High-throughput method development approaches for bioanalytical mass spectrometry.
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 > or = 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.
Janiszewski
J S
JS
Central Research Division, Pfizer Inc., Groton, CT 06340, USA.
Swyden
M C
MC
Fouda
H G
HG
D000097:Acetonitriles; D001826:Body Fluids / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016903:Drug Monitoring; D006801:Humans; D007202:Indicators and Reagents; D013058:Mass Spectrometry / Q000379:methods*; D017942:Nitric Acid; D011786:Quality Control; D012680:Sensitivity and Specificity; D012997:Solvents
D000097:Acetonitriles; D007202:Indicators and Reagents; D012997:Solvents; D017942:Nitric Acid; C032159:acetonitrile