Validation of a Rapid Equilibrium Dialysis Approach for the Measurement of Plasma Protein Binding

• Published in: Journal of pharmaceutical sciences Vol. 97; no. 10; pp. 4586 - 4595
• Main Authors: Waters, Nigel J., Jones, Rachel, Williams, Gareth, Sohal, Bindi
• Format: Journal Article
• Language: English
• Published: Hoboken Elsevier Inc 01.10.2008; Wiley Subscription Services, Inc., A Wiley Company; Wiley; American Pharmaceutical Association
• Subjects: albumin; alpha 1-acid glycoprotein; Biological and medical sciences; Blood Proteins - metabolism; Chromatography, Liquid; Dialysis; equilibrium dialysis; General pharmacology; high-throughput screening; Humans; Medical sciences; Pharmaceutical technology. Pharmaceutical industry; pharmacokinetics; Pharmacology. Drug treatments; Protein Binding; rapid equilibrium dialysis; Tandem Mass Spectrometry; albumin; alpha 1-acid glycoprotein; high-throughput screening; pharmacokinetics; equilibrium dialysis; rapid equilibrium dialysis; protein binding; High throughput screening; Measurement; Validation; Biological fluid; Pharmaceutical technology; Glycoprotein; Albumin; Blood plasma; Binding protein; Acids; Dialysis; Pharmacokinetics; Quantitative analysis
• ISSN: 0022-3549; 1520-6017; 1520-6017
• DOI: 10.1002/jps.21317

Equilibrium dialysis (ED) is one of the most frequently used approaches to investigate drug binding, where the major drawbacks are the time to reach equilibrium (varying between 6 and 24 h), a long assay preparation time and complexity of automation. A rapid equilibrium dialysis (RED) device has recently become commercially available (Pierce Biotechnology, ThermoFisher Scientific, Waltham, MA) offering the potential for reduced preparation and equilibration times. The RED device comprises a Teflon base plate which holds up to 48 disposable dialysis cells. Each dialysis insert is made up of two side-by-side chambers separated by a vertical cylinder of dialysis membrane with a high membrane surface area-to-volume ratio. An independent validation of the RED approach for the measurement of human plasma protein binding (PPB) was carried out as a comparative analysis with standard ED evaluating equilibration time, assay reproducibility and accuracy and ease of use. Using a diverse set of 18 commercially available drugs spanning a range of physicochemical properties we have shown this to be a robust and accurate methodology, with a shorter preparation and dialysis time, capable of being automated as a high-throughput assay for the determination of PPB. © 2008 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 97:4586–4595, 2008