Development of field modulation in a split-field drift tube for high-throughput multidimensional separations.
A field modulation approach for high-throughput ion mobility/time-of-flight analyses of complex mixtures has been developed using a split-field drift tube. In this approach, complex mixtures of peptides, such as those that arise from tryptic digestion of protein mixtures, are separated by nanocolumn liquid chromatography, ionized by electrospray ionization, and analyzed by ion mobility/time-of-flight techniques. The split-field drift tube allows parent ions to be separated based on differences in their low-field mobilities through the first-field region before entering the second region. For increased throughput, the magnitude of the field in the second region can be modulated throughout an LC separation in order to favor transmission of different types of ions: parent ions at low fields; fragments from primarily [M+3H]3+ peptides at moderate fields; or, fragmentation of [M+3H]3+ and [M+2H]2+ species at higher fields. We demonstrate the approach with two examples: a mixture of tryptic peptides from digestion of hemoglobin; and a complex mixture of tryptic peptides from digestion of human plasma.
Koeniger
Stormy L
SL
Department of Chemistry, Indiana University, Bloomington, Indiana 47405, USA.
Valentine
Stephen J
SJ
Myung
Sunnie
S
Plasencia
Manolo
M
Lee
Young Jin
YJ
Clemmer
David E
DE
D001798:Blood Proteins / Q000032:analysis / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D006454:Hemoglobins / Q000032:analysis / Q000378:metabolism; D006801:Humans; D008722:Methods; D010446:Peptide Fragments / Q000032:analysis; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation*; D014357:Trypsin / Q000378:metabolism
D001798:Blood Proteins; D006454:Hemoglobins; D010446:Peptide Fragments; D014357:Trypsin