Charakterizace koloidních částic pomocí deprotonace v excitovaném stavu za použití pokročilých fluorescenčních technik
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Vysoké učení technické v Brně. Fakulta chemická
Abstract
V této diplomové práci byly studovány deprotonační charakteristiky fluorescenčních sond -naftolu a 8-hydroxypyren-1,3,6-trisulfonové kyseliny (HPTS), pomocí měření ustálené a časově rozlišené fluorescence. Byly měřeny dva kationtové tenzidy, Septonex a cetyltrimethylamonium bromid (CTAB) v komplexu s hyaluronanem o třech molekulových hmotnostech, 1,75 MDa, 1 MDa a 300 kDa. Měřením ustálené fluorescence byly stanoveny kritické agregační koncentrace pro jednotlivé tenzidy a také hodnoty pKa*. Dále byly měřením časově rozlišené fluorescence stanoveny deprotonační konstanty a průměrné doby života. Ze získaných dat a jejich porovnáním se systémem polystyrensulfonát (PSS) – tenzid byl posuzován vliv hydratačního obalu hyaluronanu na vznik komplexu hyaluronan – tenzid. Byly zjištěny velké rozdíly v deprotonačních charakteristikách mezi jednotlivými tenzidy a mezi jednotlivými molekulovými hmotnostmi hyaluronanu. Z měření plyne lokalizace hydratačního obalu v okolí disociované karboxylové skupiny, kde probíhá interakce s kladným nábojem tenzidu za vzniku komplexu. Dále bylo pomocí zhášení pyrenu cetylpyridinium chloridem (CPC) stanoveno agregační číslo Septonexu pro koncentraci 20mM na hodnotu 104 molekul. Pomocí CPC byla také potvrzena lokalizace -naftolu v micelách CTAB, respektive v komplexu polymer – CTAB.
The deprotonation characteristics of fluorescent probes -naphthol and 8-hydroxypyrene-1,3,6-trisulphonic acid (HPTS) were studied in this diploma thesis, using steady-state and time-resolved fluorescence spectroscopy. Two cationic surfactants, Septonex and cetyltrimethylammonium bromide (CTAB), were studied. These surfactants were measured in the complex with hyaluronan (1.75 MDa, 1 MDa and 300 kDa). Steady-state fluorescence was used for determination of critical aggregation concentration of each surfactant and pKa*. Time-resolved fluorescence decays were used to calculate the average lifetimes and the deprotonation constants of naphthol and HPTS. The measurement with hyaluronan were compared with the polystyrenesulfonate (PSS) – surfactant system. The effect of hydration shell of hyaluronan on hyaluronan – surfactant complex formation results from the comparison of above mentioned systems. Large differences were found in the deprotonation characteristic between surfactants and even between individual molecular weights of hyaluronan. The measurement shows that the hydration shell is located near to the dissociated carboxyl groups of hyaluronan chain, where the interaction with the positively charged surfactants occurs. Furthermore, the aggregation number of Septonex was determined by quenching of pyrene using cetylpyridinium chloride (CPC) as a quencher. The aggregation number for 20 mM Septonex solution was determined as a value of 104 molecules. CPC was used for confirmation of the localization of -naphthol in the micelles of CTAB and polymer – CTAB, respectively.
The deprotonation characteristics of fluorescent probes -naphthol and 8-hydroxypyrene-1,3,6-trisulphonic acid (HPTS) were studied in this diploma thesis, using steady-state and time-resolved fluorescence spectroscopy. Two cationic surfactants, Septonex and cetyltrimethylammonium bromide (CTAB), were studied. These surfactants were measured in the complex with hyaluronan (1.75 MDa, 1 MDa and 300 kDa). Steady-state fluorescence was used for determination of critical aggregation concentration of each surfactant and pKa*. Time-resolved fluorescence decays were used to calculate the average lifetimes and the deprotonation constants of naphthol and HPTS. The measurement with hyaluronan were compared with the polystyrenesulfonate (PSS) – surfactant system. The effect of hydration shell of hyaluronan on hyaluronan – surfactant complex formation results from the comparison of above mentioned systems. Large differences were found in the deprotonation characteristic between surfactants and even between individual molecular weights of hyaluronan. The measurement shows that the hydration shell is located near to the dissociated carboxyl groups of hyaluronan chain, where the interaction with the positively charged surfactants occurs. Furthermore, the aggregation number of Septonex was determined by quenching of pyrene using cetylpyridinium chloride (CPC) as a quencher. The aggregation number for 20 mM Septonex solution was determined as a value of 104 molecules. CPC was used for confirmation of the localization of -naphthol in the micelles of CTAB and polymer – CTAB, respectively.
Description
Keywords
Ustálená fluorescence, časově rozlišená fluorescence, ESIPT, ESPT, -naftol, HPTS, cetyltrimethylamonium bromid, Septonex, hyaluronan, polystyrensulfonát, deprotonační konstanty, hydratační obal, agregační číslo., Steady-state fluorescence, time-resolved fluorescence, ESIPT, ESPT, -naphthol, HPTS, cetyltrimethylammonium bromide, Septonex, hyaluronan, polystyrene sulfonate, deprotonation rate constants, hydration shell, aggregation number.
Citation
KOTOUČEK, J. Charakterizace koloidních částic pomocí deprotonace v excitovaném stavu za použití pokročilých fluorescenčních technik [online]. Brno: Vysoké učení technické v Brně. Fakulta chemická. 2016.
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Document version
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cs
Study field
Chemie pro medicínské aplikace
Comittee
prof. Ing. Miloslav Pekař, CSc. (předseda)
prof. RNDr. Ivana Márová, CSc. (místopředseda)
doc. Ing. Pavel Diviš, Ph.D. (člen)
doc. Ing. Stanislav Obruča, Ph.D. (člen)
doc. RNDr. Vladimír Velebný, CSc. (člen)
prof. Ing. Oldřich Zmeškal, CSc. (člen)
Date of acceptance
2016-05-30
Defence
Diviš: Ludox – o jakou sloučeninu se jedná
Márová – význam studia v rámci diplomové práce
Result of defence
práce byla úspěšně obhájena
Document licence
Standardní licenční smlouva - přístup k plnému textu bez omezení