High-throughput continuous flow analysis method for total plant alkaloid in tobacco gene editing material

Abstract

The invention discloses a high-throughput continuous flow analysis method for total plant alkaloid in a tobacco gene editing material. The method includes the steps of: A. accurately weighing 0.25g ofa sample containing total plant alkaloid and putting the sample in 25mL sample tubes, and putting the sample weighed sample injection tubes into a sample injection disc of a continuous flow analyzerautomatic sample injector in order till full; B. adding 25mL of deionized water into each sample tube respectively with a pipetting gun, then inserting inner sleeve tubes with filter sieve plates having a pore size of 0.45microm at the bottoms into the sample tubes in order to prevent splashing of the solution during oscillation extraction, then putting the sample disc on an oscillator and conducting oscillation extraction; and C. pressing the inner sleeve tubes downward at the end of extraction, letting the well extracted sample solution enter the inner sleeve tubes through the sieve plates,then putting the sample disc on the automatic sample injector of the continuous flow analyzer, inserting sample injection syringes of the automatic sample injector only into the supernatant in the inner sleeve tubes each time to suck filtered sample, and then transferring the filtered sample into the continuous flow analyzer for analysis.

Description

technical field [0001] The invention belongs to the field of tobacco chemistry research, and in particular relates to a high-throughput analysis method suitable for detecting the content of total alkaloids 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] The total plant alkaloids of tobacco refers to all the alkaloids in tobacco. The most important tobacco alkaloids are nicotine, which accounts for more than 95% of the total amount of tobacco alkaloids, followed by demethylnicotine (nornicotine), an...

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.

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