A Novel Colorimetric Nano Aptasensor for Ultrasensitive Detection of Aflatoxin B1 Based on the Exonuclease III-Assisted Signal Amplification Approach

The detection of aflatoxin B1 (AFB1) has recently garnered much attention on the issue of food safety. In this study, a novel and sensitive aptasensor towards AFB1 is proposed using an Exonuclease III (Exo III)-integrated signal amplification strategy. This reported sensing strategy is regulated by aptamer-functionalized nanobeads that can target AFB1; furthermore, complementary DNA (cDNA) strands can lock the immobilized aptamer strands, preventing the signal amplification function of Exo III in the absence of AFB1. The presence of AFB1 triggers the displacement of cDNA, which will then activate the Exo III-integrated signal amplification procedure, resulting in the generation of a guanine (G)-rich sequence to form a G-4/hemin DNAzyme, which can catalyze the substrate of ABTS to produce a green color. Using this method, a practical detection limit of 0.0032 ng/mL and a dynamic range of detection from 0.0032 to 50 ng/mL were obtained. Additionally, the practical application of the established sensing method for AFB1 in complex matrices was demonstrated through recovery experiments. The recovery rate and relative standard deviations (RSD) in three kinds of cereal samples ranged from 93.83% to 111.58%, and 0.82% to 7.20%, respectively, which were comparable with or better than previously reported methods.

Development and Validation of an LC-MS/MS Based Method for the Determination of Deoxynivalenol and Its Modified Forms in Maize

The Fusarium mycotoxin deoxynivalenol (DON) is a common contaminant of cereals and is often co-occurring with its modified forms DON-3-glucoside (D3G), 3-acetyl-DON (3ADON) or 15-acetyl-DON (15ADON). A stable-isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) based method for their determination in cereals was developed and validated for maize. Therefore, 13C-labelled D3G was enzymatically produced using 13C-DON and [13C6Glc]-sucrose and used as an internal standard (IS) for D3G, while uniformly 13C labelled IS was used for the other mycotoxins. Baseline separation was achieved for the critical peak pair DON/D3G, while 3ADON/15ADON could not be fully baseline separated after testing various reversed phase, fluorinated phase and chiral LC columns. After grinding, weighing and extracting the cereal samples, the raw extract was centrifuged and a mixture of the four 13C-labelled ISs was added directly in a microinsert vial. The subsequent analytical run took 7 min, followed by negative electrospray ionization and selected reaction monitoring on a triple quadrupole MS. Maize was used as a complex cereal model matrix for validation. The use of the IS corrected the occurring matrix effects efficiently from 76 to 98% for D3G, from 86 to 103% for DON, from 68 to 100% for 15ADON and from 63 to 96% for 3ADON.

Pesticides Contamination of Cereals and Legumes: Monitoring of Samples Marketed in Italy as a Contribution to Risk Assessment

The evaluation of cereal-based product contamination by pesticide residues is a topic of worldwide importance, and reliable analytical methods for official check analyses and monitoring studies are required for multi-residue analysis at trace levels. In this work, a validated multi-residual analytical method by gas-chromatography and tandem mass spectrometry coupled with a rapid QuEChERS procedure was used for the determination of 37 pesticides (pyrethroids, organophosphorus and organochlorine compounds) in 209 commercially available samples of cereals and 11 legumes, placed on the Italian market in 2018 and 2019, coming from different regions of Italy, eastern Europe, and some non-European countries. No pesticide traces were observed in the analyzed legume samples. A total of 18 cereal samples were found to be contaminated by at least one pesticide, with a concentration level higher than the corresponding quantification limit, but never exceeding the maximum level fixed in the European Regulations. This work is the first part of a surveillance study for pesticide control in food samples.

Occurrence of Ergot Alkaloids in Barley and Wheat from Algeria

The natural occurrence of six major ergot alkaloids, ergometrine, ergosine, ergotamine, ergocornine, ergokryptine and ergocristine, as well as their corresponding epimers, were investigated in 60 cereal samples (barley and wheat) from Algeria. Ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) and a QuEChERS extraction method were used for sample analysis. The results revealed that 12 out of 60 samples (20%) were contaminated with ergot alkaloids. Wheat was the most contaminated matrix, with an incidence of 26.7% (8 out of 30 samples). The concentration of total ergot alkaloids ranged from 17.8 to 53.9 µg/kg for barley and from 3.66 to 76.0 μg/kg for wheat samples. Ergosine, ergokryptine and ergocristine showed the highest incidences in wheat, while ergometrine was the most common ergot in barley.

ELISA and UPLC/FLD as Screening and Confirmatory Techniques for T-2/HT-2 Mycotoxin Determination in Cereals

T-2 and HT-2 toxins are secondary metabolites of various species of Fusarium. These molecules can have high potential toxic effects for human and animal health. In this work, ELISA and ultra performance liquid chromatography with fluorescence detection (UPLC/FLD) were implemented and validated as screening and confirmatory tests for the detection of these two toxins in cereal samples. The developed methods were tested by analyzing 100 samples of cereals by ELISA screening for reducing costs and analysis time and then using UPLC/FLD for confirmation purposes. Both methods met the performance criteria for sensitivity, linearity, selectivity, precision, and ruggedness, as reported in the European Decision No. 2002/657/EC and in Regulation (EC) No. 401/2006. The correlation between ELISA and UPLC/FLD approaches showed good results (r = 0.9056), confirming that these two techniques should be considered to be complementary in the official control activities of cereal and derived products.

Deep eutectic solvent based ultrasound assisted emulsification microextraction for preconcentration and voltammetric determination of aflatoxin B1 in cereal samples

A new and simple deep eutectic solvent based ultrasound-assisted emulsification microextraction (DES-UAEME) procedure has been developed for preconcentration and voltammetric determination of aflatoxin B1 (AFB1) in cereal products.

In Situ Monitoring of Sugar Content in Breakfast Cereals Using a Novel FT-NIR Spectrometer

This research demonstrates simultaneous predictions of individual and total sugars in breakfast cereals using a novel, handheld near-infrared (NIR) spectroscopic sensor. This miniaturized, battery-operated unit based on Fourier Transform (FT)-NIR was used to collect spectra from both ground and intact breakfast cereal samples, followed by real-time wireless data transfer to a commercial tablet for chemometric processing. A total of 164 breakfast cereal samples (60 store-bought and 104 provided by a snack food company) were tested. Reference analysis for the individual (sucrose, glucose, and fructose) and total sugar contents used high-performance liquid chromatography (HPLC). Chemometric prediction models were generated using partial least square regression (PLSR) by combining the HPLC reference analysis data and FT-NIR spectra, and associated calibration models were externally validated through an independent data set. These multivariate models showed excellent correlation (Rpre ≥ 0.93) and low standard error of prediction (SEP ≤ 2.4 g/100 g) between the predicted and the measured sugar values. Analysis results from the FT-NIR data, confirmed by the reference techniques, showed that eight store-bought cereal samples out of 60 (13%) were not compliant with the total sugar content declaration. The results suggest that the FT-NIR prototype can provide reliable analysis for the snack food manufacturers for on-site analysis.

Ergot and Ergot Alkaloids in Cereal Grains Intended for Animal Feeding Collected in Slovenia: Occurrence, Pattern and Correlations

This four-year study reports the occurrence of ergot alkaloids (EAs) in cereals intended for animal feeding collected in Slovenia. A total of 517 samples of cereals were analysed using liquid chromatography-tandem mass spectrometry for the presence of EAs. The sample set included wheat, rye, triticale, oat, spelt and barley. The study revealed that 17% of the analysed cereal samples were contaminated with at least one ergot alkaloid. EAs have two epimeric forms: -ine and -inine. The incidence rates of the -ine and -inine forms in the analysed samples were 16% and 15%, respectively. The highest contamination rates were observed in rye (54%), oat (50%) and spelt (30%), where the highest mean concentrations of total EAs were also determined (502 µg/kg, 594 µg/kg and 715 µg/kg, respectively). However, the highest concentrations of total EAs were found in wheat and rye (4217 µg/kg and 4114 µg/kg, respectively). The predominant EAs were ergometrine, ergosine and ergocristinine. The occurrence of six or more ergot alkaloids was observed in 49% of the positive samples. A weak correlation (p = 0.284) in the positive samples was found between the mass of sclerotia and the total concentrations of EAs using the Spearman correlation coefficient.