Targeted metabolomics profiling and quantitation solutions

Targeted profiling and quantitation solutions

Identified biomarkers from either untargeted metabolomics experiments, or hypothesis-driven from the literature, can be verified or validated where usually a limited number of compounds are quantified in a large set of samples.

For highest selectivity and confirmation, HRAM Orbitrap mass spectrometry is the technology of choice for verification of putative biomarkers. Moving through to high-throughput quantitation, SRM analysis on the TSQ triple quadrupole mass spectrometers is the solution of choice.

Popular metabolomics products

Standardized and quantitative HRAM LC-MS metabolomics workflow

Targeted metabolomics offers researchers the possibility to quantify selected metabolites. The AbsoluteIDQ® p400 HR Kit is the first standardized solution for broad metabolic and lipid profiling developed for Thermo Scientific high-resolution, accurate mass (HRAM) Orbitrap MS. It covers hundreds of metabolites while providing quantification, reproducibility, and high throughput. The kit offers the opportunity to analyze various biological matrices from multiple species.

Sample preparation

Biocrates AbsoluteIDQ p400 HR Kit

  • Ready-to-use kit developed for Q Exactive Focus LC-MS, Q Exactive LC-MS, Q Exactive Plus and Q Exactive HF LC-MS
  • Delivering a standardized, quality-controlled and reproducible quantitative assay
  • Provides excellent inter-laboratory reproducibility
  • Allows quantitation of key metabolites relevant to pathophysiological processes and disease specific markers

First steps

Samples are registered in the MetIDQ™ software (integral part of the kit). The test mix is run to ensure suitable system conditions.

Loading

Samples and reagents are pipetted onto the kit plate

Derivatization

Amino acids and biogenic amines are derivatized to help ensure a reliable analysis.

Extraction

Metabolites are extracted with organic buffer and diluted in two steps for subsequent mass spectrometric analysis.

LC-MS and FIA-MS Analysis

Q Exactive Orbitrap LC-MS Systems

The HRAM unique capabilities of Orbitrap technology complements the analysis by providing:

  • Resolution and accurate quantitation of isobaric lipid species
  • Increased selectivity and specificity for lipid analysis using flow injection analysis (FIA)
  • Accurate quantitation with less interferences
  • Partnered with Vanquish UHPLC for high resolution chromatographic separation and excellent retention time reproducibility

Data analysis

Biocrates MetIDQ™ Software

MetIDQ™ highly automates the whole FIA-MS and LC-MS process from project and sample administration to plate layout and work list generation, technical validation and statistical analysis to standardize metabolomics project data processing. MetIDQ is an integral part of all AbsoluteIDQ® Kits.

  • Import LC-MS results files from Thermo Scientific Xcalibur software
  • Direct and improved export to SPSS, XLSX, CSV, and XML
  • Improved project and export format (DMP) for large projects