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Solid-phase extraction column, preparation method thereof and lactoferrin detection method

A solid-phase extraction column and detection method technology, applied in the field of biochemical detection, can solve the problems of cumbersome separation and purification, high cost, etc., and achieve the effect of ensuring sufficient nutrition and good application prospects

Active Publication Date: 2020-11-13
SHENZHEN POLYTECHNIC +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a solid-phase extraction column and its preparation method and a lactoferrin detection method, aiming to solve the technical problems of cumbersome separation and purification of samples for lactoferrin determination and high cost

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  • Solid-phase extraction column, preparation method thereof and lactoferrin detection method
  • Solid-phase extraction column, preparation method thereof and lactoferrin detection method
  • Solid-phase extraction column, preparation method thereof and lactoferrin detection method

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preparation example Construction

[0028] On the one hand, the embodiment of the present invention provides a kind of preparation method of solid-phase extraction column, comprises the following steps:

[0029] S01: Provide silane coupling agents, chelating agents, carrier materials and metal ion precursors;

[0030] S02: combining the silane coupling agent and the chelating agent to obtain a cross-linked complex;

[0031] S03: dissolving the cross-linked complex, the carrier material and the metal ion precursor in a solvent to react to obtain a mixed filler;

[0032] S04: placing the mixed filler in an extraction column mold for solidification treatment to obtain a solid phase extraction column.

[0033] In the preparation method of the solid phase extraction column provided in the embodiment of the present invention, the silane coupling agent and the chelating agent are reacted to obtain a cross-linked compound, and the carrier material and the metal ion precursor are dissolved in a solvent for reaction to o...

Embodiment 1

[0077] Embodiment 1 solid phase extraction column filler synthesis and packing process

[0078] (1) Weigh 4.24g IDA and 2.82g NaOH respectively in a 500mL Erlenmeyer flask, add 50mL ultrapure water for ultrasonic dissolution, slowly add 1.4mL 3-Gly dropwise in an ice-water bath at 0°C, and then add 1.4mL 3-Gly dropwise at 65°C at 200~ Stir at a speed of 300rpm for 6 hours, react the common silane reagent with the chelating ligand to form a silane coupling agent-chelating ligand with a dicarboxyl functional group, after the reaction is completed, naturally cool to room temperature, and store in a refrigerator at 4°C.

[0079] (2) Take out the reacted material in step (1) from the refrigerator, slowly add 1.6mL of 3-Gly dropwise, stir at 65°C at a speed of 200-300rpm for 6h, after the reaction is completed, cool naturally to room temperature, and then adjust the pH with concentrated hydrochloric acid To 6.8 ~ 7.2, stored in a 4 ℃ refrigerator.

[0080] (3) Weigh 50g of polystyr...

Embodiment 2

[0083] Example 2 Lactoferrin Detection

[0084] Sample pretreatment

[0085] Accurately weigh 1g (accurate to 0.01g) sample of infant formula milk powder in a 50mL beaker and wait for extraction. First add 5 mL of warm disodium hydrogen phosphate solution (temperature not exceeding 50°C) to the sample, stir to dissolve the sample completely, and transfer all the sample into a 50 mL centrifuge tube. Then add 5 mL of disodium hydrogen phosphate solution to the original beaker, transfer to a centrifuge tube and combine the two liquids. Shake with a vortex shaker for 0.5 min to mix, and then centrifuge at 10,000 r / min for 10 min at 4°C. Transfer the centrifuged supernatant to another 50mL centrifuge tube, then add 5mL disodium hydrogen phosphate solution to the original centrifuge tube, repeat the vortex and centrifugation steps once, and combine the supernatant obtained from the two centrifuges Used as a sample extract.

[0086] Sample testing

[0087] Purification: Open the...

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Abstract

The invention belongs to the technical field of biochemical detection, and particularly relates to a solid-phase extraction column, a preparation method thereof and a lactoferrin detection method. Thepreparation method of the solid-phase extraction column comprises the following steps: providing a silane coupling agent, a chelating agent, a carrier material and a metal ion precursor; combining the silane coupling agent with the chelating agent to obtain a cross-linked compound; dissolving the cross-linked compound, the carrier material and the metal ion precursor in a solvent to carry out a reaction to obtain a mixed filler; and putting the mixed filler into an extraction column mold, and carrying out a curing treatment to obtain the solid-phase extraction column. The prepared solid-phaseextraction column can well adsorb and purify lactoferrin by utilizing the principle of combining metal chelating affinity chromatography and solid-phase extraction, can accurately, simply, conveniently and quickly determine lactoferrin in infant formula milk powder by combining with liquid chromatography subsequently, and has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of biochemical detection, and in particular relates to a solid-phase extraction column, a preparation method thereof and a lactoferrin detection method. Background technique [0002] Lactoferrin (LF) is one of the main proteins in breast milk. In order to make the nutritional content of infant formula milk powder closer to that of breast milk, lactoferrin is a highly safe nutritional additive that can be ingested by infants. , is widely used in infant formula milk powder. At present, there are many varieties of infant formula milk powder on the market, but whether the lactoferrin content in the milk powder is in line with the label range, or whether it really contains lactoferrin is still unknown. Nowadays, there is no national standard for the determination of lactoferrin in infant formula milk powder at home and abroad, so establishing an accurate and efficient detection method for lactoferrin in infant f...

Claims

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Application Information

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IPC IPC(8): B01J20/281B01J20/286B01J20/28B01J20/26B01J20/30B01D15/38B01D15/22B01D15/10G01N30/02G01N30/08G01N30/14
CPCB01D15/10B01D15/22B01D15/3804B01J20/22B01J20/261B01J20/28021B01J20/281B01J20/286B01J2220/4806B01J2220/4812G01N30/02G01N30/08G01N30/14G01N2030/062
Inventor 张毅岳振峰罗耀江婷坤温景岚
Owner SHENZHEN POLYTECHNIC
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