Supercritical fluid chromatography-tandem mass spectrometry for fast bioanalysis of R/S-warfarin in human plasma.
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.
Coe
Roger A
RA
MDS Pharma Services, 621 Rose Street, Lincoln, NE 68502, USA. roger.coe@mdsinc.com
Rathe
Jonathan O
JO
Lee
Jean W
JW
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
D014859:Warfarin