title: Simultaneous LC-MS/MS quantitation of a highly hydrophobic pharmaceutical compound and its metabolite in urine using online monolithic phase-based extraction.
abstract: This article describes the use of monolithic material as extraction support in simultaneous LC-MS/MS quantitation of a highly hydrophobic pharmaceutical compound and its hydroxylated metabolite in urinary matrix. DMSO was added to urine samples to aid the solubilization of analytes during storage and transfer. Samples were then diluted and injected onto an online extraction system for high-throughput analysis in an automated fashion. A linear range of 1.03-103 ng/mL was validated for the parent compound and 7.02-1400 ng/mL for the metabolite. The accuracy (%bias) at the lower LOQ (LLOQ) for the parent compound was -2.9% and the precision (%CV) at the LLOQ was 5.4%, while the accuracy at LLOQ for the metabolite was 6.3% and the precision at LLOQ was 5.5%. The results show that quantitative LC-MS/MS analysis by online extraction using the monolithic support is reproducible, sensitive, and accurate enough to satisfy bioanalytical testing requirements in a good laboratory practice (GLP) regulated environment. Monolithic phase based online extraction provided efficient sample cleanup, which was evidenced in matrix effect testing against multiple lots of urinary matrix. The method described within can be a generic approach for quantitative analysis of analytes with poor solubility in aqueous matrix.
authors.affiliation: Department of Drug Analysis, Abbott Laboratories, Abbott Park, IL 60064-6126, USA. Raymond.Xu@abbott.com
authors.forename: Raymond Naxing
authors.initials: RN
authors.lastname: Xu
authors.forename: Brian
authors.initials: B
authors.lastname: Boyd
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Rieser
authors.forename: Tawakol A
authors.initials: TA
authors.lastname: El-Shourbagy
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000378:metabolism / Q000652:urine; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000295:instrumentation* / Q000379:methods*; D013499:Surface Properties; D053719:Tandem Mass Spectrometry / Q000295:instrumentation* / Q000379:methods*; D013997:Time Factors
chemical_list: D004364:Pharmaceutical Preparations