The binding of warfarin, a series of non-steroidal anti-inflammatory drugs and a series of benzodiazepines to rat serum albumin (RatSA) and rabbit serum albumin (RabSA) was compared with their binding to human serum albumin (HSA) using high-performance liquid chromatography on stationary phases based on immobilized albumins. The effect of the addition to the mobile phase of compounds known to bind to HSA at site I (phenylbutazone) or at site II (R- and S-ibuprofen) or at both sites (2,3,5-triiodobenzoic acid) was investigated on all three proteins. The results indicated that for the chiral compounds studied, the stereoselectivity of drug binding was much lower on RatSA than on HSA. On RatSA and RabSA, the benzodiazepine site was not a major binding site for R- and S-ibuprofen. The results indicated the existence of two binding sites for R and S warfarin on RatSA and probably on RabSA. On RatSA, one site is the major stereoselective site and is the major binding site of phenylbutazone and piroxicam. The other one is a major binding site for R- and S-ibuprofen and R- and S-ketoprofen.
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