Purification method of plant endogenous brassinolide based on boron affinity solid phase extraction

A technology of brassinosteroids and boron affinity, applied in the field of analytical chemistry, can solve the problems of affecting derivatization efficiency, lower sensitivity, low recovery rate, etc., and achieve the effects of easy operation, reduced sample usage, and simple method

Active Publication Date: 2018-04-20
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the following problems usually exist in the process of purifying BRs with boron-affinity materials: (1) It needs to be used in conjunction with solid-phase extraction materials of other mechanisms to remove impurities from BRs: for example, plant extracts generally need to be reversed-phase Or normal-phase solid-phase extraction materials to remove impurities, and then interact with boron-affinity materials; (2) plant extracts need to remove water before interacting with boron-affinity materials: such as extraction of plant materials after freez...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Purification method of plant endogenous brassinolide based on boron affinity solid phase extraction
  • Purification method of plant endogenous brassinolide based on boron affinity solid phase extraction
  • Purification method of plant endogenous brassinolide based on boron affinity solid phase extraction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1, Determination of Brassinolide in Paddy Nipponbare Flowering Stage Ear

[0045] Weigh 10 mg of the ground plant sample (wild-type rice Nipponbare flowering stage panicle) and put it into a 2 mL centrifuge tube, add 1 mL of an aqueous methanol solution with a concentration of 60% extraction solvent by mass, then add 25-100 pg of a stable isotope internal standard, and mix well . Ultrasonic extraction at room temperature for 1 h. The supernatant was obtained by centrifugation, and the supernatant was loaded onto a boron affinity solid-phase extraction column (1CC, 100mg, BondElut PBA, Agilent). 5% methanol solution of formic acid, methanol and water rinse the solid phase extraction column in sequence, then use 5% hydrogen peroxide to oxidize the complex formed by the combination of BRs and boron affinity material, and then use 5% ammonia solution to hydrolyze the oxidation product, After rinsing with water to remove the generated boric acid, BRs was finally elut...

Embodiment 2

[0046] Example 2, Determination of Brassinolide in Rice Nipponbare Leaves

[0047] Weigh 10 mg of the ground plant sample (rice Nipponbare leaf) into a 2 mL centrifuge tube, add 1 mL of 80% methanol, then add 25-100 pg of stable isotope internal standard, and mix well. Ultrasonic extraction at room temperature for 1 h. Centrifuge to get the supernatant, and interact the supernatant with a boron affinity solid-phase extraction column (1CC, 100mg, Bond Elut PBA, Agilent), and use a formic acid aqueous solution with a mass percent concentration of 5%, and a formic acid mass percent concentration of 5% methanol solution of formic acid, methanol and water rinse the solid phase extraction column in sequence, then use 10% hydrogen peroxide to oxidize the complex formed by the combination of BRs and boron affinity material, and then use 10% ammonia solution to hydrolyze the oxidation product, After rinsing with water to remove the generated boric acid, BRs was finally eluted in rever...

Embodiment 3

[0048] Embodiment 3, the mensuration of brassinolide in rice Nipponbare stem

[0049] Weigh 10 mg of the ground plant sample (rice Nipponbare stem) into a 2 mL centrifuge tube, add 1 mL of methanol, then add 25-100 pg of stable isotope internal standard, and mix well. Ultrasonic extraction at room temperature for 1 h. Centrifuge to take the supernatant, and interact the supernatant with 5 mg of boron-affinity material (Benzeneboronic acid, polymer-supported, boronic acid group content 2.6-3.2mmol / g, Alfa Aesar) dispersed in solid phase extraction, expressed in mass percent Concentration is 5% formic acid aqueous solution, formic acid mass percent concentration is 5% formic acid methanol solution, methanol and water rinse the solid phase extraction column successively, then use 5% ammonia solution containing 5% hydrogen peroxide to oxidatively hydrolyze BRs and The complex formed by the combination of the boron affinity material was washed with water to remove the generated bo...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Particle sizeaaaaaaaaaa
Login to view more

Abstract

The invention discloses a purification method of plant endogenous brassinolide based on boron affinity solid phase extraction. According to the method, the plant endogenous brassinolide is extracted by a solvent, an extract liquid directly acts on a boron affinity material, the plant endogenous brassinolide is adsorbed on the surface of the material, matrix impurities are removed through leaching,and then the brassinolide adsorbed on the boron affinity material is eluted through an oxidation-hydrolysis-reverse phase elution strategy. The method is easy to operate, good in selectivity, high inrecovery rate, and thorough in removal of the matrix interference, can be used for high-sensitivity and quantitative analysis of trace brassinolide in plant materials, and has wide application prospects in plant genetics, crop science and other fields.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and relates to a method for purifying plant endogenous brassinosteroids based on boron affinity solid-phase extraction. Background technique [0002] Brassinosteroids (BRs) are an important class of plant hormones, which participate in the regulation of plant growth and development at extremely low concentrations, including regulation of flowering, pollen development, organ differentiation, etc.; in addition to direct regulation of growth and development In addition, BRs are also involved in plant responses to abiotic and biotic stresses, including salt, drought, and extreme temperatures. Accurate quantitative analysis of endogenous BRs is of great significance for studying the molecular mechanism of BRs action. However, due to the extremely low content of BRs in plants and the complex plant matrix, the establishment of a high-sensitivity quantitative analysis method for BRs has always been a ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N30/06
CPCG01N30/06G01N2030/062
Inventor 辛培勇褚金芳李兵兵闫吉军
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap