Regulations for Mycotoxins in Food: Focus on the European ... - ARI

ARI The Bullet<strong>in</strong> of the Istanbul Technical University VOLUME 54, NUMBER 4
Communicated by Dilek Heperkan
<strong>Regulations</strong> <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong> <strong>in</strong> <strong>Food</strong>: <strong>Focus</strong> on the European Union
and Turkey
Hans P. van Egmond and Marco A. Jonker
Laboratory <strong>for</strong> <strong>Food</strong> and Residue Analysis, National Institute <strong>for</strong> Public Health and the Environment,
P.O. Box 1, 3720 BA Bilthoven, The Netherlands
<strong>Regulations</strong> on mycotox<strong>in</strong>s have been established <strong>in</strong> many countries to protect the consumer from the harmful
effects of mycotox<strong>in</strong>s. Current regulations mostly concern the aflatox<strong>in</strong>s, but regulations <strong>for</strong> other mycotox<strong>in</strong>s are
now rapidly develop<strong>in</strong>g. Various factors play a role <strong>in</strong> the decision-mak<strong>in</strong>g process of sett<strong>in</strong>g limits <strong>for</strong> mycotox<strong>in</strong>s.
These <strong>in</strong>clude scientific factors such as the availability of toxicological data and survey data, knowledge about
the distribution of mycotox<strong>in</strong>s <strong>in</strong> commodities, and analytical methodology. Economical and political factors
such as commercial <strong>in</strong>terests and sufficiency of food supply have their impact as well. International <strong>in</strong>quiries on
exist<strong>in</strong>g mycotox<strong>in</strong> legislation <strong>in</strong> foodstuffs and animal feedstuffs have been carried out several times <strong>in</strong> the last two
decades, yield<strong>in</strong>g details about tolerances, legal bases, responsible authorities, and official protocols of analysis and
sampl<strong>in</strong>g. By the end of 2003, about 100 countries (cover<strong>in</strong>g approximately 85% of the world’s <strong>in</strong>habitants) had
specific regulations or detailed guidel<strong>in</strong>es <strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong> food. A focus on the mycotox<strong>in</strong> regulatory situation <strong>in</strong>
the European Union shows that <strong>in</strong> the EU limits now exist <strong>for</strong> 40 tox<strong>in</strong>/food comb<strong>in</strong>ations, and this number will
further grow <strong>in</strong> the com<strong>in</strong>g years to 50. Turkey has also detailed regulations <strong>for</strong> various mycotox<strong>in</strong>s <strong>in</strong> food. The
number of Turkish regulations is currently much less compared to the EU, and the legal limits differ <strong>in</strong> some cases.
Various <strong>in</strong>ternational and European developments take place, which are related to mycotox<strong>in</strong> regulations or have
a potential impact on further developments of these regulations. These <strong>in</strong>clude lead<strong>in</strong>g scientific conferences as
the “World Mycotox<strong>in</strong> Forum” and the “IUPAC Symposium series on <strong>Mycotox<strong>in</strong>s</strong> and Phycotox<strong>in</strong>s”, large pan-
European projects as “BioCop” and “MON ˙ IQA” and activities driven by the European Commission (EC), such as
the creation of a “Community Reference Laboratory <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong>” and a mandate of the EC to the “European
Standardization Committee” (CEN) <strong>in</strong> the field of methods of analysis <strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong> food. It is expected that
the scientific basis and transparency of the mycotox<strong>in</strong> regulations will benefit from these developments.
Keywords: Mycotox<strong>in</strong>, regulation, limit, food, Europian Union, Turkey, risk assesment
1. Introduction
All nations have the right and the duty to
protect their citizens from the harmful effects
that undesirable substances <strong>in</strong> food may cause.
Among these compounds, mycotox<strong>in</strong>s are subject
to regulations <strong>in</strong> many countries. These natural
toxicants produced by fungi may lead to serious
adverse effects <strong>in</strong> men and animals, and specific
regulations have been established to limit their
presence <strong>in</strong> food and animal feed s<strong>in</strong>ce the late
1960s. The number of countries that have mycotox<strong>in</strong>
regulations has steadily grown and <strong>in</strong> 2004
approximately 100 countries were known that had
established specific limits <strong>for</strong> various comb<strong>in</strong>ations
of mycotox<strong>in</strong>s and commodities, often accompanied
by prescribed or recommended procedures
<strong>for</strong> sampl<strong>in</strong>g and analysis [1]. The sett<strong>in</strong>g
of mycotox<strong>in</strong> regulations was mostly a national
affair until the late 1990’s. Gradually several
economic communities (e.g. EU [European
Union], MERCOSUR [Mercado Comun del Sur],
Australia/New Zealand) harmonized their mycotox<strong>in</strong>
regulations, thereby overrul<strong>in</strong>g exist<strong>in</strong>g national
regulations. Especially <strong>in</strong> the EU, with<strong>in</strong>
a short period of time, regulations have come
<strong>in</strong>to <strong>for</strong>ce <strong>for</strong> many tox<strong>in</strong>/commodity comb<strong>in</strong>ations
and many more will follow soon, to control
mycotox<strong>in</strong>s both <strong>in</strong> food and animal feed. These
developments have an impact not only on the EU
itself, but on their trad<strong>in</strong>g partners as well, and
on those countries that have pre-accession status
to jo<strong>in</strong> the EU <strong>in</strong> the future, such as Turkey.
In this article the factors <strong>in</strong>fluenc<strong>in</strong>g the constitution
of mycotox<strong>in</strong> regulations, as well as the
role of <strong>in</strong>ternational organizations <strong>in</strong> the process
of establish<strong>in</strong>g and en<strong>for</strong>c<strong>in</strong>g regulations, will be
discussed. Some general observations are presented
about the current situation with respect
to worldwide regulations <strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong> food.
The focus will then be particularly directed on
the mycotox<strong>in</strong> regulatory situation <strong>in</strong> both the

EU and Turkey. Animal feed is beyond the scope
of this article, and there<strong>for</strong>e aspects of mycotox<strong>in</strong>
regulations <strong>in</strong> feed will only sporadically be dealt
with. F<strong>in</strong>ally some conclusions and recommendations
will be made.
2. Factors <strong>in</strong>fluenc<strong>in</strong>g the constitution of
mycotox<strong>in</strong> regulations
Several factors may <strong>in</strong>fluence the establishment
of mycotox<strong>in</strong> limits and regulations. These <strong>in</strong>clude:
• availability of toxicological data of mycotox<strong>in</strong>s
• availability of exposure data of mycotox<strong>in</strong>s
• knowledge of the distribution of mycotox<strong>in</strong>
concentrations with<strong>in</strong> a lot
• availability of analytical methods
• legislation <strong>in</strong> other countries with which
trade contacts exist
• need <strong>for</strong> sufficient food supply
The first two factors provide the necessary <strong>in</strong><strong>for</strong>mation
<strong>for</strong> hazard assessment and exposure assessment
respectively, the ma<strong>in</strong> <strong>in</strong>gredients <strong>for</strong>
risk assessment. Risk assessment is the scientific
evaluation of the probability of occurrence of
known or potential adverse health effects, result<strong>in</strong>g
from human exposure to food-borne hazards.
It is the primary scientific basis <strong>for</strong> the establishment
of regulations.
3. Risk assessment
<strong>Regulations</strong> are primarily made on the basis
of known toxic effects. For the mycotox<strong>in</strong>s
currently considered most significant (aflatox<strong>in</strong>s,
ochratox<strong>in</strong> A, patul<strong>in</strong>, fumonis<strong>in</strong>s, zearalenone
and some of the trichothecenes, <strong>in</strong>clud<strong>in</strong>g
deoxynivalenol, see Table 1), the Jo<strong>in</strong>t Expert
Committee on <strong>Food</strong> Additives (JECFA), a
scientific advisory body of the World Health Organization
(WHO) and the <strong>Food</strong> and Agriculture
Organization (FAO), has evaluated their hazard
<strong>in</strong> several sessions. In February 2001, a special
JECFA session was completely devoted to mycotox<strong>in</strong>s
[2]. The mycotox<strong>in</strong>s evaluated or reevaluated
at this 56 th JECFA meet<strong>in</strong>g <strong>in</strong>cluded
fumonis<strong>in</strong>s (B1, B2 and B3), ochratox<strong>in</strong> A, deoxynivalenol,
T-2 and HT-2 tox<strong>in</strong>s, and aflatox<strong>in</strong>
Hans P. van Egmond and Marco A. Jonker
2
M1. The report addressed several concerns about
each mycotox<strong>in</strong>: explanation of the mycotox<strong>in</strong>,
metabolism, toxicological studies, and f<strong>in</strong>al evaluation.
Along with the mycotox<strong>in</strong> evaluations,
the committee put <strong>for</strong>th general considerations
on sampl<strong>in</strong>g, analytical methods, associated <strong>in</strong>take
issues and control.
In addition to <strong>in</strong><strong>for</strong>mation about toxicity, exposure
assessment is another ma<strong>in</strong> <strong>in</strong>gredient of the
risk assessment. Reliable data on the occurrence
of mycotox<strong>in</strong>s <strong>in</strong> various commodities and data
on food <strong>in</strong>take are needed to prepare exposure
assessment. The quantitative evaluation of the
<strong>in</strong>take of mycotox<strong>in</strong>s is quite difficult. In its 56 th
meet<strong>in</strong>g, JECFA stressed the importance of the
use of validated analytical methods and the application
of analytical quality assurance (see also
the section on sampl<strong>in</strong>g and analysis procedures)
to ensure that the results of surveys provide
a reliable assessment of <strong>in</strong>take [3]. In most
of the JECFA reviews of mycotox<strong>in</strong>s, the analytical
data on the levels of contam<strong>in</strong>ation were often
<strong>in</strong>adequate from developed countries and nonexistent
<strong>for</strong> develop<strong>in</strong>g countries. Because most
mycotox<strong>in</strong> contam<strong>in</strong>ation is heterogeneously distributed,
sampl<strong>in</strong>g is another important consideration
<strong>in</strong> the development of <strong>in</strong><strong>for</strong>mation on the
levels of contam<strong>in</strong>ation.
In the EU, several activities take place that
contribute to the risk assessment process of mycotox<strong>in</strong>s.
An important role is played by the European
<strong>Food</strong> Safety Authority (EFSA). EFSA is an
<strong>in</strong>dependent body of the European Commission,
established <strong>in</strong> 2002, and charged with the development
of risk assessments on issues of concern
<strong>in</strong> the food and feed supply. EFSA publishes its
advices <strong>in</strong> the <strong>for</strong>m of scientific op<strong>in</strong>ions, which
<strong>for</strong>m a ma<strong>in</strong> basis <strong>for</strong> decision mak<strong>in</strong>g <strong>in</strong> the food
and feed sector. EFSA op<strong>in</strong>ions are prepared
by high level <strong>in</strong>dependent scientists <strong>in</strong> work<strong>in</strong>g
groups under the responsibility of Scientific Panels
[4]. Op<strong>in</strong>ions about risks of mycotox<strong>in</strong>s <strong>in</strong>
food and feed are developed <strong>in</strong> the Panel on Contam<strong>in</strong>ants
<strong>in</strong> the <strong>Food</strong> Cha<strong>in</strong>. Where appropriate
<strong>for</strong> its evaluation and advisory work, the EFSA
Panel makes use of data on occurrence generated
with<strong>in</strong> SCOOP (Scientific Cooperation on Questions
relat<strong>in</strong>g to <strong>Food</strong>) programmes. SCOOP is
funded by the European Commission, and targeted
to make the best estimates of <strong>in</strong>take of contam<strong>in</strong>ants
by EU <strong>in</strong>habitants. In the 1990s, these
activities resulted <strong>in</strong> a report on the exposure as-

<strong>Regulations</strong> <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong> <strong>in</strong> <strong>Food</strong>: <strong>Focus</strong> on the European Union and Turkey
sessment of aflatox<strong>in</strong>s [5]. SCOOP reports were
later published <strong>for</strong> several other mycotox<strong>in</strong>s <strong>in</strong>clud<strong>in</strong>g:
ochratox<strong>in</strong> A [6]; patul<strong>in</strong> [7]; and several
Fusarium tox<strong>in</strong>s: trichothecenes, fumonis<strong>in</strong>s and
zearalenone [8].
4. Sampl<strong>in</strong>g and analysis procedures
The distribution of the concentration of mycotox<strong>in</strong>s
<strong>in</strong> products is an important factor to
be considered <strong>in</strong> establish<strong>in</strong>g regulatory sampl<strong>in</strong>g
criteria. The distribution can be very heterogeneous,
as is the case with aflatox<strong>in</strong>s <strong>in</strong> peanuts.
The number of contam<strong>in</strong>ated peanut kernels <strong>in</strong>
a lot is usually very low, but the contam<strong>in</strong>ation
level with<strong>in</strong> a kernel can be very high. If
<strong>in</strong>sufficient care is taken <strong>for</strong> representative sampl<strong>in</strong>g,
the mycotox<strong>in</strong> concentration <strong>in</strong> an <strong>in</strong>spected
lot may there<strong>for</strong>e be wrongly estimated.
Also, consumption of peanuts could lead to an
accidental high s<strong>in</strong>gle dose of aflatox<strong>in</strong>s, rather
than a chronic <strong>in</strong>take at a relatively low level.
Similar situations could occur with other mycotox<strong>in</strong>/commodity
comb<strong>in</strong>ations. The risk to
both consumer and producer must be considered
when establish<strong>in</strong>g sampl<strong>in</strong>g criteria <strong>for</strong> products
<strong>in</strong> which mycotox<strong>in</strong>s are heterogeneously distributed.
The design of sampl<strong>in</strong>g procedures has
been an <strong>in</strong>ternational concern <strong>for</strong> a long time, <strong>for</strong>
<strong>in</strong>stance FAO and Codex Alimentarius have been
active <strong>in</strong> this area [9,10]. Discussions <strong>in</strong> work<strong>in</strong>g
groups of these <strong>in</strong>ternational organizations are
cont<strong>in</strong>uously carried out to f<strong>in</strong>d a harmonized <strong>in</strong>ternational
approach.
Several countries around the world gradually
established detailed science-based sampl<strong>in</strong>g
plans, which take account of the heterogeneous
distribution of mycotox<strong>in</strong>s <strong>in</strong> agricultural commodities.
Examples of official sampl<strong>in</strong>g plans
<strong>for</strong> mycotox<strong>in</strong>s are those applied by the USA
[11] and by the EU <strong>for</strong> several mycotox<strong>in</strong>s [12].
Although the approaches are different, the US
peanut <strong>in</strong>dustry, <strong>in</strong> cooperation with USDA, has
recently developed an Orig<strong>in</strong> Certification Program
(OCP) with several key EU countries that
import US peanuts <strong>in</strong>to Europe. These key markets,
<strong>in</strong> a memorandum of understand<strong>in</strong>g, have
agreed to recognize the sampl<strong>in</strong>g and test<strong>in</strong>g of
US peanuts <strong>for</strong> aflatox<strong>in</strong> be<strong>for</strong>e be<strong>in</strong>g exported
to these markets [13]. Documents show<strong>in</strong>g positive
lot identification and aflatox<strong>in</strong> test results
can be used to certify that the peanuts meet EU
3
aflatox<strong>in</strong> regulations.
Legislation calls <strong>for</strong> methods of control. Reliable
analytical methods will have to be available
to make en<strong>for</strong>cement of the regulations possible.
In addition to reliability, simplicity is desired,
as it will <strong>in</strong>fluence the amount of data that
will be generated and the practicality of the ultimate
measures taken. The reliability of mycotox<strong>in</strong>
analysis data can be improved through
the use of <strong>in</strong>terlaboratory-validated methods of
analysis (e.g. methods of AOAC International
and methods, standardized by CEN [Comité Européen
de Normalisation]: the European equivalent
of ISO). In the EU, CEN methods are becom<strong>in</strong>g
<strong>in</strong>creas<strong>in</strong>gly important <strong>in</strong> mycotox<strong>in</strong> regulatory
analysis. (See also the section: Current <strong>in</strong>ternational
events, European projects and
developments).
The use of good, validated methods of analysis
is no guarantee to obta<strong>in</strong> reliable analytical
results <strong>in</strong> mycotox<strong>in</strong> determ<strong>in</strong>ation.
Analytical Quality Assurance (AQA) is another
prerequisite <strong>for</strong> adequate food law en<strong>for</strong>cement.
AQA <strong>in</strong>cludes, where possible,
the use of (certified) reference materials (e.g.
those supplied by the European Commission’s
Jo<strong>in</strong>t Research Centre/Institute <strong>for</strong> Reference
Materials and Measurements [JRC/IRMM,
see http://www.irmm.jrc.be]), and participation
<strong>in</strong> proficiency tests (e.g. FAPAS R○ (see
www.fapas.com)). Also important, especially <strong>in</strong>
free trade areas, is how en<strong>for</strong>cement bodies handle
an issue as measurement uncerta<strong>in</strong>ty. European
legislation <strong>for</strong> mycotox<strong>in</strong>s requires that
“The analytical result must be reported as x +/-
U whereby x is the analytical result and U is
the expanded measurement uncerta<strong>in</strong>ty” [12], but
specific guidance how to estimate measurement
uncerta<strong>in</strong>ty is not provided. With<strong>in</strong> the EU and
the European Free Trade Area (EFTA) the FLEP
(<strong>Food</strong> Law En<strong>for</strong>cement Practitioners) Work<strong>in</strong>g
Party “<strong>Mycotox<strong>in</strong>s</strong>” is currently deal<strong>in</strong>g with this
issue [14]. Measurement uncerta<strong>in</strong>ty is also be<strong>in</strong>g
dealt with by the European organisations
Eurachem (www.eurachem.ul.pt) and CEN. CEN
will prepare a new document on criteria of methods
of mycotox<strong>in</strong>s <strong>in</strong> the com<strong>in</strong>g years, which
<strong>in</strong>cludes measurement uncerta<strong>in</strong>ty (see also the
section:Current <strong>in</strong>ternational events, European
projects and developments).

4.1. Trade Contacts
Preferably, regulations should be brought <strong>in</strong>to
harmony with those <strong>in</strong> <strong>for</strong>ce <strong>in</strong> other countries
with which trade contacts exist. In fact, this approach
has been applied <strong>in</strong> the areas of the EU,
MERCOSUR and Australia/New Zealand, where
now harmonized regulations <strong>for</strong> some mycotox<strong>in</strong>s
exist. Strict regulative actions may lead import<strong>in</strong>g
countries to ban or limit the import of commodities,
such as certa<strong>in</strong> food gra<strong>in</strong>s, which can
cause difficulties <strong>for</strong> export<strong>in</strong>g countries <strong>in</strong> f<strong>in</strong>d<strong>in</strong>g
or ma<strong>in</strong>ta<strong>in</strong><strong>in</strong>g markets <strong>for</strong> their products. For
example, the str<strong>in</strong>gent regulations <strong>for</strong> aflatox<strong>in</strong> Bl
<strong>in</strong> animal feedstuffs, established <strong>in</strong> the EU <strong>in</strong> the
1980s and 1990s [15], led European animal feed
manufacturers to switch from groundnut meal to
other prote<strong>in</strong> sources to <strong>in</strong>clude <strong>in</strong> feeds; this had
an impact on the export of groundnut meal of
some develop<strong>in</strong>g countries [16]. The distortion
of the market caused by regulations <strong>in</strong> import<strong>in</strong>g
countries may lead to export of the less contam<strong>in</strong>ated
foods and feeds leav<strong>in</strong>g those <strong>in</strong>ferior
foods and feeds <strong>for</strong> the local market. Some countries
apply different limits <strong>for</strong> aflatox<strong>in</strong>s <strong>in</strong> certa<strong>in</strong>
products depend<strong>in</strong>g on the dest<strong>in</strong>ation.
4.2. <strong>Food</strong> Supply
The regulatory philosophy should not jeopardize
the availability of some basic commodities
at reasonable prices. Especially <strong>in</strong> the develop<strong>in</strong>g
countries, where food supplies are already limited,
drastic legal measures may lead to lack of
food and to excessive prices. For <strong>in</strong>stance, the
dramatic food security situation <strong>in</strong> parts of Africa
has led to measures that prioritise food sufficiency
above food safety. <strong>Mycotox<strong>in</strong>s</strong> are an important
problem as evidenced by occasional outbreaks of
human mycotoxicoses, such as acute aflatoxicosis
<strong>in</strong> Kenya <strong>in</strong> 2004 and 2005 [17], the role of
aflatox<strong>in</strong>s <strong>in</strong> liver cancer and stunted growth <strong>in</strong>
children <strong>in</strong> western Africa [18] and fumonis<strong>in</strong>s <strong>in</strong>
oesophageal cancer <strong>in</strong> South Africa [19]. In the
western world, where sufficiency of food is not an
issue, (low) mycotox<strong>in</strong> limits <strong>in</strong> food are en<strong>for</strong>ceable
without too many difficulties.
4.3. Synopsis
Weigh<strong>in</strong>g the various factors at the <strong>in</strong>terface
of science, food security and regulations is not a
trivial activity and common sense is a major factor
<strong>for</strong> reach<strong>in</strong>g a decision. Public health officials
are confronted with a complex problem: mycotox<strong>in</strong>s,
and particularly the carc<strong>in</strong>ogenic mycotox<strong>in</strong>s,
Hans P. van Egmond and Marco A. Jonker
4
Africa 2003
Asia/Oceania 2003
Europe 2003
Lat<strong>in</strong> America 2003
North America 2003
0 5 10 15 20 25 30 35 40 45 50
!g/kg
Figure 1. Ranges (bars) and typical limits (∆) <strong>for</strong>
total aflatox<strong>in</strong>s <strong>in</strong> food.
should be excluded from food as much as possible.
S<strong>in</strong>ce the substances are present <strong>in</strong> foods
as natural contam<strong>in</strong>ants, however, human exposure
cannot be completely prevented, and exposure
of the population to some level of mycotox<strong>in</strong>s
has to be tolerated. Despite the dilemmas, mycotox<strong>in</strong>
regulations have been established <strong>in</strong> the
past decades <strong>in</strong> many countries, and newer regulations
are still be<strong>in</strong>g drafted. Particularly <strong>in</strong> the
European Union many developments take place
<strong>in</strong> the mycotox<strong>in</strong> regulatory area, which are scientifically
underp<strong>in</strong>ned by various European activities.
5. The mycotox<strong>in</strong> regulatory situation <strong>in</strong>
the world, the European Union and
Turkey
5.1. Worldwide mycotox<strong>in</strong> regulations
Several times <strong>in</strong> the last decades (<strong>in</strong> 1981,
1987, 1995, 2003) <strong>in</strong>ternational <strong>in</strong>quiries were
held and published about regulations <strong>for</strong> mycotox<strong>in</strong>s
<strong>in</strong> food and feed. The most recent enquiry
<strong>in</strong> 2003 was carried out by the National
Institute <strong>for</strong> Public Health and the Environment,
under contract with FAO, among the Agricultural
Services of the Dutch Embassies around the
world, with the request to gather up-to-date <strong>in</strong><strong>for</strong>mation
from the local authorities on the situation
regard<strong>in</strong>g mycotox<strong>in</strong> regulations, <strong>in</strong> as many
countries of the world as possible. Where this
procedure did not lead to the desired <strong>in</strong><strong>for</strong>mation,
personal contacts were used. By the end
of 2003, data were received from 89 countries.
Together with <strong>in</strong><strong>for</strong>mation gathered <strong>in</strong> previous
<strong>in</strong>quiries, detailed <strong>in</strong><strong>for</strong>mation became available
about the existence or absence of specific myco-

<strong>Food</strong>
EUROPE
<strong>Regulations</strong> <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong> <strong>in</strong> <strong>Food</strong>: <strong>Focus</strong> on the European Union and Turkey
19,6
AFRICA
1,5
3,6
1,8
5,5
ASIA / OCEANIA
LATIN AMERICA
NORTH AMERICA
Figure 2. Average number of mycotox<strong>in</strong> regulations
per country <strong>in</strong> the various cont<strong>in</strong>ents <strong>in</strong> 2003 <strong>for</strong> food
and food products.
tox<strong>in</strong> limits and regulations <strong>in</strong> food and feed, <strong>in</strong>
119 countries. All data received were sorted by
country and by economic community (ASEAN,
Australia/New Zealand, EU, MERCOSUR). As
well, <strong>in</strong><strong>for</strong>mation was <strong>in</strong>cluded about standards
set by Codex Alimentarius. For each separate
entry, the available data were classified <strong>in</strong>to the
categories food, dairy and feed and than tabulated
<strong>in</strong> alphabetic order of the countries. The
results of the enquiry were published <strong>in</strong> detail by
FAO <strong>in</strong> an FAO <strong>Food</strong> and Nutrition Paper <strong>in</strong> English,
French, Spanish and Ch<strong>in</strong>ese languages [1].
On a worldwide basis, at least 99 countries
had mycotox<strong>in</strong> regulations <strong>for</strong> food and/or feed
<strong>in</strong> 2003, an <strong>in</strong>crease of approx. 30% compared
to 1995. Whereas <strong>in</strong> 1995 23% of the world’s <strong>in</strong>habitants
were liv<strong>in</strong>g <strong>in</strong> a region where no known
mycotox<strong>in</strong> regulations were <strong>in</strong> <strong>for</strong>ce, this percentage
has decreased to 13% <strong>in</strong> 2003, at the benefit of
slight <strong>in</strong>creases <strong>in</strong> coverage <strong>in</strong> Lat<strong>in</strong> America and
Europe, and more significant <strong>in</strong>creases <strong>in</strong> Africa
and Asia/Oceania. In fact all countries with mycotox<strong>in</strong>
regulations <strong>in</strong> 2003 have at least regulatory
limits <strong>for</strong> aflatox<strong>in</strong> B1 or the sum of aflatox<strong>in</strong>s
B1, B2, G1 and G2 <strong>in</strong> foods and /or feeds, a
situation that was also observed <strong>in</strong> 1995. In Fig.
1 the ranges and medians of limits <strong>for</strong> total aflatox<strong>in</strong>s
<strong>in</strong> food <strong>in</strong> 2003 are depicted <strong>for</strong> the various
world regions. For several other mycotox<strong>in</strong>s specific
regulations exist as well (i.e. aflatox<strong>in</strong> M1;
the trichothecenes deoxynivalenol, diacetoxyscirpenol,
T-2 tox<strong>in</strong> and HT-2 tox<strong>in</strong>; the fumonis<strong>in</strong>s
B1, B2, and B3; agaric acid; the ergot alkaloids;
ochratox<strong>in</strong> A; patul<strong>in</strong>; phomops<strong>in</strong>s; sterigmatocyst<strong>in</strong>
and zearalenone). The number of countries
regulat<strong>in</strong>g mycotox<strong>in</strong>s has significantly <strong>in</strong>creased
5
!"#$%
EUROPE
NORTH AMERICA
1,(
!,5
!,3
AFRICA
!,6
!,$
ASIA / OCEANIA
LATIN AMERICA
Figure 3. Average number of mycotox<strong>in</strong> regulations
per country <strong>in</strong> the various cont<strong>in</strong>ents <strong>in</strong> 2003 <strong>for</strong> dairy
and dairy products.
over the years. Fig. 2 shows the average number
of mycotox<strong>in</strong> regulations <strong>in</strong> food, per country,
<strong>in</strong> the various cont<strong>in</strong>ents of the world. Fig. 2
shows that European countries have, on average,
the most regulations, as compared to the other
cont<strong>in</strong>ents. In Fig. 3 a similar presentation is
given <strong>for</strong> specific regulations <strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong>
dairy products.
Compar<strong>in</strong>g the situations <strong>in</strong> 1995 and 2003, it
appears that <strong>in</strong> 2003 more mycotox<strong>in</strong>s are regulated
<strong>in</strong> more commodities and products, whereas
tolerance limits generally rema<strong>in</strong> the same or tend
to decrease. <strong>Regulations</strong> have become more diverse
and detailed with newer requirements regard<strong>in</strong>g
official procedures <strong>for</strong> sampl<strong>in</strong>g and analytical
methodology. Although techniques prescribed
<strong>in</strong> mycotox<strong>in</strong> regulations may differ, Fig.
4 shows that they are largely based on chromatographic
procedures. Th<strong>in</strong> layer chromatography,
a veteran separation technique <strong>in</strong> mycotox<strong>in</strong>
methodology is still of major importance <strong>in</strong>
regulatory analysis.
The <strong>in</strong>quiry of 2003 showed that several regulations
have been harmonised between countries belong<strong>in</strong>g
to economic communities (Australia/New
Zealand, EU, MERCOSUR), or are <strong>in</strong> some stage
of harmonisation. Whereas harmonized tolerance
limits would be beneficial from the po<strong>in</strong>t of view
of trade, this is not necessarily the case from the
po<strong>in</strong>t of view of (equal) human health protection
around the world. Risks associated with mycotox<strong>in</strong>s
depend on both hazard and exposure. The
hazard of mycotox<strong>in</strong>s to <strong>in</strong>dividuals is probably
more or less the same all over the world (although
other factors sometimes play a role as well, e.g.
hepatitis B virus <strong>in</strong>fection <strong>in</strong> relation to the haz-

TLC
LC
m<strong>in</strong>icolumn
GC/MS
GC
Hans P. van Egmond and Marco A. Jonker
ELISA
fluorometry
frequency of techniques
Figure 4. Techniques used <strong>in</strong> mycotox<strong>in</strong> regulatory
analysis. The size of the ball is proportional with the
frequency of use of the related technique (situation
worldwide [1]).
ard of aflatox<strong>in</strong>s). Exposure is not the same, because
of differences <strong>in</strong> levels of contam<strong>in</strong>ation and
dietary habits <strong>in</strong> various parts of the world.
Shephard [20] exemplified the significance of
differences <strong>in</strong> exposure with the help of some
calculations done <strong>for</strong> fumonis<strong>in</strong>s. JECFA established
a group Provisional Maximum Tolerable
Daily Intake (PMTDI) <strong>for</strong> fumonis<strong>in</strong>s B1, B2 and
B3 of 2 µg/kg body weight per day [2]. This
PMTDI is readily exceeded by <strong>in</strong>dividuals on a
maize-based diet <strong>in</strong> which maize consumption is
of the order of 400 g/person/day. Shephard calculated
that, at a contam<strong>in</strong>ation level <strong>for</strong> fumonis<strong>in</strong>s
<strong>in</strong> maize of 2000 µg/kg (a level with<strong>in</strong> the
range of common limits) dietary exposure <strong>for</strong> a
60 kg adult would be 13 µg/kg body weight/day
or 650% of the PMTDI! In the developed world,
maize <strong>in</strong>takes are less than 10 g/person/day [20]
and contam<strong>in</strong>ation levels as high as 12 mg/kg can
be consumed be<strong>for</strong>e dietary exposure exceeds the
PMTDI set by JECFA.
Based on these data Fig. 5 has been constructed
to visualize how consumption data may
<strong>in</strong>fluence the sett<strong>in</strong>g of regulatory limits. Assum<strong>in</strong>g
a tolerable daily <strong>in</strong>take (TDI) <strong>for</strong> fumonis<strong>in</strong>s
of 120 µg per person (2µg/kg body weight, 60 kg),
Fig. 5 shows where the regulatory limit should
be set, depend<strong>in</strong>g on consumption. The area under
the TDI-curve is the “safe area”, <strong>in</strong> contrast
to the area above the TDI-curve: the undesired
area. As an example: In a country, where the
maize consumption is approx. 15 g per capita
per day, a legal limit of 8 mg/kg would suffice
to prevent that the TDI is exceeded. However,
<strong>in</strong> another country, where the maize consump-
6
limit
[mg FUM/kg maize]
20
18
16
14
12
10
8
6
4
2
0
TDI curve
100 200 300 400
daily maize consumption [g]
Figure 5. The <strong>in</strong>fluence of the differences <strong>in</strong> daily
maize consumption rates by populations worldwide
on the sett<strong>in</strong>g of limits <strong>for</strong> fumonis<strong>in</strong>s (FUM).
tion is approx. 125 g per capita per day, a legal
limit of 1 mg/kg would be required to reach the
same level of protection. In reality the spread of
maize consumption <strong>in</strong> the world is even broader.
Note that the example above is simplified. E.g.
it ignores other possible food sources, which may
contribute to fumonis<strong>in</strong>s exposure. It may help,
however, <strong>in</strong> realiz<strong>in</strong>g that decision makers should
not haphazardly harmonise mycotox<strong>in</strong> limits <strong>for</strong>
harmonisation’s sake.
5.2. Mycotox<strong>in</strong> regulations <strong>in</strong> the EU and
Turkey
Tables 1,2, and 3 give an overview of the regulations
that currently exist (and those that will
soon come <strong>in</strong>to <strong>for</strong>ce) <strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong> food <strong>in</strong>
the EU and <strong>in</strong> Turkey. The data are based on <strong>in</strong><strong>for</strong>mation
published <strong>in</strong> FAO <strong>Food</strong> and Nutrition
regulations /
country
40
30 30
20
10
0
EUROPE
[39 countries]
38
<strong>Food</strong>
EU MS
[25 countries]
16
non-EU MS
[14 countries]
14
Turkey
Figure 6. Average number of mycotox<strong>in</strong> regulations
[2006] <strong>for</strong> food and food products per country <strong>in</strong> Europe,
European Union member states, non-EU countries
and Turkey.

<strong>Regulations</strong> <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong> <strong>in</strong> <strong>Food</strong>: <strong>Focus</strong> on the European Union and Turkey
Paper 81 [1], and have been updated with <strong>in</strong><strong>for</strong>mation
about newer EU regulations [21,22,23]
and about modifications <strong>in</strong> Turkish regulations
[24].
In 1998, the first EU-harmonized regulations
<strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong> food came <strong>in</strong>to <strong>for</strong>ce (several
limits <strong>for</strong> aflatox<strong>in</strong>s, <strong>in</strong>clud<strong>in</strong>g sampl<strong>in</strong>g
protocols and criteria <strong>for</strong> methods of analysis)
and have gradually expanded to various
mycotox<strong>in</strong>s <strong>in</strong> different foodstuffs. At the
time of writ<strong>in</strong>g limits have been set <strong>for</strong> 40
tox<strong>in</strong>/commodity comb<strong>in</strong>ations, whereas limits
<strong>for</strong> a further 10 tox<strong>in</strong>/commodity comb<strong>in</strong>ations
are probably com<strong>in</strong>g <strong>in</strong>to <strong>for</strong>ce soon. In addition,
new limits will probably be established <strong>in</strong>
the years to come <strong>for</strong> ergot alkaloids, and some
may be revised <strong>for</strong> aflatox<strong>in</strong>s, depend<strong>in</strong>g on EFSA
scientific op<strong>in</strong>ions, that are planned to be drafted.
In Turkey legal limits exist <strong>for</strong> the aflatox<strong>in</strong>s,
ochratox<strong>in</strong> A and patul<strong>in</strong>. Some limits are the
same as those applied <strong>in</strong> the EU. Clear differences
also exist <strong>in</strong> the regulations. This means
that several of the Turkish mycotox<strong>in</strong> regulations
would need to be modified and extended, should
the country become a member of the European
Union. In Fig. 6, a comparison is made between
the average number of mycotox<strong>in</strong> regulations <strong>for</strong>
food per country, <strong>for</strong> the whole of Europe, <strong>for</strong>
the European Union member states, <strong>for</strong> non-EU
countries and <strong>for</strong> Turkey. The data used were
based on the regulations <strong>in</strong> the EU and Turkey
as they existed per 1 July 2006 (Table 2, 3, respectively).
For the European non-EU countries
the data were taken from FAO <strong>Food</strong> and Nutrition
Paper 81 [1]. For this presentation it was
assumed that the 10 countries that jo<strong>in</strong>ed the EU
<strong>in</strong> 2004 have completely replaced their mycotox<strong>in</strong>
regulations by the newest (July 2006) EU mycotox<strong>in</strong>
regulations, and that these new member
states do not have country-specific mycotox<strong>in</strong> regulations
(anymore). The EU takes a lead when
it comes to the variety of mycotox<strong>in</strong> regulations
that exist <strong>for</strong> food. In the near future the number
of regulations will even further <strong>in</strong>crease. Similarly
Fig. 7 presents the comparison <strong>for</strong> dairy, and the
same assumptions were made here as was the case
<strong>for</strong> the presentation <strong>in</strong> Fig. 6. Interest<strong>in</strong>gly it is
Turkey that has a higher number of specific regulations
<strong>for</strong> dairy products than the EU.
7
regulations /
country
10
8
6
4
2
0
3
EUROPE
[39 countries]
3
Dairy
EU MS
[25 countries]
2
non-EU MS
[14 countries]
4
Turkey
Figure 7. Average number of mycotox<strong>in</strong> regulations
[2006] <strong>for</strong> dairy (products) per country <strong>in</strong> Europe, European
Union member states, non-EU countries and
Turkey.
6. Current <strong>in</strong>ternational events, European
projects and European developments
The cont<strong>in</strong>uous <strong>in</strong>terest of the scientific community
<strong>for</strong> the developments <strong>in</strong> mycotox<strong>in</strong> regulations
is witnessed by the scientific programmes
of lead<strong>in</strong>g <strong>in</strong>ternational mycotox<strong>in</strong> conferences.
The 4 th World Mycotox<strong>in</strong> Forum, held <strong>in</strong> C<strong>in</strong>c<strong>in</strong>nati,
November 2006, started with a plenary session
where attention was given on “recent regulatory
developments and on the economic impact
of regulations on <strong>in</strong>ternational trade” (see
www.bastiaanse-communication.com). The XII th
IUPAC International Symposium on <strong>Mycotox<strong>in</strong>s</strong>
and Phycotox<strong>in</strong>s, which will be held <strong>in</strong> Istanbul,
Turkey, May 2007, <strong>in</strong>cludes a topic area:
“Risk assessment, regulations and <strong>in</strong>ternational
trade” (see www.atal.tubitak.gov.tr/iupac200mycotox<strong>in</strong>).
The IUPAC conference will <strong>in</strong>clude a
full-day Turkish national symposium, which will
be an asset <strong>for</strong> Turkish scientists.
Apart from the guid<strong>in</strong>g or support<strong>in</strong>g role of
<strong>in</strong>ternational and European organisations <strong>in</strong> the
establishment of mycotox<strong>in</strong> regulations <strong>in</strong> the EU,
there are several EC funded research projects
of which the results may have <strong>in</strong>fluence as well.
Without <strong>in</strong>tentionally ignor<strong>in</strong>g other EC projects,
examples of projects with significant potential
impact <strong>in</strong>clude “BioCop” and “MON ˙ IQA”, two
projects funded <strong>in</strong> the EC’s 6 th Framework Programme.
“BioCop” is an Integrated Project (IP) with
more than 30 partners, focussed on “New Technologies
to Screen Multiple Chemical Contam<strong>in</strong>ants
<strong>in</strong> <strong>Food</strong>s”. The project runs from 2005 until

Hans P. van Egmond and Marco A. Jonker
Table 1
Unique Chemical Abstracts Service registration numbers of mycotox<strong>in</strong>s, regulated <strong>in</strong> the EU and Turkey.
Mycotox<strong>in</strong> Abbrevation used here CAS Registry Number
aflatox<strong>in</strong> B1 afla B1 1162-65-8
aflatox<strong>in</strong> B2
7220-81-7
aflatox<strong>in</strong> G1
1165-39-5
7241-98-7
aflatox<strong>in</strong> G2
aflatox<strong>in</strong>s B1B2G1G2
afla B1B2G1G2
aflatox<strong>in</strong> M1 afla M1 6795-23-9
deoxynivalenol DON 51481-10-8
fumonis<strong>in</strong> B1
116355-83-0
fumonis<strong>in</strong> B2
116355-84-1
HT-2 tox<strong>in</strong> 26934-87-2
ochratox<strong>in</strong> A 303-47-9
patul<strong>in</strong> 149-29-1
T-2 tox<strong>in</strong> 21259-20-1
zearalenone 17924-92-4
2010 (see www.biocop.org). BioCop methods are
not based on chromatography, but on new biotechnologies
such as transcriptomics, proteomics
and biosensors. These methods must eventually
fulfil recognised per<strong>for</strong>mance criteria, so that they
can be accepted <strong>for</strong> en<strong>for</strong>cement purposes. One
of the work packages of BioCop deals with mycotox<strong>in</strong>s,
and more specifically with the development
of new methods (transcriptomics [identification
of chemical residues through genomic f<strong>in</strong>gerpr<strong>in</strong>ts]
and electrochemical sensors [low cost
portable devices]) to determ<strong>in</strong>e a selection of trichothecenes
[DON, NIV, T-2 and HT-2 tox<strong>in</strong>]. If
successful <strong>in</strong> the first phase, the methods will be
<strong>in</strong>terlaboratory-validated, and standardized by
CEN, possibly <strong>in</strong> the period after the project has
been f<strong>in</strong>alized. In view of the fact that the permitted
use of mycotox<strong>in</strong> methods <strong>in</strong> EU legislation
is based on per<strong>for</strong>mance criteria, the potential of
BioCop-developed methods <strong>for</strong> regulatory use is
significant.
“MON ˙ IQA” is a Network of Excellence (NoE),
at the time of writ<strong>in</strong>g just approved <strong>for</strong> EU fund<strong>in</strong>g
<strong>for</strong> the period 2007-2011 MON ˙ IQA has more
than 60 partners, <strong>in</strong>clud<strong>in</strong>g several Turkish partners.
The full title of the project is “Towards harmonisation
regard<strong>in</strong>g monitor<strong>in</strong>g the hazards <strong>in</strong>
the food supply cha<strong>in</strong>”. MON ˙ IQA is directed to
multidiscipl<strong>in</strong>ary themes, named “clusters”. One
of these clusters is “mycotox<strong>in</strong>s”. Among the key
activities is the development of common strategies
<strong>for</strong> harmonis<strong>in</strong>g and validat<strong>in</strong>g (rapid/novel)
detection methods and technologies. This could
8
possibly <strong>in</strong>clude methodologies such as those developed
<strong>in</strong> BioCop. A specific work package of
MON ˙ IQA will evaluate the economic impact of
implementation of the analytical methodology required
by the new regulations <strong>in</strong> the field of food
quality and safety (e.g. effectiveness, efficiency
and consistency), and to compare the impact of
different options <strong>in</strong> qualitative, quantitative and
monetary terms. Risk-benefit analysis is obviously
gett<strong>in</strong>g more attention.
S<strong>in</strong>ce July 2006, the European Commission’s
Jo<strong>in</strong>t Research Centre/Institute <strong>for</strong> Reference
Materials and Measurements (Geel, Belgium) will
fulfils the role of EU Community Reference Laboratory
(CRL) <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong>. The tasks, duties
and requirements <strong>for</strong> CRLs <strong>for</strong> food and feed,
and <strong>for</strong> animal health are laid down <strong>in</strong> article
32 of Regulation EC no. 882/2004 [27]. The
CRL <strong>for</strong> mycotox<strong>in</strong>s has been created <strong>in</strong> order
to take, among other duties, <strong>in</strong>itiatives and to
co-ord<strong>in</strong>ate activities related to the development,
improvement and application of sample preparation
and methods of analysis <strong>for</strong> the official control
of maximum levels <strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong> food
and feed. Among the tasks mentioned is the function
“to provide technical assistance to the Commission
and, upon its request, to participate <strong>in</strong>
<strong>in</strong>ternational <strong>for</strong>a relat<strong>in</strong>g to the area of competence,
concern<strong>in</strong>g <strong>in</strong> particular the standardization
of analytical methods and their implementation”.
So, it is evident that the l<strong>in</strong>k with CEN
will be important.
The position of CEN with regard to mycotox<strong>in</strong>

<strong>Regulations</strong> <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong> <strong>in</strong> <strong>Food</strong>: <strong>Focus</strong> on the European Union and Turkey
Table 2
Maximum tolerated levels of mycotox<strong>in</strong>s <strong>in</strong> foodstuffs and dairy products <strong>in</strong> the European Union [June
2006].
EU member states: Austria, Belgium, Denmark, F<strong>in</strong>land, France, Germany, Greece, Ireland, Italy, Luxemburg,
The Netherlands, Portugal, Spa<strong>in</strong>, United K<strong>in</strong>gdom
Latest EU member states: Czech Republic, Cyprus, Estonia, Hungary, Latvia, Lithuania, Malta, Poland,
Slovakia, Slovenia are members s<strong>in</strong>ce 1 May 2004;
EU candidate member states: Bulgaria, Romania will jo<strong>in</strong> the EU on 1 January 2007.
Commodity (Sum of) <strong>Mycotox<strong>in</strong>s</strong>(s) Limit Legal Basis Remarks
(µg /kg)
<strong>Food</strong>
groundnuts, nuts and dried fruit afla B1 2 EUb A
and processed products thereof,
<strong>in</strong>tended <strong>for</strong> direct human consumption afla B1B2G1G2 4 id. A
or as an <strong>in</strong>gredient <strong>in</strong> foodstuffs
groundnuts to be subjected to sort<strong>in</strong>g, afla B1 8 id. A
or other physical treatment,
be<strong>for</strong>e human consumption or use afla B1B2G1G2 15 id. A
as an <strong>in</strong>gredient <strong>in</strong> foodstuffs
nuts and dried fruit to be subjected to afla B1 5 id. A
sort<strong>in</strong>g, or other physical treatment,
be<strong>for</strong>e human consumption or use as afla B1B2G1G2 10 id. A
an <strong>in</strong>gredient <strong>in</strong> foodstuffs
cereals (<strong>in</strong>clud<strong>in</strong>g buckwheat, afla B1 2 id. A
Fagopyrum sp.) and processed products
thereof <strong>in</strong>tended <strong>for</strong> direct human afla B1B2G1G2 4 id. A
consumption or use as an <strong>in</strong>gredient
<strong>in</strong> foodstuffs
cereals (<strong>in</strong>clud<strong>in</strong>g buckwheat, afla B1 2 id. A
Fagopyrum sp.), with the exception
of maize, to be subjected to afla B1B2G1G2 4 id. A
sort<strong>in</strong>g, or other physical treatment,
be<strong>for</strong>e human consumption or use
as an <strong>in</strong>gredient <strong>in</strong> foodstuffs
maize to be subjected to sort<strong>in</strong>g, afla B1 5 EUc A
or other physical treatment,
be<strong>for</strong>e human consumption or use afla B1B2G1G2 10 id. A
as an <strong>in</strong>gredient <strong>in</strong> foodstuffs
spices: Capsicum spp. (dried afla B1 5 EUd -
fruits thereof, whole or ground,
<strong>in</strong>clud<strong>in</strong>g chillies, chilli powder, afla B1B2G1G2 10 id. -
cayenne and paprika); Piper spp.
(fruits thereof, <strong>in</strong>clud<strong>in</strong>g white
and black pepper); Myristica fragrans
(nutmeg); Z<strong>in</strong>giber offic<strong>in</strong>ale
(g<strong>in</strong>ger); Curcuma longa
baby foods and processed cereal-based afla B1 0.10 EUg B
foods <strong>for</strong> <strong>in</strong>fants and young children
9

Table 2
Hans P. van Egmond and Marco A. Jonker
Commodity (Sum of) <strong>Mycotox<strong>in</strong>s</strong>(s) Limit Legal Basis Remarks
(µg/kg)
<strong>Food</strong>
dietary foods <strong>for</strong> special medical afla B1 0.10 EUg C
purposes <strong>in</strong>tended specifically
<strong>for</strong> <strong>in</strong>fants
raw cereal gra<strong>in</strong>s (<strong>in</strong>clud<strong>in</strong>g raw ochratox<strong>in</strong> A 5 EUd D
rice and buckwheat)
all products derived from cereals id. 3 id. D
(<strong>in</strong>clud<strong>in</strong>g processed cereal products
and cereal gra<strong>in</strong>s <strong>in</strong>tended <strong>for</strong> direct
human consumption)
dried v<strong>in</strong>e fruit (currants, rais<strong>in</strong>s id. 10 id. D
and sultanas)
baby foods and processed cereal-based id. 0.50 EUg E
foods <strong>for</strong> <strong>in</strong>fants and young children
dietary foods <strong>for</strong> special medical id. 0.50 id. F
purposes <strong>in</strong>tended specifically
<strong>for</strong> <strong>in</strong>fants
fruit juices and fruit nectar, patul<strong>in</strong> 50 EUe G
<strong>in</strong> particular apple juice, and fruit
juice <strong>in</strong>gredients <strong>in</strong> other beverages
concentrated fruit juice after id. 50 id. G
reconstitution as <strong>in</strong>structed
by the manufacturer
spirit dr<strong>in</strong>ks, cider and other id. 50 id. G
fermented dr<strong>in</strong>ks derived from apples
or conta<strong>in</strong><strong>in</strong>g apple juice
solid apple products, <strong>in</strong>clud<strong>in</strong>g id. 25 id. G
apple compote, apple puree
<strong>in</strong>tended <strong>for</strong> direct consumption
apple juice and solid apple products, id. 10 id. G
<strong>in</strong>clud<strong>in</strong>g apple compote and apple
puree, <strong>for</strong> <strong>in</strong>fants and young
children, and labelled and sold
as <strong>in</strong>tended <strong>for</strong> <strong>in</strong>fants and
young children
other baby foods (as def<strong>in</strong>ed id. 10 EUh G
<strong>in</strong> Article 1 of [EUa]) other than
processed cereal-based foods
10

Table 2
<strong>Regulations</strong> <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong> <strong>in</strong> <strong>Food</strong>: <strong>Focus</strong> on the European Union and Turkey
Commodity (Sum of) <strong>Mycotox<strong>in</strong>s</strong>(s) Limit Legal Basis Remarks
(µg/kg)
<strong>Food</strong>
unprocessed cereals other than DON 1250 EUi -
durum wheat, oats and maize
unprocessed durum wheat and oats id. 1750 id. -
unprocessed maize id. 1750 id. H
cereal flour, <strong>in</strong>clud<strong>in</strong>g maize flour, id. 750 id. -
maize grits and maize meal
bread, pastries, biscuits, cereal id. 500 id. -
snacks and breakfast cereals
pasta (dry) id. 750 id. -
processed cereal-based food id. 200 id. -
<strong>for</strong> <strong>in</strong>fants and young children
and baby food
unprocessed cereals other than zearalenone 100 EUi -
maize
unprocessed maize id. 200 id. H
cereal flour except maize flour id. 75 id. -
maize flour, maize meal, maize grits id. 200 id. H
and ref<strong>in</strong>ed maize oil
bread, pastries, biscuits, other id. 50 id. -
cereal snacks and breakfast cereals
maize snacks and maize-based id. 50 id. H
breakfast cereals
processed maize-based foods id. 20 id. -
<strong>for</strong> <strong>in</strong>fants and young children
other processed cereal-based id. 20 id. -
foods <strong>for</strong> <strong>in</strong>fants and young
children and baby food
11

Table 2
Hans P. van Egmond and Marco A. Jonker
Commodity (Sum of) <strong>Mycotox<strong>in</strong>s</strong>(s) Limit Legal Basis Remarks
(µg/kg)
<strong>Food</strong>
unprocessed maize fumonis<strong>in</strong>s 2000 EUi I
maize grits, maize meal id. 1000 id. I
and maize flour
maize-based foods <strong>for</strong> direct id. 400 id. I
consumption with the exception of
maize grits, maize meal and maize
flour and processed maize-based
foods <strong>for</strong> <strong>in</strong>fants and young
children and baby food
processed maize-based foods <strong>for</strong> id. 200 id. I
<strong>in</strong>fants and young children
and baby food
unprocessed cereals and cereal T-2 and HT-2-tox<strong>in</strong> – EUi J
products
Dairy
milk (raw milk, milk <strong>for</strong> the afla M1 0.05 EUb K
manufacture of milk-based
products and heat-treated milk
as def<strong>in</strong>ed by Council Directive
92/46/EEC, as last amended
by Council Directive 94/71/EC)
<strong>in</strong>fant <strong>for</strong>mulae and follow-on id. 0.025 EUg L
<strong>for</strong>mulae, <strong>in</strong>clud<strong>in</strong>g <strong>in</strong>fant milk
and follow-on milk
dietary foods <strong>for</strong> special medical id. 0.025 id. M
purposes <strong>in</strong>tended specifically
<strong>for</strong> <strong>in</strong>fants
12

Table 2
Remarks
<strong>Regulations</strong> <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong> <strong>in</strong> <strong>Food</strong>: <strong>Focus</strong> on the European Union and Turkey
A Per<strong>for</strong>mance criteria <strong>for</strong> methods of analysis are given
B Baby foods and processed cereal-based foods are def<strong>in</strong>ed <strong>in</strong> <strong>for</strong>mer EU-regulations
as <strong>in</strong>dicated <strong>in</strong> EUg; ”<strong>in</strong>fants” shall mean children under the age of twelve months;
”young children” shall mean children aged between 1 and 3 years - The maximum
level <strong>for</strong> baby foods and processed cereal-based foods <strong>for</strong> <strong>in</strong>fants and young children
refer to the dry matter.
C The maximum level <strong>for</strong> dietary foods <strong>for</strong> special medical purposes <strong>in</strong>tended
specifically <strong>for</strong> <strong>in</strong>fants refer 1) <strong>in</strong> the case of milk and milk products,
to the products ready <strong>for</strong> use or 2) <strong>in</strong> the case of products other than
milk and milk products, to the dry matter. The dry matter is determ<strong>in</strong>ed
<strong>in</strong> accordance with <strong>for</strong>mer EU regulations as <strong>in</strong>dicated <strong>in</strong> EUg.
D Per<strong>for</strong>mance criteria <strong>for</strong> methods of analysis are given.
E Baby foods and processed cereal-based foods are def<strong>in</strong>ed <strong>in</strong> <strong>for</strong>mer EU-regulations
as <strong>in</strong>dicated <strong>in</strong> EUg; ”<strong>in</strong>fants” shall mean children under the age of twelve months;
”young children” shall mean children aged between 1 and 3 years - The maximum
level <strong>for</strong> baby foods and processed cereal-based foods <strong>for</strong> <strong>in</strong>fants and young
childeren refer to the dry matter.
F The maximum level <strong>for</strong> dietary foods <strong>for</strong> special medical purposes <strong>in</strong>tended
specifically <strong>for</strong> <strong>in</strong>fants refer 1) <strong>in</strong> the case of milk and milk products,
to the products ready <strong>for</strong> use or 2) <strong>in</strong> the case of products other than milk
and milk products, to the dry matter. The dry matter is determ<strong>in</strong>ed
<strong>in</strong> accordance with <strong>for</strong>mer EU regulations as <strong>in</strong>dicated <strong>in</strong> EUg.
G s<strong>in</strong>ce 1 november 2003; prevention and reduction of patul<strong>in</strong> contam<strong>in</strong>ation.
[see EUf]; per<strong>for</strong>mance criteria <strong>for</strong> methods of analysis are given.
H If no specific level is fixed be<strong>for</strong>e 1 July 2007, the level will apply thereafter.
I If no specific level is fixed be<strong>for</strong>e 1 October 2007, the level will apply thereafter.
J A maximum level will be fixed, if appropiate, be<strong>for</strong>e 1 July 2007.
K Per<strong>for</strong>mance criteria <strong>for</strong> methods of analysis are given.
L Infant <strong>for</strong>mulae and follow-on <strong>for</strong>mulae are def<strong>in</strong>ed <strong>in</strong> <strong>for</strong>mer EU-regulations
as <strong>in</strong>dicated <strong>in</strong> EUg; the maximum level <strong>for</strong> <strong>in</strong>fant <strong>for</strong>mulae and follow-on <strong>for</strong>mulae
refer to the product ready to use.
M The maximum level <strong>for</strong> dietary foods <strong>for</strong> special medical purposes <strong>in</strong>tended specifically
<strong>for</strong> <strong>in</strong>fants refer 1) <strong>in</strong> the case of milk and milk products, to the products ready
<strong>for</strong> use or 2) <strong>in</strong> the case of products other than milk and milk products, to the dry
matter. The dry matter is determ<strong>in</strong>ed <strong>in</strong> accordance with <strong>for</strong>mer EU regulations as
<strong>in</strong>dicated <strong>in</strong> EUg.
13

Hans P. van Egmond and Marco A. Jonker
Table 3
Maximum tolerated levels of mycotox<strong>in</strong>s <strong>in</strong> foodstuffs and dairy products <strong>in</strong> Turkey [June 2006].
TURKEY [TR]: EU candidate member state
Commodity (Sum of) <strong>Mycotox<strong>in</strong>s</strong>(s) Limit Legal Basis Remarks
(µg/kg)
<strong>Food</strong>
nuts, peanuts and other alfa B1 5 TRa N
dried fruits, figs and rais<strong>in</strong>s
and products produced of them afla B1B2G1G2 10 id. O
cereals and cereal products afla B1 2 id. P
afla B1B2G1G2 4 id. P
spices afla B1 5 id. N
afla B1B2G1G2 10 id. O
other risky products afla B1 5 id. P
afla B1B2G1G2 10 id. Q
baby food afla B1 1 id. R
afla B1B2G1G2 2 id. R
raw gra<strong>in</strong> ochratox<strong>in</strong> A 5 id. R
cereal products id. 3 id. -
dried grapes (rais<strong>in</strong>s) id. 10 id. -
apple juices and other dr<strong>in</strong>ks patul<strong>in</strong> 50 id. S
that conta<strong>in</strong> apple juice
Dairy
milk afla M1 0.05 TRa S
milk powder id. 0.5 id. R
cheese id. 0.25 id. S
baby food [milk based] id. 0.05 id. -
14

Table 3
<strong>Regulations</strong> <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong> <strong>in</strong> <strong>Food</strong>: <strong>Focus</strong> on the European Union and Turkey
Remarks
N <strong>in</strong> <strong>for</strong>ce s<strong>in</strong>ce 2002, but exist<strong>in</strong>g s<strong>in</strong>ce 1990 <strong>for</strong> all foods
O <strong>in</strong> <strong>for</strong>ce s<strong>in</strong>ce 2002, but exist<strong>in</strong>g s<strong>in</strong>ce 1997 <strong>for</strong> all foods
P <strong>in</strong> <strong>for</strong>ce s<strong>in</strong>ce 1990
Q <strong>in</strong> <strong>for</strong>ce s<strong>in</strong>ce 1997 <strong>for</strong> all foods
R <strong>in</strong> <strong>for</strong>ce s<strong>in</strong>ce 2002
S <strong>in</strong> <strong>for</strong>ce s<strong>in</strong>ce 1997
REFERENCES
References EU
EUa Commission Directive 2003/13/EC of 10 February 2003 amend<strong>in</strong>g Directive
96/5/EC on processed cereal-based foods and baby foods <strong>for</strong> <strong>in</strong>fants and
young children. OJEU L 41 (2003) 33-36
EUb Commission Regulation No 466/2001 of 8 March 200 sett<strong>in</strong>g maximum levels
<strong>for</strong> certa<strong>in</strong> contam<strong>in</strong>ants <strong>in</strong> foodstuffs - OJEC L 77 [2001] 1-13
EUc Commission Regulation No 2174/2003 of 12 December 2003 as regards
aflatox<strong>in</strong>s amend<strong>in</strong>g Regulation No 466/2001 - OJEC L 326 (2003) 12-15
EUd Commission Regulation No 472/2002 of 12 march 2002 amend<strong>in</strong>g Regulation
No 466/2001 sett<strong>in</strong>g maximum levels <strong>for</strong> certa<strong>in</strong> contam<strong>in</strong>ants <strong>in</strong> foodstuffs
- OJEC L 75 (2002) 18-20
EUe Commission Regulation No 1425/2003 of 11 August 2003 amend<strong>in</strong>g
Regulation No 466/2001 as regards patul<strong>in</strong> - OJEU L 203 (2003) 1-3
EUf Commission Recommendation of 11 August 2003 on the prevention and
reduction of patul<strong>in</strong> contam<strong>in</strong>ation <strong>in</strong> apple juice and apple juice
<strong>in</strong>gredients <strong>in</strong> other beverages. OJEU L 203 (2003) 54-59
EUg Commission Regulation (EC) No 683/2004 of 13 April 2004 amend<strong>in</strong>g
Regulation (EC) No 466/2001 as regards aflatox<strong>in</strong>s and ochratox<strong>in</strong> A
<strong>in</strong> foods <strong>for</strong> <strong>in</strong>fants and young children - OJEU L 106 (2004) 3-5
EUh Commission Regulation No 455/2004 of 11 March 2004 amend<strong>in</strong>g
Regulation (EC) No 466/2001 as regards patul<strong>in</strong> - OJEU L 74 (2004) 11
EUi Commission Regulation (EC) No 856/2005 of 6 June 2005 amend<strong>in</strong>g
Regulation (EC) No 466/2001 as regards Fusarium tox<strong>in</strong>s - OJEU L
143 (2005) 3 - 8
References Turkey
TRa Turkish <strong>Food</strong> Codex Regulation. 1997. Official Gazette (16 November 1997)
No. 23172: 1-198 and the amendments to this Codex Regulation: Official
Gazette (23 September 2002) No. 24885: 29-40.
15

egulations will also be strengthened as a consequence
of the European Commission’s “Mandate
<strong>for</strong> Standardization addressed to CEN <strong>in</strong>
the Field of Methods of Analysis <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong>
<strong>in</strong> <strong>Food</strong>” [26]. The new mandate falls with<strong>in</strong>
the framework of Regulation EC no. 882/2004
[27]. This Regulation provides that sampl<strong>in</strong>g and
analysis methods used <strong>in</strong> the context of official
controls shall comply with relevant Community
rules or if no such rules exist, with <strong>in</strong>ternationally
recognized rules or protocols, <strong>for</strong> example those
that the European Committee <strong>for</strong> Standardization
(CEN) has accepted. The view of the European
Commission on CEN standards is clear:
“The establishment of standardized methods of
analysis is of utmost importance to guarantee a
uni<strong>for</strong>m application and control of the European
legislation <strong>in</strong> all Member States. Standardized
methods of analysis are an <strong>in</strong>dispensable element
<strong>in</strong> guarantee<strong>in</strong>g a high level of food safety”.
In the text of the mandate a statement is
made that the elaboration of the standards should
be undertaken <strong>in</strong> co-operation with the broadest
possible range of <strong>in</strong>terested groups. Twelve of
such (ma<strong>in</strong>ly European) organizations are specifically
mentioned <strong>in</strong> the mandate, which shows
the importance that the European Commission
attaches to a broad support <strong>for</strong> the standardized
methods. In the annex of the mandate the
methods of analysis are specifically mentioned.
These relate to patul<strong>in</strong>, deoxynivalenol, zearalenone,
aflatox<strong>in</strong> B1, ochratox<strong>in</strong> A, and fumonis<strong>in</strong>s
B1 and B2, <strong>in</strong> various foodstuffs. For these
tox<strong>in</strong>/food comb<strong>in</strong>ations methods are needed <strong>in</strong>
view of (com<strong>in</strong>g) regulations; however, currently
standardized methods are not yet available. An
additional item on the list of tasks to be undertaken
is the preparation of a review of updated
and extended per<strong>for</strong>mance criteria <strong>for</strong> methods
of analysis of mycotox<strong>in</strong>s. The aim is to produce
an updated and extended version of the exist<strong>in</strong>g
CEN criteria document [28], which has been the
basis <strong>for</strong> lay<strong>in</strong>g down the per<strong>for</strong>mance criteria <strong>for</strong>
mycotox<strong>in</strong> methods <strong>in</strong> current EU legislation. It
is expected that the new CEN tasks will be completed
<strong>in</strong> 2009.
7. Conclusions
In 2004 there were approximately 100 countries
<strong>in</strong> the world, which had specific regulations
<strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong> food and feed. Harmonisa-
Hans P. van Egmond and Marco A. Jonker
16
tion of tolerance levels is tak<strong>in</strong>g place <strong>in</strong> several
free trade zones. Especially <strong>in</strong> the EU the number
of harmonized regulations <strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong>
food has rapidly grown, and will further grow <strong>in</strong>
the com<strong>in</strong>g years. With<strong>in</strong> one or two years EU
limits will exist <strong>for</strong> some 50 different tox<strong>in</strong>/food
comb<strong>in</strong>ations. This development is scientifically
supported by complementary activities of various
European organizations (e.g. EFSA, SCOOP,
CEN and CRL). In addition, repeat<strong>in</strong>g conferences
(e.g. World Mycotox<strong>in</strong> Forum and IUPAC
Conferences on <strong>Mycotox<strong>in</strong>s</strong> and Phycotox<strong>in</strong>s) and
several large EU-funded European research and
network<strong>in</strong>g projects (e.g. BioCop, MON ˙ IQA)
have an impact as well. In Turkey also detailed
legal limits exist <strong>for</strong> several mycotox<strong>in</strong>/food comb<strong>in</strong>ations.
Legal limits <strong>in</strong> Turkey exist <strong>for</strong> the
aflatox<strong>in</strong>s, ochratox<strong>in</strong> A and patul<strong>in</strong>. Some limits
are the same as those applied <strong>in</strong> the EU. Clear
differences also exist <strong>in</strong> the regulations, as compared
to the EU. This means that several of the
Turkish mycotox<strong>in</strong> regulations would need to be
modified and extended, should the country become
a member of the European Union. Initiatives
to <strong>in</strong>crease Turkish <strong>in</strong>volvement <strong>in</strong> European
research projects and network<strong>in</strong>g activities will
create a good environment to facilitate this harmonization
process.
8. Acknowledgement
The authors wish to thank the <strong>Food</strong> and Agriculture
Organization of the United Nations <strong>for</strong>
permission to make use of material published
<strong>in</strong> 2004 <strong>in</strong> FAO <strong>Food</strong> and Nutrition Paper 81:
“Worldwide regulations <strong>for</strong> mycotox<strong>in</strong>s <strong>in</strong> food
and feed <strong>in</strong> 2003”. Thanks are also due to Ir.
Frans Verstraete (European Commission, Belgium)
and Prof. Dr. Dilek Heperkan (Istanbul
Technical University, Turkey) <strong>for</strong> their critical assessment
of the data presented <strong>in</strong> tables 2 and 3
of this article.
References
[1] FAO <strong>Food</strong> and Nutrition Paper 81, <strong>Food</strong> and
Agriculture Organization of the United Nations,
Rome, Italy (2004).
[2] Fifty-sixth meet<strong>in</strong>g of the Jo<strong>in</strong>t FAO/WHO Expert
Committee on <strong>Food</strong> Additives (JECFA).
FAO <strong>Food</strong> and Nutrition Paper 74, <strong>Food</strong> and
Agriculture Organization of the United Nations,
Rome, Italy (2001).

<strong>Regulations</strong> <strong>for</strong> <strong>Mycotox<strong>in</strong>s</strong> <strong>in</strong> <strong>Food</strong>: <strong>Focus</strong> on the European Union and Turkey
[3] Fifty-sixth report of the Jo<strong>in</strong>t FAO/WHO Expert
Committee on <strong>Food</strong> Additives, WHO Technical
Report Series 906, World Health Organisation,
Geneva, Switzerland (2002).
[4] Anonymus, The EFSA Journal 353, 1 (2006).
[5] Report EUR 17526 EN, Directorate-General <strong>for</strong>
Industry, Office <strong>for</strong> Official Publications of the
European Communities, Luxembourg, (1997).
[6] M. Miraglia and C. Brera, http://europa.
eu.<strong>in</strong>t/comm/food/fs/scoop/<strong>in</strong>dex en.html,
January (2002).
[7] P. Majerus and K. Kapp, http://europa.
eu.<strong>in</strong>t/comm/food/fs/scoop/<strong>in</strong>dex en.html,
March (2002).
[8] M. Gareis, R.C. Schothorst, A. Vidnes,
C. Bergsten, B. Paulsen, C. Brera,
and M. Miraglia, http://europa.eu.<strong>in</strong>t/
comm/food/fs/scoop/task3210.pdf (2003).
[9] FAO <strong>Food</strong> and Nutrition Paper 55, <strong>Food</strong> and
Agriculture Organization of the United Nations,
Rome, Italy, (1993).
[10] Report of the 32nd session of the Codex Committee
of <strong>Food</strong> Additives and Contam<strong>in</strong>ants,
Beij<strong>in</strong>g, People’s Republic of Ch<strong>in</strong>a, <strong>Food</strong> and
Agriculture Organization, Rome, Italy, 20-24
March 2000.
[11] <strong>Food</strong> and Drug Adm<strong>in</strong>istration Investigative
Operations Manual (2002),
http://www.fda.gov/ora/<strong>in</strong>spect ref/iom/
Contents/ch4 TOC.html.
[12] Commission Regulation (EC) No. 401/2006 of 23
February 2006, Official Journal of the European
Union L70, 12 (2006).
[13] M.W. Trucksess, T.B. Whitaker, H.P. van
Egmond, D.M. Wilson, M. Solfrizzo, D. Abramson,
J. Dorner, G.M. Ware, C. Maragos, B. Hald,
M. Sab<strong>in</strong>o, R.M. Eppley, W.M. Hagler, Journal
of AOAC International 86, 1 (2003).
[14] H.J. Jeur<strong>in</strong>g, Meet<strong>in</strong>g the mycotox<strong>in</strong> menace,
Edts. D. Barug, H.P van Egmond, R. López
Garcia, W.A. van Osenbruggen, A. Visconti,
(Wagen<strong>in</strong>gen Academic Publishers, The Netherlands),
153 (2004).
[15] Commission Directive of 13 February 1991, Official
Journal of the European Communities L60,
16 (1991).
[16] R. Bhat, Work<strong>in</strong>g Document Third Jo<strong>in</strong>t
FAO/WHO/UNEP International Conference
on <strong>Mycotox<strong>in</strong>s</strong>, MYC-CONF/99/4a, Tunis,
Tunisia, 3-6 March 1999.
17
[17] L. Lewis, M. Onsongo, H. Njapau, H. Schurz-
Rogers, G. Luber, S. Kieszak, J. Nyamongo, L.
Backer, A.M. Dahiye, A. Misore, K. DeCock,
and C. Rub<strong>in</strong>, Environmental Health Perspectives
113, 1763 (2005).
[18] Y.Y. Gong, K. Cardwell, A. Hounsa, S. Egal,
P.C. Turner, A.J. Hall, and C.P. Wild, British
Medical Journal 325, 20 (2002).
[19] G.S. Shephard, <strong>in</strong> Meet<strong>in</strong>g the mycotox<strong>in</strong> menace,
Edts. D. Barug, H.P. van Egmond, R. López
Garcia, W.A. Van Osenbruggen, A. Visconti,
(Wagen<strong>in</strong>gen Academic Publishers, The Netherlands)
81 (2004).
[20] G.S. Shephard, N.L. Leggott, S. Stockenström,
N.I.M. Somdyala, W.F.O. Marasas, South Afican
Journal of Science 98, 393 (2002).
[21] Commission Regulation (EC) No 683/2004 of
13 April 2004, Official Journal of the European
Union L 106, 3 (2004) [Reference EUg <strong>in</strong> Table
2].
[22] Commission Regulation No 455/2004 of 11
March 2004, Official Journal of the European
Union L 74, 11 (2004). [Reference EUh <strong>in</strong> Table
2].
[23] Commission Regulation (EC) No 856/2004 of
6 June 2005, Official Journal of the European
Union L 143, 3 (2005). [Reference EUi <strong>in</strong> Table
2].
[24] D. Heperkan, Personal communication, (2006).
[25] Commission Regulation (EC) No 776/2006 of
23 May 2006, Official Journal of the European
Union L136, 3 (2004). [Reference EUg <strong>in</strong> Table
1].
[26] Document M383/EN Health & Consumer Protection
Directorate-general. Directorate D- <strong>Food</strong>
Safety: production and distribution cha<strong>in</strong>. D3-
Chemicals, contam<strong>in</strong>ants and pesticides, Brussels,
7 February 2006.
[27] Regulation (EC) No. 882/ 2004 of the European
Parliament and of the Council of 29 April 2004,
Official Journal of the European Communities
L165, 1 (2004).
[28] CEN Report CR 13505, Brussels, Belgium: European
Committee <strong>for</strong> Standardization (CEN)
(1999).