Primjena kultura stanica za određivanje biološke aktivnosti spojeva iz biljaka

Prirodni proizvodi poput žitarica, orašastih plodova, voća, povrća i ljekovitog bilja sadrže brojne biološki aktivne tvari (fenolne spojeve, glukozinolate, karotenoide, alkaloide i dr.) koje mogu imati različite povoljne, ali i nepovoljne učinke na organizam. Povoljni učinci na zdravlje ljudi povezuju se s antitumorskim, antialergijskim, antibiotičkim, antioksidacijskim i drugim svojstvima koje mogu posjedovati biološki aktivne tvari iz biljaka. Utvrđeno je da djelovanje navedenih tvari ima zaštitni učinak kod širokog raspona tumorskih oboljenja, smanjuje mogućnost pojave koronarnih bolesti, srčanog udara te štiti organizam od oksidativnog stresa uzrokovanog slobodnim radikalima. Stoga se sve više ispituje primjena biološki aktivnih tvari iz biljaka kao izvora novih lijekova, pri čemu se u istraživanjima često koriste in vitro testovi koji uključuju uporabu različitih humanih staničnih linija. Stanične linije koriste se u procjeni toksičnosti kemikalija i uzoraka iz okoliša te predstavljaju vrlo koristan alternativni pristup u (eko)toksikološkim istraživanjima. Također, često se koriste pri ispitivanju biološke aktivnosti spojeva prisutnih u biljkama, osobito voću i povrću, te raznih proizvoda koji se baziraju na tim sirovinama. Interakcija ispitivane tvari i bioloških sustava odvija se ponaprije na staničnoj razini, na čemu se temelji primjena in vitro staničnih kultura, koje omogućuju brže i ekonomičnije studije u odnosu na in vivo istraživanja. Kod in vitro testova najčešće se određuje tzv. bazalna citotoksičnost, kojom se određuje učinak ispitivane tvari na preživljenje stanica u kulturi, a tako dobiveni rezultati mogu poslužiti kao smjernice za daljnja in vivo istraživanja

Glucosinolates potential role in plant defence

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

Mehanizmi toksičnosti ionskih kapljevina

Over the past three decades a growing awareness of environmental protection prompted the development of so-called green and sustainable technologies. Therefore, academic and wider community intensively explores new chemicals and safer, more energy efficient processes based on a rational compromise between economic, social, and environmental requirements. Due to low volatility and stability, ionic liquids emerged as a potential replacement for traditional volatile and harmful organic solvents. Various studies have been carried out to validate the green character of ionic liquids, whereby data published suggest that these compounds, due to their relatively high toxicity and poor biodegradability, could have an extremely negative impact on the environment. This paper presents the current knowledge on the toxicity of ionic liquids, with a special emphasis on the mechanisms by which this group of compounds causes changes in the morphology and physiology of organisms at different organisational levels of the ecosystem.U protekla tri desetljeća sve više raste svijest ljudi o potrebi zaštite okoliša, pa se velika pozornost pridaje tzv. zelenim i održivim tehnologijama. Stoga se u akademskim sredinama, a i u široj zajednici, intenzivno istražuju nove kemikalije te sigurniji i energetski učinkovitiji procesi koji se zasnivaju na prihvatljivom kompromisu između ekonomskih, socijalnih i ekoloških zahtjeva. Ionske se kapljevine zbog neznatne hlapljivosti i stabilnosti smatraju potencijalnom zelenom zamjenom za tradicionalna i škodljiva organska otapala. Kako bi se potvrdio zeleni karakter ionskih kapljevina, posljednjih godina provode se različita istraživanja njihova učinka na okoliš. Do sada objavljeni podaci upućuju na to da bi ova skupina spojeva, zbog relativno visoke toksičnosti i slabe biorazgradljivosti, ipak mogla imati izrazito negativan utjecaj na okoliš. U ovom radu prikazana su dosadašnja saznanja o toksičnosti ionskih kapljevina, s naglaskom na mehanizme kojima ova skupina spojeva uzrokuje promjene u morfologiji i fiziologiji organizama koji se nalaze na različitim organizacijskim razinama ekosustava

Bioactive compounds glucosinolates and polyphenols in florets, leaves and stems of broccoli (Brassica oleracea var. italica L.)

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

Extraction of Proanthocyanidins and Anthocyanins from Grape Skin by Using Ionic Liquids

U ovom je radu ispitana mogućnost uporabe osam različitih imidazolijevih ionskih kapljevina kao novih otapala za ekstrakciju flavonoida iz pokožice grožđa, te su rezultati uspoređeni s konvencionalnom ekstrakcijom pomoću organskog otapala. Struktura aniona, kationa i koncentracije ionskih kapljevina značajno su utjecali na iskorištenje ekstrakcije. Najveća koncentracija proantocijanidina dobivena je pomoću 2,5 mol/L 1-butil-3- -metilimidazolijevog bromida [C4mim][Br], a antocijana pomoću 2,5 mol/L 1-etil-3- -metilimidazolijeva bromida [C2mim]. Statistički bitno veće koncentracije ukupnih i svih slobodnih antocijana dobivene su ekstrakcijom pomoću 2,5 mol/L [C2mim][Br] i 2,5 mol/L 1- -metilimidazolijevog hidrogen sulfata [HM][HSO4], u usporedbi s konvencionalnim otapalom. Nadalje, 2,5 mol/L [C4mim][Br] i 2.5 mol/L 1-(4-sulfobutil)-3-metilimidazolijevog hidrogen sulfata [S4mim][HSO4] imali su znatno veću selektivnost prema antocijanin-3-O- -acetilmonoglukozidu i antocijanin-3-(6-O-p-kumaril)monoglikozidu.In this study, eight different types of imidazolium-based ionic liquids (ILs) were applied as new solvents in the extraction of flavonoids from grape skin, and compared to the conventional organic solvent extraction that was not reported earlier. The structure of anions, cations and concentration of ILs significantly affected extraction yields. The highest mass fractions of proanthocyanidins and anthocyanins were obtained with 2.5 mol/L of 1-butyl-3-methylimidazolium bromide [C4mim][Br] and 2.5 mol/L of 1-ethyl-3-methylimidazolium bromide [C2mim][Br], respectively. The studied ILs provided an excellent preliminary result in the extraction of anthocyanins. Significantly higher mass fractions of total and all free anthocyanins were extracted with 2.5 mol/L of [C2mim][Br] and 2.5 mol/L of 1-methylimidazolium hydrogen sulfate [mim][HSO4] than with conventional solvent with the exception of anthocyanin-3-O-acetylmonoglucosides in the latter. On the other hand, 2.5 mol/L of [C4mim][Br] and 2.5 mol/L of 1-(4-sulfobutyl)-3-methylimidazolium hydrogen sulfate [sC4mim][HSO4] showed significantly higher selectivity towards anthocyanin-3-O-acetylmonoglucosides and anthocyanin-3-(6-O-p-coumaroyl)monoglucosides

The importance and benefits of using the film in the production of vegetables

U ovom radu ispitan je utjecaj kadmija na rast kelja (Brassica oleracea var. sabauda) i kupusa (Brassica oleracea var. capitata), distribucija kadmija u pojedine dijelove biljaka uzgojenih u tlu s različitim udjelima kadmija te njihov fitoekstrakcijski potencijal. Također je praćen i utjecaj povećanog udjela kadmija u tlu na sadržaj polifenola, kako bi se utvrdila moguća povezanost tih sekundarnih metabolita i odgovora biljke na stres uzrokovan prisutnošću teških metala u tlu. Dobiveni rezultati mogu biti od osobitog značaja za naše područje budući da su kupus i kelj važne i relativno često korištene namirnice u našoj prehrani te iznimno dobro uspijevaju u našem podneblju, stoga bitna je spoznaja kako one mogu akumulirati značajne količine teških metala prilikom rasta na onečišćenom tlu. Njihova dugotrajna konzumacija može nepovoljno utjecati na zdravlje ljudi i životinja.In this work the effect of cadmium on the growth of kale (Brassica oleracea var. sabauda) and cabbage (Brassica oleracea var. capitata), distribution of cadmium in certain parts of the plants grown in soil with different concentrations of cadmium and their phytoextraction potential, was investigated. As well as the impact of an increased effect of cadmium content in the soil on polyphenol content, to be able to determine the possible relationship of these secondary metabolites and plant responses to stress caused by the presence of heavy metals in soil. The obtained results could be of particular importance for our region since cabbage and kale are important and relatively frequently used food in our diet and they extremely grow well in our climate in our climate. Their long-term consummation can have adversely effect on the human and animal health

Deep Eutectic Solvents – Creating a Green Solvent for the Future through Rational Design

Niskotemperaturna eutektička otapala intenzivno se proučavaju kao ekološki prihvatljiva zamjena za štetna, u industriji tradicionalno sveprisutna organska otapala. Struktura te fizikalno-kemijska svojstva tih otapala mogu se dizajnirati za specifične namjene tako da zadovoljavaju tehnološke i ekonomske zahtjeve industrije, stoga su se pokazala vrlo učinkovitima u različitim područjima poput elektrokemije, organske sinteze i (bio)katalize, biotehnologije i prehrambene tehnologije, farmaceutskog inženjerstva te biomedicine. Ovaj rad donosi pregled znanstvene literature vezane uz karakteristična svojstva, pripravu i primjenu, ali i izazove primjene niskotemperaturnih eutektičkih otapala u industrijskom mjerilu. Ovo djelo je dano na korištenje pod licencom Creative Commons Imenovanje 4.0 međunarodna.Deep eutectic solvents arose as substitutes for harmful industrial solvents, as they fully meet the principles of green chemistry. Their structure and physicochemical properties can be designed for specific purposes, so they have proven to be effective in various fields such as electrochemistry, organic synthesis and (bio)catalysis, biotechnology and food technology, pharmaceutical engineering and biomedicine, where they can fulfil the technological and economic demands of the industry. This paper provides a literature overview of the characteristic properties, preparation, and application of deep eutectic solvents, but also the challenges of their application on an industrial scale. This work is licensed under a Creative Commons Attribution 4.0 International License

Review of deep eutectic systems from laboratory to industry, taking the application in the cosmetics industry as an example

Publisher Copyright: © 2022 The AuthorsGreen and sustainable chemistry principles have been employed in different industries to create greener and safer chemicals and processes, by replacing hazardous solvents with safer and more efficient ones. A state-of-the-art analysis on deep eutectic systems (DES) shows their consistent growth in the fields of chemistry, particularly as extraction solvents, mainly of bioactive compounds from natural sources. In this sense, a patent review was carried out, to highlight the relevance of DES, formed by natural compounds, in the cosmetic industry and the commercial added value of this technology at the industrial scale. Despite their good extraction efficiency, DES physicochemical properties need to be considered, as they may present some limitations in the process scale-up. This implies the study of several parameters, such as the cost of raw materials, the investment necessary to adapt or implement new equipment, energy consumption (for instance for pumping, heating, and stirring), and treatment of any by-products and wastes, in both the upstream and the downstream operations. Due to their inherent properties, DES can also be used as stabilizing agents of the bioactive compounds extracted, allowing their direct use in final products, namely in cosmetics, which is translated into a reduction of the downstream costs. This work further reports a case study of the extraction of grape anthocyanins with DES, and the necessary steps required to take this process from a laboratory scale to an industrial scale, targeting the cosmetic industry.publishersversionpublishe

Synthesis, application in biotransformations and cytotoxicity of selected imidazolium-based ionic liquids

Pripravljene nove imidazolijeve ionske tekućine ispitane su kao otapala u lipazom kataliziranoj sintezi izoamil-acetata te je uspostavljena visokoproduktivna kontinuirana sinteza izoamil-acetata u mikroreaktoru s nasutim slojem imobilizirane lipaze (61 mmol L−1 min−1). Primjenom nekonvencionalnih metoda u pripravi ionskih tekućina moguće je znatno skratiti vrijeme reakcije i smanjiti potrošnju energije. Ispitivanjem citotoksičnosti na CCO staničnoj liniji utvrđeno je da imidazolijeve ionske tekućine uglavnom ispoljavaju umjerenu toksičnost (EC50 = 0,1 – 10 mmol L-1), induciranu oštećenjem stanične membrane i posljedičnom nekrozom.Novel imidazolium ionic liquids were prepared and tested as solvents for enzyme-catalyzed isoamyl acetate synthesis and highly productive continuous esterification in an immobilized lipase packed-bed microreactor was developed (61 mmol L−1 min−1). The use of unconventional methods for ionic liquids’ synthesis contributes to the reduction of preparation time as well as energy consumption. Results of in vitro cytotoxicity assays indicate moderate toxicity of imidazolium ionic liquids (EC50 = 0,1 – 10 mmol L-1) through the mechanism of membrane disruption followed by necrosis