Digital Re-print - July | August 2014

Mycotoxins - How to analyse and reduce the hazard to humans and animals

www.gfmt.co.uk

Grain & Feed Milling Technology is published six times a year by Perendale Publishers Ltd of the United Kingdom.All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies, the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis of information published. ©Copyright 2014 Perendale Publishers Ltd. All rights reserved. No part of this publication may be reproduced in any form or by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1466-3872

Innovations for a better world.

Success comes with the original product.

Quality always pays off. Bühler is setting standards in the grain processing industry for more than 150 years. Whether you grind wheat, corn, rye, oat, buckwheat, soy, or malt grain – our processes and equipment are finely tuned to get the most from your grain. And this kind of process quality quickly pays off. The highest flour yields and best product quality ensure fast return on investment. www.buhlergroup.com/milling

MDDR_2014_en_de_es.indd 1 07.05.2014 13:36:20

Mycotoxins are a major hazard to humans and animals, often being found in a wide range of food and

feed samples and causing cancer as a result of ingestion of contaminated commodities.

To reduce the risk, there is the need to control mycotoxins in certain types of susceptible cereals and foods. Mycotoxin analysis through the use of immunoaffin-ity column clean-up prior to HPLC is the worldwide method of choice.

These methods have been validated by inter-laboratory studies, in many cases using immunoaffinity columns supplied by R-Biopharm Rhône Ltd. They have been shown to improve analysis, enable laborato-ries to meet legislation and to ensure a safe and wholesome food supply.

Mycotoxins in generalAs human life expectancy contuinues to

rise, this results in more disease and the dis-covery of different types of cancer. In many cases, naturally occuring constituents have been found to be the source of the disease. Mycotoxins have not only been shown to cause cancer, but also nephritis, hepatic dis-eases, various hemorrhagic syndromes, and immune and neurological disorders.

Mycotoxins are toxic secondary metabo-lites produced by moulds on food and feed products. Currently, at least 200 species have been identified and have been shown to produce more than 100 different myco-toxins.

The most common and toxic mycotox-ins are aflatoxins, ochratoxin, zearalenone, deoxynivalenol, fumonisins, citrinin, patulin, T-2 and HT-2, which are produced by Aspergillus, Fusarium and Penicillium strains. Different mycotoxins can be produced by the same fungus. Hence different mycotox-ins can appear in one commodity.

Aflatoxins are produced by the Aspergillus strain, which is found in tropi-cal regions and affects various nuts, maize, dried fruits and spices. Ochratoxin is found in cold temperatures but affects similiar commodities to aflatoxins and can be pro-duced by Aspergillus and Penicillium strains. Zearalenone, deoxynivalenol, fumonisins, citrinin, T-2 and HT-2 are Fusarium toxins and mostly appear in cereals.

Whereas patulin is limited to be found on soft fruits, fruit juices and purées.

Moulds can infect agricultural crops during crop growth, harvest, storage or processing. The growth of the mould is not necessarily related to the formation of mycotoxins and the stability of the toxin means that they may be present in food when the fungi are no longer present. Mycotoxins are produced depending on the conditions such as humidity, moisture, temperature and rainfall. Another route can be during harvest or transportation by using the same equipment or trucks to handle or transport different food and feed commodi-ties, such as crops, fruits and spices.

Only one of those commodities needs to be infected and the spread of mycotoxins

is in progress. Mycotoxins can easily pass along the foodchain when animals are fed with contaminated feed. Studies show that cows which are infected with aflatoxin (B1, B2, G1 and G2) through their feedstuff produce milk which is contaminated with aflatoxin M1.

The presence of mycotoxins in the food chain is a major concern and therefore needs to be controlled. There is a growing awareness of the potential hazards of these substances, which can cause severe toxic effects at relatively low levels in food and feed. They can also cause illness and even death in humans and other mammals.

Legal basis for food controlMany countries around the world have

prescribed methods of food analysis, where the legal basis for food control is that all laboratories should use exactly the same method.

In China there are published official GB methods and these cross reference the relevant international standards, such as International Standards Organisation (ISO) methods from which they are often based. Official methods have invariably been tested by an inter-laboratory validation study to demonstrate that different laboratories using different equipment can achieve comparable results when analysing the same samples.

These inter-laboratory studies are based on the results from a minimum of eight dif-ferent laboratories and establish the method performance in terms of the recovery,

MycotoxinsHow to analyse and reduce the hazard to humans and animals

24 | July - August 2014 GRAIN&FEED MILLING TECHNOLOGYF

Innovations for a better world.

Success comes with the original product.

Quality always pays off. Bühler is setting standards in the grain processing industry for more than 150 years. Whether you grind wheat, corn, rye, oat, buckwheat, soy, or malt grain – our processes and equipment are finely tuned to get the most from your grain. And this kind of process quality quickly pays off. The highest flour yields and best product quality ensure fast return on investment. www.buhlergroup.com/milling

MDDR_2014_en_de_es.indd 1 07.05.2014 13:36:20

precision and limits of detection and quan-tification. These parameters are known as the method performance characteristics, and indicate the performance that any laboratory can expect to achieve when closely following the method. The most widely known Official Methods are those of AOAC International, ISO, and the European Committee for Standardisation (CEN) standards.

Organisations such as AOAC International and CEN have a policy that methods are written without reference to specific consumables or other products, as these organisations do not wish to give the appearance of endorsing specific companies.

There are currently some 13 different CEN standards covering aflatoxin B1, afla-toxin M1, ochratoxin A, zearalenone, fumon-isins and patulin in a wide variety of foods (http://standards.cen.eu/). Of these official standards, eight methods employ immunoaf-finity columns for extraction and clean-up prior to analysis by HPLC. Immunoaffinity column HPLC official methods have been published covering cereals and cereal prod-ucts, nuts and nut products, dried fruit, wine, beer, baby food and infant formula.

The attraction of this approach is that irrespective of the matrix,

because of the high degree of specificity of the extraction and clean-up, comparable method performance can be achieved for almost any food or animal feed. The immu-noaffinity columns work particularly well for

pigmented matrices such as spices or other-wise complex matrices where background co-extractives need to be removed.

Immunoaffinity column Although the specific brand of immunoaf-

finity column is not specified in the CEN standard, the minimum performance of the column is given in terms of the column capacity and the recovery when myco-toxin standards are applied in solution to the immunoaffinity column. All of the immunoaf-finity columns supplied by R-Biopharm Rhône Ltd such as AFLAPREP®, OCHRAPREP®, EASI-EXTRACT® ZEARALENONE and FUMONIPREP® exceed these minimum requirements, and columns are supplied with certificates indicating their performance.

The majority of the inter-laboratory vali-dation studies from which CEN standards were generated used R-Biopharm Rhône Ltd immunoaffinity columns, which were supplied to participants who took part in the studies. The demand for further immunoaf-finity clean-up columns grows continuously.

Due to the fact that more than one fungus can infect the same commodity and certain fungal strains are able to produce more than one type of mycotoxin, modern immunoaffinity columns must be capable of detecting and quantifying more than one mycotoxin in a single run.

The product range of R-Biopharm Rhône Ltd also includes multi-toxin immunoaffin-

ity columns such as AFLAOCHRA PREP®, which allows detection of total aflatoxin as well as ochratoxin A; AOF MS-PREP®, detects aflatoxins, ochratoxin A and fumoni-sins; AO ZON PREP® detects total aflatox-ins, ochratoxin A and zearalenone; DZT MS-PREP® detects zearalenone, deoxyniva-lenol, T-2 and HT-2.

Although there are a number of different manufacturers of immunoaffinity columns for mycotoxins, it must be recognised that these products are biologically-based and each company uses different antibodies from different sources.

Thus, antibodies exhibit natural biological variability, and therefore behave differently. This means that immunoaffinity columns from different manufacturers do not perform identically nor are they of equivalent quality.

Users should therefore be aware of purchasing columns using price as a primary indicator, as cheaper poorly performing col-umns can have an adverse effect on method performance and can consequently damage a laboratory’s reputation.

R-Biopharm Rhône Ltd immunoaffin-ity columns are manufactured to ISO 9001 and the company employs an ISO 13485 quality management system. R-Biopharm Rhône Ltd products are widely used by ISO 17025 accredited laboratories in Europe and elsewhere, and are widely recognised as being reliable products of a consistently high quality.

July - August 2014 | 25GRAIN&FEED MILLING TECHNOLOGY

Analytical Solutions to check Mycotoxins in Food and Feed

R-Biopharm Rhône Ltd. • Block 10 Todd Campus, West of Scotland Science Park, Acre Road, Glasgow • Scotland G20 0XA • www.r-biopharm.com

R-Biopharm Rhône Ltd.

• RIDASCREEN® ELISAs for quantitative screening

• RIDA®QUICK Lateral Flow Tests for semi-quantitative and quantitative screening

• Immunoaffinity Columns Sample purification prior to analysis by ELISA, HPLC and LCMSMS

• Test Cards Test cards for complex matrices

• Trilogy® Reference material and standards for quality control

F

precision and limits of detection and quan-tification. These parameters are known as the method performance characteristics, and indicate the performance that any laboratory can expect to achieve when closely following the method. The most widely known Official Methods are those of AOAC International, ISO, and the European Committee for Standardisation (CEN) standards.

Organisations such as AOAC International and CEN have a policy that methods are written without reference to specific consumables or other products, as these organisations do not wish to give the appearance of endorsing specific companies.

There are currently some 13 different CEN standards covering aflatoxin B1, afla-toxin M1, ochratoxin A, zearalenone, fumon-isins and patulin in a wide variety of foods (http://standards.cen.eu/). Of these official standards, eight methods employ immunoaf-finity columns for extraction and clean-up prior to analysis by HPLC. Immunoaffinity column HPLC official methods have been published covering cereals and cereal prod-ucts, nuts and nut products, dried fruit, wine, beer, baby food and infant formula.

The attraction of this approach is that irrespective of the matrix,

because of the high degree of specificity of the extraction and clean-up, comparable method performance can be achieved for almost any food or animal feed. The immu-noaffinity columns work particularly well for

pigmented matrices such as spices or other-wise complex matrices where background co-extractives need to be removed.

Immunoaffinity column Although the specific brand of immunoaf-

finity column is not specified in the CEN standard, the minimum performance of the column is given in terms of the column capacity and the recovery when myco-toxin standards are applied in solution to the immunoaffinity column. All of the immunoaf-finity columns supplied by R-Biopharm Rhône Ltd such as AFLAPREP®, OCHRAPREP®, EASI-EXTRACT® ZEARALENONE and FUMONIPREP® exceed these minimum requirements, and columns are supplied with certificates indicating their performance.

The majority of the inter-laboratory vali-dation studies from which CEN standards were generated used R-Biopharm Rhône Ltd immunoaffinity columns, which were supplied to participants who took part in the studies. The demand for further immunoaf-finity clean-up columns grows continuously.

Due to the fact that more than one fungus can infect the same commodity and certain fungal strains are able to produce more than one type of mycotoxin, modern immunoaffinity columns must be capable of detecting and quantifying more than one mycotoxin in a single run.

The product range of R-Biopharm Rhône Ltd also includes multi-toxin immunoaffin-

ity columns such as AFLAOCHRA PREP®, which allows detection of total aflatoxin as well as ochratoxin A; AOF MS-PREP®, detects aflatoxins, ochratoxin A and fumoni-sins; AO ZON PREP® detects total aflatox-ins, ochratoxin A and zearalenone; DZT MS-PREP® detects zearalenone, deoxyniva-lenol, T-2 and HT-2.

Although there are a number of different manufacturers of immunoaffinity columns for mycotoxins, it must be recognised that these products are biologically-based and each company uses different antibodies from different sources.

Thus, antibodies exhibit natural biological variability, and therefore behave differently. This means that immunoaffinity columns from different manufacturers do not perform identically nor are they of equivalent quality.

Users should therefore be aware of purchasing columns using price as a primary indicator, as cheaper poorly performing col-umns can have an adverse effect on method performance and can consequently damage a laboratory’s reputation.

R-Biopharm Rhône Ltd immunoaffin-ity columns are manufactured to ISO 9001 and the company employs an ISO 13485 quality management system. R-Biopharm Rhône Ltd products are widely used by ISO 17025 accredited laboratories in Europe and elsewhere, and are widely recognised as being reliable products of a consistently high quality.

July - August 2014 | 25GRAIN&FEED MILLING TECHNOLOGY

Analytical Solutions to check Mycotoxins in Food and Feed

R-Biopharm Rhône Ltd. • Block 10 Todd Campus, West of Scotland Science Park, Acre Road, Glasgow • Scotland G20 0XA • www.r-biopharm.com

R-Biopharm Rhône Ltd.

• RIDASCREEN® ELISAs for quantitative screening

• RIDA®QUICK Lateral Flow Tests for semi-quantitative and quantitative screening

• Immunoaffinity Columns Sample purification prior to analysis by ELISA, HPLC and LCMSMS

• Test Cards Test cards for complex matrices

• Trilogy® Reference material and standards for quality control

F

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In this issue:

• NIR Multi Online Technology: Real-time analysis for early detection of grain quality fluctuations

• Feed Focus Pigs

• GRAPAS Technology from the GRAPAS Asia award

• Dust control with bulk bag

discharger and flexible screw conveyors

• Mycotoxins How to analyse and reduce the hazard to humans and animals

• Storage and silos special

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