The master's thesis presents the development of a liquid chromatographic method, which is based on the technique of reverse phase separation. The method was developed for determining amino acids in formulator samples of biological drugs. Using HPLC instrumentation, an Acclaim RSLC Polar Advantage II column and a CAD detector, we developed a method for determining the thirteen most commonly present amino acids in formulation samples. Through a series of analytical procedures for the development and optimization of the method, we were able to develop an accurate, robust and at the same time simple chromatographic method in which we use the RP polar embeded chromatographic column as a separation. The key parameters of the method development were to determine the composition of the polar and non-polar mobile phase, to set the gradient of the method, which also include changing the proportion of polar and non-polar mobile phases, column temperature, detector temperature and sample injection volume. We used a Corona Veo RS detector (CAD) to determine amino acids because fluorescence-based detectors were not suitable due to a lack of amino acid photo chromes. In addition to the development of the method, the work in the laboratory also included the preparation of samples and standards, the preparation of mobile phases and, most importantly, the preparation and equilibration of the HPLC instrument. As a result of the experimental work, we developed a chromatographic method for the determination of amino acids in samples of biological drugs.
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