Assessment of capillary anion exchange ion chromatography tandem mass spectrometry for the quantitative profiling of the phosphometabolome and organic acids in biological extracts

Challenges and emergent solutions for LC‐MS/MS based untargeted metabolomics in diseases

“…The system also showed good stability and reproducibility, and the relative standard deviations of intensity and retention times for repeated analysis of six consecutive days were within 8%. Despite its advantages, IC‐MS has an issue of carry‐over and a fast‐gradient method with the injection of 50% methanol/water between analytical samples was needed to eliminate carry‐over and ensure optimal re‐equilibration of the column …”

Fast filtration sampling protocol for mammalian suspension cells tailored for phosphometabolome profiling by capillary ion chromatography – tandem mass spectrometry

“…During the development of the capIC-MS/MS method for phosphometabolome profiling it was observed that saline washed extracts were not compatible with capIC separation (i.e., large salt peak for the 10-15 first minutes, peak splitting, less stable retention times, and frequent replacement of expensive instrument parts such as column and suppressor), but this was successfully solved for adherently growing cells by including a fast DI-water rinse step that was proven tolerable by the cells [21]. Filtration has to be considered a tougher treatment for human cells as cells entrapped in filter materials are exposed to higher shear forces and faster osmotic changes than the washing of adherent cells.…”

“…The supernatant was carefully transferred to 3 kD molecular cutoff filters (#516-0228, VWR) and centrifuged at 14.000 rpm for 10 min. The peptide and protein free supernatant was then transferred into LC-MS vial inserts and run in a randomized order on a Dionex ICS-4000 capillary ion chromatograph coupled to a Waters Xevo TQ-S MS/MS according to Kvitvang et al [21]. The data was analyzed in MassLynx V4.1.…”

“…CapIC-MS/MS is the premium choice for quantitative phosphometabolome profiling [21,22]. Present protocols employ saline or phosphate buffered saline as final washing solutions prior to extraction, but these are not compatible with capIC-MS/MS analysis.…”

Advanced analytical methods and sample preparation for ion chromatography techniques

“…129 Applying a 0.32 Â 100 mm column packed with 3 mm octadecyl silica (ODS) microparticles coated with hexadecyltrimethylammonium bromide and mixed eluent containing 1 mM sodium salicylate with 5% acetonitrile, Takeuchi's group developed another CIC method for inorganic anions. [135][136][137][138][139] Using an injection valve to couple CIC and MS was found to be the best method for hyphenation, since it constituted a exible and dead-volume-free approach. The BSA molecule, immobilized on 5 mm ODS particles, was protonated at specic pH condition and affected the anionic analyte retention.…”