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Development and validation of a robust capillary electrophoresis method for impurity profiling of calcium levofolinate including the (6 R ,2' S )‐diastereomer using statistical experimental design
Author(s) -
Süß Falko,
Harang Valérie,
Sängervan de Griend Cari E.,
Scriba Gerhard K. E.
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200305730
Subject(s) - capillary electrophoresis , chromatography , chemistry , impurity , detection limit , linearity , factorial experiment , analytical chemistry (journal) , diastereomer , linear regression , electrolyte , robustness testing , accuracy and precision , mathematics , stereochemistry , electrode , statistics , computer science , programming language , physics , organic chemistry , quantum mechanics , software
A chiral capillary electrophoresis assay for the simultaneous determination of the optical purity and of related substances of calcium levofolinate has been developed and validated. Using 2,6‐dimethyl‐β‐cyclodextrin as chiral selector at a concentration of 20 mg/mL, the method was optimized using a full factorial design with four factors including pH and concentration of the background electrolyte, column temperature and separation voltage. Optimized conditions were a 40 m M sodium tetraborate buffer, pH 9.9, a capillary temperature of 16°C, and an applied voltage of 21 kV. Methotrexate was used as internal standard to compensate for injection errors and fluctuations of the migration times. A multiple linear regression model was also used to test the robustness of the method. Validation was performed with respect to specificity, linearity, range, limit of quantification and detection, precision, and accuracy. The assay allowed the detection and determination of related substances including the diastereomeric (6 R ,2' S )‐impurity of levofolinic acid at the 0.1% level, the identification threshold of impurities for orally administered drugs for human use defined by the International Conference on Harmonization guidelines as well as the European Pharmacopoeia.