Separacija glikanov s kromatografijo z mešanimi režimi

Žohar, Jona (author), Pompe, Matevž (mentor)

Glikani so ogljikovi hidrati, ki imajo pomembno strukturno in metabolično vlogo v organizmih, zaradi česar se pogosto uporabljajo tudi v farmacevtski industriji. V diplomskem delu, sem na maltodextrinih razvila metodo za separacijo glikanov s kromatografijo z mešanimi režimi. Vzorec maltodextrinov, na katerem sem razvila metodo, sem označila s fluorescenčnim označevalcem in odvečni označevalec odstranila z ekstrakcijo na trdno fazo. Metodo sem razvila na Thermo Fisher-jevi koloni, GlycanPac AXH-1, na kateri poteka več mehanizmov separacije, in sicer ionske in hidrofilne interakcije. Razvito metodo sem nato validirala z D-glukozo in D-maltozo. Pri čemer smo preverjali linearnost, občutljivost, ponovljivost, točnost in robustnost.

• Language: Slovenian
• Keywords: Glikani, Ogljikovi hidrati, 2-AA, SPE, Kromatografija z mešanimi režimi
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2020
• PID: 20.500.12556/RUL-119650
• COBISS.SI-ID: 30501891
• Publication date in RUL: 10.09.2020

Secondary language

• Language: English
• Title: Separation of glycans using mixed-mode chromatography

Abstract

Glycans are carbohydrates that have an important structural and metabolic role in living organisms, which makes them commonly used in the pharmaceutical industry. As a part of my bachelor's thesis, I developed a method for the separation of glycans using mixed mode chromatography. The method was developed on maltodextrines, which were first labelled with fluorescence 2-AA reagent. I removed the excess of the fluorescence reagent using a solid phase extraction. The method was developed on the Thermo Fisher’s GlycanPac AXH-1 column, which has two mechanisms of separation, namely the ion-exchange and hydrophilic interactions. After its development, the method was validated with D-glucose and D-maltose. In order to validate the method, I checked its linearity, repeatability, sensitivity, accuracy and robustness.

• Keywords: Glycans, Carbohydrates, 2-AA, SPE, Mixed-mode chromatography