One microliter of reaction supernatant was mixed with an equal volume of 20 mg mL−1 2,5-dihydroxybenzoic acid (DHB) in 50% acetonitrile, 0.1% TFA on a SCOUT-MTP 384 target plate (Bruker). The spotted samples were then dried in a vacuum desiccator before being analyzed by mass spectrometry on an Ultraflex III matrix-assisted laser desorption ionization-time of flight/time of flight (MALDI/TOF-TOF) instrument (Bruker)54 (link).
Sample permethylation was carried out according to Ciucanu and Kerek55 (link). Spotted samples were analyzed by MS using 2,5-DHB matrix with 0.1% TFA on an AB-Sciex 4700 (for MALDI-TOF) and Ultraflex III MALDI/TOF-TOF instrument (Bruker). Data were collected using a 2-kHz smartbeam-II laser and acquired on reflector mode (mass range 300–3000 Da) for MS analysis and on LIFT-CID for MS/MS analysis using argon as collision gas. FlexControl and FlexAnalysis softwares were used for data acquisition and analysis. On average, about 10,000 shots were used to obtain high-enough resolution. MS/MS fragmentation patterns were named according to Domon and Costello56 (link).
Sample permethylation was carried out according to Ciucanu and Kerek55 (link). Spotted samples were analyzed by MS using 2,5-DHB matrix with 0.1% TFA on an AB-Sciex 4700 (for MALDI-TOF) and Ultraflex III MALDI/TOF-TOF instrument (Bruker). Data were collected using a 2-kHz smartbeam-II laser and acquired on reflector mode (mass range 300–3000 Da) for MS analysis and on LIFT-CID for MS/MS analysis using argon as collision gas. FlexControl and FlexAnalysis softwares were used for data acquisition and analysis. On average, about 10,000 shots were used to obtain high-enough resolution. MS/MS fragmentation patterns were named according to Domon and Costello56 (link).
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