Metabolomics - Possible workflows

4 important questions on Metabolomics - Possible workflows

Explain how to go from sample to spectrum?

How do we measure them?
- Start with a biological sample
- Quenching = arresting metabolism
- Extraction procedure
- Chromatography (geeft pieken weer maar dit is niet uniek)
- Mass spectrometry (geeft massa/lading verhouding, info over de moleculaire structuur, kan hele lage concentraties meten)
- Translate result to metabolites

Why quenching; stopping the metabolism before analyzing?

Omdat je bij metabolomics de echte, directe toestand van de metabolieten op een bepaald tijdstip wilt meten — en metabolisme verandert per seconde.

Je stopt de enzymactiviteit, bevriest het metabolism.

How does sample preparation work? Wat zijn verschillende manieren van extractie? (3)

It is dependent on the matrix (soort materiaal, bloed, urine etc)
  • Liquid extraction = neerslaan van proteins
  • Liquid-liquid extraction = polair vs apolair
  • Solid phase extraction = verwijderen van zouten
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Why is sample preparation relevant?

- Selection of analytes of choice (physicalchemical properties)
- Sample clean-up
- Increase of sensitivity/ concentration

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