magistrsko delo
Jona Žohar (Author), Matevž Pompe (Mentor), Romana Cerc Korošec (Thesis defence commission member), Drago Kočar (Thesis defence commission member)

Abstract

Glikani so polisaharidi, ki nastanejo v živih organizmih, v katerih imajo tudi pomembno vlogo. Spadajo med ogljikove hidrate. Poleg njihove energijske, strukturne in metabolične vloge skupaj s proteini in lipidi tvorijo glikokonjugate, ti pa imajo pomembno vlogo pri prenašanju informacij v organizmih. Glikane veliko uporabljajo tudi v farmacevtski industriji pri proizvodnji zdravil. Zadnje čase se farmacevtska industrija vedno bolj poslužuje tudi principa upravljanja kvalitete pri razvoju zdravil. Ta prinaša sistematični pristop k razvoju zdravil in s tem omogoča večje razumevanje samega procesa. V sklopu magistrskega dela sem na maltodekstrinih razvila kromatografsko metodo za separacijo in kvantifikacijo glikanov. Zaradi pomankanja fluorofora v molekulah vzorca sem te predhodno označila s fluorescenčnim označevalcem, 2-aminobenzamidom, saj je detekcija potekala s fluorescenčnim detektorjem. Čiščenje odvečnega 2 aminobenzamida sem izvedla z ekstrakcijo na trdno fazo ter s tekočinsko ekstrakcijo preko reakcije z organsko fazo. Za razvoj kromatografske metode sem med seboj primerjala 4 kolone, izmed katerih sem izbrala kolono Acquity Premier Glycan BEH Amide, ter na njej razvila končno kromatografsko metodo. To sem validirala s standardom N-acetil-D-glukozamin. Po končanem razvoju metode sem izvedla princip upravljanja kvalitete na celotnem postopku. S tem sem v vsakem koraku postopka - derivatizaciji, čiščenju in separaciji - optimizirala različne parametre in preverila vpliv sprememb parametrov na določitev vzorca.

Keywords

2-aminobenzamid;ekstrakcija;separacija glikanov;HPLC;detekcija glikanov;princip upravljanja kvalitete;validacija kromatografske metode;magistrska dela;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher: [J. Žohar]
UDC: 543.544.5HPLC(043.2)
COBISS: 146303235 Link will open in a new window
Views: 11
Downloads: 1
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Other data

Secondary language: English
Secondary title: Application of the quality by design principle in the development of an analytical procedure for the determination of glycans
Secondary abstract: 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.
Secondary keywords: glycans;2-aminobenzamide;extraction;HPLC;quality by design;Polisaharidi;Tekočinska kromatografija;Univerzitetna in visokošolska dela;
Type (COBISS): Master's thesis/paper
Study programme: 1000375
Thesis comment: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, smer Kemija
Pages: 66 str.
ID: 18347628