Premium
Using i RT , a normalized retention time for more targeted measurement of peptides
Author(s) -
Escher Claudia,
Reiter Lukas,
MacLean Brendan,
Ossola Reto,
Herzog Franz,
Chilton John,
MacCoss Michael J.,
Rinner Oliver
Publication year - 2012
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201100463
Subject(s) - mass spectrometry , peptide , selected reaction monitoring , computer science , retention time , multiplexing , dimensionless quantity , software , chromatography , chemistry , biological system , computational biology , tandem mass spectrometry , biology , physics , biochemistry , mechanics , programming language , telecommunications
Multiple reaction monitoring ( MRM ) has recently become the method of choice for targeted quantitative measurement of proteins using mass spectrometry. The method, however, is limited in the number of peptides that can be measured in one run. This number can be markedly increased by scheduling the acquisition if the accurate retention time ( RT ) of each peptide is known. Here we present i RT , an empirically derived dimensionless peptide‐specific value that allows for highly accurate RT prediction. The i RT of a peptide is a fixed number relative to a standard set of reference i RT ‐peptides that can be transferred across laboratories and chromatographic systems. We show that i RT facilitates the setup of multiplexed experiments with acquisition windows more than four times smaller compared to in silico RT predictions resulting in improved quantification accuracy. i RT s can be determined by any laboratory and shared transparently. The i RT concept has been implemented in S kyline, the most widely used software for MRM experiments.