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Morphological analysis method for selenium in selenium-rich protein polysaccharide

A technology for seleno-enriched protein and speciation analysis, which is applied to the analysis of materials, material separation, measurement devices, etc., can solve the problems of not being able to meet the needs of rapid detection, and achieve the effects of environmental protection, low efficiency, and high recovery rate

Inactive Publication Date: 2017-02-01
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the general enzymatic hydrolysis process is long, and most of them need to be incubated at 37°C for 24 hours (Moreno, 2001). This process often cannot meet the needs of rapid detection, and structural differences between selenium compounds are prone to occur during this process. Transformation (Capelo, 2004)

Method used

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  • Morphological analysis method for selenium in selenium-rich protein polysaccharide
  • Morphological analysis method for selenium in selenium-rich protein polysaccharide
  • Morphological analysis method for selenium in selenium-rich protein polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 The drawing of standard curve

[0026] Accurately weigh 20.0 mg of SeCys2, add it into water containing dilute acid to make a 200 ug / mL standard stock solution, and then gradually dilute to 0.50, 0.30, 0.20, 0.10 and 0.05 ug / mL SeCys2 standard working solution. Accurately weigh 20.0 mg of SeCys, add it into water containing dilute acid to make a 200 ug / mL standard stock solution, and then gradually dilute it into 5.0, 4.0, 2.5, 1.0, 0.8, and 0.5 ug / mL SeCys standard working solutions. Accurately weigh 25.0 mg Na 2 SeO 3 Dissolve in water to make 100ug / mL Na 2 SeO 3 Standard stock solution, then gradually diluted to 0.50, 0.30, 0.20, 0.05 and 0.025 ug / mLNa 2 SeO 3 Standard working solution. According to the conditions in Table 1, apply HPLC-IPC-MS analysis to make a standard curve.

[0027]

Embodiment 2

[0028] Example 2 SeCys2, SeCys and Na 2 SeO 3 Assay

[0029] Accurately weigh 5 portions of 25 mg selenium-enriched proteoglycans into a 5 mL centrifuge tube, add 3 mg XIV protease, and then add 3 mL of water, sonicate at 37°C for 30 min, then centrifuge for 30 min at 10,000 rpm / min , collected the supernatant, and filtered it through a 0.45 μm aqueous phase filter membrane, and then applied HPLC-ICP-MS to analyze the selenium compounds in the enzymatic hydrolysis supernatant: SeCys2, SeCys and Na 2 SeO 3 Analyze and calculate SeCys2, SeCys and Na respectively according to the standard curve 2 SeO 3 content.

Embodiment 3

[0030] SeCys2, SeCys and Na in embodiment 3 selenoproteoglycan 2 SeO 3 Determination of the recovery rate of

[0031] Accurately weigh 3 parts of 25 mg selenoproteoglycan into a 5 mL centrifuge tube, add 3 mg XIV protease, and then add 1.5 mL of 0.30 ug / mL SeCys2, 8.0 ug / mL SeCys and 0.50 ug / mL SeCys2 Na 2 SeO 3 and supplemented with water to 3 mL, ultrasonicated at 37°C for 30 min, then centrifuged for 30 min at a speed of 10,000 rpm / min, collected the supernatant, and applied HPLC-ICP-MS to enzymatically hydrolyze the selenium compounds in the supernatant: SeCys2, SeCys and Na 2 SeO 3Analyze and calculate SeCys2, SeCys and Na respectively according to the standard curve 2 SeO 3 The content of, in conjunction with embodiment 2, calculate SeCys2, SeCys and Na 2 SeO 3 recovery rate.

[0032] According to the result and discussion that above embodiment 1-3 obtains:

[0033] 1. Drawing of standard curve

[0034] For SeCys2, SeCys and Na 2 SeO 3 To make a standard cu...

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Abstract

The invention discloses a morphological analysis method for selenium in selenium-rich protein polysaccharide. The method includes the steps that protease XIV is added into selenium-rich protein polysaccharide, ultrasound-assisted enzymolysis is carried out, and high performance liquid chromatography-inductively coupled plasma-massspectrometry (HPLC-ICP-MS) is adopted to analyze selenium compounds in enzymolysis supernate, wherein the selenium compounds include selenocystine (SeCys2), selenocysteine (SeCys) and sodium selenite (Na2SeO3). The method meets the requirements for environmental friendliness and economy, enzymolysis lasting for 24 h or a longer time is not needed, and the recovery rate is higher than that of a common method.

Description

technical field [0001] The invention relates to a method for analyzing the form of selenium in selenium-enriched proteoglycan. Background technique [0002] Selenium is the active center of many enzymes such as glutathione peroxidase (Jagtap, 2016), and at the same time, it helps to fight cancer and prevent aging (Axley, 1991). However, the range between selenium toxicity and requirement is very narrow, and the biological function of selenium is closely related to its form (Suhajda, 2000). Inorganic selenium mainly has two forms of selenate and selenite, while organic selenium has three forms of selenoamino acid, seleno small peptide and selenoprotein (Maseko, 2013). Compared with inorganic selenium, organic selenium has higher biological activity and lower toxicity, and is more easily absorbed by the body (Kieliszek, 2013; Kouba, 2014). Therefore, it is necessary to establish a speciation analysis method for selenium. [0003] In order not to change the form of selenium ...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 王凤芹汪以真张林路则庆
Owner ZHEJIANG UNIV
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