Hetero-Diels-Alder Cycloaddition Reaction of Vinyl Ethers Enables Selective Fluorescent Labeling of Plasmalogens in Human Plasma Lipids

Plasmalogens (Pls) are vinyl ether-containing glycerophospholipids of broad biological interest. Their abnormal levels are associated with neurological disorders and cardiovascular diseases. The intricacy of analyzing Pls in lipid samples arises from the wide variety of other coexisting lipid species, which underscores the urgent need for a Pls-specific labeling reaction. To address this challenge, we report an efficient hetero-Diels-Alder cycloaddition reaction between nonterminal vinyl ethers of Pls and o-quinolinone quinone methide probes under mild conditions. On the basis of this mechanism, a selective fluorescent labeling method for Pls is developed. The application of this method permits the exclusive derivatization of Pls over other human plasma lipids. The process also imparts labeled Pls with distinct fluorescence emission and chromatographic retention properties. By integrating this method with high-performance liquid chromatography, we are able to identify individual chromatographic signatures of Pls from 10 different human plasma samples. This Pls signature analytical technique, empowered by the Pls-specific labeling reaction, is cost-effective and simple in terms of instrumentation, suggesting its promising potential for the early screening and diagnosis of diseases linked to Pls abnormalities.

Automated summary

In this study, a highly efficient fluorescent labeling method based on hetero-Diels-Alder cycloaddition reaction is developed for selective labeling and identification of Plasmalogens (Pls) from complex lipid samples. By combining this method with high-performance liquid chromatography, individual chromatographic signatures of Pls from human plasma samples can be accurately identified. This technique is cost-effective and simple in terms of instrumentation, suggesting its promising potential for early screening and diagnosis of diseases associated with Pls abnormalities.