Termální degradace hyaluronanu
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Vysoké učení technické v Brně. Fakulta chemická
Abstract
Tepelná stabilita a degradace hyaluronanu (HA) byla v rámci této diplomové práce zkoumána u HA s molekulovou hmotností 90–130 kDa a u HA s molekulovou hmotností 1 500–1 750 kDa. K výzkumu byly použity metody reologie, SEC-MALLS, TGA a DSC. Nízkomolekulový HA byl podroben zkoumání časové závislosti degradace, kdy byl před samotnou přípravou roztoků sušen při teplotě 90 °C po dobu 30 minut a 60 minut. U vysokomolekulového HA byla zkoumána nejen časová, ale i teplotní závislost degradace. V případě zkoumání časové závislosti degradace byl vysokomolekulový HA před přípravou roztoků sušen při teplotě 75 °C v časovém rozmezí od 15 minut do 120 minut. Při přípravě roztoků pro zjištění teplotní závislosti degradace byl pak vysokomolekulový HA sušen po dobu 30 minut v rozmezí teplot od 60 °C do 90 °C. U nízkomolekulového HA byla prokázána tepelná stabilita, nedocházelo tedy k poklesu molekulové hmotnosti a roztoky nevykazovaly výrazný pokles viskozity. U vysokomolekulového HA tepelná stabilita prokázána nebyla, k degradaci vlivem teploty sušení, ale i času sušení docházelo, což se projevilo výrazným poklesem molekulové hmotnosti a viskozity roztoků. Zatímco v případě použití teploty sušení 60 °C došlo k poklesu molekulové hmotnosti přibližně o 5 %, při teplotě sušení 90 °C se molekulová hmotnost snížila přibližně o 20 % oproti nesušenému HA. Z tohoto důvodu byl vysokomolekulový HA dále zkoumán i pomocí metod TGA, kde bylo sledováno snížení vlhkosti vzorků HA v závislosti na teplotě sušení, a DSC, kde bylo cílem zjistit teploty, při kterých dochází k odpaření vlhkosti obsažené ve vzorku HA v závislosti na jeho formě (nesušeného HA, sušeného HA a lyofilizovaného HA) a dále teplo potřebné k odpaření vlhkosti ze vzorku HA. Na závěr tohoto výzkumu byly získané výsledky u vysokomolekulového HA porovnány s výsledky jiného procesu sušení – lyofilizací, která se ukázala jako velmi šetrná metoda sušení, protože k poklesu molekulové hmotnosti u lyofilizovaného HA oproti nesušenému HA téměř nedocházelo.
This diploma thesis investigated thermal stability and the degradation of hyaluronan (HA) in HA with a molecular mass of 90–130 kDa and in HA with a molecular mass of 1 500–1 750 kDa. The following methods were used for the research: rheology, SEC-MALLS, TGA and DSC. Low-molecular HA was subject to time dependency of degradation investigation, where it was dried at a temperature of 90 °C for a period of 30 minutes and 60 minutes prior to the preparation of the solutions itself. High-molecular HA was investigated not only from the point of view of time but from the point of view of temperature dependency of degradation as well. In the case of investigating the time dependency of degradation, high-molecular HA was dried at a temperature of 75 °C at a time range from 15 minutes to 120 minutes prior to the preparation of the solutions. During the preparation of the solutions for discovering the temperature dependency of degradation, the high-molecular HA was then dried for a period of 30 minutes at a temperature range from 60 °C to 90 °C. For low-molecular HA, thermal stability was proven. Therefore, there is no decrease in the molecular mass and the solutions did not demonstrate a significant decrease of viscosity. For high-molecular HA, thermal stability was not proven. Degradation due to the temperature of drying as well as the time of drying occurred, which was demonstrated by a significant decrease in molecular mass and viscosity of the solutions. While in the case of using a drying temperature of 60 °C, a decrease in the molecular mass occurred by approximately 5 %, the molecular mass decreased by approximately 20 % at a drying temperature of 90 °C compared to undried HA. Due to this reason, high-molecular HA was also further investigated by means of the TGA method, where the decrease of humidity of HA samples in relation to the drying temperature was observed. The DSC method was also used. The objective of the DSC method was to find out temperatures, at which evaporation of humidity contained in an HA sample in relation to its form (undried HA, dried HA and lyophilized HA) occurs. This method further finds out the heat necessary to evaporate humidity from an HA sample. To conclude this research, the results obtained for high-molecular HA were compared with the results of other drying processes – lyophilized proved to be a very gentle drying method because a decrease in the molecular mass for lyophilized HA compared with undried HA almost did not occur.
This diploma thesis investigated thermal stability and the degradation of hyaluronan (HA) in HA with a molecular mass of 90–130 kDa and in HA with a molecular mass of 1 500–1 750 kDa. The following methods were used for the research: rheology, SEC-MALLS, TGA and DSC. Low-molecular HA was subject to time dependency of degradation investigation, where it was dried at a temperature of 90 °C for a period of 30 minutes and 60 minutes prior to the preparation of the solutions itself. High-molecular HA was investigated not only from the point of view of time but from the point of view of temperature dependency of degradation as well. In the case of investigating the time dependency of degradation, high-molecular HA was dried at a temperature of 75 °C at a time range from 15 minutes to 120 minutes prior to the preparation of the solutions. During the preparation of the solutions for discovering the temperature dependency of degradation, the high-molecular HA was then dried for a period of 30 minutes at a temperature range from 60 °C to 90 °C. For low-molecular HA, thermal stability was proven. Therefore, there is no decrease in the molecular mass and the solutions did not demonstrate a significant decrease of viscosity. For high-molecular HA, thermal stability was not proven. Degradation due to the temperature of drying as well as the time of drying occurred, which was demonstrated by a significant decrease in molecular mass and viscosity of the solutions. While in the case of using a drying temperature of 60 °C, a decrease in the molecular mass occurred by approximately 5 %, the molecular mass decreased by approximately 20 % at a drying temperature of 90 °C compared to undried HA. Due to this reason, high-molecular HA was also further investigated by means of the TGA method, where the decrease of humidity of HA samples in relation to the drying temperature was observed. The DSC method was also used. The objective of the DSC method was to find out temperatures, at which evaporation of humidity contained in an HA sample in relation to its form (undried HA, dried HA and lyophilized HA) occurs. This method further finds out the heat necessary to evaporate humidity from an HA sample. To conclude this research, the results obtained for high-molecular HA were compared with the results of other drying processes – lyophilized proved to be a very gentle drying method because a decrease in the molecular mass for lyophilized HA compared with undried HA almost did not occur.
Description
Citation
ŠIMÁČKOVÁ, M. Termální degradace hyaluronanu [online]. Brno: Vysoké učení technické v Brně. Fakulta chemická. 2016.
Document type
Document version
Date of access to the full text
Language of document
cs
Study field
Spotřební chemie
Comittee
prof. Ing. Miloslav Pekař, CSc. (předseda)
prof. Ing. Michal Čeppan, CSc. (místopředseda)
doc. Ing. Irena Kratochvílová, Ph.D. (člen)
doc. Ing. Marián Lehocký, Ph.D. (člen)
prof. Ing. Oldřich Zmeškal, CSc. (člen)
prof. Ing. Michal Veselý, CSc. (člen)
Date of acceptance
2016-06-01
Defence
Pekař: v jaké atmosféře byla prováděna termická analýza
Veselý: k čemu je vztažena relativní hmotnost
Result of defence
práce byla úspěšně obhájena
Document licence
Standardní licenční smlouva - přístup k plnému textu bez omezení