Development of an automated mass spectrometry system for the quantitative analysis of liver microsomal incubation samples: a tool for rapid screening of new compounds for metabolic stability

Abstract

There is a continuing need for increased throughput in the evaluation of new drug entities in terms of their pharmacokinetic parameters. One useful parameter that can be measured in vitro using liver microsomal preparations is metabolic stability. In this report, we describe an automated system that can be used for unattended quantitative analysis of liver microsomal samples for a series of compounds. This system is based on the Sciex API 150 (single quadrupole) liquid chromatography/mass spectrometry system and utilizes 96-well plate autosampler technology as well as a custom-designed AppleScript which executes the on-line data processing and report generation. It has the capability of analyzing at least 75 compounds per week or 300 compounds per month in an automated fashion.

The Expanding Role of HPLC in Drug Discovery

“…Manual processing of so many samples would clearly render the data processing and data reporting rate-limiting. To address this, numerous groups have combined the power of vendor software programs that automate peak area determinations with visual basic programming to provide methods for data processing and data reporting [86][87][88]. Shown in Figure 11-13 is a partial summary report of a plate of eight reference compounds and 88 test compounds received from a drug discovery project.…”

Quantitative bioanalysis: an integrated approach for drug discovery and development

“…The correlation between in vivo hepatic clearance values and the intrinsic clearance values determined from liver microsomal incubation experiments is also well documented [3][4][5][6]. The application of this in vitro screening approach consistently generates hundreds of samples for analysis in a single batch, prompting researchers to develop clever analytical techniques involving LC-MS [7][8][9]. Korfmacher et al [7] introduced an automated system for quantitative analysis of metabolic stability samples incorporating LC-MS and automated data processing strategies.…”

“…The application of this in vitro screening approach consistently generates hundreds of samples for analysis in a single batch, prompting researchers to develop clever analytical techniques involving LC-MS [7][8][9]. Korfmacher et al [7] introduced an automated system for quantitative analysis of metabolic stability samples incorporating LC-MS and automated data processing strategies. Samples were analyzed serially using a relatively lengthy HPLC gradient analysis time (10 min).…”

Fast LC/MS in the analysis of small molecules

“…Korfmacher et al [41] created a fast generic gradient method for the quantitative analysis of liver microsomal incubation samples. The method uses a 50 mm×3.9 mm C 18 column at a flow rate of 0.8 mL min -1 over a 7-min gradient run time.…”