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Ekstrakcija zdravilne učinkovine kordicepin iz glive Cordyceps militaris in njegovo določevanje z uporabo visokoločljivostne tekočinske kromatografije
ID Zotler, Mateja (Author), ID Marolt, Gregor (Mentor) More about this mentor... This link opens in a new window

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Abstract
Cordyceps militaris je zdravilna gliva, zelo cenjena v tradicionalni azijski medicini. Njena glavna bioaktivna spojina je kordicepin. Cilj diplomske naloge je bil optimizacija ekstrakcije kordicepina iz posušene in zmlete glive C. militaris. Za spremljanje učinkovitosti ekstrakcije sem delno validirala HPLC analizno metodo za določevanje vsebnosti kordicepina v ekstraktih. Uporabljena analizna metoda se je izkazala za ponovljivo in linearno v delovnem območju. Optimizacije sem se lotila enofaktorsko. Vsak parameter sem optimizirala v samostojni seriji poskusov. Parameter, ki sem ga želela optimizirati, sem pri vsaki ekstrakciji znotraj serije spremenila, medtem ko so bili ostali parametri konstantni. Optimizirano vrednost parametra sem nato uporabila pri nadaljnjih serijah. Uspešnost ekstrakcije sem ovrednotila glede na razmerje med čim večjim deležem ekstrahiranega kordicepina in čim manjšim deležem nečistoč v suhi masi ekstrakta. V diplomskem delu sem izvedla pet večjih skupin poskusov, v katerih sem preučevala: (i) vpliv deleža etanola v topilu na učinkovitost ekstrakcije, (ii) vpliv različnih razmerij med volumnom topila in maso suhega vzorca na učinkovitost ekstrakcije, (iii) izluževanje kordicepina iz vzorca, ki mu je bil zaporedno odstranjen ekstrakt in dodano sveže topilo, (iv) vpliv pH zmesi vzorca in topila na učinkovitost ekstrakcije, ter (v) perkolacije pri različnih temperaturah, pri čemer sem spremljala časovno odvisnost koncentracije kordicepina v eluentu. Najprej sem izvedla optimizacijo ekstrakcije v šaržni obliki. Za najprimernejše topilo se je izkazal 76,8 % (V/V) etanol, za najprimernejše razmerje med volumnom topila in maso glivnega vzorca pa 20 mL/g. Izkazalo se je, da pH ekstrakcijske zmesi topila in glivnega vzorca ne vpliva na končno učinkovitost ekstrakcije. V delno šaržni obliki sem izvedla tudi imitacijo industrijske ekstrakcije, pri kateri bi v reaktorju macerirali topilo in posušen C. militaris, nato pa bi odlili ekstrakt nad usedlino in dodali sveže topilo. Ob vsakem naslednjem dodatku topila se koncentracija v ekstraktu zmanjša za približno en velikostni razred. Za najučinkovitejšo se je izkazala perkolacija pri 60 °C.

Language:Slovenian
Keywords:kordicepin, ekstrakcija, Cordyceps militaris, optimizacija procesa, HPLC
Work type:Bachelor thesis/paper
Typology:2.11 - Undergraduate Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2023
PID:20.500.12556/RUL-149597 This link opens in a new window
COBISS.SI-ID:169953283 This link opens in a new window
Publication date in RUL:07.09.2023
Views:317
Downloads:42
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Secondary language

Language:English
Title:Extraction of active substance cordycepin from Cordyceps militaris and its determination using high-performance liquid chromatography
Abstract:
Cordyceps militaris is a medicinal fungus highly valued in traditional Asian medicine, with cordycepin as its main bioactive compound. The aim of the thesis was to optimize cordycepin extraction from dried powdered C. militaris. For monitoring the efficiency of the extraction, I partially validated an HPLC analytical method for determining cordycepin in extracts. Analytical method has been proven to be reproducible and linear within the working range. I approached the optimization through single-factor experiments. Each parameter was optimized in a separate series of experiments, where the parameter of interest was varied while keeping the other parameters constant. The optimized value of the parameter was then used in further series of experiments. The extraction efficiency was evaluated based on the ratio of extracted cordycepin and the percentage of impurities in the dry extract. I conducted five major groups of experiments, studying the influence certain parameters have on extraction efficiancy: (i) ethanol concentration in the solvent, (ii) different ratios of solvent volume to dry sample mass, (iii) leaching of cordycepin from a sample from which the extract was sequentially removed and fresh solvent added, (iv) pH of the sample-solvent mixture, and (v) percolation at different temperatures, while monitoring the time-dependent concentration of cordycepin in the eluent. First, I conducted batch-wise extraction optimization. The most suitable solvent was found to be 76.8% (V/V) ethanol, and the most suitable ratio of solvent volume to fungus sample mass was 20 mL/g. It was found that the pH of the extraction mixture of solvent and fungus sample did not affect the final extraction efficiency. I also conducted an imitation of industrial extraction in a semi-batch form, whereas the solvent and dried C. militaris would be macerated in an extraction vessel, and then the extract decanted over the sediment and fresh solvent added. I found that with each subsequent addition of the solvent, the concentration in the extract decreases by approximately one order of magnitude. Percolation at 60 °C was found to be the most efficient.

Keywords:cordycepin, extraction, Cordyceps militaris, optimization of process, HPLC

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