1998L0053 — EN — 10.05.2004 — 003.001 — 1
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►B COMMISSIONDIRECTIVE98/53/EC
of16July1998
layingdownthesamplingmethodsandthemethodsofanalysisfortheofficialcontrolofthelevels
forcertain contaminantsinfoodstuffs
(TextwithEEArelevance)
(OJ L 201, 17.7.1998, p. 93)
Amended by:
Official Journal
No page date
►M1Commission Directive 2002/27/EC of 13 March 2002 L 75 44 16.3.2002
►M2Commission Directive 2003/121/EC of 15 December 2003 L 332 38 19.12.2003
►M3Commission Directive 2004/43/EC of 13 April 2004 L 113 14 20.4.2004
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COMMISSIONDIRECTIVE98/53/EC
of16July1998
layingdownthesamplingmethodsandthemethodsofanalysisfor
the official control of the levels for certain contaminants in food-
stuffs
(TextwithEEArelevance)
THE COMMISSION OF THE EUROPEAN COMMUNITIES,
Having regard to the Treaty establishing the European Community,
Having regard to Council Directive 85/591/EEC of 20 December 1985
concerning the introduction of Community methodsof s ampling andanalysis for the monitoring of foodstuffs intended for human consump-tion (
1), and in particular Article 1 thereof,
Whereas Commission Regulation (EC) No 1525/98 of 16 July 1998,
amending Commission Regulation (EC) No 194/97 setting maximumlevelsfor certain contaminantsin foods tuffs(
2) fixesmaximum limits
for aflatoxins in certain foodstuffs;
WhereasCouncil Directive 93/99/EEC of 29 October 1993 on the
subject of additional measures concerning the official control of food-
stuffs (3) introduces a system of quality standards for laboratories
entrusted by the Member States with the official control of foodstuffs;
Whereas sampling plays a crucial part in the precision of the determi-
nation of the levelsof the aflatoxinswhich are very heterogeneous lydistributed in a lot;
Whereas it seems necessary to fix general criteria which the method of
analysis has to comply with in order to ensure that laboratories, in
charge of the control, use methods of analysis with comparable levelsof performance;
Whereas the provisions for the sampling and methods of analysis have
been drawn up on the basis of present knowledge and they may beadapted to take account of advancesin s cientific and technological
knowledge;
Whereas the methods of sampling used currently by the competent
authoritieslargely differ in the Member States ; whereasthe competentauthoritiesin certain Member Statesare not in a pos ition to apply allthe provisions of this Directive in a short time; whereas it is, therefore,
necessary to provide a suitable period to apply these provisions;
WhereasMember Stateswill have to modify their methodsof s ampling
gradually in order to comply with the provisions laid down in theAnnexesto thisDirective by the time the Directive hasto be applied;whereasit is , therefore, appropriate to examine regularly with theMember States the application of these provisions;
Whereasthe meas uresprovided for in thisDirective are in accordance
with the opinion of the Standing Committee on Foodstuffs,
HAS ADOPTED THIS DIRECTIVE:
Article 1
The Member States shall take all measures necessary to ensure that the
sampling for the official control of the levels of aflatoxins in foodstuffsiscarried out in accordance with the methodsdes cribed in Annex I ofthisDirective.1998L0053 — EN — 10.05.2004 — 003.001 — 2
(1) OJ L 372, 31. 12. 1985, p. 50.
(2) See page 43 of thisOfficial Journal.
(3) OJ L 290, 24. 11. 1993, p. 14.
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Article 2
The Member States shall take all measures necessary to ensure that
sample preparation and methods of analyses used for the official
control of the levelsof aflatoxinsin foods tuffscomply with the criteria
described in Annex II of this Directive.
Article 3
The Member Statess hall, not later than 31 December 2000, bring into
force the laws, regulations or administrative provisions necessary tocomply with the provisions of this Directive. They shall forthwithnotify the Commission thereof.
When Member States adopt these provisions, the provisions shall
contain a reference to this Directive or shall be accompanied by suchreference at the time of their official publication. The procedure for
such reference shall be adopted by Member States.
Article 4
ThisDirective s hall enter into force on the 20
thday following itspubli-
cation in the Official Journal of the European Communities .
This Directive is addressed to the Member States.1998L0053 — EN — 10.05.2004 — 003.001 — 3
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ANNEX I
Methodsof samplingfor official checking control of the levels of aflatoxins
incertainfoodstuffs
1.Purposeandscope
Samplesintended for official checking of the levelsof aflatoxin content
in foodstuffs shall be taken according to the methods described below.
Aggregate samples thus obtained shall be considered as representative
of the lots. Compliance with maximum limits laid down in Commission
Regulation (EC) No 1525/98 shall be established on the basis of the
levels determined in the laboratory samples.
2.Definitions
Lot: an identifiable quantity of a food commodity
delivered at one time and determined by the
official to have common characteristics, such
asorigin, variety, type of packing, packer,
consignor or markings.
Sublot: designated part of a large lot in order to apply
the sampling method on that designated part.
Each sublot must be physically separate and
identifiable.
Incremental sample: a quantity of material taken from a single place
in the lot or sublot.
Aggregate sample: the combined total of all the incremental
samples taken from the lot or sublot.
Laboratory sample: sample intended for the laboratory
( = subsample).
3.Generalprovisions
3.1.Personnel
Sampling shall be performed by an authorised person as specified by
the Member States.
3.2.Material to be sampled
Each lot which is to be examined must be sampled separately. In accor-
dance with the specific provisions in point 5 of this Annex, large lots
should be subdivided into sublots to be sampled separately.
3.3.Precautions to be taken
In the course of sampling and preparation of the laboratory samples
precautionsmus t be taken to avoid any changeswhich would affect
the aflatoxin content, adversely affect the analytical determination or
make the aggregate samples unrepresentative.
3.4.Incremental samples
As far as possible incremental samples should be taken at various
places distributed throughout the lot or sublot. Departure from this
procedure must be recorded in the record provided for in 3.8.
3.5.Preparation of the aggregate sample and the laboratory samples
(subsamples)
The aggregate sample is made up by uniting and sufficiently mixing the
incremental samples. After mixing, the aggregate sample must be
divided into equal subsamples in accordance with the specific provi-
sions of point 5 of this Annex.
The mixing is necessary to ensure that each subsample contains
portionsof the whole lot or s ublot.1998L0053 — EN — 10.05.2004 — 003.001 — 4
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3.6.Replicate samples
The replicate samples for enforcement, trade (defence) and referee
purposes are to be taken from the homogenised laboratory sample,
unless this conflicts with Member States' rules on sampling.
3.7.Packaging and transmission of laboratory samples
Each laboratory sample shall be placed in a clean, inert container
offering adequate protection from contamination and against damage
in transit. All necessary precautions shall be taken to avoid any change
in composition of the laboratory sample which might arise during trans-
portation or storage.
3.8.Sealing and labelling of laboratory samples
Each sample taken for official use shall be sealed at the place of
sampling and identified following the Member State's regulations. A
record must be kept of each sampling, permitting each lot to be identi-
fied unambiguously and giving the date and place of sampling together
with any additional information likely to be of assistance to the analyst.
4.Explanatoryprovisions
4.1.Different types of lots
Food commoditiesmay be traded in bulk, containers , or individual
packings (sacks, bags, retail packings, etc.). The sampling procedure
can be applied to all the different formsin which the commoditiesare
put on the market.
Without prejudice to the specific provisions as laid down in point 5 of
thisAnnex, the following formula can be us ed asa guide for the
sampling of lots traded in individual packings (sacks, bags, retail pack-
ings, etc.):
Sampling frequency (SF) =Weight of the lot /C2weight of the incremental sample
Weight of the aggregate sample /C2weight of individual packing
—Weight: in kg
Sampling frequency (SF): every nth sack or bag from which an incremental
sample must be taken (decimal figures should be rounded to the nearestwhole number).
4.2.Weight of the incremental sample
The weight of the incremental sample should be about 300 grams
unless otherwise defined in point 5 of this Annex and with the excep-
tion of spices in which case the weight of the incremental sample is
about 100 grams. In the case of retail packings, the weight of the incre-
mental sample depends on the weight of the retail packing.
4.3.Number of incremental samples for lots of less than 15 tonnes
The number of incremental samples to be taken depends on the weight
of the lot, with a minimum of 10 and a maximum of 100, unless other-
wise defined in point 5 of this Annex. The figures in the following table
may be used to determine the number of incremental samples to be
taken.
Table1: Number of incremental samples to be taken depending on the
weight of the lot
Lot weight
(tonnes)No of incremental samples
≤0,1 10
> 0,1 -≤0,2 15
> 0,2 -≤0,5 20
> 0,5 -≤1,0 30
> 1,0 -≤2,0 40
> 2,0 -≤5,0 60
> 5,0 -≤10,0 801998L0053 — EN — 10.05.2004 — 003.001 — 5
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Lot weight
(tonnes)No of incremental samples
> 10,0 -≤15,0 100
5.Specificprovisions
5.1.General survey of the sampling procedure for groundnuts, nuts, dried
fruit►M1, spices◄and cereals
Table2: Subdivision of lots into sublots depending on product and lot weight
CommodityLot weight
(tonnes)Weight or number
of sublotsNumber of
incremental
samplesAggregate
sample
Weight (kg)
Dried figsand other
dried fruit≥15 15-30 tonnes100 30
< 15 — 10-100 (1)≤30
Groundnuts, pista-
chios, Brazil nuts and
other nuts≥500 100 tonnes100 30
> 125 and < 500 5 sublots 100 30
≥15 and≤125 25 tonnes100 30
< 15 — 10-100 (1)≤30
Cereals ≥1 500 500 tonnes100 30
> 300 and < 1 500 3 sublots 100 30
≥50 and≤300 100 tonnes100 30
< 50 — 10-100 (1) 1-10
Spices ≥15 25 tonnes100 10
< 15 — 10-100 (1) 1-10
(1) Depending on the lot weight — see point 4.3 or 5.3 of this Annex.
5.2.Groundnuts, pistachios and Brazil nuts
Dried figs
Cereals (lots≥50 tonnes)
Spices
5.2.1. Sampling procedure
— On condition that the sublot can be separated physically, each lot
must be subdivided into sublots following Table 2 at point 5.1.
Taking into account that the weight of the lot isnot alwaysan exact
multiple of the weight of the sublots, the weight of the sublot may
exceed the mentioned weight by a maximum of 20 %,
— each sublot must be sampled separately,
— number of incremental samples: 100. In the case of lots under 15
tonnes, the number of incremental samples to be taken depends on
the weight of the lot, with a minimum of 10 and a maximum of 100
(see point 4.3),
— weight of the aggregate sample = 30 kg which has to be mixed and
to be divided into three equal subsamples of 10 kg before grinding
(this division into three subsamples is not necessary in the case of
groundnuts, nuts, dried fruit and maize intended for further sorting
or other physical treatment, however, this will depend upon the1998L0053 — EN — 10.05.2004 — 003.001 — 6
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availability of equipment which isable to homogenis e a 30 kg
sample). In cases where the aggregate sample weights are under
10 kg, the aggregate sample must not be divided into three subsam-
ples. In the case of spices the aggregate sample weighs not more
than 10 kg and therefore no division in subsamples is necessary,
— laboratory sample: a subsample of 10 kg (each subsample must be
separately ground finely and mixed thoroughly to achieve complete
homogenisation, in accordance with the provisions laid down in
Annex II),
— if it is not possible to carry out the method of sampling described
above because of the commercial consequences resulting from
damage to the lot (because of packaging forms, means of transport,
etc.) an alternative method of sampling may be applied provided
that it is as representative as possible and is fully described and
documented.
5.2.2. Acceptance of a lot or sublot
— For groundnuts, nuts, dried fruit and maize subjected to a sorting or
other physical treatment and spices:
— acceptance if the aggregate sample or the average of the
subsamples conforms to the maximum limit, taking into account
the measurement uncertainty and correction for recovery,
— rejection if the aggregate sample or the average of the subsam-
plesexceedsthe maximum limit beyond reas onable doubt taking
into account the measurement uncertainty and correction for
recovery,
— for groundnuts, nuts, dried fruit and cereals intended for direct
human consumption and cereals, with the exception of maize, to
be subjected to a sorting or other physical treatment:
— acceptance if none of the subsamples exceeds the maximum
limit, taking into account the measurement uncertainty and
correction for recovery,
— rejection if one or more of the subsamples exceeds the
maximum limit beyond reasonable doubt taking into account
the measurement uncertainty and correction for recovery,
— where the aggregate sample is under 10 kg:
— acceptance if the aggregate sample conforms to the
maximum limit, taking into account the measurement uncer-
tainty and correction for recovery,
— rejection if the aggregate sample exceeds the maximum limit
beyond reasonable doubt taking into account analytical
uncertainty and correction for recovery.
5.3.Nuts other than groundnuts, pistachios and Brazil nuts
Dried fruit other than figs
Cereals (lotsunder 50 tonnes )
5.3.1. Sampling procedure
For these products, the sampling procedure laid down in point 5.2.1
may be applied. However, taking into account the low incidence of
contamination for these products and/or the newer forms of packaging
in which products can be traded, simpler sampling methods may be
applied.
For cereal lots under 50 tonnes, a sampling plan consisting of,
depending on the lot weight, 10 to 100 incremental samples each of
100 grams, resulting in an aggregate sample of 1 to 10 kg may be
used. The figures in the following table can be used to determine the
number of incremental samples to be taken.
Table3: Number of incremental samples to be taken depending on the
weight of the lot of cereals
Lot weight
(tonnes)Number of incremental samples
≤11 0
>1-≤32 0
>3-≤10 401998L0053 — EN — 10.05.2004 — 003.001 — 7
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Lot weight
(tonnes)Number of incremental samples
>1 0-≤20 60
>2 0-≤50 100
5.3.2. Acceptance of a lot or sublot
See point 5.2.2.
5.4.Milk
5.4.1. Sampling procedure
Sampling in accordance with Commission Decision 91/180/EEC of 14
February 1991 laying down certain methods of analysis and testing of
raw milk and heat-treated milk (1):
— number of incremental samples: minimum 5,
— weight of aggregate sample: minimum 0,5 kg or litres.
5.4.2. Acceptance of a lot or sublot
— Acceptance if the aggregate sample conforms to the maximum limit,
taking into account the measurement uncertainty and correction for
recovery,
— rejection if the aggregate sample exceeds the maximum limit
beyond reasonable doubt taking into account the measurement
uncertainty and correction for recovery.
5.5.Derived products and compound foods
5.5.1. Milk products
5.5.1.1. Sampling procedure
Sampling in accordance with Commission Directive 87/524/EEC of 6
October 1987 laying down Community methodsof s ampling for
chemical analysis for the monitoring of preserved milk products (2).
Number of incremental samples: minimum 5.
For the other milk products an equivalent method of sampling is used.
5.5.1.2. Acceptance of a lot or sublot
— Acceptance if the aggregate sample conforms to the maximum limit,
taking into account the measurement uncertainty and correction for
recovery,
— rejection if the aggregate sample exceeds the maximum limit
beyond reasonable doubt taking into account the measurement
uncertainty and correction for recovery.
5.5.2. Other derived productswith very s mall particle weight, i.e. flour, fig
paste, peanut butter (homogeneous distribution of aflatoxin contamina-
tion)
5.5.2.1. Sampling procedure
— Number of incremental samples: 100. For lots of under 50 tonnes
the number of incremental samples should be 10 to 100, depending
on the lot weight (see Table 3 at point 5.3.1 of this Annex),
— the weight of the incremental sample should be about 100 grams. In
the case of lots in retail packing, the weight of the incremental
sample depends on the weight of the retail packing,
— weight of aggregate sample = 1-10 kg sufficiently mixed.1998L0053 — EN — 10.05.2004 — 003.001 — 8
(1) OJ L 93, 13. 4. 1991, p. 1.
(2) OJ L 306, 28. 10. 1987, p. 24.▼M2
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5.5.2.2. Number of samples to be taken
— The number of aggregate samples to be taken depends on the lot
weight. The division of large lots into sublots must be done as
defined for cereals ►M1in Table 2 under point 5.1 ◄,
— each sublot must be sampled separately.
5.5.2.3 Acceptance of a lot or sublot
— Acceptance if the aggregate sample conforms to the maximum limit,
taking into account the measurement uncertainty and correction for
recovery,
— rejection if the aggregate sample exceeds the maximum limit
beyond reasonable doubt taking into account the measurement
uncertainty and correction for recovery.
5.6.Other derived products with a relatively large particle size (heteroge-
neous distribution of aflatoxin contamination)
Sampling procedure and acceptance asdefined at points5.2 and 5.3 of
thisAnnex for the raw agricultural product.
5.7.Foods intended for infants and young children
5.7.1. Sampling procedure
The sampling procedure as mentioned for milk and derived products as
well asfor compound food in points5.4, 5.5 and 5.6 applies .
5.7.2. Acceptance of a lot
— Acceptance if the aggregate sample conforms to the maximum limit,
taking into account the measurement uncertainty and correction for
recovery,
— Rejection if the aggregate sample exceeds the maximum limit
beyond reasonable doubt, taking into account the measurement
uncertainty and correction for recovery.
6.Samplingatretail stage
Sampling of foodstuffs at the retail stage should be done where possible
in accordance with the above sampling provisions. Where this is not
possible, other effective sampling procedures at retail stage can be
used provided that they ensure sufficient representativeness for the
sampled lot.1998L0053 — EN — 10.05.2004 — 003.001 — 9
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ANNEX II
Sample preparation and criteria for methods of analysis used in official
checkingofthelevelsofaflatoxinsincertainfoodstuffs
1.Introduction
1.1.Precautions
Daylight should be excluded as much as possible during the procedure,
since aflatoxin gradually breaks down under the influence of ultra-violet
light. As the distribution of aflatoxin is extremely non-homogeneous,
samples should be prepared — and especially homogenised — with
extreme care.
All the material received by the laboratory isto be us ed for the preparation
of test material.
1.2.Calculation of proportion of shell/kernel of whole nuts
The limits fixed for aflatoxins in Commission Regulation (EC) No 1525/98
apply to the edible part.
The level of aflatoxinsin the edible part can be determined by:
— shelling samples of nuts ‘in shell’ and the level of aflatoxins is directly
determined in the edible part,
— homogenise the nuts ‘in shell’ by taking them through the sample
preparation procedure. The sampling and analytical procedure must
estimate the weight of nut kernel in the aggregate sample. The weight
of nut kernel in the aggregate sample is estimated after establishing a
suitable factor for the proportion of nut shell to nut kernel in whole
nuts. This proportion is used to ascertain the amount of kernel in the
bulk sample taken through the sample preparation and analysis proce-
dure. Approximately 100 whole nutsare taken at random s eparately
from the lot or are to be put aside from each aggregate sample. Theratio may, for each laboratory sample, be obtained by weighing the
whole nuts, shelling and re-weighing the shell and kernel portions.
However, the proportion of shell to kernel may be established by the
laboratory from a number of samples and so can be assumed for future
analytical work. But if a particular laboratory sample is found to be in
contravention of any limit, the proportion should be determined for that
sample using the approximately 100 nuts that have been set aside.
2.Treatmentofthesampleasreceived inthelaboratory
Finely grind and thoroughly mix each laboratory sample using a process
that has been demonstrated to achieve complete homogenisation.
In case the maximum level applies to the dry matter, the dry matter content
shall be determined on a part of the homogenised sample, using a proce-
dure that hasbeen demons trated to determine accurately the dry matter
content.
3.Subdivisionofsamplesforenforcementanddefencepurposes
The replicate samples for enforcement, trade (defence) and referee purposes
shall be taken from the homogenised material unless this conflicts with
Member States' rules on sampling.
4.Methodofanalysistobeusedbythelaboratoryandlaboratorycontrol
requirements
4.1.Definitions
A number of the most commonly used definitions that the laboratory will
be required to use are given below:
The most commonly quoted precision parameters are repeatability and
reproducibility.
r = repeatability, the value below which the absolute difference
between two single test results obtained under repeatability
conditions (i. e. same sample, same operator, same apparatus,
same laboratory, and short interval of time) may be expected1998L0053 — EN — 10.05.2004 — 003.001 — 10
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to lie within a specific probability (typically 95 %) and hence
r = 2,8 × sr
sr= Standard deviation, calculated from results generated under
repeatability conditions
RSDr= relative standard deviation, calculated from results generated
under repeatability conditions[(Sr/x) × 100], where x isthe
average of results over all laboratories and samples
R = reproducibility, the value below which the absolute difference
between single test results obtained under reproducibility condi-
tions(i. e. on identical material obtained by operatorsin
different laboratories, using the standardised test method) may
be expected to lie within a certain probability (typically 95 %);
R = 2,8 sR
sR= standard deviation, calculated from results under reproducibility
conditions
RSDR= relative standard deviation calculated from results generated
under reproducibility conditions[(SR/x) × 100]
4.2.General requirements
Methods of analysis used for food control purposes must comply whenever
possible with the provisions of points 1 and 2 of the Annex to Council
Directive 85/591/EEC.
4.3.Specific requirements
Where no specific methods for the determination of aflatoxin levels in
foodstuffs are prescribed at Community level, laboratories may select any
method provided the selected method meets the following criteria:
Criterion Concentration range Recommended valueMaximum permitted
value
BlanksAll Negligible
Recovery — Afla-
toxin M1►M10,01-0,05 µg/kg◄ 60 to 120 %
>0,05►M1µg/kg◄ 70 to 110 %
Recovery — Afla-
toxinsB1,B2,G1,G2< 1,0►M1µg/kg◄ 50 to 120 %
1-10►M1µg/kg◄ 70 to 110 %
>10►M1µg/kg◄ 80 to 110 %
Precision RSDRAll Asderived from
Horwitz equation2 × value derived
from Horwitz equa-
tion
Precision RSDrmay be calculated as0,66 timesprecis ion RSDRat the concentration of interest.
Notes:
— Valuesto apply to both B1and sum of B1+B2+G1+G2,
— if sum of individual aflatoxins B1+B2+G1+G2are to be reported, then
response of each to the analytical system must be either known or
equivalent,
— the detection limits of the methods used are not stated as the precision
valuesare given at the concentrationsof interes t,
— the precision values are calculated from the Horwitz equation, i. e.:
RSDR=2(1-0,5 logC)
where:
— RSDRis the relative standard deviation calculated from results
generated under reproducibility conditions[(SR/x) × 100]
— C isthe concentration ratio (i. e. 1 = 100 g/100 g, 0,001 = 1 000 mg/
kg).
This is a generalised precision equation which has been found to be inde-
pendant of analyte and matrix but solely dependent on concentration for
most routine methods of analysis.1998L0053 — EN — 10.05.2004 — 003.001 — 11
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4.4.Recovery calculation and reporting of results
The analytical result is to be reported corrected or uncorrected for recovery.
The manner of reporting and the level of recovery must be reported. The
analytical result corrected for recovery is used for checking compliance
(see Annex I, points 5.2.2, 5.3.2, 5.4.2, 5.5.1.2 and 5.5.2.3).
The analytical result has to be reported as x +/- U whereby x is the analy-
tical result and U is the expanded measurement uncertainty, using a
coverage factor of 2 which givesa level of confidence of approximately
95 %.
4.5.Laboratory quality standards
Laboratoriesmus t comply with Council Directive 93/99/EEC.1998L0053 — EN — 10.05.2004 — 003.001 — 12
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