Polar metabolomics profiling and fingerprinting methodology

Scope of the method

The Method relates to
  • Animal health
  • Environment
  • Human health
The Method is situated in
  • Basic Research
  • Translational - Applied Research
Type of method
  • In chemico
Metabolomics
This method makes use of
  • Animal derived cells / tissues / organs

Description

Method keywords
  • metabolomics
  • mass spectrometry
  • Liquid chromatography
  • metabolic profiling
  • Metabolism
  • metabolic fingerprinting
  • biofluids
  • urine
  • feces
  • saliva
  • blood
  • cell culture
  • colon tissue
Scientific area keywords
  • analytical chemistry
  • metabolic disorders
  • inflammation
  • biofluids
  • cancer research
  • food allergy
Method description

Our polar metabolomics profiling and fingerprinting methodology applies ultra-high performance liquid chromatography coupled to hybrid quadrupole-Orbitrap high resolution mass spectrometry. Both the instrumental method, as well as generic extraction protocols for colon tissue, cell cultures, urine, feces, plasma and saliva have been extensively validated in both a targeted as well as an untargeted fashion. The metabolomics workflow consists of a sample preparation, followed by the UPHLC-HRMS analysis, after which multivariate statistical analysis will be performed to identify potential biomarker candidates or altered pathways, associated with a specific metabolic state.

Lab equipment
  • HPLC ;
  • HR-Orbitrap-MS.
Method status
  • Internally validated
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages

The metabolome is considered as the endpoint of metabolism and is therefore influenced by amongst others the genes, the diet, the environment and the residing microbiome. As such, the measurement of the metabolome provides the most holistic image of the phenotype of a patient. Additionally, it provides both a qualitative as well as a quantitative functional read-out. Therefore, it can be considered the method of choice for hypothesis testing and hypothesis generation.

Challenges
  • Multi-step procedure => Long analysis time, extensive sample preparation ;
  • Big data handling.
Modifications

The method can be adapted to other matrices or other animal species when necessary.

References, associated documents and other information

References

Vanden Bussche et al (2015)

Analytical Chemistry, 87, 10927-10934 De Paepe et al (2018)

Analytica Chimica Acta, 1033, 108-118 Rombouts et al (2019)

Analytica Chimica Acta, 1066, 79-92 Wijnant et al (2019) submitted

De Spiegeleer et al (2019) submitted

Associated documents
De Paepe et al, 2018.pdf
Rombouts et al, 2019.pdf
Vandenbussche et al, 2015.pdf
Links
Rombouts et al, 2019
De Paepe et al, 2018
Vanden Bussche et al, 2015

Contact person

Lynn Vanhaecke

Organisations

Ghent University
Faculty of Veterinary Medicine, Department of Veterinary Public Health and Food Safety
Belgium
Flemish Region