Separation of enantiomers on chiral stationary phase based on cellulase: Effect of preparation method and silica particle diameters on chiral recognition ability.
Cellulase (Cel) was immobilized onto aminopropyl-silica gels via its amino and carboxy groups, respectively, using N,N'-disuccinimidyl carbonate, and 1-ethyl-3-(3'-dimethylaminopropyl)carbodimide and N-hydroxysulfosuccinimide. They were termed N-Cel and C-Cel, respectively. Despite their smaller retention factors on a C-Cel column, the enantioseparation factors and resolution of β-blockers, propranolol, alprenolol, oxprenolol and pindolol, were similar with N- and C-Cel columns. In addition, C-Cel was prepared using aminopropyl-silica gels, whose nominal particle diameters were 5 and 3, and 2.1μm, respectively. A C-Cel column prepared with 2.1-μm aminopropyl-silica gels gave the highest enantioselectivity and column efficiency among three C-Cel columns. Furthermore, the influence of N,N-dimethyl-n-octylamine (DMOA) or cellobiose concentrations on the retentivity and enantioselectivity for β-blockers on a C-Cel column was investigated. The results indicate that single-site competition of β-blockers with DMOA or cellobiose on the catalytic binding site of Cel and the further bindings at the secondary site in a non-competitive fashion could occur. Furthermore, the enantioselective bindings of β-blockers could occur at the catalytic biding cite of Cel and at the secondary binding site.
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University, 11-68, Koshien Kyuban-cho, Nishinomiya 663-8179, Japan.
Hisami
H
Matsunaga
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University, 11-68, Koshien Kyuban-cho, Nishinomiya 663-8179, Japan. Electronic address: haginaka@mukogawa-u.ac.jp.
Jun
J
Haginaka
D000319:Adrenergic beta-Antagonists / Q000032:analysis / Q000737:chemistry; D001665:Binding Sites; D002384:Catalysis; D002480:Cellulase / Q000737:chemistry*; D002623:Chemistry Techniques, Analytical / Q000379:methods* / Q000592:standards; D010316:Particle Size; D012822:Silicon Dioxide / Q000737:chemistry*; D013237:Stereoisomerism
D000319:Adrenergic beta-Antagonists; C021611:aminopropyl silica gel; D012822:Silicon Dioxide; D002480:Cellulase
Alprenolol; Cellulase; Chiral stationary phase; Enantiomer separation; Propranolol