One‐Pot Construction of NHS‐Activated Magnetic Particles for Chemoselective Capture of Carboxyl Metabolites

Chemoselective probes immobilize on magnetic materials show great promise in simplifying sample handling and enhancing detection sensitivity. However, their complicated preparation and associated expense limit broader application. In this study, novel magnetic particles with abundant N-hydroxysuccinimide (NHS) esters on the surface are conveniently synthesized using a one-pot method without carbodiimide activation carboxylate molecules. Subsequently, multifunctional probes are synthesized by immobilizing high-density chemical probes on the surface of the magnetic materials through a postsynthetic modification strategy. This versatile probe facilitates the rapid capture of carboxylated compounds from complex matrices, with the labeled metabolites release from the magnetic materials subsequently analyzed using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS). The advantages of this innovative chemical biological tool include the simplicity and low cost of the synthesis, as well as the capability to analyze polar and volatile carboxylated metabolites via LC-MS. This new strategy is successfully applied to analyze short-chain fatty acids (SCFAs) in rat cecal contents, demonstrating the reliability of the analytical method. This study presents a cost-effective and easy-to-implement approach for preparing NHS-activated magnetic particles and offers a versatile probe with chemoselective extraction and labeling capabilities, providing a practical tool for analyzing SCFAs in the gut.