How do in vitro digestion and cell metabolism affect the biological activity and phenolic profile of grape juice and wine.

Cabernet Sauvignon grape juice and wine underwent in vitro digestion, resulting in a reduction of most phenolic compounds (10%-100% decline), notably impacting anthocyanins (82%-100% decline) due to pH variations. However, specific phenolics, including p-hydroxybenzoic, protocatechuic, vanillic, p-coumaric, gallic and syringic acids, and coumarin esculetin, increased in concentration (10%-120%). Grape juice and wine samples showed comparable polyphenolic profile during all phases of digestion. Antioxidant activity persisted, and inhibition of angiotensin-I converting enzyme was improved after the digestion process, likely because of increased concentrations of listed phenolic acids and esculetin. Digested grape juice displayed comparable or superior bioactivity to red wine, indicating it as a promising source of accessible grape polyphenols for a broader audience. Nevertheless, Caco-2 cell model metabolization experiments revealed that only 3 of 42 analyzed compounds passed to the basolateral compartment, emphasizing the significant impact of digestion on polyphenol bioactivity, suggesting potential yet unmeasurable and overlooked implications for human health.

Polyphenolic profile and in vitro biological activity of Serbian orange (skin fermented white) wines.

Orange wines are made from white grapes, but with prolonged skin contact during fermentation. Available data on their composition and potential health benefits are limited, so polyphenolic profile (HPLC analysis) and in vitro biological activities (enzyme inhibition, antioxidant, and anti-inflammatory) of 24 Serbian orange wines were analyzed, including the correlation between determined composition and bioactivities. The wines displayed distinct polyphenolic profiles, enabling partial differentiation based on overall polyphenol content, including dominant components (catechin, gallic and caffeic acids), along with occasional occurrences of anthocyanins. However, no discernible distinctions were noted based on grape varieties, vintage, or producer. All twenty-four orange wines showed a reasonable inhibition of digestive enzymes and lipid peroxidation, twenty-one samples reduced ROS generation in the cell-based assay, but only two suppressed both PGE2 and TXA2 production in U937 cells, implicating possible functional food properties. No significant correlation between polyphenolic profile and determined biological activities was noticed.

Comprehensive study of phenolic profile and biochemical activity of monovarietal red and white wines from Fruska Gora region, Serbia.

This paper presents a comprehensive study of commercially available monovarietal red and white wines from Fruska Gora mountain in Serbia, conducted to characterize their chemical and biochemical properties. Namely, 72 wines of 18 different Vitis vinifera varieties, including international, domestic and autochthonous ones, were assessed. Phenolic profile of wines was determined qualitatively, using spectrophotometric methods, and quantitatively, using LC-MS/MS and HPLC-UV/VIS methods. Quantified phenolic compounds pointed out differences between red and white wines, where some compounds stood out as distinctive characteristic of analyzed varieties, while others served as chemotaxonomic markers of the Fruska Gora region. Antioxidant and inhibitory properties towards acetylcholinesterase were evaluated for all wines and their relation to phenolic composition was investigated. This study is valuable since chemical and biochemical properties of domestic and autochthonous Fruska Gora varieties are reported for the first time. Also, international varieties studied herein gave distinctive information about the terroir of Fruska Gora region, which is scarce in scientific literature.

Notifications and Health Consequences of Unauthorized Pharmaceuticals in Food Supplements.

Health concerns associated with the consumption of food supplements rise in parallel with the rise in the popularity and market availability of these products. In pursuit of data related to the unauthorized presence of pharmaceuticals in food supplements, the Rapid Alert System for Food and Feed (RASFF) database was searched for the 2011-2022 period. The most "popular" pharmaceuticals for the adulteration of food supplements were phosphodiesterase-5 inhibitors (235 records); anorexics and laxatives (76), including sibutramine and its active metabolite N-didesmethyl sibutramine, phenolphthalein and 2,4-dinitrophenol; stimulants, among which 1,3-dimethylamine (97), and synephrine (53) were the most numerous; nootropic drugs (24); anabolics and prohormones (16); and cannabinoid cannabidiol (14) (pending authorization as a novel food ingredient). Over 65% of notifications of interest were classified as serious risks, and over 80% of these were alert or border rejection notifications, mainly generated as a result of official control on the market. The alarming number of RASFF notifications should be considered a public health issue, demanding clear and targeted recommendation for action for the legislature and authorities. A harmonized nutrivigilance system should be considered as a tool to detect and scrutinize the adverse health effects of food supplements, along with measures to improve their safety, quality, and testing.

Health risk assessment of ethyl carbamate in fruit spirits.

Alcoholic beverages are multicomponent mixtures containing several carcinogenic compounds, including ethyl carbamate, which was evaluated in fruit spirits against the limit proposed by the International Alliance for Responsible Drinking (IARD) and then in terms of associated health. It was above the limit in 40% of the samples and statistically higher in stone fruit spirits. The MOE values below the limit were obtained for 49% and 98% of the samples at average and chronic heavy consumption by men, while for women the limit was reached by 19% and 96% of the samples, respectively. The lifetime cancer risk (LCR) approach based on the oral slope factor indicated risk concern for 96% of the samples at average consumption, regardless of gender, while LCR based on virtually safe dose indicated 25% and 69% of the samples posing a risk to women's and men's health, respectively. Preventive measures during spirit production and legal regulation of ethyl carbamate should be considered.

Health concerns associated to tropane alkaloids in maize food products in Serbia.

Following the implementation of the European regulation limiting the presence of tropane alkaloids in certain foods, a survey was conducted in Serbia on 103 maize products (grits, polenta and semolina) to determine atropine and scopolamine content using liquid chromatography with tandem-mass spectrometry analysis (LC-MS/MS). The probability of exceeding the Acute Reference Dose (ARfD; 0.016 µg/kg bw/day) of the sum of atropine and scopolamine by consuming these products was tested. Overall, across age categories - children, younger and older adolescents, and adults, the group ARfD was exceeded by 21.4%, 17.5%, 11.7% and 11.7% of the samples, with maximum exposure reaching as much as 19-, 13-, 9- and 9-fold the group ARfD, respectively. Nevertheless, polenta could be the most favorable dietary option (17.9% of positive samples, 7.7% resulting in excessive exposure in children, reaching a maximum of 1.4-fold the group ARfD). According to the reported findings, adverse health effects of tropane alkaloids cannot be ruled out. The Margin of Exposure, founded on a clinically significant acute effects dose established by FAO/WHO, ranged from 1194 to 2381 (mean) and from 28 to 56 (95th percentile) across age categories. These estimates should certainly draw the attention of food authorities and nutritionist, particularly in the case of highly sensitive populations with contraindications and high consumers of corn products, such as coeliac patients.

Benzoates and in situ formed benzene in food supplements and risk assessment.

Upward trend in the use of food supplements urged the assessment of their safety. Eighty-eight liquid herbal supplements collected in Novi Sad (Serbia) in 2018 (36 samples) and 2021 (52 samples) were analysed for the presence of benzoates and sorbates (HPLC-UV) and benzene (HS-GC/MS). Benzoic acid varied from 599 to 9253 mg/kg and sorbic acid between 185 and 1658 mg/kg. The acceptable daily intake of sorbic acid was not reached, but in case of benzoic acid, it was exceeded by 5.3% of the samples. The presence of benzene was confirmed in 41.2% of benzoate preserved supplements (0.9-51.7 µg/kg). Benzene exposure revealed no health concern: maximum hazard quotients ranged from 0.39% (toddlers) to 0.84% (adolescents); minimum margins of exposure were between 35,680 (adolescents) and 77,419 (toddlers); estimates of lifetime cancer risk did not reach one extra cancer case per 100 000 persons. However, measures to mitigate benzene presence in food should be considered.

Elemental Profile and Health Risk of Fruska Gora Wines.

The elemental composition of wine is influenced by endogenous sources and interventions from winemakers. The ICP-MS analysis of Fruska Gora wines (113) from vintages spanning across a decade (2011-2020), produced by 30 wineries and representing 18 autochthonous and international wine varieties, allowed a comprehensive insight into their elemental composition. Based on the mean concentrations of 23 investigated elements, B, Fe, and Mn, which were determined in mg per L of wine regardless of its colour or origin, were the most abundant. Red and white wines showed significant concentration differences in the case of B, Mn, and Sr (higher in red) as well as Be, Al, V, As, Mo, and Pb (higher in white). The elements of the highest toxicological concern were found in all (Pb and As) or almost all of the samples (Cd and Hg). Pb levels (maximum 47.1, 61.6, and 73.2 µg/L in red, rose, and white, respectively) were well below the legal limit. The applied risk assessment approaches (hazard quotient and index, margin of exposure) revealed no health concerns associated with consumption of Fruska Gora wines, except for a slightly increased lifetime cancer risk in the case of high wine consumption, and thus supported the promotion of Fruska Gora wines in the highly competitive international market.

Safety Assessment of Herbal Food Supplements: Elemental Profiling and Associated Risk.

Increased usage of herbal food supplements by both the paediatric and adult populations prompted a health risk assessment study. The elemental profiles of 52 supplements collected in Serbia were obtained using ICP-MS. The calculated contribution to permitted daily exposure was in compliance with the guidelines for elemental impurities: up to 46.8% for Pb (infants), 67.2% for Cu (toddlers), 6.8% for As (preschool children), and 8.0% for Hg (adolescents). Hazard quotients indicated high exposure of toddlers to Cu (100.9%) and Zn (112.7%), although by only one supplement, for which hazard indices ranged from 229 to 105% (aligning from toddlers to adolescents). Based on the margin of exposure, As and Pb were not of safety concern. Conversely, the proportion of supplements exceeding the acceptable level of lifetime cancer risk due to As exposure varied from 41.9% in adolescents to 54.3% in adults. Simultaneous use of multiple supplements over a prolonged period of time could further deepen health concerns.

Comparative Health Risk Assessment of Heated Tobacco Products versus Conventional Cigarettes.

BACKGROUND: Smoking is the largest preventable cause of death in the world with around eight million estimated premature deaths per year. In response to the harmful effects of conventional tobacco products the tobacco industry has launched a new type of products called Heated tobacco products (HTP) and e-cigarettes, which are considered safer for human health than conventional cigarettes. MATERIALS AND METHODS: The research was conducted by searching the scientific literature using platforms "Google scholar," "PubMed" and "Science Direct." To compare the non-carcinogenic and carcinogenic risk of HTP products and conventional cigarettes, margin of exposure and lifetime excess cancer risk were calculated using data obtained from a scientific literature search.HTP products have shown a reduced risk to human health compared to the conventional cigarettes, although they still contain compounds that can be dangerous to human health. There is not enough data obtained from independent studies that could safely indicate that these reduced amounts of toxic chemical entities in the composition of HTP do not induce any harmful effect. CONCLUSION: Further research is needed to determine the harmful effects of HTP aerosol, as well as to tighten the legislation that would limit the production, import and distribution of these products worldwide until their safety for human health is confirmed with a sufficient number of transparent and representative results obtained in independent scientific studies.

Phytochemistry and Antihyperglycemic Potential of Cistus salviifolius L., Cistaceae.

Cistus salviifolius has been previously reported as a traditional remedy for hyperglycemia. However, the plant has been scarcely investigated from scientific point of view. Thus, the aim was to examine the chemical composition and to evaluate its antioxidant and antihyperglycemic potential in vitro. Aqueous and ethanolic extracts were evaluated for total phenolic, tannin, and flavonoid content using spectrophotometric methods. Detailed chemical characterization was performed by high-performance liquid chromatography (HPLC-DAD). The volatile organic compounds (VOCs) profile was assessed by gas chromatography technique. The potential in diabetes treatment was evaluated through tests of free radicals neutralization, inhibition of lipid peroxidation process, and test of ferric ion reduction; activity in tests of inhibition of α-amylase, α-glucosidase and dipeptidyl peptidase-4 was also evaluated. High content of phenolics (majority being tannins) was detected; detailed HPLC analysis revealed high content of gallic acid, followed by rutin, chlorogenic and caffeic acids. The VOCs analysis determined sesquiterpene hydrocarbons and oxygenated sesquiterpenes as the main groups of compounds. The assays classified extracts as potent neutralizers of 2,2-diphenyl-1-picrylhydrazil and nitroso radicals formation and potent inhibitors of α-amylase and α-glucosidase. In conclusion, Cistus salviifolius represents a rich source of phenolics and essential oil with sesquiterpenes. The established results suggested its promising antioxidant and antihyperglycemic activities.

Design of experiments (DoE) to model phenolic compounds recovery from grape pomace using ultrasounds.

Grape processing by-products (particularly grape pomace) are known to contain high amounts of phenolic compounds. To improve the extraction of phenols from this by-product, it is necessary to develop a method and set and model optimal conditions for their extraction. By applying the design of experiments (DoE) approach, optimal experimental factors of Ultrasound-assisted extraction (USAE) were determined to obtain grape pomace extracts with a satisfactory yield of phenols anthocyanins, as well as extracts with high antioxidant capacity using reagents approved in the food industry. Initial method optimization covered two experimental factors: solvent concentration and the weight ratio of the sample and solvent using fixed USAE conditions from literature. For the final method optimization, the three investigated experimental factors were: pH value, the temperature of extraction, and extraction time. The optimal experimental conditions for the development of the method were 55% ethanol, sample/solvent ratio 1:40, pH 4.5, T 55 °C, and 30 min. Depending on the primary goal of the extraction process (the antioxidant activity, total phenolic content, content of individual phenols, or content of individual anthocyanins), these parameters can easily be modified to obtain the desired recovery. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05317-9.

Pesticide active substances in infant food in Serbia and risk assessment.

Risks posed by pesticide residues in infant food urge for protection of the most vulnerable part of our population. In the current study a total of 54 samples of infant food (juice and purée) were collected on the Serbian market. Liquid chromatography-tandem mass spectrometry with electrospray ionization method detected 18 out of 69 analyzed pesticide active substances in 55.6% of the samples, most frequently carbendazim and acetamiprid. Domestic products as opposed to the imported ones showed a substantially higher proportion of positive (85% vs 38%) and noncompliant (10% vs 0%) samples, a number of pesticides detected (15 vs 8), the proportion of the samples with multiple residues (85% vs 15%), the maximum number of residues in an individual sample (7 vs 2). Risk assessment was performed for the present pesticide active substances, which was estimated to remain below the level of concern for both acute and chronic adverse health effects.

Internet Marketing of Cardioprotective Dietary Supplements.

Objectives: Considering high prevalence of use of dietary supplements and their easy access on the internet, the aim of this research was to examine and assess the prevalence of the internet marketing of heart-protective supplements as the most popular supplements of today, and to investigate the quality and quantity of information that are available to consumers on the sale websites. Design: Three major search engines (Google, Yahoo, and Bing) and keywords "cardiovascular supportive supplements online buy" were used to identify websites that sell cardioprotective dietary supplements. Content of first 50 listed websites in each engine was evaluated for its compliance with regulatory acts while information about supplements' efficacy and safety was compared with the results of the latest scientific research. Results: Of a total of 150 listed websites, 89 selling supplements for the specific indication underwent further analysis. The most commonly registered cardioprotective dietary supplements on the internet were supplements based on omega-3 fatty acids (omega-3) (57 websites, 64.05%). Related to the websites selling omega-3 supplements, risk reduction claims were presented at 23 (40.35%), whereas structure or function claims were present at 50 (87.72%) analyzed websites, but followed with Food and Drug Administration disclaimer only on 68.00% of them. Information about adverse effects were rarely pointed out (1 website, 1.75%) unlike warnings, which were significantly more available to consumers (38, 66.67%). Conclusions: According to obtained results, most of the analyzed websites that sell omega-3 supplements did not contain all important medical information required by Dietary Supplement Health and Education Act. Since use of internet marketing is in expansion and since consumers have no access to relevant medical information about dietary supplements on the selling websites, there is a clear need for better quality control of websites and greater public awareness of these widely used products.

Biogenic amine and microbiological profile of Serbian dry fermented sausages of artisanal and industrial origin and associated health risk.

From the public health point of view, safety of dry fermented sausages is of concern due to possible presence of biogenic amines and some other food hazards. Taking into consideration that biogenic amines are chemical indicators of microbiological contamination, industrial and artisanal dry fermented sausages produced in Serbia were subjected to the microbiological and biogenic amine profiling. Microorganisms identified by MALDI-TOF MS included Cronobacter sakazakii and Listeria innocua, whereas the food pathogens were not detected. Biogenic amine content, determined by HPLC method in the range from 37.3 to 1186 mg/kg, was characterised as very low to low in 40% of the samples, moderate in 38%, high in 14% and very high in 8%. Risk assessment revealed that consumption of dry fermented sausages could cause histamine and tyramine intake up to 11.9% and 3.4% of threshold dose for healthy population, respectively. Adverse health effects would be rather unlikely for general population.

Health risk assessment for pediatric population associated with ethanol and selected residual solvents in herbal based products.

In this study, 48 herbal based products (41 for the pediatric population) were analyzed for the presence of ethanol and residual solvents. Ethanol was not detected in only 12% of the products designed for infants or toddlers aged under 2, and not quantified in only 5 of 14 'alcohol free' products. Actual content was higher than labeled in six out of 11 samples with specified ethanol quantity. WHO proposed requirement for ethanol content in products intended for use in children under the age of 6 (<0.5%) was not met by as many as 26 samples. Furthermore, calculated blood alcohol levels in children exceeded the relevant toxicological levels for nine samples following a single dose, and for one sample in case of accidental poisoning with the entire package. Regarding the residual solvents, acetone, 1-propanol and 1-butanol were not quantified, 2-propanol was found in two samples in low concentrations, whereas methanol intake via one of the samples exceeded the permitted level for children. The obtained results revealed a significant health concern for the pediatric population due to ethanol intake via herbal based products, calling for the establishment of strict guidelines for ethanol content and labeling.

Activity profiling of Serbian and some other European Merlot wines in inflammation and oxidation processes.

Merlot are worldwide recognized red wines. Several studies show that red wines have health benefits, mainly due to their phenolic constituents. This study evaluates twelve Serbian and other five European (French, Italian, Macedonian, Slovenian, Spanish) Merlot wines in respect of their phenolic composition and biological activity. The latter was evaluated through a set of in vitro experiments related to common benefits of moderate red wine consumption in the prevention of cardiovascular diseases. Among the examined phenolics, the most abundant acid in all samples was the gallic acid (14.3-58.3 mg/L), catechin (9.1-49.3 mg/L) was the dominant flavonoid, malvidin-3-O-glucoside (2.63-66.5 mg/L) leading anthocyanin, whereas resveratrol was found in a usual concentration (0.18-4.67 mg/L). Differences determined in phenolic profiles, mainly in content of quercetin, rutin and p-coumaric acid, leaded to separation of Serbian from foreing Merlot wines. Results of standard antioxidant assays (DPPH•, ABTS•+ and •NO scavenger capacity reducing power (FRAP), lipid peroxidation) revealed French Merlot as the most potent, but also pointed out some Serbian samples. The correlation between the content of dominant phenolics and antioxidant activity was sporadic, but samples with the highest overall phenolic content, generally had higher antioxidant potential. Concentration of wines and number of cells in ant-inflammatory assay were chosen to mimic in vivo conditions. So, the potency of examined wines to decrease the production of macrophage-derived PGE2 and TXA2 (up to 65.5 and 47.9%, respectively), could be considered as in vitro evidence of positve health effect. Regarding the phenolic content and anti-inflammatory contribution of the most abundant compounds, no correlation was witnessed. In general, this study showed interesting potential of Serbian Merlot wines, comparable to health-promoting effects of renewed Eurepean ones.

Plasma free amino acid profile in quiescent Inflammatory Bowel Disease patients orally administered with Mastiha (Pistacia lentiscus); a randomised clinical trial.

BACKGROUND: Natural products have been studied regarding their effectiveness on Inflammatory Bowel Disease (IBD). HYPOTHESIS/PURPOSE: To examine the effects of Mastiha (Pistacia lentiscus var. Chia) on clinical course and amino acid (AA) profile of patients in remission. STUDY DESIGN: This is a randomised, double-blind, placebo-controlled clinical trial. METHODS: Patients (n = 68) were randomly allocated to Mastiha (2.8  g/day) or placebo adjunct to stable medication. Free AAs were identified applying Gas Chromatography-Mass Spectrometry in plasma. Medical-dietary history, Inflammatory Bowel Disease Questionnaire, Harvey-Bradshaw Index, Partial Mayo Score, biochemical, faecal and blood inflammatory markers were assessed. Primary endpoint was the clinical relapse rate at 6 months. Secondary endpoints included variations in free AAs, inflammatory biomarkers and quality of life. Statistical significance was set at 0.05. RESULTS: Concerning AAs and biochemical data, alanine (p = 0.006), valine (p = 0.047), proline (p = 0.022), glutamine (p < 0.001) and tyrosine (p = 0.043) along with total cholesterol (p = 0.032) and LDL cholesterol (p = 0.045) increased only in placebo group compared with baseline and the change between the study groups was significantly different. Inflammatory markers had not a significantly different change between the two groups, even serum IL-6, faecal calprotectin and faecal lactoferrin increased only in the placebo group. Although Mastiha was not proven superior to placebo in remission rate (17.6% vs. 23.5%, p = 0.549), attenuation in increase of free AAs levels in verum group is reported. CONCLUSION: Mastiha inhibited an increase in plasma free AAs seen in patients with quiescent IBD. Since change of AAs is considered an early prognostic marker of disease activity, this indicates a potential role of Mastiha in remission maintenance.

Resveratrol supplementation improves metabolic control in rats with induced hyperlipidemia and type 2 diabetes.

Resveratrol was recognized as the major factor responsible for the beneficial properties of red wine. Several resveratrol-based dietary supplements are available, but their efficacy has not been sufficiently tested. This study was designed to examine the effect of resveratrol supplementation, using a commercially available product, on the metabolic status of experimental animals with induced hyperlipidemia or type 2 diabetes mellitus (T2DM). Hyperlipidemia was induced by feeding the rats a standard pellet diet supplemented with cholesterol. T2DM was induced by adding 10% fructose to drinking water and streptozotocin. Treatment with resveratrol-based supplement improved glycemic control in diabetic animals and significantly decreased serum low-density-lipoprotein (LDL) and triglyceride levels, concurrently increasing the high-density-lipoprotein (HDL) levels in animals with hyperlipidemia. Resveratrol-treated animals had improved tolerance to glucose loading. Supplementation did not induce alterations in parameters of liver and renal function. Findings indicate that commercial resveratrol supplement improves metabolic control in rats with induced hyperlipidemia and T2DM.

Antioxidative Efficacy of a Pistacia Lentiscus Supplement and Its Effect on the Plasma Amino Acid Profile in Inflammatory Bowel Disease: A Randomised, Double-Blind, Placebo-Controlled Trial.

Oxidative stress is present in patients with Inflammatory Bowel Disease (IBD), and natural supplements with antioxidant properties have been investigated as a non-pharmacological approach. The objective of the present study was to assess the effects of a natural Pistacia lentiscus (PL) supplement on oxidative stress biomarkers and to characterise the plasma-free amino acid (AA) profiles of patients with active IBD (Crohn's disease (CD) N = 40, ulcerative colitis (UC) N = 20). The activity was determined according to 5 ≤ Harvey Bradshaw Index ≤ 16 or 2 ≤ Partial Mayo Score ≤ 6. This is a randomised, double-blind, placebo-controlled clinical trial. IBD patients (N = 60) were randomly allocated to PL (2.8 g/day) or to placebo for 3 months being under no treatment (N = 21) or under stable medical treatment (mesalamine N = 24, azathioprine N = 14, and corticosteroids N = 23) that was either single medication (N = 22) or combined medication (N = 17). Plasma oxidised, low-density lipoprotein (oxLDL), total serum oxidisability, and serum uric acid were evaluated at baseline and follow-up. OxLDL/LDL and oxLDL/High-Density Lipoprotein (HDL) ratios were calculated. The plasma-free AA profile was determined by applying a gas chromatography/mass spectrometry analysis. oxLDL (p = 0.031), oxLDL/HDL (p = 0.020), and oxLDL/LDL (p = 0.005) decreased significantly in the intervention group. The mean change differed significantly in CD between groups for oxLDL/LDL (p = 0.01), and, in the total sample, both oxLDL/LDL (p = 0.015) and oxLDL/HDL (p = 0.044) differed significantly. Several changes were reported in AA levels. PL ameliorated a decrease in plasma-free AAs seen in patients with UC taking placebo. In conclusion, this intervention resulted in favourable changes in oxidative stress biomarkers in active IBD.