Foreword: mycotoxins in a changing world

This special issue arose because of the changes in the global landscape in relation to the impact and implications of our changing climate on food security and quality, consumer habits, trade and economics, regulations and scientific thinking. The EU green paper (EC, 2007) on climate change (CC) has suggested significant hot spots in different regions where food production will be considerably affected both in quality and quantity. Indeed, a recent UNEP report on ‘Emerging Issues of Environmental Concern’ (UNEP, 2016) has included a section entitled ‘Poisoned chalice: Toxin accumulation in crops in an era of climate change’ which refers to the impact that aflatoxin contamination is having in low and middle income countries (LMICs)

Collaborative Study Report: Determination of Alternaria toxins in cereals, tomato juice and sunflower seeds by liquid chromatography tandem mass spectrometry

The Institute for Reference Materials and Measurements of the Joint Research Centre, a Directorate-General of the European Commission, organised a method validation study to evaluate the performance of a method for the simultaneous determination of five Alternaria toxins in cereals, tomato juice and sunflower seed samples. The method validation study was conducted according to the International Union for Pure and Applied Chemistry harmonised protocol. The method was used for the determination of altenuene, alternariol, alternariol monomethyl ether, tentoxin and tenuazonic acid in both naturally contaminated and fortified samples. It was based on the extraction of the test materials with an acidified methanol – water mixture, followed by solid phase extraction clean-up. The determination was carried out by reversed phase high performance liquid chromatography coupled to a triple quadrupole mass spectrometric detector. The trial involved 16 participants representing a cross section of research, private and official control laboratories from 11 EU Member States and Canada. The selection of collaborators was based on the performance in the pre-trial that was organised prior to the collaborative trial with participation of 25 laboratories. Mean recoveries reported ranged from 53% to 107%. The sample reconstitution in a water-based injection solution is thought to be responsible for the low recovery obtained for alternariol monomethyl ether, which is the least polar compound from the toxins of interest. The relative standard deviation for repeatability (RSDr) ranged from 2.0 to 34.8%. The relative standard deviation for reproducibility (RSDR) ranged from 7.7 to 49.6%, reflecting HorRat values from 0.5 to 2.4 according to the Horwitz function modified by Thompson. A correction for recovery with the data generated by spiking experiments partially improve the reproducibility performance of the method. The results highlight that the performance characteristics strongly depend on the matrix analysed, despite that fact that matrix matched calibration was used. These matrix effects can be compensated using stable isotope labelled internal standards; however, stable isotope analogues for the analysed compounds are not commercially available so far. The outcome of this study however underpins its fitness-for-purpose, which is a requirement for its formal standardisation by the European Committee for Standardization (CEN).JRC.F.5 - Food and Feed Complianc

Certified reference materials for testing of mycotoxins

This factsheet summarises the CRMs from JRC-IRMM for testing of mycotoxinsJRC.D-Institute for Reference Materials and Measurements (Geel

Report on the development of a method for the determination of Alternaria toxins and citrinin in wheat, tomato juice and sunflower seeds by liquid chromatography – tandem mass spectrometry

A new method was developed for the determination of five Alternaria toxins and citrinin in wheat, tomato juice and sunflower seed samples based on a liquid chromatography – tandem mass spectrometric determination. The optimized sample preparation involved a solid – liquid extraction with pure methanol, a derivatization step, and subsequent, solid-phase extraction purification on Strata-XL cartridges. The separation was carried out using HPLC column packed with core-shell C-18 particles. HPLC system was directly coupled to mass spectrometer via electrospray interface employed in negative mode. For the highest selectivity and sensitivity multiply reaction monitoring mode was used in the detector. The absolute recoveries (63.3% – 113.1%) and repeatability (1.2% – 13.9%) of the method, calculated from spiked samples, generated sufficient data.JRC.D.5 - Standards for Food Bioscienc

Report on the 2011 Proficiency Test of the European Union Reference Laboratory for Mycotoxins, for the Network of National Reference Laboratories: Determination of aflatoxin B1 in baby food, maize powder, animal feed and test solution

This report presents the results of a proficiency test of the EU-RL for Mycotoxins which focused on the determination of aflatoxin B1 in food and feed samples. Sixty nine participants from 28 countries registered for the exercise. Sixty-one sets of results were reported for the solution, 58 for the baby food, 67 for the maize powder and 62 for the animal feed. One laboratory did not report any results. In total about 90% of the attributed z scores were below an absolute value of two, which indicated that most of the participants performed satisfactory or better.JRC.D.5-Food Safety and Qualit

Report on the 2014 Proficiency Test of the European Union Reference Laboratory for Mycotoxins, for the Network of National Reference Laboratories - Determination of Aflatoxin B1 in Copra (Coconut powder)

This report presents the results of the PT of the EURL for Mycotoxins which focused on the determination of aflatoxin B1 (Afla B1) in coconut powder samples. Sixty-one participants from 31 countries registered for the exercise and fifty-eight sets of results were reported. Only z-scores were used for an evaluation of performance. In total 91 % of the attributed z scores were below an absolute value of two, which indicated that most of the participants performed satisfactorily.JRC.D.5-Standards for Food Bioscienc

REPORT OF THE FOLLOW-UP COLLABORATIVE STUDY Determination of the sum of Fumonisin B1 and B2 in Compound Animal Feed and Maize by Immunoaffinity Column Clean-up and High Performance Liquid Chromatography with Fluorometric Detection

The accurate determination of mycotoxins in food and feed matrices for which EU legislative limits apply require robust and reliable analytical techniques. The robustness and reliability are best shown through validation by a collaborative study. Previous collaborative studies dealing with other mycotoxins have shown that it is possible to achieve performance characteristics which are fit-for-purpose provided suitable methodology is available. As with any interlaboratory comparison homogeneity between the test units is of utmost importance. Due to the complexity of food and feed matrices particular care has to be taken during test material preparation to achieve this. Methods for the determination of Fumonisin B1 (FB1) and Fumonisin B2 (FB2) have been subject to a collaborative study in the past and the methodology used involved immunoaffinity clean-up to purify the sample extracts. Detection was afforded by derivatisation of the Fumonisins to yield fluorescent derivatives before a chromatographic separation. The reagent used was o-phtaldialdehyde and mercaptoethanol. However, pre-column derivatisation does have disadvantages related to more demanding chromatography and the instability of the derivatives. Strict time control of all processes is required to obtain adequate repeatability which necessitates the use of programmable auto liquid samplers (ALS). This may be circumvented by using post column derivatisation instead. Here the native Fumonisins are separated and reagents are added constantly to the effluent of the chromatographic column. An additional pump, a mixing Tee, and additional tubing are needed for post column derivatisation replacing the need for a sophisticated ALS. During method development it could be shown that both methods can perform equally well with respect to the requirements by EU legislation for method performance and working range. A collaborative study to validate a method for the "Determination of Fumonisin B1 and B2 in Baby Food, Breakfast Cereals and Animal Feed by Immunoaffinity Column Clean-up with High Performance Liquid Chromatography and Fluorometric Detection" failed partially because of problems with the immunoaffinity columns (IAC) used for the study. After modifications to the method protocol regarding a check of proper performance of the IAC and the sample extract clean-up we describe below the results of a repeat of the study for compound animal feed and maize.JRC.D.8-Food safety and qualit

Report on the 2007 Proficiency Test for the Determination of Ochratoxin A in Capsicum ssp (Paprika Powder)

A proficiency test was conducted with 68 laboratories from 17 EU Member States and four Third Countries. Test materials were one naturally contaminated "Ochratoxin A positive" and one "Ochratoxin A blank" capsicum material. The majority of laboratories chose to determine the ochratoxin A content by reverse-phase high-performance liquid-chromatography (RP-HPLC) with fluorescence detection against their own standard solutions as reference. Applying the modified Horwitz equation according to Thompson as a basis for the target standard deviation (22% in the case of this proficiency test), 79% of the laboratories achieved z scores of less than ¦2¦. The results were evaluated further on the basis of the returned questionnaire that each participant received. The questions asked were focussed on the fact that future method development, if necessary, could be supported by comparison of the methodologies and method procedures applied.JRC.D.8-Food safety and qualit

Report on the 2008 Proficiency Test of the Community Reference Laboratory for Mycotoxins, for the Network of National Reference Laboratories, regarding the Determination of Deoxynivalenol in a Cereal Product and a Test Solution

A proficiency test was conducted by the Community Reference Laboratory for Mycotoxins with 33 European National Reference Laboratories (NRLs) for Mycotoxins and 2 Laboratory from candidate countries, thus a total of 35 participants. Test materials were a deoxynivalenol (DON) solution in acetonitrile and three cereal test materials. Laboratories determined the DON content by either enzyme linked immuno sorbent assay (ELISA), gas chromatography (GC) or reverse-phase high-performance liquid-chromatography (RP-HPLC). One NRL did not report any results. Applying the Horwitz equation as a basis for the target standard deviation (19% in the case of this proficiency test), 27 out of the remaining 34 laboratories reported values within the z-score limit of 2 after recovery correction of the result for the DON-positive sample. Twenty-five laboratories reported results within a z-score limit of 1. Thus, 79 % of the participating laboratories performed satisfactorily in the proficiency test. No z scores were calculated for the blank material.JRC.D.8-Food safety and qualit

2014 Proficiency Test of the European Union Reference Laboratory for Mycotoxins for the Network of National Reference Laboratories - Determination of Zearalenone in Maize Oil

This report presents the results of the PT of the EURL for Mycotoxins which focused on the determination of zearalenone in maize oil. Forty-eight participants from thirty countries (among them 32 NRLs, 2 Non-EU Reference Laboratories and 13 official food control laboratories) registered for the exercise and 46 sets (Sample A and B) of results were reported. Only z-scores were used for the evaluation whether an individual laboratory underperformed. In total, 87 % of the attributed z scores were below an absolute value of two, which indicates that most of the participants performed satisfactorily.JRC.D.5-Standards for Food Bioscienc