Mutagenesis detection method for urine of rats after smoke inhalation and exposure

Abstract

The invention discloses a mutagenesis detection method of urine after exposure of smoke inhalation in rats. The mutagenesis detection method includes six steps of urine enzymolysis, chromatographic column passing and extraction, elution, concentration, an Ames test micro fluctuation method and result determination, wherein sodium acetate-acetate buffer is adopted in the urine enzymolysis to regulate and control a pH environment, pretreated by methanol and water in sequence when chromatographic column passing is carried out, washed in the deionized water, eluted with methanol, after concentration, the Ames test micro fluctuation method is used for detecting and determining results. According to the mutagenesis detection method, the amount of urine detection required by a traditional Ames test can be significantly reduced, the detection sensitivity is improved, and the mutagenesis detection method is especially suitable for the biological detection of rat smoke exposure models.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a mutagenic detection method for urine after rat smoke inhalation exposure. Background technique [0002] Cigarette smoke is a complex aerosol containing more than 6,000 compounds, 69 of which are identified as human carcinogens by IARC. Polycyclic aromatic hydrocarbons, various alkylating agents, benzene series, aromatic amines and other mutagenic substances in cigarette smoke, after entering the human body or experimental animals, these substances are metabolized in the body, and most of them are prototypes, metabolites or conjugates and other forms are excreted from the kidneys, so it is of great significance to use short-term mutagenicity tests to detect the mutagenic effect of urine. [0003] There are great differences in smoke exposure among the population. Therefore, Sprague Dawley rats were used as test subjects to detect the mutagenicity of rat urine after different cycles...

AI Technical Summary

Problems solved by technology

However, in the current prior art, the urine pretreatment method for the urine mutagenicity test requires a urine volume of 50-100 mL or more. Therefore, the existing method is only applicable to the mutagenicity of human urine. However, the amount of urine in rats is very small, and the existing methods cannot effectively evaluate its mutagenicity

[0004] Ames et al. used the traditional plate incorporation method to detect the mutagenicity of urine with Salmonella typhimurium in the presence of microsomal enzyme system, because the test substance limit per plate of this method is 0.1 mL, and the group in urine Amino acid can also interfere with the experimental results, so this traditional method can only detect high concentrations of mutagenic substances, and cannot be applied to urine with low concentrations of mutagenic substances