Automated solid-phase extraction workstations combined with quantitative bioanalytical LC/MS.
An automated solid-phase extraction workstation was used to develop, characterize and validate an LC/MS/MS method for quantifying a novel lipid-regulating drug in dog plasma. Method development was facilitated by workstation functions that allowed wash solvents of varying organic composition to be mixed and tested automatically. Precision estimates for this approach were within 9.8% relative standard deviation (RSD) across the calibration range. Accuracy for replicate determinations of quality controls was between -7.2 and +6.2% relative error (RE) over 5-1,000 ng/ml(-1). Recoveries were evaluated for a wide variety of wash solvents, elution solvents and sorbents. Optimized recoveries were generally > 95%. A sample throughput benchmark for the method was approximately equal 8 min per sample. Because of parallel sample processing, 100 samples were extracted in less than 120 min. The approach has proven useful for use with LC/MS/MS, using a multiple reaction monitoring (MRM) approach.
Huang
N H
NH
Department of Pharmacokinetics and Dynamics, Metabolism, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert, Ann Arbor, MI 48105, USA.
Kagel
J R
JR
Rossi
D T
DT
D000818:Animals; D002138:Calibration; D002208:Caproates / Q000097:blood; D002853:Chromatography, Liquid / Q000379:methods*; D004285:Dogs; D005069:Evaluation Studies as Topic; D000960:Hypolipidemic Agents / Q000097:blood; D013058:Mass Spectrometry / Q000379:methods*; D015203:Reproducibility of Results; D012371:Robotics / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D012997:Solvents; D013997:Time Factors
D002208:Caproates; D000960:Hypolipidemic Agents; D012997:Solvents; C111024:gemcabene