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The aim of this study was to estimate the aflatoxin M1 (AFM1) contamination in raw milk and processed milk (pasteurized or UHT) collected from two regions in Italy (Puglia and Basilicata) during a 12-year period: 2012–2023. A total of 1017 milk samples were analyzed first proceeding with screening analysis by enzyme-linked immunosorbent assay (ELISA), and suspected non-compliant samples (AFM1 concentration higher than 0.042 µg/kg) were then analyzed by high performance liquid chromatographic with fluorimetric detection (HPLC/FLD) confirmation method. AFM1 concentration ≥ 0.005 µg/kg (ELISA limit of quantitation) was detected in 553 of the 1017 milk samples (54.4%). AFM1 levels exceeding the European Union maximum limit (ML) of 0.050 µg/kg were detected in 70 samples, 49 of which were determined as non-compliant samples (4.8%). Particularly high concentrations of AFM1, exceeding 200 µg/kg, were found in four samples, three raw milk and one pasteurized. Regarding this risk exposure study, only the MOE values obtained under "high exposure scenario" were lower than 10,000, while those calculated from the overall mean values resulted as not of concern. [ABSTRACT FROM AUTHOR]

Featured Application: This work offers an overview of the quality of fresh fish and fish products consumed in Puglia and Basilicata regions (south part of Italy), analyzing their histamine content. Indeed, the recent Italian reports released by the Italian Ministry of Health, concerning the official food control activities, indicated the biogenic amines (mainly histamine) as the analyte class with the highest percentage of non-compliant samples, among chemical contaminants. The histamine concentrations detected were elaborated for each type of seafood analyzed, obtaining useful information on the overall quality of fish products. The exposure assessment was also developed. This elaboration gave useful parameters also for other scientists who wish to carry out more extensive risk assessment studies. Histamine contamination was evaluated on 474 batches (3130 determinations) of fish products collected in Puglia and Basilicata (southern part of Italy) during the years 2015–2019, using a high-throughput two-tier approach involving a screening (ELISA test) and confirmatory method (HPLC/FLD with o-phthalaldehyde derivatization). Histamine concentration >2.5 mg kg−1 was detected in 51% of total batches with the 2.5% of non-compliance. Except for two samples of fresh anchovies, all non-compliant samples were frozen, defrosted and canned tuna. Among 111 fresh tuna batches, 9 had a content of histamine between 393 and 5542 mg kg−1, and scombroid poisoning cases were observed after their consumption. Good quality canned tuna and ripened anchovies sold in Italy was observed. Furthermore, the analysis of the processing technology and storage practice critical points were reported in this study, with useful considerations to minimize the histamine risk for consumers. Finally, based on these results, several considerations about risk exposure were reported. [ABSTRACT FROM AUTHOR]

A sensitive and selective analytical method for the simultaneous separation and quantitative determination of aflatoxins B1, B2, G1 and G2 in foodstuffs and materials for feed has been validated. The method is based on high performance liquid chromatography with on-line post-column photochemical derivatization and fluorescence detection. The chromatographic separation of aflatoxins was accomplished using a C18 column eluted with an isocratic mobile phase consisting of water, methanol and acetonitrile. The sample preparation required a simple extraction of aflatoxins with MeOH/H2O (80:20, v/v) and a purification step by immunoaffinity column cleanup. The total analysis time, including sample preparation and chromatographic separation, did not exceed 40 min with a run time of 10 min. The on-line photochemical derivatization ensures better results in terms of simplicity, sensitivity and reproducibility with respect to chemical derivatization techniques, and provides an increase of the peak resolution and an extent of automation in comparison with the electrochemical ones. The procedure for the determination of aflatoxins in food samples and cereals for animal consumption was extensively validated following Regulation (EC) No. 882/2004. Detection limits in wheat bran samples of 0.08 µg kg-1 for AFB1, 0.02 µg kg-1 for AFB2, 0.16 µg kg-1 for AFG1 and 0.04 µg kg-1 for AFG2 were attained. The method allows high recovery with mean values ranging from 72 to 94% and it satisfies the necessary requirements for sensitivity, linearity, selectivity, precision and ruggedness, demonstrating the conformity of the method with provisions of Regulation (EC) No. 401/2006. [ABSTRACT FROM AUTHOR]

A high-performance liquid chromatographic method with on-line post-column photochemical derivatization and fluorimetric detection for the simultaneous separation and quantitative determination of aflatoxin (AF) B(1), B(2), G(1), and G(2) in foodstuffs and feed materials is reported.The chromatographic separation is accomplished by using a C(18) column eluted with an isocratic mobile phase consisting of water, methanol, and acetonitrile. The sample preparation requires a simple extraction of aflatoxins with a mixture of water and methanol, and a purification step by immunoaffinity column clean-up. The total analysis time, including sample preparation and chromatographic separation, does not exceed 40 min with a run time of 10 min. The procedure for the determination of aflatoxins in food samples and cereals for animal consumption has been extensively validated, in agreement with Regulation (EC) No. 882/2004, demonstrating the conformity of the method with provisions of Regulation (EC) No. 401/2006 in terms of sensitivity, linearity, selectivity, and precision.

A sensitive and selective analytical method for the quantitative determination of fumonisins B(1) (FB(1)) and B(2) (FB(2)) in maize-based foods for direct human consumption is described. The method, based on high-performance liquid chromatography and fluorescence detection, presents a rapid and automated online post-column derivatization, performed with o-phthalaldehyde and N,N-dimethyl-2-mercaptoethylamine (Thiofluor™). A complete separation of fumonisins is achieved in less than 13 min by using a C18 column and a gradient elution. Fumonisins are extracted from the sample with a mixture of water, acetonitrile, and methanol. The filtered extract is purified by immunoaffinity column and FB(1) and FB(2) are eluted with methanol. The method has been successfully validated, and performances comply with -criteria of the Regulation EC No 401/2006.

A sensitive and reliable analytical method based on immunoaffinity chromatography cleanup followed by HPLC separation and fluorimetric detection is described for the quantitative determination of aflatoxin M(1) in milk. The chromatographic separation is accomplished by using a C18 column and a gradient elution with methanol, acetonitrile, and water. No extraction solvent process is required and a minimal milk sample cleanup is performed by a direct loading of the immunoaffinity columns and elution with methanol. The method has been successfully validated according to Decision EC No 657/2002 by using the conventional validation approach. The results of the validation process demonstrate the agreement of the method with the provisions of Regulation EC No 401/2006.