Determination method of trehalose-6-phosphate content in tomato by ultra-high performance liquid chromatography-tandem mass spectrometry
An ultra-high performance liquid chromatography and tandem mass spectrometry technology is applied in the field of ultra-high performance liquid chromatography tandem mass spectrometry detection of trehalose-6-phosphate content in tomato, which can solve the problems of low sensitivity, difficulty, and low recovery rate, and improve the sensitivity , Simple steps and high accuracy
Active Publication Date: 2022-03-18
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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[0004] Due to the wide variety of compounds and complex structures in plants, the content of trehalose-6-phosphate is extremely low, which makes the extraction and quantitative analysis of trehalose-6-phosphate in plants more difficult, and thus hinders the study of plant algae. The Scientific Study of Sugar-6-Phosphate
At present, the extraction of plant trehalose-6-phosphate is carried out by solid phase extraction, which is not only cumbersome to process, but also has a low recovery rate, and the cost of the experiment is high, which is not conducive to the rapid processing of samples in batches.
The determination method using liquid chromatography alone has the defects of many interferences and low sensitivity
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[0040] In order to better understand the invention, the present invention will be described in detail below in connection with specific embodiments.
[0041] Ultra-high-performance liquid chromatographic tandem spectrum detection method of trehalose-6-phosphate content, including the following steps:
[0042] 1. Prepare standard working solutions and perform super high efficiency liquid chromatography tandem syringe test:
[0043] Weigh 0.5 mg of trehalose-6-phosphate standard in 10 ml volumetric flask, dissolved with acetonitrile with a volume fraction of 80%, and disposable as a standard stock solution; the standard stock solution is 80% acetonitrile The graded dilution was complemented to the trehalose-6-phosphate concentration of 25 μg / ml, 12.5 μg / ml, 6.25 μg / ml, 3.125 μg / ml, 1.562.5 μg / ml, 0.78125 μg / mL, 0.390625μg / ml. Standard solution.
[0044] Experimental conditions for ultra-efficient liquid chromatographic tandem mass spectrometry are as follows:
[0045] ...
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The invention discloses an ultra-high performance liquid chromatography tandem mass spectrometry detection method for the content of trehalose-6-phosphate in tomato, comprising the following steps: (1) preparing a standard working solution and performing ultra-high performance liquid chromatography tandem mass spectrometry detection; (2) according to The standard spectrogram that step (1) obtains makes the concentration of standard working solution and the chromatographic peak area relationship curve; (3) the pretreatment of tomato sample obtains the sample solution of trehalose-6-phosphate content to be detected; (4) super High-performance liquid chromatography tandem mass spectrometry detection; (5) Determination of the content of trehalose-6-phosphate in the sample: determine the chromatographic peak of trehalose-6-phosphate by the retention time of the sample spectrum obtained in step (4), and The chromatographic peak area of trehalose-6-phosphate is compared with the concentration and chromatographic peak area relationship curve of the standard working solution in step (2), and the content of trehalose-6-phosphate in the sample solution is determined by the interpolation method.
Description
Technical field [0001] The present invention belongs to the quantitative detection of trehalose-6-phosphoric acid in plant samples, and specifically, it is specifically related to ultra-high performance liquid chromatographic tandem mass spectrum detection of trehalose-6-phosphoric acid content. Background technique [0002] Tomatoes are widely cultivated in the world, which is a fruit and vegetable that is loved by people. Tomato production is an important part of my country's modern agriculture. With the continuous improvement of technology, as well as the improvement of people's living standards, high-yield, high-quality tomatoes have become people's production goals. The improvement of tomato quality and yield is closely related to the production and distribution of tomato photosynthetic products. Sugar is the main form of photosynthetic products, not only provides energy supply for plants, but also serves as a signal molecule induction plant energy and transmits to plant met...
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IPC IPC(8): G01N30/02G01N30/06G01N30/14G01N30/54
Inventor 蒋卫杰李强余宏军
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI