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Method of detecting effect of oral tobacco products upon early apoptosis

A technology for snus tobacco products and tobacco products, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems affecting the test results of neutrophil toxicity method, etc., and achieve the effect of accurate and effective detection

Inactive Publication Date: 2015-06-24
CHINA TOBACCO YUNNAN IND
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Problems solved by technology

This change may affect the test results of the neutrophil toxicity method

Method used

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  • Method of detecting effect of oral tobacco products upon early apoptosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] It is used to detect the effect of Camel brand mint flavored Snus snus products on the early apoptosis rate of cells.

[0029] (1) Sample pretreatment:

[0030] Accurately weigh 1.0g of snus product, add 30mL of serum-free medium, stand in a constant temperature water bath at 37°C for 24 hours before extracting, first filter with qualitative filter paper to remove residue, and then use 0.45μm organic Sterilize by membrane filtration to obtain the liquid to be tested;

[0031] (2) Preparation of single cell suspension:

[0032] Human oral mucosal fibroblast hOMF cells were stored at 37°C, 5% CO 2 , 95%RH (relative air humidity) incubator, remove the incubator when the cell confluence rate is 80-90%, wash twice with phosphate buffer, discard the washing solution, and then add 1-2mL concentration of 0.25 % (w / v) trypsin solution to incubate the monolayer for 1-2 minutes, add DMEM / F12 cell culture medium to suspend to form a single cell suspension;

[0033] (3) Calculat...

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Abstract

The invention discloses a method of detecting effect of oral tobacco products upon early apoptosis. The method includes: pretreating a sample, preparing single-cell suspension, calculating cell concentration of the single-cell suspension, inoculating cells, setting groups, setting dosage, adding a test sample, incubating the test sample, collecting cells, washing the cells, adding resuspended cells, mixing Annexin V-FITC and PI (propidium iodide) dye, allowing reaction, adding binding buffer, detecting the test sample after mixing, and performing detecting with a flow cytometer in one hour to obtain an early apoptosis influence rate. Through effective treatment of the sample, optimal setting of detection dosage and correct selection of target cells, the invention allows the effect of the oral tobacco products upon early apoptosis to be accurately and effectively detected.

Description

technical field [0001] The invention relates to a method for detecting the effect of snus products on early apoptosis of cells, and belongs to the technical field of biological safety evaluation of tobacco and tobacco products. Background technique [0002] Increasingly stringent tobacco regulatory legislation and in-depth research on smoking and health have led tobacco companies to seek new tobacco products with lower risks and no environmental smoke. In 2002, CORESTA, the International Tobacco Scientific Research Cooperation Center, established a working group on toxicological testing of cigarette smoke, and recommended the toxicological testing methods of traditional cigarette smoke. However, the use of snus products is different from that of traditional cigarette inhalation. Snus products are used directly by mouth, and their dissolved substances will enter the digestive system with saliva. During use, no smoke is produced, and the exposure route is significantly differe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02
Inventor 管莹李雪梅夭建华米其利朱洲海高茜唐萍
Owner CHINA TOBACCO YUNNAN IND
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