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Naravne snovi so spojine ali ekstrakti, ki izvirajo iz živih organizmov ter navadno izkazujejo biološko delovanje. So osnovna surovina za zdravila naravnega izvora. V farmakognoziji se ukvarjamo z njihovim izborom, karakterizacijo, identifikacijo in uporabo. Pogost pristop k odkrivanju potencialno zdravilnih naravnih snovi je preizkušanje učinkov rastlinskih izvlečkov, ki nas v povezavi s kromatografskimi separacijami vodi do izolacije biološko aktivnih molekul. Njihove kemijske strukture ugotavljamo s sodobnimi spektroskopskimi in spektrometrijskimi metodami in nadaljujemo s preizkušanjem njihovega farmakološkega delovanja. Ajda predstavlja bogat vir hranil z ugodnim vplivom na zdravje, zato njena poraba v skladu s popularnostjo zdravega načina življenja narašča. Poleg zdravju koristnih snovi pa ajda vsebuje tudi manj znane, fototoksične fagopirine, ki pod vplivom svetlobe oksidirajo biološko snov in povzročijo njene poškodbe. Za zagotavljanje varnosti ajde in njenih izdelkov je treba fagopirine vrednotiti tako s prehranskega kot s farmacevtskega vidika. Ne nazadnje gre tudi za spojine s potencialno terapevtsko uporabnostjo, na primer v fotodinamični terapiji. Razvili smo učinkovit postopek ekstrakcije fagopirinov z uporabo raztopine acetona in vode. Ugotovili smo, da so najprimernejši material za pridobivanje fagopirinov ajdovi cvetovi. Razvili smo preprosto in občutljivo metodo tekočinske kromatografije visoke ločljivosti s fluorescentno detekcijo. Metoda je primerna za analizo vzorcev ajde in kot izhodišče za razvoj preparativne metode za izolacijo fagopirinov. Ugotovili smo, da je poleg osnovne molekule fagopirina F v zeli ajde še vsaj sedem njegovih derivatov. Te derivate smo kromatografsko ločili na posamezne frakcije. Vrednotili smo jih z UV-Vis absorpcijo, masno spektrometrijo in NMR spektroskopijo. Ugotovili smo strukturi dveh novih fagopirinov, ki se razlikujeta po skupinah, pripetih na naftodiantronski obroč. Fagopirin F je že predhodno znana spojina z dvema piperidinskima obročema, fagopirin E je spojina s piperidinskim in pirolidinskim obročem, fagopirin A pa je spojina z dvema pirolidinskima obročema. Dokazali smo, da v ajdi ni fagopirinov, temveč zgolj protofagopirini kot njihove predstopnje z necikliziranim naftodiantronskim obročem. Po raztopitvi v ekstrakcijskem topilu in pod vplivom svetlobe se protofagopirini pretvorijo v fagopirine. Raziskovali smo topnost, fluorescenco in UV-Vis spektre fagopirinov v različnih topilih iz različnih vzorcev ajde. Za kvantitativno vrednotenje vsebnosti fagopirinov je treba pridobiti čiste spojine kot referenčne spojine. Raziskovalci namesto fagopirina kot referenčne spojine uporabljajo hipericin iz šentjanževke, ki pa zaradi nekoliko drugačne strukture in s tem drugačne absorpcije pri spektrofotometrijski detekciji ni ustrezen za točno kvantitativno vrednotenje fagopirinov. Naše ugotovitve in novo razvite metode bodo pripomogle k nadaljnjemu raziskovanju fagopirinov in vrednotenju vsebnosti in varnosti ajdovih izdelkov. V drugem delu doktorskega dela smo se ukvarjali z izvlečki debla navadne jelke. Izvlečki debel različnih vrst borovcev so znani po vsebnosti polifenolnih spojin z zanimivimi farmakološkimi učinki, predvidoma povezanimi z antioksidativnim delovanjem. Najbolj znan, raziskan in uporabljan je izvleček lubja obmorskega bora (Pinus maritima), Pycnogenol®. Ugotovili smo, da so izvlečki navadne jelke učinkovitejši antioksidanti od Pycnogenola® tako v in vitro antioksidativnih testih, kot tudi na humanih celičnih linijah, izoliranih iz periferne krvi. Komponente izvlečka lubja smo kromatografsko ločili z normalnofazno kolonsko kromatografijo pri zvišanem tlaku in reverznofazno tekočinsko kromatografijo visoke ločljivosti. Vsebovane spojine smo identificirali z metodami masne spektrometrije, UV-Vis absorpcijske spektroskopije in s primerjavo z referenčnimi spojinami. Identificirali smo šest fenolkarboksilnih kislin (galno, homovanilno, protokatehujsko, p-hidroksibenzojsko, vanilno in p-kumarno), tri flavonoide (katehin, epikatehin in katehintetrametileter) in štiri lignane (taksiresinol, 7-(2-metil-3,4-dihidroksitetrahidropiran-5-iloksi)-taksiresinol, sekolariciresinol in lariciresinol). Nadaljevali smo s farmakološkim testiranjem. Ugotovili smo, da izvleček zmanjša nastajanje ateroskleroze pri budrah na kronični aterogeni dieti. Ovrednotili smo vpliv izvlečka in dveh v njem prisotnih fenolnih kislin, p-kumarne in protokatehujske, na izolirana podganja srca, podvržena akutnim ishemijsko-reperfuzijskim poškodbam. Ugotovili smo, da je izvleček značilno zmanjšal poškodbo srčnega tkiva, merjeno preko sproščanja laktat-dehidrogenaze in zmanjšal pojavnost aritmij. p-kumarna in protokatehujska kislina nista imeli tako izrazitih učinkov, zato verjetno samo delno prispevata k sinergističnemu delovanju v izvlečku prisotnih učinkovin. Kot ustrezno surovino za raziskovanje in uporabo smo oblikovali izvleček, raztopljen v polietilenglikolu. Pojavila se je potreba po ugotavljanju vsebnosti vode v tovrstnih vzorcih. Uveljavljena in dostopna Karl-Fischer metoda z opazovanjem spremembe barve pri končni točki titracije ni bila ustrezna, saj je bil izvleček obarvan. Zato smo razvili preprosto in finančno dostopno titracijsko metodo za ugotavljanje vsebnosti vode v obarvanih vzorcih, pri kateri končno točko titracije zaznamo spektrofotometrijsko. Tako ovrednoten izdelek predstavlja dobro osnovo za nadaljnje raziskovanje in razvijanje novih prehranskih dopolnil, kozmetičnih izdelkov in zdravil. Natural products are compounds or extracts that originate from natural sources and usually exhibit biological activity. They are raw materials for natural medicines. Pharmacognosy encompasses their selection, characterization, identification and use. Bioassay-guided fractionation of plant extracts linked to chromatographic separation techniques leads to the isolation of biologically active compounds whose chemical structures can readily be determined by modern spectroscopic and spectrometric methods, followed by testing of their pharmacological activity.Buckwheat contains many healthy nutrients, and its consumption is therefore increasing. The plant contains phototoxic compounds called fagopyrins that can act as photosensitisers upon excitation with visible light causing fagopyrism, but these compounds have received little research attention. To assure adequate safety of buckwheat and its products, fagopyrins should be characterized from a nutritional and pharmaceutical point of view. Nevertheless, fagopyrins are not only health-threatening compounds but also potent photodynamic agents that deserve further research. In this work, fagopyrins extraction procedure was improved employing a mixture of acetone and water. Flowers were recognized as the best material for the extraction. A chromatographic analysis of fagopyrins was performed by developing a simple, sensitive HPLC analytical method of high-resolution employing fluorescence detection. The developed method is a good starting point for a preparative method that would enable the isolation of pure compounds. At least eight fagopyrin derivatives were observed upon isolation and were characterized via UV-Vis absorption, NMR spectroscopy and mass spectrometry. The structures of two new fagopyrins were elucidated. Fagopyrin F is a previously known compound that consists of naphthodianthrone scaffold and two piperidine moieties. Fagopyrin A differentiates by two pyrrolidine moieties instead of piperidine moieties, and fagopyrin E consists of one pyrrolidine and one piperidine moiety. The existence of protofagopyrins that originate in buckwheat and can transform into fagopyrins upon light exposure was confirmed. Information on solubility, fluorescence and UV-Vis absorbance in different solvents were obtained. The need for reference compounds of different fagopyrins that would allow proper quantitative evaluation of buckwheat samples still remains. Researchers use hypericin for that purpose, but we recognize its use as inadequate due to the different structure and consequently uncomparable absorbance in spectrophotometric detection. Our methods complement the existing knowledge regarding fagopyrins and will facilitate their further isolation and analysis of their biological activity. In the second part of our work, silver fir trunk extracts were investigated. Extracts from the bark of different conifer species are known to contain various polyphenols that possess interesting pharmacological activities. So far the most extensive research was done on the antioxidative extract of the maritime pine (Pinus maritima) bark Pycnogenol®, which is widely used in food supplements and cosmetic products. We have shown that antioxidant activity of silver fir (Abies alba) extract is higher comparing to the maritime pine bark extract in vitro and in cultured cells. Components of the extract were separated with normal phase flash chromatography and reversed phase HPLC. Structures of individual compounds were elucidated by mass spectrometry, UV-Vis absorption spectroscopy and comparison to reference compounds. Six phenolic acids were identified (gallic, homovanillic, protocatehuic, p-hydroxybenzoic, vanillic and p-coumaric), three flavonoids (catechin, epicatechin and catechin tetramethyl eter) and four lignans (taxiresinol, 7-(2-methyl-3,4-dihydroxytetrahydropyran-5-yloxy)-taxiresinol, secoisolariciresinol and laricinresinol).The research was continued with pharmacological testing. It was found that the extract exerts strong antioxidative and protective effects against atherogenic diet-induced arterial wall damage. Protective effects of the silver fir extract and its compounds as well as of two phenolic acids (p-coumaric and protocatechuic acids) were also evauated against ischemia-reperfusion-induced damage in isolated rat hearts. The silver fir extract significantly reduced tissue damage (measured as the lactate dehydrogenase release rate) and shortened the duration of arrhythmias, which was not observed in the case of phenolic acids. It was concluded that the two acids are only partially responsible for the observed effects and synergism of more compounds could be present. Silver fir extract, solubilized in polyethylene glycol was developed as an appropriate raw material for research and use. A need for water determination in such samples emerged. The most accessible technique for water content determination is Karl-Fischer titration with visual detection of a titration endpoint, which was unfortunately not suitable for the analysis of the silver fir extract due to its strong colour. Therefore an appropriate method for moisture determination in coloured samples was developed employing simple equipment and spectrophotometric detection of a titration endpoint. Silver fir extract, characterized in such manner is a suitable ingredient for development of food supplements, cosmetic products and medicines.