Heart-cutting two-dimensional capillary electrophoresis for the on-line purification and separation of derivatized amino acids.
Heart-cutting two-dimensional (2D) capillary electrophoresis (CE) in a single capillary was used for analysis of derivatized amino acids. A mixture of 12 amino acids derivatized with UV-active benzyl 4-(3-(2-chloroethyl)-3-nitrosoureido)butylcarbamate label served as a model of a moderately complex sample due to the presence of numerous derivatization byproducts. The first step of the heart-cutting 2D approach was sample cleanup by capillary zone electrophoresis (CZE) in borate electrolyte. Then, only a selected portion of the first-dimension separation was transferred into the second dimension of the separation by a specific voltage and pressure program. Finally, the zone of derivatized amino acids was separated by micellar electrokinetic chromatography in a borate-sodium dodecyl sulfate system. The whole 2D process can be performed in a conventional CE analyzer without any interface for connection of the two separation modes. Intraday repeatability of the total migration time was 2%. In general, the heart-cutting 2D-CE methodology in a single capillary can be adapted for any CE mode regardless of the direction and velocity of electroosmotic flow and position of the fraction of interest in the first dimension (i.e., first, last, or intermediate fraction).
Anouti
Suzanne
S
Institut des Biomolécules Max Mousseron (UMR 5247 CNRS-Université de Montpellier 1-Université de Montpellier 2), place Eugène Bataillon CC 017, 34095 Montpellier Cedex 5, France.
Vandenabeele-Trambouze
Odile
O
Koval
Dusan
D
Cottet
Hervé
H
D000596:Amino Acids / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D011312:Pressure
D000596:Amino Acids