Lipidomics 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
Lipidomics
This method makes use of
  • Animal derived cells / tissues / organs

Description

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

Our lipidomics profiling and fingerprinting methodology enables the analysis of all lipids within a biological matrix. These lipids can then be compared between an individual or condition with a specific metabolic state and healthy controls to find biomarkers or pathways that can be related to diseases. It 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, blood, urine and feces have been extensively validated in both a targeted as well as an untargeted fashion. The lipidomics 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
  • UHPLC ;
  • HR-Orbitrap-MS.
Method status
  • History of use
  • Internally validated
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages

Lipids are involved in a plethora of biological processes, including energy homeostasis, immune response, membrane architecture, enzyme activity, inflammation, cellular signaling and transduction of cellular events. Evidently, a dysregulated lipid metabolism has been implicated in a variety of pathological conditions. Therefore, the measurement of this lipidome, which is at the end of the genome-transcriptome-proteome cascade, will provide 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
  • Van Meulebroek et al (2017)
  • Analytical Chemistry Rombouts et al (2019) 
  • Analytica Chimica Acta De Spiegeleer et al (2019) submitted
Associated documents
Van Meulebroek et al. 2017.pdf
Rombouts et al, 2019.pdf
Links
Van Meulebroek et al, 2017
Rombouts et al, 2019

Contact person

Lynn Vanhaecke

Organisations

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