Method for determining endogenous hormones in peanut seeds

Abstract

The invention provides a method for determining endogenous hormones in peanut seeds. The method comprises the following steps: grinding the peanut seeds with liquid nitrogen into peanut seed powder and leaching the peanut seed powder twice with methanol; extracting with chloroform for three times, adding crosslinked polyvinylpyrrolidone into an aqueous phase, oscillating to remove phenolic substance and centrifuging; adjusting the pH to be less than 7 with hydrochloric acid, extracting for three times with ethyl acetate, mixing upper ester phases, and carrying out blow-drying with nitrogen to obtain a first residue; adjusting the pH of the lower aqueous phase to be greater than 7, extracting for three times with water-saturated n-butanol, mixing upper oil phases, carrying out blow-drying with nitrogen to obtain a second residue; respectively dissolving the first residue and the second residue in a mixed solution of methanol and water, analyzing by virtue of high-performance liquid chromatography and well separating endogenous hormones through gradient elution of high-performance liquid chromatography. The problems of overlapping of peak pattern, excessive denseness, tailing and the like of chromatogram are avoided and four hormones are extracted under different pH conditions so that various hormones can be extracted to the greatest extent.

Description

technical field [0001] The invention relates to the field of analytical chemistry, in particular to a method for measuring endogenous hormones in peanut kernels. Background technique [0002] Phytohormones are a type of organic substances produced by the plant's own metabolism. They move from the site of production to the site of action. They are trace substances that have obvious physiological effects at very low concentrations. They are also called plant natural hormones or plant endogenous hormones. [0003] Plant hormones are closely related to cell division and elongation, tissue and organ differentiation, flowering and fruiting, maturation and senescence, dormancy and germination, and in vitro tissue culture, and regulate plant growth, development and differentiation separately or in coordination with each other. The flexibility and diversity of this regulation can be achieved by changing the concentration and ratio of exogenous hormones or synthetic plant growth regul...

AI Technical Summary

Problems solved by technology

[0005] In the prior art, high-performance liquid chromatography is used to measure the content of endogenous hormones in seeds, and polyvinylpyrrolidone is generally used to remove phenols in peanut seeds, but polyvinylpyrrolidone is water-soluble, which affects the extraction effect; in acidic conditions 3-indole acetic acid, abscisic acid and gibberellin can be extracted to the maximum extent, while cytokinin can only be extracted under alkaline conditions. The existing technology extracts the four hormones together, which greatly reduces the The amount of hormone extracted leads to inaccurate final test results; the existing technology adopts isocratic elution, which leads to problems such as peak overlap, excessive density, and tailing of the spectrum, and endogenous hormones cannot be well separated; at the same time, there are The operation is complicated, the cost is high, only one sample can be processed at a time, time-consuming, and the loss of the analyte during the extraction process is large, etc.

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