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Method for detecting mutagenicity of total particulate matter of cigarette smoke

A technology for mutagenicity and cigarette smoke, applied in biochemical equipment and methods, microbiological measurement/inspection, etc., can solve the problem of inappropriate dose range and inability to reflect the dose-response relationship of total particulate matter in cigarette smoke, etc. problem, to achieve a good dose-response relationship, obvious bacterial reverse mutation, and shorten the test time

Inactive Publication Date: 2010-10-06
YUNNAN RES INST OF TOBACCO SCI
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Problems solved by technology

For total particulate matter in cigarette smoke, this dose range is inappropriate and too wide to reflect the dose-response relationship of total particulate matter in cigarette smoke

Method used

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  • Method for detecting mutagenicity of total particulate matter of cigarette smoke

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Embodiment Construction

[0024] 1. Preparation of total particulate matter (TPM) in cigarette smoke

[0025] The cigarettes were balanced in a constant temperature and humidity chamber for 48 hours, and cigarettes with consistent weight and draw resistance were selected. According to the national standard method, 20 cigarettes were smoked on the smoking machine, and the Cambridge filter was soaked in DMSO at 10 mgTPM / ml, ultrasonically extracted for 20 minutes, finally filtered with filter paper, packed into cryopreservation tubes, and stored in a -70°C ultra-low temperature refrigerator in spare.

[0026] 2. Identification of strains

[0027] The identification of bacterial strains in this laboratory complies with the operating procedure document [1] of food toxicology testing standards for bacterial strain identification, and the identification is used for experiments after passing the identification. Two commercial strains, MOLTOX AMES ASSAY STDisc TA98 and TA100, were used as strains.

[0028] ...

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Abstract

The invention relates to a method for detecting the mutagenicity of the total particulate matter of cigarette smoke, and belongs to the technical field of tobacco and cigarette smoke safety evaluation. The method is improved on the basis of an Ames test plate incorporation method; and the improvement points are that: a, the amount of histidine is 20 to 25 mg / ml, the amount of top layers is 2.5 ml, and the holding temperature of the top layer is 43 DEG C; b, the dosage range of the mutagenicity of the total particulate matter of the cigarette smoke is 25 to 500 mu g / vessel; and c, the positive substance of a strain TA98 is 2-aminofluorene of which the adopted dosage is 1 to 10 mu g / vessel; and the positive substance of a strain TA100 is sodium azide of which the adopted dosage is 0.15 to 1.5 mu g / vessel. The method has the advantages of simplified operation steps, reduced test period, consistent effect, obvious bacterial reverse mutation, reduced positive substance dosage, proper dosage range of selected total particulate matter of the cigarette smoke, and capability of detecting the mutagenicity of the total particulate matter of the cigarette smoke.

Description

Technical field: [0001] The invention relates to a method for detecting the mutagenicity of total particulate matter in cigarette smoke, and belongs to the technical field of safety evaluation of tobacco and cigarette smoke. Background technique: [0002] The Ames test is an experimental method for detecting short-term DNA mutations caused by exogenous compounds. Its basic principle is to use the characteristics of exogenous compounds to induce the reverse mutation of histidine-deficient Salmonella typhimurium to detect the mutagenic ability of exogenous compounds. It is an internationally recognized routine method for the detection of chemical mutagens, and it is also one of the main methods for the detection of food toxicology and environmental chemical mutagens in the past two decades. This test can sensitively detect the potential genetic hazards induced by total particulate matter in cigarette smoke, and the results are relatively reliable, so it has become one of the p...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
Inventor 夭建华倪红梅黄海涛李雪梅缪明明米其利
Owner YUNNAN RES INST OF TOBACCO SCI
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