Tandem mass spectrometry with online high-flow reversed-phase extraction and normal-phase chromatography on silica columns with aqueous-organic mobile phase for quantitation of polar compounds in biological fluids.
In this work, high-flow online reversed-phase extraction was coupled with normal phase on silica columns with aqueous-organic mobile phase liquid chromatography/tandem mass spectrometry (LC/MS/MS) to quantify drug candidates in biological fluids. The orthogonal separation effect obtained from this configuration considerably reduced matrix effects and increased sensitivity for highly polar compounds as detected by selected reaction monitoring. This approach also significantly improved the robustness and limit of detection of the assays. An evaluation of this system was performed using a mixture of albuterol and bamethan in rat plasma. Assay validation demonstrated acceptable accuracy (< 8% difference) and precision (< 6% CV) for these model compounds. The system has been used for the quantitation of polar ionic compounds in biological fluids in support of drug discovery programs. This assay was used to analyze samples for a BMS proprietary compound (A) in a rat pharmacokinetic study and is shown as an example to demonstrate the precision, accuracy, and sufficient sensitivity of this system.
Deng
Yuzhong
Y
PCO, Bristol-Myers Squibb Company, P.O. Box 4000, Princeton, NJ 85430, USA. alan.deng@bms.com
Zhang
Hongwei
H
Wu
Jing-Tao
JT
Olah
Timothy V
TV
D000420:Albuterol / Q000097:blood*; D001774:Blood Chemical Analysis / Q000379:methods*; D001826:Body Fluids / Q000737:chemistry*; D005591:Chemical Fractionation / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D004983:Ethanolamines / Q000097:blood*; D044085:Microfluidics / Q000379:methods*; D009862:Online Systems; D009930:Organic Chemicals / Q000737:chemistry; D044367:Phase Transition; D010627:Phenethylamines / Q000097:blood*; D012822:Silicon Dioxide / Q000737:chemistry; D055672:Static Electricity; D014867:Water / Q000737:chemistry
D004983:Ethanolamines; D009930:Organic Chemicals; D010627:Phenethylamines; D014867:Water; D012822:Silicon Dioxide; D000420:Albuterol; C084810:bamethan