title: Optimized precursor ion selection for labile ions in a linear ion trap mass spectrometer and its impact on quantification using selected reaction monitoring.
abstract: The fragmentation of fragile ions during the application of an isolation waveform for precursor ion selection and the resulting loss of isolated ion intensity is well-known in ion trap mass spectrometry (ITMS). To obtain adequate ion intensity in the selected reaction monitoring (SRM) of fragile precursor ions, a wider ion isolation width is required. However, the increased isolation width significantly diminishes the selectivity of the channels chosen for SRM, which is a serious problem for samples with complex matrices. The sensitive and selective quantification of many lipid molecules, including ceramides from real biological samples, using a linear ion trap mass spectrometer is also hindered by the same problem because of the ease of water loss from protonated ceramide ions. In this study, a method for the reliable quantification of ceramides using SRM with near unity precursor ion isolation has been developed for ITMS by utilizing alternative precursor ions generated by in-source dissociation. The selected precursor ions allow the isolation of ions with unit mass width and the selective analysis of ceramides using SRM with negligible loss of sensitivity. The quantification of C18:0-, C24:0- and C24:1-ceramides using the present method shows excellent linearity over the concentration ranges from 6 to 100, 25 to 1000 and 25 to 1000 nM, respectively. The limits of detection of C18:0-, C24:0- and C24:1-ceramides were 0.25, 0.25 and 5 fmol, respectively. The developed method was successfully applied to quantify ceramides in fetal bovine serum.
authors.affiliation: College of Pharmacy, Chungbuk National University, 52 Naesudong-ro, Heungdeok-gu, Cheongju, Chungbuk, 361-763, Korea; Korea Research Institute of Standards and Science, 267 Gajeong-ro, Yuseong-gu, Daejeon, 305-340, Korea.
authors.forename: Hyun-Seok
authors.initials: HS
authors.lastname: Lee
authors.forename: Kyong-Oh
authors.initials: KO
authors.lastname: Shin
authors.forename: Sung-Chan
authors.initials: SC
authors.lastname: Jo
authors.forename: Yong-Moon
authors.initials: YM
authors.lastname: Lee
authors.forename: Yong-Hyeon
authors.initials: YH
authors.lastname: Yim
mesh_terms: D000818:Animals; D002417:Cattle; D002518:Ceramides / Q000032:analysis / Q000097:blood*; D007477:Ions / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002518:Ceramides; D007477:Ions
keywords: ceramides; fragile ion; in-source dissociation; ion trap mass spectrometer; selected reaction monitoring