title: Detailed study of precipitation of a poorly water soluble test compound using methodologies as in activity and solubility screening - mixing and automation effects.
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 (µM) 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.
authors.affiliation: WestCHEM, The Department of Pure and Applied Chemistry, University of Strathclyde, Thomas Graham Building, 295 Cathedral Street, Glasgow, G1 1XL, UK. cheska.gillespie@strath.ac.uk
authors.forename: Cheska
authors.initials: C
authors.lastname: Gillespie
authors.forename: Alan R
authors.initials: AR
authors.lastname: Kennedy
authors.forename: Darren
authors.initials: D
authors.lastname: Edwards
authors.forename: Lee
authors.initials: L
authors.lastname: Dowden
authors.forename: Pierre
authors.initials: P
authors.lastname: Daublain
authors.forename: Peter
authors.initials: P
authors.lastname: Halling
mesh_terms: D000935:Antifungal Agents / Q000737:chemistry*; D011232:Chemical Precipitation; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D012995:Solubility; D014047:Tolnaftate / Q000737:chemistry*; D014867:Water / Q000737:chemistry*
chemical_list: D000935:Antifungal Agents; D014867:Water; D014047:Tolnaftate; D004121:Dimethyl Sulfoxide