Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
38, Number 6, June 2000, pp. 255-258
High-Throughput
Method Development Approaches for Bioanalytical Mass SpectrometryJohn
S. Janiszewski, Monica C. Swyden, and Hassan G. Fouda
Central Research Division, Pfizer Inc., Eastern Point Rd., Groton, CT 06340
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-well 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 ³ 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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