Instruments

Mass Spectrometry Equipment Available in the Arizona Metabolomics Laboratory, Center for Metabolic and Vascular Biology (Mayo, Scottsdale Campus), Arizona State University

Agilent 1290 UPLC-6490 QQQ MS We have an Agilent 1290 UPLC-6490 QQQ MS system for highly multiplexed targeted metabolomics and flux analysis. This instrument is equipped with an UPLC system and HILIC or reverse phase chromatography columns for either aqueous or lipid metabolomics analysis. The molecules are ionized using an ESI source and MS analysis is performed in multiple-reaction-monitoring (MRM) mode. This instrument is used for detecting a variety of metabolites including amino acids, kynurenine pathway metabolites, acyl-carnitines, cardiolipins, and acetyl-CoAs. In addition, we have strong method development capabilities that are highly useful for various studies, such as globally optimized targeted mass spectrometry (GOT-MS), an approach we have introduced that maximizes the metabolite coverage using a targeted LC-MS/MS system. The instrument has high sensitivity and high scanning speed, and is installed with MassHunter Workstation software for MS data collection and data analysis.

Agilent 7820 GC-5977 MS This system is used to analyze volatile species as well as semi-volatile or non-volatize metabolites after chemical derivatization. Importantly, GC-MS is a classic method to collect data for flux analysis. EI spectra are quite reproducible, and thus the GC-MS system can easily identify over 150 metabolites by comparing the spectra of samples against those from the commercial NIST and Fiehn libraries available in the laboratory. The Agilent 5977 is a quadruple MS detector and it has excellent performance for quantitative purposes. Derivatization using MSTFA, MTBSTFA, and other silylation methods will strictly follow the corresponding standard operating procedures developed and validated by personnel in Gu’s lab.

Thermo UPLC Electron Orbitrap Elite Velos Pro MS We have a Thermo Scientific Electron Orbitrap Elite Velos Pro UPLC-MS system with strong metabolomics global profiling capabilities. This Orbitrap system can be used for untargeted global metabolite profiling, global flux analysis, and metabolite identification. The fast scanning ensures compatibility with narrow chromatographic peaks from UPLC separations. The system has enhanced ion optics that increase sensitivity and reliability, a greater dynamic range for better precursor detection, and the power of MSⁿ identification. This system has good resolution (>240k) and excellent mass accuracy (<2ppm) for many metabolites. The instrument has multiple collision modes (CID and HCD) that allow improved unknown identification and structural characterization.