Driving efficiency in a high-throughput metabolic stability assay through a generic high-resolution accurate mass method and automated data mining

Improving analytical throughput is the focus of many quantitative workflows being developed for early drug discovery. For drug candidate screening, it is common practice to use ultra-high performance liquid chromatography (U-HPLC) coupled with triple quadrupole mass spectrometry. This approach certainly results in short analytical run time; however, in assessing the true throughput, all aspects of the workflow needs to be considered, including instrument optimization and the necessity to re-run samples when information is missed. Here we describe a high-throughput metabolic stability assay with a simplified instrument set-up which significantly improves the overall assay efficiency. In addition, as the data is acquired in a non-biased manner, high information content of both the parent compound and metabolites is gathered at the same time to facilitate the decision of which compounds to proceed through the drug discovery pipeline.

[1]  Wilson Z Shou,et al.  A novel approach to perform metabolite screening during the quantitative LC-MS/MS analyses of in vitro metabolic stability samples using a hybrid triple-quadrupole linear ion trap mass spectrometer. , 2005, Journal of mass spectrometry : JMS.

[2]  Sean Yu,et al.  Quantitation of small molecules using high-resolution accurate mass spectrometers - a different approach for analysis of biological samples. , 2009, Rapid communications in mass spectrometry : RCM.

[3]  A. Makarov,et al.  The Orbitrap: a new mass spectrometer. , 2005, Journal of mass spectrometry : JMS.

[4]  Jennifer H Granger,et al.  Increasing throughput and information content for in vitro drug metabolism experiments using ultra-performance liquid chromatography coupled to a quadrupole time-of-flight mass spectrometer. , 2005, Rapid communications in mass spectrometry : RCM.

[5]  L. Romanyshyn,et al.  Liquid chromatography/tandem mass spectrometric quantification with metabolite screening as a strategy to enhance the early drug discovery process. , 2002, Rapid communications in mass spectrometry : RCM.

[6]  Sean Yu,et al.  High-throughput, accurate mass liquid chromatography/tandem mass spectrometry on a quadrupole time-of-flight system as a 'first-line' approach for metabolite identification studies. , 2008, Rapid communications in mass spectrometry : RCM.

[7]  C. Bigogno,et al.  New perspectives in bio-analytical techniques for preclinical characterization of a drug candidate: UPLC-MS/MS in in vitro metabolism and pharmacokinetic studies. , 2007, Journal of pharmaceutical and biomedical analysis.

[8]  Wilson Z Shou,et al.  Complete profiling and characterization of in vitro nefazodone metabolites using two different tandem mass spectrometric platforms. , 2007, Rapid communications in mass spectrometry : RCM.

[9]  Ari Tolonen,et al.  Comparison of triple quadrupole, hybrid linear ion trap triple quadrupole, time-of-flight and LTQ-Orbitrap mass spectrometers in drug discovery phase metabolite screening and identification in vitro--amitriptyline and verapamil as model compounds. , 2010, Rapid communications in mass spectrometry : RCM.

[10]  Scott Peterman,et al.  An integrated method for metabolite detection and identification using a linear ion trap/Orbitrap mass spectrometer and multiple data processing techniques: application to indinavir metabolite detection. , 2008, Journal of mass spectrometry : JMS.

[11]  Austin C Li,et al.  Two-injection workflow for a liquid chromatography/LTQ-Orbitrap system to complete in vivo biotransformation characterization: demonstration with buspirone metabolite identification. , 2009, Rapid communications in mass spectrometry : RCM.

[12]  A. Nomeir,et al.  Utility of mass spectrometry for in-vitro ADME assays. , 2006, Current drug metabolism.

[13]  E. Nägele,et al.  Simultaneous determination of metabolic stability and identification of buspirone metabolites using multiple column fast liquid chromatography time-of-flight mass spectrometry. , 2007, Journal of chromatography. A.

[14]  Wilson Z Shou,et al.  Simultaneously quantifying parent drugs and screening for metabolites in plasma pharmacokinetic samples using selected reaction monitoring information-dependent acquisition on a QTrap instrument. , 2005, Rapid communications in mass spectrometry : RCM.

[15]  Olavi Pelkonen,et al.  Liquid chromatography-mass spectrometry in in vitro drug metabolite screening. , 2009, Drug discovery today.

[16]  Jie Chen,et al.  Metabolite identification by data-dependent accurate mass spectrometric analysis at resolving power of 60,000 in external calibration mode using an LTQ/Orbitrap. , 2007, Rapid communications in mass spectrometry : RCM.

[17]  Desmond O'Connor,et al.  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. , 2006, Rapid communications in mass spectrometry : RCM.

[18]  Li Di,et al.  Drug-like property concepts in pharmaceutical design. , 2009, Current pharmaceutical design.

[19]  C E Hop,et al.  Integrating qualitative and quantitative liquid chromatography/tandem mass spectrometric analysis to support drug discovery. , 1999, Rapid communications in mass spectrometry : RCM.

[20]  Hongying Gao,et al.  Method for rapid metabolite profiling of drug candidates in fresh hepatocytes using liquid chromatography coupled with a hybrid quadrupole linear ion trap. , 2007, Rapid communications in mass spectrometry : RCM.