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Solid-phase extracting column and application thereof

A technology of solid-phase extraction column and glass column, which is applied in material separation, instruments, and analytical materials, etc., can solve problems such as inability to quickly and efficiently separate and enrich, affect sample processing efficiency, and cannot be eluted, and achieve strong application value , high accuracy, easy loading effect

Active Publication Date: 2012-10-17
SHANGHAI UNIV OF T C M
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 3) If the target compound forms a covalent bond with the adsorbent, it may not be eluted due to dead adsorption with the adsorbent during elution;
[0007] 4) The extraction flow rate is low during use, which affects the sample processing efficiency and requires a certain pressure from the outside world;
[0008] 5) Unable to simultaneously enrich compounds with similar molecular weights
In order to achieve higher adsorption efficiency and reduce sample loss, the traditional granular packing has small particle size, but the small particle size will cause high column pressure and long adsorption time, so it cannot be used for fast and efficient separation. and enrichment

Method used

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  • Solid-phase extracting column and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1: Using the solid phase extraction column of the present invention to process angiotensin oligopeptides in serum

[0030] Take a self-made hollow glass column with a length of 15 cm and an inner diameter of 1.0 cm, fill it with Sephadex LH-20 gel, elute with methanol continuously, fill it up, and use it as a stationary phase.

[0031] Take 1ml of serum, add 4ml of methanol, vortex and mix immediately, and centrifuge at 4°C for 15 minutes; take 4.5ml of supernatant, blow dry with nitrogen, redissolve with 0.5ml of methanol solution, and centrifuge again at 4°C for 15 minutes; The supernatant was slowly loaded to the gel solid-phase extraction column, continuously eluted with methanol, and the 3-9ml eluted fractions were collected.

[0032] After concentrating the methanol to dryness, reconstitute with 150 μL of a solution containing 20 mmol / L ammonium formate and 0.1% (V:V) formic acid and acetonitrile at a volume ratio of 88:12, and centrifuge at 4°C (20,000...

Embodiment 2

[0039] Embodiment 2: Using the solid phase extraction column of the present invention to process phospholipids in serum

[0040] Take a self-made hollow glass column with a length of 15 cm and an inner diameter of 1.0 cm, fill it with Sephadex LH-20 gel, elute with methanol continuously, fill it up, and use it as a stationary phase.

[0041] Take 1ml of serum, add 4ml of methanol, vortex and mix immediately, and centrifuge at 4°C for 15 minutes; take 4.5ml of supernatant, blow dry with nitrogen, redissolve with 0.5ml of methanol solution, and centrifuge again at 4°C for 15 minutes; The supernatant was slowly loaded to the gel solid-phase extraction column, eluted continuously with methanol, and the 6-10ml eluted fractions were collected.

[0042] After concentrating methanol to dryness, reconstitute with 150 μL of a solution containing 10 mmol / L ammonium acetate and 0.1% (V:V) formic acid and acetonitrile at a volume ratio of 60:40, and centrifuge at 4°C (20,000 rpm) Get supe...

Embodiment 3

[0046]Example 3: Using the solid phase extraction column of the present invention to process bile acid compounds in liver homogenate

[0047] Take a self-made hollow glass column with a length of 15 cm and an inner diameter of 1.0 cm, fill it with Sephadex LH-20 gel, elute with methanol continuously, fill it up, and use it as a stationary phase.

[0048] Take 0.2mg of the liver, add 1ml of methanol to homogenate, add 4ml of methanol to the homogenate, vortex and mix immediately, and centrifuge at 4°C for 15 minutes; take 4.5ml of the supernatant, dry it with nitrogen, and reconstitute with 0.5ml of methanol solution. Centrifuge again at 4°C for 15 minutes; then take the supernatant and slowly load it to the gel solid phase extraction column, elute with methanol continuously, and collect the 6-10ml eluted fractions.

[0049] After concentrating methanol to dryness, reconstitute with 150 μL of a solution containing 5 mmol / L ammonium acetate and 0.1% (V:V) formic acid and methano...

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Abstract

The invention discloses a solid-phase extracting column, comprising a glass column and a filler. The filler is hydroxypropyl sephadex LH-20. The solid-phase extracting column can be used for pretreatment of biological samples, in particular used for simultaneous concentration and analysis detection of middle and small molecules in serum, urine and tissue homogenate and the molecular weight of middle and small molecules is close. The solid-phase extracting column in the invention can suppress interference effectively, ensure high sensitivity, good specificity and high accuracy of analysis of analytes and has excellent guiding significance to organism metabonomics research and detection of disease. Furthermore the solid-phase extracting column in the invention can be used repeatedly and hardly exist the phenomenon of regeneration. The extracting column is easy in filling, simple in operation. The column has stronger application value in analysis of biological samples.

Description

technical field [0001] The invention relates to a solid phase extraction column and its application, in particular to a gel solid phase extraction column and its application in biological sample pretreatment. Background technique [0002] Due to the complex composition of biological samples and serious matrix interference, the analysis and detection of endogenous substances in biological samples has always been concerned by analysts. Generally, biological sample pretreatment methods include organic solvent precipitation (OSP), liquid-liquid extraction (LLE), solid phase extraction (SPE), solid phase microextraction (SPME) and so on. Among them, solid-phase extraction is widely used for its simple operation, high recovery rate, good repeatability, and environmental protection. [0003] Solid-phase extraction can be divided into four types: reversed-phase solid-phase extraction, normal-phase solid-phase extraction, ion-exchange solid-phase extraction and adsorption solid-phas...

Claims

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Application Information

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IPC IPC(8): B01D15/08G01N30/06
Inventor 杨莉祁萌王峥涛耿放
Owner SHANGHAI UNIV OF T C M
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