Detailed study of precipitation of a poorly water soluble test compound using methodologies as in activity and solubility screening - mixing and automation effects

Cheska Gillespie, Alan R. Kennedy, Darren Edwards, Lee Dowden, Pierre Daublain, Peter Halling

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1 Citation (Scopus)

Abstract

Storage of pharmaceutical discovery compounds dissolved in dimethylsulfoxide (DMSO) is commonplace within industry. Often, the DMSO stock solution is added to an aqueous system (e. g. in bioassay or kinetic solubility testing)-since most test compounds are hydrophobic, precipitation could occur. Little is known about the factors affecting this precipitation process at the low (mu M) concentrations used in screening analyses. Here, a poorly water soluble test compound (tolnaftate) was used to compare manual and automated pipetting, and explore the effect of mixing variables on precipitation. The amount of drug present in the supernatant after precipitation and centrifugation of the samples was quantified. An unusual result was obtained in three different laboratories: results of experiments performed initially were statistically significantly higher than those performed after a few days in the same lab. No significant differences were found between automated and manual pipetting, including in variability. Vortex mixing was found to give significantly lower supernatant amounts compared to milder mixing types. The mixing employed affects the particle growth of the precipitate. These findings are of relevance to discovery stage bioassay and kinetic solubility analyses.

Original languageEnglish
Pages (from-to)636-643
Number of pages8
JournalCombinatorial Chemistry and High Throughput Screening
Volume16
Issue number8
Publication statusPublished - Sep 2013

Fingerprint

Automation
Dimethyl Sulfoxide
Biological Assay
Solubility
Screening
Tolnaftate
Water
Bioassay
Centrifugation
Pharmaceutical Preparations
Industry
Kinetics
Growth
Drug products
Precipitates
Vortex flow
Testing
Experiments

Keywords

  • drug screening
  • particle
  • throughput
  • drug precipitation
  • NTA
  • mixing
  • growth
  • discovery
  • promiscuous inhibitors

Cite this

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AU - Gillespie, Cheska

AU - Kennedy, Alan R.

AU - Edwards, Darren

AU - Dowden, Lee

AU - Daublain, Pierre

AU - Halling, Peter

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AB - Storage of pharmaceutical discovery compounds dissolved in dimethylsulfoxide (DMSO) is commonplace within industry. Often, the DMSO stock solution is added to an aqueous system (e. g. in bioassay or kinetic solubility testing)-since most test compounds are hydrophobic, precipitation could occur. Little is known about the factors affecting this precipitation process at the low (mu M) concentrations used in screening analyses. Here, a poorly water soluble test compound (tolnaftate) was used to compare manual and automated pipetting, and explore the effect of mixing variables on precipitation. The amount of drug present in the supernatant after precipitation and centrifugation of the samples was quantified. An unusual result was obtained in three different laboratories: results of experiments performed initially were statistically significantly higher than those performed after a few days in the same lab. No significant differences were found between automated and manual pipetting, including in variability. Vortex mixing was found to give significantly lower supernatant amounts compared to milder mixing types. The mixing employed affects the particle growth of the precipitate. These findings are of relevance to discovery stage bioassay and kinetic solubility analyses.

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KW - discovery

KW - promiscuous inhibitors

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