Fast sampling method for mammalian cell metabolic analyses using liquid chromatography-mass spectrometry

Area of Science:

• Metabolomics
• Cellular Biology
• Analytical Chemistry

Background:

• Metabolomics is crucial for biological research.
• Liquid chromatography-mass spectrometry (LC-MS) is a key technique for metabolic characterization.
• Efficient sample preparation is vital for accurate LC-MS data analysis.

Purpose of the Study:

• To present a robust and adaptable sample preparation workflow for mammalian cell metabolomics.
• To enable accurate qualitative and quantitative central metabolite characterization.
• To optimize sample handling for LC-MS analysis.

Main Methods:

• Utilizing mammalian cells grown on cover glasses for easy transfer.
• Implementing a rapid washing step in flowing water to reduce contamination and matrix effects.
• Employing a cold methanol-acetonitrile mixture with formic acid for rapid metabolic quenching, followed by freeze-drying and metabolite extraction.

Main Results:

• The protocol allows for fast cell sampling (2 seconds) and quenching.
• The entire workflow, including metabolite extraction and LC-MS, takes approximately 1.5 hours per sample.
• The method is suitable for adherent mammalian cell cultures.

Conclusions:

• This protocol provides an efficient and reliable method for mammalian cell sample preparation in metabolomics.
• It minimizes sample perturbation and enhances data accuracy for LC-MS analyses.
• The adaptability of the workflow supports various metabolic studies, including labeling experiments.