Preparation method of test sample for detecting arachidonic acid-like isomer based on magnetic beads

By using a magnetic bead-based sample preparation method, the challenges of separating and quantifying arachidonic acid isomers have been solved, achieving efficient separation and high-sensitivity detection, and promoting the development of metabolomics research towards high-throughput automation.

CN121476480APending Publication Date: 2026-02-06THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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Patent Information

Application Number
CN202511482027.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-10-16
Publication Date
2026-02-06

AI Technical Summary

Technical Problem

Existing technologies struggle to efficiently separate and quantify arachidonic acid isomers, especially due to their similar molecular weights, chemical structures, and low abundance, leading to difficulties in qualitative and quantitative analysis, low pretreatment efficiency, and a high misjudgment rate.

Method used

A magnetic bead-based sample preparation method was adopted, in which carboxyl magnetic beads were modified with N-hydroxysuccinimide and polyethyleneimine (PEI) to bind to arachidonic acid isomers in serum samples. Efficient separation and purification were achieved by covalent coupling and magnetic adsorption techniques, and analysis was performed by liquid chromatography-mass spectrometry.

Benefits of technology

It significantly improves the chromatographic separation resolution of isomeric metabolites, increases detection sensitivity by 10-100 times, shortens single sample processing time to within 30 minutes, and achieves a reaction conversion rate of 98%, laying the foundation for subsequent mass spectrometry-coupled detection.

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Abstract

The invention belongs to the technical field of detection, and particularly relates to a preparation method of a test sample for detecting arachidonic acid-like isomers based on magnetic beads, a test sample solution adopts a chemical derivatization reagent system, and covalent coupling magnetic beads are successfully used for nucleic acid and protein separation through surface modification. By accurately designing molecular recognition sites, the chromatographic separation resolution (delta R is larger than or equal to 1.5) of isomer metabolites can be remarkably improved, meanwhile, by means of an isotope double-labeling strategy, the detection sensitivity is improved by 10-100 times, the treatment time of a single sample is shortened to be within 30 minutes, and more importantly, by means of construction of a solid-phase reaction system, the detection sensitivity is greatly improved. Controllable optimization of derivatization reaction efficiency is achieved (the reaction conversion rate is larger than 98%), and a key technical foundation is laid for follow-up development of a full-automatic mass spectrum combined detection technology. The magnetic bead mediated solid phase derivatization strategy is expected to renovate the analysis normal form of small molecule metabolites, and promotes the metabonomics research to develop towards high flux and automation.
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