Evaluation of quail egg white riboflavin binding protein as a chiral selector in capillary electrophoresis by applying a modified partial filling technique.
A preliminary evaluation of the enantioselective properties of quail egg yolk riboflavin binding protein (qRfBP) was carried out in capillary electrophoresis by using the complete filling technique. The most promising results obtained by this screening of nineteen chiral drugs were singled out with the aim of optimizing enantiomer separations by applying the partial filling technique, which allows operating at much higher protein concentrations without detection problems. The building of the separation zone in the partial filling technique has been modified in order to enable on-line monitoring, before each run, of the actual protein plug application velocity and, consequently, the building of a plug of the desired length. The electrophoretic conditions chosen gave opposite migration directions for the chiral selector and the analytes, with qRfBP migrating away from the detector. A polyvinyl alcohol-coated capillary was first totally filled with protein and the optimal plug length was obtained by further applying negative pressure together with positive voltage for the time needed. Separations of basic drugs were optimized by using protein concentrations ranging from 200 microM up to 900 microM and different plug lengths, while the running buffer pH (6.0), temperature (25 degrees C) and operating voltage (+20 kV) were kept constant. The enantioresolution of all solutes was affected by both the chiral selector concentration and protein plug length. Baseline separations were obtained for oxprenolol, prilocaine and bupivacaine.
De Lorenzi
E
E
Department of Pharmaceutical Chemistry, University of Pavia, Italy. ersilia@chifar.unipv.it
Massolini
G
G
Quaglia
M
M
Galbusera
C
C
Caccialanza
G
G
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry*; D004529:Egg White / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005069:Evaluation Studies as Topic; D026901:Membrane Transport Proteins; D011784:Quail; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism
D002352:Carrier Proteins; D026901:Membrane Transport Proteins; C019079:riboflavin-binding protein