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. One nano pump is connected to the separation column, while a second one delivers solvent of exactly inverted composition to the MUF channel. Both solvent streams are united behind the separation column, before the ESI tip, such that the resulting electrosprayed solution always has identical composition during a gradient elution. Analyte peak parameters without and with MUF compensation are determined and discussed with respect to the electrospray mode and eluent composition. The postcolumn MUF significantly improves spray and signal stability over the entire solvent gradient, without compromising the performance of the HPLC separation column. It can also be conveniently implemented on microchip platforms.
Department of Chemistry, Philipps-Universität Marburg, Hans-Meerwein-Strasse, 35032 Marburg, Germany.
Stephanie
S
Jung
Uwe
U
Effelsberg
Ulrich
U
Tallarek
D000097:Acetonitriles / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D046208:Microchip Analytical Procedures; D009842:Oligopeptides / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*; D014867:Water / Q000737:chemistry
D000097:Acetonitriles; D009842:Oligopeptides; D014867:Water; C032159:acetonitrile