Microchip electrospray: improvements in spray and signal stability during gradient elution by an inverted postcolumn makeup flow.

Dynamic changes in mobile phase composition during high-performance liquid chromatography (HPLC) gradient elution coupled to mass spectrometry (MS) sensitively affect electrospray modes. We investigate the impact of the eluent composition on spray stability and MS response by infusion and injection experiments with a small tetrapeptide in water-acetonitrile mixtures. The employed HPLC/electrospray (ESI)-MS configuration uses a microchip equipped with an enrichment column, a separation column, and a makeup flow (MUF) channel.

Microchip electrospray: cone-jet stability analysis for water-acetonitrile and water-methanol mobile phases.

Changes in mobile phase composition during high performance liquid chromatography (HPLC) gradient elution coupled to mass spectrometry (MS) sensitively affect electrospray operation modes. In this work, we identify the influences of dynamic changes in bulk conductivity on the cone-jet stability island for aqueous acetonitrile and aqueous methanol mobile phases commonly used in reversed-phase HPLC. Bulk conductivities of the mobile phases were varied by adding different amounts of formic acid.

Microchip electrospray performance during gradient elution with bulk conductivity changes.

This work identifies dynamic changes in bulk conductivity during reversed-phase HPLC gradient elution as a major source for spray mode changes and instabilities observed in ESI-MS. A commercial microchip-HPLC/ESI-MS configuration was modified to enable electrospray diagnostics based on frequency analysis of the microchip emitter current combined with spray imaging. This approach facilitated detection of different spray modes together with their onset potentials.

Chip-based nano-LC-MS/MS identification of proteins in complex biological samples using a novel polymer microfluidic device.

Although the proteome of each organism is unambiguously coded in its genome, the proteome shows the real biology in action in each particular organism. New powerful tools are being developed for biochemists and biologists to analyze complex biological samples for studying the complete protein supplement of the genome, i. e.