Extraction parameters for metabolomics from cultured cells.

Published

Journal Article

The successful extraction of metabolites is a critical step in metabolite profiling. By optimizing metabolite extraction, the range and quantitative capacity of metabolomics studies can be improved. We considered eight separate extraction protocols for the preparation of a metabolite extract from cultured mammalian cells. Parameters considered included temperature, pH, and cell washing before extraction. The effects on metabolite recovery were studied using a liquid chromatography high-resolution mass spectrometry (LC-HRMS) platform that measures metabolites of diverse chemical classes, including amino acids, lipids, and sugar derivatives. The temperature considered during the extraction or the presence of formic acid, a commonly used additive, was shown to have minimal effects on the measured ion intensities of metabolites. However, washing of samples before metabolite extraction, whether with water or phosphate-buffered saline, exhibited dramatic effects on measured intensities of both intracellular and extracellular metabolites. Together, these findings present a systematic assessment of extraction conditions for metabolite profiling.

Full Text

Duke Authors

Cited Authors

  • Ser, Z; Liu, X; Tang, NN; Locasale, JW

Published Date

  • April 15, 2015

Published In

Volume / Issue

  • 475 /

Start / End Page

  • 22 - 28

PubMed ID

  • 25613493

Pubmed Central ID

  • 25613493

Electronic International Standard Serial Number (EISSN)

  • 1096-0309

Digital Object Identifier (DOI)

  • 10.1016/j.ab.2015.01.003

Language

  • eng

Conference Location

  • United States