Method for detecting pear endogenous hormone by using ultra-high performance liquid chromatography electrospray tandem mass spectrum

An ultra-high performance liquid chromatography and tandem mass spectrometry technology is applied in the field of detecting endogenous hormones in pears by ultra-high performance liquid chromatography, electrospray tandem mass spectrometry, and can solve the problems of thermal instability of plant endogenous hormones, restricting development, and complex plant body components. , to achieve the effect of simple and reliable method, efficient extraction and reasonable design

Inactive Publication Date: 2013-10-16
ZHEJIANG UNIV
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Problems solved by technology

GC-MS has become one of the conventional methods for the analysis of plant endogenous hormones due to its advantages of rapid analysis, high sensitivity, and simultaneous qualitative and quantitative analysis. development of
[0006] Compared with GC-MS, ultra-high performance liquid chromatography tandem mass spectrometry has made great progress in speed, sensitivity and resolution. It is widely used in food, traditional Chinese medicine and other fields, but there are few reports on the analysis of plant endogenous hormones. Phase chromatography tandem mass spectrometry has a higher analytical throughput for trace components with complex matrices, but it still needs to be based on reliable analysis samples. Plant endogenous hormones are thermally unstable and plant components are complex. How to quickly and It is still a difficult problem to effectively obtain analysis samples, establish ideal analysis conditions, and reliably perform qualitative and quantitative detection of plant endogenous hormones.

Method used

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  • Method for detecting pear endogenous hormone by using ultra-high performance liquid chromatography electrospray tandem mass spectrum
  • Method for detecting pear endogenous hormone by using ultra-high performance liquid chromatography electrospray tandem mass spectrum
  • Method for detecting pear endogenous hormone by using ultra-high performance liquid chromatography electrospray tandem mass spectrum

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Experimental program
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Effect test

Embodiment 1

[0041] 1. Test equipment and reagents

[0042] Nitrogen Dryer (Tianjin Hengao Technology Co., Ltd.)

[0043] Milli Q ultrapure water device (Millipore, USA)

[0044] Vacuum centrifugal concentrator (Concentrator plus, Eppendorf, Germany)

[0045] Ultra-high performance liquid chromatography mass spectrometry (ACQUITY system followed by tandem Waters Quattro Premier XE Mass Spectrometry detection, Waters, USA)

[0046] Liquid chromatography column ACQUITY BEH C181.7um (2.1×100mm, Waters)

[0047] C18 solid phase extraction cartridge (6mL, waters)

[0048] IAA standard (Sigma, USA); ABA standard (Sigma, USA); t-ZR standard (Sigma, USA); GA 4 Standard product (American sigma company)

[0049]2. Sample pretreatment and standard solution configuration

[0050] (1) Put the plant tissue in a pre-cooled mortar and freeze and grind it with liquid nitrogen;

[0051] (2) Weigh 0.3g of the ground plant material into a 10mL centrifuge tube filled with 3mL extract solution I (extra...

Embodiment 2

[0093] Application of embodiment 2 method

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Abstract

The invention discloses a method for detecting a pear endogenous hormone by using an ultra-high performance liquid chromatography electrospray tandem mass spectrum. The method comprises the following steps of: crushing obtained pear tissues, then adding an extracting solution for extraction, after ending extraction, adding an extraction reagent to a system, and extracting to obtain a crude extract of the endogenous hormone; separating and purifying the crude extract by using a solid-phase extraction column to obtain a detection sample; analyzing a detection sample by using the ultra-high performance liquid chromatography electrospray tandem mass spectrum to obtain the variety and the content of the endogenous hormone in the pear tissues, wherein the extracting solution comprises isopropanol, water and hydrochloric acid, and the volume ratio of the isopropanol, the water and the hydrochloric acid is (1.5-2.5):1:(0.001-0.002), and the extraction agent is dichloromethane. The method has the advantages of high sensitivity, simple pretreatment, high recovery rate, high degree of accuracy and degree of precision and the like and is used for detecting and confirming the endogenous hormones in multiple plant issues.

Description

technical field [0001] The invention relates to the technical field of spectrum analysis, in particular to a method for detecting pear endogenous hormones by using ultra-high performance liquid chromatography electrospray tandem mass spectrometry. Background technique [0002] Phytohormones are a type of organic substances produced by the plant's own metabolism, and move from the production site to the action site. 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 滕元文牛庆丰杨奉霞刘国琴
Owner ZHEJIANG UNIV
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