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Veliki dio aromatskih spojeva nalazi se u vinu u vezanom obliku najčešće vezano na šećernu jedinicu. Kako bi se vinu povećao aromatski intenzitet potrebno je te vezane aromatske spojeve osloboditi. To se kidanje veze obavlja uz pomoć enzima, točnije enzima iz skupine glikozidaza – β-glukozidazama. Takvo se oslobađanje najčešće vrši na aromatičnim sortama kako bi se njihov intenzitet još više izrazio i postao primamljiviji krajnjim potrošačima. U ovom radu istraživana su dva enzima, Lallzyme Beta i Endoyzm β-Split na vinima 'Muškata žutog' i 'Muškat ruže'. Nakon tretmana provedene kemijske analize pokazale su pozitivan utjecaj korištenih enzima na povećanje koncentracije aromatskih spojeva. Posebno se pozitivan utjecaj pokazao kod spojeva poput terpena i estera koji su značajno povećali voćni i cvjetni intenzitet vina. Na kraju, senzorna ocjena ukazala je na različitost tretiranih vina te se može zaključiti kako je korištenje ovih enzima opravdano sa ciljem proizvodnje vina veće kakvoće. A large part of aromatic compounds is found in wine in a bound form, usually bound to a sugar unit. In order to increase the aromatic intensity of the wine, it is necessary to release these bound aromatic compounds. This breaking of the bond is done with the help of enzymes, more precisely enzymes from the group of glycosidases - β-glucosidases. Such release is most often carried out on aromatic varieties in order to express their intensity even more and become more attractive to end consumers. In this work, two enzymes, Lallzyme Beta and Endoyzm β-Split, were investigated on 'Muscat yellow' and 'Muscat rose' wines. After the treatment, the chemical analyzes performed showed a positive influence of the enzymes used on the increase in the concentration of aromatic compounds. A particularly positive impact was shown in compounds such as terpenes and esters, which significantly increased the fruity and floral intensity of the wine. In the end, the sensory evaluation indicated the diversity of the treated wines, and it can be concluded that the use of these enzymes is justified with the aim of producing higher quality wines.

'Modra frankinja' (Vitis vinifera L.) je tradicionalna slovenska sorta grozdja, a kljub temu Slovenci o njej nismo dovolj seznanjeni. Tudi ko gledamo širši vinski svet, opazimo, da o tej sorti in njenih značilnostih vemo zelo malo. Po naših podatkih aromatične tipičnosti vin sorte 'Modra frankinja' še niso bile raziskane. Diplomsko delo je prva tovrstna raziskava te sorte. V delu smo želeli raziskati aromatične lastnosti vin te sorte s pomočjo neformalne senzorične analize. Na ta način smo povzeli katere senzorične note so na splošno izrazite pri obravnavani sorti vina. V nadaljevanju smo s kemijsko analizo aromatičnih spojin vzorcev vina to še kemijsko ovrednotili. Pri kemijski analizi smo se osredotočili na štiri skupine spojin, ki po našem mnenju krojijo tipičnost arom vin sorte 'Modra frankinja'. To so: monoterpeni, estri, norizoprenoidi in C6-alkoholi. Rezultati kažejo, da so prav predstavniki skupine terpenov tisti, ki se pojavljajo v največjih koncentracijah v obravnavanih vinih in pomembno krojijo aromatični profil vin 'Modra frankinja'. C13-norizoprenoidi so bili prisotni v večjih koncentracijah pri vzorcih sortnega vina 'Modra frankinja', kot pri vinih 'Modri pinot' ali vinih bordojskih sort. A še vedno vsebnosti C13-norizoprenoidi ostajajo precej manjše od zaznavnih senzoričnih pragov teh spojin. Although 'Modra Frankinja' (Vitis vinifera L.) is a Slovenian traditional variety, in Slovenia there is poor knowledge available about it. Moreover, also on a global scale very little has been studied and written about this variety or its characteristics. To our best knowledge, the aromatic typicality of the wines made from 'Modra frankinja' grapes has not yet been researched and this work is thus the first to undertake such an investigation. Therefore, we initially set ourselves the goal of exploring general aromatic spectrum by unformal sensory analysis, aiming to get a sense of the notes predominating in the observed wines. This was followed by chemical analysis, to delve into the individual compounds. In the chemical analysis part we focused on four groups of compounds that we thought would play an important role in characterizing the aromas of 'Modra frankinja', namely, monoterpenes, esters, norisoprenoids, and C6-alcohols. The results of our experiment show that the group of terpenes is the one that occurs in the highest concentrations in wines of 'Modra frankinja' and shapes a large part of the aromatic profile. Also, C13-norisoprenoids were higher in the samples of 'Modra frankinja' in comparison with 'Pinot noir' and Bordeaux varieties. However, the levels remain much lower than the reported sensory thresholds.

Ciljevi istraživanja: Definirati optimalne uvjete izolacije kanabinoida i terpena iz cvata industrijske konoplje primjenom “zelene” ekstrakcije ugljikovim dioksidom u superkritičnom stanju (sCO2); kvantificirati dobivene ekstrakte na udio kanabinoida i terpena; te odrediti antioksidacijske i antimikrobne učinke sCO2 ekstrakata. Materijal i metode: Za istraživanje su korišteni osušeni cvatući vršni dijelovi industrijske konoplje (Cannabis sativa L.) sorte Futura 75 s hrvatskog tržišta. Ekstrakcija ciljanih bioaktivnih sastavnica provedena je primjenom sCO2. Eksperimentalna matrica napravljena je prema dizajnu centralnog kompozitnog plana pokusa, a analizom varijance procjenjen je stupanj točnosti primijenjene metodologije. Identifikacija i kvantifikacija isparljivih sastavnica sCO2 ekstrakata provedena je u vezanom sustavu plinske kromatografije i spektrometrije masa (GC-MS), dok su kanabinoidi određeni metodom tekućinske kromatografije visoke djelotvornosti (HPLC-DAD). Antioksidacijska svojstva dobivenih ekstrakata određena su primjenom DPPH testa, dok je antibakterijska aktivnost ispitana mikrodilucijskom metodom za određivanje minimalnih inhibitornih koncentracija. Rezultati: Analiza plinskom kromatografijom i spektrometrijom masa (GC-MS) pokazala je β-mircenski kemotip sCO2 ekstrakata, s visokim udjelom α-pinena i β-pinena te nizom drugih monoterpena s manjim udjelima, dok je najzastupljeniji seskviterpen bio β-kariofilen. Glavni kanabinoidi sCO2 ekstrakata bili su CBDA i CBD, praćeni manje zastupljenim CBCA, CBC, THCA-A, THC, CBGA, CBG, CBDVA, dok CBN i THCVA nisu detektirani. sCO2 ekstrakt dobiven pri tlaku 320 bara i temperaturi 40 °C, s najvećim sadržajem CBDA, imao je najbolja antioksidacijska svojstva. Svi ekstrakti su pokazali dobro antibakterijsko djelovanje na vrste E. coli, P. aeruginosa, B. subtilis i S. aureus. sCO2 ekstrakt s najvećim udjelom CBD-a, koji sadrži i visok udio α-pinena, β-pinena, β-mircena i limonena, pokazao je najbolje antibakterijsko djelovanje. Uspostavljeni su optimalni uvjeti za sCO2 ekstrakciju kanabinoida i terpena iz industrijske konoplje. Temperatura od 60 °C pokazala se optimalnom za sve ispitivane odzive, dok je tlak pokazao različit učinak ovisno o ciljanim spojevima. Za monoterpene optimalan je bio niži tlak, dok je viši tlak bio pogodan za ekstrakciju seskviterpena. Primjena viših tlakova bila je poželjna i za ekstrakciju CBD-a. Postupkom dekarboksilacije kao predtretmana biljnog materijala prije sCO2 ekstrakcije dobiveni su ekstrakti s visokim prinosom CBD-a kao dominantnom sastavnicom. Zaključak: Optimiran je proces sCO2 ekstrakcije za dobivanje ekstrakata industrijske konoplje s antioksidacijskim i antibakterijskim svojstvima te potencijalom topikalne primjene u medicinske i kozmetičke svrhe. Istraživanje je potvrdilo selektivnost primijenjene tehnike za ekstrakciju ciljanih terpena i kanabinoida iz cvatova industrijske konoplje. The objective of the research: To define the optimal conditions for isolation of cannabinoids and terpenes from industrial hemp inflorescences by applying "green" supercritical carbon dioxide extraction (sCO2); to quantify the obtained extracts on the content of cannabinoids and terpenes; and to determine the antioxidant and antimicrobial activities of sCO2 extracts. Material and methods: Dried inflorescences of industrial hemp (Cannabis sativa L.) of Croatia-grown Futura 75 variety were used for the research. Extraction of target bioactive compounds using sCO2 was carried out. The experimental matrix was made according to the central composite design, and the analysis of variance was used to assess the degree of accuracy of the applied methodology. Identification and quantification of volatile compounds of sCO2 extracts was determined by gas chromatography-mass spectrometry combined system (GC-MS), while cannabinoids were determined by high-performance liquid chromatography (HPLC-DAD). The antioxidant properties of the obtained extracts were determined by a DPPH scavenging assay, while antibacterial activity was tested by a microdilution method for the determination of minimal inhibitory concentrations. Results: Analysis with gas chromatography and mass spectrometry (GC-MS) showed β-myrcene chemotype in sCO2 extracts, with a high content of α-pinene and β-pinene and a number of other monoterpenes in smaller proportions, while the most dominant sesquiterpene was β-caryophyllene. The main cannabinoids of sCO2 extracts were CBDA and CBD, followed by less common CBCA, CBC, THCA, THC, CBGA, CBG, and CBDVA, while CBN and THCVA were not detected. sCO2 extract obtained at a pressure of 320 bar and a temperature of 40 °C, with the highest CBDA content, exhibited the best antioxidant properties. All extracts exhibited good antibacterial effect against E. coli, P. aeruginosa, B. subtilis and S. aureus. sCO2 extract with the highest content of CBD, which also contains a high content of α-pinene, β-pinene, β-myrcene and limonene, showed the best antibacterial activity. Optimal conditions have been established for the sCO2 extraction of cannabinoids and terpenes from industrial hemp. The temperature of 60 °C proved to be optimal for all studied responses, while pressure had a different effect depending on the compounds targeted. For monoterpenes, lower pressure was optimal, while higher pressure was suitable for the extraction of sesquiterpenes. The application of higher pressures was also desirable for CBD extraction. By using decarboxylation as a pretreatment of plant material before sCO2 extraction, extracts with a high yield of CBD were obtained. Conclusion: sCO2 extraction process for obtaining industrial hemp extracts with antioxidant and antibacterial activities and with the potential of topical application for medical and cosmetic purposes has been optimized. The provided research confirmed the selectivity of the applied technique for the extraction of targeted terpenes and cannabinoids from industrial hemp inflorescences.

Znanstvene raziskave izvlečkov bele jelke dokazujejo njihov zdravilni potencial na področjih nižanja glukoze v krvi, bolezni srca in ožilja, bolezni sklepov, zdravljenja luskavice in tudi pri zdravljenju raka. Zdravilno moč ji dajejo raznovrstne bioaktivne spojine, kot so npr. polifenoli in med njimi najbolj zastopani lignani. Obstajajo številni postopki pridobivanja izvlečkov bele jelke, med katerimi vse bolj postajajo zanimive tudi zelene tehnologije ekstrakcije, kot je npr. ekstrakcija s subkritično vodo. V magistrski nalogi smo analizirali sestavo izvlečkov bele jelke, pridobljenih z ekstrakcijo s subkritično vodo, in jih primerjali z izvlečki, pridobljenimi z ekstrakcijo s superkritičnim CO2, visokotlačno alkoholno ekstrakcijo z naknadnim spiranjem s superkritičnim CO2, industrijsko vodno ekstrakcijo in laboratorijsko vodno ekstrakcijo. S pomočjo metode tekočinske kromatografije visoke ločljivosti (HPLC) smo analizirali prisotnost lignanov v dobljenih izvlečkih (izolaricirezinola, hidroksimatairezinola, sekoizolaricirezinola, laricirezinola, pinorezinola, matairezinola), s plinsko kromatografijo, sklopljeno z masno spektrometrijo (GC-MS), smo raziskovali prisotnost hlapnih spojin, z metodo Folin-Ciocalteujevega reagenta smo ugotavljali celokupno vsebnost polifenolov in z metodama DPPH in ABTS•+ ugotavljali antioksidativno aktivnost. Ugotovili smo, da je kvalitativna in kvantitativna sestava izvlečkov bele jelke močno odvisna od uporabljene ekstrakcijske metode, prav tako pa tudi od uporabljenega dela drevesa (vej in lubja). Največjo celokupno vsebnost polifenolov, antioksidativno aktivnost in vsebnost ekstrahiranih lignanov smo opazili pri vzorcih, pridobljenih iz lubja bele jelke, najvišje vsebnosti hlapnih spojin (terpenov in terpenoidov) pa pri vzorcih, pridobljenih iz vej bele jelke. Kot najučinkovitejša metoda se je v naši raziskavi izkazala ekstrakcija s subkritično vodo, zato menimo, da ta predstavlja velik potencial na področju pridobivanja izvlečka bele jelke kot tudi drugih rastlinskih izvlečkov. Scientific studies on silver fir extracts prove their healing potential in the fields of lowering blood glucose, cardiovascular diseases, joint diseases, psoriasis treatment and also in the treatment of cancer. Their healing power is given by various bioactive compounds such as polyphenols, which are mainly represented by lignans. There are many extraction procedures for obtaining silver fir extracts among them green extraction technologies have become increasingly popular, such as subcritical water extraction. In the master 's thesis, we analysed the composition of silver fir extracts obtained with subcritical water extraction and compared them with extracts obtained with supercritical CO2 extraction, high-pressure alcohol extraction followed by flushing with supercritical CO2, industrial water extraction and laboratory water extraction. Using high-performance liquid chromatography (HPLC), we analysed lignans in the extracts (isolariciresinol, hydroxymatairesinol, sekoizolariciresinol, lariciresinol, pinoresinol, matairesinol), with gas chromatography-mass spectrometry (GC-MS) we analysed the presence of volatile compounds, with Folin-Ciocalteu method we analysed the total phenolic compounds, and the antioxidative activity of individual extracts was determined by the DPPH and ABTS•+ methods. We found that the qualitative and quantitative composition of silver fir extracts strongly depends on the extraction method, and also on the part of the silver fir tree (branches, bark). The highest total phenolic compounds, antioxidant activity and concentration of extracted lignans were discovered in silver fir bark samples, and the highest content of volatile compounds (terpenes and terpenoids) in silver fir branch samples. The most efficient method in our research was proven to be subcritical water extraction therefore, we conclude that this technique represents great potential in obtaining silver fir extracts and other plant extracts.

Hemp (Cannabis sativa L.) cannabinoids and terpenoids have therapeutic effects on human and animal health. Cannabis plants can often have a relatively high heterogeneity, which leads to different phenotypes that have different chemical profiles despite being from the same variety. Little information exists about cannabinoid and terpenoid profiles in different hemp phenotypes within the same variety. For this study, 11 phenotypes from three different varieties (“Carmagnola” selected (CS), “Tiborszallasi” (TS), and “Finola” selection (FS)) were analyzed. The components of essential oil (29) were analyzed using gas chromatography with flame ionization detection (GC/FID), and 10 different cannabinoids of each phenotype were determined using high-performance liquid chromatography (HPLC). Principal component analysis (PCA) and analysis of variance (ANOVA) showed that according to the components of essential oil, FS and TS plants were more uniform than CS plants, where there were great differences between CI and CII phenotypes. The content of cannabinoid CBD-A was the highest in all four FS phenotypes. By comparing cannabinoid profiles, FS was clearly separated from TS and CS, while these two varieties were not clearly distinguishable. Phenotypes TV and CI had the highest total content of Δ-9-THC, while all phenotypes of FS had the highest total content of CBD. The highest total content of CBG was determined in phenotype CI. Obtained results are useful for the development of new supplementary ingredients, for different pharmacy treatments, and for further breeding purposes.

Procjena botaničkog sastava pašnjaka na različitim područjima s posebnim osvrtom na udjele terpena, koji osim učinka na terpenski profil arome mesa preživača mogu imati i potencijalni imunomodulacijski učinak, od bitnog je interesa suvremenog ovčarstva u R. Hrvatskoj. U istraživanjima u okviru ovog rada, pokazano je da ljekovite svojte bogate terpenima poput gospine trave (Hypericum perforatum L.), stolisnika (Achillea millefolium L.) i matičnjaka (Melissa officinalis L.) zbog statistički značajno veće učestalosti, gustoće i pokrovnosti na pašnjacima Velike Crkvine, u odnosu na pašnjake u Kvarte, potencijalno utječu na bolji imunosni status ovaca. Stoga su u ovom radu dobiveni rezultati znanstvena preporuka za daljnju procjenu biljnog sastava pašnjaka u Velikoj Crkvini tijekom istraživanja HRZZ projekta IP-2016-06-3685 u cilju postizanja dodatnih saznanja o učinku terpena na zdravlje i aroma sastav janjećeg mesa i to sve u svrhu proizvodnje inovativnih funkcionalnih proizvoda od janjećeg mesa., The botanical composition of pastures in various areas, with special regard to terpene proportions, have an effect on the terpene profile of ruminant meat aroma, and may also have a potential immunomodulation effect in the animals. Therefore, assessments of pasture composition are of great interest to modern sheep breeding in Croatia. They study showed that medicinal plants with high terpene content, such as St. John’s Wort (Hypericum perforatum L.), yarrow (Achillea millefolium L.) and lemon balm (Melissa officinalis L.) have a potential effect on improving immune status in sheep. The pastures of Velika Crkva had a statistically significant higher frequency, density and cover of these species than the pastures in Kvarte. Therefore, the results presented here contributed to the scientific recommendation for further assessment of the plant composition of pastures in Velika Crkvina during a separate study the research of the Croatian Science Foundation project IP-2016-06-3685 in order to gain additional knowledge about the effect of terpenes on the health and flavour composition of lamb meat and to contribute to the production of innovative and functional lamb meat products.

Dehydration or drying of grapes is one of the most important steps in the production of Croatian traditional dessert wine Pro&scaron, ek. The natural sun drying of grapes is the traditionally used method in Pro&scaron, ek production. Alternative methods, such as dehydration under controlled conditions, have been studied as safer and faster methods than the traditional sun drying but without precise knowledge of the effect on volatile compounds. The objective of this work was to study how dehydration of grapes carried out in a greenhouse and an environmentally controlled chamber impacts on the free and glycosidically bound volatile compounds of native grape cv. &lsquo, Mara&scaron, tina&rsquo, The 36 volatile compounds were identified and quantified using headspace solid-phase micro extraction coupled with gas chromatography-mass spectrophotometry (HS-SPME-GC/MS). The results showed that the aroma profile of dehydrated grapes was significantly different from that of fresh grapes. Regarding free forms, significant increases in the concentration of 2-methyl-1-propanol, 1-butanol, 2-hexen-1-ol, 1-hexanol, ethyl hexanoate, hexyl acetate, o-cymene, linalool oxide, and terpinen-4-ol and geraniol were found in greenhouse-dried grapes, whereas increases in cis-limonene-epoxide, trans-limonene epoxide, and &gamma, hexalactone were higher in chamber-dried grapes compared to greenhouse-dried grapes. Glycosidically bound forms of o-cymene, linalool oxide, linalool, and terpinen-4-ol were increased in both types of drying, whereas &beta, damascenone was increased only in greenhouse-dried grapes.

Muškatna kadulja (Salvia sclarea L.) je biljka iz porodice usnača (Lamiaceae) koja prirodno raste oko Mediteranskog mora, na području južne Europe i sjeverne Afrike, i u srednjoj Aziji. Cilj ovog rada bio je odrediti profil hlapljivih spojeva muškatne kadulje, i to odvojeno cvjetova, a odvojeno cijele biljke, odnosno odrediti sastav i sadržaj ovih spojeva. Radi što potpunijeg uvida u sastav i sadržaj hlapljivih spojeva za njihovu izolaciju su korištene dvije metode: hidrodestilacija i ekstrakcija vršnih para na čvrstoj fazi. Analiza uzoraka provedena je vezanim sustavom plinska kromatografija-spektrometrija masa (GC/MS). Terpenski spojevi su kvalitativno i kvantitativno najvažniji sastojci eteričnih ulja i vršnih para izoliranih iz cvjetova i biljke muškatne kadulje. Glavni spojevi u oba eterična ulja, izolirana iz biljke i iz cvjetova, su oksidirani monoterpeni linalil-acetat (27,4-30,6%) i linalol (18,2-19,1%). Glavni spojevi vršnih para su linalil-acetat (54,9%) i seskviterpen germakren D (19,7%) u uzorku izoliranom iz cijele biljke, odnosno linalil-acetat (76,6%) i linalol (6,1%) u uzorku izoliranom iz cvjetova. Clary sage (Salvia sclarea L.) is a plant from Lamiaceae family that naturally grows around the Mediterranean Sea, in southern Europe and northern Africa and in central Asia. The aim of this study was to determine the chemical profile of clary sage volatile compounds, from flowers and whole plant separately, and determine the composition and content of these compounds. For more complete insight into the composition and content of volatile compounds two isolation methods were used: hydrodistillation and headspace solid-phase microextraction (HS-SPME). Analysis of volatile samples was carried out by gas chromatography-mass spectrometry (GC/MS). Terpenes are qualitatively and quantitatively the most important ingredients of essential oils and headspaces obtained from clary sage flowers and whole plant. The major constituents of both essential oils, obtained from whole plant and from flowers, were oxygenated monoterpenes linalyl acetate (27,4-30,6%) and linalool (18,2-19,1%). The major constituents of headspaces were linalyl acetate (54,9%) and sesquiterpene germacrene D (19,7%) in sample obtained from whole plant, and linalyl acetate (76,6%) and linalool (6,1%) in sample obtained from flowers.

Vrsta Candida albicans jedan je od najčešćih uzročnika gljivičnih infekcija ljudi. U okviru ove doktorske disertacije ispitivani su mehanizmi antifungalnog delovanja 9 flavonoida (apigenin, apigetrin, astragalin, kvercetin, kvercitrin, izokvercitrin, rutin, luteolin i viteksin), 2 terpena (kamfor i eukaliptol) i 4 nitratna estra heterocikličnih jedinjenja (N-(2-nitrooksietil)-1Η-indol-2-karboksamid (MK55), Ν-(2-nitroksietil)-hinolin-2-karboksamid (MK56), Ν-(2-nitroksietil)-6-(piperidin-1-il)-9H-purin acetamid (MK94), 5΄-Ο-nitro adenozin (MK129)). Najbolju antifungalnu aktivnost ispoljili su nitratni estri heterocikličnih jedinjenja. U smanjenju razvoja virulentnosti ove gljive, nastanku biofilma i formiranju hifalnih ćelija, istakla se aktivnost flavonoida, posebno apigetrina, apigenina i izokvercitrina. Razvoj antifungalne rezistencije C. albicans povezan je sa povećanjem aktivnosti efluks pumpi kodiranih genima MDR1, CDR1 i CDR2. Promene u ekspresiji ovih gena zabeležene su nakon tretmana astragalinom i apigeninom (smanjenje ekspresije CDR1), kamforom (povećanje ekspresije CDR1, smanjenje ekspresije CDR2) i eukaliptolom (povećanje ekspresije CDR1 i CDR2), ukazujući na potencijalna korisna svojstva flavonoida, ali i moguće neželjene posledice primene testiranih terpena. Rezistencija je povezana i sa povećanom ekspresijom ERG11, gena koji kodira protein lanosterol 14α-demetilazu (CYP51), ciljno mesto azolnih lekova. Izokvercitrin i MK94 izazivaju neželjeno povećanje, dok MK55 redukuje nivo ekspresije ovog gena. Mehanizam antifungalnog delovanja MK55 obuhvata i inhibiciju fungalnog proteina CYP51. Narušavanje integriteta ćelijske membrane, kao potencijalnog mehanizma delovanja, potvrđeno je za apigenin, apigetrin i astragalin. Upotreba MK55, MK94, apigenina, apigetrina, rutina, kvercetina, kvercitrina, izokvercitrina, luteolina i kamfora može se smatrati bezbednom, dok se eukaliptol zbog svog citotoksičnog efekta prema ćelijama jetre ne preporučuje za potencijalnu primenu u antifungalnoj terapiji. Candida albicans is among the most common human fungal pathogens. Mode of antifungal action has been tested for 9 flavonoids (apigenin, apigetrin, astragalin, quercetin, quercitrin, isoquercitrin, rutin, luteolin and vitexin), 2 terpens (camphor and eucalyptol) and 4 nitrate esters of heterocyclic compounds (N-(2-nitooxyethyl)-1Η-indol-2-carboxamide (MK55), Ν-(2-nitrooxyethyl)-quinoline-2-carboxamide (MK56), Ν-(2-nitrooxyethyl)-6-(piperidin-1-yl)-9H-purine acetamide (MK94), 5΄-Ο-nitroadenosine (MK129)). The most prominent anticandidal activity has been proven for nitrate esters of heterocyclic compounds. Significant activity of flavonoids, especially apigetrin, apigenin and isoquercitrin, could be noticed in reducing fungal virulence, biofilm formation and hyphal growth. Resistance to antifungal therapy is linked to increased activity of C. albicans efflux pumps encoded by MDR1, CDR1 and CDR2. Disturbance in expression levels of these genes has been observed after treatment with astragalin and apigenin (decreased CDR1 expression), camphor (increased CDR1 and decreased CDR2 expression) and eucalyptol (increased CDR1 and CDR2 expression levels), suggesting possible beneficial antimicrobial effects of flavonoids as well as potential harmful consequences of selected terpenes application. Antifungal resistance is also linked to increased expression of ERG11, gene encoding for lanosterol 14α-demethylase (CYP51), target protein for azole antifungals. Treatment with isoquercitrin and MK94 led to increased expression of this gene, while treatment with MK55 reduced it. Mode of action for MK55 involves inhibition of CYP51 enzyme. Damage of fungal cell membrane as a potential mechanism of antifungal activity has been observed for apigenin, apigetrin and astragalin. Application of MK55, MK94, apigenin, apigetrin, rutin, quercetin, quercitrin, isoquercitrin, luteolin and camphor could be considered safe, based on the results of their cytotoxicity towards the liver cell line, while eucalyptol should be used with serious precaution.

Pelargonije so v Sloveniji ene izmed najbolj priljubljenih okrasnih rastlin, ki za svojo rast potrebujejo sončno do polsončno lego. Tekom zgodovine so gojitelji s pomočjo križanja dosegli veliko pestrost sort, zato jih posledično lahko kupimo v različnih barvah. V grobem jih delimo na pet večjih skupin. Poleg okrasne vrednosti pa pelargonije vsebujejo tudi mnogo sekundarnih metabolitov, ki skrbijo za obarvanost cvetov in značilen vonj pelargonij. Sekundarne metabolite delimo na terpene, fenole in dušik vsebujoče spojine. Znotraj pelargonij so do sedaj odkrili terpene in fenole, o dušik vsebujočih spojin pa ne poročajo. Med terpeni sta najbolj zastopana geraniol in citronelol. Od fenolov so v pelargonijah najbolj zastopani flavonoidi, od njih v večjih količinah izokvercetin, rutin, hiperozid in klorogenska kislina. Eterična olja in hidrolate, pridobljene iz pelargonij, se uporablja v kozmetični, agronomski, prehrambeni in farmacevtski industriji. Dokazano delujejo antifungicidno, antibakterijsko, antikancerogeno in antioksidativno. Pozitivne učinke izvlečkov pelargonije so poznali že v zgodovini, zato so jih uporabljali za zdravljenje diareje, ledvičnih bolezni in za znižanje visoke telesne temperature. Pelargoniums are one of the most popular ornamental plants in Slovenia. For their growth they need sunny to half sunny position. Throughout the history growers have produced a great number of varieties with selection and crossing of Pelargonium plants, that is why they can be bought in many colours. We roughly divide them into five major groups. In addition to the decorative value, pelargoniums also contain many secondary metabolites which cause coloring of flowers and gives them characteristic odor of pelargonium. Secondary metabolites are divided into terpenes, phenolics and nitrogen-containing compounds. Terpenes and phenolics have been discovered so far in genus of Pelargonium, while nitrogen-containing compounds have not been reported. Among terpenes, geraniol and citronellol are most represented. The most numerous phenolics in Pelargonium are flavonoids which belong to the group of polyphenols. Isoquercetin, rutin, hyperoside and chlorogenic acid are the most abundant phenolics. Essential oils and hydrolysates derived from Pelargonium can be used in the cosmetic, agronomic, food and pharmaceutical industries. It has been proven that they posess antifungal, antibacterial, anticarcinogenic and antioxidant properties. The positive effects of pelargonium extracts have already been known in history, and they have been used to treat diarrhea, kidney disease and lowering of high body temperature.

Maslina (Olea europaea L.) je jedna od glavnih poljoprivrednih kultura mediteranskog područja. Otkriveno je da su osim ulja i listovi masline bogat izvor komponenti s farmakološkim učinkom. U ovom radu analiziran je sastav eteričnog ulja i hidrolata u različitim razvojnim fazama masline. Vodenom destilacijom u aparaturi po Clevengeru iz nekoliko uzoraka listova masline izolirana su eterična ulja i hidrolati te identificirani GC/MS analizom. U prvom uzorku eteričnog ulja najzastupljeniji spoj bio je ß-ionon (20.4%), u drugom germakren D (31.2%), u trećem endo-fenhil acetat (12.7%), u četvrtom dokosan (40.1%), u petom n-dodekanol (30.6%) i u šestom kariofilen oksid (17.1%). U uzorcima hidrolata najzastupljeniji spoj u većini uzoraka bio je ß-ionon: u prvom uzorku u količini od 15.9%, u drugom uzorku u količini od 40.3%, u trećem uzorku u količini od 42.8%; u četvrtom uzorku najzastupljeniji spoj bio je biciklogermakren (39.5%), u petom ß-ionon (25.3%) i u šestom najzastupljeniji je bio miristicin (35%). Spojevi koji su izolirani u svim uzorcima listova, ili u eteričnom ulju ili hidrolatu su: α-pinen, ß-kariofilen, α-humulen, allo-aromadendren, ß-ionon, miristicin, palmitinska kiselina, oleinska kiselina te dokosan, trikosan i tetrakosan., Olive (Olea europaea L.) is one of the main agricultural crops of the Mediterranean area. It has been discovered that apart from oil, olive leaves are also a rich source of components with pharmacological effect. In this paper the composition of essential oils and hydrolates was observed in different life stages of olive. By aqueous distillation in Clevenger's apparatus, essential oils and hydrolates were isolated from several olive leaf samples and identified by GC/MS analysis. In the first sample of essential oil the most common compound was β-ionone (20.4%), in second germacrene D (31.2%), in third endo-fenchyl acetate (12.7%), in fourth docosane (40.1%), in fifth n-dodecanol (30.6 %) and in sixth caryophyllene oxide (17.1%). In hydrolates the most common compound in most of the samples was β-ionone: in first sample in the amount of 15.9%, in second sample in the amount of 40.3%, in third sample in the amount of 42.8%; in fourth sample the most common component was bicyclogermacrene (39.5%), in fifth ß-ionone (25.3%) and in sixth myristicin (35%). The compounds that were isolated from all of the samples, either from essential oil or hydrolate, were: α-pinene, ß-caryophyllene, α-humulene, allo-aromadendrene, ß-ionone, myristicin, palmitic acid, oleic acid and docosane, tricosane and tetracosane.

Vrsta Candida albicans jedan je od najčešćih uzročnika gljivičnih infekcija ljudi. U okviru ove doktorske disertacije ispitivani su mehanizmi antifungalnog delovanja 9 flavonoida (apigenin, apigetrin, astragalin, kvercetin, kvercitrin, izokvercitrin, rutin, luteolin i viteksin), 2 terpena (kamfor i eukaliptol) i 4 nitratna estra heterocikličnih jedinjenja (N-(2-nitrooksietil)-1Η-indol-2-karboksamid (MK55), Ν-(2-nitroksietil)-hinolin-2-karboksamid (MK56), Ν-(2-nitroksietil)-6-(piperidin-1-il)-9H-purin acetamid (MK94), 5΄-Ο-nitro adenozin (MK129)). Najbolju antifungalnu aktivnost ispoljili su nitratni estri heterocikličnih jedinjenja. U smanjenju razvoja virulentnosti ove gljive, nastanku biofilma i formiranju hifalnih ćelija, istakla se aktivnost flavonoida, posebno apigetrina, apigenina i izokvercitrina. Razvoj antifungalne rezistencije C. albicans povezan je sa povećanjem aktivnosti efluks pumpi kodiranih genima MDR1, CDR1 i CDR2. Promene u ekspresiji ovih gena zabeležene su nakon tretmana astragalinom i apigeninom (smanjenje ekspresije CDR1), kamforom (povećanje ekspresije CDR1, smanjenje ekspresije CDR2) i eukaliptolom (povećanje ekspresije CDR1 i CDR2), ukazujući na potencijalna korisna svojstva flavonoida, ali i moguće neželjene posledice primene testiranih terpena. Rezistencija je povezana i sa povećanom ekspresijom ERG11, gena koji kodira protein lanosterol 14α-demetilazu (CYP51), ciljno mesto azolnih lekova. Izokvercitrin i MK94 izazivaju neželjeno povećanje, dok MK55 redukuje nivo ekspresije ovog gena. Mehanizam antifungalnog delovanja MK55 obuhvata i inhibiciju fungalnog proteina CYP51. Narušavanje integriteta ćelijske membrane, kao potencijalnog mehanizma delovanja, potvrđeno je za apigenin, apigetrin i astragalin. Upotreba MK55, MK94, apigenina, apigetrina, rutina, kvercetina, kvercitrina, izokvercitrina, luteolina i kamfora može se smatrati bezbednom, dok se eukaliptol zbog svog citotoksičnog efekta prema ćelijama jetre ne preporučuje za potencijalnu primenu u antifungalnoj terapiji. Candida albicans is among the most common human fungal pathogens. Mode of antifungal action has been tested for 9 flavonoids (apigenin, apigetrin, astragalin, quercetin, quercitrin, isoquercitrin, rutin, luteolin and vitexin), 2 terpens (camphor and eucalyptol) and 4 nitrate esters of heterocyclic compounds (N-(2-nitooxyethyl)-1Η-indol-2-carboxamide (MK55), Ν-(2-nitrooxyethyl)-quinoline-2-carboxamide (MK56), Ν-(2-nitrooxyethyl)-6-(piperidin-1-yl)-9H-purine acetamide (MK94), 5΄-Ο-nitroadenosine (MK129)). The most prominent anticandidal activity has been proven for nitrate esters of heterocyclic compounds. Significant activity of flavonoids, especially apigetrin, apigenin and isoquercitrin, could be noticed in reducing fungal virulence, biofilm formation and hyphal growth. Resistance to antifungal therapy is linked to increased activity of C. albicans efflux pumps encoded by MDR1, CDR1 and CDR2. Disturbance in expression levels of these genes has been observed after treatment with astragalin and apigenin (decreased CDR1 expression), camphor (increased CDR1 and decreased CDR2 expression) and eucalyptol (increased CDR1 and CDR2 expression levels), suggesting possible beneficial antimicrobial effects of flavonoids as well as potential harmful consequences of selected terpenes application. Antifungal resistance is also linked to increased expression of ERG11, gene encoding for lanosterol 14α-demethylase (CYP51), target protein for azole antifungals. Treatment with isoquercitrin and MK94 led to increased expression of this gene, while treatment with MK55 reduced it. Mode of action for MK55 involves inhibition of CYP51 enzyme. Damage of fungal cell membrane as a potential mechanism of antifungal activity has been observed for apigenin, apigetrin and astragalin. Application of MK55, MK94, apigenin, apigetrin, rutin, quercetin, quercitrin, isoquercitrin, luteolin and camphor could be considered safe, based on the results of their cytotoxicity towards the liver cell line, while eucalyptol should be used with serious precaution.

Kadulja (Salvia officinalis L.) je samonikla ljekovita i aromatična biljka rasprostranjena u mediteranskog dijelu Hrvatske, a posebno u Dalmaciji. Cilj ovog rada bio je odrediti profil hlapljivih spojeva dalmatinske kadulje, i to odvojeno listova, a odvojeno cvjetova, odnosno odrediti sastav i sadržaj ovih spojeva. Radi što potpunijeg uvida u sastav i sadržaj hlapljivih spojeva pripremljena su tri uzorka hlapljivih spojeva, eterično ulje, vršne pare i apsolut. Analiza uzoraka provedena je plinskom kromatografijom-masenom spektrometrijom (GC/MS). Terpenski spojevi su kvalitativno i kvantitativno najvažniji sastojci eteričnog ulja, vršnih para i apsoluta listova i cvjetova kadulje. Glavni sastojci eteričnog ulja listova kadulje su terpenski alkoholi 1,8-cineol i borneol te keton kamfor, a eteričnog ulja cvjetova alkoholi viridiflorol i izoborneol, ugljikovodik -humulen te keton kamfor. U vršnim parama listova i cvjetova kadulje identificirani su svi spojevi. Svi sastojci vršnih para listova i cvjetova kadulje su terpeni, sa -humulenom kao glavnim sastojkom u vršnim parama listova i -pinenom u vršnim parama cvjetova. Za razliku od eteričnih ulja i vršnih para, apsolut listova kadulje karakterizira visoki udio masnih kiselina, 30,4 % u apsolutu cvjetova i 21,1 % u apsolutu listova. Ipak, kvalitativno i kvantitativno, i u apsolutima kadulje dominiraju terpenski spojevi kao što su epimanool, borneol i - gurjunen u apsolutu listova te epimanool, feruginol, pisiferol, -humulen i -gurjunen u apsolutu cvjetova kadulje. Sage (Salvia officinalis L.) is a wild-growing medicinal and aromatic plant which is widespread in the Mediterranean part of Croatia, especially in Dalmatia. The aim of this study was to determine the chemical profile of Dalmatian sage volatile compounds, separately of leaves and flowers, and determine the composition and content of these compounds. For more complete insight into the composition and content of volatile compounds, three samples of volatile compounds were prepared, namely essential oil, headspace and absolute. Analysis of all samples was carried out by gas chromatography-mass spectrometry (GC/MS). Terpenes are qualitatively and quantitatively the most important ingredients of essential oils, headspaces and absolutes of sage leaves and flowers. The major constituents of leaves essential oil were terpene alcohols 1,8-cineol and borneol as well as ketone camphor, while the main constituents of flowers essential oil were alcohols viridiflorol and isoberneol, hydrocarbon -humulene as well as ketone camphor. All the compounds of leaves and flowers headspace were identified. All constituents of leaves and flowers headspace are terpenes, with -humulene as the main constituent of the leaves headspace and -pinene as the main constituent of the flowers headspace. Opposite to the essential oils and headspaces, sage absolutes were characterized by a high content of fatty acids, 30,4 % in flowers absolute and 21,1 % leaves absolute. However, qualitatively and quantitatively, terpenes were dominating compounds in sage absolutes, such as epimanool, borneol i -gurjunene in leaves absolute and epimanool, ferruginol, pisiferol, -humulene i -gurjunene in sage flowers absolute.

Dobra topljivost eteričnih ulja u drugim biljnim uljima i laka apsorpcija kroz kožu omogućuje im primjenu u medicinskoj masaži i aromaterapiji. Komponente eteričnih ulja male su molekulske mase s 10-15 ugljikovih atoma u molekuli. Najvećim dijelom se sastoje od terpena (monoterpeni i seskviterpeni) i fenilpropanskih derivata. Glavnu grupu terpena u eteričnom ulju jele, bora i kleke čine monoterpeni, kod eteričnog ulja lovora to su oksigenirani monoterpeni, dok se eterično ulje smilja najvećim dijelom sastoji od monoterpenskih i seskviterpenskih ugljikovodika. Eterična ulja bora, jele, kleke, lovora i smilja osvjetljavana su na 420 nm i 366 nm 48 sati u fotokemijskom reaktoru. Osvjetljavanjem se ispitala fotostabilnost komponenata eteričnih ulja uz prisutnost zraka, a sve se pratilo i analiziralo vezanim sustavom plinska kromatografija-masena spektrometrija (GC-MS). Analiza je pokazala da se nakon osvjetljavanja događaju promjene u sastavu eteričnih ulja, degradacija komponenata i nastajanje novih. Osvjetljavana ulja su se usporedila sa standardom, odnosno uzorkom ulja koji nije osvjetljavan kako bi se mogle uočiti razlike u sastavu. Najveće promjene u sastavu uočene su kod eteričnog ulja smilja iz čega proizlazi da je u usporedbi s ostalim uljima, smilje najmanje fotostabilno. To znači da se na eterično ulje smilja najviše treba paziti prilikom upotrebe i skladištenja, odnosno treba ga se čuvati u tamnim bocama s prikladnim čepom, te u tamnim i hladnijim prostorijama kao što su podrumi. Od komponenata koje se mijenjaju u sastavu, najzanimljiviji je γ-kurkumen koji je uz neril-acetat jedan od najdragocjenijih komponenata eteričnog ulja smilja. Iz kromatograma frakcije obogaćene γ-kurkumenom nakon osvjetljavanja u trajanju od 48 sati vidi se smanjenje γ-kurkumena, odnosno povećanje drugih komponenata koje su njegovi mogući fotoprodukti. Do sada je jako malo provedenih istraživanja na ovu temu opisanih u literaturi i općenito ima malo informacija o fotostabilnosti eteričnih ulja i sličnih spojeva. Good solubility of essential oils in other vegetable oils and easy absorption through the skin allows them to be used in medical massage and aromatherapy. Essential oils are small molecular masses with 10-15 carbon atoms in the molecule. They mostly consist of terpenes (monoterpenes and sesquiterpenes) and phenylpropane derivatives. The main group of terpenes in essential oil of fir, pine and juniper are monoterpenes, while in the essential oil of laurel main group of terpens are oxygenated monoterpenes. Essential oil of immortelle is mostly composed of monoterpene and sesquiterpene hydrocarbons. Essential oil of pine, fir, juniper, laurel and immortelle were illuminated at 420 nm and 366 nm in chemical reactor. Photostability of essential oils was determined by 48 hour long illumination in the presence of air. Process was monitored and analyzed by the linked gas chromatographic-mass spectrometry (GC-MS) system. The analysis has shown that illumination affects on the changes in the composition of essential oils, component degradation and the formation of new ones. Illuminated oils were compared to the standard, which is an oil sample that is not illuminated, in order to notice the differences in the composition. The most significant changes in composition were seen in the essential oil of immortelle, which means that essential oil of immortelle compared to other oils is the least photostable. Poor photostability means that with the essential oil of immortelle should be carefully handeld. It should be kept in dark bottles with a suitable stopper and stored in dark and cold rooms such as basements. Among the components that change, the most interesting is γ-curcumen which with neril acetate is one of the most valuable components of the essential oil of immortelle. From chromatogram of fraction which is enriched by γ-curcumene after 48 h long illumination, reduction of γ-curcumene can be spotted. Also, number of other components that are possible product of γ-curcumene decomposition increases. There is few researches on this subject in the literature and generally there is a small amount of information on the photostability of essential oils and similar compounds.

Rad ne sadrži sažetak.

Terpeni su najviše zastupljeni kemijski spojevi u industriji parfema i najveći dio mirisnih spojeva čine upravo terpeni. Ovaj rad obrađuje terpene i spojeve koji su dobiveni iz njih a koji se koriste u parfemima ili proizvodima za svakodnevnu upotrebu u koje se dodaju parfemi kako bi se poboljšao njihov miris. U uvodu je opisana uloga terpena u biljkama, fizikalna i kemijska svojstva terpena te njihova upotreba u proizvodima koje svakodnevno koristimo, od kojih su najvažniji parfemi i proizvodi koji ih sadrže. Terpeni se pojavljuju u prirodi te je objašnjena njihova biosinteza i metode izolacije iz prirodnih izvora. Kako je izolacija prirodnih parfemskih sastojaka često komplicirana i ekonomski neisplativa, pojavila se potreba za njihovom sintezom. Zahvaljujući uspješnoj sintezi terpena i drugih mirisnih kemijskih spojeva, parfemi su postali široko dostupni. U radu je poseban naglasak na terpenima, kao i kemijskim spojevima dobivenima iz njih, koji su važni za industriju parfema. Terpeni su podijeljeni u osnovne skupine, navedeni su najvažniji predstavnici svake skupine i reakcije pomoću kojih se dobivaju. Na kraju rada je osvrt o utjecaju parfemskih sastojaka, uključujući i terpene, na ljudsko zdravlje.

Terpeni su velika grupa prirodnih spojeva opće formule (C5H8)n. Kroz povijest su se upotrebljavali i još se upotrebljavaju u razne svrhe i u svim područjima života kao što su: proizvodnja parfema, medicina, prehrambena industrija. Poznato je oko 30 000 različitih terpena (Baran, Maimone, 2007.), a njihova osnovna struktura izgrađena je od 2- metilbutanske jedinice koja se često naziva i izoprenska jedinica. Na temelju broja n u općoj formuli (C5H8)n ili broja C atoma terpeni se dijele na hemiterpene (C5), monoterpene (C10), sesquiterpene (C15), diterpene (C20), sesterpeni (C25), triterpeni (C30), tetraterpeni (C40), politerpeni (>C 40). Terpeni se često nazivaju i izoprenoidi. S obzirom na funkcijske skupine terpene uglavnom čine ugljikovodici, alkoholi i njihovi glikozidi, eteri, aldehidi, ketoni, karboksilne kiseline i esteri. Terpens are a large group of natural organic compounds of general formula (C5H8)n. Throughout history they have been used and theystill are used for various purposes and in all areas of life such as: the production of parfume, medicine, food industry. It is known about 30,000 different terpenes ( Baran, Maimone, 2007.), and their basic structure is built of 2-methylbutanoic unit is often called isoprene units. They can be classified on the basis of value of n or number of carbon atoms present in the structure into hemiterpene (C5), monoterpenes (C10), sesquiterpene (C15), diterpenes (C20), sesterpens (C25), triterpenes (C30), tetraterpens (C40), polyterpenes (>40C). Terpenes are often called isoprenoids. Given the nature of the functional group terpens mainly consist: hydrocarbons, alcohols and their glycosides, ethers, aldehydes, ketons, carboxylic acids and esters.

In order to determine the terpenes effect on enzyme activity is been studied the impact of aqueous (prepared manually (I1) or ultrasound (I2) mixing) and DMSO (dimethyl sulfoxide) terpenes solution (carvacrol, linalool, nerol and geraniol) on the activity of creatine kinase, lactate dehydrogenase, alkaline phosphatase, gamma-glutamyl transferase, aspartate aminotransferase and alanine aminotransferase,as well as the impact of the same type of terpenes solution (carvacrol, linalool, nerol, geraniol, β-pinene, myrcene, citral, eucalyptol, terpineol, and eugenol) on the activity of amylase in the control sera (with the normal (N) and increased (H) activity of the ingredients. Measurement of activity was conducted after incubation of the serum sample from the 10 minutes and aqueous solutions of terpenes prepared as I1 or as I2, or solutions of terpenes in DMSO at four concentrations. All the terpenes solutions have demonstrated that the inhibitory effect depends on various parameters: kinds of serum (higher percentage of inhibition in N than the H serum), the type of solvent ( the aqueous solution as I2 and DMSO solutions showed approximately the same percentage of inhibition in both types of serum) and way of preparing the aqueous solutions (a higher percentage of inhibition have the aqueous solution of terpenes as I2 in N serum and as I1 in H serum), as well as on the type of terpenes or enzymes. Statistical data processing all terpenes solution (with the partial exception of neroli and geraniol) confirmed the grouping or by type of serum or by type of solvent or the method of preparation of aqueous solution.

Electroantennographic (EAG) responses from adults of the chestnut gall wasp, Dryocosmus kuriphilus were recorded on stimulation with a given dose of 22 volatile compounds previously identified from Castanea sativa leaves. All compounds elicited measurable EAGs. The largest EAG amplitudes were recorded in response to the oxygenated monoterpenes, α-terpineol and linalool, and the green leaf volatiles (GLVs), (E)-2-hexenal, (Z)-3-hexen-1-ol, (E)-3-hexen-1-ol and (E)-2-hexen-1-ol. EAGs to hydrocarbon monoterpenes and aromatics were generally weak. In preliminary Y-tube olfactometer bioassays the GLVs, hexyl acetate, (E)-2-hexen-1-ol and (E)-3-hexen-1-ol were attractive when individually tested compared to a solvent control suggesting their possible role in the D. kuriphilus host plant selection. Identification of host plant attractants is useful to develop semiochemically based pest control strategies of the chestnut gall wasp. his research was partly supported by the Campania Region, Contribution number 28/08 to the research program: “Studio per il controllo ecocompatibile del Cinipide del castagno” and by the Lazio Region, research program “Convenzione triennale tra la Regione Lazio e il Dipartimento di Protezione delle Piante dell’Università della Tuscia di Viterbo per determinazioni, studi, sperimentazioni e consulenze entomologiche specialistiche”

Isparljivi sastojci janjećeg mesa iz različitih zemljopisnih područja korišteni su u ovom istraživanju kao eventualni biomarkeri, odnosno poveznica između janjetine i florističkog pokrova određenog uzgojnog područja. U tu svrhu izvršene su analize isparljivih spojeva pečene janjetine porijeklom iz pet različitih područja (dalmatinsko zaleđe, otok Pag, Lika, otok Cres i Kupreško polje u BiH) na GC-MS (SPME metoda). Sva je janjad bila porijeklom od izvornih pasmina ovaca (dalmatinske pramenke, paške ovce, creske ovce, ličke pramenke i kupreške pramenke) iz prethodno navedenih uzgojnih područja. Analizom je utvrđeno 121 različitih isparljivih sastojaka u svih pet vrsta janjetine, od čega 88 u dalmatinske janjetine (DL), 52 u paške janjetina (PL), 70 u ličke janjetine (LL), 76 u creske janjetine (CL) i 66 u kupreške janjetine (KL). Najzastupljenija skupina isparljivih sastojaka bili su aldehidi (oko 50% ukupne površine vršnih područja, ovisno o vrsti janjetine), alkoholi (od 20 do 30% za sve skupine, osim KL koja je sadržavala u prosjeku 7, 7%) i ketoni (oko 10% za sve skupine, osim CL koja je sadržavala u prosjeku 5, 84%). Rezultati ovog istraživanja su uglavnom u skladu s relevantnim referencama u kojima se 2, 3-oktanedion, 3-metilindol (skatol), terpeni kao i dugolančani alkani i C- 7 aldehidi navode kao pouzdani markeri pašnjačkog uzgoja ovaca.

Kemijska analiza terpenov iglic in njihova zastopanost v vretenih Picea abies (L.) Karst

U ovom radu analizirana je fitohemijska i molekularna varijabilnost i diferencijacija prirodnih populacija taksona sekcije Juniperus roda Juniperus (Cupressaceae) sa Balkanskog poluostrva... This thesis deals with phytochemical & molecular variability variability & differentation of natural population of taxa belonging to section Juniperus of the genus Juniperus (Cupressaceae) from the Balkan peninsula...

U ovom radu analizirana je fitohemijska i genetiţka varijabilnost i diferencijacija sedam autohtonih populacija Pinus nigra u Srbiji koje pripadaju razliţitim infraspecijskim taksonima (ssp. nigra, var. gocensis, ssp. pallasiana i var. banatica). U dvogodišnjim ţetinama 195 jedinki iz sedam populacija odreŤen je sadrţaj i varijabilnost n-alkana i terpena. Kutikularne voskove ţetina istraţivanih populacija crnog bora odlikuje serija nalkana od C16 do C33, osim u populacijama opisanim kao ssp. nigra u kojima je detektovana serija od C18 do C33. Najzastupljeniji su n-alkani sa neparnim brojem C atoma: C23, C25, C27 i C29 (proseţni sadrţaj 10.3, 16.5, 15.1 i 10.7%, respektivno). U etarskim uljima ţetina utvrŤeno je ukupno 75 komponenti, od koji je 58 identifikovano. Dominantne komponente su α-pinen (43.6%) i germakren D (29.8%), koje zajedno ţine 73.4% mase ulja. Multivariacione statistiţke analize (PCA, CDA i HCA) koriššenih fitohemijskih markera su, generalno, pokazale tendenciju formiranja tri grupe populacija (nigra, pallasiana i banatica). Na osnovu varijabilnosti terpena, populacije Priboj i Goţ (determinisane kao var. gocensis) pokazale su umerenu tendenciju odvajanja od prve grupe (ssp. nigra), dok je prema sastavu n-alkana populacija Priboj pokazala još vešu tendenciju odvajanja. Pretpostavljeno je da takson gocensis predstavlja tranzicionu formu izmeŤu taksona nigra na zapadu i pallasiana na jugoistoku Srbije. U cilju odreŤivanja genetiţkog diverziteta i diferencijacije sedam istraţivanih populacija P. nigra iz Srbije, testirano je 114 jedinki pomošu jedarnih (nuDNK), hloroplastnih (cpDNK) i mitohondrijalnih (mtDNK) markera koji imaju bi-parentalno, paternalno i maternalno nasleŤivanje, respektivno, kao i razliţitu brzinu evolucije. Zbog nedostatka jedarnih mikrosatelita (nuSSRs) specijalno razvijenih za P. nigra, uraŤena je inter-specijska PCR amplifikacija tri EST-SSRs razvijenih za P. taeda. Iako su dva ESTSSR lokusa uspešno PCR-amplifikovana u našim uzorcima, utvrŤeno je odsustvo varijabilnosti zbog ţega su iskljuţeni iz dalje analize. Od pet testiranih cpDNK SSRs, tri su uspešno PCR-amplifikovana u našim uzorcima, sa 22 detektovana alela koji formiraju 38 haplotipova. Kao što je i oţekivano, s obzirom na intenzivan protok gena posredstvom polena kod ţetinara, utvrŤen je veoma visok nivo cpDNK diverziteta (HT = 0.949) i mali stepen diferencijacije (Fst = 0.017, p = 0.108, Gst = 0.024, Rst = 0.066)... In this work phytochemical and genetic variability and differentiation of seven native populations belonging to different infraspecific taxa of Pinus nigra (ssp. nigra, var. gocensis, ssp. pallasiana i var. banatica) in Serbia were analyzed. In the needles of 195 trees from seven populations, composition and variability of n-alkanes and terpenes were determined. The size of the n-alkanes found in the needle cuticular waxes of the black pine populations investigated ranged from C16 to C33, with the exception of that of the subspecies nigra, which ranged from C18 to C33. The most abundant n-alkanes were the four odd-numbered n-alkanes C23, C25, C27, and C29 (mean contents of 10.3, 16.5, 15.1, and 10.7%, respectively). In the needle essential oils a total of 58 individual components out of the 75 compounds detected could be identified. The major components of the oils were α-pinene (43.6%) i germakrene D (29.8%), comprising together in average 73.4% of the essential oil composition. Multivariate statistical analysis (PCA, CDA, and HCA) of used phytochemical markers showed, in general, a tendency to the formation of three populations groups (nigra, pallasiana, and banatica). Based on the terpene variability, populations Priboj and Goţ, determined as var. gocensis, showed a moderate tendency of separation from the first group (ssp. nigra), while based on n-alkane composition, population Priboj showed even a greater tendency of separation. It was assumed that taxon gocensis might be a transitional form between taxa nigra occurring in the west and pallasiana growing in the southeast of Serbia. We screened 114 trees from seven studied P. nigra populations from Serbia with nuclear (nrDNA), chloroplast (cpDNA) and mitochondrial (mtDNA) markers which have bi-parental, paternal and maternal inheritance, respectively, and different evolutionary rates to infer levels of genetic diversity and structuring within studied area. Due to the lack of nuclear microsatellites (nuSSRs) specifically developed for P. nigra, we employed three EST-SSRs developed for P. taeda for cross-species amplification in P. nigra. Although two EST-SSR loci were successfuly PCR-amplified in our samples, they were invariable and discarded from further analyses...

In studies of population variability, particular attention has to be paid to the selection of a representative sample. The aim of this study was to assess the size of the new representative sample on the basis of the variability of chemical content of the initial sample on the example of a whitebark pine population. Statistical analysis included the content of 19 characteristics (terpene hydrocarbons and their derivates) of the initial sample of 10 elements (trees). It was determined that the new sample should contain 20 trees so that the mean value calculated from it represents a basic set with a probability higher than 95 %. Determination of the lower limit of the representative sample size that guarantees a satisfactory reliability of generalization proved to be very important in order to achieve cost efficiency of the research. Cilj ovih istraživanja je da se na osnovu varijabilnosti hemijskih svojstava početnog uzorka proceni veličina novog reprezentativnog uzorka jedne populacije munike. Statističkom analizom je obuhvaćen sadržaj 19 svojstava (terpenskih ugljovodonika i njihovih derivata) početnog uzorka od 10 elemenata (stabala). Utvrđeno je da novi uzorak treba da sadrži 20 stabala da bi srednja vrednost izračunata iz njega reprezentovala osnovni skup sa verovatnoćom većom od 95 %. Određivanje donje granice veličine reprezentativnog uzorka koja garantuje zadovoljavajuću pouzdanost uopštavanja je veoma važno zbog ekonomičnosti istraživanja. Projekat ministarstva br. OI-173011, br. TR-37002 i br. III-43007

Biljni sekundarni metaboliti su vrlo raznolike vrste molekula prisutne u većini vaskularnih biljaka. Ti prirodni biljni spojevi ne sudjeluju u primarnom metabolizmu biljke. U većim koncentracijama štite biljku od biljojeda i raznih štetnika, djelujući negativno na njihove stanične i metaboličke procese. U nižim dozama i koncentracijama mogu imati potpuno blagotvoran utjecaj na ishranu i zdravlje životinja. U ovom radu donosimo moguće učinke biljaka s ljekovitim svojstvima na hranidbeno ponašanja životinja koje nastaje kao posljedica ishrane mediteranskm grmovima bogatim sekundarnim biljnim metabolitima. Taj mehanizam – hranidbeno ponašanje životinja uvjetovano posljedicama – ukazuje da životinje ispašom u mediteranskim pašnjačko-šumskim sastojinama s raznolikim biljnim vrstama mogu zadovoljiti svoje prehrambene potrebe samostalnim odabrom biljaka s ljekovitim svojstvima kako bi se neutraliziralo negativno djelovanje biljaka s visokim sadržajem nepoželjnih biljnih spojeva. Razumijevanje ponašanja životinja pri ishrani različitim biljnim vrstama važno u je prvom redu radi zdravstvenog stanja i dobrobiti životinja, smanjenja razine stresa i straha, te mogućnosti otkrivanja ranog stadija bolesti životinja. Stoga, strategija ispaše u biokemijski / biološki raznolikim prirodnim ekosustavima trebala bi omogućiti preživačima da ispolje svoje hranidbene sklonosti prema svim biljnim vrstama prisutnim u pašnjačkoj zajednici., Plant secondary metabolites (also called phytotoxins; natural products) are molecules made in most vascular plants that are not necessary for their survival, but in higher concentrations protect plants from consumers or from pathogens. These phytochemicals can adversely affect cellular and metabolic processes in herbivores, but also at low doses and in appropriate mixtures, they can have beneficial effects on animal nutrition, health and other therapeutic impacts. In this paper, we demonstrate the potential effects of plants with medicinal properties on animal foraging behavior as a function of the consequences they experience after ingestion of Mediterranean shrubs that are rich in phytotoxins. This mechanism – behavior by consequences – suggests animals are able to meet nutritional requirements and self-select certain plants with medicinal properties if they are offered the opportunity to do so when foraging in diverse Mediterranean shrub communities. Understanding the feeding behavior of ruminants when offered a variety of plant species is necessary to be able to improve their health and well-being by reducing levels of stress and fear; it may also lead to the early detection of illness. Thus, management strategies in biochemically/biologically diverse ecosystems should benefit from allowing ruminants to manifest their feeding preferences.

Biljni sekundarni metaboliti su vrlo raznolike vrste molekula prisutne u većini vaskularnih biljaka. Ti prirodni biljni spojevi ne sudjeluju u primarnom metabolizmu biljke. U većim koncentracijama štite biljku od biljojeda i raznih štetnika, djelujući negativno na njihove stanične i metaboličke procese. U nižim dozama i koncentracijama mogu imati potpuno blagotvoran utjecaj na ishranu i zdravlje životinja. U ovom radu donosimo moguće učinke biljaka s ljekovitim svojstvima na hranidbeno ponašanja životinja koje nastaje kao posljedica ishrane mediteranskm grmovima bogatim sekundarnim biljnim metabolitima. Taj mehanizam – hranidbeno ponašanje životinja uvjetovano posljedicama – ukazuje da životinje ispašom u mediteranskim pašnjačko- šumskim sastojinama s raznolikim biljnim vrstama mogu zadovoljiti svoje prehrambene potrebe samostalnim odabrom biljaka s ljekovitim svojstvima kako bi se neutraliziralo negativno djelovanje biljaka s visokim sadržajem nepoželjnih biljnih spojeva. Razumijevanje ponašanja životinja pri ishrani različitim biljnim vrstama važno u je prvom redu radi zdravstvenog stanja i dobrobiti životinja, smanjenja razine stresa i straha, te mogućnosti otkrivanja ranog stadija bolesti životinja. Stoga, strategija ispaše u biokemijski / biološki raznolikim prirodnim ekosustavima trebala bi omogućiti preživačima da ispolje svoje hranidbene sklonosti prema svim biljnim vrstama prisutnim u pašnjačkoj zajednici.

Materiali rastlinskega izvora, med njimi tudi navadna smreka (Picea abies), vsebujejo substance, ki nastajajo v presnovi med razvojem rastline in se kopičijo v različnih delih rastlin. Prav tako te zdravilne rastline vsebujejo spojine, ki imajo številne specifične farmakološke in tehnološke vrednote. V diplomski nalogi smo raziskali različne postopke izolacije hlapnih aromatskih substanc (α- in β- pinena) iz lubja, žagovine in iglic navadne smreke. Izolacija monoterpenov je potekala s postopkom konvencionalne operacije vodne destilacije s Clevenger aparatom ter ekstrakcijskim postopkom visokotlačne ekstrakcije s superkritičnim ogljikovim dioksidom v laboratorijskem in pilotnem merilu. Visokotlačnim ekstrakcijam smo z dinamično metodo, določili optimalne obratovalne pogoje ter vse ekstrakte kvantitativno analizirali s plinsko kromatografijo v povezavi z masno selektivnim detektorjem. Po analizi ekstraktov smo materiale ekstrahirali pri optimalnih obratovalnih pogojih na visokotlačni napravi UHDE v pilotnem merilu. Materials of plants origin including spruce (Picea abies) contain substances that occur during growth of plant and are stored in its various parts. In addition, these medicinal plants contain compounds with many specific pharmacological and technological properties. In thesis we studied various processes of isolation of volatile aromatic substances (α- and β- pinene) from the bark, sawdust and needles of the pine. Isolation of monoterpen has been performed by the process of conventional technical operation with water distillation made by special device named Clevenger and extraction procedure with supercritical carbon dioxide in so - called laboratory and pilot scale. The high - pressure extraction was performed by dynamic method and optimal operation conditions were determined. Optained extracts were analayzed by gas chromatography coverted to mass selective detector. After analysis of extracts were extracted at optimal materials in the best operating conditions by the high pressure device UHDE in so - called pilot scale.

Utilization of the Mediterranean shrubby vegetation is often limited by secondary compounds, such as terpenes, which at too high concentrations can adversely affect forage intake and animal health. Ingesting compounds such as activated charcoal and energy can ameliorate the negative effects of secondary compounds and enable animals to eat more shrubs. Thus, our objectives were to determine if supplemental charcoal, energy and numbers of shrub species offered influenced intake of shrubs by sheep and goats. We conducted three experiments each with 12 lambs and 12 kids (6 activated charcoal vs. 6 controls). In first experiment, we initially offered three shrubs (Juniperus phoenicea, Helichrysum italicum and Juniperus oxicedrus), ) then in second experiment, two shrubs (Juniperus phoenicea and Helichrysum italicum), and finally one shrub (Juniperus phoenicea) in third experiment. In all thee experiments (Exp. 1, P

Ethnic Agave alcoholic beverages such as raicilla, sisal, tequila, mezcal, bacanora, sotol and pulque have been analyzed by gas chromatography and headspace solid-phase microextraction- gas chromatography-mass spectrometry (HS-SPME-GC-MS). There were 105 compounds identified, eleven were classified as major compounds and the others were classified as minor compounds. Seventeen minor compounds could be used as authenticity markers since they were beverage specific. Cluster analysis (CA) showed that Agave alcoholic beverages could be distinguished by multivariate analysis of major compounds; however, the analysis of minor compounds provided a better fingerprinting., Plinskom kromatografijom i mikroekstrakcijom na čvrstoj fazi, u kombinaciji s plinskom kromatografijom i masenom spektrometrijom (HS-SPME-GC-MS), ispitani su uzorci tradicionalnih meksičkih pića proizvedenih iz Agave (raicilla, sisal, tequila, mezcal, bacanora, sotol i pulque). Identificirano je 105 spojeva, od čega 11 glavnih sastojaka. Utvrđeno je da se za markere autentičnosti pojedinih pića može upotrijebiti 17 sporednih sastojaka. Razvrstavanjem uzoraka (engl. cluster analysis – CA) dokazano je da se tradicionalna meksička pića proizvedena iz Agave mogu razlikovati multivarijantnom analizom glavnih sastojaka, a analiza sporednih sastojaka omogućava točnije utvrđivanje porijekla pića.

The effect of red wine phenolic extracts on the stability of wine volatile esters and terpenes was examined. Muscat (white) and Xinomavro (red) wines were enriched with each extract at 120 or 200 mg/L, and stored in open bottles at 20 °C for 3 and 2.5 days, respectively. Moreover, a model wine medium containing isoamyl acetate, ethyl hexanoate and linalool was enriched with each extract at 100 mg/L, and stored in sealed bottles at 20 °C for 45–90 days. All samples were analysed for volatiles using SPME along with GC-MS analysis. Phenolic composition of wine extracts was determined using HPLC-DAD. No effect on the concentration of any volatiles was observed as a result of the addition of each extract in each wine or the model medium. A wine extract rich in phenolic acids and another one rich in anthocyanins and flavanols inhibited the decrease of volatile esters and terpenes in one or both wines and the model medium. Among them were several important for the aroma of wine such as ethyl acetate, isoamyl acetate, ethyl hexanoate, ethyl octanoate, ethyl decanoate and linalool. The results presented here indicate that wine phenolic acids, and anthocyanins or flavanols may be taken into account as potent inhibitors of the disappearance of volatile esters and terpenes in wines., Istražen je utjecaj fenolnih ekstrakata crnog vina na stabilnost hlapljivih estera i terpena u vinu. Muškat (bijelo vino) i Xinomavro (crno vino) obogaćeni su s po 120 ili 200 mg/L ekstrakta i skladišteni u otvorenim bocama tijekom 3 odnosno 2,5 dana pri 20 °C. Pripravak sličan vinu, koji sadrži izoamilacetat, etilheksanoat i linalol, također je obogaćen s 100 mg/L fenolnog ekstrakta, te skladišten u zatvorenim bocama pri 20 °C tijekom 45-90 dana. Hlapljive komponente svih uzoraka analizirane su SPME i GC-MS analizom. Fenolni sastav vinskih ekstrakata određen je HPLC-DAD metodom. Dodatkom fenolnih ekstrakata nije zabilježena promjena koncentracije hlapljivih sastojaka u vinima i pripravku sličnom vinu. Vinski ekstrakt bogat fenolnim kiselinama i drugi bogat antocijanima i flavanolom smanjili su razgradnju hlapljivih estera i terpena u vinu i pripravku, prije svega spojeva važnih za aromu vina, kao što su etilacetat, izoamilacetat, etilheksanoat, etiloktanoat, etildekanoat i linalol. Rezultati pokazuju da bi se fenolne kiseline, antocijanini i flavanoli iz vina mogli upotrijebiti za inhibiciju razgradnje hlapljivih estera i terpena tijekom skladištenja vina.