title: Supercritical fluid chromatography-tandem mass spectrometry for fast bioanalysis of R/S-warfarin in human plasma.
abstract: Chiral separation for the analysis of enantiomers in biological fluids by HPLC often takes relatively long chromatography time compared to achiral analysis. The advantage of fast mass transfer in packed-column supercritical fluid chromatography (pSFC) and the high-flow compatibility of APCI-MS/MS were applied to develop a fast bioanalytical method for R/S-warfarin in human plasma. Presented here are the main challenges encountered during method development of a semi-automated liquid extraction SFC-MS/MS method. The selection of internal standard, robustness of the SFC equipment, and carryover issues are discussed. The method has high-throughput: the chromatography time is at least two-fold faster than the our fastest previous method; and the liquid/liquid extraction time of 96 samples is less than 20 min using a Tecan Genesis RSP 100 pipetting station and a Tomtec Quadra-96 workstation. The standard curve range was 13.6-2500 ng/ml. Precision of QC concentrations from four validation runs was 7.0% for R-warfarin and 6.0% C.V. for S-warfarin; and the bias was 3.7 and 3.2% R.E., respectively. The method is sensitive, accurate, selective and robust, and was applied to a drug-interaction clinical study with rapid turnaround of sample analysis.
authors.affiliation: MDS Pharma Services, 621 Rose Street, Lincoln, NE 68502, USA. roger.coe@mdsinc.com
authors.forename: Roger A
authors.initials: RA
authors.lastname: Coe
authors.forename: Jonathan O
authors.initials: JO
authors.lastname: Rathe
authors.forename: Jean W
authors.initials: JW
authors.lastname: Lee
mesh_terms: D002138:Calibration; D025924:Chromatography, Supercritical Fluid / Q000295:instrumentation / Q000379:methods; D006801:Humans; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D014859:Warfarin / Q000097:blood* / Q000737:chemistry
chemical_list: D014859:Warfarin