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Small rat in-vivo microkernel analysis testing method for evaluating cigarette flue gas toxity

A technology of cigarette smoke and detection method, which is applied in the field of micronucleus analysis and detection in mice, can solve the problems of misjudgment of micronucleus, heavy counting workload, and many human factors, and achieves the effects of reliable results and high sensitivity

Inactive Publication Date: 2008-08-27
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For micronucleus detection, the cells are made into smears, fixed and stained, and the number of micronuclei in 1000 multi-stained red blood cells is counted with a microscope. The counting workload is huge, there are many human factors, and it is easy to misjudgment of micronuclei

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Evaluate a domestic flue-cured tobacco brand cigarette. Cigarette smoke was collected through organic phase (ethyl acetate) and inorganic phase (basic culture medium). Mix the organic solvent with the culture medium, and adjust the concentration to 1 cigarette / ml with the cell culture medium. Stored at -80°C, one of the CSCs with the highest LD50 measured by oral toxicity is 13.2 tubes / Kg, and the 1 / 2 LD50 is 6.6 tubes / Kg, which will be purchased in SPF grade NIH-q mice (weight 18-22g) were randomly divided into normal control group, positive control group (cyclophosphamide 50mg / Kg) and cigarette group (because there are many groups with a single dose of 6.6 / Kg), 20 in each group (half male and half male), After 36 hours, take 5 μL of tail vein blood and add 40 μL NaHCO containing heparin (10u / ml) 3 Add 1ml methanol pre-cooled at -80°C to the buffer solution (place a 1.5ml centrifuge tube with 1.0ml analytical pure methanol in ethanol containing 1 / 3 water, put it in a...

Embodiment 2

[0025] Evaluate a domestic blended cigarette. Cigarette smoke was collected through organic phase (ethyl acetate) and inorganic phase (basic culture medium). Mix the organic solvent with the culture medium, and adjust the concentration to 1 cigarette / ml with the cell culture medium. Stored at -80°C, one of the CSCs with the highest LD50 measured by oral toxicity is 13.2 tubes / Kg, and the 1 / 2 LD50 is 6.6 tubes / Kg, which will be purchased in SPF grade NIH-q mice (weight 18-22g) were randomly divided into normal control group, positive control group (cyclophosphamide 50mg / Kg) and each cigarette group (due to the large number of groups, a single dose of 6.6 cigarettes / Kg was used), 20 in each group (half male and half male) , administered separately, 36 hours later, 5 μL of tail vein blood was added to 40 μL NaHCO containing heparin (10u / ml) 3 Add 1ml methanol pre-cooled at -80°C to the buffer solution (place a 1.5ml centrifuge tube with 1.0ml analytical pure methanol in ethanol...

Embodiment 3

[0030]Evaluate a certain foreign cigarette. Cigarette smoke was collected through organic phase (ethyl acetate) and inorganic phase (basic culture medium). Mix the organic solvent with the culture medium, and adjust the concentration to 1 cigarette / ml with the cell culture medium. Stored at -80°C, one of the CSCs with the highest LD50 measured by oral toxicity is 13.2 tubes / Kg, and the 1 / 2 LD50 is 6.6 tubes / Kg, which will be purchased from SPF grade NIH-q mice (weight 18-22g) were randomly divided into normal control group, positive control group (cyclophosphamide 50mg / Kg) and each cigarette group (due to the large number of groups, a single dose of 6.6 cigarettes / Kg was used), 20 in each group (half male and half male) , administered separately, 36 hours later, 5 μL of tail vein blood was added to 40 μL NaHCO containing heparin (10u / ml) 3 Add 1ml methanol pre-cooled at -80°C to the buffer solution (place a 1.5ml centrifuge tube with 1.0ml analytical pure methanol in ethanol...

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Abstract

The invention provides an analysis and detection method for in vivo micronuclei of a mouse for judging the toxicity of cigarette smokes; the invention is characterized in that the invention adopts the in vivo micronucleus detection mode, and can directly measure the micronucleus rate by the antibody fluorescence labeling method after processes of the stomach filling and toxicity contaminating of the mouse, the cell culture, the fixing and the dyeing. The steps of the invention are as follows: (a) the mouse is selected; (b) agglutinators of the cigarette smokes are prepared; (c) the mouse is contaminated with the toxicity of cigarette smoke condensates (CSC); (d) the mouse is bled for culturing, fixing, dyeing and counting; (e) test results are processed and analyzed so as to judge the toxicity of the cigarette smokes. Compared with the prior art, the invention has the advantages of high sensibility and reliable result, and provides a novel detection method for judging the toxicity of the cigarette smokes.

Description

technical field [0001] The invention relates to the field of toxicity evaluation of cigarette smoke condensate, in particular to a method for analyzing and detecting micronucleus in mice to evaluate the toxicity of cigarette smoke, which uses the mouse micronucleus test to evaluate the toxicity of cigarette smoke. It provides a highly sensitive and reliable method for evaluating the toxicity of cigarette smoke. Background technique [0002] In recent years, a lot of research work has been carried out internationally on the hazard assessment of tobacco products and tobacco additives. [0003] In 1999, FDA (Food and Drug Administration) asked IOM (Institute of Medicine, American Academy of Sciences) to develop a set of scientific methods for evaluating the safety of long-term use of drug products or tobacco substitute products (used to reduce rather than eliminate tobacco harm) and effects. Based on the current research results, biomarkers for perception of tobacco products ...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/48G01N21/00
Inventor 谢剑平朱茂祥杨陟华聂聪赵乐彭斌谢复炜刘惠民
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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