Development and Testing of a Colorimetric 96 Well Plate Assay for the Determination of HD Hydrolysis Rate in Various Formulations
Abstract
Previous aqueous studies using HD pre-dissolved in isopropanol have shown that the effectiveness of HD enzymatic degradation is a function of the homogeneity of the HD-water system. In this study, a microtiter plate assay was developed for the purpose of screening a series of compounds with the potential to increase the solubility or dispersion of HD in an aqueous matrix. The assay used meta-cresol purple dye as a pH indicator in a series of buffer concentrations to monitor the acid produced from HD hydrolysis. The extent of hydrolysis could be observed colorimetrically in a time-controlled series of reactions that allowed simultaneous comparison of numerous compounds or conditions on a single microtiter plate. Sixty-seven different detergents, surfactants, or different concentrations thereof were screened to determine their effect on HD hydrolysis rate. All were observed to inhibit HD hydrolysis. The simplest explanation of the results observed was probably that the micelles effectively sequestered the HD molecules from water in a hydrophobic environment.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2005
- Accession Number
- ADA431980
Entities
People
- Joseph J. Defrank
- Steven P. Harvey
Organizations
- Edgewood Chemical Biological Center