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.

1. Digital Re-print -
July | August 2014
Mycotoxins - How to analyse and
reduce the hazard to humans and animals
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3. F 24 | July - August 2014 GRAIN&FEED MILLING TECHNOLOGY
Mycotoxins
How to analyse and reduce the hazard to humans and animals
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 general
As 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 control
Many 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,

4. GRAIN&FEED MILLING TECHNOLOGY July - August 2014 | 25
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.
R-Biopharm Rhône Ltd.
Analytical Solutions to check
Mycotoxins in Food and Feed
• 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
R-Biopharm Rhône Ltd. • Block 10 Todd Campus, West of Scotland Science Park, Acre Road, Glasgow • Scotland G20 0XA • www.r-biopharm.com
F

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How to analyse
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