title: Extraction parameters for metabolomics from cultured cells.
abstract: 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.
authors.affiliation: Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Ser
authors.affiliation: Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Liu
authors.affiliation: Field of Genetics, Genomics, and Development, Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Ngoc Nu
authors.initials: NN
authors.lastname: Tang
authors.affiliation: Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA; Field of Genetics, Genomics, and Development, Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA. Electronic address: locasale@cornell.edu.
authors.forename: Jason W
authors.initials: JW
authors.lastname: Locasale
mesh_terms: D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D006801:Humans; D013058:Mass Spectrometry; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*
keywords: Extraction; Metabolomics