Table of Contents Author Guidelines Submit a Manuscript
Journal of Analytical Methods in Chemistry
Volume 2013 (2013), Article ID 439039, 7 pages
Research Article

Rapid Screening of Drug-Protein Binding Using High-Performance Affinity Chromatography with Columns Containing Immobilized Human Serum Albumin

1Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China
2Institute of Pharmacy, Medical School of Xiang Ya, Central South University, Changsha 410013, China
3Hepatobiliary Surgery Department, The Affiliated Hospital of Medical College, Qingdao University, Qingdao 266003, China
4Department of Pharmacy, Lushan Sanitarium of PLA, Jiujiang 332000, China

Received 25 January 2013; Accepted 28 February 2013

Academic Editor: Shuang-Qing Zhang

Copyright © 2013 Ying-Fei Li et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


For drug candidates, a plasma protein binding (PPB) more than 90% is more meaningful and deserves further investigation in development. In the study, a high-performance liquid chromatography method employing column containing immobilized human serum albumin (HSA) to screen in vitro PPB of leading compounds was established and successfully applied to tested compounds. Good correlation (a coefficient correlation of 0.96) was attained between the reciprocal values ( ) of experimentally obtained retention time of reference compounds eluted through HSA column and the reported PPB values ( ) with a correlation equation of . The method was successfully applied to six test compounds, and the result was confirmed by the conventional ultrafiltration technique, and both yielded equal results. However, due to the particular protein immobilized to column, the method cannot be applied for all compounds and should be exploited judiciously based on the value of the logarithmic measure of the acid dissociation constant (pKa) as per the requirement. If 1-acid glycoprotein and other plasma proteins could be immobilized like HSA with their actual ratio in plasma to column simultaneously, the result attained using immobilized column may be more accurate, and the method could be applied to more compounds without pKa limitation.