Coupling of hydrodynamically closed large bore capillary isotachophoresis with electrospray mass spectrometry.

Capillary isotachophoresis with coupled columns provides efficient means for rapid electrophoretic analysis of sample volumes of up to 10microl or more. Commercially available instruments are commonly equipped with conductivity and UV absorbance detectors; however, their on-line coupling with electrospray mass spectrometry is highly desirable. In this work we have modified the commercial coupled column isotachophoresis system for direct connection to an ion trap mass spectrometer. The design included attachment of an elution block with a short capillary transfer line directing the separated zones towards the mass spectrometer. The modification further included separation of the injection and electrode blocks from the separation columns by semipermeable membranes eliminating unwanted fluid movements in the wide bore fluoropolymer separation capillaries. Fused silica capillaries with varying internal diameter were connected as a transfer line between the elution block and mass spectrometer. The transfer line served also as the ESI tip of the sheathless electrospray interface. During the analysis the first, wide bore preseparation capillary with 0.8mm internal diameter served for removal of the bulk sample components and preseparation of the potentially interfering analytes. After the electronic column switching the separation was finished in a 0.3mm internal diameter capillary and the separated ITP zones were transferred in-line by a spray liquid towards the mass spectrometer. The instrumentation was tested for determination of vitamins in whole blood analysis and separation of tryptic peptides.