Ochratoxin A in red pepper flakes commercialised in Turkey

The aim of this study was to investigate the presence of Ochratoxin A (OTA) in red pepper flakes commercialised in Turkey. A total of 75 samples were collected from different supermarkets and traditional bazaars in Istanbul during 2012–2013. OTA analysis was performed by high-performance liquid chromatography equipped with fluorescence detection after immunoaffinity column clean-up. The method was linear in the range 0.05–40 μg kg^?1 (r² = 0.9997). Twenty-seven out of 31 (87.1%) packed red pepper flake samples contained OTA at concentrations ranging from 0.6 to 1.0 μg kg^?1, whereas 100% of the unpacked red pepper flake samples contained OTA, in the range 1.1–31.7 μg kg^?1. Overall, only 4 unpacked samples (5.3%) contained OTA, with a mean value of 23.4 μg kg^?1, which is higher than the European Union limit. It is suggested that OTA content should be carefully considered in red pepper flake samples especially marketed in unpacked form.     

Effect of yearly conditions and management practices on ochratoxin A production in Sultana Seedless vineyards

Sun drying of seedless grapes (Vitis vinifera cv. Sultanina) is widely practised in the western Aegean Region providing Turkey with a significant share in the world trade of sultanas. Research was initiated in 1998 to determine the incidence of ochratoxin A (OTA), to identify the major factors resulting in contamination and to develop techniques to reduce or prevent contamination. This paper assesses OTA formation in five experimental vineyards located in Manisa province of Turkey between 1998 and 2003. The cultural practices recorded were tillage (type and timing), fertilisation, plant protection, irrigation, trellising, pruning, GA₃ application, harvest maturity and date, and drying practices (type of drying yard, dipping into alkaline solution, length of drying period). In the experimental vineyards, no OTA was found at veraison. OTA levels in grapes harvested at fresh maturity ranged between <LOD and 8?ng?g^?1, whereas in dried sultanas they were between <LOD and 35?ng?g^?1. The frequency of OTA-positive samples increased in dried grapes compared with fresh. The number and timing of tillage, number of damaged berries, and removal of damaged/infected berries prior to sun drying were identified as the major good practices that may contribute to the reduction of OTA in sultanas.     

Occurrence of ochratoxin A,fumonisin B2 and black aspergilli in raisins from Western Greece regions in relation to environmental and geographical factors

The presence of ochratoxin A (OTA), fumonisin B₂ (FB₂) and black aspergilli in raisins from Western Greece regions (Messinia, Corinthia, Achaia, Ilia and Zante Island) was investigated in relation to the different geographic and climatic conditions in the 2011 growing season. The biseriate species Aspergillus niger “aggregate” and A. carbonarius were mainly identified. The population of A. niger “aggregate” species occurred in all raisin samples at colony-forming units (CFU) concentrations significantly higher (mean 2.2 × 10⁵ CFU g^?1 homogenate) than those of A. carbonarius population (mean 4.9 × 10³ CFU g^?1 homogenate), which occurred in 80% of the raisin samples. OTA was found in 73% of the samples at levels ranging from 0.1 µg kg^?1 to 98.2 µg kg^?1, with the highest level occurring in a raisin sample from Ilia that also contained the highest level of A. carbonarius. The European Union legal limit for OTA was exceeded in 15% of the raisin samples. FB₂ was found in 29% of the raisin samples at levels ranging from 7.1 µg kg^?1 to 25.5 µg kg^?1, with 20% of the samples co-occurring with OTA. Principal-component analysis was applied to levels of mycotoxins, fungal contamination, geographical data and environmental conditions recorded in the harvesting (August) or drying (September) period. Principal-component analysis clearly indicated a good direct correlation of rainfall and relative humidity with OTA and A. carbonarius contamination. A lack of clustering was observed when A. niger and FB₂ contamination were considered. This is the first report on the co-occurrence of the mycotoxins OTA and FB₂ in dried vine fruits from Greece.     

Aflatoxins and ochratoxin A in export quality raisins collected from different areas of Pakistan

During 2012–2014, 170 samples of export quality raisins were collected from different vendors in Pakistan. The collected samples were analysed for the presence of aflatoxins (AFs) and Ochratoxin A (OTA) contamination using high-performance liquid chromatography technique. The limit of detection and limit of quantification of AFs/OTA were 0.12/0.10 and 0.36/0.30 µg kg^?1, respectively. Only 5% of the samples were contaminated with AFs, ranging 0.15–2.58 µg kg^?1 with a mean of 0.05 ± 0.26 µg kg^?1. None of the raisin samples exhibited AFs contamination above the maximum limit (ML = 4 µg kg^?1) as set by the European Union (EU). About 72% of the samples were contaminated with OTA, ranging 0.14–12.75 µg kg^?1 with a mean of 2.10 ± 1.9 µg kg^?1. However, in 95.3% of the tested samples, OTA level was lower than the ML of 10 µg kg^?1 as regulated by the EU. Apparently, a strict and continuous monitoring plan, including regulatory limits, improves food safety and quality for all types of commodities.     

Effect of post-harvest treatments on the occurrence of ochratoxin A in raw cocoa beans

Cocoa beans are the principal raw material for chocolate manufacture. Moulds have an important place in the change in the quality of cocoa beans due to their role in the production of free fatty acids and mycotoxins, namely ochratoxin A (OTA). This study investigated the impact of the key post-harvest treatments, namely the fermentation and drying methods on OTA contamination of raw cocoa beans. Analytical methods for OTA detection were based on solid–liquid extraction, clean-up using an immunoaffinity column, and identification by reversed-phase HPLC with fluorescence detection. Of a total of 104 randomly selected cocoa samples analysed, 32% had OTA contents above 2 µg kg^–1. Cocoa sourced from pods in a bad state of health had a maximum OTA content of 39.2 µg kg^–1, while that obtained from healthy pods recorded 11.2 µg kg^–1. The production of OTA in cocoa beans increased according to the pod-opening delay and reached 39.2 µg kg^–1 after an opening delay of 7 days after harvest, while 6.1 and 11.2 µg kg^–1 were observed when pods were opened after 0 and 4 days. OTA production also seemed to depend considerably to the cocoa fermentation materials. When using plastic boxes for bean fermentation, the OTA production was enhanced and reached an average OTA content of about 4.9 µg kg^–1, while the raw cocoa treated in banana leaves and wooden boxes recorded 1.6 and 2.2 µg kg^–1 on average respectively. In parallel, the OTA production was not really influenced by either the mixing or the duration of the fermentation or the drying materials.     

Ochratoxin A in traditional dry-cured meat products produced from sub-chronic-exposed pigs

The presence of ochratoxin A (OTA) was determined in traditional dry-cured meat products made from sub-chronically OTA-exposed pigs. The experimental group of pigs (n = 5) was treated with 300 µg OTA kg^–1 of feed during 30 days, whereas the control group (n = 5) remained untreated. After the household production of six types of dry-cured meat products based on traditional recipes, OTA residues were determined in final products produced from each treated and untreated animal using an immunoenzymatic technique (ELISA) and HPLC with fluorescence detection (HPLC-FD). The analytical methods showed acceptable analytical performance results and high correlation coefficients. Mean OTA concentrations ranged from 4.51 ± 0.11 µg kg^–1 in smoked ham to 6.87 ± 2.01 µg kg^–1 in home-made Slavonian sausage. The study demonstrated that pig exposure to OTA leads to the accumulation of OTA residues in muscle and adipose tissue used for the production, and consequently results in contamination of the final meat products.     

Co-occurrence of aflatoxins,ochratoxin A and citrinin in “egusi” melon (Colocynthis citrullus L.) seeds consumed in Ireland and the United Kingdom

The natural co-occurrence of aflatoxins (AFs), ochratoxin A (OTA) and citrinin (CIT) in melon seed samples obtained from retailers and households in Ireland and the United Kingdom (UK) was evaluated. AFs and OTA were determined by HPLC with fluorescence detection while CIT was analysed by HPLC-MS/MS. AFB₁ was detected in all (100%) samples (mean = 9.7 μg kg^?1; range = 0.2–66.5 μg kg^?1). Mean total AFs was 12.0 μg kg^?1 (range = 0.3–82 μg kg^?1). Commercially retailed samples showed a significantly higher AFB₁ contamination (p < 0.05) than the household samples. OTA occurred in 3 (13.6%) samples, while 4 (18.2%) were contaminated with CIT at very low levels. In this study, 68% of the melon seed samples were contaminated above the 2 μg kg^?1 EU limit for AFB₁ in oilseeds. These results highlight the need for the development of strategies to reduce AF contamination in “egusi” for human consumption.     

Reduction of ochratoxin A in dry-cured meat products using gamma-irradiation

This study investigated the efficiency of gamma (γ)-irradiation in the reduction of ochratoxin A (OTA) present in dry-cured meat products prepared from intentionally contaminated raw materials from OTA-treated pigs. OTA concentrations determined in the samples (n = 24) ranged from 25.8 μg kg^–1 in bacon to 17.8 μg kg^–1 in smoked ham. After γ-irradiation at doses of 3, 7 and 10 kGy (i.e. the doses used in the food industry), a dose-depended OTA reduction was observed; however, it was not statistically significant. The mean OTA reduction achieved with 3-, 7- and 10-kGy γ-doses was approximated to 8.5%, 13.9% and 22.5%, respectively. The storage of irradiated samples (1 month, 4°C) did not significantly affect OTA levels. Based on the correlation between the OTA reduction level and basic chemical composition of dry-cured meat samples, OTA reduction may be linked to the samples’ fat content. The results indicate that γ-irradiation can reduce OTA levels in dry-cured meat products, but only to a limited extent due to the complexity of the matrix.     

Improved sensitivity of ochratoxin A analysis in coffee using high-performance liquid chromatography with hybrid triple quadrupole-linear ion trap mass spectrometry (LC-QqQLIT-MS/MS)

A novel and sensitive method utilising high-performance liquid chromatography coupled to triple quadrupole-linear ion trap mass spectrometry (LC-QqQLIT-MS/MS) was developed in order to analyse the content of ochratoxin A (OTA) in coffee samples. The introduction of the triple-stage MS scanning mode (MS³) has been shown to increase greatly sensitivity and selectivity by eliminating the high chromatographic baseline caused by interference of complex coffee matrices. The analysis included the sample preparation procedure involving extraction of OTA using a methanol–water mixture and clean-up by immunoaffinity columns and detection using the MS³ scanning mode of LC-QqQLIT-MS/MS. The proposed method offered a good linear correlation (r² > 0.998), excellent precision (RSD < 2.9%) and recovery (94%). The limit of quantification (LOQ) for coffee beans and espresso beverages was 0.010 and 0.003 µg kg^?1, respectively. The developed procedure was compared with traditional methods employing liquid chromatography coupled to fluorescent and tandem quadrupole detectors in conjunction with QuEChERS and solid-phase extraction. The proposed method was successfully applied to the determination of OTA in 15 samples of coffee beans and in 15 samples of espresso coffee beverages obtained from the Latvian market. OTA was found in 10 samples of coffee beans and in two samples of espresso in the ranges of 0.018–1.80 µg kg^?1 and 0.020–0.440 µg l^?1, respectively. No samples exceeded the maximum permitted level of OTA in the European Union (5.0 µg kg^?1).     

Occurrence of ochratoxin A in Canadian wheat shipments, 2010–12

Randomly selected domestic and export shipments (n = 1907) of Canadian durum and other wheat that occurred between 1 January 2010 and 31 December 2012 were analysed for ochratoxin A (OTA). The majority of samples did not contain OTA above the LOQ of 1 μg kg^–1. Only 37% of samples analysed contained quantifiable OTA; the median OTA of the positive results was 2.10 μg kg^–1. Canada Western Amber Durum shipments contained OTA more frequently, and at slightly higher concentrations, than Canada Western Red Spring wheat. For both wheat classes the frequency of OTA occurrence and mean concentrations appeared to increase in the lower grades, but these increases were not statistically significant. A periodic trend of a late summer increase of mean monthly OTA concentrations in shipments appears tied to the cycle of producer deliveries of wheat to primary grain elevators.     

Natural co-occurrence of ochratoxin A,ochratoxin B and aflatoxins in Sicilian red wines

The natural occurrence of ochratoxin A, ochratoxin B, aflatoxin B₁, aflatoxin B₂, aflatoxin G₁ and aflatoxin G₂ (OTA, OTB, AFB₁, AFB₂, AFG₁, AFG₂) in red wines was investigated by HPLC/FLD after immunoaffinity column clean-up in 57 market samples produced in Sicily (Italy). The results showed a very low incidence of these mycotoxins in analysed samples, confirming the high degree of quality and safety of Sicilian red wines. The results indicated 71.9% and 64.9% positive samples for OTA and OTB respectively, with an average level of 0.13 μg l^–1, well below the European maximum permitted levels (MLs). The aflatoxin most frequently detected in the samples was AFG₁, present in 57.9% of samples, while the other aflatoxins were rarely present. Recovery experiments were carried out on eight mycotoxin-free red wines spiked with OTA, OTB, AFB₁, AFB₂, AFG₁ and AFG₂ at two different levels. The limits of detection (LODs) in wines were 0.02 µg l^–1 for OTA, 0.04 µg l^–1 for OTB, 0.03 µg l^–1 for AFG₁, AFG₂ and AFB₂, and 0.05 µg l^–1 for AFB₁. A good correlation was found, with good performances in term of precision for the method.     

Post-harvest practices linked with ochratoxin A contamination of coffee in three provinces of Cordillera Administrative Region,Philippines

One of the emerging concerns in the Cordillera Administrative Region, Philippines is ochratoxin A (OTA) contamination in coffee. During 2015 to 2016, a total of 51 Arabica (Coffea arabica) coffee samples from Benguet province and 71 Robusta (Coffea canephora var. Robusta) coffee samples from the provinces of Ifugao and Kalinga were analysed for OTA contamination. The OTA-producing fungal contaminants during drying and storage of Arabica and Robusta coffee were Aspergillus niger and Aspergillus ochraceus. Ochratoxin A was more commonly detected in Robusta coffee (36.6%) than in Arabica coffee (21.6%). Among the contaminated samples, Robusta coffee cherries in the drying yard had the highest mean OTA level (120.2 μg kg^?1, n = 10) while roasted Robusta coffee beans had the lowest mean level (4.8 μg kg^?1, n = 9). The onset of contamination of Arabica coffee occurred during storage, with a mean OTA level of 46.7 μg kg^?1 (n = 9). Roasted coffee had lower OTA content although five samples had levels >5.0 μg kg^?1. Pearson Chi-square analysis (χ²) and Fisher’s exact test revealed that several post-harvest practices involving non-removal of the husk or hull and mixing of defective coffee were significantly associated with the occurrence of OTA during drying and storage (p < 0.05). No significant associations, however, were identified during roasting. This study suggests that the post-harvest practices in Cordillera Administrative Region should focus on the removal of defective coffee in all stages of post-harvest and rapid reduction of moisture content particularly during drying.     

Chemical Composition, Antioxidant Capacity, and Sensory Quality of Pomegranate (Punica granatum L.) Arils and Rind as Affected by Drying Method

The objective of this study was to evaluate the application of: (1) freeze drying, (2) convective drying (50, 60, or 70 °C), (3) vacuum–microwave drying (240, 360, or 480 W), and (4) a combined method of convective pre-drying and vacuum–microwave finish drying in the processing of pomegranate arils and rind. The quality parameters under study included sugars and organic acids, punicalagins and ellagic acid, total polyphenols, total antioxidant activity, and sensory quality. In general, drying led to a reduction in all studied parameters; however, the behavior of arils and rind was different. Vacuum–microwave drying at 240 or 360 W was the best drying treatment for arils, while rind required freeze drying or soft conditions of convective drying (50 °C). Further research is needed to obtain proper results with convective pre-drying and vacuum–microwave finish drying of arils and rind. With proper selection of the drying protocol, high-quality dried arils will be available for consumers; these arils will be characterized by high contents of fructose (25 g 100 g^?1), phytic acid (2.2 g 100 g^?1), punicalagins (0.57 mg g^?1), total polyphenols (1.6 mg eq gallic acid g^?1), high antioxidant capacity (0.6 mg eq Trolox g^?1), and high intensities of garnet color, sweetness, sourness, and fresh pomegranate aroma. Besides, dried rind with very high contents of active compounds (123 mg g^?1 of punicalagins and 108 mg eq gallic acid g^?1) and high antioxidant capacity (26 mg eq Trolox g^?1) will be also available as functional material.     

Occurrence of ochratoxin A and toxigenic potential of fungal isolates from Spanish grapes

This paper reports the results from an extensive survey of filamentous fungi isolated from wine‐producing grapes and their capacity to produce ochratoxin A (OTA) on Czapek Yeast Autolysate agar (CYA), in order to assess their potential for producing this toxin in grapes. Grapes were sampled from four Spanish wine‐producing regions during 2001. The fungal infection in berries increased with time, reaching 100% at harvest. A total of 386 isolates of Aspergillus section Nigri and 10 of Aspergillus section Circumdati were isolated and tested for their ability to produce OTA in CYA. 21 strains produced OTA (18 Aspergillus section Nigri and 3 Aspergillus section Circumdati). Aspergillus section Circumdati isolates produced higher amounts of OTA than Aspergillus section Nigri ones, with means of 10.76 µg g^?1 CYA and 1.42 µg g^?1 CYA, respectively. Despite this, black aspergilli are believed to be highly responsible for the OTA levels found in musts and wines, as it is more widespread in grapes. Musts (n = 40) produced from the grapes collected were analysed. 15% were found to contain OTA, concentrations ranging from 0.091 to 0.813 ng ml^?1 (detection limit: 0.07 ng ml^?1), but no correlation was found with the ochratoxigenic moulds isolated from grapes. Copyright © 2004 Society of Chemical Industry     

Analysis of Ochratoxin A in Wine by High-Resolution UHPLC-MS

A method for determination of ochratoxin A (OTA) in wines using a new-solid phase extraction clean-up procedure followed with ultra performance liquid chromatography (UHPLC)-Orbitrap MS based on two scan events (full-scan Fourier transform mass spectrometer [FTMS] and higher energy-induced collision dissociation[HCD] data-dependent MS/MS) in positive ionization mode has been developed. The limit of detection (LOD) was estimated at 0.46 μg l^?1 for white wine, 0.53 and 0.54 μg l^?1 for rosé and red wines, respectively. The limit of quantification (LOQ) was estimated at 1.57 μg l^?1 in white wine, 1.77 and 1.81 μg l^?1 in rosé and red wines. Recovery experiments were carried out with spiked samples at three concentration levels (2, 5 and 10 μg l^?1). The OTA recoveries in spiked white wine samples varied from 69.6 % to 99.8 %, while the recoveries for rosé and red wine samples were in the range of 63.0–110.2 % and 63.6–103.2 %, respectively. Finally, based on the results, it is concluded that the combination of C18 cartridge with conventional particle packed columns and UHPLC LTQ-Orbitrap XL is an appropriate procedure for OTA analysis in wines.     

Evaluation of extraction methods for ochratoxin A detection in cocoa beans employing HPLC

Cocoa is an important ingredient for the chocolate industry and for many food products. However, it is prone to contamination by ochratoxin A (OTA), which is highly toxic and potentially carcinogenic to humans. In this work, four different extraction methods were tested and compared based on their recoveries. The best protocol was established which involves an organic solvent-free extraction method for the detection of OTA in cocoa beans using 1% sodium hydrogen carbonate (NaHCO₃) in water within 30 min. The extraction method is rapid (as compared with existing methods), simple, reliable and practical to perform without complex experimental set-ups. The cocoa samples were freshly extracted and cleaned-up using immunoaffinity column (IAC) for HPLC analysis using a fluorescence detector. Under the optimised condition, the limit of detection (LOD) and limit of quantification (LOQ) for OTA were 0.62 and 1.25 ng ml^–1 respectively in standard solutions. The method could successfully quantify OTA in naturally contaminated samples. Moreover, good recoveries of OTA were obtained up to 86.5% in artificially spiked cocoa samples, with a maximum relative standard deviation (RSD) of 2.7%. The proposed extraction method could determine OTA at the level 1.5 µg kg^–1, which surpassed the standards set by the European Union for cocoa (2 µg kg^–1). In addition, an efficiency comparison of IAC and molecular imprinted polymer (MIP) column was also performed and evaluated.     

Aluminium,nickel, cadmium and lead in candy products and assessment of daily intake by children in Spain

Aluminium (Al), nickel (Ni), cadmium (Cd) and lead (Pb) levels in a total of 263 samples of 12 types of candies widely consumed in Spain, were evaluated. Samples were analysed using an ICP-MS method after acidic sample mineralization. Concentration ranges of Al, Ni, Cd and Pb were 21.28–62.91 µg g^?1, 0.40–1.27 µg g^?1, 0.12–1.01 µg g^?1 and 1.03–7.14 µg g^?1, respectifgvely. Statistically significant positive correlations were calculated between concentrations of Ni–Al and Pb–Cd (p-values < 0.05). Taking into consideration the relatively high metal content, together with the high caloric density of these products, as well as high content of particular nutrients such as sugars that can induce development of certain pathologies like obesity and caries, indicate that frequent consumption of candy products is not recommended.     

Quantification of mycotoxins in vegetable oil by UPLC-MS/MS after magnetic solid-phase extraction

ABSTRACT

The detection of mycotoxin contamination in foodstuffs is highly significant for public health. Herein we report an analytical method based on magnetic solid-phase extraction (MSPE) and UPLC-MS/MS for the simultaneous determination of mycotoxins, including fumonisins B₁ (FB₁), zearalenone (ZON) and ochratoxin A (OTA), in vegetable oil. Magnetic nanoparticles coated with double layers of silicon dioxide were synthesised and found to be an effective MSPE adsorbent for mycotoxins. The proposed MSPE procedure serves not only for sample clean-up but also for mycotoxin enrichment that enhances greatly the assay’s sensitivity. Under the selected MSPE conditions, linear matrix-matched calibration curves were obtained for mycotoxins in a concentration range from 0.178 to 625 μg kg^–1. The limits of detection were 0.210 μg kg^–1 for FB₁, 0.0800 μg kg^–1 for OTA and 1.03 μg kg^–1 for ZON. The proposed MSPE UPLC-MS/MS method was applied for the determination of mycotoxins in vegetable oil samples, including maize oil, rapeseed oil and soybean oil. ZON was detected in a maize oil at 101 μg kg^–1, which is below the European Union limit of 200 μg kg^–1 in foodstuffs.     

Ochratoxin A in artisan salami produced in Veneto (Italy)

Fifty samples of artisan salami purchased in Veneto (Italy) were analysed for the determination of ochratoxin A (OTA). The analytical method, based on a sample preparation procedure with immunoaffinity columns (IACs), together with analysis by high-performance liquid chromatography with fluorescence detection (HPLC-FD), has guaranteed a high rate of recovery (about 97%), limit of detection (LOD) and limit of quantification (LOQ), respectively, of 0.06 µg kg^?1 and 0.20 µg kg^?1. OTA was detected in five samples, but only one exceeded the guideline value (1 µg kg^?1) established by the Italian Ministry of Health for pork meat and derived products. The results would seem to suggest that salami made with the traditional, non-industrial production method can be considered safe as regards contamination by OTA. However, the very high concentration observed in one sample proves that a high OTA contamination is also possible in this type of product. Thus, the controls of mycotoxin contamination must consider also salami.     

Determination of plasticisers and BPA in Sicilian and Calabrian nectar honeys by selected ion monitoring GC/MS

Twenty-six plasticisers and bisphenol A (BPA) in 39 Sicilian and Calabrian nectar honeys of different botanical south Italian origin were determined by GC-MS. Di-(2-ethylhexyl)phthalate was the most abundant plasticiser in all samples, reaching up 202.7 ± 153.1 µg kg^–1, followed by di-butylphthalate with a concentration of 40.3 ± 9.3 µg kg^–1, whereas the concentration of the other phthalates varied from not detectable to 68.2 µg kg^–1. Trace levels of adipate, sebacate and BPA were not detected in any samples. Among the different floral origin honeys, di-ethylphthalate levels were above the limit of quantification (LOQ) in orange blossom, wildflower and chestnut honey samples. Among the honeys of different geographical origin, di-methylphthalate, di-butylphthalate and di-(2-ethylhexyl)phthalate concentrations were similar. Calabrian honeys showed levels of di-ethylphthalate always lower than the LOQ, moreover di-(2-methylpropyl)phthalate levels were lower than the Sicilian ones. On the contrary, in Sicilian honeys di-(2-methylpropyl)phthalate levels were higher and di-ethylphthalate was present in 37% of samples.