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Development of an ultra-high performance liquid chromatography for rifampicin determination
Alušicová, Sára ; Kozlík, Petr (advisor) ; Hraníček, Jakub (referee)
In this bachelor thesis, an ultra-high-performance liquid chromatography method (UHPLC) was developed to determine rifampicin in phosphate buffer (also called PBS). Rifampicin is a bactericidal antibiotic from the rifamycin family, which plays an important role in the treatment of tuberculosis. Optimized conditions for rifampicin determination in PBS were as follows: column kinetex UHPLC C18 100 (50x2.1 mm; 1.3 µm), mobile phase consisted of 10 mM aqueous ammonium formate buffer (pH 3.51) to which trifluoroacetic acid was added as an ion pairing agent (to a final concentration of 0.1%) (component A) and of ACN (component B). The measurement was carried out in the gradient elution mode of the mobile phase (time: 0-1-6-7-8-11, component B: 30,30,80,80,30,30). The flow rate was 0.300 ml / min at a column temperature of 30 ř C. The injection volume was 5µl. Under these optimized UHPLC conditions, a calibration dependence was measured in concenatraction range from 28,310-3 mg ml-1 to 4,4210-4 mg ml-1 . The limit of detection was 1,05∗10-4 mg ml -1 and the limit of quantification was 3,5∗10-4 mg ml -1 . Keywords: antibiotic, rifampicin, UHPLC
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