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Capillary LC coupled with high-mass measurement accuracy mass spectrometry for metabolic profiling

Reputable Mentor II
Reputable Mentor II
Ding J, Sorensen CM, Zhang Q, Jiang H, Jaitly N, Livesay EA, Shen Y, Smith RD, Metz TO.
Anal Chem. 2007 Aug 15;79(16):6081-93.
We have developed an efficient and robust high-pressure capillary LC-MS method for the identification of large numbers of metabolites in biological samples using both positive and negative ESI modes. Initial efforts focused on optimizing the separation conditions for metabolite extracts using various LC stationary phases in conjunction with multiple mobile-phase systems, as applied to the separation of 45 metabolite standards. The optimal mobile and stationary phases of those tested were determined experimentally (in terms of peak shapes, theoretical plates, retention of small, polar compounds, etc.), and both linear and exponential gradients were applied in the study of metabolite extracts from the cyanobacterium Cyanothece sp. ATCC 51142. Finally, an automated dual-capillary LC system was constructed and evaluated for the effectiveness and reproducibility of the chromatographic separations using the above samples. When coupled with a commercial LTQ-orbitrap MS, approximately 900 features were reproducibly detected from Cyanothece sp. ATCC 51142 metabolite extracts. In addition, 12 compounds were tentatively identified, based on accurate mass, isotopic distribution, and MS/MS information.
Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
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‎10-15-2021 11:26 AM
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