Chemoselective and Highly Sensitive Quantification of Gut Microbiome and Human Metabolites

Abstract

The microbiome has af undamental impact on the human hostsp hysiology through the production of highly reactive compounds that can lead to disease development. One class of such compounds are carbonyl-containing metabolites, which are involved in diverse biochemical processes.M ass spectrometry is the method of choice for analysis of metabolites but carbonyls are analytically challenging. Herein, we have developed an ew chemical biology tool using chemoselective modification to overcome analytical limitations.T wo isotopic probes allowf or the simultaneous and semi-quantitative analysis at the femtomole level as well as qualitative analysis at attomole quantities that allows for detection of more than 200 metabolites in human fecal, urine and plasma samples. This comprehensive mass spectrometric analysis enhances the scope of metabolomics-driven biomarker discovery.Weanticipate that our chemical biology tool will be of general use in metabolomics analysis to obtain ab etter understanding of microbial interactions with the human host and disease development.

Coral-like Magnetic Particles for Chemoselective Extraction of Anionic Metabolites

“…An ideal chemical derivatization method should provide better chromatographic separation and improve the ionization efficiency of metabolites, thereby significantly enhancing the detection sensitivity of liquid chromatography–MS (LC–MS) methods and facilitating targeted metabolite detection and identification. − However, the methods described so far have to contend with interference from the sample background and excess derivatization reagents. Selective discovery of substantial metabolites at low concentrations in biological samples is critical for better understanding the entire metabolome. , Few advanced analytical methods, such as surface-enhanced Raman spectroscopy , and microdialysis probes, , are available to study low-abundance or difficult-to-detect metabolites. In contrast, other “omics” research fields, such as proteomics and genomics, benefit from a variety of chemical biology tools and chemical approaches. , …”

“…Selective discovery of substantial metabolites at low concentrations in biological samples is critical for better understanding the entire metabolome. , Few advanced analytical methods, such as surface-enhanced Raman spectroscopy , and microdialysis probes, , are available to study low-abundance or difficult-to-detect metabolites. In contrast, other “omics” research fields, such as proteomics and genomics, benefit from a variety of chemical biology tools and chemical approaches. , …”

“…In 2018, Globisch et al reported a unique chemoselective probe immobilized onto MNPs, which can specifically capture metabolites from biological samples and release them under mild conditions . The isolation from the sample matrix could significantly reduce ion suppression, enhance the detection sensitivity, and improve detection limits of metabolites at low concentrations …”

“…In contrast, other "omics" research fields, such as proteomics and genomics, benefit from a variety of chemical biology tools and chemical approaches. 6,12 Because of their unique properties, magnetic nanoparticles (MNPs) play an important role in the design of innovative methods and tools, 17,18 especially in bioanalysis and medical sciences. 19,20 MNPs have shown huge potential to solve fundamental biological and medical problems in the fields of diagnostics and therapy.…”

“…6 The isolation from the sample matrix could significantly reduce ion suppression, enhance the detection sensitivity, and improve detection limits of metabolites at low concentrations. 12 Central carbon metabolism (CCM), composed of glycolysis, the pentose phosphate pathway (PPP), and the tricarboxylic acid (TCA) cycle, plays a key role in substrate degradation, energy and cofactor regeneration, and the supply of biosynthetic precursors (DNA, RNA, etc.). 29 Almost all the components involved in the central carbon and energy metabolism are negatively charged, such as phosphorylated saccharides, phosphorylated carboxylic acids, carboxylates, phosphates, and nucleotides.…”