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Ajda (Fagopyrum sp.) je rastlina, ki jo že vrsto let gojimo za uporabo v prehrani in farmaciji. V zadnjem času njeni koristni učinki na zdravje ljudi prihajajo še bolj v ospredje. Poleg visokega deleža proteinov in mineralov je znana po veliki vsebnosti fenolnih spojin, predvsem rutina, ki ga največ najdemo v njeni zeli. Njena raznolika sestava bioaktivnih snovi pozitivno vpliva na potek pogostih bolezenskih stanj, kot so povišan krvni tlak, hiperholesterolemija in sladkorna bolezen. V raziskavah so dokazali tudi nevroprotektivno, protitumorno in protivnetno delovanje. V magistrski nalogi smo ugotavljali vsebnosti flavonoidov rutina, kvercetina in kvercitrina ter fagopirinov v ekstraktih navadne ajde (Fagopyrum esculentum) sorte Darja, pridobljenih z ekstrakcijo s superkritičnim CO2 in etanolom. Vsebnost proučevanih spojin smo ugotavljali v ekstraktih listov, cvetov, nezrelih in zrelih plodov ter ajdovi kaši z analizo konvergenčne kromatografije ultravisoke ločljivosti (UPC2) in tekočinske kromatografije visoke ločjlivosti (HPLC). Za podrobnejši vpogled v sestavo vzorcev smo izvedli tudi analizo z GC-MS. Flavonoidov nismo detektirali pri vseh vzorcih. Največjo vsebnost (52,54 mg/g droge) smo ugotovili pri suhih listih navadne ajde. Od vseh treh flavonoidov se je najpogosteje pojavljal kvercitrin, in sicer največ v etanolnem ekstraktu cvetov frakcije CV1 (2,63 mg/g droge). Izmed CO2 in etanolnih ekstraktov je imel največ rutina etanolni ekstrakt listov frakcije CV1 (0,23 mg/g droge), največ kvercetina pa etanolni ekstrakt listov frakcije CV3 (0,50 mg/g droge). Povzamemo lahko, da so pri navadni ajdi (F. esculentum) sorte Darja s flavonoidi najbolj bogati listi in cvetovi, najrevnejša pa kaša. V nasprotju s flavonoidi smo fagopirine detektirali pri vseh vzorcih. Največjo vsebnost smo ugotovili v CO2 ekstraktu listov frakcije CV1 (25,54 mg/g droge). Največ spojin (21) smo identificirali pri etanolnem ekstraktu listov frakcije CV3, najmanj pa pri kaši. Največ neidentificiranih snovi (18) smo analizirali pri etanolnem ekstraktu cvetov frakcije CV1. Delež identificiranih snovi je bil skoraj pri vseh analiziranih ekstraktih, razen pri kaši, okoli 80 %. Buckwheat (Fagopyrum sp.) is a plant that has been grown for its use in food and pharmacy for years. Lately, it has made it to the forefront with its beneficial effects on human health. Additionally to its high levels of proteins and minerals it is known for its high content of phenolic compounds, especially rutin, which is mostly found in its herb. Its diverse composition of bioactive substances has a positive effect on the development of common disease states like high blood pressure, hypercholesterolemia and diabetes. It is also attributed with neuroprotective, antitumor and antiinflammatory effects. In this master's thesis we determined the flavonoid contents of rutin, quercetin and quercitrin along with fagopyrins found in the common buckwheat (Fagopyrum esculentum) extracts of the Darja variety, that were obtained by extraction with supercritical CO2 and ethanol. The content of the studied compounds was determined in the extracts of leaves, flowers, unripened and ripened fruits as well as in buckwheat pulp with UPC2 and HPLC analyses. We also performed a GC-MS analysis for a more detailed insight into the composition of the samples. The flavonoids were not detected in all samples. The highest content (52,54 mg/g of the drug) was determined in dry leaves of common buckwheat. Out of all three flavonoids, we most frequently measured the content of quercitrin namely in the ethanolic flower extract of the CV1 fraction (2,63 mg/g of the drug). Out of the CO2 and ethanol extracts, the most rutin is in the ethanolic leaves extract of the CV1 fraction (0,23 mg/g of the drug), and the most of the quercetin is in the ethanolic leaves extract of the fraction CV3 (0,50 mg/g of the drug). We can summarize that in the common buckwheat (F. esculentum) of the Darja variety, leaves and flowers come in with the highest flavonoid levels while the pulp comes in last with the lowest amounts. In contrast to flavonoids, fagopyrins were detected in all samples. The highest content was found in the CO2 leaves extract of the CV1 fraction (25,54 mg/g of the drug). Most compounds (21) were identified in the ethanolic leaves extract of the CV3 fraction, and at the least here in the pulp. Most unidentified substances (18) were analyzed in the ethanolic flower extract of the CV1 fraction. The proportion of identified substances in almost all analyzed extracts, except for pulp, is about 80 %.

V magistrskem delu smo določali protiglivno aktivnost izvlečkov rastlinskih vrst Chamomilla recutita, Helichrysum arenarium, Humulus lupulus, Juniperus communis, Achillea millefolium, Calendula officinalis, Sambucus nigra, Nepeta cataria, Olea europaea, Hypericum perforatum, Taraxacum officinale, Crataegus sp., pripravljenih z ekstrakcijo s superkritičnim ogljikovim dioksidom. Njihovo protiglivno delovanje smo določali proti vrstam Epicoccum nigrum, Alternaria alternata, Fusarium poae, Fusarium oxysporum in Botrytis cinerea tako, da smo iz koncentriranih rastlinskih izvlečkov pripravili 10 % raztopine, ki smo jih dodali na 2 % PDA-gojišče in nanje neposredno nacepili kulturo glive. Vse tretmaje smo trikrat ponovili. Vsi rastlinski izvlečki, pripravljeni z ekstrakcijo s superkritičnim ogljikovim dioksidom, so zavrli rast gliv. Največji zaviralni učinek na rast gliv sta imela izvlečka C. recutita in H. arenarium. Na izvlečke je bila najbolj občutljiva gliva B. cinerea 5A, največja odpornost proti izvlečkom pa je bila ugotovljena pri B. cinerea 4A. Za dokazovanje protiglivnega delovanja izvlečkov kamilice in smilja smo glive gojili na izvlečkih s koncentracijami 50 %, 25 %, 12,5 %, 6,25 % in 3,125 % ter pri tem ugotovili, da je bila inhibicija rasti koncentracijsko odvisna. Rezultati raziskave so pokazali, da imajo izvlečki, pridobljeni z ekstrakcijo s superkritičnim ogljikovim dioksidom, potencial za uporabo v živilski industriji in medicini pri zaviranju rasti škodljivih gliv. In this master's thesis we tested the biological activity of extracts from plant species Chamomilla recutita, Helichrysum arenarium, Humulus lupulus, Juniperus communis, Achillea millefolium, Calendula officinalis, Sambucus nigra, Nepeta cataria, Olea europaea, Hypericum perforatum, Taraxacum officinale and Crataegus sp. prepared by extraction with supercritical carbon dioxide. We tested for antifungal activity against species Epicoccum nigrum, Alternaria alternata, Fusarium poae, Fusarium oxysporum and Botrytis cinerea, by creating a 10% concentration of the plant extracts, which were then pipetted on the center of the petri dish with a 2% PDA gojišče to which we added the fungi culture. All treatments were performed in three replicates. All extracts prepared with supercritical carbon diokside inhibited fungal growth. The biggest impact on growth was observed in extracts of C. recutita and H. arenarium. The highest sensitivity to treatments was observed in the fungi B. cinerea 5A, while the highest resistance to extracts was observed in B. cinerea 4A. To demonstrate antifungal activity in the most effective plant extracts, C. recutita and H. arenarium, fungi were grown on extracts in concentrations of 50 %, 25 %, 12.50 %, 6.25 % and 3.125 %. We found out that the inhibition of fungal species is concetration dependant. The results of study showed, that extracts obtained with supercritical carbon dioxide have great potential for use in the food industry and medicine in inhibiting the growth of harmful fungi.

V diplomskem delu sem prikazal teoretične osnove superkritičnih tekočin in superkritične ekstrakcije. Kritična točka ima kritično temperaturo in kritičen tlak. Superkritične tekočine so v superkritičnem območju, to pomeni, da sta temperatura in tlak višja od tistih v kritični točki. Pri ekstrakciji odstranjujemo topne komponente iz trdnih ali tekočih zmesi. Proces temelji na razliki v topnosti komponent v zmesi, pridobljena snov pa se imenuje ekstrakt. Pri superkritični ekstrakciji se namesto navadnih organskih topil uporabljajo superkritične tekočine. S tem dosežemo hitrejšo ekstrakcijo in boljšo selektivnost. Ekstrahiramo lahko tekočine, trdne snovi, maščobe in olja, rastlinske materiale, vode in soli. V svojem diplomskem delu sem opisal superkritične tekočine, parametre, ki so povezani z učinkovitostjo procesa, sam proces in uporabo v industriji. In my thesis, I presented theoretical basics of supercritical fluids and supercritical fluid extraction. Critical point possesses critical temperature and critical pressure. Supercritical fluids exist in supercritical zone, where temperature and pressure are higher than ones in critical point. Liquids, solids, fats and oils, plant materials, water and salts can be extracted. During extraction process, soluble components are removed from solid or liquid mixtures. The extraction is based on the difference in the solubility of the components in the mixture, and the substance we get in the end is called the extract. Supercritical fluids are used as solvent instead of ordinary organic solvents in supercritical extraction. This benefits in better selectivity and faster extraction. I have described supercritical fluids, the parameters associated with process efficiency, the process itself, and its use in industry.

U okviru ove disertacije sprovedena suispitivanja valorizacije biljne prašine herbehajdučke trave i ploda šipka, kao sporednihproizvoda nastalih u toku proizvodnje biljnogfilter čaja, u smislu implementacije u noveproizvode, s obzirom na to da se radi o značajnojkoličini biljnog sporednog proizvoda/otpada i dase pri tome radi o vrednoj sirovini koja i daljesadrži određenu količinu značajnih bioaktivnihjedinjenja. Najpre je izvršena karakterizacijabiljnog materijala, a zatim, primenom različitihtehnika, i njihova transformacija u biljnepreparate-ekstrakte. U cilju pripreme biljnihpreparata-ekstrakata, primenjene su triekstrakcione tehnike: klasična ekstrakcijamaceracija,sokslet ekstrakcija i ekstrakcijasuperkritičnim ugljen-dioksidom, a pored togaza transformaciju iz tečnog u suvu formuprimenjena je tehnika sušenja sa raspršivanjemspraydrying. Primenom različitih metodaanalize utvrđen je sastav dobijenih ekstrakata.Ispitana je, takođe, i mogućnost inkorporacijesporednog proizvoda kao i dobijenih biljnihpreparata u nove proizvode, te je u tom smislu,pripremljen i ispitan instant čaj od suvogekstrakta hajdučke trave, dok se biljna prašinaploda šipka uspešno iskoristila za dobijanjefunkcionalnog hleba., As part of this dissertation, the valorisationstudies of herb dust of Achillea millefolium andRosa canina L fruit, as by-products of plantfilter tea production, were conducted in terms ofimplementation into new products, given that itis a large amount of plant by-product / waste,and that it is a valuable raw material that stillcontains a certain amount of significantbioactive compounds. First, the characterizationof the plant material was performed, and then,using various techniques, their transformationinto herbal preparations-extracts. In order toprepare herbal preparations-extracts, threeextraction techniques were applied: classicalextraction-maceration, soxhlet extraction, andsupercritical carbon dioxide extraction, and inaddition, the spray-drying technique was appliedfor the transformation from liquid to dry form.The composition of the obtained extracts wasdetermined using various methods of analysis.The possibility of incorporating the by-productas well as the obtained herbal preparations intonew products was also examined, thereforeinstant tea was prepared and tested from the dryextract of A.millefolium, while the herb dust ofthe R.canina L fruit was successfully used toobtain functional bread.

Namen diplomske naloge je bil raziskati potencialne protimikrobne in antioksidativne učinke različnih ekstraktov, pridobljenih iz ingverja (Zingiber officinale), s ciljem potencialne uporabe naravnih izvlečkov v prehrambeni, farmacevtski, živilski in kozmetični industriji. Ekstrakte smo pridobili z različnimi metodami ekstrakcije s Soxhletovo metodo, ultrazvočno metodo in superkritično ekstrakcijo s CO2. Vse ekstrakcije smo izvajali dve uri. Pri Soxhletovi in ultrazvočni ekstrakciji smo uporabili različna topila, s ciljem prepoznati razliko med ekstrahiranimi komponentami s pomočjo polarnega in nepolarnega topila. Različne metode ekstrakcije in različna topila vplivajo na količino pridobljenega ekstrakta in na vsebnost komponent, ki jih ekstrakt vsebuje. Zanimal nas je izkoristek ekstrakcij in kvaliteta pridobljenih ekstraktov. Uporabili smo dve spektrofotometrični metodi radikalsko metodo DPPH za določanje antioksidativne aktivnosti ekstraktov in metodo za določanje vsebnosti totalnih fenolov. Za določevanje protimikrobnega učinka smo uporabili difuzijsko metodo z diski, protimikrobno delovanje smo testirali na grampozitivni bakteriji Staphylococcus aureus, gramnegativni bakteriji Escherichia coli in kvasovki Candida albicans. V diplomskem delu so prikazani rezultati ekstrakcij in analiznih metod, ki smo jih izvedli. Iz rezultatov je vidno, da je uporaba Soxhletove ekstrakcije in topila aceton najboljša izbira za dosego najvišjega izkoristka ekstrakcije. Antioksidativni potencial smo ovrednotili z DPPH metodo, s katero smo najvišji odstotek inhibicije opazili pri ekstraktu, pridobljenem s superkritično ekstrakcijo pri 135 bar. Najnižji odstotek inhibicije smo opazili pri obeh ekstraktih, pridobljenih s topilom heksan. Pri določevanju vsebnosti totalnih fenolov smo opazili najvišjo vsebnost v ekstraktu, pridobljenem z ultrazvočno ekstrakcijo, kjer smo kot topilo uporabili mešanico acetona in vode v razmerju 1:1. Najnižji delež vsebnosti totalnih fenolov smo opazili pri ekstraktoma, pridobljenima s topilom heksan kar sovpada z najnižjim antioksidativnim potencialom. Nekateri pridobljeni ekstrakti so delovali protimikrobno proti C. albicans in S. aureus, medtem ko proti E.coli noben ekstrakt ni pokazal protimikrobnega učinka. The purpose of the thesis was to research potential antimicrobial and antioxidant effects of different extracts from ginger (Zingiber officinale) with the aim of potential use of natural extracts in nutritional, pharmaceutical, food and cosmetic industry. The extracts were obtained by different extraction methods Soxhlet extraction, ultrasound extraction and supercritical CO2 extraction. Each extraction lasted two hours. We used different solvents by Soxhlet extraction and ultrasound extraction with the aim to get know the difference among extracted components by using polar and nonpolar solvent. Different extraction methods and different solvents have impact on the quantity of obtained extract and the content of extract components. We were interested in extractions efficiency and the quantity of obtained extracts. We used only two spectrophotometric methods DPPH radical method to determine antioxidant activity of extracts and the method to determine total phenolic content. We used disk diffusion method for determining antimicrobial efficiency, antimicrobial activity was tested on gram positive bacteria Staphylococcus aureus, gram negative bacteria Escherichia coli and Candida albicans yeast. In the thesis we show the results of extractions and analytical method that we derived. It is proved by the results that Soxhlet extraction and acetone as a solvent are the best choice to obtain the highest extraction efficiency. Antioxidant potential was determined by DPPH method which revealed the highest percentage of inhibition extract that has been obtained by supercritical extraction at 135 bar. The lowest percentage of inhibition was noticed in both extracts obtained by hexane as a solvent. By determining total phenolic content we noticed the highest content in the extract which was obtained by ultrasound extraction, there we used acetone-water mixture in the ratio 1:1. The lowest percentage of total phenolic content was in extracts obtained by hexane as a solvent, the reason lies in the lowest antioxidant potential. Some obtained extracts were antimicrobial against C. albicans and S. aureus, on the other hand no extract did not have antimicrobial effect on E.coli.

Povzetek V diplomskem delu smo raziskali, če izbrana živila vsebujejo vitamin D2 (ergokalciferol), provitamin D2 (ergosterol), vitamin D3 (holekalciferol) in provitamin D3 (7-dehidroholesterol). Analizirali smo živila rastlinskega izvora: ričkovo olje, konopljino olje, posušeno konopljo, gobe (bukov ostrigar) in lucerno ter živila živalskega izvora: sir, sirotka in jajce (rumenjak in beljak). Živila smo predhodno pripravili za nadaljnje analize. Konopljo in suho lucerno smo zmleli, svežo lucerno in sveže gobe smo liofilizirali, nato zmleli, preostanek svežih gob narezali in zamrznili, prav tako tudi svežo lucerno. Sir in sirotka sta bila proizvedena na Kmetiji Krevh po postopku izdelave poltrdega sira. Jajca proste reje smo uporabili sveža. Konopljo, lucerno in gobe smo ekstrahirali po metodi ekstrakcije po Soxhletu in ekstrakcije s superkritičnim fluidom (CO2). Materialom smo določili vsebnost prostih maščobnih kislin. Ekstrakte in ostale materiale smo saponificirali po postopku za saponifikacijo zaestrenega vitamina D in nato vse vzorce (saponificirane in nesaponificirane) analizirali s tekočinsko kromatografijo z masno spektrometrijo. Znano je, da je vitamin D2 prisoten v rastlinskih živilih, vitamin D3 pa v živalskih. Iz nekaterih virov smo ugotovili, da lahko tudi rastline vsebujejo vitamin D3, ter živalski proizvodi vitamin D2, kar smo potrdili tudi z našimi analizami. Ugotovili smo, da je za določitev vitaminov in provitaminov D potrebna saponifikacija, saj je vsebnost vitamina v saponificiranih vzorcih višja. Rezultati kažejo, da je količina vitamina D, ki je topen v maščobah, povezana z vsebnostjo maščob v materialu, za nekatere primere pa smo sklepali, da je vitamin vezan na proteinske molekule. The diploma thesis examines if the selected food products contain vitamin D2 (ergocalciferol), provitamin D2 (ergosterol), vitamin D3 (cholecalciferol), and provitamin D3 (7-dehydrocholesterol). We have analysed plant-based foods: Camelina sativa oil, hemp oil, dried hemp, oyster mushrooms, and alfalfa, and animal-based foods: cheese, whey, and eggs (the yoke and the white). We prepared the foods for further analyses. The hemp and dried alfalfa were ground, the fresh alfalfa and fresh mushrooms were freeze-dried and ground, the remaining fresh mushrooms were sliced and frozen, as was the fresh alfalfa. The cheese and whey were produced by the Krevh farm according to the process of producing semi-hard cheese. Free-range eggs were used fresh. The hemp, alfalfa, and mushrooms were extracted using Soxhlet extraction and supercritical fluid extraction (CO2). We determined the content of free fatty acids in the materials. The extracts and other materials were saponificated using the process of saponificating esterificated vitamin D. All samples (both saponificated and non-saponificated) were analysed using fluid chromatography with mass spectrometry. It is known that vitamin D2 is present in plant-based foods, while vitamin D3 is present in animal-based foods. In some sources we found that even plants may contain vitamin D3 and that animal products may contain vitamin D2, which our analyses also confirmed. We found that saponification is required to establish the presence of vitamins and provitamins D, because the vitamin content in saponificated samples is higher. Results indicate that the amount of fat-soluble vitamin D is correlated to the fat content of the material, while for some examples we assumed that the vitamin is bound to protein molecules.

Industrijska konoplja (Cannabis Sativa L.) znova pridobiva svoj pomen v svetu kmetijstva in zdravstva. V zadnjih nekaj letih je pridelovanje industrijske konoplje in izdelkov iz nje zelo naraslo. Danes lahko najdemo veliko kmetij, ki industrijsko konopljo redno pridelujejo in prodajajo njeno seme, listje, cvetje in tudi izdelke, pridelane iz nje, kot so eterična olja in smole. V diplomskem delu smo raziskali različne načine izločanja kanabinoidov in ostalih biološko aktivnih komponent iz posušenih in zmletih lističev industrijske konoplje. Poskusili smo ekstrahirati kanabinoide, kot so CBD, CBN, CBC in njihove kisline CBDA ter CBGA. Našteti kanabinoidi se velikokrat pojavljajo tako v zdravstvu, kot tudi pri osebni uporabi. Raziskave kažejo, da naj bi predvsem CBD in CBG imela pozitiven učinek pri zmanjševanju bolečin in zdravljenju raka. Kanabinoide smo ekstrahirali s konvencionalnimi metodami z uporabo različnih topil in z nekonvencionalnimi metodami z uporabo superkritičnega ogljikovega dioksida (CO2). Izvedli smo ekstrakcije s Soxhletovo aparaturo, ultrazvočne ekstrakcije in maceracije z uporabo različnih topil metanola, etanola in heksana. Ekstrakcije s superkritičnim CO2 smo izvedli v avtoklavu pri različnih temperaturnih in tlačnih pogojih, pri nekaterih smo uporabili tudi kotopilo metanol. Ekstrakte smo nato analizirali z LC-MS metodo, kjer smo določili vsebnost posamezne kanabinoidne komponente. S Clevengerjevo metodo smo izolirali eterično olje iz industrijske konoplje ter s pomočjo GC-MS določili prisotnost terpenskih komponent. S Clevengerjevo metodo smo poskusili pridobiti nekaj eteričnega olja, izkoristek je bil majhen, saj smo pridelali 0,4 ml olja iz 30 g konoplje. Industrial hemp (Cannabis Sativa L.) is gaining its importance in the world of agriculture and health. In the last few years, the production of industrial cannabis and its products has increased considerably. Today, many farms can be found that produce industrial cannabis regularly and sell its seeds, leaves, flowers and also products made from it, such as essential oils and resins. In this work we studied various methods of extraction of cannabinoids and other biologically active components from dried and grounded cans of industrial cannabis. We aimed to extract cannabinoids, such as CBD, CBN, CBC, and their acids CBDA and CBGA. These cannabinoids are present in healthcare as well as in personal use. Research suggests that CBD and CBG should have a positive effect in reducing pain and treating cancer. Canabinoids were extracted with conventional methods using various solvents and non-conventional methods using supercritical carbon dioxide (CO2). Extractions were performed in various ways with Soxhlet's apparatus, ultrasonic extraction and macerating extractions using methanol, ethanol and hexane as a solvent. Supercritical CO2 extractions were carried out in autoclave at different temperature and pressure conditions, some with use of ethanol as cosolvent. The extracts were then diluted and analyzed using the LC-MS method for the content of cannabinoids. The presence of target compounds in extracts, attaned by different extraction methods and solvents has been considered . With Clevenger's method essential oil were extracted, whereas the yield was relatively low, as we collected 0.4 ml oil from 30 g of hemp.

Za izolacijo biološko aktivnih komponent uporabljamo postopek ekstrakcije. Namen je bil ugotoviti kako izbira topila in procesni pogoji vplivajo na kvaliteto oziroma na sestavo ekstrakta pridobljenega iz navadne ajde (Fagopyrum esculentum). Pri eksperimentalnem delu smo uporabili konvencionalne in superkritične metode ekstrakcije. Izvedli pa smo tudi ekstrakcijo polisaharidov. Pridobljenim ekstraktom smo določevali vsebnost prostih ogljikovih hidratov s fenol-žveplovo metodo in vsebnost proteinov z Bradfordovo metodo ter antioksidativno aktivnost ekstraktov z uporabo 2,2-difenil-1-pikrilhidrazil (DPPH*) prostega radikala. Izvedli smo tudi analize LC-MS/MS in določili vsebnost taninov. Najvišji izkoristek smo pridobili z ekstrakcijo po Soxhletu z uporabo topil petroleter in heksan. Ugotovili smo, da v našem primeru masni izkoristek ekstrakcije ne narašča v smeri rasti polarnosti topil. Razlog je v visoki vsebnosti nepolarnih komponent v semenih navadne ajde. Na izkoristek je vplivala tudi višja temperatura pri ekstrakciji. Najvišjo vsebnost prostih ogljikovih hidratov smo zasledili pri ekstraktu, kjer smo ekstrahirali polisaharide in pri nepredelanem materialu. Ugotovili smo tudi, da največ proteinov vsebuje ekstrakt, pridobljen s superkritično ekstrakcijo, najmanj pa sam material. Najvišji % inhibiranega DPPH sta vsebovala ekstrakta, kjer smo kot topilo uporabili metanol in metodo po Soxhletu ter ultrazvočno ekstrakcijo. Ugotovili smo, da temperatura ne vpliva bistveno na antioksidativno aktivnost. Izvedli smo analizo taninov v naših vzorcih z LC-MS/MS. Hidrolizirajoči tanini so bili prisotni samo v metanolnih in etanolnih ekstraktih. Od teh smo identificirali le elagno in galno kislino, ki smo ju tudi kvantificirali. Zaključimo lahko, da je velik delež spojin v ajdi nepolarne narave, ter da njihova topnost narašča z naraščujočo temperaturo. Ajda je kvalitetno živilo s številnimi pozitivnimi učinki na zdravje, saj vsebuje veliko biološko aktivnih snovi, ki smo jih tudi dokazali. The extraction process is used for the isolation of biologically active components. The aim was to determine how the solvents and process conditions influenced the quality and composition of the extracts obtained from the common buckwheat (Fagopyrum esculentum). In the experimental work, conventional and supercritical extraction methods were used. We also carried out the extraction of polysaccharides. In the obtained extracts we determined the free carbohydrate content by the phenolic-sulfur method and the protein content by the Bradford method. Antioxidant activity was determined using the free radical 2,2-Diphenyl-1-picrylhydrazyl (DPPH*). We also carried out an analysis of LC-MS/MS in terms of tannin content. The highest yield was obtained by extraction with Soxhlet using solvent petroleum ether and hexane. In our case the mass yield of the extraction does not increase in the direction of the growth of solvents polarity. The reason is the high content of nonpolar components in common buckwheat seeds. The efficiency was also influenced by the higher temperatures. The highest content of free carbohydrates was found in the extract, where polysaccharides were extracted and in unprocessed material. The highest content of proteins was found in an extract obtained by supercritical extraction, and the lowest content of proteins was in the material itself. The highest % of the inhibited DPPH contained extracts, where methanol as solvent and Soxhlet method and ultrasonic extraction were used. We found that the temperature does not significantly affect the antioxidant activity. We analyzed the tannins in our samples with LC-MS/MS. Hydrolyzable tannins were present only in methanolic and ethanolic extracts. Of these, only ellagic and gallic acid were identified and quantified. It can be concluded that a large proportion of the compounds in the common buckwheat are of nonpolar nature and that their solubility increases with increasing temperature. Common buckwheat has many positive effects on health, as it contains many biologically active substances, which we have also proved.

Glavni cilj ove disertacije koji obuhvata dobijanje i hemijsku karakterizaciju ekstrakata i etarskih ulja rtanjskog čaja (Satureja montana L.), bosiljka (Ocimum basilicum L.) i korijandera (Coriandrum sativum L.), kao i njihove smeše, je određivanje biološke aktivnosti dobijenih ekstrakata u in vitro uslovima pradenjem sposobnosti neutralizacije slobodnih radikala i antiproliferativne aktivnosti. Prvo je izvršena karakterizacija biljnih sirovina (rtanjski čaj, bosiljak i korijander), kao i njihove smeše jednakih masenih udela (CBS) u pogledu sadržaja vlage, srednjeg prečnika čestica i sadržaja etarskog ulja. U cilju dobijanja etarskih ulja i ekstrakata iz ispitivanih sirovina su primenjeni hidrodestilacija (konvencionalni postupak) i superkritična ekstrakcija (savremena tehnika). GC-MS analizom je određen kvalitativni sastav i relativni udeo pojedinačnih monoterpenskih ugljovodonika, oksidovanih monoterpena, seskviterpena i ostalih jedinjenja. Dalje je GC-FID analizom kvantitativno određen sadržaj i prinos pojedinačnih terpena (α-pinen, β-pinen, d-limonen, γ-terpinen, eukaliptol, linalool, kamfor, α-terpineol, geraniol, metil-havikol, karvakrol i eugenol) i izvršeno je poređenje uzoraka dobijenih tradicionalnim i savremenim postupcima. Etarskim uljima i lipofilnim ekstraktima je određena biološka aktivnost ispitivanjem antioksidativne i antiproliferativne aktivnosti. In vitro anti-oksidativna aktivnost superkritičnih ekstrakata i etarskih ulja je određena DPPH testom, dok je antiproliferativna aktivnost određena u in vitro uslovima na tri linije tumorskih delija (HeLa, MDA-MB-453, K562) i na normalne humane fibroblaste (MRC-5). U cilju boljeg određivanja sličnosti između etarskih ulja i lipofilnih ekstrakata rtanjskog čaja, bosiljka, korijandera i njihove smeše (CBS) su primenjene hemometrijska tehnike: analiza glavnih komponenata (Principal component analysis) i hijerarhijska klaster analiza (Hierarchical cluster analysis). The main aim of this dissertation was recovery and chemical characterization of extracts and essential oils of winter savory (Satureja montana L.), basil (Ocimum basilicum L.) and coriander (Coriandrum sativum L.), as well as their mixture, and determination biological activity of extracts in in vitro conditions by capacity towards neutralization of free radicals and antiproliferative activity. Plant materials (winter savory, basil and coriander) and their mixture (CBS) were characterized in terms of moisture content, mean particle size and essential oil content. In order to obtain essential oil and extracts from investigated plants, hydrodistillation (conventional procedure) and supercritical fluid extraction (novel technique) were applied. GC-MS analysis was used for determination of qualitative content and relative abundance of monoterpene hydrocarbons, oxygenated monoterpenes, sesquiterpenes and other volatile compounds. Furthermore, GC-FID analysis was used for quantitative determination of content and yield of terpene compounds (α-pinene, β-pinene, d-limonene, γ-terpinene, eucalyptol, linalool, camphor, α-terpineol, geraniol, methyl-chavicol, carvacrol i eugenol) and samples obtained by traditional and novel techniques were compared. Biological potential of essential oil and lipophilic extracts was determined by antioxidant and antiproliferative activity. In vitro antioxidant activity of supercritical extracts and essential oils was determined by DPPH assay, while antiproliferative activity was determined in in vitro conditions on three cancer cell lines (HeLa, MDA-MB-453, K562) and on normal human fibroblasts (MRC-5). Chemometric techniques such as principal component analysis (PCA) i hierarchical cluster analysis (HCA) were applied for better understanding of similarities between essential oils and lipid extracts.

V okviru doktorske disertacije smo predstavili koristi tehnologij s superkritičnimi fluidi v različnih industrijskih procesih. Čeprav je superkritični CO2 trenutno eden najpogosteje zastopanih superkritičnih fluidov v laboratorijskem merilu, pa je njegova prisotnost na področju industrije še vedno omejena, saj takšna vrsta tehnologije zahteva potrebe po zelo visokih investicijskih stroških. Kljub temu so procesi, kjer se uporablja superkritični CO2 preprosti, okolju prijazni, cena CO2 pa relativno nizka. Takšni procesi predstavljajo odlično alternativo organskim topilom. Postopki ekstrakcije in ločevanja zelo čistih aktivnih substanc, kot sta ksantohumol in beta-sitosterol za zahteve bodisi farmacevtske bodisi prehrambne industrije, kažejo odlično priložnost uporabe procesov s superkritičnim CO2 brez uporabe organskih topil. Doktorska disertacija je razdeljena na tri različne dele. V prvem delu smo raziskovali konvencionalne ekstrakcije in ekstrakcije s superkritičnimi fluidi iz jagod žago-listne palme. V nadaljevanju smo predstavili ter med seboj primerjali konvencionalne kromatografske tehnike s superkritično kromatografijo. Uspešno smo s pomočjo superkritične kromatografije ločili ksantohumol iz hmeljskega ekstrakta. V zadnjem delu doktorske disertacije smo predstavili različne tehnike emulgiranja in enkapsulacije. Ti procesi so zelo pomembni v prehrambni in farmacevtski indistriji, saj je večina aktivnih komponent netopna v vodi. Takšni procesi omogočajo proizvodnjo takšnih formulacij, ki se nato topijo v ustreznih medijih. Eksperimentalni del doktorske disertacije obsega ekstrakcije iz jagod žago-listne palme pri višjih obratopvnih pogojih kot so bili uporabljeni v predhodnih študijah. Ločevanje ksantohumola smo izvedli na pilotni napravi za superkritično kromatografijo, kjer smo spreminjali temperaturo, tlak, masni pretok, mobilno in stacionarno fazo. Rezultate smo nato primerjali s tistimi pridobljenimi s konvencionalnimi tehnikami. Skušali smo dokazati prednosti uporabe superkritične kromatografije pred drugimi tehnikami. This doctoral dissertation is focused on the benefits of supercritical fluids technology in several industry processes. Although, supercritical CO2 is currently the most employed supercritical fluid at laboratory scale, its presence in the industry field is still limited, due to the necessity of high investment costs. Despite that fact, operations with supercritical CO2 are cheap, environmentally friendly and simple, which make it an excellent alternative to organic compounds. Extraction and separation of highly pure active compounds, such as xanthohumol or beta-sitosterol, are demanded for pharmaceutical or food industries, which create an excellent opportunity for supercritical CO2 operations, since they are solvent free. This manuscript is divided into three different parts. In the first part, conventional extraction processes and supercritical fluid process are theoretically compared and supercritical fluid extraction from saw palmetto berries is presented. In the second part, conventional chromatographic techniques and supercritical fluid chromatography are theoretically compared and separation of xanthohumol from hop extracts by supercritical fluid chromatography is presented. In the last part, different encapsulation and emulsification processes are listed and defined. These processes are very important for food and pharmaceutical companies, since several active compounds are not water soluble which create the necessity of different formulations in order to be solved in an appropriate media. Regarding the experimental part, extractions from saw palmetto berries were performed at various pressures and temperatures, which were higher than the ones that were applied in previous studies, in order to estimate and calculate different parameters (solubility, extraction kinetic curves, composition,…), and to compare different studies. Separation of xanthohumol was carried out in a homemade supercritical chromatography, which manageable variables were temperature, pressure, mass flow-rate, modifier, and stationary phase. The results were compared with those ones which were obtained using conventional techniques, in order to prove the suitability and the improvement of this technique against others tecniques.

U okviru ove disertacije su ispitane mogućnosti primene savremenih postupaka za dobijanje ekstrakata rtanjskog čaja (Satureja montana L.).U ekstraktima dobijenim primenom superkritičnog ugljendioksida (različitih gustina) utvrđeno je prisustvo karvakrola kao najzastupljenije komponente. Ekstrakt sa najvećim sadržajem karvakrola je dobijen pri uslovima 350 bar i 50 °C (60,82%), dok je najveći prinos karvakrola ostvaren pri pritiscima 325 i 350 bar i temperaturi 60 °C, i iznosio je 2,4 g/100 g droge. Primenom ugljendioksida na većim pritiscima se ostvaruje efikasnija ekstrakcija karvakrola iz droge, kao i proizvodnja ekstrakata sa većim sadržajima karvakrola. Druge komponente prisutne u ekstraktima u znatno nižim koncentracijama su: p-cimen, borneol, trans-kariofilen, kariofilen-oksid, γ-terpinen i linalool.Da bi se utvrdila mogućnost unapređenja prinosa ekstrakcije superkritičnim ugljendioksidom, kao i poboljšanja kvaliteta ekstrakata, ispitan je uticaj različitih predtretmana biljnog materijala. Ustanovljeno je da predtretman vodom najznačajnije povećava prinos ekstrakcije (za 25%), dok je ultrazvučni predtretman najadekvatniji izbor za dobijanje ekstrakata sa najvećim sadržajem karvakrola (66,46%). Međutim, najveći prinos ekstrakcije karvakrola iz 100 g droge postignut je primenom etanola i vode kao predtretmana.Ekstrakti dobijeni primenom subkritične vode (uz variranje temperature i vremena ekstrakcije) su hemijski okarakterisani i radi utvrđivanja uslova ekstrakcije pri kojima se postiže najveći kvalitet ekstrakata u pogledu sadržaja polifenolnih komponenti, kao i antioksidantne aktivnosti, primenjena je metoda odzivne površine (RSM). Utvrđena je prednost primene subkritične vode u odnosu na klasičnu metodu ekstrakcije u pogledu sadržaja polifenolnih komponenti i antioksidantne aktivnosti.Suvi ekstrakti S. montana dobijeni spray drying tehnologijom sušenja (sa različitim koncentracijama maltodekstrina) su ispitani u pogledu fizičkih i hemijskih osobina. Takođe, radi ispitivanja mogućnosti primene u vidu funkcionalnih ili prehrambenih proizvoda, izvršena je senzorna analiza dobijenih suvih ekstrakata, i analizirana je njihova farmakološka aktivnost u smislu ACE inhibitornog i antioksidantnog delovanja.Ustanovljeno je da je najniža koncentracija maltodekstrina (10%) najadekvatnija za dobijanje suvog ekstrakta S. montana sa najvećim sadržajem polifenolnih komponenti, sadržajem etarskog ulja i u pogledu antioksidantne i ACE inhibitorne aktivnosti. Takođe, suvi ekstrakt sa 10% maltodekstrina ispunjava i ostale fizičko-hemijske parametre, kojima se osigurava kako efikasno sušenje, tako i kvalitet ekstrakta.In vivo aktivnost odabranih ekstrakata S. montana ispitana je na modelu hepatotoksičnosti indukovane primenom ugljentetrahlorida. Analiziran je uticaj ekstrakata na funkciju jetre, enzimske parametre oksidativnog stresa i serumske parametre oksidativnog oštećenja izazvanog ugljentetrahloridom i ustanovljen je hepatoprotektivni učinak ekstrakata S. montana. Takođe, ispitano je dejstvo ekstrakata na rast Ehrlich-ovih ascitnih tumora implantiranih miševima, kao i na biohemijske parametre oksidativnog stresa u ćelijama tumora i utvrđeno je da je vreme aplikacije ekstrakata od izuzetnog značaja., This dissertation investigates the possibilities of applying contemporary processes for obtaining extracts of winter savory (Satureja montana L.).Carvacrol was determined to be the most abundant component in extracts acquired by using supercritical carbon dioxide (of various densities). Extract with the richest content of carvacrol was obtained under conditions of 350 bar and 50 °C (60,82%), while the highest carvacrol yield was produced under 325 bar and 350 bar pressures and at a 60 °C temperature reaching 2.4 g/100 g of dry weight. By applying carbon dioxide on higher pressures, a more efficient carvacrol extraction from herbal material is performed, alongside production of extracts with higher carvacrol content. Other components in the extracts present in significantly lower concentrations are the following: p-cymen, borneol, trans-caryophyllene, caryophyllene-oxide, γ-terpinene and linalool.In order to recognize the possibility of improving the extraction yield produced by employing supercritical carbon dioxide and enhancing the quality of extracts, the influence of several different pre-treatments was investigated. It was determined that the water pre-treatment raises the extraction yield the most (for 25%), while the ultrasonic pre-treatment was the most adequate choice for acquiring extracts with the highest content of carvacrol (66.46%). However, the highest carvacrol extraction yield from 100 g of dry weight was achieved by applying ethanol and water as pre-treatment.Extracts which were attained with subcritical water (by varying the temperature and time of extraction) were chemically characterized and Response Surface Method (RSM) was applied for the purposes of determining conditions of extractions under which the highest quality of extracts in terms of content of polyphenolic compounds and antioxidant activity is reached. The advantage of employing subcritical water over the classical method of extraction in respect of content of polyphenolic components and antioxidant activity was demonstrated.Dry extracts of S. montana obtained by using the spray drying technology (with different concentrations of maltodextrine) were examined in terms of their physical and chemical characteristics. Also, in order to investigate the possibility of their use as functional or food products, sensory analysis of attained extracts was performed, and furthermore, their pharmacological activity in terms of the ACE inhibitory and antioxidant effect was analysed as well.It was determined that the lowest concentration of maltodextrin (10%) was the most adequate for obtaining dry extracts of S. montana with the richest content of polyphenolic components and essential oil, and in respect of antioxidant and ACE inhibitory activities. Also, dry extract with 10% of maltodextrin meets all other physical-chemical parameters, which provide efficient drying and quality of extracts.In vivo activity of selected extracts of S. montana was examined on a model of hepatotoxicity induced by applying carbon tetrachloride. The influence of extracts on liver function, enzyme parameters of oxidative stress, and serum parameters of oxidative damage caused by carbon tetrachloride was analysed and hepatoprotective effect of S. montana extracts was determined. In addition, the influence of extracts on the growth of Ehrlich ascite tumors implanted in mice was investigated, and their impact on biochemical parameters of oxidative stress in tumor cells as well, and it was determined that the time of application of extracts is of great significance.

Tehnologije s superkritičnimi fluidi omogočajo pridobivanje produktov, z lastnostmi, ki jih je s klasičnimi postopki zelo težko ali celo nemogoče doseči. Visokotlačni procesi, kot je na primer postopek superkritične ekstrakcije, predstavljajo alternativo za izolacijo snovi iz G. lucidum z visoko biološko aktivnostjo. V prvem delu doktorske disertacije smo s pomočjo klasične ter superkritične ekstrakcije iz medicinske gobe G. lucidum skušali izolirati komponente z biološko aktivnostjo. Ekstrakcije smo izvedli iz zasnov trosnjakov t.i. primordijev ter iz zrelih trosnjakov gobe G. lucidum. Osredotočili smo se na »in vitro« biološke aktivnosti ekstraktov. Ekstraktom smo določili antioksidativno aktivnost po dveh različnih metodah. Prav tako smo določali še celokupne fenole, celokupne polisaharide ter proteine. Določili smo tudi inhibicijo encima acetilholinstearaze (AChE). Ekstraktom, ki smo jih pridobili s pomočjo superkritične ekstrakcije s CO2 smo določili »in vitro« protitumorsko aktivnost na HUVEC (Human Umbilical Vein Endothelial Cells - endotelijske celice iz popkovnične vene) ter na CaCo-2 (Caucasian colon adenocarcinoma - rakave celice debelega črevesa) celičnih linijah. V drugem pa smo se osredotočili na raziskovanje faznih ravnotežij β-karotena v subkritičnih in superkritičnih fluidih. V primeru ekstrakcije karotenoidov je potrebno le te iz rastlinskega tkiva hitro izolirati, da se izognemo oksidativni in encimski degradaciji. Glavni predstavnik karotenoidov je β-karoten. Nahaja se v številnih rastlinskih in živalskih tkivih, zato se ga da izolirati iz tkiv tudi s pomočjo uporabe SCF. Za načrtovanje visokotlačnih kemijskih procesov, kot je na primer ekstrakcija s SCF, je poznavanje in hkrati tudi razumevanje faznih ravnotežij ključnega pomena. Naš namen je bil v prvi vrsti določiti ravnotežne topnosti čistega β-karotena v sub- in superkritičnih fluidih (CO2 ter propanu). V nadaljevanju pa smo raziskovali fazna ravnotežja trikomponentnega sistema β-karotena – triolein – SC-CO2 v določenem tlačnem območju in pri različnih temperaturah. Dobljene rezultate o topnosti smo korelirali s Chrastilovim topnostnim modelom. Nowadays the technology of supercritical fluids allows producing products, that is very difficult or almost impossible to achieve by the conventional methods. High-pressure processes such as the process of supercritical extraction represent an alternative to the isolation of substances from the G. lucidum with the high biological activity. In the first part of the dissertation the conventional and supercritical extraction of the medical mushroom Ganoderma lucidum are presented. The biological active compounds from G. lucidum were thus isolated. Extractions were performed from immature fruiting body, so-called primordia, and from mature fruiting body. The biological activity of the extracts was determined "in vitro". The antioxidant activity was determined by the free-radical method and the antioxidant values of Trolox and ascorbic acid were obtained by Photochem method. The amount of total phenols, total polysaccharides, proteins and inhibition of antiacetycholinesterase (AChE) were also investigated. The cytotoxic activity of G. lucidum extracts obtained by supercritical extraction with CO2 was tested on HUVEC (human umbilical vein endothelial) and CaCo-2 (human colon adenocarcinoma) cell lines. In the second part of the dissertation the phase equilibria of β-carotene in sub- and supercritical fluids is reported. To avoid the oxidative and enzymatic degradation of carotenoids during the extraction process it is necessary to isolate them as fast as possible from the plant tissue. One of the most common carotenoids is β-carotene. It is located in a number of plant and animal tissues and can be isolated from the tissues by using supercritical fluids as solvent. Solubility or phase equilibrium data are very important for designing new high-pressure processes where supercritical fluids are used as solvents. The work presented here focused primarily on the equilibrium solubility of pure β-carotene in supercritical CO2 and subcritical propane. Additionally, the phase equilibrium of the ternary system of β-carotene – triolein – supercritical CO2 at different pressures and temperatures was investigated. Afterwards the obtained solubility data was correlated with the density based model purposed by Chrastil.

Cilj ovog rada je ukazivanje na mogućnost primene superkritične ekstrakcije sa ugljendioksidom na dekontaminaciju zemljišta zagađenih pesticidima (u ovom slučaju atrazinom). Analizirani rezultati ukazuju da se ova metoda veoma uspešno može koristiti za ostvarivanje istaknutog cilja. The aim of this paper is to study the applicability of supercritical extraction with carbon dioxide to the decontamination of soils containing pesticides (in this case with atrazine). The analyzed results indicate that this method can be successfully used for the realization of prominent purpose.

Konvencionalna ekstrakcija je še vedno primarna metoda, ki jo uporabljamo za ekstrahiranje olja iz različnih semen, medtem, ko je pri tej diplomski nalogi bila izbrana alternativna metoda konvencionalni metodi, to je ekstrakcija z uporabo superkritičnih in subkritičnih fluidov. Uporaba superkritične ekstrakcije (SCE) ima kar nekaj pozitivnih lastnosti, med drugim tudi to, da končni produkti ne vsebujejo ostankov topil. Laboratorijsko delo je bilo razdeljeno na tri dele. Prvi del je obsegal ekstrahiranje bučnega olja sorte – Gleisdorfska golica z uporabo visokotlačne ekstrakcijske naprave, pri čemer smo kot topilo uporabljali superkritični ogljikov dioksid ter subkritični propan. Z namenom preučevanja kinetike ekstrakcije, smo pri obeh topilih spreminjali temperaturo in tlak, ter tako ugotavljali kako ta dva parametra vplivata na hitrost in izkoristek ekstrakcije. V drugem delu eksperimenta smo s pomočjo saponifikacije hidrolizirali maščobe in pri tem dobili soli maščobnih kislin, ki smo jih nato v tretjem delu laboratorijskega eksperimenta analizirali z uporabo plinske kromatografije (GC) in tako določili vsebnost prostih maščobnih kislin v ekstraktih. Na podlagi vseh opravljenih meritev smo na koncu lahko določili optimalne pogoje ekstrahiranja in izbrali primerno topilo, ki je dalo največje izkoristke. Rezultati so pokazali, da se olje bučnih semen veliko bolje topi v propanu, kar smo opazili tudi med samim eksperimentom na podlagi barve ekstrakta, ki je v primeru uporabe superkritičnega ogljikovega dioksida bila rjavo – rumena, medtem ko pri ekstrakciji s subkritičnim propanom temno zelena. Izkoristki ekstrakcije so bili nad 30 %, vendar pri propanu v povprečju nekoliko višji (do 37 %) v primerjavi z ogljikovim dioksidom (do 34 %). Izkazalo se je tudi, da izkoristki ekstrakcije naraščajo z višanjem temperature in večanjem tlaka. Conventional extraction is still the primary method used for extracting oil from various seeds, however, in this thesis we decided to use the alternative method – extraction with supercritical and subcritical fluids. Supercritical extraction (SCE) has various positive properties, for example, the final products do not contain traces of the solvent. The laboratory work was divided into three major parts. The first part covered the extraction of variety of pumpkin oil – Gleisdorfska golica using the laboratory high pressure extraction device, where supercritical carbon dioxide and subcritical propane were used as solvents. During the experiment I was changing the temperature and pressure with the aim of studying the extraction kinetics and to identify how these parameters influence the rate and yield of extraction. Saponification was used in the second part of the experiment in order to hydrolyze fats into fatty acid salts. These fatty acid salts were then analyzed in the third part of the experiment, using gas chromatography. As a result I was able to determine the content of free fatty acids in my extracts. I was able to determine the optimal conditions of the extraction and choose suitable solvent, which gave the best extraction yield based on all measurements. Results are showing that oil is more soluble in subcritical propane than in supercritical carbon dioxide, which was also visible during the experiment based on the color. When using the supercritical carbon dioxide, the color of extract was brown to yellow, on the other side, when using the subcritical propane, the color was dark green. The extraction yields were above 30 % , though in the case of propane they were slightly higher on average (up to 37 %) than in the case of carbon dioxide (up to 34 %). It turned out, that extraction yields are increasing when both, temperature and pressure, are increasing.

Namen diplomske naloge je bil z različnimi ekstrakcijskimi postopki iz štirih vrst semen (sončnično, laneno, sezamovo in konopljino seme) pridobiti olja ter določiti vsebnost trigliceridov (TG) in njihove antioksidativne aktivnosti. Ekstrakcijo olj smo izvedli s štirimi različnimi ekstrakcijskimi postopki, in sicer konvencionalno z mešanjem v topilu in na Soxhlet aparatu, kjer smo kot topilo uporabili heksan ter s semikontinuirano superkritično ekstrakcijsko napravo, kjer smo kot topilo uporabili ogljikov dioksid in propan. Ekstrakcije s superkritičnim fluidom CO2 in subkritičnim fluidom propanom smo izvedli pri dveh temperaturah: 40 ºC in 60 ºC, ter tlaku 300 bar. Za posamezno seme smo določili, katera ekstrakcijska metoda prinaša največji ekstrakcijski izkoristek olja. Vsebnost TG v ekstrahiranih oljih smo določili s tekočinsko kromatografijo visoke ločljivosti (HPLC) –ELSD, antioksidativno kapaciteto za olje topne vzorce pa smo določili s foto-luminiscenčno (Photochem) metodo. Dobljeni rezultati kažejo, da imajo največji ekstrakcijski izkoristek olja sončničnega semena (50-55 ut. %). Glede na izkoristke si nato sledijo sezamovo seme (45-50 ut. %), laneno seme (30-40 ut. %) in konopljino seme (25-30 ut. %). Ekstrahirana olja vsebujejo po večini nenasičene trigliceride. Sončnično in sezamovo olje vsebuje gliceril dilinoleat – oleat (LLO), gliceril trilinoleat (LLL), gliceril trioleat (OOO) in gliceril linoleat – dioleat (LOO), torej trigliceride, ki vsebujejo linolno in oleinsko kislino. V konopljinem olju se nahajajo naslednji trigliceridi: gliceril trilinoleat (LLL), gliceril oleat – dilinolenat (OLnLn) in gliceril dilinoleat – oleat (LLO). Ti trigliceridi vsebujejo največ linolne kisline. Največji delež TG v lanenem olju predstavljata gliceril trilinolenat (LnLnLn) in gliceril oleat – dilinolenat (OLnLn). Prevladuje torej linolenska kislina. Olja iz teh semen imajo dobre antioksidativne aktivnosti, kar nakazuje na njihovo visoko vsebnost oljetopnih antioksidantov. The aim of this thesis was the extraction of oils from different seeds (sunflower, flaxseed, sesame, hempseed) with four extraction techniques and determination of their triglyceride (TG) profile as well as measuring their antioxidant activity. For oil extraction four different extraction procedures have been used: conventional extraction procedures with mixing in hexane and Soxhlet extraction with hexane and supercritical fluid extraction with carbon dioxide and propane. The extractions with a supercritical fluid CO2 and subcritical fluid propane were done at two different temperatures: 40 °C and 60 °C, with a pressure of 300 bars. Comparisson of yield of oils obtained with applied methods were made. Triglyceride content in obtained oils was determined by high performance liquid chromatography (HPLC) -ELSD. The antioxidant activity of the oils was measured using photo-luminiscence (Photochem) method. The obtained results show that the maximum extraction efficiency of oil was obtained for sunflower seeds (50-55 wt. %) followed by sesame seeds (45-50 wt. %), flaxseeds (30-40 wt. %) and hempseeds (25-30 wt. %). Extracted oils contain mostly unsaturated triglycerides. Sunflower and sesame oil contain glyceryl dilinoleate - oleate (LLO), glyceryl trilinolenate (LLL), glyceryl trioleate (OOO) and glyceryl linoleate – dioleate (LOO), TG which contain linoleic and oleic acid. The hempseed oil triglycerides are glyceryl trilinoleate (LLL), glyceryl oleate – dilinolenate (OLnLn) and glyceryl dilinoleate – oleate (LLO), mostly TG of linoleic acid. The largest proportion of TG in flaxseed oil represents glyceryl trilinolenate (LnLnLn) and glyceryl oleate - dilinolenate (OLnLn) with the predominant linoleic fatty acid. Oils from these seeds have shown to have good antioxidant activity, which indicates a high content of oil-soluble antioxidants.

U okviru ove doktorske disertacije, proučavana je ekstrakcija bosiljka (Ocimum basilicum L.) primenom postupka hidrodestilacije, Soxhlet ekstrakcije i ugljendioksidom u superkritičnom stanju. Određen je sadržaj etarskog ulja u drogi (0,565%), a primenom GC/MS(FID) metoda određen kvalitativni i kvantitativni sastav detektovanih komponenata. Identifikovane komponente svrstane su u nekoliko grupa (monoterpeni, seskviterpeni i njihovi oksidovani derivati). Rezultati ispitivanja pokazuju da je dominantno jedinjenje etarskog ulja bosiljka linalool (50,09%), i da ono pripada linaloolskom hemotipu (hemotip A).Ispitan je uticaj temperature (40-60°C) i pritiska (100–300 bar) superkritičnog ugljendioksida na promenu prinosa ekstrakta. Utvrđeno je da se kumulativni prinos ekstrakta povećava sa porastom pritiska ekstrakcije, dok u zavisnosti od temperature, prinos ekstrakcije zavisi od gustine rastvarača i napona pare komponenata u ekstraktu.Primenom sukcesivne (frakcione) superkritične ekstrakcije izvršeno je frakcionisanje, a dobijeni ekstrakti upoređivani u pogledu prinosa, kvalitativnog i kvantitativnog sastava.Ispitivanja hemijskog sastava CO2 ekstrakta (GC/MS i GC/FID) pokazala su da ekstrakti pored komponenata etarskog ulja sadrže i druga lipofilna jedinjenja (voskove, masna ulja, masne kiseline, fitosterole i sl.). Dominantne komponente u svim ispitivanim ekstraktima su linalool, eugenol i δ-kadinen, dok su α-bergamoten, germakren D, γ-kadinen, β-selinen i spatulenol komponente sa manjim saržajem.Na osnovu eksperimentalnih rezultata kinetike superkritične ekstrakcije izvršeno je modelovanje ekstrakcionog sistema bosiljak – superkritični CO2, primenom tri modela (Kandiah i Spiro, Brunner i Esquivel i sar.). Na osnovu srednjih vrednosti relativnog odstupanja modelovanih i eksperimentalnih vrednosti prinosa ekstrakcije, utvrđeno je da model Kandiah i Spiro pokazuje prihvatljivo slaganje, dok ostali modeli pokazuju lošije slaganje eksperimentalno određenih i modelom izračunatih prinosa ekstrakta.U završnoj fazi rada ispitan je antioksidativni potencijal etarskog ulja i CO2 ekstrakata, primenom DPPH metode. Najveću antioksidativnu aktivnost pokazao je ekstrakt dobijen na 100 bar, 60°C (IC50 = 18,93 μg/ml), koja je 5 puta veća od antioksidativne aktivnosti etarskog ulja bosiljka., In this study, the extraction of basil (O. basilicum L.) ussing the hydrodistillation process, Soxhlet extraction and supercritical extraction with carbon dioxide were investigated. The content of essential oil was determined (0,565%), and the qualitative and quantitative composition of detected coumpound was made by GC/MS(FID). The identified coumpounds were classified into five groups (monoterpenes, sesquiterpenes and their oxidative derivates). The results of investigation shows that the dominant compound of basil essential oil is linalool (50,09%), and that it belongs to linalool chemotype (chemotype A).The influence of temperature (40-60°C) and pressure (100–300 bar) of supercritical carbon dioxide on extraction yields were studied. It was determined that the total yield of extract increases with the increaseing of extraction pressure, while the influence of temperature on the yield depends on the density of the solvent and vapor pressure of components in the extract.Fractional supercritical fluid extraction was performed, and obtained extracts were compared regarding of yield, and the qualitative and quantitative composition of the extract. Chemical analyses of CO2 extract by GC/MS and GC/FID showed that extract contains, besides essential oil componentes, other lipophilic compounds (waxes, fatty oils, fatty acids, phytosterols, etc.). The few compounds were dominant in all analyzed basil extracts: linalool, and to a lesser extent eugenol, δ-cadinene, α-bergamotene, germacrene D, γ-cadinene, β-selinene and spathulenol. Based on the experimental results of the extraction kinetics, a modeling of basil – supercritical CO2 extraction system was performed, using three models (Kandiah and Spiro, Brunner, and Esquivel et al.). Based on the average values of the relative deviations of modeled and experimental values of extraction yield, it was found that the model Kandiah and Spiro shows acceptable agreement, while other models show lower agreement of the experimentally determined and model calculated extraction yield.In finale phase of work the antioxidant potential of essential oil and CO2 extracts were examined, using DPPH method. The highest antioxidant activity showed CO2 extract obtained at 100 bar, 60°C (IC50 = 18,93 μg/ml), which is 5 time higher then antioxidant activity of basil essential oil.

V diplomski nalogi smo s konvencionalno in superkritično ekstrakcijo ekstrahirali zmleto mandarinino lupino. Mandarinine lupine predstavljajo pomemben vir hesperidina in polimetoksi flavonov, kot sta nobiletin in tangeretin. Izvajali smo naslednje metode ekstrakcije: - konvencionalna ekstrakcija z 70 % acetonom - superkritično ekstrakcijo s CO2. Superkritične ekstrakcije smo opravili z uporabo laboratorijske ekstrakcijsko visokotlačne naprave, pri kateri smo s črpalko črpali CO2 iz jeklenke v avtoklav skozi zdrobljene mandarinine lupine (suhe, sveže) pri tlakih (100, 200, 300 bar) in temperaturah (40, 60, 80 °C). Ekstrakte (mandarinino olje) smo shranili v embalažah,ki so bile zaščitene pred svetlobo v zamrzovalniku. Največje izkoristke pri postopku superkritičnih ekstrakcij z ogljikovim dioksidom smo dobili pri vlažnih mandarininih olupkih pri tlaku 300 bar in temperaturi 80 °C. Izkoristek je znašal 1,36 %. S pomočjo eksperimentalnega načrta po Taguchi-ju smo določili optimalne pogoje separacije ter vpliv procesnih parametrov. Za analizo ekstraktov s HPLC metodo pri tem smo ekstrakte pridobljene s konvencionalno ekstrakcijo raztopili v 100 % metanolu z namenom, da določimo sestavo in vsebnost flavonoidov, predvsem hesperidina v ekstraktih. Izvedene so bile raziskave na področju predobdelave materiala s superkritičnimi fluidi: določili smo vpliv predobdelave materiala na izolacijo hesperidina iz mandarininih olupkov. In our diploma thesis mandarin peels have been extracted using conventional and supercriticial extraction methods. Mandarin peels present an important source of hesperidin and polimetoxy flavones. Extraction methods used are - conventional extraction with 70 % acetone - supercritical extraction with CO2. Supercritical extractions were carried out using the laboratory high-pressure extraction plant, which pumps carbon dioxide and propane from cylinder thrugh the mandarin peels in the extractor at pressures (100, 200, 300 bar) and temperatures (40, 60, 80 °C). Extracts (mandarin peels) have been stored in containers protected from light in the freezer. Maximum yields of supercritical extraction with carbon dioxide were obtained with fresh mandarin peels at pressure 300 bar and temperature 80 °C. Yield was 1,36 %. With Taguchi experiment design we established optimal conditions for separation and influence of process parameters. For the analysis of extract with HPLC procedure are extracts of obtained with conventional extraction dissolved in 100 % methanol with intention, that we determine composition and content of flavonoid, above all hesperidin in extracts. Pre-treatment of mandarin peels were performed with supercritical fluid: we established the influence of pre-treatment of material on the isolation of hesperidin from mandarin peels.

Pridobivali smo sezamovo olje iz sezamovega semena. Uporabili smo dva postopka ekstrakcije – konvencionalno ekstrakcijo z različnimi topili in superkritično ekstrakcijo. Kot topila pri konvencionalni ekstrakciji smo uporabili heksan, etanol in petroleter. Pri superkritični ekstrakciji smo kot topilo uporabili propan. Imeli smo šest različnih ekstrakcijskih pogojev. Na podlagi dobljenih rezultatov in dobljenega olja smo izračunali izkoristke ekstrakcij. Vsak ekstrakt smo tudi umilili. Vzorcem smo določali vsebnost maščobnih kislin v olju in antioksidativno kapaciteto olja. Vsebnost maščobnih kislin smo določali s plinsko kromatografijo in ugotovili, da prevladujeta oleinska in linolna maščobna kislina. Antioksidativnost smo določali po metodi Photochem. Največji izkoristek pri konvenkcionalni ekstrakciji smo dobili s topilom heksan, pri superkritični ekstrakciji pa pri pogojih T=50°C in p=100bar. Ugotovili smo, da je sezam dobra oljna rastlina, saj smo imeli čez 50% izkoristke ekstrakcije. Glede na rezultate in potek eksperimentov smo ugotovili, da je boljša superkritična ekstrakcija, kot pa klasična konvencionalna ekstrakcija, saj je izkoristek bistveno večji, pa tudi olje je čistejše. The main purpose of this work is extraction of sesame oil from sesame seeds and extracts analysis. Extraction was performed using the two different methods - conventional extraction with different solvents and supercritical extraction. Solvents that were used for conventional extraction are hexane, ethanol and petroleum ether. Solvent that was used in supercritical extraction is propane. In this research we considered six different conditions of extraction. Based on the results and obtained oil we also calculated the efficiency of extraction. All extracts were saponified and fatty acids present in each sample were analysed by gas chromatography. Results showed, that in samples oleic and linoleic fatty acid dominated. Antioxidant capacity was determined by Photochem method. Maximum efficiency with conventional extraction method, was achieved by solvent hexane, while with supercritical extraction, that was performed at the following conditions: T=50°C and p=100bar. The results of the study suggest that sesame is a good oil plant, because efficiency of extraction were above 50%. Based on the results it can be concluded that between both methods of extraction used, the supercritical extraction is better than conventional. Efficiency was significantly higher and also the oil obtained was cleaner.

Primerjali smo učinkovitost superkritične in konvencionalne metode ekstrakcije antioksidativnih komponent iz kurkume (Curcuma longa). Superkritično ekstrakcijo smo izvajali s pomočjo laboratorijske visokotlačne ekstrakcijske naprave, kjer smo s črpalko črpali ogljikov dioksid iz jeklenke v avtoklav skozi material pri različnih pogojih tlaka in temperature. Konvencionalne ekstrakcije smo opravljali po naslednjih metodah: ekstrakcija s topili pri povišani tlakih, ekstrakcija po Soxhletu, ekstrakcija s hladnim topilom v bučki in ekstrakcija z ultrazvokom. Kot topilo smo uporabljali absolutni etanol pri vseh metodah konvencionalne ekstrakcije, medtem ko smo topilo heksan uporabljali pri ekstrakciji po Soxhletu in pri ekstrakciji s hladnim topilom v bučki. Dobljene ekstrakte smo shranjevali v zamrzovalniku, zaradi občutljivosti na svetlobo. V dobljenih ekstraktih smo določali vsebnost antioksidativnih komponent. Izkoristki superkritične ekstrakcije so bili približno enaki pri vseh delovnih pogojih. Kot smo lahko videli, pri superkritični ekstrakciji z ogljikovim dioksidom na topnost topljenca minimalno vplivata tlak in temperatura, saj smo pri najvišjem tlaku in temperaturi dobili največji izkoristek. Pri konvencionalni ekstrakciji po Soxhletu in pri ekstrakciji s hladnim topilom v bučki je bilo olje bolj topno v heksanu, ki je nepolarno topilo kot v etanolu. Antioksidativne komponente smo določali po naslednjih metodah: z radikalsko metodo – DPPH, z določitvijo v lipidih topnih antioksidativnih komponent s Photochemom, z beta-karotensko metodo in z metodo celokupnih fenolov. Ugotovili smo, da imajo najmanj antioksidativnih komponent ekstrakti pridobljeni s topilom heksan. Določali smo tudi prisotnost določenih kurkuminoidov v ekstraktih s tankoplastno kromatografijo. Videli smo, da v ekstraktih pridobljenih s heksanom ni bilo prisotnih kurkuminoidov. Določali smo tudi antimikrobno aktivnost na bakteriji Bacillus cereus in Escherichia coli. Ugotovljeno je bilo, da povzročijo največjo inhibicijo rasti ekstrakti pridobljeni s topilom etanol pri povišanih tlakih. The comparison between supercritical and conventional extraction of antioxidative compounds from Turmeric (Curcuma longa) has been performed. The supercritical extraction was performed with the laboratory high presure extraction device where the carbon dioxide was pumped from the tank to the extractor (through the material) at different conditions of pressure and temperature. The conventional extraction methods used were: extraction with solvents at high presures, Soxhlet extraction, extraction with cold solvents in a flask and extraction with ultrasound. As a solvent absolute ethanol was used in all methods of conventional extraction, while hexane was used in the Soxhlet extraction and extraction with cold solvent in a flask. Due to their sensitivity to sunlight, the obtained extracts were preserved within a freezer. The obtained extracts were analyzed on the content of antioksidative components. The efficiency of the supercritical extractions were very close at all working conditions. Temperature and pressure have minimal impact on the extraction yield the highest yield was obtained at the highest pressure and highest temperature. With the conventional extraction with Soxhlet and with the cold solvent extraction in a flask the oil was more soluble in hexane which is a non-polar solvent. Antioxidative components were analyzed by following methods: radikal method - DPPH, determination of antioxidative capacity of the lipid soluble compounds (ACL) with Photchem, beta-carotene method and with the method of total fenols. The lowest amount of antioxidative components were extracted with hexane. Furthermore, the presence of curcuminoids in extracts was determined with thin layer cromotagraphy. It was observed that extracts gained with hexane contained no curcuminoids. The antibacterial activites of extracts on the bacteria Bacillus cereus and Escherichia coli were also investigated. The largest growth inhibition was observed for extracts obtained with extraction with absolute ethanol at high presures.

Za določanje vsebnosti vanilina v zmletih strokih vanilije smo uporabili tri vrste ekstrakcij, in sicer: ekstrakcijo s superkritičnima fluidoma argonom in CO2 pri temperaturah 25 °C, 40 °C in 60 °C ter tlakih 150 bar in 300 bar, ekstrakcijo po Soxhletu in konvencionalno ekstrakcijo, obe s topiloma metanol in etanol. Rezultati kažejo, da dobimo z ekstrakcijo po Soxhletu in konvencionalno ekstakcijo zelo velike izkoristke, tudi nad 50 %, in da so le-ti večji pri topilu metanol, kar pomeni, da je ta primernejše topilo od etanola. Drugače kažejo rezultati pri ekstrakciji s superkritičnimi fluidi, kjer so izkoristki zelo nizki, in sicer okrog 1 % pri argonu in 5 % pri CO2. Dobljene ekstrakte smo raztopili v metanolu in jim merili absorbance pri valovnih dolžinah 228,5 nm (vanilin) in 270 nm (OEV). UV analize so pokazale, da je vsebnost vanilina v ekstraktih, pridobljenih z ekstrakcijo s superkritičnimi fluidi, višja kot v ekstraktih, pridobljenih z ekstrakcijo po Soxhletu ali s konvencionalno ekstrakcijo. Iz tega lahko sklepamo, da so superkritični fluidi veliko bolj selektivna topila. Določali smo topnost vanilina in orto etilvanilina v argonu pri temperaturah 40 °C, 60 °C in 80 °C ter tlakih v območju do 500 bar ter rezultate primerjali s topnostjo vanilina in OEV v CO2. Ugotovili smo, da je pri nižjih temperaturah in tlakih do približno 200 bar topnost višja v argonu. Z dodatnim višanjem tlaka topnost narašča počasneje. Three types of extraction were used to determine the content of vanillines in milled vanilla pods: extraction with supercritical fluids argon and CO2 at temperatures of 25 °C, 40 °C and 60 °C and at pressures of 150 bar and 300 bar, Soxhlet extraction and conventional extraction, both with solvents methanol and ethanol. Very high yields appear in the case of Soxhlet and also conventional extraction, even higher than 50 %, furtheremore they higher when using methanol as a solvent. Therefore methanol resulted as a more adequate solvent than ethanol. Yields are low in case of extraction with supercritical fluids approximately 1 % with argon and 5 % with CO2. Obtained extracts were dissolved in methanol and absorbance was measured at a wavelength of 228,5 nm (vanillin) and at a wavelenght of 270 nm (OEV). UV analysis show higher content of vanillin in extracts, obtained by extraction with supercritical fluids (supercritical extraction), than in those obtained with the Soxhlet extraction or conventional extraction. Therefore it can be concluded that supercritical fluids are way more selective than conventional solvents. Solubility of vanillin and ortho ethylvanillin in argone at temperatures of 40 °C, 60 °C and 80 °C and at pressures up to 500 bar was also determined. Results were compared to solubility in of OEV and CO2. It was clearly observed that solubility is higher in argon under lower temperatures and pressures up to about 200 bar. With additional increase of pressure only a slight impact on solubility is noted.

V diplomski nalogi smo izvedli ekstrakcijo olja iz semen industrijske konoplje z zgoščenimi plini in konvencionalnimi topili. Namen diplomske naloge je bil raziskati vpliv različnih ekstrakcijskih tehnik na izkoristek ekstrakcije ter vsebnosti maščobnih kislin v konopljinem olju. Superkritične ekstrakcije smo izvedli z zgoščenima plinoma propan ter ogljikov dioksid pri konstantnih temperaturah (50 in 70 °C) ter konstantnih tlakih (200, 300, 480 bar). Ekstrakcije s konvencionalnimi topili smo izvedli po metodi ekstrakcije po Soxhletu (z etanolom, heksanom in petroletrom), ekstrakciji z etanolom pri povišanih tlakih (100 in 200 bar) ter ultrazvokom. Raziskali smo vsebnost maščobnih kislin v konopljinem olju. Analizo vzorcev smo opravili s plinsko kromatografijo. In our diploma thesis we performed an extraction of hempseed oil from the seeds of industrial hemp with dense gases and conventional solvents. The intention of the diploma thesis was to research the influence of different extraction techniques on the amount of extraction and on the fatty acids content in hempseed oil. Supercritical extractions were carried out with dense gases propane and carbon dioxide at constant temperatures (50 and 70°C) and at constant pressures (200, 300, 480 bar). Extractions with conventional solvents were carried out according to extraction method with Soxhlet extraction (with ethanol, hexane and petroleum ether), extractions with ethanol were carried out at increased pressures (100 and 200 bar) and with ultrasound. We researched the fatty acids content in hempseed oil. Sample analysis was carried out with gas chromatography.

Upotrebom različitih ekstragenasa izvršena je ekstrakcija ploda, lista i korena belog (Morus alba L.) i crnog duda (Morus nigra L. ). Ekstrakcija etanolom je izvršena pri predhodno određenim optimalnim uslovima ekstrakcije (70% koncentracija etanola, temperatura 60°C i odnos rastvarač:droga 20 ml/g). Ispitan je hemijski sastav ekstrakata različitih delova duda. Ispitivanja hemijskog sastava etanolnih ekstrakata obuhvatala su određivanje sadržaja ukupnih fenola, ukupnih flavonoida, ukupnih i monomernih antocijana, kvalitativni i kvantitativni sastav polifenolnih komponenata, kao i sadržaj mikro-, makro- i toksičnih elemenata. Najvedi sadržaj ukupnih fenolnih jedinjenja određen je u ekstraktu korena M. nigra (186 mg EHK/g), a najvedi sadržaj flavonoidnih jedinjenja u ekstraktu lista M. nigra (67 mg ER/g). U ekstraktima lista duda po svom sadržaju u odnosu na ukupan sadržaj polifenolnih jedinjenja ističe se kafena kiselina (od 53,34 do 62,11%), dok se u ekstraktima korena izdvaja hlorogenska kiselina (udeo vedi od 74%). U pogledu mineralnog sastava ekstrakata relativno veliki sadržaj Ca i Mg je karakterističan za obe vrste ekstrakata (vedi od 1.400 μg/g). Mikroelementi u svim ispitanim uzorcima duda po opadajudem nivou, su Fe, B, Zn, Cu i Mn. U ekstraktima dobijenim superkritičnim CO2 određen je sadržaj masnih kiselina i ukupnih karotenoida. U ispitivanim ekstraktima dominante su nezasidene masne kiseline, linolna i linolenska kiselina, a značajan je udeo i zasidene masne kiseline (palmitinske kiseline). Ispitivanje delovanja ekstrakata obuhvatalo je evaluaciju antioksidativne aktivnosti pradenjem sposobnosti neutralizacije slobodnih radikala, redukcionog potencijala i inhibicije lipidne peroksidacije. Po svim primenjenim metodama najvedu aktivnost je pokazao ekstrakt lista crnog duda. U završnoj fazi rada ispitana je in vitro citotoksična aktivnost ekstrakata duda, njihovim delovanjem na rast tri delijske linije: Hep2, RD i L2OB. Ekstrakti lista M. nigra su se pokazali kao najpotentniji inhibitori rasta na sve tri delijske linije (IC50 B> Zn> Cu> Mn. In the extracts obtained by supercritical CO2 fatty acids and total carotenoids were determined. In these extracts dominant unsaturated fatty acids were linoleic and linolenic acid. Also, significant proportion of saturated fatty acid (palmitic acid) has been detected. Testing of extracts activity included the evaluation of antioxidant activity applying the methods based on: neutralize free radicals, reducing power potential and the inhibition of lipid peroxidation. All applied methods showed the highest activity of black mulberry leaf extract. In the final phase of work in vitro cytotoxic activity of mulberry extracts was examined. Their effect on growth of three cell lines: Hep2, RD, and L2OB were analysed. Extracts of M. nigra proved to be the most potent inhibitors of growth in all three cell lines (IC50

Upotrebom 50% etanola i superkritičnog ugljendioksida kao ekstragensa izvršena je ekstrakcija različitih vrsta pečuraka. Nakon određivanja prinosa suvih ekstrakata, u njima su određeni: sadržaj ukupnih fenola i ukupnih flavonoida, sadržaj makro- i mikro-elemenata, sadržaj adenozina, kao i sadržaj karakterističnih komponenata pistilarina i variegetične kiseline. Najveći sadržaj ukupnih fenolnih jedinjenja određen je u suvom ekstraktu pečurke C. pistillaris, najveći sadržaj ukupnih flavonoida u ekstraktu D. confragosa, a najviše esencijalnog cinka i selena sadrži suvi ekstrakt pečurke B. edulis. U odnosu na ostale suve ekstrakte ekstrakt pečurke L. saccatum sadrži nekoliko puta veću količinu adenozina. Antioksidativno delovanje suvih ekstrakata analizirano je primenom spektrofotometrijskih i EPR metode. Ekstrakti C. pistillaris, B. edulis i A. mellea pokazali su se kao najefikasniji skevindžeri najopasnije slobodnoradikalske vrste hidroksil radikala, dok se efikasnim u prevenciji lipidne peroksidacije mogu smatrati ekstrakti B. edulis, A. mellea i L. saccatum. Nakon određivanja prinosa supekritičnih ekstrakata ispitivanih pečuraka, u njima je analiziran sastav masnih kiselina i detektovana jedinjenja sterolnog tipa. U ispitivanim ekstraktima dominantna je nezasićena linoleinska kiselina, a značajan je udeo i oleinske nezasićene masne kiseline. Dominantno jedinjenje sterolne strukture u gotovo svim ekstraktima je ergosterol. Ispitan je uticaj superkritičnih ekstrakata pečuraka na fluidnost membrane eritrocita i na osnovu dobijenih rezultata zaključeno da bi značajnu ulogu u antihipertenzivnoj ishrani mogle imati pečurke A. mellea i M. procera., Using 50% ethanol and supercritical carbon dioxide different mushroom extracts were prepared. After analysis of extraction yield, content of total phenols and total flavonoids, content of macro-elements and micro-elements, as well as content of adenosine and characteristic compounds (pistilarin and variegatic acid) were determined. Highest content od total phenols was determined in C. pistillaris extract, highest content of total flavonoids in D. confragosa and highest content of essential zinc and selenium in B. edulis dry extract. In comparsion to other extracts L. saccatum posses few time higher content of adenosine. Antioxidat activity was analysed by spectrophotometric and EPR methods. Extracts of C. pistillaris, B. edulis and A. mellea have been showed as a most efficient in scavenging of dangerous OH˙ radical. In lipid peroxidation prevention significant were mushrooms extracts of B. edulis, A. mellea and L. saccatum.After determination of mushrooms supercritical extraction yield, fatty acid composition and sterol components were analysed. Dominant unsaturated fatty acid in investigated mushroom extracts was linoleic acid. Content of oleic fatty acid was also significant. Dominant compound of sterol structure, in almost all supercritical extracts, was ergosterole. Influence of supercritical mushroom extracts on eritrocite membrane fluidity was investigated. Acording to obtained results, mushrooms A. mellea and M. procera could have significant role in antihypertensive diet.

U radu je ispitivan utjecaj temperature ekstrakcije (40 – 50 °C), tlaka ekstrakcije (100 – 300 bar) i vremena ekstrakcije (2 – 4 h) kao i njihove interakcije na ekstraktibilnost ulja iz usitnjenog zrna soje sorte „Ika“, srednjeg promjera čestica 0,3131±0,01 mm, pomoću superkritičnog CO2. Volumni protok superkritičnog CO2, za provedeni opseg istraživanja, bio je konstantan i iznosio je 1,629 L/min. Metodom odzivnih površina, pomoću Box-Behnken-ovog dizajna, određeni su optimalni uvjeti ekstrakcije (temperatura: 50 °C, tlak: 300 bara i vrijeme: 4 h); za istraživani raspon eksperimentalnih podataka, pri kojima je modelom predviđen maksimalni prinos ulja od 6,55 g ulja/100 g usitnjenog zrna soje. Analiza varijance pokazala je da postoji značajan utjecaj tlaka i vremena ekstrakcije, kao i njihove interakcije, na prinos ulja. Temperatura ekstrakcije nije značajno utjecala na prinos ulja, niti promatrana kao pojedinačan parametar, niti prilikom interakcije s drugim ispitivanim parametrima. Za opisivanje kinetike superkritične CO2 ekstrakcije ulja iz zrna soje uspješno je korišten jednofazni difuzijski model koji je pokazao dobra slaganja s eksperimentalnim podacima. The effects of temperature (40 – 50 °C), pressure (100 – 300 bar) and extraction time (2 – 4 h) and their interactions on the extractability of oil from milled soybeans variety „Ika“ mean particles size diameter 0.3131±0.01 mm using supercritical CO2 were studied. Volumetric solvent flow rate was kept constant at 1.629 L/min. The optimal conditions for the extraction of oil (temperature: 50 °C, pressure: 300 bar and time: 4 h) were determined by response surface methodology using the Box-Behnken design. At the above menitioned conditions, maximum oil yield was 6.55 g oil/100 g milled soybeans. The analysis of variance showed that the effect of pressure and extraction time, as well as their interaction, on the oil yield was significant. Extraction temperature had no significant effect on the oil yield, nor as a single parameter, nor in interaction with other examined parameters. One stage diffusion model has been successfully used to describe the supercritical CO2 extraction kinetics of oil from soybeans. The experimental values agreed well with those predicted by diffusion model.

Pyrethrum (Chrysanthemum cinerarifolium Vis.) is endemic plant to Montenegro, Croatia and Albania. Insecticidal pyrethrin compounds (pyirethrin I i II) extracted from pyrethrum flowers. As pyrethrum research has been scarce in this country so far our primary objective was to provide data on the morphology and type of habitats of wild pyrethrum ina varius parts of Montenegro and data on its introduction into climate conditions of central Serbia. The following morphological and productivity parameters were investigated: Cluster width, cluster height, number of shoots and flower - bearing stems, number of flower-heads, fresh and dry weight of roots, stem, leaves and flowers. Determinate and seed germination and pyrethrum content. The results show the pyrethryn mean in the flower be around 0,7% while most favorable properties were found in plant of the Trsteno locality. Percent of pyrethrin was simual in the plants growed in ecological conditions of central Serbia. Buhač (Chrysanthemum cinerarifolium Vis.) je endemična biljka rasprostranjena u Crnoj Gori, Hrvatskoj i Albaniji. Iz cvasti buhača se ekstrahuju insekticidna jedinjenja piretrini (piretrin I i II). U našoj zemlji ispitivanja buhača su bila veoma oskudna i zato je cilj ovog rada bio dobijanje podataka o biološkim osobinama samoniklog buhača prikupljenog sa različitih staništa Crne Gore kao i mogućnost njegove introdukcije u uslove centralne Srbije. Ispitivani su sledeći morfološki i produktivni parametri: širina i visina bokora, broj izdanaka i cvetonosnih stabala, broj glavica cvasti, sveža i suva masa korena, stabla, listova i cvasti. Ispitivana je i klijavost semena buhača i određivan je procenat ukupnih piretrina. Dobijeni rezultati pokazuju da najpovoljnije osobine od svih ispitivanih populacije imaju biljke sa lokaliteta Trsteno. Procenat piretrina u cvastima buhača sa ispitivanih lokacija je oko 0,7 %, a takođe, može se zaključiti da se gajenjem u ekološkim uslovima Srbije procenat piretrina nije značajno smanjio.