Performance of capillary gel electrophoretic analysis of oligonucleotides coupled on-line with electrospray mass spectrometry.
Synthetic oligonucleotides (ODNs) are routinely analyzed using capillary gel electrophoresis (CGE) for size-sieving based separations as well as electrospray mass spectrometry (ESI-MS) for identification. On-line coupling of these methods is therefore desired in order to combine the analytical capabilities provided by both methods. Performance of on-line CGE-ESI-MS systems is influenced by various parameters, and choice of optimal conditions is crucial for successful coupling experiments. In this study, we explore characteristics of the on-line coupled CGE-ESI-MS system for ODN analysis. Effects of CGE buffer concentration, capillary length, separation and orifice voltage on CGE separation and MS detection of a phosphodiester ODN mixture were examined. Attention was paid to the influence of the interface, such as geometry of capillary alignment, sheath liquid flow-rate and sheath liquid composition on performance of the system.
von Brocke
Alexander
A
Research Center for Nucleic Acid and Peptide Chemistry, Institute of Organic Chemistry, University of Tübingen, Auf der Morgenstelle 18, 72076 Tübingen, Germany. alexander.von.brocke@uni-tuebingen.de
Freudemann
Thomas
T
Bayer
Ernst
E
D000097:Acetonitriles / Q000737:chemistry; D000704:Analysis of Variance; D002021:Buffers; D004573:Electrolytes / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D017022:Flow Injection Analysis; D005782:Gels; D009841:Oligonucleotides / Q000032:analysis*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*; D013997:Time Factors
D000097:Acetonitriles; D002021:Buffers; D004573:Electrolytes; D005782:Gels; D009841:Oligonucleotides; C032159:acetonitrile