Automated solid-phase microextraction and thin-film microextraction for high-throughput analysis of biological fluids and ligand-receptor binding studies.
This protocol describes how to perform automated solid-phase microextraction (SPME) and thin-film microextraction (TFME) in a 96-well plate format for high-throughput analysis of drugs, metabolites and any other analytes of interest in biological fluids using liquid chromatography-electrospray tandem mass spectrometry. Sample preparation time required is typically 1 min per sample; hence, the throughput achievable with automated SPME/TFME is comparable with automated 96-well liquid-liquid extraction and solid-phase extraction methods, but greater than most online solid-phase extraction methods. The technique is applicable to complex samples such as whole blood without additional pretreatment. The amount of analyte extracted by SPME/TFME is proportional to the free (unbound) concentration of the analyte; hence, SPME/TFME can be used to determine both total and free concentrations of analytes from a single biofluid sample and to perform automated ligand-receptor binding studies in order to determine binding affinity and/or overall extent of ligand binding to a complex biofluid.
Vuckovic
Dajana
D
Department of Chemistry, University of Waterloo, Waterloo, Ontario, Canada.
Cudjoe
Erasmus
E
Musteata
Florin Marcel
FM
Pawliszyn
Janusz
J
D001331:Automation; D001569:Benzodiazepines / Q000737:chemistry; D001774:Blood Chemical Analysis / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D011485:Protein Binding; D052617:Solid Phase Microextraction / Q000295:instrumentation / Q000379:methods*; D014556:Urine / Q000737:chemistry*
D008024:Ligands; D001569:Benzodiazepines