Chemical Characteristics of Croatian Traditional Istarski prsut (PDO) Produced from Two Different Pig Genotypes.

Chemical characteristics of raw and processed Istarski prsut (PDO) produced from two different pig genotypes were studied with special emphasis on amino and fatty acid composition and factors of lipid stability. Raw hams of Large White (LW)xLandrace (L), and (LWxL)xDuroc (D) pig genotypes were used in the study (20 hams of each genotype). All left raw hams from each carcass were processed in accordance with the PDO specification of Istarski prsut, and other half (the right ones) of LWxL)xD genotype were used for analyses of raw hams (fresh muscles). Istarski prsut was evaluated on the basis of the chemical parameters of the raw and matured lean ham. The process of dry curing significantly influenced the chemical properties of Istarski prsut. Despite the higher content of intramuscular fat and polyunsaturated fatty acids, the fat of (LWxL)xD ham was much more resistant to hydrolysis and oxidation, suggesting that fatty acid profile and other factors, also play a significant role. Significant differences between pig genotypes in the amino acid and fatty acid profiles were found. The analyzed Istarski prsut may be distinguished by prints of multivariate chemometric statistical analysis, based on their amino acid and fatty acid compositions.

VERIFICATION OF AUTHENTICITY OF GINKGO BILOBA L. LEAF EXTRACT AND ITS PRODUCTS PRESENT ON THE CROATIAN MARKET BY ANALYSIS OF QUANTITY AND RATIO OF GINKGO FLAVONE GLYCOSIDES (QUERCETIN, KAEMPFEROL AND ISORHAMNETIN) TO TERPENE TRILACTONES TO THE EFFECT OF UNMASKING COUNTERFEIT DRUGS ENDANGERING PATIENT HEALTH.

Ginkgo biloba L. is the eldest plant growing on the Earth; preparations made of its leaves and seeds represent an integral part of the Chinese medicine for over a millennium. The plant species was first discovered by Linnaeus in 1771, its name thereby originating from the Latin words bis (two) and lobus (lobe), which duly illustrate the specific shape of its leaf. Contemporary Ginkgo biloba L. plant based pharmaceuticals mostly comprise extracts recovered from leaves harvested during fall, when the concentration of active components reaches its peak. Recent investigations have managed to establish the chemical composition of the plant leaf, together with the mechanisms underlying its beneficial effects on rheological profile of the blood and acceleration of its flow. High price of these preparations and their vast popularity have soon become an incentive for counterfeiting Ginkgo biloba L. extracts and the release of bogus drugs comprising cheaper extracts coming from other plants. Namely, modern Ginkgo biloba L.-based medicinal products and food supplements comprise extracts recovered from the plant leaf that get to be standardized according to its key pharmacological active components, most often flavone glycosides (represented in the share of 22%-27%) and terpene trilactones (represented in the share of 6%-7%). The flavonoids that predominate such preparations and are most relevant from the pharmacological standpoint are quercetin, kaempferol and isorhamnetin, their total amount and mutual ratios, thereby being an unquestionable indicator of the extract authenticity. Therefore, most of the analyses aiming at verifying the authenticity of a given Ginkgo biloba L.-based product boil down to the analysis of these parameters. Counterfeiting involves partial or full replacement of the Ginkgo biloba L. extract (GBE) with a cheaper plant extract of a similar composition, the latter occasionally being enriched with an additional amount of flavonoids, most often quercetin, not originating from the Ginkgo biloba L. plant. The aim of this study was to verify the authenticity and quality of Ginkgo biloba L.-based products circulating on the Croatian market. To that effect, 10 samples of products produced by various manufacturers were analyzed in a certified laboratory. The parameters based on which the authenticity of the preparations was assessed were the shares of aglycones of typical ginkgo flavone glycosides, that is to say, quercetin, kaempferol and isorhamnetin, and mutual ratios of the established quantities of quercetin to kaempferol as the key clues to unmasking Ginkgo extracts counterfeiting. The amount of ginkgo flavone glycosides was established using high performance liquid chromatography. The analysis proved 80% of the samples analyzed to be conformant to the label statements as regards the total amount of flavone glycosides and their mutual ratios. In 20% of the samples, the ratio of quercetin to kaempferol deviated from normal values; on top of that, the presence of the phytoestrogen genistein, one of the components typically comprised by the Sophora japonica L. plant, was also proven, documenting counterfeiting of the GBE and its replacement by the Sophora japonica L. extracts in the samples under consideration. Due to the untrue label statements descriptive of these products, the information on the presence of pharmacologically active genistein was neglected to be mentioned despite its unfavorable health impact that can be expected in some consumer groups. The results of this study indicated the frequency of counterfeiting the Ginkgo biloba L.-based products found on the Croatian market to be deemed substantial. Therefore, a more rigorous and more thorough control of these products and sanctioning of irresponsible manufacturers and distributers is proposed, so as to contribute to a higher market representation of high-quality products, as well as to avoid health risks and downsize the rate of their counterfeiting.

Heavy metals and metalloid content in vegetables and soil collected from the gardens of Zagreb, Croatia.

Aim of this study was to determine concentration of Pb, Cd, As and Hg in green leafy vegetables and soil in the urban area of Zagreb, Croatia and to determine if there is a connection between the contamination of soil and vegetables. Green leafy vegetables and soil samples were taken from the gardens located in the outskirts of the city. Concentrations of Pb, Cd, As and Hg were determined by atomic absorption spectrometry; showing that average concentrations of metals and metalloids in vegetables and in soil, regardless of the location of sampling were below the maximum allowed concentration (MAC). The analysis determined that metal concentrations in only nine vegetable samples (9%) were above maximum allowed values prescribed by national and European legislation (three with higher concentrations of Pb, one with a higher concentration of Cd and five with higher concentrations of Hg). Concentrations of contaminants present in the analysed samples, in general, are lower than the ones published in similar studies. The final distribution and concentration of contaminants in vegetables of Zagreb, besides industry and traffic, is affected by the dominant wind direction.

Development of a Pressurized Green Liquid Extraction Procedure to Recover Antioxidant Bioactive Compounds from Strawberry Tree Fruit (Arbutus unedo L.).

Strawberry tree fruits (Arbutus unedo L.) are a natural source of valuable biologically active compounds. Therefore, the aim of this work was to develop a green extraction approach using pressurized liquid extraction (PLE) to provide the highest yield of bioactive compounds. Different extraction solvents (distilled water, 50% and 96% ethanol) and different PLE parameters were investigated: temperature (40, 80, and 120 °C), static extraction time (5 and 10 min), and number of cycles (1 and 2). Total phenolic contents (TPC), hydroxycinnamic acids (HCA), flavonols (FL), total flavonoids (TF), condensed tannins (CT), and antioxidant capacity (DPPH and FRAP) were determined in PLE extracts. Solvent type, temperature, static extraction time, and number of cycles had a statistically significant effect on all bioactive compounds and antioxidant capacity (p ≤ 0.05). All bioactive compounds were positively correlated with temperature, except for TPC and DPPH. For all polyphenols studied, the optimal PLE parameters were a temperature of 120 °C, a static extraction time of 10 min and 2 cycles. The best solvent for most bioactive compounds was 96% ethanol, except for TPC, for which 50% ethanol was better. This study suggests that PLE is a promising tool to intensify the extraction of bioactive compounds from strawberry tree fruits.

Assessment of Metal Intake by Selected Food Supplements Based on Beehive Products.

The aim of this study was to determine the quantity of particular toxic metals (Pb, Cd, As, Hg) and micronutrients (Cr, Fe, Co, Ni, Cu, Zn, Se) in the recommended daily dose of 51 food supplements based on beehive products. Samples taken from the Croatian market were submitted for the identification/quantification of studied metals and micronutrients. It was carried out by means of inductively coupled plasma mass spectrometry (ICP-MS). Eleven samples (21.57%) showed an increased concentration of total arsenic, three samples (5.88%) contained an increased concentration of total iron, and eight samples (15.68%) had an increased concentration of total nickel. Three samples (5.88%) contained an increased concentration of zinc, while one sample (1.96%) contained an increased concentration of selenium. Increased levels of certain toxic metals and micronutrients do not pose a danger to human health because the amount identified was less than what can cause toxic effects in humans. All other analysed metals and micronutrients fell within the defined literature values. Despite certain increases in particular parameters, all samples met the established toxicity criteria. This study evidenced their safety if consumed in the recommended daily dose.

Unauthorized Food Manipulation as a Criminal Offense: Food Authenticity, Legal Frameworks, Analytical Tools and Cases.

Food fraud is a criminal intent motivated by economic gain to adulterate or misrepresent food ingredients and packaging. The development of a reliable food supply system is at great risk under globalization, but Food Business Operators (FBOs) have a legal obligation to implement and maintain food traceability and quality at all stages of food production, processing, and distribution. Incidents of food fraud have a strong negative impact on consumer confidence in the food industry. Therefore, local and international regulatory mechanisms are established to prevent or mitigate food fraud. This review brings new perspectives linking EU and US legislation, as well as new definitions and descriptions of the criminal aspect of food fraud incidents. It also describes certain new insights into the application of state-of-the-art methods and techniques that provide valuable tools for geographic, botanical, or other chemical markers of food authenticity. The review also provides an overview of the most common cases of food fraud worldwide from 2010 to 2020. Further research is needed to support the development of predictive models for innovative approaches to adulteration, especially when some valuable nutrients are replaced by toxic ingredients. A possible solution to minimize food fraud incidents is to increase the level of risk-based inspections, establish more productive monitoring and implementation of food protection systems in the supply chain, and implement better ingredient control and certification. National and international (e.g., regional) police offices for food fraud should be introduced, possessing knowledge and skills in food, food safety, food processing, and food products, as initial positive results have emerged in some countries.

Facts about phthalate toxicity in humans and their occurrence in alcoholic beverages.

Phthalates are esters of phthalic acid and aliphatic alcohol added to plastic to improve its softness, flexibility, and extensibility. They easily migrate from plastic products into the environment because of their physical and chemical properties. This review summarises their characteristics, distribution in the environment, monitoring, use, toxic effects on human health, regulatory limits in different matrices and products, and tolerable daily intake. The studies we have reviewed suggest that phthalates have a potential to affect reproduction and development in humans. Due to the inconsistent data, further studies are needed and, in the meantime, precautionary policies must be implemented. Here we draw attention to the methods of determining phthalate levels in alcoholic beverages and reported levels in plum spirits produced in Croatia. Legally produced and moderately consumed plum spirits do not seem to increase the risk of phthalate toxicity for human health. We conclude with recommendations for the effective monitoring of phthalate exposure in humans and for the implementation of alternative materials in alcohol production.

PRESENCE OF CITRININ IN GRAINS AND ITS POSSIBLE HEALTH EFFECTS.

BACKGROUND: Citrinin is a mycotoxin produced by several species of the genera Aspergillus, Penicillium and Monascus and it occurs mainly in stored grain. Citrinin is generally formed after harvest and occurs mainly in stored grains, it also occurs in other plant products. Often, the co-occurrence with other mycotoxins is observed, especially ochratoxin A, which is usually associated with endemic nephropathy. At the European Union level, systematic monitoring of Citrinin in grains began with the aim of determining its highest permissible amount in food. Thus, far the systematic monitoring of the above mentioned mycotoxin in Croatia is yet to begin. MATERIALS AND METHODS: The main goal of this study was to determine the presence of Citrinin in grains sampled in the area of Medimurje, Osijek-Baranja, Vukovar-Srijem and Brod-Posavina County. For the purpose of identification and quantification of citrinin, high performance liquid chromatograph (HPLC) with fluorescence was used (Calibration curve k > 0.999; Intra assay CV = 2.1%; Inter assay CV = 4.3%; LOQ < 1 µg/kg). RESULTS: From the area of Medimurje County, 10 samples of corn and 10 samples of wheat were analyzed. None of the samples contained Citrinin (<1 µg/kg). From the area of Osijek-Baranja and Vukovar-Srijem County, 15 samples from each County were analyzed. The mean value for the samples of Osijek-Baranja County was 19.63 µg/kg (median=15.8 µg/kg), while for Vukovar-Srijem County the mean value of citrinin was 14,6 µg/kg (median=1.23 µg/kg). From 5 analyzed samples from Brod-Posavina County, one of the samples contained citrinin in the amount of 23.8 µg/kg, while the registered amounts in the other samples were <1 µg/kg. CONCLUSION: The results show that grains from several Counties contain certain amounts of Citrinin possibly indicating a significant intake of citrinin in humans. It must be stated that grains and grain-based products are the basis of everyday diet of all age groups, especially small children, where higher intake of citrinin can occur. Consequently, we emphasize the need for systematic analysis of larger amount of samples, from both large grains and small grains, especially in the area of Brod-Posavina County, in order to obtain more realistic notion of citrinin contamination of grains and to asses the health risk in humans.

NITRATE IN LEAFY GREEN VEGETABLES AND ESTIMATED INTAKE.

BACKGROUND: Vegetarian diets are rich in vegetables. Green leafy vegetables are foods that contain considerable amounts of nitrate, which can have both positive and negative effects on the human body. Their potential carcinogenicity and toxicity have been proven, particularly after the reduction of nitrate to nitrite itself or just serving as a reactant with amines and/or amides in the formation of N-nitroso compounds -N-nitrosamines and other nitrogen compounds which may have high levels of nitrate. The aim of this study was to determine whether there is a significant difference, considering the location and seasonal sampling period, in the level of nitrate in certain types of green vegetables, all in order to be able to assess their intake, and possible impact on human health, especially knowing that exposure to nitrate can be potentially higher for vegetarian population group. MATERIALS AND METHODS: For this purpose, the sampling of 200 different leafy green vegetables was conducted, all of which could be found in free sale in the Republic of Croatia. The sampling was conducted during two seasonal periods - the spring and autumn period. In the springtime, lettuce (sem), spinach (pinacho), kale (kale), chard (mangel) and cabbage (brassica) were sampled, and in autumn lettuce, spinach, kale, chard and arugula. Samples were analyzed using high performance liquid chromatography (HPLC) with UV detection. RESULTS: The results from the spring sampling phase were in the range of 603 mg/kg for cabbage - 972 mg/kg for chard, and for autumn phase of 1.024 mg/kg for chard to 4.354 mg/kg for the arugula. The results showed that there were significant differences (p <0.05) for most of the samples analyzed, considering the sampling locations and time period. CONCLUSION: The results indicate that the analyzed vegetables contain significant amounts of nitrate in their composition, which represents relatively significant, but still acceptable intake into the human body.