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Universal retention standard for peptide separations using various modes of high-performance liquid chromatography
Ist Teil von
Journal of Chromatography A, 2019-03, Vol.1588, p.163-168
Ort / Verlag
Netherlands: Elsevier B.V
Erscheinungsjahr
2019
Link zum Volltext
Quelle
Elsevier ScienceDirect Journals Complete
Beschreibungen/Notizen
•Modeling peptide properties helps the development of peptide retention standard.•The standard can be used for the most popular peptide separations modes.•The list includes: RPHPLC, HILIC, strong anion and cation exchange, CZE.•Peptides have nearly identical mass and intense y ion to simplify MS assignments.
Peptide retention standards are widely used by chromatography specialists. They can be used for quality control of peptide separations (separation efficiency, selectivity, retention values) and for accurate concatenation of retention data from multiple acquisitions in proteomics. So far the repertoire of available retention standards is mostly limited to reversed-phase separations. We introduce a synthetic peptide mixture which can be used in conjunction with the most popular peptide separation techniques: reversed-phase (RPLC), strong-cation exchange (SCX), (strong-anion exchange) SAX and hydrophilic interaction liquid chromatography (HILIC). Target sequences were first designed in-silico using Sequence-Specific Retention Calculator models covering all major peptide separation mechanisms. Peptides were also designed while keeping in mind the simplicity of retention time assignment using MS detection: they all have nearly identical masses and identical intense y3 fragment ions. This contribution demonstrates the application of this mixture for characterization of eight HILIC as well as SAX, SCX and C18 columns.