Mycotoxin analysis is usually carried out by high performance liquid chromatography after immunoaffinity column cleanup or in enzyme-linked immunosorbent assay tests. These methods normally involve determination of single compounds only. EU legislation already exists for the aflatoxins, ochratoxin A and patulin in food, and legislation will come into force for deoxynivalenol, zearalenone and the fumonisins in 2007. To enforce the various legal limits, it would be preferable to determine all mycotoxins by routine analysis in different types of matrices in one single extract. This would also be advantageous for HACCP control purposes. For this reason, a multi-method was developed with which 33 mycotoxins in various products could be analysed simultaneously. The mycotoxins were extracted with an acetonitrile/water mixture, diluted with water and then directly injected into a LC-MS/MS system. The mycotoxins were separated by reversed-phase HPLC and detected using an electrospray ionisation interface (ESI) and tandem MS, using MRM in the positive ion mode, to increase specificity for quality control. The following mycotoxins could be analysed in a single 30-min run: Aflatoxins B1, B2, G1 and G2, ochratoxin A, deoxynivalenol, zearalenone, T-2 toxin, HT-2 toxin, alpha-zearalenol, alpha-zearalanol, beta-zearalanol, sterigmatocystin, cyclopiazonic acid, penicillic acid, fumonisins B1, B2 and B3, diacetoxyscirpenol, 3- and 15-acetyl-deoxynivalenol, zearalanone, ergotamin, ergocornin, ergocristin, alpha-ergocryptin, citrinin, roquefortin C, fusarenone X, nivalenol, mycophenolic acid, alternariol and alternariol monomethyl ether. The limit of quantification for the aflatoxins and ochratoxin A was 1.0 microg kg(-1) and for deoxynivalenol 50 microg kg(-1). The quantification limits for the other mycotoxins were in the range 10-200 microg kg(-1). The matrix effect and validation data are presented for between 13 and 24 mycotoxins in peanuts, pistachios, wheat, maize, cornflakes, raisins and figs. The method has been compared with the official EU method for the determination of aflatoxins in food and relevant FAPAS rounds. The multi-mycotoxin method has been proven by the detection of more than one mycotoxin in maize, buckwheat, figs and nuts. The LC-MS/MS technique has also been applied to baby food, which is subject to lower limits for aflatoxin B1 and ochratoxin A, ergot alkaloids in naturally contaminated rye and freeze-dried silage samples.
Summary A total of twenty‐eight mycotoxins were surveyed in wine (red, white and rose), cider (white and rose) and their cork stoppers from eight countries. Toxins of different fungi genera were detected as follows: Alternaria (ATs: alternariol – AOH; alternariol methyl – AME) and Penicillium/Aspergillus (ochratoxin A – OTA; penicillic acid – PAC). Toxins and levels varied with the sample types and country of origin. Wine presented contamination of OTA, AOH and AME. OTA was detected in forty‐one wine samples with levels ranging from 0.01 to 0.86 μg L−1, below EU legislation. AOH and AME were detected in thirty‐three and eight of wines samples, respectively, at levels from 0.2 to 13.3 μg L−1, while no contamination was detected in ciders up to the method LOQs. Regarding the cork stoppers toxins detected, they were AOH, AME and PAC. Corks of red wine from different countries had levels of OAH and AME ranging from 5.0 to 101.0 and 2.5 to 5 μg g−1, respectively. It is necessary to pay more attention on the corks processing and cork type used in the bottles as, different from the ordinary ones, the ground bark and compressed type did not have toxins detected.
In April 1999 an amount of 2600 μg/kg DON was found in a sample breakfast cereals in the Netherlands. This event was the start of a lot of activities, which dealt with the prevention, control, health and consumer aspects of DON in food for human consumption. The Food Inspection Services started a monitoring program to measure DON in cereal products, flour and raw cereals. The National Institute of Public Health and the Environment, another part of the Ministry of Health in the Netherlands, was asked to carry out a risk analysis on DON. This was the basis for the Minister of Health to set an action limit for consumer products. She also informed Brussels and asked for a European limit. The Main Board on Agriculture set out to implement measures to be taken at harvesting, milling and bread baking industry. The Scientific Committee on Food of the EC expressed an opinion on DON in December 1999. Worldwide attention leads to discussion of a DON limit by JECFA in February 2001.In the period may 1999 until march 2002 a number of more than 1700 samples were analysed on DON. These originated from the cereal harvest of the years 1998 until 2001. The results showed a sharp decrease of DON content in samples of harvest 1999 when compared to 1998. This lower level was maintained in the 2000 and 2001 harvests. Apparently the measures taken to control the DON level succeeded to maintain values below the action limits. Despite these activities a smaller outbreak of DON appeared in 2001 in pasta products at a lower extent. This indicated that control should be done systematically, not sporadically, and at a European level, which is made possible since EC has set a limit in July 2000. Analytical results of the measurements are presented, together with the chronological order of the associated activities of national, EU and worldwide bodies on human health control. Special attention is paid to DON in bread, related to the level in flour.
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