RESUMEN
The microbiome has a fundamental impact on the human host's physiology 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. Mass spectrometry is the method of choice for analysis of metabolites but carbonyls are analytically challenging. Herein, we have developed a new chemical biology tool using chemoselective modification to overcome analytical limitations. Two isotopic probes allow for 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. We anticipate that our chemical biology tool will be of general use in metabolomics analysis to obtain a better understanding of microbial interactions with the human host and disease development.
Asunto(s)
Acetaldehído/análisis , Acetona/análisis , Aldehídos/análisis , Butanonas/análisis , Dihidroxiacetona/análisis , Metabolómica/métodos , Acetaldehído/sangre , Acetaldehído/química , Acetaldehído/orina , Acetamidas/química , Acetona/sangre , Acetona/química , Acetona/orina , Aldehídos/sangre , Aldehídos/química , Aldehídos/orina , Butanonas/sangre , Butanonas/química , Butanonas/orina , Carbono/química , Isótopos de Carbono/química , Dihidroxiacetona/sangre , Dihidroxiacetona/química , Dihidroxiacetona/orina , Heces/química , Microbioma Gastrointestinal , Humanos , Indicadores y Reactivos/química , Límite de Detección , Orina/químicaRESUMEN
Desorption electrospray ionization mass spectrometry (DESI-MS) and nuclear magnetic resonance (NMR) spectroscopy are used to provide data on urine examined without sample preparation to allow differentiation between diseased (lung cancer) and healthy mice. Principal component analysis (PCA) is used to shortlist compounds with potential for biomarker screening which are responsible for significant differences between control urine samples and samples from diseased animals. Similar PCA score plots have been achieved by DESI-MS and NMR, using a subset of common detected metabolites. The common compounds detected by DESI and NMR have the same changes in sign of their concentrations thereby indicating the usefulness of corroborative analytical methods. The effects of different solvents and surfaces on the DESI mass spectra are also evaluated and optimized. Over 80 different metabolites were successfully identified by DESI-MS and tandem mass spectrometry experiments, with no prior sample preparation.
