High-throughput biological sample analysis using on-line turbulent flow extraction combined with monolithic column liquid chromatography/tandem mass spectrometry.
A high-throughput liquid chromatography/tandem mass spectrometry (LC/MS/MS) method, which combines on-line sample extraction through turbulent flow chromatography with a monolithic column separation, has been developed for direct injection analysis of drugs and metabolites in human plasma samples. By coupling a monolithic column into the system as the analytical column, the method enables running 'dual-column' extraction and chromatography at higher flow rates, thus significantly reducing the time required for the transfer and mixing of extracted fraction onto the separation column as well as the time for gradient separation. A strategy of assessing and reducing the matrix suppression effect on the on-line extraction LC/MS/MS has also been discussed. Experiments for evaluating the resolution, peak shape, sensitivity, speed, and matrix effect were conducted with dextromethorphan and its metabolite dextrorphan as model compounds in human plasma matrix. It was demonstrated that the total run time for this assay with a baseline separation of two analytes is less than 1.5 min.
Zhou
Shaolian
S
DMPK-Bioanalytical Chemistry Department, Covance Laboratories Inc., Madison, WI 53704, USA. shaolian.zhou@novartis.com
Zhou
Huiyu
H
Larson
Mike
M
Miller
Dennis L
DL
Mao
Dunmin
D
Jiang
Xiangyu
X
Naidong
Weng
W
D000701:Analgesics, Opioid / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D003915:Dextromethorphan / Q000097:blood*; D003917:Dextrorphan / Q000097:blood*; D018691:Excitatory Amino Acid Antagonists / Q000097:blood*; D006801:Humans; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*
D000701:Analgesics, Opioid; D018691:Excitatory Amino Acid Antagonists; D003917:Dextrorphan; D003915:Dextromethorphan