Glycans are polysaccharides formed in living organisms, in which they also play an important role. They belong to the carbohydrates. In addition to their energetic, structural and metabolic role, they form glycoconjugates with proteins and lipids, which play an important role in information transfer in organisms. Glycans are also widely used in the pharmaceutical industry for the production of drugs. Recently, the pharmaceutical industry has also increasingly relied on the principle of quality by design in the development of drugs. This leads to a systematic approach of drug development and allows for a better understanding of the process itself. As part of my master's thesis, I developed a chromatographic method for the separation and quantification of glycans on maltodextrins. Since the molecules in the sample did not contain a fluorophore, they were previously labelled with a fluorescent label, 2-aminobenzamide. Detection was performed using a fluorescence detector. Purification of excess 2-aminobenzamide was performed by solid phase extraction and liquid extraction by reaction with the organic phase. To develop the chromatographic method, 4 columns were compared, of which the Acquity Premier Glycan BEH Amide column was selected. The final chromatographic method was developed on the selected column. After development, validation of the final method was performed. The standard N-acetyl-D-glucosamine was used. After the development of the method was completed, the principle of Quality by Design was applied to the entire process. This involved optimising various parameters in each step of the process derivatization, purification and separation and checking the effect of parameter changes on sample determination.
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