Target quantification and semi-target screening of undesirable substances in pear juices using ultra-high-performance liquid chromatography-quadrupole orbitrap mass spectrometry

Fruit juices are common products in modern diets due to the supply of vegetal nutrients combined with its tastiness. Nevertheless, potential contaminants, such as mycotoxins and pesticides, can be present in commercial products due to a potential carry-over. Therefore, the aim of this study was to investigate for the first time the presence of 14 Fusarium mycotoxins using a quick, easy, cheap, effective, rugged, and safe (QuEChERS)-based extraction followed by an ultra-high-performance liquid chromatography-quadrupole Orbitrap high-resolution mass spectrometry in 21 pear juice samples from Italian markets. Up to nine different mycotoxins were detected, particularly an extensive presence of zearalenone (67%, n = 21, mean value = 0.88 ng/mL). Emerging Fusarium mycotoxins enniatins B, B1, A, and A1 were also detected. Additionally, 77 pesticide residues were tentatively identified through a retrospective analysis based on a mass spectral library. The prevalent presence of some non-approved pesticides, such as ethoxyquin (64%, n = 21) and triazophos (55%, n = 21), must be highlighted. The results obtained indicate an extensive contamination of marketed pear juice with undesirable compounds, and they should be taken into consideration when performing risk assessment studies

Ultra-high-performance liquid chromatography coupled with quadrupole Orbitrap high-resolution mass spectrometry for multi-residue analysis of mycotoxins and pesticides in botanical nutraceuticals

Cannabidiol (CBD) food supplements made of Cannabis sativa L. extracts have quickly become popular products due to their health-promoting effects. However, potential contaminants, such as mycotoxins and pesticides, can be coextracted during the manufacturing process and placed into the final product. Accordingly, a novel methodology using ultra-high-performance liquid chromatography coupled with quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was developed to quantify 16 mycotoxins produced by major C. sativa fungi, followed by a post-target screening of 283 pesticides based on a comprehensive spectral library. The validated procedure was applied to ten CBD-based products. Up to six different Fusarium mycotoxins were found in seven samples, the most prevalent being zearalenone (60%) and enniatin B1 (30%), both found at a maximum level of 11.6 ng/g. Co-occurrence was observed in four samples, including one with enniatin B1, enniatin A and enniatin A1. On the other hand, 46 different pesticides were detected after retrospective analysis. Ethoxyquin (50%), piperonyl butoxide (40%), simazine (30%) and cyanazine (30%) were the major residues found. These results highlight the necessity of monitoring contaminants in food supplements in order to ensure a safe consumption, even more considering the increase trend in their use. Furthermore, the developed procedure is proposed as a powerful analytical tool to evaluate the potential mycotoxin profile of these particular products

Simultaneous determination of afb1 and afm1 in milk samples by ultra high performance liquid chromatography coupled to quadrupole orbitrap mass spectrometry

Milk is the world’s most consumed beverage, not counting water. Even though investigations on milk aflatoxin (AF) M1 contamination are regularly conducted, there is limited information on the contamination of milk with its parent compound, AFB1. Hence, the aim of this study was to develop a quick, easy, cheap, effective, rugged, and safe (QuEChERS)-based method for the simultaneous analysis of AFB1 and AFM1 in milk, using ultrahigh performance liquid chromatography coupled with high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). The recoveries were in a range of 75–96% at 0.005, 0.01, and 0.05 µg/L spiking levels, with repeatability and reproducibility results expressed as relative standard deviations (RSDs) lower than 7% and 16%, respectively. The limits of detection (LODs) and quantification (LOQs) were 0.001 and 0.002 µg/L for AFM1 and AFB1, respectively. The LODs and LOQs that were obtained showed the suitability of the developed method for the determination of trace amounts of the selected mycotoxins in milk samples, and were up to ten times lower than those that had been reported in previous works using triple quadrupole mass analyzers. The matrix effect was evaluated and matrix-matched calibrations were used for quantification. The validated method was applied to 40 Italian milk samples. Neither AFB1 nor AFM1 were found above the LOD in any of the analyzed samples

Occurrence and exposure assessment of mycotoxins in ready-to-eat tree nut products through ultra-high performance liquid chromatography coupled with high resolution q-orbitrap mass spectrometry

Tree nuts have become popular snacks due to their attributed benefits in the health state. Nevertheless, their susceptibility to fungal contamination lead to the occurrence of potentially dangerous mycotoxins. Hence, the aim of this work was to evaluate the presence of mycotoxins in ready-to-eat almonds, walnuts, and pistachios from Italian markets. The most relevant mycotoxin found in almonds was α-zearalanol in 18% of samples (n = 17) ranging from 3.70 to 4.54 µg/kg. Walnut samples showed frequent contamination with alternariol, present in 53% of samples (n = 22) at levels from 0.29 to 1.65 µg/kg. Pistachios (n = 15) were the most contaminated commodity, with β-zearalenol as the most prevalent toxin present in 59% of samples ranging from 0.96 to 8.60 µg/kg. In the worst-case scenario, the exposure to zearalenone-derived forms accounted for 15.6% of the tolerable daily intake, whereas it meant 12.4% and 21.2% of the threshold of toxicological concern for alternariol and alternariol monomethyl-ether, respectively. The results highlighted the extensive presence of Alternaria toxins and zearalenone-derived forms, scarcely studied in ready-to-eat tree nut products, highlighting the necessity to include these mycotoxins in analytical methods to perform more realistic risk assessments

Human biomonitoring of t-2 toxin, t-2 toxin-3-glucoside and their metabolites in urine through high-resolution mass spectrometry

The metabolic profile of T-2 toxin (T-2) and its modified form T-2-3-glucoside (T-2-3-Glc) remain unexplored in human samples. Therefore, the present study aimed to investigate the presence of T-2, T-2-3-Glc and their respective major metabolites in human urine samples (n = 300) collected in South Italy through an ultra-high performance liquid chromatography (UHPLC) coupled to Q-Orbitrap-HRMS methodology. T-2 was quantified in 21% of samples at a mean concentration of 1.34 ng/mg Crea (range: 0.22–6.54 ng/mg Crea). Almost all the major T-2 metabolites previously characterized in vitro were tentatively found, remarking the occurrence of 3′-OH-T-2 (99.7%), T-2 triol (56%) and HT-2 (30%). Regarding T-2-3-Glc, a low prevalence of the parent mycotoxin (1%) and its metabolites were observed, with HT-2-3-Glc (17%) being the most prevalent compound, although hydroxylated products were also detected. Attending to the large number of testing positive for T-2 or its metabolites, this study found a frequent exposure in Italian population

Colon bioaccessibility under in vitro gastrointestinal digestion of different coffee brews chemically profiled through UHPLC-Q-orbitrap HRMS

Coffee represents one of the most traditionally consumed beverages worldwide, containing a broad range of human health–related compounds. According to previous studies, regular coffee consumption may display protective effects against colorectal cancer and other chronic diseases. The main goal of this research was to evaluate the bioaccessibility of phenolic content and variation in antioxidant capacity of three different types of coffee brews after simulated gastrointestinal digestion. This would allow to elucidate how antioxidant compounds present in coffee may exert their effect on the human body, especially in the colonic stage. Moreover, the content of bioactive compounds namely chlorogenic acids (CGAs, n = 11) and caffeine was also assessed throughout ultra-high-performance liquid chromatography followed by high-resolution Orbitrap mass spectrometry (UHPLC-Q-Orbitrap HRMS). The three main isomers of caffeoylquinic acid constituted the highest fraction of CGAs present in the samples, accounting for 66.0% to 70.9% of total CGAs. The bioaccessibility of coffee polyphenols significantly increased in digested samples from 45.9% to 62.9% at the end of the colonic passage, compared to the non-digested samples. These results point to the colonic stage as the major biological site of action of the active antioxidant coffee compounds

Target analysis and retrospective screening of mycotoxins and pharmacologically active substances in milk using an ultra-high-performance liquid chromatography/high-resolution mass spectrometry approach

Milk is a nutritious food suitable for infants and adults, and it plays an important role in the human diet. However, it may also be a vehicle for food contaminants. In this report, we developed a method using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-Q-Exactive Orbitrap HRMS; Thermo Fisher Scientific, Waltham, MA) for simultaneous identification of target pharmacologically active substances and mycotoxins in milk. We also used the Q-Orbitrap operating in full scan mode to identify other possible drugs and microbial metabolites that occurred in samples. Fifty-six commercially available milk samples from the Italian market were analyzed. Investigated analytes were extracted using a QuEChERS (quick, easy, cheap, effective, rugged, and safe) approach. Method detection and quantification limits and performance criteria set by European regulations were fulfilled. Pharmacologically active substances were detected in 49% of samples (range 0.007–4.53 ng/mL), including nontarget mycotoxins. Retrospective analysis allowed us to identify other antibiotics and pharmacologically active substances, as well as nonregulated fungal/bacterial metabolites at a relatively high incidence. From the obtained values, the need for continuous monitoring of contaminants in the milk production chain is clear. This is the first study to assess the presence of pharmacologically active substances, mycotoxins, and other microbial metabolites in Italian milk samples using the UHPLC-Q-Orbitrap HRMS system

Biomonitoring of enniatin B1 and its phase i metabolites in human urine: First large-scale study

Enniatins (Enns) are mycotoxins produced by Fusarium spp. which are a fungus widely spread throughout cereals and cereal-based products. Among all the identified enniatins, Enn B1 stands as one of the most prevalent analogues in cereals in Europe. Hence, the aim of this study was to evaluate for the first time the presence of Enn B1 and its phase I metabolites in 300 human urine samples using an ultrahigh-performance liquid chromatography high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) methodology. Enn B1 was detected in 94.3% of samples ranging from 0.007 to 0.429 ng/mL (mean value: 0.065 ng/mL). In accordance with previous in vitro and in vivo analysis, hydroxylated metabolites (78.0% samples) and carbonylated metabolites (66.0% samples) were tentatively identified as the major products. Results from this biomonitoring study point to a frequent intake of Enn B1 in the studied population, suggesting that in-depth toxicological studies are needed in order to understand the potential effects in humans