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Method for determining grass endogenous hormones

A technology of endogenous hormones and hormones, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that are not conducive to the accurate determination of plant endogenous hormone content, complex operation, low recovery rate, etc., to shorten the sample loading time, Good repeatability and the effect of improving the recovery rate

Inactive Publication Date: 2019-04-09
陆欣瑜
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

This method is not only complicated to operate, but also the recovery rate is quite low (about 80%), which is not conducive to accurately determine the content of plant endogenous hormones.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] (1) Select more representative turfgrass (Poa annua and Lolium: separate extraction and detection, both using the method of this embodiment) as samples. Cut 0.3g of fresh leaves, add liquid nitrogen and grind until pulverized.

[0041] (2) Add 3 mL of sodium phosphate buffer solution (0.05M, pH=7.0, containing 0.02% sodium diethyldithiocarbamate; keep the leaf weight in g: extract volume mL=1:10), transfer to a 10 mL centrifuge tube After shaking at 4°C for 1 h, adjust the pH to about 2.6 with 1M hydrochloric acid, then add 3 mL of dichloromethane, and shake at 4°C for 1 h.

[0042] Note: Plant endogenous hormones are very unstable, so ensure that the shaking process is always maintained at 4°C.

[0043] (3) Take out the centrifuge tube, centrifuge the extract obtained in the previous step at 10,000 rpm for 10 min, remove the supernatant (about 3 mL) and transfer it to another centrifuge tube. Add 3 mL of dichloromethane to the residue, continue shaking at 4°C for 1 h...

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Abstract

The invention relates to a method for determining grass endogenous hormones, comprising the following steps: 1) carrying out twice extraction on a sodium phosphate buffer solution for lawn grass and dichloromethane, carrying out twice centrifugation, and then, blow-drying with nitrogen to obtain lawn grass endogenous hormones; 2) dissolving the lawn grass endogenous hormones obtained in the step 1) with methanol, decolorizing and degreasing by using a solid-phase extraction column, taking eluant, then, carrying out vacuum pumping, and dissolving with a mixed solution of methanol and acetic acid; and 3) taking methanol and acetic acid as a mobile phase, and using high-performance liquid chromatography to determine the lawn grass endogenous hormones treated in the step 2) to obtain the content of the lawn grass endogenous hormones. According to the method in the invention, an appropriate endogenous hormones extraction method and chromatographic conditions are selected to accurately determine the content of the lawn grass endogenous hormones. Compared with the prior art, the method has the advantages of being high in recovery rate, high in extraction efficiency, high in accuracy rateand good in repeatability.

Description

technical field [0001] The invention relates to the field of measuring plant endogenous hormones, in particular to a method for measuring grass endogenous hormones. Background technique [0002] Since high performance liquid chromatography was used to analyze plant endogenous hormones in the 1970s, due to its high sensitivity, selectivity, good repeatability and fast analysis speed, it has been used in the four major types of plant hormones and growth regulators except ethylene. The field of drug research has been continuously developed. The focus of the quantitative determination of plant endogenous hormones by high performance liquid chromatography is to choose the appropriate extraction method and chromatographic conditions for endogenous hormones. However, due to the complexity and particularity of biological endogenous hormones, the separation and extraction of target components is an extremely critical step, and is an important factor that directly affects its precisi...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 陆欣瑜
Owner 陆欣瑜
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