Method for determining acute toxicity of direct dyes by using saccharomyces cerevisiae
A technology of Saccharomyces cerevisiae, acute toxicity, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., to achieve the effect of low determination cost
Inactive Publication Date: 2012-02-08
WUHAN TEXTILE UNIV
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[0013] The technical problem to be solved by the present invention is: to develop a novel rapid detection method for the acute toxicity of microorganisms, which can integrate the interaction of various toxic substances, determine the relationship between the mass concentration of dyes and their printing and dyeing wastewater and their biological effects. Carry out risk assessment to provide scientific basis for ensuring the safe production and use of dyes, formulating discharge standards for printing and dyeing wastewater treatment, early warning of environmental pollution, monitoring and comprehensive evaluation of water quality
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[0025] Determination of EC of direct light fast turquoise blue GB (also known as tinny blue GL, direct light fast turquoise blue GL, direct light fast sapphire blue; Lionol Blue GB) 50 .
[0026] 1) Preparation of Saccharomyces cerevisiae suspension: In the ultra-clean workbench, use a sterile toothpick to collect a single colony of Saccharomyces cerevisiae cultured on the plate and transfer it to a sterilized Eppendorf tube labeled ①, add 1000 μl of sterile pure water to suspend , take 200 μl of the suspension and add it to a 5 ml sterile test tube, add 1.8 ml sterile purified water, measure the OD on a spectrophotometer 600 =0.248, it is known that the cell density of the suspension in tube ① is 0.248÷1.4×10 8 ×10=1.7714×10 8 pcs / ml; take 56.4 μl from tube ① into tube ②, add 944 μl sterile purified water to make 1×10 7 cells / ml of bacterial suspension; dilute the bacterial suspension in tube ② 1000 times, that is, the cell density is about 1.0×10 4 cells / ml of the bacter...
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The invention relates to the field of textile safety and dye production, in particular to a novel method for rapidly detecting toxicity of direct dyes. The method is characterized in that: after a system consisting of 20 microliters of saccharomyces cerevisiae suspension with the density of 1.0 * 10<4> cells / ml, 130 microliters of a series of dye solution with concentration to be detected and 110 microliters of 2.2-time yeast peptone dextrose (YPD) liquid medium in a cover-connecting centrifugal tube is subjected to shaking culture for 6 to 7.5 hours on a constant temperature shaker at the temperature of 30 DEG at 150 r / min, the cell number is determined by using a hand-held cell counter based on a resistance method cell counting principle or a dull colony counting method, and EC50 of the dyes is calculated. A 96-well cell culture plate (or an elisa plate) replaces the cover-connecting centrifugal tube, and each 96-well plate can determine four kinds of dyes. The method can be used for detecting the comprehensive acute toxicity of various water soluble inorganic matters, water soluble organic matters or microorganisms.
Description
technical field [0001] The invention relates to the fields of textile safety and dye production, and is a method for evaluating the acute toxicity of direct dyes to biological cells. Background technique [0002] Direct dyes have a strong affinity for fibers, and can dye cellulose fibers without the help of mordants, so they account for a large number of dyes for cellulose fibers. These dye molecules contain sulfonic acid groups or carboxyl groups. The commercial product is made as a sodium salt, so it is soluble in water. The structure of direct dyes has the characteristics of linearity, coplanarity, and long conjugated system. The chemical structure is mainly polyazo dyes with sulfonic acid groups such as disazo and trisazo. It is mainly used in the dyeing of cotton yarn, knitted fabrics, viscose fiber and silk, and also in the dyeing of fur and paper. [0003] The raw materials of most direct dyes are aromatic amines, which are a class of carcinogens. Therefore, in the...
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Inventor 谭之杨
Owner WUHAN TEXTILE UNIV