Metabolomic profiling

An integrated platform for comparing metabolomic alterations between groups

Metabolome refers to the entire collection of small molecule metabolites within an organism. Profiling of metabolome is an important tool in system biology, biomarker discovery and also can be used to monitor the efficacy of drug intervention. We have developed an integrated platform for comparing metabolomic alterations between groups. The metabolites are extracted and separated on a reverse phase UPLC column followed by analysis on a Q-TOF mass spectrometer. The ion features are extracted, retention time aligned and normalized, then subjected to multivariate analysis. The metabolites with significant changes are searched against metabolomic databases such as Metlin and KEGG with accurate masses and isotopic pattern information, which returns putative metabolite identifications. To further evaluate the correctness of the resultant metabolite identifications from database search, MS/MS spectra are collected on these metabolites and manually compared with the chemical structure of the putative ids with the aid of fragmentation data analysis tool and empirical knowledge of gas-phase ion chemistry. For the definitive identification, the MS/MS spectra and retention times of the experimental data are compared with metabolite standards. Once the metabolites are identified, they can be quantitated by targeted analysis using selective reaction monitoring on our triple quadrupole instrument. The throughput for the targeted analysis is much higher so a large number of samples can be screened.