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Method for determining total selenium content in selenium-rich proteoglycan by high performance liquid chromatography

A high-performance liquid chromatography and selenium-enriched protein technology, which is applied in measuring devices, material excitation analysis, fluorescence/phosphorescence, etc., can solve problems such as loss and ineffective separation of interfering substances, and avoid matrix interference and multiple transfers Effect

Active Publication Date: 2020-10-30
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Zhou et al. used liquid-liquid dispersion microextraction and reversed-phase C 18 The column high performance liquid chromatography ultraviolet detection method was used to determine the total selenium content in tea leaves, but in the process of applying this method to the determination of the total selenium content in selenoproteoglycans, it was found that the concentration of the extractant in the pretreatment process would cause 4,5-benzene And loss of kohlrabi brain, more importantly common C 18 Cannot effectively separate 4,5-benzokohlrabi selenium from interfering substances in the system

Method used

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  • Method for determining total selenium content in selenium-rich proteoglycan by high performance liquid chromatography
  • Method for determining total selenium content in selenium-rich proteoglycan by high performance liquid chromatography
  • Method for determining total selenium content in selenium-rich proteoglycan by high performance liquid chromatography

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 The drawing of standard curve

[0025] Accurately weigh 50.0 mg of sodium selenite and dissolve it in 500 mL of 0.1 mol / L HCl, and gradually dilute with 0.1 mol / L HCl to form 500.0, 250.0, 100.0, 50.0 and 25.0 µg / L sodium selenite standard working solutions . After step 0005, use RPHPLC-UV-FLD analysis to determine the content of 4,5-benzokohlrabiselenium.

Embodiment 2

[0026] Example 2 National Standard Method Determination of Total Selenium Content

[0027] Accurately weigh 0.5 g~3 g of sample, add 10 mL of HNO3-HClO4 mixed acid (9+1) to digest overnight. After 12 hours, heat and digest on an electric heating plate until the remaining volume is about 2 mL. After cooling, add 5 mL of hydrochloric acid solution (6 mol / L), and continue heating until the remaining volume is about 2 mL. After cooling, dilute to 100 mL with water, measure part of the solution (pH 1.5-2.0, Se content ≤0.4 µg), add 3 mL of hydroxylamine hydrochloride and 2 mL of DAN, shake well, and place in a boiling water bath for 5 min. After cooling, it was extracted with 10 mL cyclohexane, and the fluorescence intensity was measured with a fluorescence photometer under the condition of excitation wavelength of 376 nm and emission wavelength of 520 nm, so as to calculate the selenium content in the sample.

Embodiment 3

[0028] Determination of total selenium content in embodiment 3 seleno-enriched proteoglycans

[0029] Weigh 25 mg selenoproteoglycan sample, add 10 mL HNO 3 -HClO 4 Mixed acid (9ml HNO 3 and 1ml HClO 4 ) for cold digestion overnight; after 12 hours, heat and digest on an electric heating plate to a remaining volume of 2 mL, then add 5 mL of hydrochloric acid solution (6 mol / L) after cooling, continue heating to a remaining volume of 2 mL, and adjust the pH value to 1.5~ 2.0, and dilute to 250 mL with water, take 3 mL of the sample into a 5 mL stoppered centrifuge tube, add 0.5 mL of DAN to it, react in a boiling water bath for 10 minutes, and after cooling in running water, add 400 μL of acetonitrile to it successively and 120 μL of chlorobenzene, shake for 5 minutes, then stand still for 5 minutes, and finally centrifuge at 5000 rpm / min for 5 minutes; pipette 100 μL of chlorobenzene and add 100 μL of acetonitrile to it, mix well and inject the sample. Separation by PFP c...

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Abstract

The invention discloses a method for determining the content of total selenium in selenium-enriched proteoglycan by high performance liquid chromatography. The invention relates to a method for producing 2, 3, 5, 6-tetramethylpiperidine, 2, 3-diaminonaphthalene is used as a chelating agent; 400 microliters of acetonitrile is used as a dispersing agent; the method comprises the following steps: diluting a chelate (4, 5-benzo kohlrabi selenium brain) of selenium (IV) and 2, 3-diaminonaphthalene by acetonitrile, directly separating by using a PFP chromatographic column, eluting by using acetonitrile, measuring the fluorescence intensity by using a fluorescence detector under the conditions that the excitation wavelength is 376 nm and the emission wavelength is 520 nm, and quantifying by usingan external standard method. The method has the characteristics of stable experimental result, less sample transfer, less reagent consumption, high recovery rate and the like.

Description

technical field [0001] The invention relates to a method for measuring the total selenium content in seleno-enriched proteoglycan by high performance liquid chromatography. Background technique [0002] Selenium (Se) is an essential trace element for animal life, has a narrow safety threshold, and its different biological activities depend on different forms of existence. The research on the analysis methods of Se in different matrices has always attracted extensive attention. In addition, selenite and organic selenium compounds are added to the feed as a feed additive. Therefore, it is very necessary to establish a reliable selenium determination method. At present, there are various highly selective analytical techniques for the determination of selenium content, including atomic absorption spectrometry (AAS), atomic emission spectrometry (AES), inductively coupled plasma mass spectrometry (ICP-MS)] and atmospheric pressure chemical ionization mass spectrometry ( APCI-MS...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/36G01N30/74G01N21/64
CPCG01N30/02G01N30/06G01N30/36G01N30/74G01N21/6428G01N2030/062
Inventor 王凤芹汪以真曹进平路则庆程远之
Owner ZHEJIANG UNIV
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