Plasma protein binding study of oxybutynin by high-performance frontal analysis

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2002-02, Vol.768 (1), p.177-188
• Main Authors: Shibukawa, Akimasa, Ishizawa, Nobuko, Kimura, Tomoko, Sakamoto, Yuki, Ogita, Kanae, Matsuo, Yuka, Kuroda, Yukihiro, Matayatsuk, Chutima, Nakagawa, Terumichi, Wainer, Irving W
• Format: Article
• Language: English
• Subjects: Blood Proteins - metabolism; Chromatography, High Pressure Liquid - methods; Humans; Mandelic Acids - metabolism; Oxybutynin; Protein Binding
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/S0378-4347(01)00497-2

Plasma protein binding of oxybutynin (OXY) was investigated quantitatively and enantioselectively using high-performance frontal analysis (HPFA). An on-line HPLC system which consists of HPFA column, extraction column and analytical column was developed to determine the unbound concentrations of OXY enantiomers in human plasma, in human serum albumin (HSA) solutions, and in human α 1-acid glycoprotein (AGP) solutions. OXY is bound in human plasma strongly and enantioselectively. The bound drug fraction in human plasma containing 2–10 μ M ( R)- or ( S)-OXY was higher than 99%, and the unbound fraction of ( R)-OXY was 1.56 times higher than that of ( S)-isomer. AGP plays the dominant role in this strong and enantioselective plasma protein binding. The total binding affinities ( nK) of ( R)- and ( S)-OXY to AGP were 6.86×10 6 and 1.53×10 7 M −1, respectively, while the nK values of ( R)- and ( S)-OXY to HSA were 2.64×10 4 and 2.19×10 4 M −1, respectively. The binding affinity of OXY to AGP is much higher than that to HSA, and shows high enantioselectivity ( S/ R ratio of nK values is 2.2). It was found that both enantiomers are bound competitively at the same binding site on an AGP molecule. The binding property between OXY and low density lipoprotein (LDL) was investigated by using the frontal analysis method incorporated in high-performance capillary electrophoresis (HPCE/FA). It was found the binding is non-saturable and non-enantioselective.