Ivica Ljubenkov

Phenolic Profile, Antioxidant Capacity, and Antimicrobial Activity of Leaf Extracts from Six Vitis vinifera L. Varieties

The phenolic composition, antioxidant activity, and antimicrobial activity of extracts from vine leaves of six grape varieties collected in May, August, and September was studied. The phenolic potential of the extracts was dependent on variety and picking-time. Extracts of leaves collected in September were the richest in total phenols, flavonoids, flavonols, and stilbenes. The antioxidant properties determined by ferric reducing antioxidant power and 2,2-diphenyl-1-picrylhydrazyl assays, and antimicrobial activity against Stapylococcus aureus, Bacillus cereus, Campylobacter jejuni, Escherichia coli, and Salmonella Infantis were good and in correlation with the chemical composition changes of the leaf extracts. The results indicated that leaves remaining on the vine in September after the grape harvest could be especially promising as an inexpensive source of effective antioxidant/antimicrobial agents.

Seasonal Variations of Phenolic Compounds and Biological Properties in Sage (Salvia officinalis L.)

The aim was to investigate the phenolic content, antioxidant capacity, and antibacterial activity of Dalmatian sage (Salvia officinalis L.) leaves collected during different vegetation periods. Separation and quantification of the individual phenols were performed by reversed‐phase (RP)‐HPLC coupled with a PDA (photodiode array) detector and using an internal standard, while the contents of total phenols, flavonoids, flavones, and flavonols were determined spectrophotometrically. The antioxidant properties of the sage leaf extracts were evaluated using five different antioxidant assays (FRAP, DPPH, ABTS, BriggsRauscher reaction, and β‐carotene bleaching). The antimicrobial activity of the extracts was tested against two Gram‐positive (Bacillus cereus and Staphylococcus aureus) and two Gram‐negative (Salmonella Infantis and Escherichia coli) bacterial reference strains. All extracts were extremely rich in phenolic compounds, and provided good antioxidant and antibacterial properties, but the phenophase in which the leaves were collected affected the phenolic composition of the sage extracts and consequently their biological activity. The May Extract, the richest in total flavonoids, showed the best antioxidant properties and the highest antimicrobial activity. Thus, collection of the plants during May seems the best choice for further use of them in the pharmaceutical and food industry.

Antioxidant and vasodilatory effects of blackberry and grape wines.

In contrast to the well-described various biological effects of grape wines, the potential effects of commonly consumed blackberry wine have not been studied. We examined in vitro antioxidant and vasodilatory effects of four blackberry wines and compared them with the effects of two red and two white grape wines. Although some blackberry wines had lower total phenolic content relative to the red grape wines, their antioxidant capacity was stronger, which may be related to a higher content of non-flavonoid compounds (most notably gallic acid) in blackberry wines. Although maximal vasodilation induced by blackberry wines was generally similar to that of red wines, blackberry wines were less potent vasodilators. Vasodilatory activity of all wines, in addition to their flavonoid and total phenolic content, was most significantly associated with their content of anthocyanins. No association of vasodilation with any individual polyphenolic compound was found. Our results indicate the biological potential of blackberry wines, which deserves deeper scientific attention.

Influence of the phenophase on the phenolic profile and antioxidant properties of Dalmatian sage.

This study aimed to research the influence of phenophase on the phenolic profile (phenolic acids, flavonoids and stilbenes) and related antioxidant properties of sage, one of the most characteristic plants of Dalmatian karst. The total phenol and flavonoid contents in sage leaves were determined spectrophotometrically, while the principal phenolics were determined using HPLC-RP-DAD. Antioxidant properties of this well known medicinal plant were determined as free radical scavenging activity (DPPH), ferric reducing/antioxidant power (FRAP) and by Briggs-Rauscher oscillating reaction (BR). The results strongly indicate that Dalmatian sage leaves are rich source of valuable phenolics, mainly phenolic acids, with extremely good antioxidant properties. The presence of resveratrol or its derivatives was confirmed in all extracts. The best results for total phenols and flavonoids, as well as the best antioxidant properties were obtained for May sage, while the highest amounts of catechin monomers and compounds from group of stilbenes were found in February extract.

Chemical Composition and Biological Activity of Allium cepa L. and Allium × cornutum (Clementi ex Visiani 1842) Methanolic Extracts

Here, we report a comparative study of the phytochemical profile and the biological activity of two onion extracts, namely Allium cepa L. and Allium × cornutum (Clementi ex Visiani 1842), members of the family Amaryllidaceae. The identification of flavonoids and anthocyanins, and their individual quantities, was determined by high-performance liquid chromatography (HPLC). The potency of both extracts to scavenge free radicals was determined by the DPPH (2,2′-diphenyl-1-picrylhydrazyl) radical-scavenging activity and oxygen radical absorbance capacity (ORAC) methods. The DNA protective role was further tested by the single-cell gel electrophoresis (COMET) assay and by Fenton’s reagent causing double-strand breaks on the closed circular high copy pUC19 plasmid isolated from Escherichia coli. In the presence of both extracts, a significant decrease in DNA damage was observed, which indicates a protective role of Allium cepa and Allium × cornutum on DNA strand breaks. Additionally, cytotoxicity was tested on glioblastoma and breast cancer cell lines. The results showed that both extracts had antiproliferative effects, but the most prominent decrease in cellular growth was observed in glioblastoma cells.

Evaluation of Olive Fruit Lipoxygenase Extraction Protocols on 9- and 13-Z,E-HPODE Formation

In plant tissues, enzymes implicated in the lipoxygenase (LOX) pathway are responsible for the hydroperoxydation of polyunsaturated fatty acids, ultimately leading to the production of small chemical species involved in several physiological processes. During industrial olive oil production, these enzymes are activated upon crushing and grinding of olive fruit tissue, subsequently leading to the synthesis of volatile compounds responsible for the positive aroma and flavor of the oil. An investigation of LOX activity during olive fruit ripening and malaxation could assist in the production of oils with favorable aroma and taste. Therefore, a reliable method for olive LOX purification is crucial. Here we report a critical review of six LOX extraction protocols, two of which have shown minimum enzyme activity, possibly leading to misconceptions in the interpretation of experimental data. Future research concerning olive LOX should employ extraction methods that preserve enzyme activity.

The Content of Biogenic Amines in Croatian Wines of Different Geographical Origins

Samples of white and red wines produced in two different wine-growing regions, coastal (Dalmatia) and continental (Hrvatsko zagorje) of Croatia, were analysed for biogenic amines content. Biogenic amines content was determined, and its concentration levels were associated with the geographical origin of the wine. Due to its high sensitivity, HPLC method with ultraviolet detector was used, including the derivatisation step with dansyl chloride. The method was applied to detect and quantify 11 biogenic amines in 48 red and white wines. It was found that both Dalmatian red and white wines are characterised by tryptamine (0.23–1.22 mg L−1), putrescine (0.41–7.5 mg L−1) and ethanolamine (2.87–24.32 mg L−1). White wines from the Hrvatsko zagorje region are characterised by content of isopentylamine (0.31–1.47 mg L−1), putrescine (0.27–1.49 mg L−1) and ethanolamine (3.80–17.96 mg L−1). In contrast to white wines from the Hrvatsko zagorje region, in the red wines, all biogenic amines except ethylamine, were found and equally presented.

Bemisia tabaci MED Population Density as Affected by Rootstock-Modified Leaf Anatomy and Amino Acid Profiles in Hydroponically Grown Tomato

Bemisia tabaci is one of the most devastating pests in tomato greenhouse production. Insecticide resistance management for B. tabaci requires a novel approach that maximizes non-chemical methods for pest control. The aim of this study was to test the effects of rootstocks on B. tabaci populations in hydroponically grown tomato plants. In order to contribute to the better understanding of the mechanisms defining the attractiveness of plant to the aerial pest, the effects of rootstocks on leaf anatomy and the amino acid composition of phloem sap were assessed. A two-factorial experimental design was adopted using cultivars (rootstock cultivars and Clarabella) grown as either non-grafted or grafted with cultivar Clarabella as a scion. The rootstock cultivars included Arnold, Buffon, Emperador, and Maxifort. A reduction in B. tabaci density was observed using all rootstock cultivars. The number of adult individuals per leaf was 2.7–5.4 times lower on rootstock cultivars than on Clarabella. The number of large nymphs per square centimeter was at least 24% higher on non–grafted Clarabella compared with all other treatments. The leaf lamina thickness and mesophyll thickness were lower in self-grafted Clarabella than in non-grafted or in one grafted on rootstock cultivars; however, the extent of this reduction depended on the rootstock. The leaves with thinner laminae were generally less attractive to B. tabaci. Eighteen amino acids were detected in the exudates of phloem sap. In all treatments, the most abundant amino acid was γ-aminobutyric acid (GABA), followed by proline, serine, alanine, and histidine. The scion cultivar Clarabella was the most attractive to B. tabaci and had a higher content of leucine than did rootstock cultivars, and a higher content of lysine compared to Buffon and Maxifort. The features modified by rootstock such are changes in leaf anatomy can affect the attractiveness of plants to B. tabaci. Thus, the grafting of tomato could constitute a valuable tool in an integrated management strategy against this aerial pest.