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Development of a solid phase extraction protocol coupled with liquid chromatography mass spectrometry to analyze central carbon metabolites in lake sediment microcosms
Author(s) -
Yang Song,
Synovec Robert E.,
Kalyuzhnaya Marina G.,
Lidstrom Mary E.
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100533
Subject(s) - chemistry , solid phase extraction , chromatography , mass spectrometry , extraction (chemistry) , microcosm , detection limit , sample preparation , ion exchange , environmental chemistry , ion , organic chemistry
Abstract An ion exchange solid phase extraction (SPE) strategy is developed for application with liquid chromatography‐tandem quadrupole mass spectrometry (LC‐MS/MS) for characterization of central carbon metabolites involved in methane assimilation and adjacent pathways in natural mixtures. For this purpose, short‐time microcosm samples were obtained from lake sediment known to consume methane. Three SPE procedures were developed for the recovery of 51 targeted metabolites from five compound classes (amino acids, carboxylic acids, sugar phosphates, nucleotides and acyl‐CoAs). The three SPE procedures employed were mixed mode (i) strong cation exchange, (ii) strong anion exchange and (iii) weak anion exchange. By spiking stable isotopic labeled standards, validation of the SPE procedures for the sediment extracts demonstrated that a 3 cm 3 , 60 mg SPE sorbent bed provided effective loading capacity for targeted metabolites with an analytical variation of 16% RSD. We readily analyzed 32 of the targeted 51 metabolites using LC‐MS/MS after sediment sample extraction, cleanup and pre‐concentration. The remaining 19 targeted metabolites were either at, or below, the limit of detection. The current approach provides a good workflow for absolute quantification of intermediates in C 1 ‐ carbon metabolism in natural microbial communities.