Classical Untargeted Metabolomics by UPLC-FT-MS

---

Description

This service uses ultrahigh-performance liquid chromatography (UPLC), in the separation modes of reversed-phase, hydrophilic interaction and/or ion-pairing LC, coupled to high-resolution (60,000 or 12,0000 FWHM) Orbitrap-type Fourier Transform mass spectrometry (FTMS) in both (+) and (-) ion detection modes to relatively quantify up to 15,000 metabolite features in typical biological samples with >4000 assigned metabolites.

QC samples are injected multiple times throughout the analysis to monitor instrument stability (platform variability, retention time shift, mass shift, etc) and sample stability to ensure high-quality datasets. QC data is evaluated for each data set with multivariate statistics.

Identification of metabolite biomarkers following statistics and multivariate analysis (PCA, PLS-DA, and OPLS) is performed by UPLC-MS/MS with collision-induced dissociation (CID) and high-energy C-trap dissociation (HCD) on one of two LTQ-Orbitrap mass spectrometers (Velos Pro, Fusion), with the aid of a library of standard substances of >1,200 metabolites in stock.

Platform Strengths

Relative and precise quantitation of hydrophilic to lipophilic metabolites and lipids by untargeted metabolomics analysis combined with database searching and putative metabolite IDs based on accurate masses.

Quantitative

Discovery (relative quantification)

Metabolite Identification (Tier 1, Tier 2, Tier 3)

We primarily provide identification for metabolites that have been identified to be statistically relevant.

Tier 3 is provided as optional (upon request) by using accurately measured masses (errors ≤4 ppm) to query metabolome databases for preliminary metabolite assignments with putative identities.

Tier 1 and Tier 2 is provided to identify and structurally confirm statistics relevant metabolite biomarkers by combining metabolome/lipidome database searching, LC-MS and LC-MS/MS spectral matching and interpretation, with the aid of in-stock authentic compounds of >1,200 metabolites.

Additional fees may apply in cases where additional LC-MS and LC-MS/MS runs are needed in follow-on experiments or when new standards need to be acquired to confirm the idenity of novel metabolites not currently in our library.

Example Data

Serum/plasma: Tier 3-5229 metabolites and lipids

Statistical Analysis

T-test statistics and PCA or PLS-DA are included. Additional statistics and pathway analysis is available upon request.

Platform Available

UPLC-Orbitrap-MS

---