Enantiomer separation of amino acids in immunoaffinity micro LC-MS.
Chiral immunoaffinity microbore columns were directly interfaced with MS detection, and the effect of column length and temperature on the enantiomer separation of a number of underivatized aromatic and aliphatic amino acids was investigated utilizing an antibody chiral stationary phase that had been prepared by immobilizing a monoclonal anti-D-amino acid antibody onto silica. The stronger affinity of the antibody towards aromatic and bulky amino acids allowed separation of such analytes in a 0.75 x 150 mm column, while an increase in column length enabled separation of more weakly bound compounds. The strength of interaction between chiral selector and analytes could be modulated conveniently by lowering the temperature. For the first time, simultaneous enantiomer separation of mixtures of amino acids was achieved on antibody-based chiral stationary phases using extracted ion chromatograms.
Zeleke
Jessica M
JM
Department of Chemistry and Biochemistry, Northern Illinois University, DeKalb, Illinois 60115-2862, USA.
Smith
Gregory B
GB
Hofstetter
Heike
H
Hofstetter
Oliver
O
D000596:Amino Acids / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D001639:Bicarbonates / Q000737:chemistry; D002021:Buffers; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry / Q000379:methods*; D008904:Miniaturization; D015394:Molecular Structure; D009994:Osmolar Concentration; D011506:Proteins / Q000737:chemistry / Q000276:immunology; D013237:Stereoisomerism
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D001639:Bicarbonates; D002021:Buffers; D011506:Proteins; C027043:ammonium bicarbonate