Premium
Enantioseparation of nuarimol by affinity electrokinetic chromatography‐partial filling technique using human serum albumin as chiral selector
Author(s) -
MartínezGómez Maria Amparo,
EscuderGilabert Laura,
VillanuevaCamañas Rosa M.,
Sagrado Salvador,
MedinaHernández Maria J.
Publication year - 2008
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200800190
Subject(s) - enantiomer , human serum albumin , chromatography , electrokinetic phenomena , chemistry , resolution (logic) , electrophoresis , analytical chemistry (journal) , capillary electrophoresis , stereochemistry , artificial intelligence , computer science
The present paper deals with the enantiomeric separation of nuarimol enantiomers by affinity EKC‐partial filling technique using HSA as chiral selector. Firstly, a study of nuarimol interactions with HSA by CE‐frontal analysis was performed. The binding parameters obtained for the first site of interaction were n 1 = 0.84; K 1 = 9.7 ± 0.3×10 3 M –1 and the protein binding percentage of nuarimol at physiological concentration of HSA was 75.2 ± 0.2%. Due to the moderate affinity of nuarimol towards HSA the possibility of using this protein as chiral selector for the separation of nuarimol using the partial filling technique was evaluated. A multivariate optimization approach of the most critical experimental variables in enantioresolution, running pH, HSA concentration and plug length was carried out. Separation of nuarimol enantiomers was obtained under the following selected conditions: electrophoretic buffer composed of 50 mM Tris at pH 7.3; 160 μM HSA solution applied at 50 mbar for 156 s as chiral selector; nuarimol solutions in the range of 2–8×10 –4 M injected hydrodynamically at 30 mbar for 2 s and the electrophoretic runs performed at 30°C applying 15 kV voltage. Resolution, accuracy, reproducibility speed and cost of the proposed method make it suitable for quality control of the enantiomeric composition of nuarimol in formulations and for further toxicological studies. The results showed a different affinity between nuarimol enantiomers towards HSA.