Metabolite Extraction from Saccharomyces cerevisiae for Liquid Chromatography-Mass Spectrometry

Area of Science:

• Metabolomics
• Cellular Biology
• Analytical Chemistry

Background:

• Accurate analysis of cellular small molecules requires effective extraction and separation from contaminants.
• Maintaining metabolic integrity during sample preparation is crucial for reliable metabolomic studies.

Purpose of the Study:

• To develop and describe a rapid, non-perturbing method for harvesting yeast cells.
• To enable simultaneous quenching of metabolism and extraction of small molecules.
• To ensure sample suitability for liquid chromatography-mass spectrometry (LC-MS) and long-term storage.

Main Methods:

• Rapid yeast cell harvesting via filtration from liquid culture.
• Simultaneous metabolic quenching and small molecule extraction using low-temperature organic solvent immersion.
• Preparation of samples for downstream liquid chromatography-mass spectrometry (LC-MS) analysis.

Main Results:

• The described method effectively harvests yeast cells while minimizing metabolic perturbation.
• Simultaneous quenching and extraction are achieved through immediate solvent immersion.
• Prepared samples demonstrate stability for over one year when stored at -80°C in solvent.

Conclusions:

• This filtration and solvent-immersion technique provides a robust method for yeast small molecule analysis.
• The approach is compatible with various liquid chromatography-mass spectrometry platforms.
• The method ensures high-quality, stable samples for metabolomic research.