High-flux continuous flow analysis method for water-soluble sugar in tobacco gene editing material

Abstract

The invention discloses a high-flux continuous flow analysis method for water-soluble sugar in a tobacco gene editing material. The high-flux continuous flow analysis method comprises the following steps: A, accurately weighing 0.25g of a sample containing the water-soluble sugar and putting the sample into a 25mL sample tube; putting sample feeding tubes filled with the weighed sample on a samplefeeding disc of an automatic sample feeding device of a continuous flow analyzer in sequence until the sample feeding disc is fully filled with the sample feeding tubes; B, adding 25mL of a 5 percentacetic acid solution into each sample tube by utilizing a pipette; then inserting inner sleeves with filtering sieve plates with the pore diameter of 0.45mu m at the bottom in sequence, so as to prevent the solution from being splashed in an oscillation and extraction process; after inserting the inner sleeves, putting the sample disc with the inserted inner sleeves onto an oscillator and carrying out oscillation and extraction; C, after finishing the extraction, pressing the inner sleeves and introducing an extracted solution into the inner sleeves through the filtering sieve plates; then putting the sample disc on the automatic sample feeding device of the continuous flow analyzer and only inserting a sample feeding needle of the automatic sample feeding device into supernatant in the inner sleeve each time to suck a filtered sample; then transferring the filtered sample into the continuous flow analyzer and analyzing.

Description

technical field [0001] The invention belongs to the field of tobacco chemistry research, and specifically relates to a high-throughput analysis method suitable for detecting the content of water-soluble sugar in tobacco gene editing materials. Background technique [0002] With the advancement of the tobacco industry's "Tobacco Genome Project Major Project-Based Genome Editing Technology Factory Breeding Program", all gene-edited materials will be screened through chemical composition analysis, and the compatibility test of tobacco leaves and cigarette formula will be passed. Verification, to determine the varieties with strong industrial usability, has become a key technical content urgently needed by the tobacco industry. [0003] Water-soluble sugar is an important conventional chemical component in tobacco and an important indicator for tobacco quality evaluation. Currently, continuous flow analysis is commonly used in the detection of water-soluble sugars in tobacco. ...

AI Technical Summary

Problems solved by technology

During the sample pretreatment process, the sample needs to be transferred several times, which is not convenient enough and affects the efficiency of the detection; and the reliability of the detection results may be affected by the introduction of impurities and the careless spilling of the extract during the transfer process.

On the other hand, the tobacco sample contains a large amount of brown-yellow colored substances, the sample extract is yellow, and the absorption wavelength of the color product of reducing sugar and p-hydroxybenzoic acid hydrazide is close. The inconsistency of the background absorption of each sample will interfere with the measurement results

[0005] Factory breeding for gene editing will produce at least tens of thousands of editing materials. At present, the daily sample processing capacity of water-soluble sugar sample analysis is only about 60. It takes a long time to complete the routine chemical composition analysis of tens of thousands of materials, and the lag of the analysis results will be reduced. It will affect the progress of the entire factory breeding, so there is an urgent need for accurate and reliable high-throughput rapid analysis methods in the process of gene editing breeding

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