Micellar electrokinetic chromatography with short-end injection for rapid separation and simultaneous determination of aesculin, aesculetin, and phenylephrine in pharmaceutical preparations

• Published in: Electrophoresis Vol. 36; no. 21-22; pp. 2754 - 2761
• Main Authors: Pincova, Lucie, Petr, Klara, Pospisilova, Marie, Polasek, Miroslav
• Format: Journal Article
• Language: English
• Published: Germany Blackwell Publishing Ltd 01.11.2015
• Subjects: Aesculetin; Aesculin; Analytical chemistry; Assaying; Chromatography; Electrokinetics; Electrophoresis; Methyl alcohol; Micellar electrokinetic chromatography; Pharmaceuticals; Phenylephrine; Separation; Short-end injection; Aesculin; Aesculetin; Short-end injection; Micellar electrokinetic chromatography; Phenylephrine
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.201500215

A fast micellar electrokinetic chromatography (MEKC) method for simultaneous assay of aesculin, aesculetin, and phenylephrine was developed and validated. The separation was carried out in a fused‐silica capillary (50 μm id, total length 64.5 cm, effective length 8.5 cm) with UV detection at 210 nm, temperature 25°C and separation voltage –25 kV. The samples were loaded hydrodynamically at a pressure of –50 mbar for 6 s. The background electrolyte of pH 8.6 contained 20 mM boric acid, 60 mM SDS, and 5% (v/v) of methanol. The calibration curves were linear in the range 10−500 μg/mL for aesculin and aesculetin and 12.5–625 μg/mL for phenylephrine. The RSD values of corrected peak areas were 0.6−1.2% (n = 6) when determining 0.2 mg/mL of aesculin and aesculetin and 0.25 mg/mL of phenylephrine in prepared standard mixtures. The method was successfully applied to the assay of aesculin and phenylephrine in a pharmaceutical preparation (RSD  =  1.9−2.0%; n  =  3) and the robustness of the method for both, the determination of analytes and the system suitability test parameter values, was evaluated with the use of Plackett‐Burman design.