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Analysis of vancomycin and related impurities by micellar electrokinetic capillary chromatography. Method development and validation
Author(s) -
Kang Jingwu,
Van Schepdael Ann,
Roets Eugène,
Hoogmartens Jos
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200107)22:12<2588::aid-elps2588>3.0.co;2-x
Subject(s) - detection limit , chromatography , micellar electrokinetic chromatography , repeatability , capillary electrophoresis , chemistry , tris , capillary action , selectivity , buffer solution , electrokinetic phenomena , central composite design , analytical chemistry (journal) , response surface methodology , materials science , biochemistry , composite material , catalysis
A fast and highly selective micellar electrokinetic capillary chromatography (MEKC) method for quantitative analysis of vancomycin and related impurities is described. Among the tested surfactants, cetyltrimethylammonium chloride (CTAC) offered the best selectivity. Another important parameter, which strongly influenced the selectivity, was buffer pH. It was found that the selectivity increased with buffer pH decreasing from 9 to 5. Using Tris‐phosphate buffer containing CTAC, satisfactory separation could be obtained in the pH range from 5.0 to 5.5. Excellent repeatability in terms of migration time and peak area could be obtained when the capillary was carefully washed between two runs. In order to obtain optimal conditions and to evaluate the method robustness, a central composite experimental design was carried out. The optimal conditions were: 44 cm length of fused‐silica capillary with 50 νm ID, 120 m M Tris‐phosphate buffer (pH 5.2) containing 50 m M CTAC, –15 kV applied voltage, UV detection at 210 nm, and a column temperature of 25°C. Under the optimal conditions, more than 20 peaks could be separated within 8 min. The method has a linearity range from 0.004 to 1.2 mg/ml (concentration of vancomycin B, active component). The limit of detection (LOD) and limit of quantitation (LOQ) were 0.4 νg/mL vancomycin, equivalent to 0.3 νg/mL vancomycin B (0.04%) and 1.1 νg/mL vancomycin, equivalent to 0.9 νg/mL vancomycin B (0.1%), respectively.