Branka Mozetič Vodopivec

Ultrasonic Extraction of Phenols from Olive Mill Wastewater: Comparison with Conventional Methods

Recovery of phenols from olive mill wastewater (OMWW) was studied, comparing five sample preparation methods: filtration, solid-phase (SPE), liquid-liquid (LLE) and ultrasonic (US)-assisted extraction of liquid and solid (freeze-dried) OMWW. Results showed that ultrasonication is a good alternative to conventional solvent extractions, providing higher recoveries at both levels of individual and total phenol yields. Sonication of liquid OMWW in organic solvent was more efficient vs its nonassisted counterpart (agitation), but did not provide a representative phenol chromatogram due to ethyl acetate use. By contrast, the US-assisted extraction of freeze-dried OMWW (3 × 20 min) in 100% methanol (1.5 g/25 mL, w/v) offered the highest qualitative-quantitative phenol yields without any US-induced alterations. Moreover, freeze-drying is an excellent preservation of initial liquid OMWW, holding a great potential for delayed analysis. This study is also the first report that Slovenian OMWW may be utilized as a valuable source of phenols, especially hydroxytyrosol and tyrosol.

Phenolic Profiling of Olives and Olive Oil Process-Derived Matrices Using UPLC-DAD-ESI-QTOF-HRMS Analysis

All of the matrices entailed in olive oil processing were screened for the presence of known and new phenol constituents in a single study, combining an ultra high pressure liquid chromatography system with diode array and electrospray ionization quadrupole time-of-flight high resolution mass spectrometry (ESI-QTOF-HRMS) detection. Their trail was followed from the fruit (peel/pulp and stone) to the paste and final products, i.e. pomace, wastewater, and oil, providing important insight into the origin, disappearance, and evolution of each during the operational steps. Eighty different phenols, composed of fruit native representatives and their technologically formed and/or released derivatives, were detected in six olive matrices and fully characterized on the basis of HRMS and UV-vis spectroscopic data. In addition to phenols already known in olive matrices, four new molecular formulas were proposed and three new tentative identities assigned to newly discovered phenols, i.e., β-methyl-OH-verbascoside, methoxynüzhenide, and methoxynüzhenide 11-methyl oleoside.

Olive fruit phenols transfer, transformation and partition trail during laboratory-scale olive oil processing

This work is the most comprehensive study on the quantitative behavior of olive fruit phenols during olive oil processing, providing insight into their transfer, transformation, and partition trail. In total, 69 phenols were quantified in 6 olive matrices from a three-phase extraction line employing ultra high pressure liquid chromatography-diode array detection analysis. Crushing had a larger effect than malaxation in terms of phenolic degradation and transformation, resulting in several new evolutions of respective derivatives. The peel and pulp together confined 95% of total fruit phenols, while stone only 5%. However, only 0.53% of all ended-up in olive oil, nearly 6% in wastewater, and 48% in pomace. Secoiridoids were the predominant class in all matrices, though represented by different individuals. Their partition behavior was rather similar to other phenolic classes, where with few minor exceptions only aglycones were partitioned to the oil, while other glycosides were lost with the wastes.

Isotopic and elemental characterisation of Slovenian apple juice according to geographical origin: Preliminary results

This study examined the applicability of stable isotope and multi-element data for determining the geographical origin of fresh apple juices. Samples included three apple cultivars (Idared, Golden Delicious and Topaz) harvested in 2011 and 2012 from five different geographical regions of Slovenia. Regional discrimination of the juice samples was most successful when using linear discriminant analysis (LDA) and taking into account the following parameters: δ(2)H and δ(18)O content of juice water; δ(15)N and δ(13)C content of the pulp, (D/H)I and (D/H)II in ethanol and the concentration of S, Cl, Fe, Cu, Zn and Sr. Overall prediction ability was 83.9%. The factors that best distinguished the different types of cultivar were the δ(2)H and δ(18)O content of fruit juice water; the δ(13)C and (D/H)I content of ethanol; and the concentration of S, Mg, K, Cu, and Ti. Prediction ability, taking into account all ten parameters, was 75.8%.

Soil and foliar fertilization affects mineral contents in Vitis vinifera L. cv. 'rebula' leaves

Grapevine nutrient oversupply as well as shortage can both result in unbalanced vine growth and poor grape production. Mineral fertilization is a powerful tool also in viticulture in order to increase yields and improve grape quality. The trial carried out in Slovenian winegrowing region investigated the effects of basic fertilization coupled with soil or foliar applications of fertilizers containing Mg and Fe on the concentration of K, Mg, Fe and Zn in the grapevines leaves at berry set and veraison. The results obtained in a 2-years study (2008-09) with seven different fertilization treatments (along with control) showed that fertilization with K decreased Mg uptake almost on a half comparing to untreated vines, resulting in basal leaves chlorosis. High K/Mg ratio, determining low Mg uptake, was not lowered in our trial neither by foliar spraying nor by fertirigation with MgSO 4 solution. On the other hand, Mg application in addition to Fe supply had same effect on soil Zn uptake and its accumulation in the leaf petioles. Moreover, foliar Fe fertilization enhanced Fe levels in blades at berry set and to a lesser extent also at veraison.

Differences in the structure of anthocyanins from the two amphibious plants, Lobelia cardinalis and Nesaea crassicaulis

The foliar anthocyanin profiles of two amphibious plants, Nesaea crassicaulis and Lobelia cardinalis were analysed for the first time. N. crassicaulis produced very simple anthocyanins, achieving the highest concentrations when grown submerged. In contrast, L. cardinalis produced leaves with a high content of very complex, acylated anthocyanins, especially when growing emergent. Anthocyanins were separated by high performance liquid chromatography. Nesaea crassicaulis anthocyanins were identified according to their fragment mass spectra and ultra-visible-violet spectral characteristics and 1D and 2D NMR spectra as -3,5-di-O-β-glucosides of delphinidin, cyanidin, petunidin, malvidin and peonidin as well as cyanidine and peonidin-3-O-β-glucoside. In L. cardinalis cyanidin-3-O-[6-O-(4-O-E-p-coumaroyl-O-α-rhamnopyranosyl)-β-glucopyrano]-5-O-β-glucopyranoside was the major anthocyanin and contributed more than 98% of total anthocyanin content. The remaining 2% was made up by cyanidin-3-O-[6-O-(4-O-E-caffeoyl-O-α-rhamnopyranosyl)-β-glucopyrano]-5-O-β-glucopyranoside and pelargonidin-3-O-[6-O-(4-O-E-p-coumaroyl-O-α-rhamnopyranosyl)-β-glucopyrano]-5-O-β-glucopyranoside.