Analysis of polyanionic macromolecular carrier poly-(N-vinylpyrrolidone-co-maleic acid) and its bioconjugats by capillary electrophoresis.

Abstract

Anionic carrier poly(N-vinylpyrrolidone-co-maleic acid) and its conjugates, prepared with coupling of 2-cyano-3-hydroxy-5-amino-2-pentenoyc(4-trifluoromethyl anilide) or (6', 7'-dimethyl-l'-quinoxalinyl)-4-(2' amino) acetanilide to the carrier, were analyzed by capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) in the following buffers: 0.25 N triethylammonium phosphate (TEAP); sodium dodecyl sulfate (SDS; 25-150 mM) in TEAP (pH 2.25-6.30); 0.1 N Na-borate buffer (pH range 7-11) and SDS (25-150 mM) in Na-borate buffer (pH range 7-11). The presence of strong carboxyl groups (dissociated even at pH 2.25) on the polymer chain was proved by the CZE method. It was also proved by potentiometric titration that carboxyls with a wide range of acidity were on the polymer chain. CZE was able to differentiate among the analytes possessing carboxyl groups of different acidic strengths at pH 2.25. These components were not distinguished by CZE at high pH values (11.0). Interaction between the analyte and SDS affected the separation at this pH, and hence good resolution was obtained by MEKC. Informative separations were achieved both for the carrier and the conjugates in TEAP buffer at pH 2.25 by the CZE method. Optimal separation was achieved in borate buffer containing 75 mM SDS at pH 11.0 for the carrier and at pH 7.7 for the conjugates in MEKC.