Ultra-high performance liquid chromatography-tandem mass spectrometry method (UPLC-MS/MS) for detecting endogenous hormone in tubers of Chinese yams

Abstract

The invention discloses an ultra-high performance liquid chromatography-tandem mass spectrometry method (UPLC-MS / MS) for detecting endogenous hormone in tubers of Chinese yams. The method comprises the steps of carrying out dehydration and freeze-drying on fresh samples of the tubers of the Chinese yams by a freezing dryer, crushing and then feeding a solvent for extracting to obtain crude extract liquid; concentrating and drying the obtained crude extract liquid, redissolving the crude extract liquid with phosphate buffer solution, freezing at the temperature of -40 DEG C, unfreezing and then centrifuging to remove impurities; carrying out ethyl acetate extraction for three times, and collecting the ester phase; adjusting the pH of water phase to 2.8-3.5, carrying out ethyl acetate extraction again for three times, collecting and merging the ester phase; concentrating, drying by distillation, and then redissolving the product with methyl alcohol to obtain the detected sample; analyzing the detected sample by the UPLC-MS / MS. According to the method, the pretreatment process is rapid and simple, the hormone extraction and recovery rate is high, and the UPLC-MS / MS is low in limit of detection, high in accuracy and good in linear relation; furthermore, the method is capable of effectively extracting and quantitatively detecting seven hormones in five types of hormones with little content in the tubers of the Chinese yams.

Description

technical field [0001] The invention belongs to the technical field of detection, and relates to an ultra-high performance liquid chromatography-tandem mass spectrometry method for detecting endogenous hormones in yam tubers. Background technique [0002] Yam is a traditional medicinal and edible plant. It is a tuber of the Dioscoreaceae plant. It has the effects of nourishing the spleen and stomach, nourishing the lungs and kidneys. In-depth study of yam tubers can provide a theoretical basis for high-yield and high-quality production of yam. [0003] Plant endogenous hormones are naturally synthesized organic compounds in plants, which play important regulatory roles in the process of growth, development and response to the environment. Among them, gibberellins (GAs), cytokinins (such as ZT) , auxin (such as IAA), abscisic acid (ABA), and jasmonic acid (JA) are four very important endogenous hormones in plants. Studies have shown that plant hormones play an important rol...

AI Technical Summary

Problems solved by technology

However, these methods have some deficiencies in the accurate identification and quantification of plant endogenous hormones. For example, enzyme-linked immunosorbent assay (ELISA) cannot quantitatively analyze multiple plant hormones at the same time, and has unavoidable cross-reactions. The accuracy and reproducibility are poor; chromatography is only qualitative based on retention time, and in complex plant extracts, it may give wrong conclusions because the retention time of different substances is the same or similar, and ultraviolet and fluorescence detectors cannot meet ultra-low Requirements for qualitative and quantitative analysis of plant hormones; capillary electrophoresis method has poor reproducibility; mass spectrometry is currently the most reliable method for detecting plant endogenous hormones, but GC-MS method requires complex derivatization treatment to improve the volatility and Sensitivity, cumbersome operation, and high temperature at the gas phase inlet can lead to degradation of some thermally unstable compounds, which limit the application of GC-MS method

Images