免疫亲和柱法测定小麦中黄曲霉毒素

本文研究了用正相高效液相色谱仪附荧光检测器测定小麦中黄曲霉毒素B1、B2、C1、C2。黄曲霉毒素用甲醇－水（70＋30）提取，提取液经免疫亲和柱富集、净化，于液相色谱仪上测定。本实验采用添加法测定回收率，添加水平为0.11-3.9μg/kg时，其平均回收率为87.9%-102.5%。本方法的最低检测限为0.1μg/kg。     

A survey on the Aflatoxin M1 occurrence and seasonal variation in buffalo and cow milk from Southern Italy

This study evaluates the aflatoxin M1 (AFM1) contamination in 804 samples of raw milk from cow and buffalo, collected randomly in Campania and Calabria regions of Southern Italy over a two years period.The competitive enzyme linked immunosorbent assay (ELISA) method was used to analyze AFM1 in the samples. AFM1 levels result above the CCβ value of 0.004 μg/kg in 51 (12.3%) cow milk samples and in 28 (7.2%) buffalo milk samples. Positive results from screening analysis were confirmed by high performance liquid chromatography with fluorimetric detection (HPLC-FLD) after a procedure of centrifugation, extraction and immunoaffinity column clean-up of milk. Only one cow milk sample exceeded the maximum limit (0.05 μg/kg) set by the European Regulation.The occurrence of AFM1 contamination was significantly (p < 0.05) higher in cold season, particularly fall, than in warm season, principally spring.Our results indicate that feedstuff used in the buffalo and cow farms were not highly contaminated with aflatoxins, determining a good quality of the analyzed milk. Therefore, the AFM1 contamination of the milk does not represent a serious public health problem in both regions in Southern Italy.     

Effect of Capsicum carotenoids on growth and aflatoxins production by Aspergillus flavus isolated from paprika and chilli

The aim of this study was to determine the effect of a carotenoid mixture (Capsantal FS-30-NT), containing capsanthin and capsorubin, on growth and aflatoxins (AF) production of AF-producing Aspergillus flavus isolates.     

Analysis of aflatoxin M1 and M2 in commercial dairy products using high-performance liquid chromatography with a fluorescence detector

Aflatoxin M₁ (AFM₁) and M₂ (AFM₂) in commercial dairy products were analyzed by high-performance liquid chromatography (HPLC) with a fluorescence detector (FLD). To ensure an accurate analysis, two derivatization methods, bromination and aflatoxin–trifluoroacetic acid derivatization (ATD), were compared. The limits of detection (LODs) of the bromination method were 124.42–151.73 ng/kg, and the recovery rates were between 64 and 102%. The detection rates and concentration levels of AFM₁ were 6–74% and 14.48–270.94 ng/kg, respectively. AFM₁ was detected in 74% of milk powder samples and 36% of ice cream samples. The mean values of AFM₁ in milk powder and ice cream samples were 270.94 and 33.16 ng/kg, respectively. In the case of AFM₂, the detection rates were 2–10%, and the concentration levels were 20.62–55.67 ng/kg in milk and milk powder. Among milk and milk powder samples, ultra heat-treated (UHT) milk had lower AFM₁ contamination levels than pasteurized milk.     

Determination of aflatoxins in food using LC/MS/MS

 A liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometric method is described for the determination of aflatoxins B₁, B₂, G₁ and G₂ in food with the use of aflatoxin M₁ as an internal standard. The method works well with matrices such as those of figs and peanuts, but there are problems with spices, due to limitations of the clean-up method used. Received: 15 October 1997     

Specific detection and quantification of Aspergillus flavus and Aspergillus parasiticus in wheat flour by SYBR® Green quantitative PCR

Aflatoxins are important mycotoxins that represent a serious risk for human and animal health. These mycotoxins are mainly produced by Aspergillus flavus and Aspergillus parasiticus, two closely related species with different array of aflatoxins. In this work, two specific quantitative PCR (qPCR) assays were developed to detect and quantify both species in wheat flour using primers based on the multicopy ITS2 rDNA target sequence. The species specificity of the assays was tested in a wide range of strains of these species and others colonizing the same commodities. The sensitivity of the assay was estimated in 2.5 pg/reaction in both species. Discrimination capacity for detection and relative quantification of A. flavus and A. parasiticus DNA were analyzed using samples with DNA mixtures containing also other fungal species at different ratios. Both qPCR assays could detect spore concentrations equal or higher than 10⁶ spores/g in flour samples without prior incubation. These assays are valuable tools to improve diagnosis at an early stage and in all critical control points of food chain integrated in HACCP strategies.     

Further mycotoxin effects from climate change

Climate change will affect mycotoxins in food. The 2007 Intergovernmental Panel on Climate Change report is reinterpreted herein to account for what may occur with mycotoxins. Warmer weather, heat waves, greater precipitation and drought will have various impacts, depending on which regions of the world and mycotoxin systems are considered. The humidity issues are more complex as some areas will experience drought and others greater precipitation: in vivo data on the effects of moisture on mycotoxins in crops are more ambiguous than those for temperature. In vitro data on fungal growth and mycotoxin production may not relate directly to the situation in the field or post harvest, but are useful for base-line assumptions. The effects of climate in various regions of the world, i.e. Africa, Europe, Asia, Latin America and North America are considered in terms of mycotoxin contamination. Crops introduced to exploit altered climate may be subject to fewer mycotoxin producing fungi (the “Parasites Lost” phenomenon). Increased mycotoxins and UV radiation may cause fungi to mutate on crops and produce different mycotoxins. Whereas there is relevant information on aflatoxins, deoxynivalenol, and ochratoxin A, more mycotoxins require to be considered: Data on patulin are missing. The current paper considers uniquely ergot alkaloids. Amelioration strategies are provided. There is considerable urgency in the need to address these issues.     

Preparation and application of immunoaffinity column coupled with dcELISA detection for aflatoxins in eight grain foods

This paper described a reliable and simple analytical method for the determination of aflatoxins (AFB₁, AFB₂, AFG₁, AFG₂) in cereals, peanuts, vegetable oils and fermented foods like beer, soybean sauce and soybean paste based on immunoaffinity column (IAC) cleanup coupled with direct competitive enzyme-linked immunosorbent assay (dcELISA) detection and confirmed by ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Under the optimized conditions, the home-made IAC capacity was 293, 60, 220 and 45 ng/mL gel for AFB₁, AFB₂, AFG₁ and AFG₂, respectively. Eight grain foods were pretreated with IAC, and then the extracts were detected by the dcELISA. The results showed that the recoveries of IAC-dcELISA ranged from 71.5% to 119.8% (relative standard deviations ≤ 16.67%) that were higher than dcELISA without IAC cleanup procedure by at least 30% on average, indicating that the matrix interference of different samples could be alleviated via the pretreatment of IAC, especially for fermented foods. The developed method was also confirmed by UHPLC-MS/MS with electrospray interface in positive mode using multiple reaction monitoring. In conclusion, this assay could be used as an effective analytical method for the determination of aflatoxins in complex grain foods.     

Determination of aflatoxins B1 and B2 by adsorptive cathodic stripping voltammetry in groundnut

In this study, adsorptive stripping voltammetry was proposed for determination of aflatoxins B1 (AFB1) and B2 (AFB2) using hanging mercury drop electrode (HMDE) as the working electrode. Both aflatoxins were found to adsorb and undergo irreversible reduction reaction at the working mercury electrode. The experimental conditions were optimised by one-at-a time and experimental design to obtain the best characterised peak in terms of peak height with analytical validation of the method for each aflatoxin. The calibration curves for aflatoxins AFB1 and AFB2 were linear in the ranges of 0.4–40 ng ml⁻¹ and 0.2–70 ng ml⁻¹ with the limit of detections (LOD) 0.15 and 0.10 ng ml⁻¹, respectively. The proposed method was applied for the analysis of aflatoxins in groundnut samples and the results were compared with those obtained by the HPLC technique.