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Utilization of ultra-high performance liquid chromatography for simultaneous determination of paracetamol, amoxicillin, and doxycycline
Hrašková, Hana ; Kubíčková, Anna (advisor) ; Sobotníková, Jana (referee)
In this study, a method was developed for the simultaneous determination of three pharmaceutical compounds: amoxicillin, doxycycline, and paracetamol. Ultra-high performance liquid chromatography (UHPLC) was employed for the separation. A Kinetex column C18 measuring 100×2,1 mm with a particle size of 1,7 μm was used as the stationary phase. The optimal method was run in gradient mode with a total analysis time of 5 minutes. The mobile phase consisted of a mixture of component A, containing acetonitrile with 0,1% formic acid, and component B, comprising deionized water with 0,1% formic acid. The separation conditions in the column varied according to the following gradient program: 0 - 1,0 min: 10% A (v/v); 1,0 - 1,5 min: 10% → 65% A (v/v); 1,5 - 2,0 min: 65% A (v/v); 2,0 - 2,5 min 65%→10% A (v/v); 2,5 - 5,0 min 10% A (v/v). The pH of the mobile phase was maintained at 2,8. The flow rate was set at 0,3 ml/min, column temperature was maintained at 25řC, and the autosampler temperature was consistently held at 8řC. The injection volume was 1 μl, and analytes were detected using a diode array detector at a wavelength of 245 nm. Drug identification was conducted using a QDA detector set to positive mode with a mass range from 100,00 to 500,00 Da. The method was partially validated by constructing a...
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