TAILIEUCHUNG - Make-up Solvent Considerations for Prep and Semi-Prep LC/MS Mass-Based ....

systems utilizing a make-up solvent technique to provide required dilution. . Fraction Collector, 322 H2 Prep Pump, 307 Make-up Pump, 155 UV/VIS Dual . | Make-up Solvent Considerations for Prep and Semi-Prep LC/MS Mass-Based Fraction Collection Application Note 206 Tim Hegeman (Gilson, Inc.) Introduction Solvent choices for electrospray ionization can be separated from those of column separation for systems utilizing a make-up solvent technique to provide required dilution. The following application note takes a look at a few common solvent systems and their effects on a set of pharmaceutical target compounds and related analytes. Materials & Methods What was Investigated Mass-based fraction collection solvent selection for electrospray positive ionization mode was tested for a limited set of solvents against a small set of analytes to show examples of solvent choice on results. Often in prep analysis, unlike analytical work, compromises are made in ionization mode and HPLC conditions to suit a broad number of compounds. In this Application Note, we will look at some of these compromises in terms of ionization. Main Components: Gilson-Thermo Finnigan NEBULA™ Prep System, consisting of a Gilson 215 Liquid Handler/Autosampler/Fraction Collector, 322 H2 Prep Pump, 307 Make-up Pump, 155 UV/VIS Dual- wavelength Detector with flow cell, and Thermo Finnigan MSQ Mass Spectrometer. Experimental—HPLC/LCMS Six solutions of analytes were prepared (10 mg/mL of each compound) in DMSO, which offers at least moderate separation for most analytes. All injection volumes were 500 µL on a 2-mL loop. Column: Metachem 21*50 ODS2. Flow rate: 20 mL/min.; gradient: 5% MEOH/95%Water + HCOOH to 95% MEOH/5% Water + in 10 minutes. Retention time estimates from screening analysis on a ZORBAX *50-mm column at mL/min. using the same gradient. Make-up Flow Rate: mL/min. Splitter: Standard LC-Packings/standard ACM 10–50; UV = 254nm. MS Conditions: Electrospray positive ionization mode. Capillary voltage: 3kV; cone voltage varied for experimental purposes. Capillary temperature (525°C) was determined to give optimal .

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