Aleksandra Mišan

Development of a rapid resolution HPLC method for the separation and determination of 17 phenolic compounds in crude plant extracts

AbstractRapid Resolution HPLC/DAD method, on a 1.8 µm, 4.6×50 mm column, was developed to enable a rapid separation of a mixture of 17 compounds, which consisted of hydroxybenzoic acids, hydroxycinnamic acids, flavones, flavonols, flavanone, flavonol-glycoside and antraquinone, in a single run, within 22 minutes. The developed method is precise, accurate and sensitive enough for simultaneous quantitative evaluation of major compounds in crude and hydrolyzed extracts of parsley, buckthorn, mint, caraway and birch. In order to overcome the inability to quantify all the phenolic compounds present in the samples caused by lack of external standards, HPLC approaches for the total phenolic content estimation based on sum of all integrated peak areas were made. These results were compared with the total phenolic content determined by Folin-Cioacalteu method. Although the correlation between the series of data was not significant (p<0.05), the difference between the results of total phenolic content obtained spectrophotometrically and by HPLC was not high in the case of parsley, buckthorn and mint extract. Regarding the obtained results, the HPLC approach could serve as an excellent tool for total phenolic content estimation, without the need for complete identification of the individual compounds.

Buckwheat (Fagopyrum esculentum Moench) Grain and Fractions: Antioxidant Compounds and Activities

UNLABELLED Buckwheat (Fagopyrum esculentum Moench) is an alternative crop belonging to the Polygonaceae family. In comparison to antioxidant activity of frequently used cereals, buckwheat has been reported to possess higher antioxidant activity (AOA), mainly due to high rutin content. The objective of this work was to determine the main antioxidant compounds and AOA of buckwheat grain fractions (whole grain, hull, and groat). Buckwheat grain fractions were extracted with ethanol/water (80/20, v/v), followed by determination of total phenolic and flavonoid content. Quantification of phenolic compounds and tocopherols was performed by high-performance liquid chromatography (HPLC). The AOA was estimated by 2 direct electron spin resonance (ESR) and 4 indirect (spectrophotometric) tests. Significantly higher contents of total phenolics and total flavonoids were found in buckwheat hull than in whole grain and groat. Protocatechuic, syringic, and sinapic acid, rutin, and quercetin were found in all tested fractions, whereas vanilic acid was found in whole grain and hull. The content of total tocopherols in investigated samples ranged from 23.3 mmol/g for hull to 61.8 mmol/g for groat. Hull was superior in scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH(•)), hydroxyl ((•)OH), and superoxide anion (O(2)(•-)) radicals, reducing activity, AOA by β-carotene bleaching method, and chelating activity on Fe(2+) as evidenced by its lower IC(50) value. Obtained results can broaden the utilization of buckwheat, especially a share of hull in whole grain flour production. PRACTICAL APPLICATION Obtained results suggest possibility to supplement the whole grain buckwheat flour with hull, which leads toward better usage of by-products in buckwheat production, and enhancement of antioxidant potential of the final product.

Comparison of Antioxidant Components and Activity of Buckwheat and Wheat Flours

ABSTRACT Plant phenolics and tocopherols content were determined in light and wholegrain buckwheat and wheat flour. Antioxidant activity of flours were comparatively assessed by scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH•), hydroxyl (•OH), and superoxide anion (O2•–) radicals, reducing activity, and chelating activity on Fe2+. Rutin, quercetin, and ferulic acid were quantified in both buckwheat flours, while ferulic acid was quantified in wholegrain wheat flour. Significantly higher content of phenolics and tocopherols was found in buckwheat than in wheat flours. Tocopherols in buckwheat flours were present in the order: γ- > α->> δ-tocopherol, and in wheat flours: α- > γ- >> δ-tocopherol. Buckwheat flours possessed better scavenging abilities on DPPH•, •OH and O2•– radicals, as well as better reducing activity, while wheat flours showed better chelating activity on Fe2+, according to IC50 values. Results suggest the possibility of improving the antioxidant properties of wheat-based food p...

Characterization of Byproducts Originating from Hemp Oil Processing

Valorization of hemp seed meal, a byproduct of hemp oil processing, was performed by measuring the distribution of nutritional and antinutritional compounds in different hemp seed meal fractions. According to chemical composition, two cotyledon-containing fractions (>180 and <180 μm) were significantly richer in protein (p < 0.05) (41.2% ± 0.04% and 44.4% ± 0.02%, respectively), lipid (15.1% ± 0.02% and 18.6% ± 0.04%, respectively), and sugar content (4.96% ± 0.11% and 3.46% ± 0.08%, respectively) in comparison to the hull-containing fractions (>350 and >250 μm), which were significantly richer in crude fiber content (29.5% ± 0.04% and 21.3% ± 0.03%, respectively). The free radical scavenging capacity (IC50) of fraction extracts increased (p < 0.05) with increasing mean particle size (from 17.18 ± 0.59 to 5.29 ± 0.30 mg/mL). Cannabisin B and N-trans-caffeoyltyramine were the most abundant phenolic compounds in the hull fractions (from 267 ± 15.9 to 287 ± 23.1 mg/kg), while cotyledon fractions had higher content of catechin (from 313 ± 12.4 to 744 ± 22.2 mg/kg) and p-hydroxybenzoic acid (from 124 ± 6.47 to 129 ± 8.56 mg/kg (P < 0.05). Well-balanced ω-6 to ω-3 fatty acid ratio (3:1) was determined in all fractions. Antinutrients (trypsin inhibitors, phytic acid, glucosinolates, and condensed tannins) were mostly located in the cotyledon fractions. These findings indicate that the separation of hemp seed meal into different fractions could be used to concentrate valuable target compounds and consequently facilitate their recovery.

Phenolic Compounds Contained in Little-known Wild Fruits as Antiadhesive Agents Against the Beverage-Spoiling Bacteria Asaia spp.

The aim of the study was to evaluate antioxidant activity and total phenolic content of juice from three different types of fruits: elderberry (Sambucus nigra), lingonberry (Vaccinium vitis-idaea) and cornelian cherry (Cornus mas), and their action against adhesion of bacterial strains of Asaia lannensis and Asaia bogorensis isolated from spoiled soft drinks. The antioxidant profiles were determined by total antioxidant capacity (2,2-diphenyl-1-picrylhydrazyl, DPPH), and ferric-reducing antioxidant power (FRAP). Additionally, total polyphenol content (TPC) was investigated. Chemical compositions of juices were tested using the chromatographic techniques: high-performance liquid chromatography (HPLC) and liquid chromatography–mass spectrometry (LC-MS). Adhesion properties of Asaia spp. cells to various abiotic materials were evaluated by luminometry, plate count and fluorescence microscopy. Antioxidant activity of fruit juices expressed as inhibitory concentration (IC50) ranged from 0.042 ± 0.001 (cornelian cherry) to 0.021 ± 0.001 g/mL (elderberry). TPC ranged from 8.02 ± 0.027 (elderberry) to 2.33 ± 0.013 mg/mL (cornelian cherry). Cyanidin-3-sambubioside-5-glucoside, cyanidin-3-glucoside, and cyanidin-3-sambubioside were detected as the major anthocyanins and caffeic, cinnamic, gallic, protocatechuic, and p-coumaric acids as the major phenolic acids. A significant linear correlation was noted between TPC and antioxidant capacity. In the presence of fruit juices a significant decrease of bacterial adhesion from 74% (elderberry) to 67% (lingonberry) was observed. The high phenolic content indicated that these compounds may contribute to the reduction of Asaia spp. adhesion.

Determination of Volatile Organic Compounds in Selected Strains of Cyanobacteria

Microalgal biomass can be used in creating various functional food and feed products, but certain species of microalgae and cyanobacteria are known to produce various compounds causing off-flavour. In this work, we investigated selected cyanobacterial strains of Spirulina, Anabaena, and Nostoc genera originating from Serbia, with the aim of determining the chemical profile of volatile organic compounds produced by these organisms. Additionally, the influence of nitrogen level during growth on the production of volatile compounds was investigated for Nostoc and Anabaena strains. In addition, multivariate techniques, namely, principal component analysis (PCA) and hierarchical cluster analysis (HCA), were used for making distinction among different microalgal strains. The results show that the main volatile compounds in these species are medium chain length alkanes, but other odorous compounds such as 2-methylisoborneol (0.51–4.48%), 2-pentylfuran (0.72–8.98%), β-cyclocitral (0.00–1.17%), and β-ionone (1.15–2.72%) were also detected in the samples. Addition of nitrogen to growth medium was shown to negatively affect the production of 2-methylisoborneol, while geosmin was not detected in any of the analyzed samples, which indicates that the manipulation of growth conditions may be useful in reducing levels of some unwanted odor-causing components.

Antioxidant activity of wheat and buckwheat flours.

Antioxidative activities of wheat flours (type 500 and wholegrain) and buckwheat flours (light and wholegrain) were tested using 1,1-diphenyl-2-picrylhydrazyl (DPPH·)-scavenging activity, reducing power and chelating activity on Fe2+. Also, the content of the total phenolics of ethanolic extracts was estimated. Polyphenolics content (expressed as gallic acid equivalent, GAE) in wheat flours varied between 37.1 and 137.2 μg GAE/g extract, while its content in buckwheat flour were at least four time higher and ranged between 476.3 and 618.9μg GAE/g extract. Ethanolic extracts of buckwheat flours exhibited higher antioxidant activities in all the assays, except for chelating activity. Regarding all the obtained results, it can be concluded that bakery products produced with buckwheat flour could be regarded as potential functional foods.

Relationship of Physicochemical Characteristics with Sensory Profile of Cookies Enriched with Medicinal Herbs

In this work, a range of model cookies enriched with different quantities of medicinal herbs, applied in two forms (pulverized mixture and extract), were studied to compare and describe relationships between physicochemical and sensory data. Multivariate statistical methods were applied to estimate relationships in the analyzed data. Analysis of variance showed that the majority of analyzed parameters were highly significant in discriminating among the samples (p < 0.05), which supported the usefulness of their application in characterizing the quality profile of enriched cookies. Principal component analysis on physicochemical data revealed that cookies at the same enrichment level showed more similarities. Principal component analysis on sensory data exhibited a larger separation between the samples with increasing enrichment levels, especially in the case of pulverized herbal mixture. According to partial least squares regression, two significant partial least squares components explained 92.7% and predicted 78.0% variation in the data. The most important parameters in discriminating between the cookies enriched with medicinal herbs were lightness, red tonality, and hardness. The selected physicochemical parameters could be used in future studies to evaluate variously enriched cookies by establishing models and investigating the predictability of sensory quality.

Sugar beet molasses as an ingredient to enhance the nutritional and functional properties of gluten-free cookies.

Abstract Sugar beet molasses is a raw material with high potential to be a functional ingredient in baked goods. This paper investigated the nutritional and functional properties of gluten-free cookies enriched with sugar beet molasses. At all enrichment levels and forms tested (liquid and dry), the addition of beet molasses improved the micronutrient pattern and antioxidative status of gluten-free cookies. The cookies prepared with molasses were significantly higher in potassium, magnesium, calcium, iron, betaine, total phenolics and DPPH radical scavenging abilities. Molasses contributed to wider spectra of phenolic compounds. The dominating phenolic compounds in the molasses-enriched cookies were catechin, ferulic, syringic and vanillic acid. Molasses also contributed to the presence of p-hydroxybenzoic acid in the cookies. Addition of molasses increased the content of hydroxymethyfurfural in the cookies, but not above values commonly reported for this product type. Molasses addition improved the overall acceptance of gluten-free cookies up to 30% enrichment level.

Presence of aflatoxins in cereals from Serbia

Aflatoxins (AFs), one of the most toxic and the strongest natural carcinogens can be found in a variety of food commodities, including cereals. For that purpose, the aim of this study was to investigate occurrence of AFs (AFB1, AFG1, AFB2 and AFG2) in 130 cereal samples. AFs content was determined by direct competitive Enzyme Linked Immunosorbent Assay (ELISA) method. Samples with AFs content higher than 1 μg/kg were analyzed again with confirmatory High Performance Liquid Chromatography with fluorescence detection (HPLC-FLD). Analyses showed that none of the analyzed wheat (30), barley (20), oats (20) and rye (20) samples was contaminated with AFs. On the other hand, among 40 analyzed maize samples 24 of them (60%) were contaminated in the following way: 6 (25%) samples had AFs concentration between 1 and 10 μg/kg, 14 (58%) samples between 10 and 50 μg/kg and 4 (17%) between 50 and 70.3 μg/kg. The most predominant aflatoxin was AFB1 which was detected in all contaminated maize samples. AFG1, AFB2 and AFG2 were found in 12, 5 and 1 sample, respectively. This study represents the first investigation of the occurrence of AFs in five different cereals from Serbia.