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.     

Water availability and calcium propionate affect fungal population and aflatoxins production in broiler finisher feed during storage

The aim of this study was to investigate the effects of calcium propionate, water activity (a_w) and incubation time on the total fungal count and aflatoxins B₁ (AFB₁), B₂ (AFB₂), G₁ (AFG₁) and G₂ (AFG₂) production in the broiler finisher feed. The feed was added with calcium propionate (5 g kg^–1), adjusted to 0.85, 0.90 and 0.95 a_w and stored for 28 days at 25°C, analysing for mould growth and aflatoxins production every 7 days. Analysis of variance indicated that all the factors (preservative, a_w and storage time) alone and in combination significantly (p < 0.001) affected the total fungal count and aflatoxins production in the feed. Minimum total fungal counts (1.99 × 10² CFU g^–1) were observed in calcium propionate feed at 0.85 a_w on day 1 and the highest (4.36 × 10⁹ CFUs g^–1) in control sample at 0.95 a_w on day 28 of storage. During the storage period, AFB₁ content in control samples increased from 11.35 to 73.44, from 11.58 to 81.81 and from 11.54 to 102.68 ng g^–1, whereas in preserved feed the content of B₁ increased from 11.47 to 37.83, from 11.54 to 49.07 and from 11.20 to 53.14 ng g^–1 at 0.85, 0.90 and 0.95 a_w, respectively. Similar patterns were noted for AFB₂, AFG₁ and AFG₂ contents. All the aflatoxins readily increased over storage time; however, the increase was much slower in preserved feed that contained a lower amount of available water. This study reveals that calcium propionate addition to poultry litter along with water activity amelioration is an effective tool for controlling mould incidence and aflatoxin production in poultry feed.     

Aflatoxin accumulation in whole crop maize silage as a result of aerobic exposure

BACKGROUND: Most of the maize silage stored in horizontal silos is exposed to air and can be spoiled by fungi. Potentially toxigenic fungi have been found in maize silage, and about 300 mycotoxins have been detected. Among these mycotoxins, the most harmful for feed and food safety are aflatoxins. The aim of the study was to set up a specific method to detect aflatoxins in maize silage, and to investigate whether aflatoxin contamination in maize silage depends on the level of field contamination of the crop, and whether the occurrence of aerobic spoilage during ensiling has any effect on the final contamination of the silage. RESULTS: A method for the determination of aflatoxin B₁, B₂, G₁ and G₂ in maize silage using high‐performance liquid chromagraphy with fluorescence detection has been developed and validated. Recoveries of aflatoxin B₁, B₂, G₁, and G₂ spiked over the 0.25 to 5 µg kg^?1 range averaged 74–94%. The results of laboratory scale and farm scale ensiling experiments indicated that aflatoxins could increase when silage is exposed to air during conservation or during the feed‐out phase. CONCLUSIONS: The method here proposed to detect aflatoxins in silages has proved to be sensitive and is able to detect levels of 0.1 and 0.5 ng mL^?1 for AFB₁ and AFG₁, and between 0.025 and 0.125 ng mL^?1 for AFB₂ and AFG₂. This study also provides evidence of aflatoxin accumulation in whole crop maize silage as a result of aerobic exposure. Copyright © 2011 Society of Chemical Industry     

Development of a class-specific monoclonal antibody-based ELISA for aflatoxins in peanut

Three class-specific monoclonal antibodies against aflatoxins were screened by a designed strategy in which aflatoxin G₂ was used as competitor in the screening ELISA system. With a high cross-reactivity (65%) to aflatoxin G₂, antibody 10C9 had the most similar sensitivity for five aflatoxins (AFB₁, AFB₂, AFG₁, AFG₂ and AFM₁), whose I₅₀ values were in a range of 2.1–3.2 ng ml⁻¹. So, antibody 10C9 was selected to develop an ELISA for determination of aflatoxin B₁, B₂, G₁, G₂ and total of them in peanut samples. And spiked recoveries were from 87.5% to 102.0%. The results indicate that the ELISA developed can accurately determine total aflatoxins in samples of peanuts after the simple and rapid extraction procedure.     

Occurrence of fumonisins and aflatoxins in cereals from markets of Hebei province of China

Fumonisins B₁, B₂ and B₃ (FB₁, FB₂ and FB₃) and aflatoxins B₁ (AFB₁), B₂ (AFB₂), G₁ (AFG₁) and G₂ (AFG₂) are both major mycotoxins of food concern, because of their wide range of concentration and possible co-occurrence. Therefore, a contamination survey in corn and wheat flour by liquid chromatography–tandem mass spectrometry was carried out. Quantification of fumonisins and aflatoxins was based on internal calibration (by the use of ¹³C₃₄-fumonisin) and external calibration, respectively. Fumonisins were detected in 95% of corn samples and in 7% of wheat flour samples, with the mean level (FB₁?+?FB₂?+?FB₃) of 441?µg?kg^?1 and 0.09?µg?kg^?1, respectively. Low levels of aflatoxins were detected in 37% of the samples with a mean level (B₁?+?B₂?+?G₁?+?G₂) of 0.12?µg?kg^?1. Fumonisins and aflatoxins were not detected in 29% of the samples analysed. Simultaneous occurrence of fumonisins and aflatoxins was observed in 12% of samples.     

Mycotoxin contamination of animal feedingstuff: detoxification by gamma-irradiation and reduction of aflatoxins and ochratoxin A concentrations

Mycotoxins are fungal secondary metabolites identified in many agricultural products screened for toxigenic moulds. They have been reported to be carcinogenic, teratogenic, tremorogenic, haemorrhagic and dermatitic to a wide range of organisms. With the increasing stringent regulations for mycotoxins imposed by importing countries such as those of the European Union, many cereals that are not safe for human consumption are used in formulations intended for animal feed. Gamma-rays are reported in the scientific literature to destroy ochratoxin A and aflatoxin in food crops and feed. The present study provides preliminary data for establishing the effect of dose of gamma-irradiation, ranging from 0 to 15 kGy, on aflatoxins and ochratoxin A reduction in commercial animal feed. The mycotoxin levels were determined by means of immunoaffinity clean-up (IAC) and HPLC with fluorescence detection (HPLC-FLD). The maximum reductions found at 15 kGy were 23.9%, 18.2%, 11.0%, 21.1% and 13.6% for ochratoxin A, aflatoxin B₁, aflatoxin B₂, aflatoxin G₁ and aflatoxin G₂, respectively. Results showed that the gamma-rays even at 15 kGy were not effective in the complete destruction of ochratoxin A and aflatoxins in the tested feed.     

Distribution of aflatoxins in shelling and milling fractions of naturally contaminated rice

The objective of this study was to determine the distribution of an economically important class of mycotoxins, the aflatoxins, in rice milling fractions. Rice plants grown under field production conditions are frequently infected with types of pathogenic fungi that produce toxic metabolites (mycotoxins). Paddy (seeds) rice from healthy plants in the field was collected and stored on a farm under humid, poorly ventilated conditions. Samples were milled into four fractions (hulls, brown rice, bran and white rice) and analysed for aflatoxins (B₁, B₂, G₁ and G₂) using a validated method. Rice fractions from healthy plants, which contained low levels of aflatoxins (less than 1?µg?kg^?1), were used to determine the efficiency of the extraction method. Seeds stored under poor conditions were found to be contaminated with aflatoxins B₁ and B₂ as were the fractions. The sums of AFB₁ and AFB₂ in stored paddy rice, hulls, brown rice, bran and white rice were 141, 39, 158, 367 and 56?µg?kg^?1, respectively. The ratio of aflatoxin B₁ and B₂ was about 10?:?1. AFG₁ and AFG₂ were less than 1?µg?kg^?1. Thus, brown rice contained 92.9% of the aflatoxins in paddy rice, whereas white rice contained only 27.9%.     

Aflatoxins in sunflower seeds and unrefined sunflower oils from Singida,Tanzania

A total of 61 samples comprising sunflower seeds (40) and unrefined sunflower oils (21) samples collected randomly from Singida, Tanzania were analysed by Reverse Phase-high performance liquid chromatography (RP-HPLC). 15% (6/40) of the seed samples were contaminated with aflatoxin B₁ ranging from limit of detection (LOD) to 218 ng g^?1 with three of them exceeding the European Commission/European Union (EC/EU) and Tanzania Bureau of Standards (TBS)/Tanzania Food and Drug Authority (TFDA) maximum limits of 2 ng g^?1 for AFB₁ in oilseeds. The levels of total aflatoxins (AFT) in seeds ranged from LOD to 243 ng g^?1. Other aflatoxins, except AFG₂, were also detected. For the unrefined sunflower oils, the levels of AFB₁ ranged from LOD to 2.56 ng mL^?1. About 80.9% (17/21) of the analysed oil samples contained AFB₁ of which 17.65% (3/17) exceeded the EC/EU and TBS/TFDA maximum limits of 2 ng mL^?1. Other aflatoxins were also detected in the oils. The measured levels indicate there is a need for food quality education among food processors.     

Moulds and mycotoxins in rice from the Swedish retail market

A survey of moulds and mycotoxins was performed on 99 rice samples taken from the Swedish retail market. The main objective was to study the mould and mycotoxin content in basmati rice and rice with a high content of fibre. Samples of jasmine rice as well as long-grain rice were also included. The samples were analysed for their content of ochratoxin A (high-performance liquid chromatography (HPLC)), aflatoxin B₁, B₂, G₁, and G₂ (HPLC, RIDA®QUICK), and mould (traditional cultivation methods in combination with morphological analysis). The majority of samples were sampled according to European Commission Regulation 401/2006. Subsamples were pooled and mixed before milling and both mould and mycotoxin analyses were performed on milled rice. The results showed that the majority of basmati rice (71%) and many jasmine rice samples (20%) contained detectable levels of aflatoxin B₁ (level of quantification = 0.1 µg aflatoxin kg^?1 rice). Two samples of jasmine rice and ten basmati rice samples contained levels over the regulated European maximum limits of 2 µg kg^?1 for aflatoxin B₁ or 4 µg kg^?1 for total aflatoxins. Aspergillus was the most common mould genus isolated, but also Penicillium, Eurotium, Wallemia, Cladosporium, Epicoccum, Alternaria, and Trichotecium were found. The presence of Aspergillus flavus in 21% of the samples indicates that incorrect management of rice during production and storage implies a risk of mould growth and subsequent production of aflatoxin. Rough estimates showed that high rice consumers may have an intake of 2–3 ng aflatoxin kg^?1 bodyweight and day^?1 from rice alone. This survey shows that aflatoxin is a common contaminant in rice imported to Europe.     

Assessment of exposure of broiler chicken in Brazil to mycotoxins through naturally contaminated feed

The objective of the present study was to assess the degree of exposure of broiler chicken to mycotoxins through naturally contaminated feed and the hygienic quality of feeds. For this purpose the total fungal count and occurrence of fumonisins and aflatoxins were evaluated in four feed types intended for broilers (n?=?158), collected from a poultry breeding farm in northern Paraná State, Brazil. In most feed samples (94 % pre-starter, 91 % starter, 99 % grower and 97 % finisher feeds) the total mould and yeast counts were below 1.0?×?10⁴ CFU/g, the maximum limit established to assure a good hygienic quality of the product. Fumonisins were detected in 94.9 % feed samples at mean levels ranging from 0.52 mg/kg (finisher) to 0.68 mg/kg (pre-starter and grower), while aflatoxins were detected in 72.1 % feed samples at mean levels ranging from 0.0022 mg/kg (pre-starter) to 0.0064 mg/kg (grower). The maximum estimated daily intake of fumonisin B₁ for broilers (0.057 mg/kg body weight/day) was below the Lowest Observed Adverse Effect Level (2 mg/kg bw/day). Concerning aflatoxins, most of the positive samples (97 %) showed aflatoxin levels below the maximum limit allowed by the European Commission 0.02 mg aflatoxin B₁/kg. Nevertheless, a rigorous monitoring program of the feed producing chain is essential in order to minimize human health hazards. To our knowledge, this is the first report on the degree of exposure of broilers to mycotoxins through naturally contaminated feed in Brazil.     

Determination of aflatoxins in edible oil from markets in Hebei Province of China by liquid chromatography–tandem mass spectrometry

Analysis of aflatoxin B₁ (AFB₁), B₂ (AFB₂), G₁ (AFG₁) and G₂ (AFG₂) in 76 edible oil samples (peanut oil, soybean oil, corn embryo oil and blended oil) was performed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). The oils were sampled from three areas (Shijiazhuang, Baoding and Tangshan) of Hebei Province of China. AFB₁ was detected in 22 samples representing 28.9%, followed by AFB₂ (7.89%) and AFG₁ (3.95%), while no AFG₂ contamination was detected in any samples. AFB₁ levels in oil samples ranged 0.14–2.72?µg?kg^?1 and AFB₂ ranged 0.15–0.36?µg?kg^?1, while lower levels of 0.01–0.02?µg?kg^?1 for AFG₁ were recorded. The paper is part of an on-going investigation of aflatoxin contamination in Chinese edible oils.     

Validation of a confirmatory analytical method for the determination of aflatoxins B1, B2, G1 and G2 in foods and feed materials by HPLC with on-line photochemical derivatization and fluorescence detection

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.     

Aflatoxins distribution in fractions derived from tofu production

ABSTRACT

Tofu or bean curd is obtained from soybean seeds being a widespread food product in Asia. The commodity used for its production can be contaminated with aflatoxins, which are secondary metabolites synthetised by species of Aspergillus flavus and A. parasiticus. Intake of contaminated food products causes toxic effects on consumers. The aim of this work was to study aflatoxin distribution in fractions obtained from pilot-scale tofu production with contaminated soybeans. The presence of the aflatoxins B₁, B₂, G₁ and G₂ (AFs) in soaking water, okara, whey and tofu was analysed. Aflatoxin analysis was performed by HPLC with fluorescence detection. The distribution of aflatoxins in all the analysed fractions was not a normal distribution. The liquid fractions (soaking water and whey) had less contamination than solid fractions (tofu and okara). The percentage AFB₁ remaining in nutritionally important fractions, okara and tofu, was between 6.2% and 67.7% (median = 18.1%) and 0.5% and 13.2% (median = 3.5%), respectively. AFB₂, AFG₁ and AFG₂ had a similar distribution. These results showed that throughout tofu production, AFs can be present in the products intended for human consumption.     

Occurrence of aflatoxins in selected spices in Poland

The aim of this study was to determine the contamination of aflatoxins B₁, B₂, G₁, G₂ in some selected spices (n = 84) in the Kuyavian-Pomeranian province of Poland. Aflatoxins were found in 63.1 % of the analysed samples. The presence of these compounds was confirmed in all the samples of pepper, nutmeg and turmeric. The maximum content for the sum of aflatoxins B₁, B₂, G₁, G₁ exceeding the acceptable level (10 μg/kg) was 16.91 μg/kg in one sample of nutmeg and 12.1 μg/kg in one sample of pepper, whereas in one sample of nutmeg the maximum content was equal to the regulatory limit. The lowest degree of contamination was found in black pepper.     

Mycotoxin contamination of sorghum and its contribution to human dietary exposure in four sub-Saharan countries

This research aimed at evaluating the safety, and the type, level and prevalence of mycotoxins in grain sorghum of four sub-Saharan African (SSA) countries (Burkina Faso, Ethiopia, Mali and Sudan). A multi-analyte LC-MS/MS method for quantification of 23 mycotoxins (nivalenol, deoxynivalenol, fusarenon X, neosolaniol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, diacetoxyscirpenol, roquefortine C, HT-2 toxin, alternariol, T-2 toxin, FB1, FB2, FB3, zearalenone, aflatoxin G₁, aflatoxin G₂, aflatoxin B₁, aflatoxin B₂, sterigmatocystin, OTA, altenuene, alternariol monomethylether) was applied to different sorghum matrices. Of the 1533 analysed samples, 33% were contaminated with at least one of the following mycotoxins: aflatoxins, fumonisins, sterigmatocystin, Alternaria toxins, OTA and zearalenone. Country of origin, colour, source and collection period of sorghum samples significantly influenced the type, level and prevalence of mycotoxins. Sterigmatocystin (15%), fumonisins (17%) and aflatoxins (13%) were the most prevalent. FB1 (274 ± 585 µg/kg) had the highest mean concentration followed by FB2 (214 ± 308 µg/kg) while diacetoxyscirpenol (8.12 ± 19.2 µg/kg) and HT-2 (11.9 ± 0.00 µg/kg) had the lowest concentrations. Neosolaniol, fusarenon-X, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, T-2 toxin, nivalenol and roquefortine C were not detected in any of the samples. Sudan had the lowest prevalence and mean concentration of all mycotoxins. Pink sorghum had the highest concentrations of fumonisins and aflatoxins. Mycotoxins from Aspergillus spp. and Alternaria spp. are the mycotoxins of concern in SSA grain sorghum with regard to prevalence, concentration and possible health risk from exposure. Based on the performed risk characterisation, daily consumption of sorghum containing aflatoxins, alternariol, alternariol monomethyl ether, sterigmatocystin and OTA could result in exceeding the established health-based guidance values for these toxins.     

The effect of village processing techniques on the content of aflatoxins in corn and peanuts in Zambia

The effect of village processing techniques on the aflatoxin content of corn and peanut products was investigated. In 30 trials, corn kernels were dehulled (bran removal), soaked for 24 h, washed and dried before grinding into flour and boiling in water to a thick consistency (Nshima). Shelled peanuts were either dry-roasted as whole kernels or ground into peanut meal and cooked. Dehulling, following by 24-h soaking (steeping) and subsequent washing significantly (P<0·05) reduced the aflatoxin B₁ content of corn flour from 900 to 150 μg kg⁻¹, and similarly that of aflatoxin G₁ from 929 to 114 μg kg⁻¹. Preparation of Nshima did not result into a substantial reduction in aflatoxin content, neither did extension of the cooking duration of 2 h afford any further decontamination. Whereas boiling peanut meal yielded a moderate reduction in the content of aflatoxins B₁ and G₁, roasting whole peanut kernels greatly reduced (P<0·001) the concentrations of the toxins from that in raw kernels (AFB₁= 8600 μg kg⁻¹ and AFG₁=6200 μg kg⁻¹) to 1300 and 1200 μg kg⁻¹, respectively. These results indicate that specific processing techniques carried out in Zambian villages are effective in reducing aflatoxin carry-over into edible fractions, while others are not. © 1998 SCI.     

Aspergillus and aflatoxin in groundnut (Arachis hypogaea L.) and groundnut cake in Eastern Ethiopia

This study was conducted to assess major Aspergillus species and aflatoxins associated with groundnut seeds and cake in Eastern Ethiopia and evaluate growers’ management practices. A total of 160 groundnut seed samples from farmers’ stores and 50 groundnut cake samples from cafe and restaurants were collected. Fungal isolation was done from groundnut seed samples. Aspergillus flavus was the dominant species followed by Aspergillus parasiticus. Aflatoxin analyses of groundnut seed samples were performed using ultra performance liquid chromatography; 22.5% and 41.3% of samples were positive, with total aflatoxin concentrations of 786 and 3135 ng g^?1 from 2013/2014 and 2014/2015 samples, respectively. The level of specific aflatoxin concentration varied between 0.1 and 2526 ng g^?1 for B₂ and B₁, respectively. Among contaminated samples of groundnut cake, 68% exhibited aflatoxin concentration below 20 ng g^?1, while as high as 158 ng g^?1 aflatoxin B₁ was recorded. The study confirms high contamination of groundnut products in East Ethiopia.     

Aflatoxins in composite spices collected from local markets of Karachi,Pakistan

This survey was carried out to evaluate the occurrence of total aflatoxins (AFs; B₁+B₂+G₁+G₂) in unpacked composite spices. A total of 75 samples of composite spices such as biryani, karhai, tikka, nihari and korma masalas were collected from local markets of Karachi, Pakistan, and analysed using HPLC technique. The results indicated that AFs were detected in 77% (n = 58) samples ranging from 0.68 to 25.74 µg kg^?1 with a mean of 4.63 ± 0.95 µg kg^?1. In 88% (n = 66) samples, AFs level was below the maximum limits (ML = 10 µg kg^?1) as imposed by EU. Furthermore, 61% (n = 46) tested samples contained AFs level between 1 and 10 µg kg^?1, 9% (n = 7) exhibited AFs contamination ranged 10?20 µg kg^?1 and only 3% (n = 2) of the investigated samples contained AFs levels higher than the ML of 20 µg kg^?1 for total aflatoxins as set by the USA. It was concluded that there is need to establish a strict and continuous national monitoring plan to improve safety and quality of spices in Pakistan.     

Increased aflatoxin contamination of dried figs in a drought year

Dried figs (4917 samples) destined for export from Turkey to the European Union were collected between September and December during the very dry crop year of 2007 and tested for aflatoxins B₁, B₂, G₁ and G₂ by immunoaffinity column clean-up and reverse-phase high-performance liquid chromatography (RP-HPLC). While 32% of the samples contained detectable levels of total aflatoxins, 9.8% of them exceeded the European Union limits. Aflatoxin levels were in the range of 0.2–259.46 µg kg^?1 and 2.04–259.46 µg kg^?1 for all samples and samples that exceeded the limits, respectively. A substantial increase in the incidence of aflatoxins was observed in 2007 compared with previous years, most likely due to the drought stress, high temperatures and low relative humidity encountered during the period from January to September of that year. In 2007, the mean temperature was 1–2°C higher, there was 300 mm less total rain, and the mean relative humidity was 10–15% lower than in 2002–06. The average concentration of individual aflatoxins present in the samples was quantified to determine whether the drought conditions promoted certain types of aflatoxins. Among the contaminated samples, aflatoxin B₁ occurred in 97% of the contaminated samples, followed by G₁ in 47%, B₂ in 24%, and G₂ in 6% of samples. Concentrations of individual aflatoxins exhibited great variability among the samples but were not significantly different from those reported in previous studies, which were conducted under conditions without drought and high temperatures.     

Single-Laboratory Validation for the Determination of Aflatoxin B1, B2, G1, and G2 in Foods Based on Immunoaffinity Column and Liquid Chromatography with Postcolumn Derivatization and Fluorescence Detection

This study presents a method validation procedure for the determination of aflatoxin B₁, B₂, G₁, and G₂ in hazelnut, hazelnut paste, walnut, peanut, pistachio, corn, and wheat. The method consisting of clean-up with immunoaffinity column, high performance liquid chromatography with postcolumn derivatization and fluorescence detection was validated in accordance with Commission Regulation 2004/882/EC. The selectivity, linearity, decision limit, detection capability, detection and quantification limits, precision, recovery, ruggedness, and measurement uncertainty of the method were determined. The limit of detection and limit of quantification values (μg/kg) were: aflatoxin B₁, 0.02, 0.07; aflatoxin B₂, 0.01, 0.02; aflatoxin G₁, 0.02, 0.07; and aflatoxin G₂, 0.01, 0.03. The relative standard deviation values for the repeatability and within-laboratory reproducibility were below 4 and 5 %, respectively. The recovery values of the spiked samples ranged from 80 to 105 %. These results complied with minimum performance criteria established by regulation 2006/401/EC. Therefore, the procedure can be implemented for the routine analysis of aflatoxins in the studied matrices.