20.500.12556/RUL-169885 Priprava vzorcev svile za velikostno izključitveno kromatografijo Silk sample preparation for size exclusion chromatography Svila je že stoletja cenjen material. Za zaščito zgodovinsko pomembnih izdelkov narejenih iz svile, je potrebno ugotoviti stopnjo njene razgradnje. To lahko ugotovimo s spremljanjem spremembe dolžine verig proteina fibroina, ki je glavni sestavni del svile. Porazdelitev molske mase fibroina lahko preučujemo z uporabo velikostno izključitvene kromatografije (SEC). Optimizirala sem način priprave vzorcev svile za SEC analizo in analizirala različne vrste svile. Analizirala sem svile iz treh različnih virov (slovensko, indijsko in belgijsko), ki so bile obdelane na različne načine: neobdelana, različno obtežene svile, svilo pospešeno razgrajeno na štiri različne načine in svilo, ki sem ji dodala tripsin. Izkazalo se je, da je optimalna priprava vzorcev sledeča: vzorce sem raztapljala v koncentriranem LiBr, nato pa jih 6 h dializirala, 1 h v MQ-vodi in 5 h v citratnem pufru uporabljenim za mobilno fazo, na koncu pa sem jih centrifugirala. Raziskala sem stabilnost vzorca v LiBr in ugotovila, da vzorec s časom v LiBr razpada. Ugotovila sem, da različni načini pospešene razgradnje svile različno vplivajo na njeno razgradnjo, predvsem jo pospešujeta višja temperatura in večja vlažnost. Obteževanje svile je otežilo njeno raztapljanje, saj se obtežena svila z izbranim postopkom ni povsem raztopila, zato sem z metodo FTIR preverila, kateri del svile ostane neraztopljen in ugotovila, da neraztopljeni del vsebuje tudi fibroin. For centuries, silk has been a highly valued material. To protect historic silk materials, it is necessary to assess the degree of its degradation. This can be observed through the changes in chain lenght of protein fibroin, which is the main component of silk. The molar mass distribution of fibroin can be determined using size exclusion chromatography (SEC). I optimized the silk sample preparation method for SEC analysis and I analyzed different types of silk. I analyzed silks of three different origins (Slovenian, Indian and Belgian), different weighted silks, four different degraded silks and silks treated with trypsin. It turned out that the optimum sample preparation is as follows: samples were dissolved in concentrated LiBr, then dialyzed for 6 h, 1 h in MQ-water and 5 h in the citrate buffer used for the mobile phase and centrifuged at the end. Stability of the samples in LiBr has been investigated and found that the sample decays with time in LiBr. I found that different methods of accelerated degradation of silk had different effects on its degradation, with higher temperature and higher humidity accelerating the process. Silk weighting made its dissolution more difficult, as the weighted silk did not completely dissolve using the selected process. FTIR method analysis was conducted to determine which components of silk remain undissolved and it was revealed that part of the fibroin remained undissolved. svila fibroin SEC razgradnja silk fibroin SEC degradation true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2025-06-13 13:05:05 2025-06-13 13:05:30 2025-06-18 08:25:35 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 25363 239750403 Lipoglavsek_Kaja_-_Priprava_vzorcev_svile_za_velikostno_izkljucitveno_kromatografijo.pdf Lipoglavsek_Kaja_-_Priprava_vzorcev_svile_za_velikostno_izkljucitveno_kromatografijo.pdf 1 9757930A85C609A8B92D932F3E6C4615 0a6d5fa8aff134ef72fe78aada1a0f9137a86a6d0302ba0a8e53b9723bfae675 cb6c3615-4844-11f0-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=211646 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0