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Biljke iz reda Brassicales poznate su kao dobar izvor glukozinolata i njihovih razgradnih produkata - izotiocijanata. U ovom istraživanju korišteni su uzorci eteričnih ulja i ekstrakata, dobivenih primjenom različitih metoda ekstrakcije, iz sjemenki gorušica te visokog dragoljuba. Budući da izotiocijanati imaju širok spektar bioloških aktivnosti od značaja je odrediti njihovu stabilnost i biodostupnost u uvjetima simuliranog procesa probave (želudac i tanko crijevo) koristeći ljudske i komercijalne probavne enzime. Glukozinolati i izotiocijanati iz navedenih uzoraka pokazali su veću gastrointestinalnu stabilnost nakon želučane faze probave u odnosu na crijevnu fazu. U ovom je istraživanju određena biološka aktivnost eteričnih ulja i ekstrakata smeđe i crne gorušice (antiproliferativna na dvije stanične linije karcinoma) i visokog dragoljuba (antiproliferativna na pet staničnih linija karcinoma, proapoptotska na dvije stanične linije karcinoma, vizualiziran je utjecaj uzoraka na stanice karcinom koristeći optičku i mikroskopiju atomskih sila – AFM te je određena antimikrobna aktivnost). Razgradni produkti glukozinolata prisutnih u gorušicama, pokazali su umjerenu i slabu antiproliferativnu aktivnost, dok razgradni produkti glukozinolata glukotropeolina prisutnog u visokom dragoljubu (benzil-izotiocijanat) su pokazali visok stupanj antiproliferativne, proapoptotske, antibakterijske i antiadhezijske aktivnosti. Plants of the order Brassicales are known as a good source of glucosinolates and their degradation products - isothiocyanates. In this work, samples of essential oils and extracts, from the mustard and nasturtium seeds were used, obtained by different extraction methods. Since isothiocyanates exhibit a wide range of biological activities, it is important to determine their stability and bioaccessibility under the conditions of a simulated digestion process (stomach and small intestine) with human and commercial digestive enzymes. The glucosinolates and isothiocyanates from the above samples showed higher gastrointestinal stability after the gastric phase of digestion compared to the intestinal phase. In this work, the biological activity of the essential oils and extracts of brown and black mustard (antiproliferative activity on two cancer cell lines) and nasturtium (antiproliferative activity on five cancer cell lines, proapoptotic activity on two cancer cell lines was determined, the effect of the essential oil on cancer cells was visualized by optical and atomic force microscopy - AFM, and the antimicrobial activity was determined). The degradation products of glucosinolates contained in mustard showed moderate and weak antiproliferative activity, while the degradation products of glucosinolate glucotropaeolin contained in nasturtium (benzyl isothiocyanate) showed a high level of antiproliferative, proapoptotic, antibacterial and antiadhesive activity.

Glukozinolati su produkti sekundarnog metabolizma identificirani u preko 16 različitih biljnih porodica od kojih je najznačajnija porodica Brassicaceae koja uključuje često korišteno povrće u ljudskoj prehrani. U biljkama se nalaze u odvojenim odjeljcima od enzima mirozinaze koja cijepa ove inače kemijski stabilne spojeve na razgradne produkte poput tiocijanata, izotiocijanata, nitrila i indola. Zbog zabilježenog biološkog djelovanja produkata razgradnje, predmet su mnogih istraživanja u posljednje vrijeme. Brassica oleracea var. italica, poznatija kao brokula, jedno je od najpopularnijih vrsta povrća. Hidroponski uzgojeno povrće u fazi klica za svakodnevnu konzumaciju sve je više u upotrebi. Predmet istraživanja ovoga radavsu klice brokule uzgajane u čistoj vodi te različitim koncentracijama selenija: 1 ppm, 2,5 ppm, 5 ppm, 7,5 ppm i 10 ppm s ciljem praćenja utjecaja na profil glukozinolata. Glukozinolati su izolirani, prevedeni u desulfo-oblik te analizirani UHPLC-DAD-MS/MS tehnikom. Analizom je utvrđena prisutnost 12 različitih glukozinolata: progoitrin, glukonapoleiferin, glukoalisin, glukonapin, glukokohlearin, 4-hidroksiglukobrasicin, glukobrasikanapin, glukobrasicin, glukoberteroin, glukonasturcin, 4-metoksiglukobrasicin i neoglukobrasicin. Uočena je dominantna količina glukonapina i glukobrasikanapina s maksimumima od 67,31 µmol/g i 36,14 µmol/g pri 5 ppm selenija. Glukoberteroin je uočen samo u tragovima. Povišene koncentracije selenija negativno su utjecale na rast brokule te su također utjecale na profil glukozinolata u različitim uzorcima. Dva uzorka klica brokule uzgajanih u koncentaciji selenija od 5 ppm i 7,5 ppm podvrgnuta su reakciji s enzimom mirozinazom da bi se dobili razgradni produkti glukozinolata koji su identificirani GC-MS-om. Identificirana su dva nitrila: benzenpropanonitril i 4,5-epitiopentanonitril. Drugi cilj eksperimenta je bio ispitati hoće li brokula kroz sedam dana apsorbirati dio selenija zbog kemijske sličnosti i obilja u kojem je rasla te ga ugraditi u strukturu svojih glukozinolata. Niti jedan selenoglukozinolat ili selenometionin-izvedeni glukozinolat nije detektiran niti su detektirani njihovi produkti razgradnje. Glucosinolates are products of secondary metabolism identified in over 16 different plant families, the most significant of which is the Brassicaceae family, which includes vegetables often used in human nutrition. In plants, glucosinolates are found in special compartments separated from the myrosinase enzyme, which cleaves these otherwise chemically stable compounds into degradation products such as thiocyanate, isothiocyanate, nitrile and indole. Due to the recorded biological activity of degradation products, they have been the subject of many recent researhces. Brassica oleracea var. italica, better known as broccoli, is one of the most popular types of vegetables. Hydroponically grown vegetables sprouts for everyday consumption are being increasingly used. In this work, broccoli sprouts were grown in clean water with different concentrations of selenium: 1 ppm, 2.5 ppm, 5 ppm, 7.5 ppm and 10 ppm to monitor its influence on the glucosinolate profile. Glucosinolates were isolated, converted into desulfo-form and analyzed by UHPLC-DAD-MS/MS technique. The analysis determined the presence of 12 different glucosinolates: progoitrin, gluconapoleiferin, glucoalisin, gluconapin, glucocochlearin, 4-hydroxyglucobrassicin, glucobrassicannapin, glucobrassicin, glucoberteroin, gluconasturtin, 4-methoxyglucobrassicin and neoglucobrassicin. A dominant amount of gluconapin and glucobrassicanapin was observed with maxima of 67.31µmol/g and 36.14 µmol/g, respectively, at a selenium concentration of 5ppm. Glucoberteroin was observed only in traces. Elevated concentrations of selenium had a negative effect on the growth of broccoli and also affected the profile of glucosinolates in different samples. Two samples of broccoli sprouts grown in selenium concentrations of 5 ppm and 7.5 ppm were subjected to reaction with the myrosinase enzyme to obtain glucosinolate degradation products that were identified by GC-MS. Two nitriles were identified: benzenepropanonitrile and cyano-3,4-epithiobutane. The second goal of the experiment was to examine whether the broccoli will absorb part of the selenium in seven days due to the chemical similarity and abundance in which it grew and incorporate it into the structure of its glucosinolates. No selenoglucosinolate or selenomethionine-derived glucosinolates were detected, nor their degradation products.

Glukozinolati su sumporom bogati specijalizirani biljni metaboliti zastupljeni u brojnim vrstama reda Capparales, gdje spadaju i prehrambeno značajne vrste porodice Brassicaceae (kupusnjače). Osnovna uloga glukozinolata u biljnom tkivu je obrana od herbivora i patogena. Glukozinolati su inertni sve dok ne dođu u kontakt s enzimom mirozinaza prilikom oštećenja tkiva. Mirozinaza hidrolizira glukozinolate u niz bioaktivnih, toksičnih produkata oporog okusa. Osim kod biotičkog stresa uzrokovanog djelovanjem biljojeda i patogena, glukozinolati sudjeluju i u regulaciji odgovora biljke na abiotičke stresore. Brojna istraživanja dokazala su pozitivan učinak redovitog konzumiranja kupusnjača na ljudsko zdravlje i neki produkti hidrolize glukozinolata posjeduju antikancerogena i protuupalna svojstva. Osim pozitivnog djelovanja na ljudsko zdravlje, pojedini glukozinolati i njihovi produkti razgradnje mogu imati i antinutritivna svojstva. Opisane su i bakterijske mirozinaze koje hidroliziraju glukozinolate u probavnom traktu sisavaca. Kuhanje i drugi procesi obrade biljaka, značajno utječu na bioraspoloživost glukozinolata. S obzirom na njihova ljekovita svojstva, istražuju se novi načini pohrane i obrade biljnog materijala kako bi se spriječio gubitak glukozinolata i maksimizirala bioraspoloživost. Brojna istraživanja posvećena su proučavaju različitih glukozinolata i njihovih produkata hidrolize u svrhu razvoja novih terapeutika. Glucosinolates are sulfur-rich specialized plant metabolites found in the order Brassicales. Glucosinolates play an important role in plant defense against herbivores and pathogens. Upon tissue damage, glucosinolates come into contact with enzyme myrosinase and are hydrolyzed to a variety of bioactive, toxic products of bitter taste. Glucosinolates also play an important role in regulating plant response to abiotic stress. Plenty of research shows that regular consumption of cruciferous vegetables has a positive impact on overall human health. Some products of glucosinolate degradation show anti inflammatory and anticancerogenic properties. There are also reports of antinutritive properties in some glucosinolate derivatives that can lead to a number of metabolic disorders. Other than plant myrosinase, bacterial myrosinases have also been described. Gut microbiota has an important role in glucosinolate hydrolysis in mammalian digestive system. Cooking and other forms of plant tissue processing have a huge impact on glucosinolate availability. Considering the medicinal properties of glucosinolates, there is a lot of effort given in optimizing the existing and finding new methods of storing and processing of vegetables to maximize the bioavailability. Much research is focused in developing new therapeutics based on glucosinolates or their derivatives to fight diseases.

SAŽETAK Raspodjela glukozinolata unutar biljnih porodica reda Brassicales istražena je UHPLC- DAD-MS/MS analizom desulfoglukozinolata na temelju određenih kvalitativnih i kvantitativnih informacija i to u različitim biljnim tkivima četrdeset osam (48) biljaka iz pet (5) biljnih porodica (Brassicaceae, Capparidaceae, Moringaceae, Resedaceae, Tropaeolaceae) koje rastu u Republici Hrvatskoj i šire. Razvijena je baza MS2 spektara desulfoglukozinolata korištenjem komercijalno dostupnih standarda kao i glukozinolata izoliranih iz prirodnih izvora. Biljna porodica Brassicaceae je sadržavala najraznolikije glukozinolate koje potječu od sedam (7) aminokiselinskih prekursora, i to najčešće od metionina, praćeno s glukozinolatima koji potječu od triptopfana te fenilalanina i/ili tirozina, dok su razgranati alifatski glukozinolati koji potječu od valina, leucina i izoleucina zastupljeni tek u rijetkim vrstama. Jedini poznati glukozinolat, glukokaparin, koji potječe od aminokiseline alanina identificiran je samo u porodici Capparidaceae i to kao glavni u svim istraženim vrstama, dok su ostali glukozinolati u većoj mjeri potjecali od triptofana. Moringa oleifera (porodica Moringaceae) sadržavala je 4-(α-L- ramnopiranoziloksi)benzil-glukozinolat te u manjim količinama tri (3) O-acetilirana izomera istog glukozinolata čiji aminokiselinski prekursor je fenilalanin i/ili tirozin. U vrstama Reseda lutea i R. phyteuma (porodica Resedaceae) identificiran je izomer 4-(α-L-ramnopiranoziloksi)benzil-glukozinolata, 2-(α-L-ramnopiranoziloksi)benzil- glukozinolat karakterističan za biljnu porodicu Resedaceae, koji potječe od fenilalanina, dok je u R. alba kao glavni glukozinolat identificiran 2-hidroksi-2-metilpropil- glukozinolat koji potječe od izoleucina. Tropaeolum majus je sadržavao arilalifatski benzil-glukozinolat kao glavni, praćen sa 3-metoksibenzil-glukozinolatom, a oba potječu od fenilalanina, jedinog poznatog aminokiselinskog prekursora glukozinolata u porodici Tropaeolaceae. Uvidom u kvantitativni sastav glukozinolata odabrane su biljne vrste kao dobri izvori glukozinolata koji su izolirani u obliku desulfoglukozinolata i potvrđeni spektroskopskim tehnikama (MS2, 1 H, 13 C, COSY i HSQC NMR). 3-(Metilsulfinil)propil-glukozinolat je izoliran iz biljke Anastatica hierochuntica (porodica Brassicaceae), 4-hidroksibenzil-glukozinolat iz vrste Sinapis alba (porodica Brassicaceae), metil-glukozinolat iz Capparis orientalis (porodica Capparidaceae), 2-hidroksi-2-metilpropil-glukozinolat iz biljke Reseda alba (porodica Resedaceae) te benzil-glukozinolat iz Tropaeolum majus (porodica Tropaeolaceae). Također, na temelju MS2 spektara i biosintetskih karakteristika identificirana su 4 nova glukozinolata: orto ili meta hidroksi-4-(α-L-ramnopiranoziloksi)benzil-glukozinolat (Moringa oleifera), 4´-O- (β-D-apiofuranoziloksi)benzil-glukozinolat (Hesperis laciniata i Thlaspi perfoliatum), 4- hidroksi-1-metoksiindol-3-ilmetil-glukozinolat (Thlaspi perfoliatum) te 3-hidroksi-(4- metilsulfonil)butil-glukozinolat (Barbarea vulgaris) čije strukture trebaju dodatnu potvrdu NMR-om. Stabilnost desulfo-4-(metilsulfinil)but-3-enil-glukozinolata izoliranog iz biljke Matthiola incana ispitivana je tijekom jednomjesečnog razdoblja te je utvrđena njegova nestabilnost u vodenoj otopini, odnosno drastičan pad sadržaja (do 80 %) praćen spontanom konverzijom u cikličke tioimidatne N-okside. Korištenjem konvencionalnih (enzimska hidroliza i hidrodestilacija) te modernih metoda (mikrovalna destilacija te hidrodifuzija i gravitacija) ispitan je enzimski i termički utjecaj na razgradnju glukozinolata. U uvjetima termičke razgradnje potpomognute mikrovalovima dobiveni su manji prinosi razgradnih produkata u odnosu na termičku razgradnju tijekom hidrodestilacije te enzimsku hidrolizu. Izotiocijanati kao razgradni produkti glukozinolata su se pokazali nestabilnim pri uvjetima GC-MS mjerenja i to za ω-metilsulfinilne C3, C5 i C6 izotiocijanate čiji je raspad rezultirao nastankom odgovarajućih olefinskih izotiocijanata. Dodatno, zbog prisutnosti elektron-donirajuće hidroksilne skupine u para položaju na benzenskom prstenu, kod 4-hidroksibenzil- izotiocijanata i 4-hidroksi-3,5-dimetoksibenzil-izotiocijanata, utvrđena je nestabilnost koja vodi ka nastanku artefakata tijekom GC-MS mjerenja. Također, razgradni produkti indolnih glukozinolata nisu identificirani. Ovime je istaknut nedostatak neizravne metode identifikacije glukozinolata preko razgradnih produkata u biljnim vrstama. U biljci Eruca vesicaria biofortificiranoj selenijem iz natrijevog selenata identificiran je 4-(metilseleno)butil-glukozinolat izveden iz selenometionina na temelju MS2 spektra koji je po prvi puta prikazan. Utvrđeno je da hidroponski uzgojene klice pet (5) odabranih biljaka iz porodice Brassicaceae imaju visoki sadržaj karakterističnih glukozinolata zbog čega predstavljaju dobar izvor za njihovu izolaciju. ABSTRACT Distribution of glucosinolates within plant families of the order Brassicales was investigated using qualitative and quantitative UHPLC-DAD-MS/MS analysis of desulfoglucosinolates in different plant tissues of forty-eight (48) plants from five (5) plant families (Brassicaceae, Capparidaceceae, Moringaceae, Resedaceae, Tropaeolaceae) growing in the Republic of Croatia and worldwide. A database of MS2 spectra of desulfoglucosinolates was developed using commercially available standards as well as glucosinolates isolated from natural sources. The Brassicaceae plant family contained a wide variety of glucosinolates derived from seven (7) different amino acid precursors, most commonly from methionine, followed by glucosinolates derived from tryptophan, phenylalanine and/or tyrosine, while branched aliphatic glucosinolates deriving from valine, leucine and isoleucine were scarce. Glucocapparin, the only known glucosinolate derived from the amino acid alanine, was identified solely in the Capparidaceae family as the main one in all the species studied, while the other glucosinolates largely derived from tryptophan. Moringa oleifera (family Moringaceae) contained 4-(α-L-rhamnopyranosyloxy)benzyl glucosinolate and, in less amount, three O-acetylated isomers of the same glucosinolate derived from phenylalanine and/or tyrosine. In the species Reseda lutea and R. phyteuma (family Resedaceae) an isomer of 4-(α-L-rhamnopyranosyloxy)benzyl glucosinolate, 2- (α-L-rhamnopyranosyloxy)benzyl glucosinolate, was identified characteristic for the plant family Resedaceae, originating from phenylalanine. On the other hand, 2-hydroxy- 2-methylpropyl glucosinolate derived from isoleucine was the major glucosinolate identified in R. alba. The main glucosinolate in Tropaeolum majus was arylaliphatic benzyl glucosinolate followed by 3-methoxybenzyl glucosinolate, both derived from phenylalanine, the only known amino acid precursor of glucosinolates in the Tropaeolaceae family. Insight into the quantitative composition of glucosinolates enabled the selection of the plant species as the sources of glucosinolates for their isolation in the form of desulfoglucosinolates and confirmation by spectroscopic techniques (MS2, 1 H, 13 C, COSY and HSQC NMR). 3-(Methylsulfinyl)propyl glucosinolate was isolated from Anastatica hierochuntica (family Brassicaceae), 4-hydroxybenzyl glucosinolate from Sinapis alba (family Brassicaceae), methyl glucosinolate from Capparis orientalis (family Capparidaceae), 2-hydroxy-2-methylpropyl glucosinolate from Reseda alba (family Resedaceae) and benzyl glucosinolate from Tropaeolum majus (family Tropaeolaceae). Also, based on MS2 spectra and biosynthetic characteristics, four (4) new glucosinolates were identified: ortho or meta hydroxy-4-(α-L- rhamnopyranosyloxy)benzyl glucosinolate (Moringa oleifera), 4´-O-(β-D- apiofuranosyloxy)benzyl glucosinolate (Hesperis laciniata and Thlaspi perfoliatum), 4- hydroxy-1-methoxyindol-3-ylmethyl glucosinolate (Thlaspi perfoliatum) and 3-hydroxy- (4-methylsulfonyl)butyl glucosinolate (Barbarea vulgaris) whose structures need further confirmation by NMR. The stability of desulfo-4-(methylsulfinyl)but-3-enyl glucosinolate isolated from Matthiola incana was studied over a one-month period and its instability in aqueous solution was observed, showing a significant drop in the content (up to 80%) followed by spontaneous conversion to cyclic thioimidate N-oxides. The enzymatic and thermal degradation of glucosinolates was studied by using conventional (enzymatic hydrolysis and hydrodistillation) and modern methods (microwave distillation and hydrodiffusion and gravity). Under microwave-assisted thermal decomposition conditions, lower yields of decomposition products were obtained compared to thermal decomposition during hydrodistillation and enzymatic hydrolysis. Isothiocyanates as degradation products of glucosinolates showed to be unstable at GC-MS conditions for ω-methylsulfinyl C3, C5 and C6 isothiocyanates, which decomposition resulted in the formation of the corresponding olefinic isothiocyanates. Additionally, due to the presence of an electron- donating hydroxyl group in the para position on the benzene ring, in 4-hydroxybenzyl isothiocyanate and 4-hydroxy-3,5-dimethoxybenzyl isothiocyanate, instability lead to formation of artifacts found during GC-MS analysis. Also, degradation products of indole glucosinolates have not been identified. This highlights the disadvantage of an indirect method for the identification of glucosinolates via their degradation products in plant species. In the Eruca vesicaria biofortified by selenium from sodium selenate, 4- (methylseleno)butyl glucosinolate derived from selenomethionine was identified based on the MS2 spectrum shown for the first time. Hydroponically grown sprouts of the five (5) selected plants of Brassicaceae family were found to have a high content of characteristic glucosinolates presenting a good source for their isolation.

Raštika (Brassica oleracea var. acephala L.) pripada porodici krstašica (kupusnjača) (lat. Brassicaceae) koje se od davnina koriste u ljudskoj prehrani, a poznat su izvor biološki aktivnih spojeva. Istraživanja su pokazala da raštika, kao i ostale vrste porodice krstašica, obiluje sumporom bogatim spojevima glukozinolatima, koji povoljno djeluju na ljudsko zdravlje. Cilj ovog istraživanja bio je ispitati citotoksično djelovanje triju glukozinolata (sinigrina, glukoiberina i progoitrina) izoliranih iz raštike na stanice raka debelog crijeva (HCT116). Sva tri izolirana glukozinolata uspješno su zaustavila rast tumorskih HCT116 stanica. Najveću aktivnost imao je glukoiberin (\(IC_{50}\) = 51,67 μg/mL), upola slabiju aktivnosti progoitrin (\(IC_{50}\) = 156,31 μg/mL), a najmanju sinigrin (\(IC_{50}\) = 1128,67 μg/mL). Rezultati ukazuju na to da testirani glukozinolati imaju značajan citotoksični potencijal što otvara mogućnost daljnjeg istraživanja njihove biološke aktivnosti i mogućeg korištenja u razvoju novih vrsta prirodnih kemoterapeutskih spojeva., Borecole (Brassica oleracea var. acephala L.) belongs to the cruciferous family (lat. Brassicaceae), which has been used in human nutrition since ancient times and is a well-known source of biologically active compounds. Research has shown that borecole, like other species of the cruciferous family, is rich in sulfur-containing glucosinolate compounds that have beneficial effects on human health. The aim of this study was to investigate the cytotoxic effect of three glucosinolates (sinigrine, glucoiberine, and progoitrine) isolated from borecole on colon cancer cells (HCT116). All three isolated glucosinolates successfully stopped the growth of HCT116 tumor cells. Glucoiberine had the highest activity (\(IC_{50}\) = 51.67 μg/mL), progoitrine (\(IC_{50}\) = 156.31 μg/mL) had half the activity, and sinigrine (\(IC_{50}\) = 1128.67 μg/mL) had the lowest activity. The results suggest that the tested glucosinolates have a significant cytotoxic potential, which opens the possibility of further exploring of their biological activity and using them in the development of new types of natural chemotherapeutic compounds.

Biljka Lepidium sativum L. je jednogodišnja kultura koja se na zapadu često koristi u ljudskoj prehrani radi dobrih svojstava, izdržljivosti i lakog uzgoja. U ovom radu je prikazan uzgoj klica biljke Lepidium sativum L. na vodenim otopinama željeza različitih koncentracija te je praćen njihov razvoj tijekom dva tjedna, a zatim je prikazana metoda pripreme biljnog materijala za analizu atomskim apsorpcijskim spektrometrom. Koristeći se navedenom tehnikom, određena je koncentracija željeza u klicama. Uočen je porast koncentracije željeza u uzgajanim klicama, u skladu s povećanjem koncentracije željeza u vodenim otopinama na kojima su biljke uzgajane. U ovome radu izolirani su glukozinolati iz sjemena uzgojenih klica Lepidium sativum L. metodom ekstrakcije 70 %-tnim metanolom uz desulfataciju enzimom sulfataza te određeni UHPLC-DAD-MS/MS tehnikom. U uzorku klica obogaćenih vodenom otopinom željeza od 1 ppm u odnosu na referentni uzorak uočava se pad koncentracije glukotropeolina i povećanje 4-metoksiglukobrasicina. Nadalje, kod klica koje su uzgajane na vodenim otopinama željeza koncentracije 5 ppm i 10 ppm uočen je pad glukotropeolina kao i pad koncentracije 4-metoksiglukobrasicina. Kod klica uzgajanim na vodenoj otopini željeza koncentracije 25 ppm uočen je pad koncentracije glukotropeolina u odnosu na referentni uzorak kao i povećanje koncentracije 4-metoksiglukobrasicina. The plant Lepidium sativum L. is an annual crop that is often used in the human diet in the West for its good properties, durability and ease of cultivation. In this paper, the cultivation of Lepidium sativum L. bean sprouts on aqueous solutions of iron of different concentrations is presented and their development is monitored for two weeks. The method of preparation of plant material for analysis by atomic absorption spectrometer is presented. Using the above mentioned technique the concentration of iron in the bean sprouts was determined. An increase in the concentration of iron in the grown bean sprouts was observed, in line with the increase in the concentration of iron in the aqueous solutions on which the plants were grown. In this work, glucosinolates were isolated from the seeds of cultivated bean sprouts of Lepidium sativum L. by the method of extraction with 70% methanol with desulfation with the enzyme sulfatase and determined by UHPLC-DAD-MS / MS technique. In a sample of bean sprouts grown on an aqueous solution of iron of 1 ppm compared to the reference sample, a decrease in the concentration of glucotropeolin and an increase in 4-methoxyglucobrasicin was observed. Furthermore, in bean sprouts grown on aqueous iron solutions with concentrations of 5 ppm and 10 ppm, a decrease in glucotropeolin as well as a decrease in the concentration of 4- methoxyglucobrasicin was observed. In bean sprouts grown on aqueous iron solution with a concentration of 25 ppm, a decrease in the concentration of glucotropeolin compared to the reference sample was observed, as well as an increase in the concentration of 4-methoxyglucobrasicin.

Glukozinolati su sekundarni metaboliti koji se sintetiziraju iz aminokiselina. Identificirani su u preko 16 biljnih porodica, a najznačajnija je porodica Brassicaceae u koju se ubraja biljka Lepidium sativum L. Glukozinolati su kemijski i biološki neaktivni spojevi te im je zajedničko da su po kemijskoj strukturi β- tioglukozidni-N-hidroksisulfati s varijabilnim bočnim lancem. Kres salata (Lepidium sativum L.) je jednogodišnja kultura koja sadrži mnoge ljekovite tvari, ali je u ljudskoj prehrani slabo rasprostranjena. Navodi se da je kres salata dobar hiperakumulator teških metala. Biljke nemaju potrebu za kadmijem i uglavnom ne podnose koncentracije veće od 1 ppm, no navedena biljka može akumulirati kadmij u tkivima i tolerirati koncentraciju do 100 ppm. U ovom radu opisan je uzgoj biljke Lepidium sativum L. koji je zalijevan s otopinama kadmija različitih koncentracija od 1 do 50 ppm. Razvoj klica praćen je tijekom 7 dana i jedino pri koncentraciji od 50 ppm došlo je do propadanja biljke. Izolacija glukozinolata se vršila ekstrakcijom uz pomoć 70%-tnog metanola uz desulfataciju, dok se analiza vršila vezanom tehnikom UHPLC-DAD-MS/MS. Identificirani su sljedeći glukozinolati: glukotropeolin koji se nalazi u svim uzorcima i 4-metoksiglukobrasicin koji nije pronađen jedino u sjemenu. Glukotropeolin spada u skupinu arilalifatskih, dok 4-metoksiglukobrasicin pripada skupinu heterocikličnih (indolnih) glukozinolata. Sadržaj glukozinolata, s obzirom na to da su se uzorci zalijevali otopinama kadmija različitih koncentracija, nije se značajno mijenjao. U radu je također opisana priprema biljnog materijala za analizu atomskom apsorpcijskom spektroskopijom. Prije početka analiziranja potrebno je napraviti krivulju umjeravanja, zato što se preko nje iščitavaju koncentracije kadmija u biljnom materijalnu. Na temelju dobivenih rezultata izvodi se zaključak da li se biljka Lepidium sativum L. može koristiti kao hiperakumulator kadmija. Glucosinolates are secondary metabolites that are synthesized from amino acids. They were found in over 16 plant families, the most important being the Brassicaceae family, which includes the plant Lepidium sativum L. Glucosinolates are chemically and biologically inactive compounds and have in common that they are chemically β-thioglucoside-N-hydroxysulfates with a variable side chain. The Lepidium sativum L. plant is an annual crop that contains many medicinal substances, but is poorly used in the human diet. Cress salad is known to be a good hyperaccumulator of heavy metals. Plants do not need cadmium and generally do not tolerate concentrations higher than 1 ppm, but this plant can accumulate cadmium in tissues and tolerate concentrations up to 100 ppm. This thesis describes the cultivation of the plant Lepidium sativum L., which was watered with cadmium solutions of different concentrations from 1 to 50 ppm. Sprouts development was monitored for 7 days and only at a concentration of 50 ppm did the plant decay. Isolation of glucosinolate was performed by extraction with 70% methanol with desulfation, while the analysis was performed using UHPLC-DAD- MS/MS. The following glucosinolates were identified: glucotropeolin was found in all samples and 4- methoxyglucobrasicin which was not found only in seeds. Glucotropeolin belongs to the group of arylaliphatic, while 4-methoxyglucobrasicin belongs to the group of heterocyclic (indole) glucosinolates. The glucosinolate content, since the samples were watered with cadmium solutions of different concentrations, did not change significantly. The thesis also describes the preparation of plant material for analysis by atomic absorption spectroscopy. Before starting the analysis, it is necessary to make a calibration curve because the cadmium concentrations in the plant material are read through it. Based on the obtained results, it was concluded that the plant Lepidium sativum L. can be used as a hyperaccumulator of cadmium.

Cilj istraživanja bilo je utvrditi stabilnost sinigrina i sinalbina u dvofaznom simuliranom probavnom sustavu. Ispitivani su sinalbin iz bijele gorušice (Sinapis alba L.) i sinigrin iz smeđe gorušice (Brassica juncea L.) koristeći metodu simulacije želučane i crijevne faze. Daljni cilj bio je usporediti stabilnost glukozinolata u odnosu na komercijalne i ljudske enzime u svakoj od dviju faza probave. Za simuliranje ljudske želučane i crijevne faze korišteni su ljudski želučani i crijevni sokovi skupljeni u Kliničkom bolničkom centru Split, a kao komercijalni enzimi korištene su smjese SGF (eng. simulated gastric fluid) za želučanu i SIF (eng. simulated intestinal fluid) za crijevnu fazu. Rezultati su obrađivani na spojenom sustavu HPLC-DAD-MS/MS. Veća stabilnost dokazana je u želučanoj fazi u odnosu na crijevnu fazu za oba glukozinolata. Nadalje, veća stabilnost uočena je prilikom korištenja ljudskih probavnih sokova u usporedbi s komercijalnim smjesama enzima, također za oba glukozinolata i u objema fazama probave. Izlaganje glukozinolata, u ovom slučaju sinigrina i sinalbina, probavnim sokovima dovodi do njihove degradacije i nastajanja razgradnih produkata izotiocijanata. Glukozinolati su podložni razgradnji i u želučanoj i u crijevnoj fazi probave, no veća razdradnja zbiva se u crijevima. Nadalje, komercijalni enzimi u većoj mjeri razgrađuju glukozinolate od ljudskih probavnih enzima, no u obzir se mora uzeti velika interindividualna varijabilnost između ljudskih probavnih sokova., The aim was to determine the stability of sinigrin and sinalbin in a two-phase simulated digestive system. Sinalbin from white mustard (Sinapis alba L.) and sinigrin from yellow mustard (Brassica juncea L.) were studied using a method that simulates the gastric and intestinal phases. Another objective was to compare the stability of glucosinolates in relation to human and commercial enzymes in each of the two digestion phases. To simulate the human gastric and intestinal phases, human gastric and intestinal juices collected from the University Hospital of Split were used. SGF (simulated gastric fluid) for gastric phase and SIF (simulated intestinal fluid) for intestinal phase were used as commercial mixtures. The results were analyzed using HPLC-DAD-MS/MS system. Higher stability was demonstrated in the gastric phase compared to the intestinal phase. Moreover, when human digestive juices were used, higher stability was observed compared to commercial enzyme mixtures for both glucosinolates and in both digestive phases of digestion. When glucosinolates, in this case sinigrin and sinalbin, are exposed to digestive juices, it leads to their degredation and the formation of their degradation products isothiocyanates. Glucosinolates can be degraded in both the gastric and intestinal phase, with degradation being greater in the intestine. In addition, commercial enzymes degrade glucosinolates more than human digestive enzymes, although the large interindividual variability between human digestive juices must be taken into account.

Glukozinolati (β-tioglukozidni-N-hidroksisulfati) su velika grupa sekundarnih metabolita koji daju karakterističan gorki i oštri okus kupusnjačama te su njihovi razgradni produkti biološki aktivni spojevi. Razgradnja može biti termička, enzimska ili kemijska pri čemu nastaju brojni razgradni produkti kao što su tiocijanati i izotiocijanati. Klice rikule su nutritivno bogate, a prednost im je vrlo laki uzgoj. Klijanjem se nutrijenti u sjemenci razgrađuju na sastavne dijelove, razvija se klorofil i povećava se količina proteina, minerala i elemenata u tragovima. Dodatna prednost je da su bogate vitaminima, čiji se sadržaj povećava klijanjem, esencijalnim masnim kiselinama i vlaknima te imaju pozitivan učinak na organizam. Biljke nemaju potrebu za selenom i uglavnom ne podnose koncentracije veće od 10-100 μg/g suhe mase u tkivima, no vrste koje pripadaju obitelji Brassicaceae mogu akumulirati selen u tkivima i tolerirati koncentracije do 1000 μg/g suhe mase. Na taj način, biljke proizvode razne seleno metabolite koji mogu imati ulogu kao sredstva za sprječavanje raka. Istraživanja su pokazala da se u bočni lanac glukozinolata umjesto sumpora može ugraditi atom selena. U ovom diplomskom radu uzgojene su klice rikule te zalijevane otopinama selena različitih koncentracija (1; 2,5; 5; 7,5; 10 ppm). Razvoj biljaka praćen je tijekom 7 dana i jedino pri najvišoj koncentraciji selena nije došlo do razvoja biljke. Nakon sabiranja glukozinolati su izolirani ekstrakcijom 70%-tnim metanolom te analizirani UHPLC-DAD-MS/MS, a njihovi hlapljivi razgradni produkti su izolirani ekstrakcijom diklormetanom i identificirani GC-MS tehnikom. Identificirana su 4 glukozinolata: glukorafanin, glukoerucin, dimer glukosativina i 4-metoksiglukobrasicin, dok je GC-MS tehnikom identificiran erucin kao razgradni produkt glukoerucina. Glucosinolates (β-thioglucoside-N-hydroxysulfates) are a large group of secondary metabolites that give a characteristic bitter and pungent taste to cabbages and their degradation products are biologically active compounds. Degradation can be thermal, enzymatic, or chemical and it forms several degradation products such as thiocyanates and isothiocyanates. Rocket sprouts are nutritionally rich, and their advantage is very easy cultivation. By germination, the nutrients in the seed are broken down into their main components, chlorophyll develops and the amount of proteins, minerals and trace elements increases. An additional advantage is that they are rich in vitamins whose content increases with germination, essential fatty acids and fiber and have a positive effect on the body. Plants do not need selenium and generally do not tolerate concentrations higher than 10-100 μg/g dry weight in tissues, but species belonging to the Brassicaceae family can accumulate selenium in tissues and tolerate concentrations up to 1000 μg/g dry weight. In this way, plants produce various selenium metabolites that can play a role as a means of preventing cancer. Studies have shown that a selenium atom can be incorporated into the side chain of glucosinolates instead of sulfur. In this diploma thesis, rocket sprouts were grown and watered with selenium solutions of different concentrations (1; 2.5; 5; 7.5; 10 ppm). The development of the plants was monitored for 7 days, and only at the highest concentration of selenium the plant did not develop. After collection, glucosinolates were isolated by extraction with 70% methanol and analyzed by UHPLC-DAD-MS / MS, and their volatile degradation products were isolated by extraction with dichloromethane and identified by GC-MS technique. 4 glucosinolates were identified: glucoraphanin, glucoerucine, glucosatin dimer and 4-methoxyglucobrasicin, while GC-MS technique identified erucin which originate from degradation of glucoerucin.

Glukozinolati su sekundarni biljni metaboliti β-tioglukozidne strukture koji hrani biljnog podrijetla daju aromu, miris i okus. Smatraju se fitokemikalijama što znači da imaju potencijal u prevenciji različitih bolesti u ljudskom organizmu, a naročito pojave karcinoma. U ovom radu provela se identifikacija glukozinolata, te kvantizacija glukozinolata u prahu žutog i crvenog korijena Mace te Moringa praha i čaja. Lepidium meyenii (Brassicaceae) - Maca je bogata hranjivim elementima velike vrijednosti i sekundarnim metabolitima visoke biološke aktivnosti. U ovom radu određen je glukozinolatni profil iz dvije biljne vrste L. meyenii, žute i crvene. Kromatogram potvrđuje činjenicu da su prisutni slični glukozinolati. Glukotropeolin (benzil-glukozinolat) je u oba uzorka glavni pri čemu njegova količina u žutoj iznosi 7,02 μmol/g, a u crvenoj 8,96 μmol/g suhog biljnog materijala. Glukotropeolin je praćen s glukolimnantinom (3-metoksibenzil-glukozinolat) čija koncentracija u žutoj iznosi 1,93 μmol/g, a u crvenoj 2,55 μmol/g suhog biljnog materijala. Moringa oleifera (Moringaceae), poznata kao Moringa, podrijetlom je iz sjeverozapada Indije. U Africi se često konzumira za samo-liječenje bolesnika koji su pogođeni dijabetesom, hipertenzijom ili HIV / AIDS-om. U Moringi kao glavni spoj izdvaja se glukomoringin (4-(α-L-ramnoziloksi)benzil-glukozinolat, zastupljen u čaju u količini od 29,23 μmol/g i prahu 21,78 μmol/g suhog biljnog materijala. Acetilirani izomeri glukomoringina u Moringi su zastupljeni u znatno manjim količinama. Glucosinolates are plant secondary metabolites of β-thioglucoside structure that are responsible for flavour, aroma and taste of foods of plant origin. They are considered phytochemicals which means that they have the potential to protect the human body from various diseases, especially with the onset of cancer. In this work, the identification of glucosinolates and the quantization of glucosinolates in the powder of yellow and red roots of Maca and Moringa powder and tea were examined. Lepidium meyenii (Brassicaceae) - Maca is rich in nutrients of high value and secondary metabolites with high biological activities. In this paper, the glucosinolate profile from two plant species Lepidium meyenii (Maca), yellow and red, is correct. The chromatogram exposes the fact that similar glucosinolates are present. Glucotropeolin (benzyl glucosinolate) is the main glucosinolate in both samples having 7.02 μmol/g of dry weight in yellow Maca powder, and 8.96 μmol/g of dry weight in red Maca powder. Glucotropeolin was followed by glucolimnanthine (3-methoxybenzyl-glucosinolate), having 1.93 μmol/g of yellow Maca powder and 2.55 μmol/g of red Maca powder. Moringa oleifera (Moringaceae), commonly known as Moringa, is native to north-western India. In Africa, it is often consumed for the self-treatment of patients affected by diabetes, hypertension or HIV / AIDS. Glucomoringin (4- (α-L-ramnosyloxy)benzyl glucosinolate was identified as the main compound in Moringa, with 29.23 μmol/g of dry tea weight and 21.78 μmol/g of Moringa powder. The acetylated isomers of glucomoringin in both Moringa samples were identified in much smaller amounts.

Glukozinolati su sekundarni metaboliti identificirani u preko 16 različitih biljnih porodica, a najznačajnija je porodica Brassicaceae u koju se ubrajaju dobro poznate biljke iz svakodnevne prehrane. Glukozinolati su fiziološki neaktivni i kemijski stabilni i tek njihovom razgradnjom djelovanjem enzima mirozinaze dolazi do oslobađanja raznovrsnih spojeva. Smatraju se fitokemikalijama s potencijalom prevencije različitih bolesti u ljudskom organizmu. U ovom diplomskom radu izolirani su i analizirani glukozinolati iz odabranih dijelova biljaka Nasturtium officinale i Iberis umbellata. Glukozinolati su identificirani UHPLC-DAD-MS/MS analizom desulfatiranih glukozinolata. Isti su kvantificirani u različitim dijelovima biljke prema službenoj ISO 9167-1 metodi koja se zasiva na HPLC analizi desulfoglukozinolata. Izolacija glukozinolata iz navedenih biljnih materijala je napravljena metodom ekstrakcije glukozinolata uz desulfataciju. Kod Nasturtium officinale izolirani su i kvantizirani sljedeći glukozinolati: metilsulfinilalkilni glukozinolati i to glukohesperin, 7-(metilsulfinil)heptil-glukozinolat, glukohirsutin, zatim glukonasturcin koji je aril-alkilni glukozinolat te glukobrasicin i 4-metoksiglukobrasicin koji spadaju u skupinu indolnih glukozinolata. U sjemenu Iberis umbellata kvantizirani su spojevi: glukoiberin, sinigrin, glukonapin, glukoibervirin i glukotropeolin, dok su iz tog sjemena uzgojenoj biljci Iberis umbellata identificirani sljedeći spojevi: glukoiberin, sinigrin, glukoibervirin, glukotropeolin i 4-metoksiglukobrasicin. U uzgojenoj biljci Iberis umbellata nije prisutan glukonapin kojeg nalazimo u sjemenu, ali je identificiran 4-metoksiglukobrasicin koji nije pronađen u sjemenu. Glucosinolates are secondary metabolites identified in over 16 different plant families, among which the most important is Brassicaceae family, which includes well known plants of everyday nutrition. Glucosinolates are physiologically inert and chemically stable upon activation of the enzyme myrosinase leads to the degradation of glucosinolates and formation of various compounds. They are considered as phytochemicals which have potential to protect the human body form various diseases. In this diploma thesis, glucosinolates from selected parts of Nasturtium officinale and Iberis umbellata plants were isolated and analysed. Glucosinolates were identified by UHPLC-DAD-MS/MS analyzing desulfated glucosinolates. The same were quantified in different plant parts according to the official ISO 9167-1 method based on the HPLC analysis of desulphoglucosinolates. Isolation of glucosinolates from used plant material was performed using the method of glucosinolate extraction by desulphation. The following glucosinolates were isolated and quantizated from Nasturtium officinale: methylsulfinylalkyl glucosinolates i.e. glucohesperin, 7-(methylsulfinyl)heptylglucosinolate, glucohirsutin, than gluconasturcin which is arylalkyl glucosinolate, and glucobrasicin and 4-methoxyglucobrasicin which belong indole glucosinolates group. The following compounds were quantizated in Iberis umbellata seeds: glucoiberin, sinigrin, gluconapin, glucoibervirin and glucotropeolin. While from Iberis umbellata plant cultivated using the same seed following compounds were indentified: glucoiberin, sinigrin, glucoibervirin, glucotropeolin and 4-methoxyglucobrasicin. Although present in seeds, gluconapin was not found in cultivated Iberis umbellata plant; on the other hand 4-methoxyglucobrasicin was identified in the plant whie beeing absent in the seeds.

Duboka, vlažna ilovasta tla s dobrim drenažnim sustavom i umjerenom klimom najpovoljnija su za uzgoj jednogodišnjeg i višegodišnjeg hrena, a zemlje koje su najveći uzgajivači hrena su Sjedinjene Američke Države i Mađarska. Hren se zbog svojega specifičnoga okusa koristi u prehrani kao začin te ima višestruko zdravstveno djelovanje zbog velike količine vitamina C te zbog antibiotskih svojstava. Važna je sirovina za izdvajanje peroksidaze koja se koristi za reagense pri određivanju šećera i kolesterola u krvi. Hren je bogat glukozinolatima, sekundarnim produktima koji imaju važnu ulogu u obrambenom sustavu biljke. Uz glukozinolate, važno je istaknuti i izotiocijanat koji bi mogao postati važno antikancerogeno sredstvo jer, osim što sprječava razvoj novih, uništava postojeće kancerogene stanice. Uz antibakterijske, izotiocijanat posjeduje i antiprototozoalne i antifungalne aktivnosti. Upravo je zbog hranjivih vrijednosti, ljekovitih svojstava i specifičnog okusa i mirisa preporučljiva konzumacija hrena u različitim oblicima., Deep, moist, loamy soils with good drainage systems and temperate climates are best suited for growing horseradish and the countries that are the largest horseradish producers are the United States of America and Hungary. Due to its specific taste, horseradish is used in the diet as a spice and has multiple health effects due to its high amount of vitamin C and its antibiotic properties. An important raw material obtained from horseradish is peroxidase which is used for tests in the determination of blood sugar and cholesterol. Horseradish is rich in glucosinolates, secondary products that play an important role in the plant's defense system. In addition to glucosinolates, it is also important to highlight isothiocyanate, which could become an important anticarcinogenic agent because, in addition to preventing the development of new ones, it destroys existing cancer cells. In addition to antibacterials, isothiocyanate also possess antiprotozoal and antifungal activities. Due to its nutritional value, medicinal properties, specific taste and aroma, it is advisable to consume horseradish in various forms.

U radu je prikazan doprinos istraživanju glukozinolata u razdoblju 2005. – 2020. koji je ostvaren radom na Kemijsko-tehnološkom fakultetu, Sveučilišta u Splitu. Pregledom sveoubuhvatne dokumentacije utvrđeno je da znatnom broju predloženih struktura nedostaju važne strukturne informacije koje bi osigurala NMR i MS spektroskopija, pa je danas broj glukozinolata još uvijek varijabilan. U najnovijem istraživanju, u rodu Hesperis, identificiran je 4-O-D-apiozilglukomatronalin, čime ukupni broj glukozinolata danas broji između 89 i 136 struktura. U svrhu identificiranja izvora specifičnih glukozinolata koji bi mogli djelovati kao prekursori biološki aktivnih izotiocijanata, istraživan je sadržaj glukozinolata u biljkama. Istraživanja uključuju i razgradnju glukozinolata (enzimska, termička i kemijska) kao i biološku aktivnost (citotoksična, antimikrobna, protuupalna i dr.). Ovo djelo je dano na korištenje pod licencom Creative Commons Imenovanje 4.0 međunarodna., This review paper presents a contribution to the research of glucosinolates over the period 2005–2020, which was obtained during work at the Faculty of Chemical Technology, University of Split. After review of the comprehensive documentation, it was determined that a significant number of proposed structures lack important structural information that would be provided by NMR and MS spectroscopy, and today the number of glucosinolates is still variable. In the recent study of Hesperis genus, 4-O-D-apiosylglucomatronalin was identified, which gives the total number of glucosinolates today ranging from 89 to 136 structures. In order to identify the sources of specific glucosinolates that could act as precursors of biologically active isothiocyanates, the content of glucosinolates in plants was investigated. The study also includes the degradation of glucosinolates (enzymatic, thermal, and chemical), as well as biological activity (cytotoxic, antimicrobial, anti-inflammatory, etc.). This work is licensed under a Creative Commons Attribution 4.0 International License.

Glukozinolati, β-tioglukozidni-N-hidroksisulfati s varijabilnim bočnim lancem, su sekundarni metaboliti koji se sintetiziraju iz aminokiselina. Glukozinolati su kemijski stabilni i biološki neaktivni spojevi, a njihovom razgradnjom nastaje niz različitih biološki aktivnih razgradnih produkata. Razgrađuju se enzimskim, termičkim i kemijskim putem. U ovom radu primijenjene su različite metode izolacije uključujući hidrodestilaciju u modificiranoj aparaturi po Clevengeru, autolizu te mikrovalnu destilaciju i ekstrakciju kako bi istražili termičku, enzimsku i mikrovalovima potpomognutu razgradnju glukozinolata. Cilj rada bio je izolirati i identificirati hlapljive sumporove spojeve iz strižuše (Sisymbrium officinale (L.) Scop.). Strižuša, biljna vrsta porodice Brassicaceae, sabrana je u Splitu te na području rijeke Krke u svibnju 2019. godine. Glukozinolati su identificirani indirektno, GC-MS analizom njihovih razgradnih produkata te direktno HPLC-DAD analizom desulfatiziranih glukozinolata. U svim biljnim dijelovima strižuše (korijen, stabljika, list, cvijet), s oba područja, identificirana su tri glukozinolata glukoputranjivin (0,68-16,00 µmol/g), 4-hidroksiglukobrasicin (0,035-2,732 µmol/g) i glukobrasicin (0,244-6,397 µmol/g). U korijenu biljke još su identificirani glukosinalbin, 4-metoksiglukobrasicin i neoglukobrasicin. Hlapljivi spojevi analizirani GC-MS analizom ukazuju da je najzastupljeniji izopropil- izotiocijanat, kao razgradni produkt glukoputranjivina. Glucosinolates, β-thioglucoside-N-hydroxysulphates with variable side chains, are secondary metabolites synthesized from amino acids. Glucosinolates are chemically stable and biologically inactive compounds, and their degradation produces a number of different biologically active degradation products. They can be degraded enzymatically, thermally and chemically. Different isolation methods have been applied in this work, including hydrodistillation in the modified Clevenger apparatus, autolysis, and microwave distillation and extraction in order to investigate the thermal, enzymatic, and microwave- assisted glucosinolate degradation. The aim of the study was to isolate and identify volatile sulfur compounds from the hedge mustard (Sisymbrium officinale (L.) Scop.). Hedge mustard, a plant species of the Brassicaceae family, was collected in Split and in the Krka River area during May 2019. Glucosinolates were identified indirectly by GC-MS analysis of their degradation products and directly by HPLC-DAD analysis of desulfated glucosinolates. In all plant parts of the hedge mustard (root, stem, leaf, flower), in both areas, three glucosinolates were identified: glucoputranjivin (0.68-16.00 µmol/g), 4-hydroxyglucobrassicin (0.035-2.732 µmol/g) and glucobrassicin (0.244-6.397 µmol/g). Furthermore, glucosinalbin, 4- methoxyglucobrassicin and neoglucobrassicin were also identified at the root of the plant. The volatile compounds analyzed by GC-MS analysis indicate that the most common degradation product is isopropyl isothiocyanate, as the breakdown product of glucoputranjivin.

Glukozinolati (β-tioglukozid-N-hidroksisulfati) su skupina sekundarnih metabolita koji su zastupljeni u 16 botaničkih porodica reda Brassicales. Biološki su neaktivni, a međusobno se razlikuju po strukturi bočnog lanca. Djelovanjem enzima mirozinaze razlažu se na biološki aktivne metabolite poput izotiocijanata i nitrila. Izotiocijanati su hlapljivi sumporovi spojevi koji nastaju hidrolizom glukozinolata pri neutralnoj pH vrijednosti, a po kemijskom sastavu su esteri izotiocijanatne kiseline. Pokazuju antioksidativnu, antiupalnu, antimikrobnu, neurozaštitnu, kardiozaštitnu i antikancerogenu aktivnost. Glukozinolati i njihovi razgradni produkti odgovorni su za karakterističan miris i aromu gorušice i proizvoda od gorušice. Pri izradi rada korišteni su mikrovalni ekstrakti i destilati sjemenki divlje (Sinapis arvensis L.) i bijele gorušice (Sinapis alba L), a kemijski sastav uzoraka određen je GC-MS analizom. Analizom nisu detektirani razgradni produkti glukozinolata ni u jednom uzorku što ne znači da i nisu prisutni. Citotoksičan učinak uzoraka određen je primjenom MTT metode na staničnim linijama MBA-MB-231 (adenokarcinom dojke) i TCCSUP (karcinom prijelaznih stanica mokraćnog mjehura). Svi uzorci pokazuju citotoksičan učinak na MBA-MB-231 staničnu liniju, ali on nije nužno proporcionalan koncentraciji i trajanju inkubacije. Na staničnu liniju TCCSUP znatniji citotoksičan učinak imao je samo mikrovalni ekstrakt divlje gorušice, a u određenim slučajevima došlo je i do porasta broja stanica karcinoma. Glucosinolates (β-thioglucoside-N-hydroxysulfates) are secondary plant metabolites found in 16 botanical families of the Brassicales order. They are biologically inactive and differ by side chain structure. Myrosinase enzymes hydrolyze glucosinolates to their biologically active metabolites, such as isothiocyanates and nitriles. Isothiocyanates are volatile sulphur containing compounds which are formed by hydrolysis in pH neutral environment and are isothiocyanic acid esters by their chemical composition. They show antioxidant, anti-inflammatory, neuroprotective, cardioprotective and anticancer activity. Glucosinolates and their breakdown products are responsible for characteristic odor and aroma of mustard and mustard products. Microwave extracts and distillates of wild (Sinapis arvensis L.) and white (Sinapis alba L.) mustard seeds were used in the preparation of the paper and their chemical composition was determined by GC-MS analysis. Glucosinolate breakdown products were not detected in any sample, but that doesn’t mean that they aren’t present. Cytotoxic activity was determined by MTT assay on MBA-MB-231 (breast adenocarcinoma) and TCCSUP (urinary bladder transitional call carcinoma) cancer cell lines. All samples express cytotoxic activity on MBA-MB-231 cell line, but not necessarily proportional with sample concentration and incubation time. Only wild mustard microwave extract has had more significant effect on TCCSUP cell line and in some cases increase in number of cancer cells can be seen.

Glukozinolati su sekundarni metaboliti koji su zastupljeni u 16 botaniĉkih porodica reda Capparales, a najznaĉajnija je porodica Brassicaceae u koju se ubrajaju dobro poznate biljke iz svakodnevne prehrane kao što su kupus, brokula, cvjetaĉa, prokulice, rotkvica, repa, gorušica i druge. Po kemijskoj strukturi to su β-tioglukozidni-N-hidroksisulfati s varijabilnim boĉnim lancem. Glukozinolati su kemijski i biološki neaktivni. Njihovom enzimskom, kemijskom ili termiĉkom razgradnjom nastaje niz biološki aktivnih produkata od kojih su najviše istaţivani izotiocijanati. U ovom radu primjenjene su razliĉite metode izolacije ukljuĉujući hidrodestilaciju, autolizu te mikrovalnu destilaciju i ekstrakciju kako bi se istraţila termiĉka, enzimska i mikrovalovima potpomognuta razgradnja glukozinolata. Posebno je zanimljiva mikrovalna ekstrakcija jer predstavlja pravi primjer uporaba zelene tehnologije. Cilj ovoga rada bio je izolirati i identificirati hlapljive sumporove spojeve iz dvoredca (Diplotaxis erucoides). Biljni materijal sakupljen je u Splitu u travnju 2019. Izolacija hlapljivih spojeva provedena je hidrodestilacijom po Clevengeru, autolizom te mikrovalnom destilacijom i ekstrakcijom bez dodatka otapala. Za identifikaciju izoliranih hlapljivih spojeva korištena je GC-MS tehnika. U listu i komuški dvoredca preko razgradnog produkta alil-izotiocijanata identificiran je sinigrin, dok je u korijenu, cvijetu i stabljici identificiran sinigrin preko alil-izotiocijanata te glukonasturcin preko 2-feniletil-izotiocijanata. HPLC-DAD tehnikom preko desulfoglukozinolata potvrĊeno je njihovo prisustvo te su navedeni spojevi i kvantizirani. Koncentracija sinigrina u listu bila je 12,0996 μmol/g, u stabljici 10,4135 μmol/g, u komuški 28,8295 μmol/g, u korijenu 2,6966 μmol/g i u cvijetu je 16,3358 μmol/g. Koncentracija glukonasturcina u stabljici bila je 0,7765 μmol/g, u korijenu 16,4398 μmol/g i u cvijetu je 0,2485 μmol/g. Glucosinolates are secondary metabolites present in 16 botanical families of the Capparales order, among which the most important is Brassicaceae family, which includes well-known plants of everyday nutrition such as cabbage, broccoli, cauliflower, broomsticks, radis, beets and others. Chemically these are β-thioglucoside-N-hydroxysulfates with a variable side chain. Glucosinolates are chemically and biologically inactive. Their enzymatic, chemical, and thermal degradation result in formation of series of biologically active products, among which the most studied ones are isothiocyanates. In our study, we used different methods including hydrodistillation, autolysis followed by extraction, and microwave assisted distillation and extraction in order to investigate thermal, enzymatic, and microwave-assisted decomposition of glucosinolates. Microwave extraction is particularly interesting because it represents a true example of green technology. The aim of this study was to isolate and identify volatile sulfur compounds from white wall rocket (Diplotaxis erucoides). Plant material was collected in Split in April 2019.The isolation of volatile compounds was performed by hydrodistillation in Clevenger type apparatus, autolysis and microwave distillation and extraction without solvent addition. GC-MS technique was used to identify the isolated volatiles. The leaf and siliquae volatile extractscontained allyl-isothiocyanate, the breakdown product ofsinigrin, while the root, flower and stem next to allyl-isothiocyanate also contained 2-phenylethyl- isothiocyanate, which originated from gluconasturtiin. Glucosinolates were also confirmed and quanified by HPLC-DAD technique. Leaf contained 12,0996 μmol/g of sinigrin, stem 10,4135 μmol/g, siliquae 28,8295 μmol/g, root 2,6966 μmol/g and flower 16,3358 μmol/g. Gluconasturtiin concentration in stem was 0,7765 μmol/g, in root 16,4398 μmol/g and in flower 0,2485 μmol/g.

Glukozinolati su sekundarni metaboliti koji se sintetiziraju iz aminokiselina. Pronađeni su u 16 porodica reda Brassicales, od kojih je najznačajnija porodica Brassicaceae (kupusnjača) u koju se ubrajaju brojne biljke svakodnevno korištene u prehrani, kao što su kupus, brokula, cvjetača i druge. Po kemijskoj strukturi glukozinolati su β- tioglukozidni-N-hidroksisulfatis varijabilnim bočnim lancem. Glukozinolati su kemijski i biološki neaktivni spojevi, a njihovom razgradnjom nastaju hlapljivi sumporovi spojevi koji imaju niz ljekovitih djelovanja. Oni se razgrađuju enzimskim, termičkim i kemijskim putem. U ovom radu primijenjene su različite metode razgradnje uključujući hidrodestilaciju u modificiranoj aparaturi po Clevengeru (100 °C, 150 min), mikrovalnu destilaciju i ekstrakciju (500 W, 98 °C, 15 - 30 min) koji su primjer termičke razgradnje te autoliza, nakon koje je uslijedila ekstrakcija s diklormetanom, koja je primjer enzimske razgradnje. Cilj rada bio je izolirati i identificirati sumporove spojeve iz hrena (Armoracia rusticana P. Gaertn., B. Mey. et Scherb.). Korijen i lišće hrena sakupljeno je u samoniklom nasadu u Konavlima (u blizini Dubrovnika, Hrvatska) u ožujku 2019. godine. Za identifikaciju spojeva korištene su GC-MS i HPLC-DAD tehnika. Analizom hlapljivih spojeva identificiran je alil-izotiocijanat, kao glavni produkt razgradnje glukozinolata koji potječe od sinigrina, zajedno s feniletil-izotiocijanatom i benzenpropanonitrilom, koji potječu od glukozinolata glukonasturcina. U listu hrena prisutan je samo sinigrin, dok je u korijenu uz sinigrin prisutan i glukonasturcin. Kasnije je potvrđeno HPLC-DAD tehnikom preko desulfatiziranih (desulfoglukozinolata) produkata. Glucosinolates are secondary metabolites that are synthesized from amino acid. They were found in 16 families of Brassicales, among the most important Brassicaceae family, which includes well-known plants of everyday nutrition such as cabbage, broccoli, cauliflower and others. Chemically these are β-thioglucoside-N-hydroxysulfates with a variable side chain. Glucosinolates are chemically and bioloyically inactice compounds, and with their degradation occur volatile sulfur compound which have a number of medicinal effects. They can be broken down by enzymatic, thermal and chemical ways. In our study we used different methods of degradation which includes hydrodistillation in a modified apparatus by Clevenger (100 °C, 150 min), microwave assisted distillation and extraction (500 W, 98 °C, 15 - 30 min), which are an example of thermal degradation, and autolysis followed by extraction with dichloromethane which is an example of enzymatic degradation. The aim of this study was to isolate and identify sulfur compounds from the horseradish (Armoracia rusticana P. Gaertn., B. Mey. et Scherb.). The roots and the leaves of horseradish were collected from the wild-growing population in Konavle (near Dubrovnik, Croatia) in March 2019. GC-MS and HPLC-DAD techniques were used to identify compounds. Analysis of the volatile compounds hasidentified the presence of allyl isothiocyanate, as the main glucosinolate degradation product originating form sinigrin, along with phenylethyl isothiocyanate and benzenpropanonitrile, originating from glucosinolate gluconasturtiin. Only sinigrin was present in the leaf of horseradish, while at the root, along with sinigrin, the presence of gluconasturtiin was also confirmed. It was later confirmed with HPLC-DAD technique acros desulfated (desulfoglucosinolates) products.

Povrće iz porodice kupusnjača (Brassicaceae) karakterizira prisutnost glukozinolata, skupine sekundarnih biljnih metabolita, čiji razgradni produkti pokazuju izrazito dobru biološku aktivnost. Cilj ovo rada bila izolacija hlapljivih sastojaka iz brokule (Calabrese broccoli) i kineske brokule (Romanesco broccoli). Frakcije su izolirane iz svježeg biljnog materijala korištenjem uređaja za mikrovalnu ekstrakciju (400 W, 98°C, 35 min), a analizirane primjenom tehnike plinske kromatografije s masenom spektrometrijom (GC- MS). Frakcijama je testirana antioksidacijska aktivnost korištenjem različitih metoda: sposobnost hvatanja molekula slobodnog radikala DPPH (2,2-difenil-1-pikrilhidrazil), Briggs-Rauscher oscilacijskom metodom i metodom određivanja redukcijske aktivnosti FRAP (engl. Ferric Reducing Antioxidant Power). GC-MS tehnika je pokazala da je dominantan sumporov spoj koji je produkt degradacije u svim izolatima bio dimetil trisulfid (nastaje raspadom S-metilcistein sulfoksida). Najveća koncentracija ovog spoja određena je u frakciji brokule (0,74%) koja je ujedno pokazala i najbolju antioksidacijsku aktivnost primjenom svih korištenih metoda. The vegetable from the Brassicaceae plant family is characterized by the presence of glucosinolates, a group of secondary plant metabolites, whose degradation products show significant biological activity. The aim of this study was the isolation of the volatiles from two Brassica oleracea species: broccoli (Calabrese broccoli) and broccoflower (Romanesco broccoli). The flavour and fragrance fractions were collected from fresh plant material using microwave-assisted extraction (400 W, 98°C, 35 min) and analysed by gas chromatography-mass spectrometry (GC-MS). The fractions were tested for antioxidant activity by multiple method approach using free DPPH (2,2-diphenyl-1-picrilhydrazil) scavenging activity, Briggs-Rauscher oscillating reaction and reducing activity determination by FRAP assay (Ferric Reducing Antioxidant Power). The dominant sulfur-containing degradation product in all isolates was dimethyl trisulfide (from degradation of S-methylcysteine sulfoxide). This compound was detected in flavour fraction of broccoli (0.74%) in highest concentration, and that fraction also provided the best antioxidant activity using all applied methods.

Hlapljivi sumporovi spojevi čine veliku grupu različitih vrsta koje su pronađene u čak 43 biljne porodice i 173 biljna roda, među kojima je najvažnija porodica Brassicaceae. Glukozinolati, β-tioglukozidni-N-hidroksisulfati s varijabilnim bočnim lancem su sekundarni metaboliti identificirani u preko 16 različitih biljnih porodica. Ovim spojevima posebno obiluju biljke porodice Brassicaceae među kojima se ističe rod Brassica. Glukozinolati su kemijski i biološki neaktivni, dok su komponente koje nastaju razgradnjom istih poznate po brojnim antikancerogenim, antimikrobnim i antioksidacijskim djelovanjima. Izolacija razgradnih produkata glukozinolata vrši se destilacijom i ekstrakcijom. Identifikacija glukozinolata se provodi indirektnom metodom, kojom se GC-MS analizom utvrđuje prisustvo produkata njihove razgradnje djelovanjem enzima mirozinaze. U ovom radu obrađene su dvije vrste spomenutog roda, Brassica rupestris Raf. ssp. gongyloides L.(koraba) i Bunias erucago L. (čunjasti repušac). Rezultati analize Brassica rupestris Raf. ssp. gongyloides L. pokazali su prisutnost sinigrina, glukotropeolina, glukolimnantina, glukoerucina, glukonasturcina, ali i S-metil- L -cistein sulfoksida karakterističnog za Allium vrste. Uzorci Bunias erucago L. su prikupljeni s dva lokaliteta; Marjan u Splitu i otok Brač (Škrip). Analizom Bunias erucago L. s otoka Brača ustanovljena je prisutnost samo dvije vrste glukozinolata: glukotropeolina i glukodehidroerucina dok je u biljnom materijalu Bunias erucago L. s Marjana prisutno pet vrsta glukozinolata: glukonapin, glukotropeolin, heptil glukozinolat, glukodehidroerucin te glukoerucin. Volatile sulphur compounds make up a large group of different species found in a 43 plant families and 173 plant genera, including the most important family, Brassicaceae. Glucosinolates, β-thioglucoside-N- hydroxysulfates with variable side chains are secondary metabolites identified in over 16 different plant families. These compounds are found particularly in the Brassicaceae family, among which Brassica stands out. Isolation of the degraded glucosinolate products is performed by distillation and extraction. The glucosinolate identification is carried out by an indirect method by which is determinated the presence of degradation products by the action of the myosinase enzyme using GC-MS method. In this paper two different species of Brassica genus are processed, Brassica rupestris Raf. ssp. gongyloides L. (kohlrabi) and Bunias erucago L. (crested warty cabbage). Glucosinolates are chemically and biologically inactive, while components that are formed by their degradation are known by numerous anticancer, antimicrobial and antioxidant activities. Analysis results of Brassica rupestris Raf. ssp. gongyloides L. showed the presence of sinigrin, glucotropaeolin, glucolimnantin, glucoerucin, gluconasturitiin, but also S-methyl- L -cysteine sulfoxide characteristic for the Allium species. Samples of Bunias erucago L. are collected from two different locations; Marjan in Split and island Brač (Škrip). Analysis results of Bunias erucago L. from island Brač had shown the presence of glucotropaeoline and glucodehydroerucin while in the same plant taken from Marjan had been identified five types of glucosinolates: gluconapin, glucotropaeolin, heptyl glucosinolate, glucodehydroerucin and glucoerucin.

Hlapljivi sumporovi spojevi su metabolički produkti izrazite biološke aktivnosti, koji se mogu pronaći u biljkama, životinjama i mikroorganizmima, a posebnu raznolikost pokazuju oni koji se nalaze u biljkama. Neki od hlapljivih sumporovih spojeva su ujedno i razgradni produkti glukozinolata. Glukozinolati su biološki neaktivni i nehlapljivi spojevi, koji se mogu pronaći u 16 porodica reda Brassicales. Njihovom termičkom, enzimskom ili kemijskom razgradnjom nastaju hlapljivi spojevi poznati po svojoj biološkoj aktivnosti. U ovom radu izolirani su i analizirani hlapljivi sumporovi spojevi iz odabranih dijelova četiri biljke roda Lepidium (porodica Brassicaceae): L. graminifolium, L. latifolium, L. meyenii i L. sativum, te konjske rotkvice (Moringa oleifera) iz porodice Moringaceae. Određen je i sastav glukozinolata ovih biljaka indirektnom metodom preko njihovih razgradnih produkata. Hlapljivi izolati dobiveni hidrodestilacijom u modificiranoj aparaturi po Clevengeru i ekstrakcijom diklormetanom, analizirani su plinskom kromatografijom spregnutom s spektrometrom masa (GC-MS) i infracrvenom spektroskopijom s Fourirerovom transformacijom (FTIR). Analizom je utvrđeno da je najzastupljeniji glukozinolat u biljkama roda Lepidium glukotropeolin, koji je pronađen u sve četiri biljke. Osim glukotropeolina, identificirani su još i glukolimnantin, glukonasturcin, sinigrin, glukokohlearin i fenil glukozinolat. Od hlapljivih sumporovih spojeva koji nisu glukozinolatno vezani, pronađen je diizopropil disulfid u destilatu L. sativum. GC-MS analizom destilata i ekstrakta biljke M. oleifera nisu pronađeni razgradni produkti glukozinolata ni ostali hlapljivi sumporovi spojevi, ali je FTIR analiza pokazala prisustvo neidentificiranih spojeva. Volatile sulphur compounds are metabolites with particular biological activity found in plants, animals, and microorganisms, but the most varieties of these compounds have been found in plants. Some of the volatile sulphur compounds are also degradation products of glucosinolates. Glucosinolates are biologically inactive and non-volatile compounds found in 16 families of the Brassicales order. Volatile compounds known for their biological activity are formed by thermal, enzymatic or chemical degradation of glucosinolates. Volatile sulphur compunds from selected parts of four Lepidium spp. (Brassicaceae family) plants: L. graminifolium, L. latifolium, L. meyenii and L. sativum; and horseradish tree (Moringa oleifera) of the Moringaceae family were isolated and analysed. Glucosinolate compositions of these plants were determined by an indirect method using their degradation products. The volatile isolates collected by hydrodistillation in Clevenger-type apparatus, and extraction with dichlormethane after the hydrolysis with exogenous enzyme myrosinase were analysed with gas chromatography coupled to mass spectrometry (GC-MS) and Fourier transform infrared spectroscopy (FTIR). Analysis showed that the most widespread glucosinolate in the Lepidium spp. plants was glucotropaeolin, which had been found in all four plants. Besides glucotropaeolin, glucolimnanthin, gluconasturtiin, sinigrin, glucocohlearin and n-phenyl glucosinolate were identified. Regarding the free volatile sulphur compounds, only one was identified: diisopropyl disulphide in L. sativum. Neither glucosinolates nor free volatile sulphur compounds were found using GC-MS analysis of the distillate and the extract of the M. oleifera plant, but FTIR analysis showed a presence of unidentified compounds.

Brokula (Brassica oleracea var. italica L.) je povrće koje se smatra izvornim oblikom cvjetače, a uzgaja se zbog zelenih cvjetnih izdanaka smještenih na razgranatoj mekanoj stabljici. Brokula je bogata bioaktivnim spojevima, glukozinolatima i polifenolima te se uvelike istražuje radi njihova korisnog djelovanja na ljudsko zdravlje. U literaturi je mnogo više radova o sastavu i udjelima bioaktivnih spojeva u cvatu nego u listovima i stabljici biljke. U ovom radu analizirani su ukupni i pojedinačni glukozinolati, ukupni polifenolni spojevi te njihov antioksidacijski kapacitet u svim jestivim dijelovima brokule (cvat, listovi, stabiljka). Utvrđeno je da je količina glukozinolata najveća u listu brokule pri čemu udio indolnih prevladava nad alifatskim spojevima što vrijedi i za cvat. U stabljici je veći udio alifatskih spojeva. Udio polifenolnih spojeva u uzorcima u rasponu je od 9,70-16,11 mg GE g-1 s. tv. u stabljici, od 17,65-31,08 mg GE g-1 s. tv. u cvatu i od 22,27-34,02 mg GE g-1 s.tv. u listu brokule. Veću antioksidacijsku aktivnost pokazuju uzorci s većom koncentracijom polifenola. Ti rezultati upućuju da ne samo cvat brokule već i ostali dijelovi biljke mogu biti vrijedan izvor bioaktivnih spojeva te ih se preporuča također koristiti u prehrani. Nadalje, nusproizvodi nakon berbe, stabiljke i listovi mogli bi se koristiti kao sirovina za njihovu izolaciju te eventualnu uporabu kao dodataka prehrani., Broccoli is rich in bioactive compounds, polyphenols and glucosinolates, and as such considered as functional food and great experimental material. Previous scientific attention had mostly been focussed on composition and amounts of phytochemicals in florets rather than leaves and stem. In this research, content of total and individual glucosinolates, total polyphenols and antioxidative capacity were studied in broccoli florets, leaves and stems. Leaves showed to be the most abundant in glucosinolates with indolyl predominating aliphatic as was also the case in florets. In contrast, stem contained more aliphatic than indolyl glucosinolates. Polyphenol quantity ranged from 9.70-16.11 mg GE g-1 DW in stems, from 17.65-31.08 mg GE g-1 DW in florets and from 22.27-34.02 mg GE g-1 DW in leaves and was the highest in leaves irrespective of extraction method. It was determined that samples containing the highest amounts of polyphenols also had the highest antioxidant activity.

Glukozinolati su složena klasa spojeva tioglukozidne strukture svojstvena biljkama porodice Cruciferae, u koju ubrajamo značajne kulture kao što su uljana repica, kupus, cvjetača, brokula, koraba, kelj, hren, gorušica. Sve biljke koje sadrže glukozinolate sadrže i endogeni enzim hidrolize - mirozinazu (β-tioglukozid-glukohidrolaza, EC 3.2.1.1) u odijeljenim dijelovima biljke. Oštećenjem tkiva oni dolaze u kontakt s enzimom mirozinazom te ovisno o strukturi glukozinolata nastaju različiti biološki aktivni produkti razgradnje, koji imaju čitav niz bioloških svojstava, od toksičnih do antikarcinogenih. Udio i sastav glukozinolata razlikuje se između biljnih vrsta te izmedu sorata istih vrsta te se mogu razlikovati unutar biljne jedinke ovisno o stupnju razvoja i vrsti organa. Različiti uvjeti okoliša u kojima se biljka nalazi, kao što su svjetlo, nutritivni status biljke, mehaničko oštećenje, infekcija fitopatogenima i oštećenja uzrokovana napadom insekata, mogu uzrokovati značajne promjene u sastavu i udjelu glukozinolata. Stoga postoji niz teorija o potencijalonoj ulozi glukozinolata u biljci, iako se njihova uloga u obrani biljke od insekata i fitopatogena smatra primarnom., Glucosinolates are sulfur- and nitrogen-containing plant secondary metabolites common in the Brassicaceae and related plant families. In the plant, they coexist with an endogenous β-thioglucosidase (EC 3.2.3.1) called myrosinase and upon plant tissue disruption, glucosinolates are released at the damage site and become hydrolyzed by myrosinase. The chemical nature of the hydrolysis products depends on the structure of the glucosinolate side chain, plant species and reaction conditions. Glucosinolate pattern differs between species and ecotype as well as between and even within individual plants, depending on developmental stage, tissue and photoperiod. A number of environmental conditions such as light plant, nutritional status, fungal infection, wounding and insect damage can alter the glucosinolate pattern significantly. The change of the glucosinolate profile by several environmental factors has brought forward different theories regarding their potential roles in the plant. However, the most accepted theory is that the glucosinolate-myrosinase system is involved in defense against herbivores and pathogens.

Glukozinolati su sekundarni biljni metaboliti ß-tioglukozidne strukture koji hrani biljnog podrijetla daju aromu, miris i okus. Smatraju se fitokemikalijama što znači da mogu predstavljati prevenciju ljudskom organizmu od različitih bolesti, a naročito kod pojave karcinoma. Glavni izvor glukozinolata u ljudskoj prehrani su biljke iz porodice Brassicaceae u koju pripadaju kupus, brokula, cvjetača, repa, kelj, hren. Glukozinolati su fiziološki neaktivni i kemijski stabilni i tek njihovom razgradnjom djelovanjem enzima mirozinaze dolazi do oslobađanja raznovrsnih hlapljivih spojeva (izotiocijanata, tiocijanata, nitrila). U radu će biti opisana struktura, biodostupnost glukozinolata kao i njihov sadržaj u biljkama, njihovo oslobađanje enzimskom ili neenzimskom razgradnjom, stabilnost tijekom procesiranja te pozitivno djelovanje na ljudsko zdravlje. Osim što njihov unos ima pozitivan učinak na ljudski organizam, glukozinolati i njihovi razgradni spojevi mogu imati i antinutritivno djelovanje. Glucosinolates are plant secondary metabolites of ß-thioglucoside structure that are responsible for flavor, aroma and taste of foods of plant origin. They are considered phytochemicals which means that they can protect the human body from various diseases, especially with the onset of cancer. The main sources of glucosinolates in the human diet are plants from the family Brassicaceae, like cabbage, broccoli, cauliflower, beets, savoy cabbage, horseradish. Glucosinolates are physiologically inert and chemically stable. Activation of the enzyme myrosinase leads to the formation of various volatile compounds (isothiocyanates, thiocyanate, nitrile). This paper deals with structure, bioavailability of glucosinolates and their content in plants, their release by enzymatic or non-enzymatic degradation, stability during processing and positive effect on human health. Next to positive effect on the human body, glucosinolates and their degradation compounds may have antinutritive activity.

Danas ljudi sve više konzumiraju hranu računajući i na njene sastojke koji se nalaze u malim količinama, a koji mogu imati povoljan utjecaj na zdravlje. Takve sastojke nazivamo biološki aktivne komponente hrane. Kvaliteta ovih supstanci ne mjeri se njihovom količinskom zastupljenosti, nego razinom sposobnosti da tokom svoje biološke aktivnosti mogu neutralizirati utjecaje štetnih tvari u ljudskom organizmu. Sve biološki aktivne komponente se međusobno razlikuju kako po kemijskoj strukturi tako i po funkciji u organizmu. U neke od ovih sastojaka ubrajaju se vitamini i minerali, karotenoidi, fenolni spojevi, flavonoidi, fitosteroli, izoflavoni, enzimi, tanini, glikozidi, alkaloidi, i druge. Glukozinolati pripadaju takvim tvarima koje su intenzivno istraživane na području prehrane i medicine, te se smatra da imaju antikancerogena svojstva i sudjeluju u prevenciji različitih bolesti. Zeljasto povrće je izvor dijetalnih glukozinolata, posebice jedne velike skupine – sumporni glukozinolati. Ovi spojevi ostaju netaknuti, osim ukoliko nisu u kontaktu sa enzimom mirozinazom, do kojega dolaze povredom tkiva od strane štetnika ili žvakanjem istog. Mirozinaza otpušta glukozu i izdvaja izotiocijanate iz povrća. Izotiocijanati su spojevi sa kemijskom grupom -N = C = S, koja je formirana zamjenom sumpora za kisik u izocijanat grupi. Povrće sadrži razne glukozinolate, od kojih svaki hidrolizom tvori drugačiji izotiocijanat. Izotiocijanati poput sulforafana pomažu u prevenciji raka eliminiranjem mogućih kancerogenih tvari iz tijela. Epidemiološke studije su dokazale da izloženost ljudi iztiocijanatima, konzumacijom zeljastog povrća, smanjuje rizik pojave raka, te imaju istovremeno i zaštitnu ulogu. Međutim, iako su prisutni u velikim količinama, vrlo su osjetljivi spojevi, te kuhanjem, rezanjem i obradom mikrovalovima smanjuje se njihova bioraspoloživost. Today, people are increasingly consuming food counting on its ingredients that are found in small quantities, which can have a positive impact on health. Such substances are called biologically active food components. The quality of these substances is not measured by the quantitative representation, but the level of skills that during their biological activity can counteract the effects of harmful substances in the human body. All biologically active components are different in chemical structure and their function in the body. Some of these ingredients include vitamins, minerals, carotenoids, phenolic compounds, flavonoids, phytosterols, isoflavones, enzymes, tannins, glycosides, alkaloids, and others. Glucosinolates belong to such substances, which have been extensively studied in the field of nutrition and medicine, and is considered to have anti-cancer properties and participate in the prevention of various diseases. Cruciferous vegetables are a source of dietary glucosinolates, especially a large group - sulfur glucosinolates. These compounds remain intact, unless they are in contact with the enzyme myrosinase, which occurs tissue injury by pests or chewing it. Myrosinase releases glucose and isothiocyanate extracts from vegetables. Isothiocyanates are compounds with the chemical group-N = C = S, which is formed by substituting sulfur for oxygen in the isocyanate group. Vegetables contain a variety of glucosinolates, each form a different isothiocyanate by hydrolysis. Isothiocyanates, such as sulforaphane, help prevent cancer by eliminating potential carcinogens from the body. Epidemiological studies have shown that exposure to isthiocyanates, consumption of green leafy vegetables, reduces cancer risk, and have both a protective role. However, although present in large quantities, they are very sensitive compounds, so cooking, cutting and microwaves reduces their bioavailability.