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Method for determining form of selenium in selenium-rich tea trees and application

A determination method and a technology of morphology, applied in the field of selenium speciation determination in selenium-enriched tea trees, can solve the problems of inability to accurately evaluate the composition and content of selenium in selenium-enriched tea trees, unsuitable tea trees, and complicated detection, so as to improve sample extraction rate and price Inexpensive, high extraction rate effect

Pending Publication Date: 2021-06-15
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The existing commonly used selenium speciation detection method is adapted from "a method for the determination of selenium" (patent number: 201310658482.5), specifically by measuring the content of total selenium and inorganic selenium in the sample, and then using the subtraction method to obtain the organic selenium in the sample. This method cannot accurately evaluate the composition and content of each form of selenium in selenium-enriched tea trees
In "A method for the determination of selenomethionine in selenium-enriched yeast" (patent number: 201110177717.X), chemical reagents such as enzymes are used to separate and extract selenomethionine in yeast, and the content of selenomethionine is determined by HPLC-FLD method. But only one kind of organic selenium can be detected
In addition, the existing selenium speciation detection methods are mainly aimed at fungi, water samples, etc., and the detection of tea tree as a polysaccharide and polyphenol plant is more complicated than other plants and easy-to-hydrolyze samples. These existing methods are not applicable to tea tree

Method used

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  • Method for determining form of selenium in selenium-rich tea trees and application
  • Method for determining form of selenium in selenium-rich tea trees and application
  • Method for determining form of selenium in selenium-rich tea trees and application

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preparation example Construction

[0071] (2) Preparation of sample solution: Weigh 0.2 g of the dried sample into a centrifuge tube, add 20 mmol / L Tris-HCl buffer containing 20 mg of pronase E, and place it in an ultrasonic cleaner at 37 °C for 60 min in a water bath. , Centrifuge at 10000r / min for 20min, collect the supernatant and pass it through a 0.45μm filter membrane for later use.

[0072] In step S104, the reference conditions for analyzing the liquid chromatography atomic fluorescence spectrometer provided in the embodiment of the present invention include:

[0073] (1) Liquid chromatography separation conditions

[0074] Mobile phase: Weigh 5.28g of diammonium hydrogen phosphate and dissolve it in 1000mL of water, adjust the pH to 6.0 with formic acid, filter with suction and ultrasonically degas for 10min, and set aside; chromatographic column: anion exchange column Hamilton PRP X100 (250mm×4mm); flow rate: 1.0mL / min; injection volume: 100μL;

[0075] (2) Atomic fluorescence instrument conditions ...

Embodiment 1

[0087] (1) Sample pretreatment

[0088] ①Sample pretreatment: Clean the selenium-enriched tea leaves, freeze-dry them in a freeze-drying machine at -40°C, and grind the dried samples into powder for later use;

[0089] ② Preparation of sample solution: Weigh 0.2 g of the dried sample into a centrifuge tube, add 20 mmol / L Tris-HCl buffer containing 20 mg of pronase E, and place it in an ultrasonic cleaner at 37 °C for 60 min in a water bath for 10000 r Centrifuge at / min for 20 min, collect the supernatant and pass it through a 0.45 μm filter membrane for later use.

[0090] (2) Analysis reference conditions of liquid chromatography atomic fluorescence spectrometer

[0091] ① Liquid chromatography separation conditions

[0092] Mobile phase: Weigh 5.28g of diammonium hydrogen phosphate and dissolve it in 1000mL of water, adjust the pH to 6.0 with formic acid, filter with suction and ultrasonically degas for 10min, and set aside; chromatographic column: anion exchange column H...

Embodiment 2

[0110] (1) Sample pretreatment

[0111] ①Sample pretreatment: Clean the roots of selenium-enriched tea trees, freeze-dry them in a freeze-drying machine at -40°C, and grind the dried samples into powder for later use;

[0112] ② Preparation of sample solution: Weigh 0.2 g of the dried sample into a centrifuge tube, add 20 mmol / L Tris-HCl buffer containing 20 mg of pronase E, and place it in an ultrasonic cleaner at 37 °C for 60 min in a water bath for 10000 r Centrifuge at / min for 20 min, collect the supernatant and pass it through a 0.45 μm filter membrane for later use.

[0113] (2) Analysis reference conditions of liquid chromatography atomic fluorescence spectrometer

[0114] ① Liquid chromatography separation conditions

[0115] Mobile phase: Weigh 5.28g of diammonium hydrogen phosphate and dissolve it in 1000mL of water, adjust the pH to 6.0 with formic acid, filter with suction and ultrasonically degas for 10min, and set aside; chromatographic column: anion exchange ...

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Abstract

The invention belongs to the technical field of organic selenium content testing, and discloses a method for determining the form of selenium in selenium-rich tea trees and application. The method is characterized by comprising the following steps: preparing a KBH4 solution, a KI solution, an HCl solution and a mixed standard working solution; pretreating a sample and preparing a sample solution; analyzing the reference conditions by using a liquid chromatography-atomic fluorescence spectrometer; performing qualitative analysis on the forms of selenium in the mixed standard working solution and the sample solution; and quantitatively determining the corresponding peak area in the chromatogram of the sample solution according to the standard curve. According to the method, the forms of two kinds of inorganic selenium and three kinds of organic selenium in a selenium-rich tea tree tissue sample can be simply and quickly determined, the stability of the selenium forms is ensured by adopting a freeze drying technology and a diammonium hydrogen phosphate-formic acid mobile phase system, and meanwhile, a Tris-HCl buffer solution and a pronase E ultrasonic extraction system are adopted, so that the sample extraction rate is improved to the greatest extent on the premise of ensuring the stability of the selenium form. The composition and content of the selenium form in a selenium-rich tea tree can be truly reflected.

Description

technical field [0001] The invention belongs to the technical field of organic selenium content testing, and in particular relates to a method and application for the determination of selenium form in a selenium-enriched tea tree. Background technique [0002] At present, selenium is an essential nutrient element that cannot be synthesized by the human body. Its biological functions mainly include: improving human immunity, anti-oxidation, delaying aging, protecting and repairing cells, etc. Tea tree is a non-selenium plant but has a strong ability to enrich selenium. Different varieties of tea trees have different selenium-enriching abilities, and the selenium content of different parts of the same variety of tea trees is also different. The effective selenium content in the soil is the main factor affecting the selenium content of tea trees. The same variety of tea trees grow in different soils, and the selenium content in fresh tea leaves can vary by up to 18 times. The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/74
CPCG01N30/02G01N30/06G01N30/74
Inventor 王伟东黄慧宇柏昊高童唐子贻余有本
Owner NORTHWEST A & F UNIV
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