High-Throughput Method Development Approaches for Bioanalytical Mass Spectrometry

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Abstract
A rational approach to the development and optimization of solid-phase extraction (SPE) methods is described. The semiautomated scheme allows for the simultaneous testing of multiple chemistries using a custom multiple-sorbent 96-we 11 method development plate. Optimized extraction conditions for up to five analytes are determined in a single 2.5-h experiment. The experiment can be tailored to determine SPE conditions (including wash protocols) for related analytes. Data obtained by liquid chromatography-atmospheric pressure ionization-mass spectrometry allows the quantitation of absolute recovery and selection of the best extraction conditions for approximately 100 analytes of diverse structure. Optimized extraction protocols yielding at least 80% recovery are determined for 81 % of the analytes. For 96% of the analytes screened, extraction conditions resulting in recoveries of ≥ 60% are determined. The most generic set of SPE conditions consist of either C8 or C18 sorbent with an eluent composition of acetonitrile with 5mM nitric acid added.

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