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Magnetic microsphere for extracting and purifying paraquat metabolite, preparation method, kit and extraction method

A magnetic microsphere and extraction method technology, applied in the field of biological analysis, can solve the problems of unfriendly liquid chromatography-mass spectrometry, less quantitative analysis research, less detection objects, etc., achieves good ability to remove matrix interference, and improves extraction Purification speed, the effect of rapid solid-liquid phase magnetic separation

Pending Publication Date: 2022-03-01
NINGBO MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there are few studies on the quantitative analysis of paraquat metabolites in poisoned biological samples. The paper by Ma Hongjuan et al. (Ma Hongjuan, et al. metabolites (monoquat and paraquat monopyridone (MP)), and the metabolites of paraquat also include 4-carboxy-1-methyl pyridinium ion (MINA) and paraquat dipyridone (DP), so this literature has fewer detection objects and cannot fully reflect the paraquat Metabolism in the human body
On the other hand, in the sample pretreatment process in this paper, pure water dilution and acetonitrile precipitation are used simply, which is difficult to remove the high concentration of inorganic salts and phospholipid components that cause matrix interference in blood samples, and the accurate quantification of paraquat metabolites Analysis is very unfavorable, and high concentration of inorganic salts and phospholipids are extremely unfriendly to liquid chromatography-mass spectrometry, which can easily lead to blockage of liquid chromatography sampling system and ion source electrospray needle of mass spectrometer, and high concentration of phospholipids Components are easy to contaminate the liquid chromatographic column, resulting in poor accuracy and repeatability of test results

Method used

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  • Magnetic microsphere for extracting and purifying paraquat metabolite, preparation method, kit and extraction method
  • Magnetic microsphere for extracting and purifying paraquat metabolite, preparation method, kit and extraction method
  • Magnetic microsphere for extracting and purifying paraquat metabolite, preparation method, kit and extraction method

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Embodiment 1

[0044] The preparation of embodiment 1 magnetic microsphere

[0045] This example prepares a magnetic microsphere for extracting and purifying paraquat metabolites, which can extract and purify four paraquat metabolites in plasma: monoquat, paraquat monopyr idone (MP), 4-carboxy-1 -methyl pyridinium ion (MINA) and paraquat dipyridone (DP).

[0046] The preparation method comprises the following steps:

[0047] S11, Fe 3 o 4 Synthesis of microspheres

[0048] Using solvothermal method, weigh 0.3-0.5g sodium citrate and 0.65-0.85g FeCl 3 ·6H 2 O in a 250mL flask, add 80mL ethylene glycol, mechanically stir and dissolve at room temperature for 30min; then add 5g sodium acetate and continue stirring for 30min; then, put the above mixed solution into a 100mL autoclave, react at 200°C for 8h, and cool to room temperature; then wash with pure water and ethanol three times respectively, and vacuum dry overnight at 60°C.

[0049] S12, Fe 3 o 4 @SiO 2 -Synthesis of MPS microsp...

Embodiment 2

[0056] Example 2 Kit

[0057] This embodiment provides a kit for extracting and purifying paraquat metabolites, the adsorption material used in the kit is the magnetic microspheres prepared in Example 1.

[0058] The kit includes EDTA anticoagulant tubes, disposable syringes, standard series of working solutions, absorbent material dispersions, 2mL polypropylene centrifuge tubes, eluent and reconstitution solutions. The solute of the adsorption material dispersion is magnetic microspheres, the solvent is a mixed phosphate buffer solution, the pH is 6.5-9.5, the concentration of the magnetic microspheres is 5-20 mg / mL, and the eluent is 1% to 5% formic acid acetonitrile solution , the complex solution is acetonitrile aqueous solution (volume ratio is 1:1 ~ 1:5).

[0059] The kit in this embodiment can be combined with a magnetic frame and a small vortex instrument to realize the on-site sampling, processing and storage of plasma samples. Afterwards, the pretreated sample solut...

Embodiment 3

[0060] Example 3 Extraction of Paraquat Metabolites in Plasma Samples

[0061] This embodiment is a method for extracting paraquat metabolites in plasma samples using the kit of embodiment 3, comprising the following steps:

[0062] S21. Accurately pipette 200 μL of plasma sample into a 2 mL polypropylene centrifuge tube, add 1.2 mL of adsorption material dispersion, vortex for 5-10 min, then place on a magnetic stand for magnetic separation for 10 s, and discard the supernatant.

[0063] S22. Wash with 1mL water and 1mL methanol respectively, magnetically separate for 10s, discard the supernatant; add 1.5mL 2% formic acid acetonitrile solution to elute, vortex extract for 4-10min, magnetically separate for 10s, collect the supernatant, 40 Blow dry with nitrogen at °C.

[0064] S23. Add 200 μL of acetonitrile-water solution (1:1, v / v) for redissolution, and inject and analyze by LC-MS.

[0065] LC-MS instrument conditions are as follows:

[0066] (1) Liquid phase conditions...

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Abstract

The invention discloses a magnetic microsphere for extracting and purifying paraquat metabolite, a preparation method, a kit and an extraction method.According to the kit, amphiphilic carboxyl functionalized magnetic microspheres with a core-shell structure serve as an adsorption material, and a plasma sample can be efficiently and rapidly purified and treated; the matrix interference effect in the paraquat metabolite detection process is effectively reduced, and the concentrations of four metabolites of trace paraquat in a plasma sample of a poisoned patient are rapidly and accurately detected.

Description

technical field [0001] The invention relates to the technical field of biological analysis, in particular to a magnetic microsphere for extracting and purifying paraquat metabolites, a preparation method, a kit and an extraction method. Background technique [0002] Paraquat is a fast-killing herbicide, which is extremely toxic to the human body. The mortality rate of oral poisoning is as high as 50%. . At present, although the production and use of paraquat has been banned in my country, due to the difficulty of management, there are still unscrupulous traders operating and selling it, especially the phenomenon that the product name of the pesticide is inconsistent with the actual composition. For example, paraquat was illegally added to diquat and glufosinate-ammonium pesticides to improve the weed-killing effect, which resulted in many cases of poisoning by mistaken ingestion, and caused extremely serious adverse effects on society. In recent years, the incidence of acu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 潘胜东尚春庆陈晓红金米聪
Owner NINGBO MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION
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