An integrated platform for fully automated high-throughput LC–MS/MS analysis of in vitro metabolic stability assay samples

Luippold, Andreas H.; Arnhold, Thomas; Jörg, Wolfgang; Süssmuth, Roderich D.

doi:10.1016/j.ijms.2010.07.004

Cited by 45 publications

(45 citation statements)

References 38 publications

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“…An additional advantage that RF-MS/MS enjoys is that it requires either no or very minimal changes in sample preparation as compared to LC-MS/MS analysis, making it easy to be incorporated into existing automated in vitro assays. [19] To date this technology has been successfully used to support several high-throughput screening assays, [20,21] cytochrome P450 (CYP) inhibition assays, [22,23] as well as a metabolic stability assay, [24] and in this manuscript we describe the development of a sensitive, selective, reproducible and robust RF-MS/MS method for digoxin bioanalysis to support an in vitro P-gp inhibition screen.…”

mentioning

confidence: 99%

Ultrafast mass spectrometry based bioanalytical method for digoxin supporting an in vitro P‐glycoprotein (P‐gp) inhibition screen

Wagner

Kolb

Özbal³

et al. 2011

Rapid Comm Mass Spectrometry

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The evaluation of interactions between drug candidates and transporters such as P-glycoprotein (P-gp) has gained considerable interest in drug discovery and development. Inhibition of P-gp can be assessed by performing bi-directional permeability studies with in vitro P-gp-expressing cellular model systems such as Caco-2 (human colon carcinoma) cells, using digoxin as a substrate probe. Existing methodologies include either assaying (3)H-digoxin with liquid scintillation counting (LSC) detection or assaying non-labeled digoxin with liquid chromatography-tandem mass spectrometric (LC-MS/MS) analysis at a speed of several minutes per sample. However, it is not feasible to achieve a throughput high enough using these approaches to sustain an early liability screen that generates more than a thousand samples on a daily basis. To address this challenge, we developed an ultrafast (9 s per sample) bioanalytical method for digoxin analysis using RapidFire™, an on-line solid-phase extraction (SPE) system, with MS/MS detection. A stable isotope labeled analog, d3-digoxin, was used as internal standard to minimize potential ionization matrix effect during the RF-MS/MS analysis. The RF-MS/MS method was more than 16 times faster than the LC-MS/MS method but demonstrated similar sensitivity, selectivity, reproducibility, linearity and robustness. P-gp inhibition results of multiple validation compounds obtained with this RF-MS/MS method were in agreement with those generated by both the LC-MS/MS method and the (3)H-radiolabel assay. This method has been successfully deployed to assess P-gp inhibition potential as an important early liability screen for drug-transporter interaction.

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mentioning

confidence: 99%

Ultrafast mass spectrometry based bioanalytical method for digoxin supporting an in vitro P‐glycoprotein (P‐gp) inhibition screen

Wagner

Kolb

Özbal³

et al. 2011

Rapid Comm Mass Spectrometry

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“…13,14 in this contribution, we extend and demonstrate the applicability of the automated system to further key aDme assays such as caco-2, pampa, and cYp, including assay type recognition and ms compound optimization.…”

Section: Resultsmentioning

confidence: 88%

“…however, our current efforts are directed to an extension toward the analysis of pharmacokinetic and toxicokinetic in vivo samples. 14 We are aware that the rias represents an in-house customized solution of commercially available components from different vendors. this customization, however, is in accordance with some basic but mandatory rules that can be transferred to any other platform.…”

Section: Pampa-parallel Artificial Membrane Permeation Assay (Pmp)mentioning

confidence: 99%

Application of a Rapid and Integrated Analysis System (RIAS) as a High-Throughput Processing Tool for In Vitro ADME Samples by Liquid Chromatography/Tandem Mass Spectrometry

et al. 2011

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Over the past decade, drug discovery programs have started to address the optimization of key ADME properties already at an early stage of the process. Hence, analytical chemists have been confronted with tremendously rising sample numbers and have had to develop methodologies accelerating quantitative liquid chromatography/tandem mass spectrometry (LC/MS/MS). This article focuses on the application of a generic and fully automated LC/MS/MS, named Rapid and Integrated Analysis System (RIAS), as a high-throughput platform for the rapid quantification of drug-like compounds in various in vitro ADME assays. Previous efforts were dedicated to the setup and feasibility study of a workflow-integrated platform combining a modified high-throughput liquid handling LC/MS/MS system controlled by a customized software interface and a customized data-processing and reporting tool. Herein the authors present an extension of this previously developed basic application to a broad set of ADME screening campaigns, covering CYP inhibition, Caco-2, and PAMPA assays. The platform is capable of switching automatically between various ADME assays, performs MS compound optimization if required, and provides a speed of 8 s from sample to sample, independently of the type of ADME assay. Quantification and peak review are adopted to the high-throughput environment and tested against a standard HPLC-ESI technology.

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“…18 Use of the stepwise synthesis as outlined by Gomaa, and also our one-pot modification (without isolation of the unusually stable quinone-methides), afforded a complex mixture of products (using DMF as solvent) that were each identified either by isolation or by Collision Induced Dissociation (CID) studies of negative ion electrospray mass spectrometry. In the first part of the synthesis, upon addition of one equivalent of triethylamine to the chloranil/malononitrile solution in either methanol or ethanol, it was noted that the pale green color rapidly transformed to an intense blue species, suggesting formation of charge transfer complexes or radical formation.…”

Section: Resultsmentioning

confidence: 99%

“…1 Our strategy has been to identify compounds that initiate antiparasitic drug action from distonic anion radical cascades, 17 using an automated, non-biological screen that could be converted into a high-throughput mode using robotic auto-samplers. 18 We have previously reported that pulsed radiolysis 19 and subsequent negative ion electrospray mass spectrometry 20 were ideally suited to this task and have now identified selected cinnolines as potential antiparasitic agents. 21 In general, densely substituted cinnolines are difficult to construct in a reproducible manner, and to the best of our knowledge, one-pot methods from commercially available raw materials are rare.…”

mentioning

confidence: 99%

One-pot synthesis and negative ion mass spectrometric investigation of a densely functionalized cinnoline

Lambert

Parikh

Messham

et al. 2015

Tetrahedron Letters

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One-pot synthesis and negative ion mass spectrometric investigation of a densely functionalized cinnoline, 3-amino-5,7,8-trichloro-6-hydroxycinnoline-4-carbonitrile http://researchonline.ljmu.ac.uk/2433/ Article LJMU has developed LJMU Research Online for users to access the research output of the University more effectively. Copyright © and Moral Rights for the papers on this site are retained by the individual authors and/or other copyright owners. Users may download and/or print one copy of any article(s) in LJMU Research Online to facilitate their private study or for non-commercial research. You may not engage in further distribution of the material or use it for any profit-making activities or any commercial gain.The version presented here may differ from the published version or from the version of the record. Please see the repository URL above for details on accessing the published version and note that access may require a subscription.

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An integrated platform for fully automated high-throughput LC–MS/MS analysis of in vitro metabolic stability assay samples

Cited by 45 publications

References 38 publications

Ultrafast mass spectrometry based bioanalytical method for digoxin supporting an in vitro P‐glycoprotein (P‐gp) inhibition screen

Ultrafast mass spectrometry based bioanalytical method for digoxin supporting an in vitro P‐glycoprotein (P‐gp) inhibition screen

Application of a Rapid and Integrated Analysis System (RIAS) as a High-Throughput Processing Tool for In Vitro ADME Samples by Liquid Chromatography/Tandem Mass Spectrometry

One-pot synthesis and negative ion mass spectrometric investigation of a densely functionalized cinnoline

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