title: Performance of an ultra-low elution-volume 96-well plate: drug discovery and development applications.
abstract: Recently, sample preparation has been considered to be the major cause of bottlenecks during high-throughput analysis. With the assistance of robotic liquid handlers and the 96-well plate format, more samples can be prepared for subsequent liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis. Protein precipitation is still widely used despite potential loss of sensitivity or variable results due to ion suppression. The use of solid-phase extraction (SPE) clearly gives superior results but may not be as cost effective as protein precipitation due to the labor and material costs associated with the process. Here, a novel 96-well SPE plate is described that was designed to minimize the elution volume required for quantitative elution of analytes. The plate is packed with 2 mg of a high-capacity SPE sorbent that allows loading of up to 750 microL of plasma, while the novel design permits elution with as little as 25 microL. Therefore, the plate offers up to a 15-fold increase in sample concentration. The evaporation and reconstitution step that is typically required in SPE is avoided due to the concentrating ability of the plate. Examples of applications in drug discovery/development are shown and results are compared to protein precipitation. Excellent sensitivity and linearity are demonstrated.
authors.affiliation: Waters Corporation, 34 Maple St., Milford, MA 01757-3696, USA. claude_mallet@waters.com
authors.forename: Claude R
authors.initials: CR
authors.lastname: Mallet
authors.forename: Ziling
authors.initials: Z
authors.lastname: Lu
authors.forename: Ray
authors.initials: R
authors.lastname: Fisk
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Mazzeo
authors.forename: Uwe D
authors.initials: UD
authors.lastname: Neue
mesh_terms: D000639:Amitriptyline / Q000097:blood; D000818:Animals; D002138:Calibration; D011232:Chemical Precipitation; D002853:Chromatography, Liquid / Q000295:instrumentation*; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods; D051379:Mice; D008832:Microchemistry / Q000295:instrumentation; D004364:Pharmaceutical Preparations / Q000097:blood*; D011433:Propranolol / Q000097:blood; D011506:Proteins / Q000737:chemistry; D051381:Rats; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation*
chemical_list: D004364:Pharmaceutical Preparations; D011506:Proteins; D000639:Amitriptyline; D011433:Propranolol