GC-MS/MS survey of collision-induced dissociation of tert-butyldimethylsilyl-derivatized amino acids and its application to 13C-metabolic flux analysis of Escherichia coli central metabolism

• Published in: Analytical and bioanalytical chemistry Vol. 408; no. 22; pp. 6133 - 6140
• Main Authors: Okahashi, Nobuyuki, Kawana, Shuichi, Iida, Junko, Shimizu, Hiroshi, Matsuda, Fumio
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 2016
• Subjects: Analytical Chemistry; Biochemistry; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Food Science; Laboratory Medicine; Monitoring/Environmental Analysis; Research Paper; Amino acids; C-metabolic flux analysis; GC-MS/MS; Fragmentation
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-016-9724-4

Stable isotope labeling experiments using mass spectrometry have been employed to investigate carbon flow levels (metabolic flux) in mammalian, plant, and microbial cells. To achieve a more precise 13 C-metabolic flux analysis ( 13 C-MFA), novel fragmentations of tert -butyldimethylsilyl (TBDMS)-amino acids were investigated by gas chromatography-tandem mass spectrometry (GC-MS/MS). The product ion scan analyses of 15 TBDMS-amino acids revealed 24 novel fragment ions. The amino acid-derived carbons included in the five fragment ions were identified by the analyses of 13 C-labeled authentic standards. The identification of the fragment ion at m/z 170 indicated that the isotopic abundance of S -methyl carbon in methionine could be determined from the cleavage of C5 in the precursor of [M–159] + ( m/z 218). It was also confirmed that the precision of 13 C-MFA in Escherichia coli central carbon metabolism could be improved by introducing 13 C-labeling data derived from novel fragmentations. Graphical Abstract Novel collision-induced dissociation fragmentations of tert -butyldimethylsilyl amino acids were investigated and identified by GC-MS/MS