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Separation and assay of indomethacin and its degradation products by micellar electrokinetic chromatography
Mitlenerová, Dagmar ; Michalíková, Klára (advisor) ; Honegr, Jan (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Dagmar Mitlenerová Supervisor: PharmDr. Klára Petrů, Ph.D. Consultant: doc. RNDr. Miroslav Polášek, CSc. Title of Diploma Thesis: Micellar electrokinetic chromatography for the separation and assay of indomethacin and its degradations products A new method of micellar electrokinetic chromatography (MEKC) for analysis of indomethacin and its degradation products, 4-chlorbenzoic acid (4CHBA) and 5-methoxy- 2methy-3indolylacetic acid (MMIAA), was developed and optimized. The separation was carried out in uncoated fused silica capillary (with i.d. 75 µm, total length 33 cm and effective length 24,5 cm) with applied voltage 15 kV and temperature of capillary 25ř C. Wave length for UV detection was 224 nm and 234 nm. The optimized background electrolyte contained: 10mM phosphate buffer, 60mM SDS (pH 7,0) and 10% (v/v) of methanol. Internal standard was 1-naftylacetic acid. Large volume sample stacking method was used for sensitivity enhancement. The mixture of standards was injected hydrodynamically at 100 mBar for 10 s (24% of capillary total length). Voltage -7,5 kV was applied for stacking of the analytes. Polarity was switched to +15 kV (time of switching 1,5 min), when 95% of original...
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