Assay: Untargeted LC-MS Metabolite Profiling
Our LC-MS Metabolite Profiling method (Untargeted Metabolomics) is carried out using UHPLC-Orbitrap-MS. A total of 2 UHPLC-MS runs are performed for each sample. Depending on sample type, two different columns (Reversed-phase and hydrophilic interaction liquid chromatography (HILIC)), mobile phase compositions, and ionization modes (positive and negative) may be used. Data provided is semi-quantitative in the form of peak area, adjusted for internal standards.
Metabolite spectra and associated retention times have been confirmed using chemical standards to generate an in-house spectral library (MSI Level 1). This is combined with NIST 20, MoNA, MassBank EU,and GNPS spectral libraries for metabolite identification. Total number of spectra from these combined libraries is over 1,000,000. Metabolites commonly identified include organic acids, alcohols, amines and polyamines, amino acids, acylcarnitines, nucleotides, bile acids, steroids, and phenolics.
Numerous surrogate and instrument internal standards are utilized. Study-specific pooled QC samples are included for intra- and inter-batch variability assessment, as well as data cleaning. BioIVT pooled plasma is used as a long-term reference QC sample for inter-study and inter-laboratory assessment of data.