In this study, green extraction methods—high voltage electrical discharges (HVED), pulsed electric field (PEF), and ultrasound-assisted extraction (UAE)—were compared in terms of extraction yield of total and individual polyphenolic compounds, as well as the antioxidant capacity of blueberry pomace extracts. All extractions were performed with methanol- and ethanol-based solvents. The highest total polyphenols content (TPC) (10.52 mg of gallic acid equivalent (GAE) per g of dry weight (dw)) and antioxidant activity (AA) (0.83 mmol TE/g dw) were obtained by PEF-assisted extraction in the ethanol-based solvent after 100 pulses and 20 kV/cm, which corresponds to an energy input of 41.03 kJ/kg. A total of eighteen individual polyphenols were identified in all investigated blueberry pomace extracts by high-performance liquid chromatography with the diode-array detector (HPLC-DAD) and liquid chromatography electrospray ionization tandem mass spectrometric (LC-(HESI)-MS/MS). The highest anthocyanin (1757.32 µg/g of dw) and flavanol (297.86 µg/g of dw) yields were obtained in the methanol-based solvent, while the highest phenolic acid (625.47 µg/g of dw) and flavonol (157.54 µg/g of dw) yields were obtained in the ethanol-based solvent by PEF-assisted extraction at the energy input of 41.03 kJ/kg. These results indicated that PEF is a promising green extraction method which can improve the blueberry pomace’s polyphenol extraction yield.

Cereals are still one of the most important food and feed sources, thus determining cereal’s safety, i.e., compliance with legislation, is extremely important. As systematic investigations of nowadays unavoidable secondary fungal metabolites and other common legally regulated contaminants occurrence in Croatian cereals are still lacking, this research aims to monitor the contamination levels of nation-wide crops by mycotoxins, pesticide residues, and heavy metals by employing UHPLC-MS/MS, GC-MS/MS, and atomic absorption spectrometer (AAS) validated analytical methods. The most common secondary fungal metabolites were found to be Fusarium mycotoxins, with DON being the most occurring present in 73.7% of the samples. At least one pesticide residue was found in 331.8% of the samples, and Hg and Cd were the most occurring heavy metals. A total of 8.5% of the samples was non-compliant to the European Union (EU) legislation for food regarding the found mycotoxins concentrations, 4.5% regarding pesticide residues and none regarding heavy metals. The unusual presence of certain pesticide residue and heavy metal indicates the importance of systematic control of the contaminant presence, in order to gather enough occurrence data for proper risk assessment that these contaminants represent for the consumer’s health.

Mycotoxins are secondary metabolites produced by toxigenic fungi in crops worldwide. In (micro)organisms such as plants, fungi, bacteria, or animals they may be further metabolised and modified, but this is also true for food processing, which may lead to a wide range of masked mycotoxin forms. These often remain undetected by analytical methods and are the culprits for underestimates in risk assessments. Furthermore, once ingested, modified mycotoxins can convert back to their parent forms. This concern has raised the need for analytical methods that can detect and quantify modified mycotoxins as essential for accurate risk assessment. The promising answer is liquid chromatography-mass spectrometry. New masked mycotoxin forms are now successfully detected by iontrap, time-of-flight, or high-resolution orbitrap mass spectrometers. However, the toxicological relevance of modified mycotoxins has not been fully clarified.

A total of 209 samples of various cereal crops (maize, wheat, barley, rye and oats) grown in Croatian fields during 2016 and 2017 were collected to analyze and determine the occurrence and co-occurrence of EU regulated mycotoxins in cereals (AFB1, AFB2, AFG1, AFG2, DON, FB1, FB2, ZEA, T-2, HT-2 and OTA). The analysis, performed by a validated confirmatory LC-MS/MS method based on a dilute and shoot principle, highlighted Fusarium mycotoxins as the main contaminants, often co-occurring in samples from both years (50.0% in 2016 and 33.7% in 2017). DON was found to be the most frequent mycotoxin, present in 72.5% of the 2016 samples and 32.6% of the 2017 samples, while maize proved to be the most contaminated cereal type of both years with FUM as the most abundant mycotoxins, with an average concentration of 1180 µg/kg. Moderate temperatures with periods of high humidity favored the accumulation of DON in wheat samples instead of other Fusarium mycotoxins, while similar conditions favored maize contamination with FUM. A total of 8.3% of all the 2016 harvest samples and 7.9% of the 2017 harvest samples were assessed as non-compliant, containing mycotoxins in concentrations higher than the levels set by the EU legislation for food.

Micro matrix solid phase dispersion (micro-MSPD) was optimized by response surface methodology for the extraction of polyphenols from the peel of twelve traditional and eight commercial apple varieties grown in Croatia. The optimized micro-MSPD procedure includes the use of 0.2 g of sample, 0.8 g of dispersant, a 57% solution of methanol in water as the solvent and 5 mL of extract volume. The total polyphenolic index (TPI) and antioxidant activity (AA) were measured by spectrophotometric assays. Eighteen polyphenolic compounds were identified in all investigated apples by HPLC-DAD and LC-(ESI)-MS. The peel of traditional apple varieties had higher contents of all investigated polyphenols. Calculated relative contribution of polyphenol groups indicated non-flavonoids (28.6%) and flavanols (46.2%) as the major contributors to the total polyphenolic content in traditional and commercial apple varieties, respectively. The most abundant polyphenol in traditional apple peel was chlorogenic acid, procyanidin B2 and epicatechin (1143 ± 755 µg/g dw, 954 ± 343 µg/g dw and 560 ± 362 µg/g dw, respectively). The peel of varieties ‘Apistar’, ‘Bobovac’ and ‘Božićnica’ could be highlighted as an important source of polyphenols.

Penicillium expansum is a necrotrophic plant pathogen among the most ubiquitous fungi disseminated worldwide. It causes blue mould rot in apples during storage, transport and sale, threatening human health by secreting patulin, a toxic secondary metabolite that contaminates apples and apple-derived products. Nevertheless, there is still a lack of sufficient data regarding the resistance of different apple cultivars to P. expansum, especially ancient ones, which showed to possess certain resistance to plant diseases. In this work, we investigated the polyphenol profile of 12 traditional and 8 conventional apple cultivar and their resistance to P. expansum CBS 325.48. Eight polyphenolic compounds were detected; the most prominent were catechin, epicatechin and gallic acid. The highest content of catechin was detected in ‘Apistar’—91.26 mg/100 g of fresh weight (FW), epicatechin in ‘Bobovac’—67.00 mg/100 g of FW, and gallic acid in ‘Bobovac’ and ‘Kraljevčica’—8.35 and 7.40 mg/100 g of FW, respectively. The highest content of patulin was detected in ‘Kraljevčica’ followed by ‘Apistar’—1687 and 1435 µg/kg, respectively. In apple cultivars ‘Brčko’, ‘Adamčica’ and ‘Idared’, patulin was not detected. Furthermore, the patulin content was positively correlated with gallic acid (r = 0.4226; p = 0.002), catechin (r = 0.3717; p = 0.008) and epicatechin (r = 0.3305; p = 0.019). This fact indicates that higher contents of gallic acid, catechin and epicatechin negatively affected and boost patulin concentration in examined apple cultivars. This can be related to the prooxidant activity of polyphenolic compounds and sensitivity of P. expansum to the disturbance of oxidative status.