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Preparation method of phosphorylated serine imprinted polymer monolithic micro-column

A technology of phosphorylated serine and imprinted polymer, applied in chemical instruments and methods, separation methods, other chemical processes, etc., can solve the problem of low ionization efficiency of P-Ser mass spectrometry analysis, high-abundance non-phosphorylated amino acids, large interference, and inability to Quantify P-Ser and other issues to achieve the effect of improving analytical selectivity, improving sensitivity, and not easy to swell

Active Publication Date: 2019-11-22
HUBEI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Phosphorylated serine (P-Ser) comes from the breakdown of phosphorylated proteins or cell phospholipids, and the increase in its content is related to the severity of the disease. Although the relationship between P-Ser and the disease cannot be quantified at present, studies have shown that P-Ser Related to liver and kidney dysfunction, cancer and other diseases, abnormal P-Ser levels usually indicate the existence of diseases, so the analysis and detection of P-Ser has important medical significance
Mass spectrometry (MS) is rich in chemical information and can be analyzed quickly and with multiple components. It is an effective technique for the analysis of biological samples, but the matrix of biological samples is complex, the interference of high-abundance non-phosphorylated amino acids is large, the content of P-Ser is low, and P- The ionization efficiency of Ser mass spectrometry is very low

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  • Preparation method of phosphorylated serine imprinted polymer monolithic micro-column
  • Preparation method of phosphorylated serine imprinted polymer monolithic micro-column
  • Preparation method of phosphorylated serine imprinted polymer monolithic micro-column

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Embodiment 1

[0027] combine figure 1 The schematic flow chart of the present invention for preparing the phosphorylated serine imprinted polymer monolithic microcolumn is shown, and the preparation method of the phosphorylated serine imprinted polymer monolithic microcolumn specifically includes steps:

[0028] (a) Take P-Ser (1.1mmol, 0.2g) and methanol solution of tetrabutylammonium hydroxide (0.8mol / L, 2.75ml) and mix evenly, add to 5ml of methanol, stir evenly, and then add 1.55ml dropwise Water yields the reaction product.

[0029] (b) Take 240 μL of the reaction product, add 250 μL of methanol, 8.1 mg of acrylamide, sonicate for 1 hour, add 108 μL of EGDMA and 5 mg of AIBN for 10 minutes to obtain a prepolymer, pipette 100 μL of the prepolymer into the pipette tip, and pipette One end of the pipette tip was sealed with methanol, and the other end of the pipette tip was sealed with silica gel. The prepolymer was subjected to aqueous phase in-situ polymerization in the sealed pipette ...

Embodiment 2

[0035] Using the phosphorylated serine imprinted polymer monolithic microcolumn obtained in the embodiment to separate and enrich phosphorylated serine, and establish a method for HPLC-MS / MS separation and analysis of phosphorylated serine in pig liver samples with high sensitivity and high selectivity, including steps: The pig liver sample solution was injected into the phosphorylated serine imprinted polymer monolithic microcolumn, phosphorylated serine was selectively adsorbed on the phosphorylated serine imprinted polymer monolithic microcolumn, and the phosphorylated serine imprinted polymer monolithic microcolumn was eluted with eluent. phosphorylated serine in the column to obtain an eluate, and finally use HPLC-MS / MS technology to analyze the content of phosphorylated serine in the eluate. The specific process of injecting the pig liver sample solution into the monolithic microcolumn of phosphorylated serine imprinted polymer is that the pipette tip containing the monol...

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Abstract

The invention relates to the technical field of monolithic column preparation, in particular to a preparation method of a phosphorylated serine imprinted polymer monolithic micro-column. The preparation method of the phosphorylated serine imprinted polymer monolithic micro-column comprises the following steps: (a) mixing phosphorylated serine, tetrabutylammonium hydroxide, methanol and water, andperforming a reaction to obtain a product; (b) with the reaction product as a template compound in a sealed reaction container, adding methanol, a functional monomer, a cross-linking agent and an initiator to carry out water-phase in-situ polymerization reaction, and washing the products after the reaction is finished, so as to obtain the phosphorylated serine imprinted polymer monolithic micro-column. The obtained phosphorylated serine imprinted polymer monolithic micro-column can realize specific adsorption of phosphorylated serine.

Description

technical field [0001] The invention relates to the technical field of monolithic column preparation, in particular to a preparation method of phosphorylated serine imprinted polymer monolithic microcolumn. Background technique [0002] Amino acid phosphorylation plays a key regulatory role in many biochemical processes. Phosphorylated serine (P-Ser) comes from the breakdown of phosphorylated proteins or cell phospholipids, and the increase in its content is related to the severity of the disease. Although the relationship between P-Ser and the disease cannot be quantified at present, studies have shown that P-Ser Related to liver and kidney dysfunction, cancer and other diseases, abnormal P-Ser levels usually indicate the existence of diseases, so the analysis and detection of P-Ser has important medical significance. Mass spectrometry (MS) is rich in chemical information and can be analyzed quickly and with multiple components. It is an effective technique for the analysi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/22B01J20/285C08F220/56C08F222/14C08J9/26G01N30/02G01N30/06G01N30/36G01N30/72
CPCB01J20/285B01D15/22C08F220/56C08F222/1006C08J9/26G01N30/02G01N30/06G01N30/36G01N30/72G01N2030/027G01N2030/062
Inventor 陈怀侠周胜兰孙源艾佑宏党雪平
Owner HUBEI UNIV