the stopper. Simulate the actual exposure conditions, set the extraction temperature to 37±1°C, place the Erlenmeyer flask in a water bath shaker at 37±1°C for 180 minutes (oscillation speed 200r / min), and prepare the tipping paper sample stock solution.

[0053] It can be known from Table 3 that the stock soluti...

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Abstract

The invention relates to a neutral erythrocyte toxicity testing method for biological evaluation of a piece of cigarette tipping paper and belongs to the technical field of tobaccos. The method concretely comprises the following steps of: taking artificial saliva as tipping paper leach liquor; measuring and taking the tipping paper with a certain area, shearing to a strip with a length and a width of 1cm and 6cm and placing in a 50-250ml conical flask with a ground glass stopper for later use; pouring the leach liquor into the conical flask according to a proportion of the tipping paper area to the leach liquor of 24-36cm<2>/ml and putting back the stopper; placing the conical flask in a water-bath vibrator at a temperature of 36-38 DEG C, vibrating and leaching for 5-180min and preparing a sample stock solution; carrying out a neutral erythrocyte toxicity test, wherein doses of the sample stock solution per ml of nutrient solution successively are 1mg, 5mg, 10mg, 25mg, 50mg, 100mg, 200mg, 300mg, 400mg, 500mg; and evaluating the security of the tipping paper according to the result of the neutral erythrocyte toxicity test. The method provided by the invention has the advantages that the actual situation of the contact between the tipping paper and a mouth is effectively simulated to prepare a sample; the dose range of the sample stock solution for the neutral erythrocyte toxicity test is suitable; and the method is practical, simple and convenient.

Description

Technical field: [0001] The invention relates to a neutrophil toxicity test method for biological evaluation of cigarette tipping paper, which belongs to the technical field of tobacco. Background technique: [0002] Although cigarette tipping paper does not participate in combustion, it is directly in contact with the consumer's mouth. At present, the attention on cigarette tipping paper is mainly focused on the specification of its physical indicators, microbial indicators, heavy metals and quality indicators [1-2]. With the continuous deepening of harm reduction and tar reduction, new types of cigarette tipping papers continue to appear, and it is necessary to carry out in-depth research on the safety of cigarette tipping papers, so as to provide basic data and theoretical support for the supervision of cigarette materials. [0003] Although the sample preparation method used for the biological evaluation of cigarette tipping paper is not the core of safety evaluation wo...

Claims

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Application Information

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IPC IPC(8): G01N33/48
Inventor 夭建华魏玉玲杨叶昆李雪梅米其利唐萍黄海涛缪明明
Owner YUNNAN RES INST OF TOBACCO SCI
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