Abstract
Capillary zone electrophoresis was successfully applied to separate eight related substances of kanamycin and several minor unknowns from the main component. Strategies to enhance derivatization and selectivity and to optimize separation parameters involved the application of experimental designs. This chemometrical approach considers main effects as well as interactions of the influential parameters, thus conducting a more thorough investigation of the method than the common step-by-step approach. Central composite face centered designs established optimal separation conditions: 30 m M borax buffer, pH 10.0 containing 16.0% (v/v) methanol and optimal composition of derivatization reagent: 27 mg/ml 1,2-phthalic dicarboxaldehyde and 25 μl/ml mercaptoacetic acid in borate buffer, pH 10.4. The standard curves were linear over the concentration range of 0.007–1.01 mg/ml for the main component and 0.003–0.1 mg/ml for the related substances. The limit of quantitation was 0.14% (m/m) for the related substances and impurities ( S/ N=10). The assay method was used to determine the composition of several commercial samples. Quantitative analysis indicates potential usefulness of capillary electrophoresis as an alternative to the assay method prescribed in the European Pharmacopoeia and the United States Pharmacopeia.
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