title: Increasing bioanalytical throughput using pcSFC-MS/MS: 10 minutes per 96-well plate.
abstract: The utility of packed-column supercritical, subcritical, and enhanced fluidity liquid chromatographies (pcSFC) for high-throughput applications has increased during the past few years. In contrast to traditional reversed-phase liquid chromatography, the addition of a volatile component to the mobile phase, such as CO2, produces a lower mobile-phase viscosity. This allows the use of higher flow rates which can translate into faster analysis times. In addition, the resulting mobile phase is considerably more volatile than the aqueous-based mobile phases that are typically used with LC-MS, allowing the entire effluent to be directed into the MS interface. High-throughput bioanalytical quantitation using pcSFC-MS/MS for pharmacokinetics applications is demonstrated in this report using dextromethorphan as a model compound. Plasma samples were prepared by automated liquid/liquid extraction in the 96-well format prior to pcSFC-MS/MS analysis. Three days of validation data are provided along with study sample data from a patient dosed with commercially available Vicks 44. Using pcSFC and MS/MS, dextromethorphan was quantified in 96-well plates at a rate of approximately 10 min/plate with average intraday accuracy of 9% or better. Daily relative standard deviations (RSDs) were less than 10% for the 2.21 and 14.8 ng/mL quality control (QC) samples, while the RSDs were less than 15% at the 0.554 ng/mL QC level.
authors.affiliation: Health Care Research Center, The Procter & Gamble Company, Mason, Ohio 45040, USA. hoke.sh@pg.com
authors.forename: S H
authors.initials: SH
authors.lastname: Hoke
authors.forename: J A
authors.initials: JA
authors.lastname: Tomlinson
authors.forename: R D
authors.initials: RD
authors.lastname: Bolden
authors.forename: K L
authors.initials: KL
authors.lastname: Morand
authors.forename: J D
authors.initials: JD
authors.lastname: Pinkston
authors.forename: K R
authors.initials: KR
authors.lastname: Wehmeyer
mesh_terms: D000996:Antitussive Agents / Q000097:blood* / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D003915:Dextromethorphan / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000996:Antitussive Agents; D003915:Dextromethorphan