Establishing the chromatographic fingerprints of flavan-3-ols and proanthocyanidins from rose hip (Rosa sp.) species.

The profile of flavan-3-ols and proanthocyanidins in five different Rosa species (R. canina, R. glutinosa, R. rubiginosa, R. multiflora, and R. spinosissima) was estimated on high performance thin layer chromatography cellulose plates. Differences in flavanol and proanthocyanidin profiles of the extracts were evident, among which Rosa spinosissima stood out with catechin as the only detected flavanol and red zones as indication of anthocyanins. Furthermore, the elution solvent for thin layer chromatography with mass spectrometry analyses of glycosylated flavan-3-ols and proanthocyanidins was optimized, enabling identification of catechin, (epi)catechin hexoside, proanthocyanidin dimer, and proanthocyanidin dimers and trimers hexosides. A total of 15 flavanols and their derivatives were identified using ultra-high-performance liquid chromatography with linear trap quadrupole-Orbitrap mass analyzer and epicatechin, gallocatechin, and proanthocyanidin trimer were identified only using this technique. However, proanthocyanidin trimer trihexoside was identified only by thin-layer chromatography with mass spectrometry. To establish the relationships between the flavanols and proanthocyanidins composition of rose hip and their origin, principal component analysis was performed on the entire set of liquid chromatography/mass spectrometry data. Both principal components' scores plots showed that Rosa spinosissima could be considered as an outlier. Our study demonstrated that flavanol and proanthocyanidin profiles of different rose hips depend on the geographical origin rather than on the cultivar and genotype.

Chemometric Characterization of Strawberries and Blueberries according to Their Phenolic Profile: Combined Effect of Cultivar and Cultivation System.

Chemical characterizations of leaves and fruits that were obtained from organically and integrally produced strawberries ('Favette', 'Alba', and 'Clery') and blueberries ('Bluecrop', 'Duke', and 'Nui') from western Serbia were undertaken in this study. Phenolic analysis was done while using ultra-high performance liquid chromatography coupled to a linear ion trap-Orbitrap hybrid mass analyzer, while total phenolic content (TPC), total anthocyanin content (TAC), and radical-scavenging activity (RSA) by spectrophotometry. In general, leaves and fruits from blueberry showed higher levels of TPC and TAC as compared to strawberry. These chemical traits were larger in organic grown fruits and larger in leaves than fruits. The most abundant phenolics in leaves and fruits of blueberry was 5-O-caffeoylquinic acid, followed by quercetin 3-O-galactoside, while catechin, quercetin, and kaempferol 3-O-glucosid were dominant in the leaves and fruits of strawberry. cis, trans-Abscisic acid was detected in all fruit samples, but not in leaves. Blueberries (both fruits and leaves) were separated from strawberries, but only organic blueberry fruits were distinguished from integrated fruits, according to principal component analysis. Quercetin, kaempferol, 5-O-caffeoylquinic acid, ferulic acid, caffeic acid, catechin, p-coumaric acid, and p-hydroxybenzoic acid were the most influential phenolic compounds for the separation. Much higher contents of TPC, RSA, TAC, quercetin 3-O-galactoside, and quercetin were found in fruits and TPC, RSA, catechin, p-hydroxybenzoicacid, p-coumaricacid, and ferulic acid in leaves in all three blueberry cultivars and the strawberry cultivar 'Clery'. These phenolic compounds are good sources of antioxidant compounds with potentially high beneficial effects on human health.

Characterization of Sugar and Polyphenolic Diversity in Floral Nectar of Different 'Oblacinska' Sour Cherry Clones.

'Oblacinska' sour cherry, an autochthonous cultivar, is the most planted cultivar in Serbian orchards. Since fruit trees in temperate zone reward insects by producing nectar which 'quality' affects the efficiency of insect pollination, the aim of this study was analyzing of sugars and polyphenolics in floral nectar of 16 'Oblacinska' sour cherry clones with different yielding potential. The contents of sugars and sugar alcohols were analyzed by ion chromatography, while polyphenolic profile was established using liquid chromatography/mass spectrometry technique. Fourteen sugars and six sugar alcohols were detected in nectar samples and the most abundant were fructose, glucose, and sucrose. Eleven polyphenols were quantified using available standards, while another 17 were identified according to their exact masses and characteristic fragmentations. Among quantified polyphenols, rutin, naringenin, and chrysin were the most abundant in nectar. Principal component analysis showed that some polyphenol components (naringin, naringenin, and rutin) together with sugars had high impact of spatial distribution of nectar samples on score plot.

Valorisation of Raspberry Seeds in Cosmetic Industry-Green Solutions.

The fruit processing industry generates large quantities of by-products well known to be rich in bioactive compounds with numerous nutritional properties and beneficial effects for human health. We developed a strategy to valorise raspberry seeds and obtain valuable ingredients with potential application in cosmetic skincare formulas. Cold press extraction technology was applied to extract oil, and the remaining defatted raspberry seed cake was treated with three proline based deep eutectic solvents (DES) to extract polyphenols. The most potent was proline/citric acid extract, with free and total ellagic acid content (52.4 mg/L and 86.4 mg/L), total phenolic content (TPC, 550.1 mg GAE/L) and radical scavenging activity (RSA, 4742.7 mmol TE/L). After the direct mixing of the extract and after encapsulation with starch as a carrier, the skincare emulsion and microemulsion were characterised by irritation potential (Zein test), transepidermal water loss (TEWL), red blood cell (RBC), and DPPH antioxidant test. The resulting preparations were of improved quality in comparison to the control hand cream, with a low skin irritation effect, lower TEWL, and higher antioxidant potential. This work complies with circular economy principles and green technology standards, and represents the efficient model on how to reuse natural resources through waste minimization.

Profiling of Metabolites in Organically Grown Plums from Norway: Does Location or Cultivar Matter?

The aim of this work was to investigate the influence of two locations and seven cultivars on the profiling of metabolites in organically grown plums (Prunus domestica L.) fruit in Norway. P, K, and Ca were most abundant in the studied fruits, while Ba and Sr formed a clear line between the locations. The most abundant sugars were glucose, fructose, sucrose, and sorbitol, which together accounted for up to 97.00%. Quinic acid and malic acid were the predominant organic acids, while chlorogenic acid, rutin, and kaempferol-3-O-glucoside were the most abundant polyphenols. Plums from Ullensvang were characterized by a higher content of minerals, sugars, organic acids, total polyphenol content (TPC), and radical scavenging activity (RSA), while plums from Telemark had a higher content of quantified polyphenols. The cultivar 'Mallard' had the highest mineral and radical scavenging activity, 'Opal' had the sweetest fruit, 'Jubileum' had the highest acidity, 'Excalibur' had the highest TPC content, and 'Valor' stored the highest content of quantified polyphenols, especially chlorogenic acid. These results provide comprehensive information on the chemical profiles of selected plum cultivars, suggesting that organic plums are a rich source of beneficial compounds that can have a positive impact on human health.

Fruit Quality Attributes of Organically Grown Norwegian Apples Are Affected by Cultivar and Location.

In this work, 12 apple cultivars grown organically in three regions of Norway (Telemark, Ullensvang, Viken) were analyzed in terms of fruit quality, with the aim of equating different growing regions under specific climatic conditions. Apples were analyzed for concentration levels of minerals, sugars, sugar alcohols, organic acids, total phenolic content (TPC), radical scavenging activity (RSA), and phenolic profiles. Discovery "Rose" from Telemark stored the highest level of minerals (24,094.5 mg/kg dry weight). Glucose, fructose, sucrose, and sorbitol were the major carbohydrates, whereas the predominant organic acids were quinic acid and malic acid. Cultivar Discovery from Ullensvang had the highest TPC (9.22 g/kg) and RSA (229.32 mmol TE/kg). Of the polyphenols quantified, chlorogenic acid and kaempferol-3-O-glucoside were the most abounded, accounting for 85.50%. Principal component analysis (PCA) shows that the Ullensvang region is the richest source of most carbohydrates, organic acids (quinic, shikimic, and galacturonic), and most polyphenols, whereas the highest content of minerals and maleic acid characterized Viken. Regardless of location, the Discovery cultivar had, on average, the highest sugar and polyphenol contents. The results obtained suggest that organic apples from Norway are a rich source of beneficial compounds that can have a positive impact on human health. In addition, these results may be useful for consumers in identifying apple cultivars with desirable characteristics and for the fruit industry in tracing back the origin of apples. The findings could also be of great interest for locations with similar climate and soil conditions worldwide.

Does Shoot Age Influence Biological and Chemical Properties in Black Currant (Ribes nigrum L.) Cultivars?

The aim of this study was to examine the influence of shoot age on the biological and chemical properties of 13 black currant cultivars with different origins and ripening times. Phenological observations together with examined pomological and chemical characteristics were studied in two consecutive years at the experimental field near Belgrade, Serbia. The total content of phenols was estimated spectrophotometrically by the Folin-Ciocalteu method, while quantitative analysis of anthocyanin and flavonols aglycones was performed using a high-performance liquid chromatographic (HPLC) method. Principal component analysis was performed to establish differences in biological and chemical properties of black currants. Three-year-old shoots had an earlier start of all examined phenological stages, better generative potential, higher yields, while clusters and berries from 2-year-old shoots had significantly higher values for physical properties, total phenols, anthocyanin and flavanols aglycones and antiradical capacity. Late ripening cultivars had higher contents of all chemical compounds. The berries on 2-year-old shoots had total phenolics that ranged between 123.0 ('Titania') and 298.3 mg/100 g fresh weight (FW) ('Ometa'), while total anthocyanins ranged between 398.5 ('Ojebyn') and 1160.8 mg/kg FW ('Ometa'). According to the obtained results, cultivars 'Ometa', 'Ben Lomond', 'Tsema' and 'Malling Juel' can be recommended as the most promising for growing in the continental climate because they stood out with higher generative potential and yield, physical traits of cluster and berry, higher level of primary and secondary metabolites and DPPH activity in their berries.

When Is the Right Moment to Pick Blueberries? Variation in Agronomic and Chemical Properties of Blueberry (Vaccinium corymbosum) Cultivars at Different Harvest Times.

Blueberries, which are recognized by their colored fruits and exquisite flavor and taste, are a great source of bioactive substances with potential functional properties. For the purpose of this study, the blueberry cultivars 'Duke', 'Chandler' and 'Bluecrop' were picked at four different times. The aim of the study was to compare the cultivars and determine the best time for picking fruits for table consumption and to produce berries that can be used as functional foods with elevated levels of bioactive compounds. According to principal component analysis (PCA), the most influential traits for distinguishing different times of harvest in the 'Duke' cultivar were sorbitol, glucose, sucrose, and turanose; for the cultivar 'Chandler', they were caffeic acid, aesculetin, and quercetin; for the 'Bluecrop', they were fructose, maltose, radical scavenging activity, and quercetin. Blueberry fruits aimed for table consumption were those harvested in the first two pickings of the cultivar 'Duke', in the first and third of the 'Bluecrop', and in the third picking time of the cultivar 'Chandler', due to the highest fruit size and very high level of sugar (mostly glucose and fructose). 'Duke' berries from the second and third harvest (high level of total phenolic content, radical scavenging activity, total anthocyanins, aesculin, quercetin, and isorhamnetin), 'Chandler' from the first and third (the highest p-hydroxybenzoic acid, aesculetin, caffeic acid, phloridzin, kaempferol, kaempferol 3-O-glucoside, quercetin 3-O-rhamnoside, rutin, and quercetin) and 'Bluecrop' from the third harvest (highest level of total phenolics, radical scavenging activity, quercetin, rutin, quercetin 3-O-glucoside, kaempferol, quercetin 3-O-rhamnoside, kaempferol 3-O-glucoside, and isorhamnetin) had the highest levels of health-promoting compounds.

Influence of extraction time, solvent and wood specie on experimentally aged spirits - A simple tool to differentiate wood species used in cooperage.

Type of the wood used for the aging highly influences the quality of alcoholic beverages. In this research we explored the potential of cyclic voltammetry (CV) and differential pulse voltammetry (DPV) to establish fingerprints characteristic for each wood and to enable determining the type of the wood used in the aging process. Eleven different wood samples were used to prepare three different types of spirits during 15 months. The highest extraction rate was obtained during the first month, while further aging was followed with almost constant amount of extracted polyphenols. Black locust, myrobalan plum, and mulberry extracts were discriminated from the spirits aged in oak and wild cherry wood when statistical analysis was applied. Although clear classification of all samples was not achieved, this long term study demonstrated a potential of both CV and DPV for differentiating wood species used in the aging, hence in the quality control of spirits.

Analysis of Phenolic Compounds for the Determination of Grafts (in) Compatibility Using In Vitro Callus Cultures of Sato-Zakura Cherries.

The aim of this study was to prove that under in vitro conditions, the adhesiveness of the callus between rootstock and scion, the development of callus cells at the points of fusion, and the presence of phenolic components are closely related to the level of (in) compatibility of the grafting combinations between Sato-zakura cherry cultivars ('Amanogawa', 'Kanzan', and 'Kiku-shidare-zakura') and commercial rootstocks. Prunus avium, Prunus 'Colt', Prunus mahaleb and Prunus serrulata were used as compatible and Prunus serotina and Pyrus communis 'Pyrodwarf' were used as two potentially incompatible rootstocks. The results indicated the significant manifestations of the early signs of the incompatibility on the callus junction. Phenols, as well as tissue senescence, were very precisely localized by toluidine blue and alcian blue as well as safranin staining, which can indicate the early signs of the callus incompatibility in some grafting unions. In the callus unions of Prunus avium with 'Amanogawa' and 'Kiku-shidare-zakura' the results of chemical analyses indicated that the existence of several flavonols, flavones and phenol acids could be involved in the incompatibility process in grafted combination. The detection of flavonol astragalin in the unions can be a biomarker of compatibility between scion and the rootstock, while some polyphenols, such as neochlorogenic acid, sinapic acid, ellagic acid, caffeic acid, baicalein, naringenin, apigenin and luteolin can be used as the indicators of graft incompatibility. p-coumaric acid and ferulic acid could be used for detection of delayed incompatibility.

Release of wood extractable elements in experimental spirit model: Health risk assessment of the wood species generated in Balkan cooperage.

This work investigates the release of toxic elements from wood into the experimental spirit models and the safety risks for consumers. The spirit models were prepared as ethanolic extracts using the procedure which reproduces maturation of spirits. Investigation included staves of wood species commonly used in Balkan cooperage: mulberry, Myrobalan plum, black locust, wild cherry, and various oaks. Potassium was the most abundant element, except in the wild cherry extract where calcium was dominant, and the Myrobalan plum extract where phosphorus was the most abundant. The parameters for the health risk assessment, such as hazard index (HI) and hazard quotient (HQ) were calculated for potentially toxic elements and indicated that all wood extracts would be safe for human consumption. Owing to the proven abundance of phenolics in the investigated wood extracts, relations among elements and phenolics were also studied and conclusions were made based on the statistically significant correlations.

Radical Scavenging and Antimicrobial Properties of Polyphenol Rich Waste Wood Extracts.

The main focus of this study is to assess radical scavenging and antimicrobial activities of the 11 wood extracts: oak (Quercus petraea (Matt.) Liebl., Q. robur L., and Q. cerris L.), mulberry (Morus alba L.), myrobalan plum (Prunus cerasifera Ehrh.), black locust (Robinia pseudoacacia L.), and wild cherry (Prunus avium L.). High-performance thin-layer chromatography (HPTLC) provided initial phenolic screening and revealed different chemical patterns among investigated wood extracts. To identify individual compounds with radical scavenging activity DPPH-HPTLC, assay was applied. Gallic acid, ferulic and/or caffeic acids were identified as the compounds with the highest contribution of total radical scavenging activity. Principal component analysis was applied on the data set obtained from HPTLC chromatogram to classify samples based on chemical fingerprints: Quercus spp. formed separate clusters from the other wood samples. The wood extracts were evaluated for their antimicrobial activity against eight representative human and opportunistic pathogens. The lowest minimum inhibitory concentration (MIC) was recorded against Staphylococcus aureus for black locust, cherry and mulberry wood extracts. This work provided simple, low-cost and high-throughput screening of phenolic compounds and assessments of the radical scavenging properties of selected individual metabolites from natural matrix that contributed to scavenge free radicals.

Chemical Fingerprint of 'Oblacinska' Sour Cherry (Prunuscerasus L.) Pollen.

The aim of this research was to analyze sugars and phenolics of pollen obtained from 15 different 'Oblacinska' sour cherry clones and to assess the chemical fingerprint of this cultivar. Carbohydrate analysis was done using high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD), while polyphenols were analyzed by ultra-high-performance liquid chromatography-diode array detector-tandem mass spectrometry (UHPLC-DAD MS/MS) system. Glucose was the most abundant sugar, followed by fructose and sucrose. Some samples had high level of stress sugars, especially trehalose. Rutin was predominantly polyphenol in a quantity up to 181.12 mg/kg (clone III/9), with chlorogenic acid (up to 59.93 mg/kg in clone III/9) and p-coumaric acid (up to 53.99 mg/kg in clone VIII/1) coming after. According to the principal component analysis (PCA), fructose, maltose, maltotriose, sorbitol, and trehalose were the most important sugars in separating pollen samples. PCA showed splitting off clones VIII/1, IV/8, III/9, and V/P according to the quantity of phenolics and dissimilar profiles. Large differences in chemical composition of studied 'Oblacinska sour cherry' clone pollen were shown, proving that it is not a cultivar, but population. Finally, due to the highest level of phenolics, clones IV/8, XV/3, and VIII/1 could be singled out as a promising one for producing functional food and/or in medicinal treatments.

Phenolic profiles of Serbian autochthonous variety 'Prokupac' and monovarietal international wines from the Central Serbia wine region.

Phenolic profiles of young monovarietal white and red wines from the Central Serbia were investigated. Quantification of phenolics was done using ultra-high-performance liquid chromatography (UHPLC) coupled with a diode array detector and a triple-quadrupole mass spectrometer. Generally, red wines were characterised with higher phenolics content when compared to the white wines. This applies also for the total phenolics content (TPC) and radical scavenging activity (RSA) values. Among all the samples, the red wine from autochthonous Serbian variety 'Prokupac' stood out with unique composition, showing the highest TPC, RSA and the highest concentration of the individual phenolics. Finally, based on a specific phenolic pattern, principal component analysis (PCA) indicated clustering of red and white wines, with the 'Prokupac' as a prominent outlier.

Carbon stable isotope composition of modern and archaeological Cornelian cherry fruit stones: a pilot study.

The carbon stable isotope content of Cornelian cherry stones collected from wild tree stands in Serbia, SE Europe, was measured using elemental analyser-isotope ratio mass spectrometry, with the aim of recording natural carbon isotope composition of the fruit stones and its possible variation. The results show a significant variation in the carbon isotope values; we identified several environmental factors that, along with a number of other possible determinants, likely contributed to this variation. The obtained data are compared with the measurement of carbon isotope content of an archaeological specimen of Cornelian cherry stone discovered at the Neolithic site of Vinca (ca. 5600-4500 BC) in Serbia. Notwithstanding the limitedness of the data and the complexity surrounding carbon fractionation and the isotopic variation, it is suggested that the differences/similarities in carbon isotope ratios between modern and archaeological Cornelian cherry stones, when measured for much larger assemblages, could potentially offer a glimpse into growing conditions of Cornelian cherry trees in the past.

Fatty acid and phenolic profiles of almond grown in Serbia.

Almond production is not typical for Serbia however the existence of natural populations and unexpectedly suitable agro-climatic conditions initiated this kind of study. Total oil content and concentrations of the fatty acids, total phenolic content and radical-scavenging activity were determined in the kernel oil of 20 local almond selections originating from North Serbia and cultivars 'Marcona', 'Texas' and 'Troito'. Sixteen fatty acids were identified and quantified, with the most abundant being oleic acid and linoleic acid. Nine phenolic acids and nineteen flavonoids were quantified using UHPLC-DAD MS/MS. The predominant polyphenol was catechin, followed by chlorogenic acid and naringenin. Based on oleic acid/linoleic acid ratio, levels of unsaturated fatty acids and specific polyphenols, some selections were chosen for growing and could also be recommended for breeding programs. Our investigation demonstrated that this region could be a suitable for growing almonds with chemical compositions competitive with standard cultivars.

Impact of Clonal Variability on Phenolics and Radical Scavenging Activity of Grapes and Wines: A Study on the Recently Developed Merlot and Cabernet Franc Clones (Vitis vinifera L.).

In this study, grapes and corresponding wines of Merlot /No 022, 025 and 029/ and Cabernet Franc /No 02, 010 and 012/ clones (recently developed) were evaluated regarding the total phenolic content, total anthocyanin content, and radical scavenging activity, aiming to better understand their quality and market potential. The nineteen individual polyphenols were quantified in studied grape samples using UHPLC coupled to a triple-quadrupole mass spectrometer. The mother grapes and wines were used as the relevant standards. In the grape, studied characteristics were monitored at three stages of berry development: green berry, véraison and mature berry. The mature grape of clones presented high values of total phenolics (3.81-10.89 g gallic acid equivalent kg-1 frozen weight), anthocyanin content (359.00-1668.18 mg malvidin-3-O-glucoside kg-1 frozen weight) and the radical scavenging activity (41.37-80.48 mmol trolox equivalent kg-1 frozen weight) depending on the clone. Grapes and wines of Merlot No 025 and Cabernet Franc No 010 stood out with the highest values of all three parameters. Generally, the high correlation was observed between TPC and RSA values for green berries and mature grapes, as well as for wine samples. The most abundant phenolics in both Merlot and Cabernet Franc grapes were gallic acid, p-hydroxybenzoic acid, ferulic acid, catechin, epicatechin, gallocatechin gallate, catechin gallate, and rutin. Catechin, epicatechin, epigallocatechin, and catechin gallate, typical for the ripe grape of Merlot and Cabernet Franc clones, showed significant correlation with RSA values. Concentrations of individual polyphenols varied depending on the sample being studied and berry development stages. Merlot No 025 and Cabernet Franc No 010 grapes showed particularly interesting attributes for the production of high quality wines.

Identification and quantification of phenolic compounds in berry skin, pulp, and seeds in 13 grapevine varieties grown in Serbia.

Phenolic profile of 13 grapevine varieties was determined, with respect to three different parts of berries: skin, seed and pulp. Identification and quantification of the phenols was done using ultra-high performance liquid chromatography (UHPLC) coupled with a diode array detector and a triple-quadrupole mass spectrometer. The most abundant phenols in grape seeds were found to be flavan-3-ols, most of which are gallocatechin gallate and catechin. The skins were characterized mostly by flavonols, i.e. quercetin and myricetin. Characterization of anthocyanins in the berry skin by UHPLC coupled with Linear Trap Quadrupole and OrbiTrap mass analyzer revealed a total of twenty derivatives of malvidin, delphinidin, petunidin, cyanidin, and peonidin. To the extent of our knowledge this is the first work that shows the presence of malvidin 3,5-O-dihexoside in the berry skin of 'Merlot', 'Cabernet Franc', 'Shiraz' 'Sangiovese', 'Pinot Noir' and 'Prokupac', untypical for Vitis vinifera Linneo species.