Abstract: Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has been one of the most popular approaches in proteome analysis. As an independent parameter to mass spectrometry information, peptide retention time has been utilized to facilitate protein identification and quantification. In the field of peptide identification, the prediction of the retention time combined with routine tandem mass spectrometry database searching methods could help improve the confidence of identification. The sensitivity of identification could also be improved by matching peaks with both the accurate mass and retention time in multiple aligned LC-MS runs. Meanwhile, because small changes of liquid chromatography conditions lead to variability in retention times unavoidably, retention time alignment is crucial to label-free quantification. Additionally, post-translational modifications (PTM) could be identified by combining retention time shifts and mass deviation information.

Key words: label-free quantification, peptide identification, post-translational modification (PTM), proteomics , retention time alignment, retention time prediction, liquid chromatography-tandem mass spectrometry (LC-MS/MS)