Polar metabolites were derivatized and analysed by GC-MS (Agilent 7890B-5977A) and identification and abundance of individual metabolites was estimated as previously described 113 (link) . In brief, dried metabolite samples were washed with methanol (twice), and derivatized overnight at RT with methoxyamine (20 mg/mL in pyridine, Sigma) followed by addition of BSTFA + 1% TMCS (Sigma) for > 1 hr at RT. GC-MS was performed using splitless injection (injection temperature 270°C) onto a 30 m + 10 m × 0.25 mm DB-5MS+DG column (Agilent J&W), with helium carrier gas, in electron impact ionization mode. The oven temperature was initially 70°C (2 min), followed by a temperature increase to 295°C at 12.5°C/min and subsequently to 320°C at 25°C/min (held for 3 min). GAVIN software 114 (link) was used for metabolite identification and quantification by comparison to the retention times, mass spectra, and responses of known amounts of authentic standards.
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Vowinckel J., Hartl J., Marx H., Kerick M., Runggatscher K., Keller M.A., Mülleder M., Day J., Weber M., Rinnerthaler M., Yu J.S., Aulakh S., Lehmann A., Mattanovich D., Timmermann B., Zhang N., Dunn C.D., MacRae J.I., Breitenbach M, & Ralser M. (2021). The metabolic growth limitations of petite cells lacking the mitochondrial genome. Nature metabolism, 3(11), 1521-1535.
Other organizations :
University of Cambridge, BOKU University, Max Planck Institute for Molecular Genetics, Max Planck Unit for the Science of Pathogens, University of Salzburg, The Francis Crick Institute, Humboldt-Universität zu Berlin, Charité - Universitätsmedizin Berlin, Freie Universität Berlin, University of Helsinki
Use of GAVIN software for metabolite identification and quantification
controls
Comparison to retention times, mass spectra, and responses of known amounts of authentic standards
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