magistrsko delo
Povzetek
V magistrski nalogi je predstavljen razvoj tekočinske kromatografske metode, ki temelji na tehniki ločevanja z reverzno fazo. Metoda je bila razvita za potrebe določanja aminokislin v formulatorskih vzorcih bioloških zdravil. Z uporabo instrumentacije HPLC, kolone Acclaim RSLC Polar Advantage II in detektorja CAD smo razvili metodo za določanje 13 aminokislin, ki so najpogosteje prisotne v formulatorskih vzorcih. S številnimi analitskimi postopki razvoja in optimizacije metode nam je uspelo razviti točno, robustno in hkrati preprosto kromatografsko metodo, pri kateri za ločbo uporabljamo kromatografsko kolono RP polar embeded. Ključni parametri razvoja metode so bili določitev sestave polarne in nepolarne mobilne faze, postavitev gradienta metode s spreminjanjem deleža mobilnih faz, temperatura kolone, temperatura detektorja in volumen injiciranja vzorca. Za določanje aminokislin smo uporabili Corona Veo RS detector (CAD), saj detektorji na podlagi fluorescence niso bili primerni, zaradi pomanjkanja fotokromov aminokislin. Poleg razvoja metode pa je delo v laboratoriju obsegalo tudi pripravo vzorcev in standardov, pripravo mobilnih faz in najpomembnejše – pripravo in ekvilibracijo instrumenta HPLC. Kot rezultat dela smo razvili kromatografsko metodo za določanje aminokislin v vzorcih bioloških zdravil.
Ključne besede
kromatografija;aminokisline;kromatografske metode;modifikatorji;razvoj;
Podatki
Jezik: |
Slovenski jezik |
Leto izida: |
2020 |
Tipologija: |
2.09 - Magistrsko delo |
Organizacija: |
UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo |
Založnik: |
[T. Škerlavaj] |
UDK: |
606:615:604.2:661.745:543.544.5HPLC(043.2) |
COBISS: |
43966467
|
Št. ogledov: |
458 |
Št. prenosov: |
94 |
Ocena: |
0 (0 glasov) |
Metapodatki: |
|
Ostali podatki
Sekundarni jezik: |
Angleški jezik |
Sekundarni naslov: |
Development of chromatographic method for analysis of selected amino acids in biological medicines |
Sekundarni povzetek: |
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. |
Sekundarne ključne besede: |
chromatography;amino acids;chromatographic methods;modifiers;development; |
Vrsta dela (COBISS): |
Magistrsko delo/naloga |
Študijski program: |
0 |
Komentar na gradivo: |
Univ. v Ljubljani, Biotehniška fak., Študij biotehnologije |
Strani: |
X, 67 f. |
ID: |
12322826 |