Ultra-performance liquid chromatography coupled to time-of-flight mass spectrometry for robust, high-throughput quantitative analysis of an automated metabolic stability assay, with simultaneous determination of metabolic data.
The application of ultra-performance liquid chromatography coupled to time-of-flight mass spectrometry (UPLC/TOFMS) for high-throughput analysis of a 96-well plate based metabolic stability assay has been investigated. Full-scan data were acquired, with run times of 2.5-3.5 min, from which narrow window extracted ion chromatograms were generated, producing quantitative data for the test compound equivalent to that obtained by high-performance liquid chromatography with tandem mass spectrometric detection on a triple quadrupole instrument (HPLC/MS/MS). Sensitivity and mass accuracy were maintained over the analysis of >300 samples. Additionally, the UPLC/TOFMS datasets obtained gave access to metabolic route information, at no cost in terms of sensitivity for the test compound.
Merck Sharp & Dohme Research Laboratories, Department of Medicinal Chemistry (Drug Metabolism Section), Neuroscience Research Centre, Terlings Park, Eastwick Road, Harlow, Essex CM20 2QR, UK. desmond_oconnor@merck.com
Desmond
D
O'Connor
Russell
R
Mortishire-Smith
Denise
D
Morrison
Amy
A
Davies
Maria
M
Dominguez
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003975:Diazepam / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004355:Drug Stability; D004743:Enkephalin, Leucine / Q000378:metabolism*; D007099:Imipramine / Q000378:metabolism*; D018377:Neurotransmitter Agents / Q000378:metabolism*; D011619:Psychotropic Drugs / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D018377:Neurotransmitter Agents; D011619:Psychotropic Drugs; D004743:Enkephalin, Leucine; D007099:Imipramine; D003975:Diazepam