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Comparison of fully‐wettable RPLC stationary phases for LC‐MS‐based cellular metabolomics

Reversed‐phase LC combined with high‐resolution mass spectrometry (HRMS) is one of the most popular methods for cellular metabolomics studies. Due to the difficulties in analysing a wide range of polarities encountered in the metabolome, 100%‐wettable reversed‐phase materials are frequently used to maximise metabolome coverage within a single analysis. Packed with silica‐based sub‐3 μm diameter particles, these columns allow high separation efficiency and offer a reasonable compromise for metabolome coverage within a single analysis. While direct performance comparison can be made using classical chromatographic characterisation approaches, a comprehensive assessment of the column's performance for cellular metabolomics requires use of a full LC‐HRMS workflow in order to reflect realistic study conditions used for cellular metabolomics. In this study, a comparison of several reversed‐phase LC columns for metabolome analysis using such a dedicated workflow is presented. All columns were tested under the same analytical conditions on an LC‐Time‐of‐Flight‐MS (LC‐TOFMS) platform using a variety of authentic metabolite standards and biotechnologically‐relevant yeast cell extracts. Data on total workflow performance including retention behaviour, peak capacity, coverage, and molecular feature extraction repeatability from these columns are presented with consideration for both non‐targeted screening and differential metabolomics workflows using authentic standards and Pichia pastoris cell extract samples.

This article is protected by copyright. All rights reserved

Authors:   Le Si‐Hung, Tim J. Causon, Stephan Hann
Year:   2017
Pages:   n/a
DOI:   10.1002/elps.201700157
Publication date:   10-Jul-2017
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  • Single
  • Pichia pastoris
  • particles
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