Commission Directive 2005/10/EC of 4 February 2005 laying down the sampling methods and the methods of analysis for the official control of the levels of benzo(a)pyrene in foodstuffsText with EEA relevance
Date: 2005-02-04
Institutions: European Commission
Subjects: food inspection, food standard, foodstuff, foodstuffs legislation, research body, toxic substance

8.2.2005

EN

Official Journal of the European Union

L 34/15

COMMISSION DIRECTIVE 2005/10/EC

of 4 February 2005

laying down the sampling methods and the methods of analysis for the official control of the levels
of benzo(a)pyrene in foodstuffs

(Text with EEA relevance)

THE COMMISSION OF THE EUROPEAN COMMUNITIES,

HAS ADOPTED THIS DIRECTIVE:

Having regard to the Treaty
Community,

establishing the European

Having regard to Council Directive 85/591/EEC of 20
December 1985 concerning the introduction of Community
methods of sampling and analysis for the monitoring of food-
stuffs intended for human consumption (1), and in particular
Article 1 thereof,

Whereas:

(1)

(2)

(3)

(4)

Commission Regulation (EC) No 466/2001 of 8 March
2001 setting maximum levels for certain contaminants in
foodstuffs (2)
fixes maximum levels for benzo(a)pyrene
laying down the
reference to measures
and makes
sampling and analysis methods to be used.

Council Directive 93/99/EEC of 29 October 1993 on the
subject of additional measures concerning the official
control of foodstuffs (3)
introduces a system of quality
standards
laboratories entrusted by the Member
for
States with the official control of foodstuffs.

comparable

analysis with
It

It seems necessary to fix general criteria, which the
method of analysis has to comply with in order to
in charge of the control, use
ensure that laboratories,
of
of
methods
performance.
that
analytical
results are reported and interpreted in a
uniform way in order to ensure a harmonised enfor-
cement approach. These interpretation rules are of appli-
cation for the analytical result obtained on the sample for
official control. In case of analysis for defence or referee
purposes, the national rules apply.

levels
importance

also of major

is

The measures provided for
accordance with
Committee on the Food Chain and Animal Health,

in this Directive are in
Standing
the

opinion

the

of

Article 1

the sampling for the official control of

The Member States shall take all measures necessary to ensure
that
the levels of
benzo(a)pyrene in foodstuffs is carried out in accordance with
the methods described in the Annex I to this Directive.

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 levels of benzo(a)pyrene in foodstuffs
comply with the criteria described in the Annex II
to this
Directive.

Article 3

The Member States shall bring into force the laws, regulations
and administrative provisions necessary to comply with this
Directive within 12 months following its publication. They
shall forthwith communicate to the Commission the text of
those provisions
correlation table between those
provisions and this Directive.

and a

When Member States adopt those provisions, they shall contain
a reference to this Directive or shall be accompanied by such a
reference on the occasion of their official publication. Member
States shall determine how such reference is to be made.

Article 4

This Directive shall enter into force on the twentieth day
following that of its publication in the Official
Journal of the
European Union.

This Directive is addressed to the Member States.

Done at Brussels, 4 February 2005.

For the Commission

Markos KYPRIANOU
Member of the Commission

(1) OJ L 372, 31.12.1985, p. 50. Directive as amended by Regulation
(EC) No 1882/2003 of the European Parliament and of the Council
(OJ L 284, 31.10.2003, p. 1).

(2) OJ L 77, 16.3.2001, p. 1. Regulation as last amended by Regulation

(EC) No 208/2005 (See page 3 of this Official Journal).

(3) OJ L 290, 24.11.1993, p. 14. Directive as amended by Regulation

(EC) No 1882/2003.

L 34/16

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Official Journal of the European Union

8.2.2005

ANNEX I

METHODS OF SAMPLING FOR OFFICIAL CONTROL OF THE LEVELS OF BENZO(A)PYRENE IN
FOODSTUFFS

1.

Purpose and Scope

Samples intended for official checking of the levels of benzo(a)pyrene 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 levels laid down in Regulation (EC) No 466/2001 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 having been determined by the official
to have common characteristics, such as origin, variety, type of packing, packer, consignor or markings.

‘Sublot’: designated part of a 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.

3. General provisions

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.

3.3. Precautions to be taken

In the course of sampling and preparation of the samples precautions must be taken to avoid any changes, which
would affect the benzo(a)pyrene 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.

3.5. Preparation of the aggregate sample

The aggregate sample is made up by uniting all incremental samples. This aggregate sample is homogenised in the
laboratory unless this is incompatible with implementation of point 3.6.

3.6. Replicate laboratory samples

Replicate laboratory samples for enforcement, trade (defence) and referee purposes shall be taken from the homo-
genised aggregate sample unless this conflicts with Member States’ rules on sampling.

3.7. Packaging and transmission of samples

Each 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 sample, which
might arise during transportation or storage.

3.8. Sealing and labelling of samples

Each sample taken for official use shall be sealed at the place of sampling and identified following the Member State’s
rules.

A record must be kept of each sampling, permitting each lot to be identified unambiguously and giving the date and
place of sampling together with any additional information likely to be of assistance to the analyst.

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Official Journal of the European Union

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4.

Sampling plans

The sampling method applied shall ensure that the aggregate sample is representative for the lot that is to be
controlled.

4.1. Number of incremental samples

In the case of oils, for which a homogeneous distribution of benzo(a)pyrene can be assumed within a given lot, it is
sufficient to take three incremental samples per lot to form the aggregate sample. Reference to the lot number shall
be given. For olive oil and olive pomace oil further information on sampling is given in Commission Regulation (EC)
No 1989/2003 (1).

For other products, the minimum number of incremental samples to be taken from the lot shall be as given in Table
1. The incremental samples shall be of similar weight, no less than 100g each, resulting in an aggregate sample of no
less than 300g (see point 3.5).

TABLE 1

Minimum number of incremental samples to be taken from the lot

Weight of lot (in kg)

Minimum number of incremental samples to be taken

< 50

50 to 500

> 500

3

5

10

If the lot consists of individual packages, then the number of packages which shall be taken to form the aggregate
sample is given in Table 2.

Number of packages (incremental samples) which shall be taken to form the aggregate sample if the lot
consists of individual packages

Number of packages or units in the lot or sublot

Number of packages or units to be taken

TABLE 2

1 to 25

26 to 100

> 100

1 package or unit

About 5 %, at least 2 packages or units

About 5 %, at maximum 10 packages or units

4.2. Sampling at retail 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.

5. Compliance of the lot or sublot with the specification

The control laboratory shall analyse the laboratory sample for enforcement in duplicate analyses in cases where the
obtained result of the first analysis is less than 20 % below or above the maximum level, and in these cases shall
calculate the mean of the results.

The lot is accepted if the result of the first analysis or, where duplicate analysis is necessary, if the mean does not
exceed the respective maximum level (as laid down in Regulation (EC) No 466/2001) taking into account the
measurement uncertainty and correction for recovery.

The lot is non-compliant with the maximum level (as laid down in Regulation (EC) 466/2001) if the result of the
first analysis or, where duplicate analysis is necessary, if the mean exceeds the maximum level beyond reasonable
doubt taking into account the measurement uncertainty and correction for recovery.

(1) OJ L 295, 13.11.2003, p. 57.

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Official Journal of the European Union

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ANNEX II

SAMPLE PREPARATION AND CRITERIA FOR METHODS OF ANALYSIS USED IN OFFICIAL CHECKING OF
THE LEVELS OF BENZO(A)PYRENE IN FOODSTUFFS

1.

Precautions and general considerations for benzo(a)pyrene in food samples

The basic requirement is to obtain a representative and homogeneous laboratory sample without introducing
secondary contamination.

The analyst should ensure that samples do not become contaminated during sample preparation. Containers
should be rinsed with high purity acetone or hexane (p.A., HLPC grade or equivalent) before use to minimise
the risk of contamination. Wherever possible, apparatus coming into contact with the sample should be made of
inert materials e.g. aluminium, glass or polished stainless steel. Plastics such as polypropylene, PTFE etc. should be
avoided because the analyte can adsorb onto these materials.

All of the sample material received by the laboratory is to be used for the preparation of test material. Only very
finely homogenised samples give reproducible results.

There are many satisfactory specific sample preparation procedures which may be used.

2.

Treatment of the sample as received in the laboratory

Finely grind (where relevant) and mix thoroughly the complete aggregate sample using a process that has been
demonstrated to achieve complete homogenisation.

3.

Subdivision of samples for enforcement and defence purposes

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.

Method of analysis to be used by the laboratory and laboratory control requirements

4.1. Definitions

A number of the most commonly used definitions that the laboratory will be required to use are given below:

r =

sr =

RSDr =

R=

sR =

RSDR =

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 expected to lie within a specific probability (typically 95 %) and
hence r ¼ 2:8 (cid:1) sr.

Standard deviation, calculated from results generated under repeatability conditions.

Relative standard deviation, calculated from results generated under
(cid:1)(cid:2)

(cid:3)

repeatability conditions

sr=x

(cid:1) 100

(cid:4)
.

Reproducibility, the value below which the absolute difference between single test results obtained
under reproducibility conditions (i.e., on identical material obtained by operators in different labora-
tories, using the standardised test method), may be expected to lie within a certain probability
(typically 95 %); R ¼ 2:8 (cid:1) sR.

Standard deviation, calculated from results under reproducibility conditions.

(cid:3)

Relative standard deviation calculated from results generated under reproducibility conditions
(cid:1)(cid:2)

(cid:4)
, where x is the average of results over all laboratories and samples.

(cid:1) 100

sR=x

HORRATr = the observed RSDr divided by the RSDr value estimated from the Horwitz equation (1) using the

assumption r = 0.66R.

HORRATR= the observed RSDR value divided by the RSDR value calculated from the Horwitz equation.

U =

the expanded uncertainty, using a coverage factor of 2 which gives a level of confidence of
approximately 95 %.

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Official Journal of the European Union

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4.2.

General requirements

Methods of analysis used for food control purposes must comply with 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 benzo(a)pyrene in food are prescribed at Community level,
laboratories may select any validated method provided the selected method meets the performance criteria
indicated in the Table. The validation should ideally include a certified reference material.

Performance criteria for methods of analysis for benzo(a)pyrene

TABLE

Parameter

Applicability
Detection limit
Limit of quantification
Precision

Recovery
Specificity

Value/comment

Food specified in Regulation (EC) No …/2005
No more than 0,3 μg/kg
No more than 0,9 μg/kg
HORRATr or HORRATR values of less than 1.5 in the validation collaborative
trial
50 %-120 %
Free from matrix or spectral interferences, verification of positive detection

4.3.1. Performance Criteria — Uncertainty Function Approach

However, an uncertainty approach may also be used to assess the suitability of the method of analysis to be used
by the laboratory. The laboratory may use a method which will produce results within a maximum standard
uncertainty. The maximum standard uncertainty can be calculated using the following formula:

q

Uf ¼

ﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃﬃ
(cid:1)(cid:2)
(cid:4)
(cid:3)
2
2 þ

(cid:2)
0:2C

LOD=2

(cid:3)

where:

Uf

LOD

C

is the maximum standard uncertainty

is the limit of detection of the method

is the concentration of interest

If an analytical method provides results with uncertainty measurements less than the maximum standard uncer-
tainty the method will be equally suitable to one which meets the performance characteristics given in the Table.

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, point 5).

The analyst should note the ‘European Commission Report on the relationship between analytical results, the
measurement of uncertainty, recovery factors and the provisions in EU food legislation’ (2).

The analytical result has to be reported as x +/– U whereby x is the analytical result and U is the measurement
uncertainty.

4.5.

Laboratory quality standards

Laboratories must comply with Directive 93/99/EEC.

4.6. Other considerations for the analysis

P r o f i c i e n c y t e s t i n g
Participation in appropriate proficiency testing schemes which comply with the ‘International Harmonised Protocol
for the Proficiency Testing of (Chemical) Analytical Laboratories’ (3) developed under the auspices of IUPAC/
ISO/AOAC.

I n t e r n a l q u a l i t y c o n t r o l

Laboratories should be able to demonstrate that they have internal quality control procedures in place. Examples of
these are the ‘ISO/AOAC/IUPAC Guidelines on Internal Quality Control in Analytical Chemistry Laboratories’ (4).

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REFERENCES
1. W. Horwitz, ‘Evaluation of Analytical Methods for Regulation of Foods and Drugs’, Anal. Chem., 1982, 54, 67A-76A.

2. European Commission Report on the relationship between analytical results, the measurement of uncertainty, recovery

factors and the provisions in EU food legislation, 2004.

(http://europa.eu.int/comm/food/food/chemicalsafety/contaminants/index_en.htm).

3. ISO/AOAC/IUPAC International Harmonised Protocol for Proficiency Testing of (Chemical) Analytical Laboratories,
Edited by M. Thompson and R. Wood, Pure Appl. Chem., 1993, 65, 2123-2144 (Also published in J. AOAC
International, 1993, 76, 926).

4. ISO/AOAC/IUPAC International Harmonised Guidelines for Internal Quality Control in Analytical Chemistry Labora-

tories, Edited by M. Thompson and R. Wood, Pure Appl. Chem., 1995, 67, 649-666.