Genomes to Natural Products Network (GNPN):
Stanford Genome Technology Center


Yeast Organic Extraction for LC-MS

Method Overview

This protocol outlines a quick and simple procedure for extracting all organic compounds from a yeast culture for subsequent analysis via Liquid chromatography–mass spectrometry (LC-MS). It is suitable for quickly assessing the presence of products from small sample preps.

Materials:

Organic solvents: Ethyl Acetate, Acetone, Acetic Acid and Methanol

Protocol

  1. Grow yeast in selective medium overnight.

  2. Inoculate a 30mL YPD culture to a final OD600 of ~0.2.

  3. Grow at 30°C, shaking at 200 rpm.

  4. At day 3 and/or 4, harvest 2mL of culture in 15 mL Falcon tubes. Spin down.

  5. Move the supernatant to a new tube and add 2mL of ethyl acetate. Mix by vortexing and incubate at RT overnight.

  6. For the pellet, add 1mL of acetone and 1mL of ethyl acetate. Mix by vortexing and incubate at RT overnight.

  7. For unknown compounds, consider adding vol/vol 1% of acetic acid.

  8. Take 0.5 mL from each extraction, avoiding pellet or particulate matter. Speed vacuum at RT until completely dry.
  9. Redissolve in 200µL of methanol. Load 5-8µL on LC-MS.


Inquiries can be addressed to Maureen Hillenmeyer (maureenh at stanford.edu) and Angela Chu (amchu at stanford.edu)
Stanford Genome Technology Center