A detailed survey of agroecological status of Allium ursinum across the republic of Serbia: Mineral composition and bioaccumulation potential.

The purpose of this study was to determine the content of twenty-two biogenic elements (BEs) and potentially toxic elements (PTEs) in the soil and fresh Allium ursinum leaves from 43 different locations, in order to examine their bioaccumulation potential. Analyses of soil and plant material were carried out by using Inductively Coupled Plasma coupled with Optical Emission Spectroscopy (ICP-OES), a mercury analyzer (Hg), liquid chromatography (Cr), and AAS hybrid technique (As). The obtained results of the investigated elements were compared with the proposed limit values. The soil contamination factor (CF) as well as plant bioaccumulation factor (BAF) were calculated and the correlation analysis was performed. The results showed that the content of some BEs/PTEs in the soil were above the limit values, with two locations highly contaminated (CF > 6) with five (Cr(VI), Cu, Mn, Ni, V) and four (As, Co, Pb, V) elements. The content of As, Cd, Cr, and Pb in the leaves was higher than the permitted levels at some locations. The BAF was high (K, Ca, Zn, As), medium (Mg, Cu, B, Ni, Na, Pb), and low (Fe, Mn, Cr). The correlation between BEs/PTEs content in the leaves and soil was not significant, except for the following elements: Cd (0.37), Ca (0.34), As (0.36), Pb (0.30), and Na (0.25). The observed medium correlation suggested that the detected elements originated both from the atmosphere and the soil. Although A. ursinum at examined locations seemed to be mostly safe for consumption, a selective mechanism of adoption of certain BEs/PTEs requires continuous monitoring of their content in the future, to avoid quantities that can jeopardize human health through its consumption.

Authenticity assessment of cultivated berries via phenolic profiles of seeds.

Considering the health-benefits of berry fruits consumption and increased market demands for food authenticity as one of the most important quality assurances, phenolic profiling by high-performance thin layer chromatography and ultra-high-performance liquid chromatography hyphenated with mass spectrometry was combined with multivariate analysis for phytochemical characterization and intercultivar discrimination of cultivated berry seeds. The phenolic profiles of 45 berry seeds from nine genuine Serbian cultivated fruit species (strawberry, raspberry, blackberry, black currant, blueberry, gooseberry, cape gooseberry, chokeberry, and goji berry) revealed a good differentiation according to botanical origin. In order to determine biomarkers responsible for the classification, a total of 103 phenolic compounds were identified, including 53 phenolic acids and their derivatives, 26 flavonoids and 24 glycosides. Biomarkers derived from the phenolic profile of berry seeds proved to be a powerful tool in the authentication of botanical origin, and may be useful in detection of frauds in berry-based seed-containing product.

Primary Metabolite Chromatographic Profiling as a Tool for Chemotaxonomic Classification of Seeds from Berry Fruits.

Research background: Considering the importance of consumption of berry fruits with proven health-beneficial properties and difficulties in quality control of products of specific botanical and geographic origin, a fingerprint method was developed, based on advanced data analysis (pattern recognition, classification), in order to relate the variability of nutrients in the selected cultivars to primary metabolite profile. Experimental approach: Forty-five samples of genuine berry fruit cultivars (strawberry, raspberry, blackberry, black currant, blueberry, gooseberry, chokeberry, cape gooseberry and goji berry) were characterized according to chromatographic profiles of primary metabolites (sugars, lipids and fatty acids) obtained by three chromatographic techniques (high-performance thin-layer chromatography, gas chromatography coupled to mass spectrometry, and high-performance anion-exchange chromatography with pulsed amperometric detection). Results and conclusions: Comprehensive analysis allowed monitoring and identification of metabolites belonging to polar lipids, mono-, di- and triacylglycerols, free fatty acids, free sterols, sterol esters, mono- to heptasaccharides and sugar alcohols. Chemical fingerprint of berry seeds showed the uniformity of primary metabolites within each fruit species, but revealed differences depending on the botanical origin. All three chromatographic methods provided a discriminative, informative and predictive metabolomics methodology, which proved to be useful for chemotaxonomic classification. Novelty and scientific contribution: A novel methodology for the identification of bioactive compounds from primary metabolites of natural products was described. The proposed untargeted metabolite profiling approach could be used in the future as a routine method for tracing of novel bioactive compounds. The knowledge of metabolite composition obtained in this study can provide a better assessment of genotypic and phenotypic differences between berry fruit species and varieties, and could contribute to the development of new breeding programs.

Effect-Directed Profiling of Strawberry Varieties and Breeding Materials via Planar Chromatography and Chemometrics.

Strawberries are an important fruit in the European diet because of their unique taste and high content of essential nutrients and bioactive compounds. The anthocyanins are known to be colorful phenolics in strawberries. In 17 samples of six strawberry cultivars produced in Serbia, i.e., the common varieties Alba, Asia, and Clery as well as promising breeding materials (11.29.11, 11.34.6, and 11.39.3), the anthocyanin profile as well as antimicrobial and antioxidative activity profiles were determined. All investigated extracts showed antioxidative and antibacterial activities against Gram-negative Aliivibrio fischeri. The responses were quite similar in number and intensity. The HPTLC-DPPH• scavenging assay and HPTLC-Aliivibrio fischeri bioassay coupled with high-resolution mass spectrometry identified pelargonidin-3-O-glucoside (Pg-3-glc) as the main anthocyanin and prominent antioxidative and antimicrobial compound in strawberries. The density functional theory calculations at the M06-2X/6-31+G(d,p) level showed that Pg-3-glc quenches free radicals via sequential proton loss electron transfer mechanism in water and in pentyl ethanoate, where the 5-OH group is the most reactive site for proton and hydrogen atom transfer. The results were confirmed via spectrophotometry. The highest total phenolic content was found in Clery and 11.39.3, while statistically significant differences between the genotypes regarding the antioxidant activity were not confirmed. Although very similar in the anthocyanin, antioxidative, and antimicrobial profile patterns, the strawberry genotypes were successfully classified using principal component analysis.

Quality Assessment of Apple and Grape Juices from Serbian and German Markets by Planar Chromatography-Chemometrics.

The high consumption of plant-based foods on a global scale has increased the number of adulterations in the food industry. Along with this, analytical approaches to fraud detection need to be further developed. A nontargeted effect-directed profiling by high-performance thin-layer chromatography hyphenated with five effect-directed assays (free radical scavenging assay, Aliivibrio fischeri bioassay, and acetylcholinesterase, butyrylcholinesterase, and tyrosinase inhibition assays) and multi-imaging provided additional information on the antioxidative, antimicrobial, and enzyme inhibition activities for 18 apple and 18 grape juices from markets in Serbia and Germany. Bioactive zones of interest were eluted using an elution head-based interface and further characterized by electrospray ionization high-resolution mass spectrometry. The different profiles were evaluated chemometrically, and several compounds, which were characteristic of samples from different markets located in Serbia and Germany, were identified in apple juice (such as chlorogenic acid, phloridzin, epicatechin, and caffeic acid) and grape juice (such as chlorogenic acid, epicatechin, and quercetin). The developed rapid and simple method for the quality assessment of fruit juices coming from different (geographic) markets showed clear quality differences. Thus, it could be used to learn more about quality differences, to detect fraud in fruit juice production, and to verify the authenticity of the origin.

Comprehensive electrophoretic profiling of proteins as a powerful tool for authenticity assessment of seeds of cultivated berry fruits.

Product authentication is one of the most important food quality assurances. Considering the importance of consumption of berry fruits with proven health-beneficial properties, high sensory values and rich composition in bioactive substances, the aim of this study was to evaluate a straightforward and simple procedure for the protein fingerprinting of berry seeds. For this purpose, protein profiles of 45 samples of genuine berry fruit cultivars (strawberry, raspberry, blackberry, black currant, blueberry, gooseberry, chokeberry, cape gooseberry, and gojiberry) were analyzed by SDS-PAGE electrophoresis in combination with advanced chemometric tools. The most important parameters for discrimination among berry seeds were polypeptides at 12.8; 15.1; 25.0; 26.4; 30.0; 41.8; 44.4; 46.0; 48.5; 52.3 and 56.4 kDa. Biomarkers obtained from the protein profile of berry seeds proved to be a powerful tool in the authentication of their botanical origin, as well as for potential detection of berry-based products adulteration.

Organically vs. Conventionally Grown Vegetables: Multi-elemental Analysis and Nutritional Evaluation.

Vegetables are important contributors to a healthy diet, and their adequate daily intake can help prevent some of the major illnesses. The aim of the study was to examine the content of the major and trace elements in selected organically grown (OG) and conventionally grown (CG) vegetables (cabbage, kohlrabi, Brussels sprout, beetroot, carrot, potato, and onion), taken from city green markets. Multi-elemental analysis was carried out by inductively coupled plasma method with optical emission spectrometry (ICP-OES). Nutritional quality evaluation in comparison to nutritional reference values was done. In studied vegetables, Al, Ca, K, Fe (with the exception of organic kohlrabi), Mg, Na, P, S, and Zn were quantified in all samples, whereas As, Cd, Co, Hg, Se, and V were below the limit of detection for these elements. Macroelements and trace elements were found at higher concentrations in OG and CG vegetables, respectively. Differences in concentrations of studied elements between the same vegetable species produced in two agricultural systems were significant, except for beetroot (p ≤ 0.05). Principal component analysis and hierarchical cluster analysis results showed that the botanical origin had higher influence on sample differentiation than the agronomic practice, which was in accordance with the results obtained by Mann-Whitney U test. Good quality of both OG and CG vegetables in respect of nutritionally beneficial elements was observed.

Circulatory Imbalance of Essential and Toxic Trace Elements in Pre-dialysis and Hemodialysis Patients.

The status of essential and toxic trace elements in patients with different stages of chronic kidney disease (CKD) is still unclear and not well characterized. The present study examined the circulatory levels of a wide panel of trace elements (Al, Cr, Mn, Co, Ni, Cu, Zn, As, Se, Rb, Sr, Cd, Pb, and U) in hemodialysis patients (HD group) and pre-dialysis patients with stage 3 CKD (PD group). Comparisons were made between groups of patients and healthy individuals from the control group (CG). The levels of Al, Mn, Co, Ni, Cu, As, Se, Sr, and Pb were higher, while the levels of Cr, Zn, Rb, Cd, and U were lower in HD patients than in our CG. Higher levels of Al and Se, as well as lower levels of As, Sr, Zn, Rb, and U were significant and distinguished HD from PD. Among other analyzed elements, Co, Se, and U are the only trace elements that did not distinguish PD from CG at a statistically significant level. The HD group had lower serum U levels than the PD group, and this could be a result of hemodialysis. This study also revealed that the Cu/Zn ratio could be used as a marker for early and late detection of renal failure. Marked changes of essential and toxic trace element levels in sera indicate additional pathophysiological events in CKD, which could additionally contribute to the preexisting increased morbidity of HD patients. Measurement of trace elements in HD patients should be performed routinely.

Alteration of Trace Elements in Multinodular Goiter, Thyroid Adenoma, and Thyroid Cancer.

Modest progress has been made in understanding the role of trace elements as endocrine disruptors. The aim of this study was to examine whether there is a change in the content of trace elements in thyroid disease, as well as whether the ratio of elements could be considered a blood marker for thyroid disease. In addition, this study examined the influence of biological and clinical/pathological parameters on the elemental profile. Blood samples from patients diagnosed with multinodular goiter (MNG), thyroid adenoma (TA), and thyroid cancer (TC) were examined and compared with control samples using chemometric analysis. The concentrations of essential (Mn, Co, Cu, Zn, Se) and toxic elements (Ni, As, Cd, Pb, U) were determined by ICP-MS. This study showed for the first time that the content of Mn, Co, Ni, Cu, Zn, Se, and Pb in pathological blood samples was significantly lower compared to the control, while opposite results were obtained for As, Cd, and U. Based on the classification model, the most important trace metals for discrimination of MNG and TC from the control group (CG) were Co and Zn, while Co, Zn, and Mn influenced the distinction of CG from TA. Moreover, it was found that Cu/Zn and U/Se ratios had significantly increased values in pathological blood samples leading to the possibility of establishing new circulating screening markers. These findings can represent significant translational information since these diseases are widespread and the diagnostic procedure is still difficult in many cases.

Multi-elemental Analysis, Pattern Recognition Techniques of Wild and Cultivated Rosehips from Serbia, and Nutritional Aspect.

Twenty-six major and trace elements, in the seed and the mesocarp of wild and cultivated rosehips from different locations in Serbia, were quantified by means of inductively coupled plasma-atomic emission spectroscopy (ICP-AES). The cultivated rosehip was investigated for the first time. In both wild and cultivated rosehips, the most abundant elements were K and Ca. Among trace elements, Mn (in both seed and mesocarp); Fe (in seeds); and B, Ba, and Sr (in mesocarp) were quantified in the highest concentrations. The higher content of Cu, K, Mn, P, and S in the seed of cultivated rosehip, as well as Ca, Mg, and Sr in the mesocarp of wild rosehip, was observed, both significant at p < 0.05. Additionally, differences between the seed and the mesocarp of studied rosehip were noticed in the content of B, Cu, Fe, K, Ni, P, S, Sr, and Ti (p < 0.05). Nutritional assessment revealed that both wild and cultivated rosehips are a valuable source of essential elements (Ca, Cu, K, Mg, Mn, and P). The absence of toxic and potentially toxic elements additionally contributes to the quality of studied Serbian rosehip. A high impact of factors such as variety, location, as well as their interaction on the content of elements in cultivated rosehip mesocarp and seed was observed. Pattern recognition techniques, principal component analysis (PCA), and hierarchical cluster analysis (HCA) were applied in order to provide insight into similarities among the analyzed samples.

Evaluation of trace metals in thyroid tissues: Comparative analysis with benign and malignant thyroid diseases.

Evaluation of trace metals at level of solid tissue can provide better information than blood or urine and, therefore, could highlight the role of metals in the etiology of organ-specific disease. The current study aimed to establish the baseline content of four essential (Mn, Cu, Zn, Se) and four toxic metals (As, Cd, Pb, U) in the healthy thyroid tissues (HTTs) by considering sex, age and smoking habits. A further aim was to examine whether differences in the content of metals exist in regard to the thyroid diseases, such as benign tumor (BT), Hashimoto's thyroiditis (HT), multinodular goiter (MNG) and thyroid cancer (TC). A total number of investigated tissue samples were 423. All metals were quantified by inductively coupled plasma-mass spectrometry (ICP-MS). It was found that the content of Cu and U was higher in HTTs of women, while the content of Zn was higher in HTTs of men. Increased content of Zn and decreased content of U was found in the group of HTTs above 50 years compared to a younger group (<50 years). Increased content of Cd, Pb and U distinguish smokers from the non-smokers. In comparison with other population groups worldwide, investigated Serbian population had up to 15 times reduced content of Se. Despite the difference in metal's profile according to biological variables, this study also demonstrated, for the first time, that each thyroid disease has its unique metal's profile. The most altered metal's content was found in tissues with HT. Contrarily, the greatest similarity in metal's content with HTTs was found in BT tissues. Based on the increased content, metal's that dominantly discriminated HTTs from the HT, MNG and TC was As, Pb and Cd, respectively. Reported results could highlight the role of toxic and essential trace metals in the not very well clarified etiology of thyroid diseases and, moreover, could provide a molecular basis for pathophysiological changes of metal's hazardous effects on thyroid health at the tissue level.

Cadmium as main endocrine disruptor in papillary thyroid carcinoma and the significance of Cd/Se ratio for thyroid tissue pathophysiology.

BACKGROUND: The etiology of papillary thyroid carcinoma (PTC) is unknown and some literature data support the hypothesis that heavy metals, as endocrine disrupters, could play a major role in the pathogenesis of thyroid cancer. This study aimed to estimate the content of selected toxic and essential trace metals (Mn, Co, Ni, Cu, Zn, As, Se, Cd, Pb, Th, and U), as well as the selected ratio's (Cu/Zn and Cd/Se) in the malignant thyroid tissues according to sex, age, smoking habits, familial history of any thyroid disease, pathohistological (PH) types of PTC, tumor size, the existence of a thyroid capsular invasion, intrathyroid tumor dissemination, retrosternal thyroid growth, and TNM progress of PTC. METHODS: The study included 66 patients with PTC (women/men ratio = 46/20, mean age: 54 ± 14 years). A comparative analysis was made by collecting the healthy thyroid tissues (HTTs) of the same patients, making the total number of samples 132. All trace metals were quantified by inductively coupled plasma-mass spectrometry (ICP-MS). RESULTS: Metals that significantly separated papillary thyroid tissues (PTTs) from the HTTs were Cd, U and Se (p < 0.05). The obtained negative correlation between Cd and Se in the PTTs could explain extrusion of essential Se caused by increased content of Cd. Only Cd had an influence on the retrosternal thyroid growth, while the essential metals (Mn, Co, and Zn) had an influence on thyroid capsular invasion. CONCLUSION: It was found that Cd act as the main endocrine disrupter, which could highlight its role in the etiology of PTC. Considering that the Cd/Se ratio significantly separated two studied groups and had an influence on the retrosternal thyroid growth, its altered content could contribute to the better understanding of the molecular basis for pathophysiological changes in the PTC.

Distribution of elements in seeds of some wild and cultivated fruits. Nutrition and authenticity aspects.

BACKGROUND: The compositional, functional, and nutritional properties of fruits are important for defining their quality. Fruit seeds should be better exploited as they are also considered to be a good source of bioactive components. Twenty macro, micro, and trace elements were identified and quantified in the seeds of 70 genuine wild and cultivated fruit species/cultivars by inductively coupled plasma atomic emission spectrometry and inductively coupled plasma mass spectrometry. Sophisticated chemometric techniques were also used to establish criteria for the classification of the analyzed samples. RESULTS: Calcium and P were the most abundant elements, followed by K and Na. The content of microelements and trace elements differed among the different cultivars/genotypes. The content of Ba, Pb, and Sr was significantly higher in wild fruits, whereas Fe, Mg, Mn, Ni and Zn content was higher in cultivated fruits. CONCLUSION: All of the statistical procedures that were used - Kruskal-Wallis, Mann-Whitney U-test, and principal component analysis (PCA) - confirm a unique set of parameters that could be used as phytochemical biomarkers to differentiate fruit-seed samples belonging to different cultivars/genotypes according to their botanical origin. This kind of investigation may contribute to intercultivar/genetic discrimination and may enhance the possibilities of acquiring a valuable authenticity factor. © 2018 Society of Chemical Industry.