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|Title:||Retention Order Reversal of Phosphorylated and Unphosphorylated Peptides in Reversed-Phase LC/MS|
KROKHIN, Oleg V.
|Author's alias:||小形, 公亮|
retention order reversal
linear solvation strength theory
|Publisher:||Japan Society for Analytical Chemistry|
|Journal title:||Analytical sciences : the international journal of the Japan Society for Analytical Chemistry|
|Abstract:||Protein phosphorylation is one of the most ubiquitous post-translational modifications in humans, and trypsin-digested phosphorylated peptides have been analyzed by reversed phase LC/MS using C18-silica columns under acidic conditions to profile human phosphoproteomes. Here, we report that phosphopeptides generally exhibit stronger retention than their unphosphorylated counterparts when C18-silica columns are used with acetic acid or formic acid as an ion-pairing reagent, whereas the retention order is reversed when less hydrophobic stationary phases such as C4-silica columns are employed. Similarly the retention reversal is observed when more hydrophobic ion-pairing reagents such as trifluoroacetic acid are used with C18-silica columns. These phenomena could be explained by the smaller S-values of phosphopeptides in linear solvation strength theory, based on the reduced net charge caused by intramolecular interaction between phosphate and basic groups.|
|Rights:||© 2018 by The Japan Society for Analytical Chemistry|
|Appears in Collections:||Journal Articles|
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