title: The use of on-line capillary electrophoresis/electrospray ionization with detection via an ion trap storage/reflectron time-of-flight mass spectrometer for rapid mutation-site analysis of hemoglobin variants.
abstract: Capillary electrophoresis/electrospray ionization using an ion trap storage/reflectron time-of-flight mass spectrometer detector (CE/ESI-IT/reTOF) is used to provide a rapid and sensitive method for analyzing structural variants in the hemoglobin (Hb) beta-chain. The Hb alpha- and beta-chains are separated and the beta-chain is digested by trypsin. The digest is analyzed by CE/ESI-IT/reTOF where a comparison of the total ion electrophorograms and mass spectra of the mutant and normal hemoglobins (Hbs) can detect the presence of a mutation site. In addition, collision-induced dissociation in the vacuum interface-skimmer region can be used to pinpoint the identity of such a site. The unique capability of the CE/ESI-IT/reTOF system for accurately detecting fast separations with narrow peaks that may be under 1 s full width at half maximum is demonstrated. The speed of this system is essential for resolution of the large number of peaks that are separated in a short time duration using CE separations.
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor 48109-1055, USA.
authors.forename: M X
authors.initials: MX
authors.lastname: Li
authors.forename: J T
authors.initials: JT
authors.lastname: Wu
authors.forename: L
authors.initials: L
authors.lastname: Liu
authors.forename: D M
authors.initials: DM
authors.lastname: Lubman
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary; D004912:Erythrocytes / Q000737:chemistry; D005914:Globins / Q000032:analysis; D006454:Hemoglobins / Q000032:analysis*; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D013056:Spectrophotometry, Ultraviolet
chemical_list: D006454:Hemoglobins; D005914:Globins