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

Rapid Commun Mass Spectrom. 1997;11(1):99-108. doi: 10.1002/(SICI)1097-0231(19970115)11:1<99::AID-RCM813>3.0.CO;2-6.

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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Chromatography, High Pressure Liquid
  • Electrophoresis, Capillary
  • Erythrocytes / chemistry
  • Globins / analysis
  • Hemoglobins / analysis*
  • Humans
  • Mass Spectrometry
  • Molecular Sequence Data
  • Spectrophotometry, Ultraviolet

Substances

  • Hemoglobins
  • Globins