The Determination of Drug Protein Binding by HPLC Using a Chemically Bonded Bovine-Albumin Stationary Phase
The binding of drugs to proteins may play an important role in drug bioavailability and in interactions with other drugs bound to proteins. Determination of drug protein binding by equilibrium dialysis  is rather time-consuming; size-exclusion chromatography has been described as an alternative , but special precautions are required to prevent dissociation of the drug-protein comples . Yoshida et al.  used proteins physically bonded to ODS (C-18) columns to separate drugs. Here we describe the use of a commercially available chemically bonded bovine-albumin stationary phase to determine the degree of protein binding of potential drugs. To obtain the required hydrophobic interaction a buffer concentration of ~0.1 M is necessary, and the addition of 1-propanol lowers retention for all solutes . The retention characteristics of a heterogeneous group of 23 compounds and a group of 11 analogous potential drugs has now been studied. We compared the resulting data with equilibrium dialysis results. The amount injected for chromatography had to be small (~0.5 pmol) to prevent overloading of the Chromatographic system.
KeywordsSulfa Compound Potential Drug Buffer Concentration retentIon Volume Equilibrium Dialysis
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