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Digital Re-print - July | August 2010 Mycotoxin testing: ready for this year’s harvest?
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Managing the risks posed by mycotoxins in the food supply chain is both
complex and difficult to predict.
How will this year compare to last year for the grain and feed producers and millers?
Constant revisions are needed to our assessments. For example, the European trade organisation COCERAL have recently made reductions to their harvest forecasts for the main grain and oilseed crops in the EU.
Meanwhile the UK Met office has scrapped seasonal weather forecasts, replacing them with fortnightly and monthly ones. The task is no different for the agronomist and food technologist who need to continually assess the risks of not just single but multiple fungal toxins that need to be controlled at every stage of the supply chain.
A mycotoxin-based HACCP system
The object of this article is to present suitable options for monitoring critical con-trol points in a mycotoxin-based HACCP system. Before doing so, it is necessary to clarify certain concepts, especially regarding the characterisation of hazardous mycotoxins.
Although mycotoxins are chemical com-pounds that appear as residues in com-modities, ingredients, feeds and food, they are considered biological hazards rather than chemical hazards because their presence is a direct consequence of fungal contamination at some point in the system.1,2
Cereals and nuts are the most sensitive foods to mycotoxin contamination.
Nonetheless, these contaminants can also be detected in products of animal origin such as milk, meat, eggs, and plant derived products such as coffee, wine, dried fruit, etc. The vari-ety of commodities, feeds and foods that can be contaminated by mycotoxins is as vast as the type of contaminating mycotoxins (more than 300 are known).
Therefore, establishing a model for a mycotoxin control system in the food supply chain is no simple task.
Monitoring methods: fungal or mycotoxin analysis?
As already mentioned, despite being chemical substances mycotoxins are consid-ered biological hazards. This could lead us to believe that the correct methods for monitor-ing critical control points (CCP) are those that detect, quantify, identify and classify fungi.
However, we must consider that in many cases the different processes that eliminate fungi from commodities / ingredients will result in mycotoxins remaining throughout the food processing chain because of their chemical stability.
In other situations, potential mycotoxigenic fungi might be detected but are not produc-ing toxins due to them being a nonproducing
strain, the nutrient source being inadequate, or because the environmental conditions are not favourable for generating the mycotoxin in question.
In addition to a possible lack of correlation between the presence of potentially toxi-genic fungi and mycotoxins in a particular food product, the disadvantage of these methods is the time required.
Obtaining the end results for a single isolated strain can take between two to three weeks.
However, when monitoring a CCP in the supply chain real time measurements are required. These methods are therefore more applicable to performing a hazard analysis, than to monitoring a HACCP system once it is implemented.
Method choice and samplingThe analytical method selected depends
on a number of factors such as the type of sample, the levels of mycotoxins to be detected, the control point to be monitored and the availability of technological, eco-nomical and human resources to perform the determination.3
For all such methods the first step of sample collection and preparation is the most critical to determining what is detected and what is not.
In fact, nearly 90 percent of the error associ-ated with mycotoxin assays can be attributed to how the original sample was collected. This is because only a few percent of kernels in a contaminated lot may contain mycotoxins, and these contaminated kernels are usually not evenly distributed within the lot of grains. 4,5,6,7
Mycotoxin testing:ready for this year's harvest?
by Richard Fielder, Romer Labs, UK
Key Analytical Questions :Test on-site or outsource ?Sampling: how & how many ?Which: toxins, test, samples ?What target levels for action ?How much will it cost ?How reliable ?
Grain&feed millinG technoloGy26 | July - august 2010
FeatureMycotoxin testing
Testing for mycotoxins generally consists of three steps:
1: several small samples are taken at random from the lot and pooled into one larger ‘lot sample’
2: the entire lot sample is ground to a fine particle size and a representa-tive subsample, the ‘analytical sam-ple’, is removed for analysis and
3: the mycotoxins are extracted from the analytical sample and finally determined
From the risk perspective, if the producer wishes to avoid the chance of reject-ing an acceptable lot (Producer Risk), sampling may need optimising.
If the producer wishes to avoid the chance of
wrongful acceptance (Consumer Risk), sampling may need increasing. The choice of method can therefore be influenced by the number of sam-ples to be tested. When there is a set amount of money available, the decision is whether to choose a low-cost, on-site test capable of analys-ing many samples versus testing fewer samples by more expensive outsourced analysis.
Mycotoxin methods for HACCP monitoring
Some of the fastest and most affordable methods used are generally those based on ELISA technology.
ELISAs utilise antibody reactions in wells to provide quantitative results and are suit-able for many sample types. In common with all methods, it is important to ensure that the individual ELISA kit is validated for the mycotoxin and commodity in question, so it is ‘fit for purpose’.
Romer Labs® has over 80 validations for its AgraQuant® ELISA kits with different sample matrices. After sample preparation the ELISAs take between 15 and 25 minutes to perform and are frequently used by industries for
monitoring their CCPs, with results compara-ble to more specialist testing by HPLC.8,9
There are broadly two other types of commercially available methods for detecting mycotoxins that provide even faster determi-nation, which are very useful especially at the point of reception for raw materials.
Firstly, there are those based on detecting the natural fluorescence of the toxins, such as the FluoroQuant® Afla test kit. This test has just been adopted for the next five years by USDA’s grain inspection (FGIS) to test corn.
FluoroQuant® is simple and quick to perform, taking just five minutes after sample extraction using sample clean-up columns and a reader.
Secondly, there are other immunological-based methods (like ELISA), that are per-formed using a strip type format which indi-cates the presence or absence of a particular mycotoxin with reference to a cut-off point.
Romer Labs® has the AgraStrip® series of lateral flow devices (LFDs) for determining aflatoxins and deoxynivalenol (DON) in dif-ferent products such as corn, raisins, soy, nuts, wheat, etc.
Three cut-off levels are available for afla-
Challenges of Mycotoxins :Predicting harvest conditions Assessing the risks from single and multiple toxins Limiting their financial impact Meeting new legal safety limits Maximising international trade
Grain&feed millinG technoloGy July - august 2010 | 27
Feature Mycotoxin testing
Romer Labs UK Ltd - Ireland & UKTel: +44 845 519 5010, Fax: +44 1745 827150
e-Mail: [email protected]
®
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Design of sampling plans for mycotoxins in food
and feeds. Natural Toxins, 3, 257-262.
5. Richard J (2000). Sampling and Sample Preparation
for Mycotoxin Analysis. RomerTM Labs’ Guide to
Mycotoxins, 2.
6. Whitaker TB, Dowell FE, Hagler WM, Giesbrecht
FG, and Wu J (1994). Variability associated
with sampling, sample preparation and chemical
testing of farmers’ stock peanuts. J Assoc Off Anal
Chem Intl, 77, 107-116.
7 Whitaker TB (2003). Standardisation of mycotoxin
sampling procedures: an urgent necessity. Food
Control, 14, 233-237.
8. Zheng Z., Humphrey C., King R.S., Richard J.L. Validation
of an ELISA test kit for the detection of total aflatoxins
in grain and grain products by comparison with HPLC.
Mycopathologia (2005) 159: 255-263.
9. Zheng Z., Hanneken J., Houchins D., King R.S.,
Lee P., Richard J.L. Validation of an ELISA test
kit for the detection of Ochratoxin A in several
foods commodities by comparison with HPLC.
Mycopathologia (2005) 159: 265-272.
10. Zheng Z., Richard J.L., Binder H. A review of
rapid methods for the analysis of mycotoxin.
Mycopathologia (2006) 161: 261-273.
11. Zheng, Z.M. Validation Report of AgraStripTM
Total Aflatoxin Test, Romer Labs study number
SIN-2005- 01, 2005.
12. G. Häubl . F. Berthiller . R. Krska . R. Schuhmacher.
Suitability of a fully 13C isotope labeled internal
standard for the determination of the mycotoxin
deoxynivalenol by LC-MS/MS without clean up.
Anal Bioanal Chem (2006) 384: 692–696
laboratories with analytical systems that can certify the value of the samples.
This is also performed when want-ing to confirm results that are very close to the acceptance or rejection level for the critical limits established. These labo-ratories use chromatographic techniques, HPLC, TLC, LC-MS/MS, GC-ECD, which are highly sensitive and precise. If pos-sible, the laboratory should be accredited (for example ISO17025) or have a good Quality Assurance system to ensure that their results are accurate and that trained analysts are competent to perform the method employed.
Romer Labs® operates laboratories on three continents, in Singapore, the USA and Austria, all of which are certified (or are in process of certification) under the interna-tional norm ISO17025 and have the most advanced technology, including LC-MS/MS.
Preparation for 2010 harvestMonitoring mycotoxins throughout the
production chain is one safety measure. Other tools to prevent contamination
include the monitoring of environmental con-ditions that might favour the production of mycotoxigenic fungi for example, measuring water activity and temperatures.
The coming months are critical for the quality of this year’s harvest, which is why the UK’s Home Grown Cereal Authority have just launched their fusarium mycotoxin risk assessment to calculate the risk of contamina-tion, even though the levels of mycotoxins are generally very low in UK grain.
A good HACCP system has to be capable of coping with all factors that put the produc-tion chain at risk. Mycotoxins do not occur in isolation as discussed here, and need to be considered in the context of all the other food safety risks.
References:1. FAO /IAEA Training and Reference Centre for Food
and Pesticide Control (2001 Reprinted 2003)
Manual on the Application of the HACCP System
in Mycotoxin Prevention and Control. Rome.
2. D. Aldred, N.Magan, M. Olsen, Chp. 7: The use
of HACCP in the control of mycotoxins: the case
of cereals. In: Mycotoxin in Food – Detection and
Control. Woodhead Publishing Limited, England.
2004.
3. P.S. Knass, Micotoxinas en la Industria Alimentaria:
Mantener la Situación Bajo Control. Énfasis
Alimentación 6: 52-61 FLC Editores.
4. Coker RD, Nagler MJ, Blunden G, Sharkey AJ,
Defize PR, Derksen GB, and Whitaker TB (1995).
toxins in-line with legislative limits: 4ppb, 10ppb and 20ppb. This allows one to choose the most appropriate LFD according to the commodity and its end use.
For example, the AgraStrip® Afla 4ppb is used for analysing almonds exported to the European Community. Some LFDs are quantitative rather than qualitative when used with an optical reader. Romer has recently improved its AgraStrip® XReader when test-ing for DON, it provides instant (semi-) quantitative results with greater accuracy, for a wider range of commodities, using updated software.
As in the ELISA tests, it is necessary to verify that the analysed matrix is included among the validated commodities.10,11
Reliability of resultsAll quality management systems must
verify the correct functioning of methods used for monitoring and strive for improvements. There are two ways of ensuring this, which rather than being mutually exclusive, comple-ment each other and so both options should be used to verify that the analysis provide precise and accurate data for decision making.
The first option is using matrix reference materials (MRMs) or internal control samples.
These are product samples used to audit the analytical procedure as they contain a known level of contamination with the myco-toxin of interest and their certificates include the value of uncertainty of determining the mycotoxin in the product.
This reference or control sample is tested in the analytical system used, at a frequency established according to the number of analy-ses performed at that point.
Romer Labs® Biopure range offers a wide range of MRMs for several toxins at different levels of toxin contamination. These samples include a certificate of analysis which complies with ISO guides 31 & 35.12
The other option is to send samples from the different monitoring points to (reference)
Author’s note: Romer Labs is the leading global provider of niche diag-nostic tests for the agricultural, food and feed industry.
More inforMation:Romer Labs Diagnostic GmbHTechnopark 13430 TullnAustria
Tel: +43 2272 615331Fax: +43 2272 615331-11Email: off [email protected]: www.romerlabs.com
for More inforMation: Richard Fielder (Director)
Email: richard.f [email protected]: +44 845 5195010
EU Mycotoxin Legislation :No. 1881/2006 – foodNo. 105/2010 – food (New)No. 1126/2007 – foodNo. 2003/100/EC – feedNo. 401/2006 - foodRecommendations:2006/576/EC – feed2006/583/EC – cereals2003/598/EC – food
Grain&feed millinG technoloGy28 | July - august 2010
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COMMODITIES
possible Canadian, Black Sea or European losses not yet factored in, this would not make much of a dent in the huge 2010/11 surplus stock which the IGC estimated over 200m tonnes in late June. Overall, then, wheat
supplies should be more than adequate to meet demand – though some questions about quality remain to be answered by harvests in some key producing countries.
Less maize supply than thought
Two key factors caused a shift of emphasis in the maize market during the last month. One was an official update of US planted area which
conditions that were producing some worryingly low proteins. These seem to have improved somewhat since the harvest moved north under clearer skies. The US soft red winter harvest was also challenged by showers causing quality problems including greater incidence of vomitoxin in some states.
Among other key milling wheat producers, Australia still seems to be on course for its third large crop in a row with a developing La Nina climate phase expected to bring adequate rains. Argentina sowed a lot more wheat for its next crop and could expand output to about 12/14m tonnes from last year’s unsually poor 9.6m. If the growing season goes well, this important breadwheat supplier could have twice as much for export as last year’s measly 4.5m.
Turning briefly to demand, world wheat consumption is expected by the International Grains Council to rise from 649m to 658m tonnes, largely in the feed sector and especially in Asia. That should be easily accommodated if the crop reached the IGC’s projected 664m tonnes. Even if we knock off, say, 10m to 15m tonnes to account for
abandoned acres. With some trade estimates ranging down to as little as 7.3m ha (versus last year’s 9.9m), there is talk of a total wheat crop as low as 18.5m compared with recent hopes of 24.5m and last year’s 26.5m tonnes. As always with Canada, the proportion of top grade bread wheats in the total harvest will depend on weather in the run up to crop maturity and harvest. However, on current pointers, supply from this source does look likely to fall and this has already driven export prices of CWRS sharply higher.
Offsetting that slightly, the USDA did keep its US spring wheat sowing forecast at a surprisingly high 5.63m ha against trade ideas it would lop this figure. This crop is in the best condition seen for many years and, if the weather continues to co-operate, it should yield well in terms of both quantity and quality. However, the US hard breadwheat export benchmark (DNS) is still rising to reflect the threat to Canada’s crop, reaching a 19-week high of $296/tonne in mid-July.
US hard red winter bread wheat – the largest component of US wheat exports – had a shaky start to harvest under damp
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August 2010
• Mycotoxintesting:
ready for this year’s harvest?
In this issue:
• Technical design and equipment - Key to improving
feed quality and nutrition
• Factors affecting pelleting
and energy consumption
• Fusarium mycotoxins
– What’s all the fuss about?
• MakingFeedPellets
• AddedvaluebyFlourHeatTreatment
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