magistrsko delo
Tjaša Damijan (Author), Maša Knez Hrnčič (Mentor), Petra Kotnik (Co-mentor), Taja Žitek (Co-mentor)

Abstract

Industrijska konoplja (Cannabis sativa L.) je zdravilna rastlina s poreklom iz Srednje Azije, ki človeštvo spremlja že več tisoč let. V zadnjem času vzbuja pozornost zaradi številnih aplikacij. Konopljino steblo se uporablja kot vir vlaken za gradbeno, papirno, tekstilno in avtomobilsko industrijo, semena konoplje v živilski industriji za pridobivanje konopljinega olja, listi in cvetovi konoplje pa kot vir bioaktivnih sestavin. Danes je vse večji trend uporabe industrijske konoplje v farmacevtske namene. Sorte konoplje Cannabis Sativa L. vsebujejo nizke ravni Δ9-tetrahidrokanabinola (THC) in se lahko uporabljajo za proizvodnjo terapevtskih izvlečkov, bogatih s kanabidiolom (CBD), ki ima širok razpon farmacevtskih učinkov na človeka z izjemnim medicinskim potencialom pri nevroloških boleznih in protitumorskih aplikacijah. V magistrskem delu smo pridobili izvlečke različnih delov industrijske konoplje in raziskali njihove možne inhibicijske učinke na rakaste celice. S sejalnim postopkom industrijske konoplje smo pridobili sedem frakcij materiala iz različnih delov rastlin, jih ekstrahirali z uporabo ultrazvočne in superkritične ekstrakcije ter nato dekarboksilirali. Za izolacijo olja iz frakcije, ki je vsebovala semena konoplje, smo uporabili destilacijo z vodno paro po Clevengerju. Z UV-spektrofotometrijo smo v dobljenih ekstraktih spremljali deleže bioaktivnih komponent v posameznih frakcijah. Analizirali smo vsebnost celokupnih fenolov, flavonoidov in proantocianidinov. Za pridobljene produkte po ekstrakciji smo izvedli LC-MS/MS analizo. Določeni so bili kanabinoidi CBD, CBDA, CBGA, CBC, CBN, THC in THCA. Nazadnje smo preverili učinek ekstraktov na celice človeškega melanoma (WM-266-4) in na podlagi vidnega preskoka oblike celic uspeli določiti mejno koncentracijo, potrebno za zaviranje nadaljnje delitve rakavih celic (c = 0,001 mg/ml). Rezultati so pokazali, da je ekstrakt, pridobljen iz najbolj fine frakcije sejanja (konopljin prah), veliko bogatejši z biološko aktivnimi komponentami kot ekstrakt frakcije bolj grobega mletja (konopljina stebla). Ekstrakti materiala z višjo vsebnostjo kanabinoidov so pokazali večjo inhibicijo rakastih celic kot ekstrakti materiala z nižjo vsebnostjo kanabinoidov. Predpostavljamo, da je visok delež kanabinoidov v izvlečkih bistven za inhibicijo metabolne aktivnosti rakastih celic. Z magistrskim delom smo potrdili, da ima sejanje konoplje pomembno vlogo pri pridobivanju kakovostnega ekstrakta. Opazna je bistvena razlika, predvsem pri vsebnosti CBD-komponente. S sejanjem lahko natančneje določimo, kateri del materiala bomo ekstrahirali, in posledično zmanjšamo porabo topila ter s tem količino odpadkov pri predelavi.

Keywords

industrijska konoplja;kanabinoidi;WM-266-4 celice;sejanje;magistrske naloge;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher: [T. Damijan]
UDC: 66.066:633.522(043.2)
COBISS: 96817155 Link will open in a new window
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Downloads: 52
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Other data

Secondary language: English
Secondary title: Influence of particle size on polyphenols and cannabinoids in hemp extracts
Secondary abstract: Industrial hemp (Cannabis sativa L.) is a medicinal plant native to Central Asia that has accompanied mankind for thousands of years. Recently, it has gained attention due to its numerous applications. Hemp stalks are used as a source of fiber for the construction, paper, textile, and automotive industries; hemp seeds are used in the food industry to produce hemp oil; and hemp leaves and flowers are used as a source of bioactive ingredients. Today, there is a growing trend to use industrial hemp for pharmaceutical purposes. The hemp varieties Cannabis Sativa L. contain small amounts of Δ9-tetrahydrocannabinol (THC), but can be used to produce therapeutic extracts rich in cannabidiol (CBD). CBD has a wide range of pharmaceutical effects on humans with outstanding medicinal potential in neurological diseases and antitumor applications. In present master thesis, extracts from different parts of industrial hemp have been obtained and their possible inhibitory effects on cancer cells have been investigated in the next stage. Material from seven parts of the plant have been fractioned using the sieving screening method. In addition, the separated materials were extracted using ultrasonication and supercritical extraction followed by decarboxylation. Clevenger's method was used to isolate the oil from the fraction containing hemp seeds. The proportions of bioactive components in each fraction of the extracts obtained have been determined by UV spectrophotometry. The content of total phenols, flavonoids and proanthocyanidins has been analyzed. LC-MS/MS analysis has been performed to determine the contentv and the presence of cannabinoids CBD, CBDA, CBGA, CBC, CBN, THC and THCA. Finally, the effect of the extracts on human melanoma cells (WM -266-4) and based on a visible change in cell shape has been investigated. The minimum concentration required to inhibit further division of cancer cells (c = 0.001 mg / ml) has been determined. The results show that the extract obtained from the finest sieve fraction was much richer in content of biologically active components than the extracts from material fractions with larger particles. Extracts from material with higher cannabinoid content showed more significant inhibition of cancer cells than extracts from material with lower cannabinoid content. It has been concluded that a high content of cannabinoids in extracts is essential for inhibiting the metabolic activity of cancer cells. It has been confirmed that the sieving procedure of hemp plays a crucial role in obtaining a high quality extract. There is a significant difference, especially in the content of the CBD component. By sieving the procedure gains on the selectivity by choosing the most potential part of the material to extract and consequently solvent consumption and thus the amount of waste during processing is decreased.
Secondary keywords: industrial hemp;cannabinoids;CBD;WM-266-4 cells;sieving;
Type (COBISS): Master's thesis/paper
Thesis comment: Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages: 1 spletni vir (1 datoteka PDF (XI, 49 f.))
ID: 13902030