Commission Regulation (EC) No 401/2006

of 23 February 2006

laying down the methods of sampling and analysis for the official control of the levels of mycotoxins in foodstuffs

(Text with EEA relevance)

THE COMMISSION OF THE EUROPEAN COMMUNITIES,

Having regard to the Treaty establishing the European Community,

Having regard to Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 on official controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules(1), in particular Article 11(4),

Whereas:

(1) Commission Regulation (EC) No 466/2001 of 8 March 2001 setting maximum levels for certain contaminants in foodstuffs(2) provides for maximum limits for certain mycotoxins in certain foodstuffs.

(2) Sampling plays a crucial part in the precision of the determination of the levels of mycotoxins, which are very heterogeneously distributed in a lot. It is therefore necessary to fix general criteria which the sampling method should comply with.

(3) It is also necessary to fix general criteria which the method of analysis should comply with in order to ensure that control laboratories use methods of analysis with comparable levels of performance.

(4) Commission Directive 98/53/EC of 16 July 1998 laying down the sampling methods and the methods of analysis for the official control of the levels for certain contaminants in foodstuffs(3) establishes sampling methods and performance criteria for the methods of analysis to be used for the official control of levels of aflatoxins in foodstuffs.

(5) Commission Directive 2002/26/EC of 13 March 2002 laying down the sampling methods and the methods of analysis for the official control of the levels of ochratoxin A in foodstuffs(4), Commission Directive 2003/78/EC of 11 August 2003 laying down the sampling methods and the methods of analysis for the official control of the levels of patulin in foodstuffs(5) and Commission Directive 2005/38/EC of 6 June 2005 laying down the sampling methods and the methods of analysis for the official control of the levels of Fusarium-toxins in foodstuffs(6) similarly establish sampling methods and performance criteria for ochratoxin A, patulin and Fusarium-toxins respectively.

(6) It is appropriate to apply whenever possible the same sampling method to the same product for the control of mycotoxins. Therefore, the sampling methods and performance criteria for the methods of analysis to be used for the official control of all mycotoxins should be brought together in a single legal act in order to make them easier to apply.

(7) Aflatoxins are very heterogeneously distributed in a lot, in particular in a lot of food products with a large particle size such as dried figs or groundnuts. In order to obtain the same representativeness, for batches with food products with large particle size, the weight of the aggregate sample should be larger than in case of batches with food products with a smaller particle size. Since the distribution of mycotoxins in processed products is generally less heterogeneous than in the unprocessed cereal products, it is appropriate to provide for simpler sampling provisions for processed products.

(8) Directives 98/53/EC, 2002/26/EC, 2003/78/EC and 2005/38/EC should therefore be repealed.

(9) It is appropriate that the date of application of this Regulation coincides with the date of application of Commission Regulation (EC) No 856/2005 of 6 June 2005 amending Regulation (EC) No 466/2001 as regards Fusarium toxins(7).

(10) The measures provided for in this Regulation are in accordance with the opinion of the Standing Committee for the Food Chain and Animal Health,

HAS ADOPTED THIS REGULATION:

Article 1U.K.

Sampling for the official control of the levels of mycotoxins in foodstuffs shall be carried out in accordance with the methods set out in Annex I.

Article 2U.K.

Sample preparation and methods of analysis used for the official control of the levels of mycotoxins in foodstuffs shall comply with the criteria set out in Annex II.

Article 3U.K.

Directives 98/53/EC, 2002/26/EC, 2003/78/EC and 2005/38/EC are repealed.

References to the repealed Directives shall be construed as references to this Regulation.

Article 4U.K.

This Regulation shall enter into force on the 20th day following its publication in the Official Journal of the European Union.

It shall apply from 1 July 2006.

F1...

Textual Amendments

ANNEX I(8) U.K.METHODS OF SAMPLING FOR OFFICIAL CONTROL OF THE LEVELS OF MYCOTOXINS IN FOODSTUFFS

A.GENERAL PROVISIONSU.K.

Official controls shall be performed in accordance with the provisions of Regulation (EC) No 882/2004. The following general provisions shall apply without prejudice to the provisions in Regulation (EC) No 882/2004.

A.1.Purpose and scopeU.K.

Samples intended for official control of the levels of mycotoxins content in foodstuffs shall be taken according to the methods set out in this Annex. Aggregate samples thus obtained shall be considered as representative of the lots. Compliance with maximum limits laid down in Regulation (EC) No 466/2001 shall be established on the basis of the levels determined in the laboratory samples.

A.2.DefinitionsU.K.

For the purpose of this Annex, the following definitions shall apply:

A.2.1.

‘lot’ means an identifiable quantity of a food commodity delivered at one time and determined by the official to have common characteristics, such as origin, variety, type of packing, packer, consignor or markings;

A.2.2.

‘sublot’ means a 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;

A.2.3.

‘incremental sample’ means a quantity of material taken from a single place in the lot or sublot;

A.2.4.

‘aggregate sample’ means the combined total of all the incremental samples taken from the lot or sublot;

A.2.5.

‘laboratory sample’ means a sample intended for the laboratory.

A.3.General provisionsU.K.

A.3.1.PersonnelU.K.

Sampling shall be performed by an authorised person F2....

Textual Amendments

A.3.2.Material to be sampledU.K.

Each lot which is to be examined shall be sampled separately. In accordance with the specific sampling provisions for the different mycotoxins, large lots shall be subdivided into sublots to be sampled separately.

A.3.3.Precautions to be takenU.K.

In the course of sampling and preparation of the samples, precautions shall be taken to avoid any changes, which would affect:

Also, all measures necessary to ensure the safety of the persons taking the samples shall be taken.

A.3.4.Incremental samplesU.K.

As far as possible incremental samples shall be taken at various places distributed throughout the lot or sublot. Departure from such procedure shall be recorded in the record provided for under part A.3.8. of this Annex I.

A.3.5.Preparation of the aggregate sampleU.K.

The aggregate sample shall be made up by combining the incremental samples.

A.3.6.Replicate samplesU.K.

The replicate samples for enforcement, trade (defence) and reference (referee) purposes shall be taken from the homogenised aggregate sample, unless such procedure conflicts with [F3the rules applicable in each constituent territory of Great Britain] as regards the rights of the food business operator.

Textual Amendments

A.3.7.Packaging and transmission of samplesU.K.

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.

A.3.8.Sealing and labelling of samplesU.K.

Each sample taken for official use shall be sealed at the place of sampling and identified following the rules [F4applicable in each constituent territory of Great Britain].

Textual Amendments

A record shall 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.

A.4.Different types of lotsU.K.

Food commodities may be traded in bulk, containers, or individual packings, such as sacks, bags, retail packings. The method of sampling may be applied to all the different forms in which the commodities are put on the market.

Without prejudice to the specific provisions set out in other parts of this Annex, the following formula may be used as a guide for the sampling of lots traded in individual packs, such as sacks, bags, retail packings.

B.METHOD OF SAMPLING FOR CEREALS AND CEREAL PRODUCTSU.K.

This method of sampling is of application for the official control of the maximum levels established for aflatoxin B1, total aflatoxins, ochratoxin A and Fusarium-toxins in cereals and cereal products.

B.1.Weight of the incremental sampleU.K.

The weight of the incremental sample shall be about 100 grams, unless otherwise defined in this part B of Annex I.

In the case of lots in retail packings, the weight of the incremental sample shall depend on the weight of the retail pack.

In the case of retail packs of more than 100 grams, this will result in aggregate samples weighing more than 10 kg. If the weight of a single retail pack is much more than 100 grams, then 100 grams shall be taken from each individual retail pack as an incremental sample. This can be done either when the sample is taken or in the laboratory. However, in cases where such method of sampling would lead to unacceptable commercial consequences resulting from damage to the lot (because of packaging forms, means of transport, etc.), then an alternative method of sampling can be applied. For example, in case where a valuable product is marketed in retail packs of 500 grams or 1 kg, the aggregate sample can be obtained by the aggregation of a number of incremental samples that is smaller than the number indicated in Tables 1 and 2, on the condition that the weight of the aggregate sample is equal to the required weight of the aggregate sample mentioned in Tables 1 and 2.

Where the retail pack is less than 100 grams and if the difference is not very large, one retail pack is to be considered as one incremental sample, resulting in an aggregate sample of less than 10 kg. If the weight of the retail pack is much less than 100 grams, one incremental sample consists of two or more retail packs, whereby the 100 grams are approximated as closely as possible.

B.2.General survey of the method of sampling for cereals and cereal productsU.K.

[F5Table 1

Subdivision of lots into sublots depending on product and lot weight

a

Depending on the lot weight — see Table 2.]

Commodity Lot weight (tonne) Weight or number of sublots No incremental samples Aggregate sample Weight (kg)
Cereals and cereal products > 300 and < 1 500 3 sublots 100 10
≥ 50 and ≤ 300 100 tonnes 100 10
< 50 — 3-100 a 1-10

Textual Amendments

B.3.Method of sampling for cereals and cereal products for lots ≥ 50 tonnesU.K.

Textual Amendments

B.4.Method of sampling for cereals and cereal products for lots < 50 tonnesU.K.

For lots of cereals and cereal products less than 50 tonnes, the sampling plan shall be used with 10 to 100 incremental samples, depending on the lot weight, resulting in an aggregate sample of 1 to 10 kg. For very small lots (≤ 0,5 tonnes) a lower number of incremental samples may be taken, but the aggregate sample combining all incremental samples shall be also in that case at least 1 kg.

The figures in Table 2 may be used to determine the number of incremental samples to be taken.

Table 2

Number of incremental samples to be taken depending on the weight of the lot of cereals and cereal products

Lot weight (tonnes)Number of incremental samplesAggregate sample weight(kg)
≤ 0,0531
> 0,05-≤ 0,551
> 0,5-≤ 1101
> 1-≤ 3202
> 3-≤ 10404
> 10-≤ 20606
> 20-≤ 5010010

B.5.Sampling at retail stageU.K.

Sampling of foodstuffs at the retail stage must be done where possible in accordance with the provisions set out in this part B of Annex I.

Where that is not possible, an alternative method of sampling at retail stage may be applied provided that it ensures that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented. In any case, the aggregate sample shall be at least 1 kg(10).

B.6.Acceptance of a lot or sublotU.K.

C.METHOD OF SAMPLING FOR DRIED FRUIT, INCLUDING DRIED VINE FRUIT AND DERIVED PRODUCTS BUT WITH THE EXCEPTION OF DRIED FIGSU.K.

This method of sampling is of application for the official control of the maximum levels established for:

C.1.Weight of the incremental sampleU.K.

The weight of the incremental sample shall be about 100 grams, unless otherwise defined in this part C of Annex I.

In the case of lots in retail packings, the weight of the incremental sample depends on the weight of the retail packing.

In the case of retail packs of more than 100 grams, this will result in aggregate samples weighing more than 10 kg. If the weight of a single retail pack is much more than 100 grams, then 100 grams shall be taken from each individual retail pack as an incremental sample. This can be done either when the sample is taken or in the laboratory. However, in cases where such method of sampling would lead to unacceptable commercial consequences resulting from damage to the lot (because of packaging forms, means of transport, etc.) then an alternative method of sampling can be applied. For example, in case where a valuable product is marketed in retail packs of 500 grams or 1 kg, the aggregate sample can be obtained by the aggregation of a number of incremental samples that is smaller than the number indicated in Tables 1 and 2, on the condition that the weight of the aggregate sample corresponds to the required weight of the aggregate sample mentioned in Tables 1 and 2.

Where the retail pack is less than 100 grams and if the difference is not very large, one retail pack shall be considered as one incremental sample, resulting in an aggregate sample of less than 10 kg. If the weight of the retail pack is much less than 100 grams, one incremental sample shall consist of two or more retail packs, whereby the 100 grams are approximated as closely as possible.

C.2.General survey of the method of sampling dried fruit, with the exception of figsU.K.

Table 1

Subdivision of lots into sublots depending on product and lot weight

a

Depending on the lot weight — see Table 2 of this part of this Annex.

CommodityLot weight (tonnes)Weight or number of sublotsNumber of incremental samplesAggregate sample weight (kg)
Dried fruit≥ 1515-30 tonnes10010
< 15—10-100a1-10

C.3.Method of sampling for dried fruit (lots ≥ 15 tonnes), with the exception of figsU.K.

C.4.Method of sampling for dried fruit (lots < 15 tonnes), with the exception of figsU.K.

For dried fruit lots, with the exception of figs, under 15 tonnes the sampling plan shall be used with 10 to 100 incremental samples, depending on the lot weight, resulting in an aggregate sample of 1 to 10 kg.

The figures in the following table can be used to determine the number of incremental samples to be taken.

Table 2

Number of incremental samples to be taken depending on the weight of the lot of dried fruit

Lot weight (tonnes)Number of incremental samplesAggregate sample weight (kg)
≤ 0,1101
> 0,1-≤ 0,2151,5
> 0,2-≤ 0,5202
> 0,5-≤ 1,0303
> 1,0-≤ 2,0404
> 2,0-≤ 5,0606
> 5,0-≤ 10,0808
> 10,0-≤ 15,010010

C.5.Sampling at retail stageU.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the provisions set out in this part of Annex I.

Where that is not possible, another alternative method of sampling at retail stage may be used provided that it ensures that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented. In any case, the aggregate sample shall be at least 1 kg(11).

C.6.Specific sampling provisions for dried fruit with the exception of dried figs traded in vacuum packsU.K.

For lots equal to or more than 15 tonnes at least 25 incremental samples resulting in a 10 kg aggregate sample shall be taken and for lots less than 15 tonnes, 25 % of the number of incremental samples mentioned in Table 2 shall be taken resulting in an aggregate sample of which the weight corresponds to the weight of the sampled lot (see Table 2).

C.7.Acceptance of a lot or sublotU.K.

D.METHOD OF SAMPLING FOR DRIED FIGS, GROUNDNUTS AND NUTSU.K.

This method of sampling is of application for the official control of the maximum levels established for aflatoxin B1 and total aflatoxins in dried figs, groundnuts and nuts.

[F7D.1. Method of sampling for dried figs U.K.

This method of sampling is of application for the official control of the maximum levels established for aflatoxin B1 and total aflatoxins in dried figs.

D.1.1. Weight of the incremental sample U.K.

The weight of the incremental sample shall be about 300 grams, unless otherwise defined in part D.1 of Annex I.

In the case of lots in retail packings, the weight of the incremental sample depends on the weight of the retail packing.

In the case of retail packs of more than 300 grams, this will result in aggregate samples weighing more than 30 kg. If the weight of a single retail pack is much more than 300 grams, then 300 grams shall be taken from each individual retail pack as an incremental sample. This can be done either when the sample is taken or in the laboratory. However, in cases where such method of sampling would lead to unacceptable commercial consequences resulting from damage to the lot (because of packaging forms, means of transport, etc.), then an alternative method of sampling can be applied. For example, in case where a valuable product is marketed in retail packs of 500 grams or 1 kg, the aggregate sample can be obtained by the aggregation of a number of incremental samples that is smaller than the number indicated in tables 1, 2 and 3, on the condition that the weight of the aggregate sample corresponds to the required weight of the aggregate sample mentioned in tables 1, 2 and 3.

Where the retail pack is less than 300 grams and if the difference is not very large, one retail pack shall be considered as one incremental sample, resulting in an aggregate sample of less than 30 kg. If the weight of the retail pack is much less than 300 grams, one incremental sample shall consist of two or more retail packs, whereby the 300 grams are approximated as closely as possible.

D.1.2. General survey of the method of sampling for dried figs U.K.
Table 1

Subdivision of lots into sublots depending on product and lot weight

a

Depending on the lot weight — see table 2 of this part D.1 of this Annex.

Commodity Lot weight (tonne) Weight or number of sublots No incremental samples Aggregate sample weight (kg)
Dried figs ≥ 15 15-30 tonnes 100 30
< 15 — 10-100 a ≤ 30
D.1.3. Method of sampling for dried figs (lots ≥ 15 tonnes) U.K.
D.1.4. Method of sampling for dried figs (lots < 15 tonnes) U.K.

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.

The figures in the following table 2 may be used to determine the number of incremental samples to be taken and the subsequent division of the aggregate sample.

Table 2

Number of incremental samples to be taken depending on the weight of the lot and number of subdivisions of the aggregate sample

Lot weight (tonnes) No of incremental samples Aggregate sample Weight (kg) (in case of retail packings, weight of aggregate sample can diverge — see point D.1.1) No of laboratory samples from aggregate sample
≤ 0,1 10 3 1 (no division)
> 0,1 – ≤ 0,2 15 4,5 1 (no division)
> 0,2 – ≤ 0,5 20 6 1 (no division)
> 0,5 – ≤ 1,0 30 9 (- < 12 kg) 1 (no division)
> 1,0 – ≤ 2,0 40 12 2
> 2,0 – ≤ 5,0 60 18 (- < 24 kg) 2
> 5,0 – ≤ 10,0 80 24 3
> 10,0 – ≤ 15,0 100 30 3
D.1.5. Method of sampling for derived products and compound foods U.K.
D.1.5.1. Derived products with very small particle weight (homogeneous distribution of aflatoxin contamination) U.K.
D.1.5.2. Other derived products with a relatively large particle size (heterogeneous distribution of aflatoxin contamination) U.K.

Method of sampling and acceptance as for dried figs (D.1.3 and D.1.4).

D.1.6. Sampling at retail stage U.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the provisions set out in this part of Annex I.

Where that is not possible, other effective methods of sampling at retail stage may be used provided that they ensure that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented. In any case, the aggregate sample shall be at least 1 kg (12) .

D.1.7. Specific method of sampling of dried figs and derived products traded in vacuum packs U.K.
D.1.7.1. Dried figs U.K.

For lots equal to or more than 15 tonnes at least 50 incremental samples resulting in a 30 kg aggregate sample shall be taken and for lots of less than 15 tonnes, 50 % of the number of incremental samples mentioned in table 2 shall be taken resulting in an aggregate sample of which the weight corresponds to the weight of the sampled lot (see table 2).

D.1.7.2. Products derived from dried figs with small particle size U.K.

For lots equal to or more than 50 tonnes at least 25 incremental samples resulting in a 10 kg aggregate sample shall be taken and for lots less than 50 tonnes, 25 % of the number of incremental samples mentioned in table 3 shall be taken resulting in an aggregate sample of which the weight corresponds to the weight of the sampled lot (see table 3).

D.1.8. Acceptance of a lot or sublot U.K.

For dried figs subjected to a sorting or other physical treatment:

For dried figs intended for direct human consumption:

In cases where the aggregate sample is 12 kg or less:

Textual Amendments

D.2. Method of sampling for groundnuts (peanuts), other oilseeds, apricot kernels and tree nuts U.K.

This method of sampling is of application for the official control of the maximum levels established for aflatoxin B1 and total aflatoxins in groundnuts (peanuts), other oilseeds, apricot kernels and tree nuts. [F6This method of sampling is of also of application for the official control of the maximum levels established for ochratoxin A, aflatoxin B1 and total aflatoxins in spices with a relatively large particle size (particle size comparable with peanuts or larger e.g. nutmeg).]

D.2.1. Weight of the incremental sample U.K.

The weight of the incremental sample shall be about 200 grams, unless otherwise defined in part D.2 of Annex I.

In the case of lots in retail packings, the weight of the incremental sample depends on the weight of the retail packing.

In the case of retail packs of more than 200 grams, this will result in aggregate samples weighing more than 20 kg. If the weight of a single retail pack is much more than 200 grams, then 200 grams shall be taken from each individual retail pack as an incremental sample. This can be done either when the sample is taken or in the laboratory. However, in cases where such method of sampling would lead to unacceptable commercial consequences resulting from damage to the lot (because of packaging forms, means of transport, etc.), then an alternative method of sampling can be applied. For example, in case where a valuable product is marketed in retail packs of 500 grams or 1 kg, the aggregate sample can be obtained by the aggregation of a number of incremental samples that is smaller than the number indicated in tables 1, 2 and 3, on the condition that the weight of the aggregate sample corresponds to the required weight of the aggregate sample mentioned in tables 1, 2 and 3.

Where the retail pack is less than 200 grams and if the difference is not very large, one retail pack shall be considered as one incremental sample, resulting in an aggregate sample of less than 20 kg. If the weight of the retail pack is much less than 200 grams, one incremental sample shall consist of two or more retail packs, whereby the 200 grams are approximated as closely as possible.

D.2.2. General survey of the method of sampling for groundnuts (peanuts), other oilseeds, apricot kernels and tree nuts U.K.
Table 1

Subdivision of lots into sublots depending on product and lot weight

a

Depending on the lot weight — see table 2 of this part D.2 of this Annex.

Commodity Lot weight (tonne) Weight or number of sublots No incremental samples Aggregate sample weight (kg)
Groundnuts (peanuts), other oilseeds, apricot kernels and tree nuts ≥ 500 100 tonnes 100 20
> 125 and < 500 5 sublots 100 20
≥ 15 and ≤ 125 25 tonnes 100 20
< 15 — 10-100 a ≤ 20
D.2.3. Method of sampling for groundnuts (peanuts), other oilseeds, apricot kernels and tree nuts (lots ≥ 15 tonnes) U.K.
D.2.4. Method of sampling for groundnuts (peanuts), other oilseeds, apricot kernels and tree nuts (lots < 15 tonnes) U.K.

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.

The figures in the following table 2 may be used to determine the number of incremental samples to be taken and the subsequent division of the aggregate sample.

Table 2

Number of incremental samples to be taken depending on the weight of the lot and number of subdivisions of the aggregate sample

Lot weight (tonnes) No of incremental samples Aggregate sample Weight (kg) (in case of retail packings, weight of aggregate sample can diverge — see point D.2.1) No of laboratory samples from aggregate sample
≤ 0,1 10 2 1 (no division)
> 0,1 – ≤ 0,2 15 3 1 (no division)
> 0,2 – ≤ 0,5 20 4 1 (no division)
> 0,5 – ≤ 1,0 30 6 1 (no division)
> 1,0 – ≤ 2,0 40 8 (- < 12 kg) 1 (no division)
> 2,0 – ≤ 5,0 60 12 2
> 5,0 – ≤ 10,0 80 16 2
> 10,0 – ≤ 15,0 100 20 2
D.2.5. Method of sampling for derived products, with the exception of vegetable oil, and compound foods U.K.
D.2.5.1. Derived products (other than vegetable oil) with small particle size, i.e. flour, peanut butter (homogeneous distribution of aflatoxin contamination) U.K.
D.2.5.2. Derived products with a relatively large particle size (heterogeneous distribution of aflatoxin contamination) U.K.

Method of sampling and acceptance as for groundnuts (peanuts), other oilseeds, apricot kernels and tree nuts (D.2.3 and D.2.4).

D.2.6. Sampling at retail stage U.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the provisions set out in this part of Annex I.

Where that is not possible, other effective methods of sampling at retail stage may be used provided that they ensure that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented. In any case, the aggregate sample shall be at least 1 kg (13) .

D.2.7. Specific method of sampling for groundnuts (peanuts), other oilseeds, apricot kernels, tree nuts and derived products traded in vacuum packs U.K.
D.2.7.1. Pistachios, groundnuts (peanuts), Brazil nuts U.K.

For lots equal to or more than 15 tonnes at least 50 incremental samples resulting in a 20 kg aggregate sample shall be taken and for lots of less than 15 tonnes, 50 % of the number of incremental samples mentioned in table 2 shall bé taken resulting in an aggregate sample of which the weight corresponds to the weight of the sampled lot (see table 2).

D.2.7.2. Apricot kernels, tree nuts other than pistachios and Brazil nuts, other oilseeds U.K.

For lots equal to or more than 15 tonnes at least 25 incremental samples resulting in a 20 kg aggregate sample shall be taken and for lots less than 15 tonnes, 25 % of the number of incremental samples mentioned in table 2 shall be taken resulting in an aggregate sample of which the weight corresponds to the weight of the sampled lot (see table 2).

D.2.7.3. Products derived from tree nuts, apricot kernels and groundnuts (peanuts) with small particle size U.K.

For lots equal to or more than 50 tonnes at least 25 incremental samples resulting in a 10 kg aggregate sample shall be taken and for lots less than 50 tonnes, 25 % of the number of incremental samples mentioned in table 3 shall be taken resulting in an aggregate sample of which the weight corresponds to the weight of the sampled lot (see table 3).

D.2.8. Acceptance of a lot or sublot U.K.

For groundnuts (peanuts), other oilseeds, apricot kernels and tree nuts subjected to a sorting or other physical treatment:

For groundnuts (peanuts), other oilseeds, apricot kernels and tree nuts intended for direct human consumption:

In cases where the aggregate sample is 12 kg or less:

E.METHOD OF SAMPLING FOR SPICESU.K.

[F5This method of sampling is of application for the official control of the maximum levels established for ochratoxin A, aflatoxin B1 and total aflatoxins in spices except in cases of spices with a relatively large particle size (heterogeneous distribution of mycotoxin contamination).]

E.1.Weight of the incremental sampleU.K.

The weight of the incremental sample shall be about 100 grams, unless otherwise defined in this part E of Annex I.

In the case of lots in retail packings, the weight of the incremental sample depends on the weight of the retail packing.

In the case of retail packs of >100 grams, this will result in aggregate samples weighing more than 10 kg. If the weight of a single retail pack is >> 100 grams, then 100 grams shall be taken from each individual retail pack as an incremental sample. This can be done either when the sample is taken or in the laboratory. However, in cases where such method of sampling would lead to unacceptable commercial consequences resulting from damage to the lot (because of packaging forms, means of transport, etc.), then an alternative method of sampling can be applied. For example, in case where a valuable product is marketed in retail packs of 500 grams or 1 kg, the aggregate sample can be obtained by the aggregation of a number of incremental samples that is smaller than the number indicated in Tables 1 and 2, on the condition that the weight of the aggregate sample corresponds to the required weight of the aggregate sample mentioned in Tables 1 and 2.

Where the retail pack is less than 100 grams and if the difference is not very large, one retail pack shall be considered as one incremental sample, resulting in an aggregate sample of less than 10 kg. If the weight of the retail pack is much less than 100 grams, one incremental sample shall consist of two or more retail packs, whereby the 100 grams are approximated as closely as possible.

E.2.General survey of the method of sampling for spicesU.K.

Table 1

Subdivision of lots into sublots depending on product and lot weight

a

Depending on the lot weight — see Table 2 of this part of this Annex.

CommodityLot weight (tonnes)Weight or number of sublotsNumber of incremental samplesAggregate sample Weight (kg)
Spices≥ 1525 tonnes10010
< 15—5-100a0,5-10

E.3.Method of sampling for spices (lots ≥ 15 tonnes)U.K.

E.4.Method of sampling for spices (lots < 15 tonnes)U.K.

For lots of spices less than 15 tonnes the sampling plan shall be used with 5 to 100 incremental samples, depending on the lot weight, resulting in an aggregate sample of 0,5 to 10 kg.

The figures in the following Table can be used to determine the number of incremental samples to be taken.

Table 2

Number of incremental samples to be taken depending on the weight of the lot of spices

Lot weight (tonnes)Number of incremental samplesAggregate sample weight (kg)
≤ 0,0150,5
> 0,01-≤ 0,1101
> 0,1-≤ 0,2151,5
> 0,2-≤ 0,5202
> 0,5-≤ 1,0303
> 1,0-≤ 2,0404
> 2,0-≤ 5,0606
> 5,0-≤ 10,0808
> 10,0-≤ 15,010010

E.5.Sampling at retail stageU.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the sampling provisions set out in this part of Annex I.

Where that is not possible, an alternative method of sampling at retail stage may be used provided that it ensures that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented. In any case, the aggregate sample shall be at least 0,5 kg(14).

E.6.Specific method of sampling for spices traded in vacuum packsU.K.

For lots equal to or more than 15 tonnes at least 25 incremental samples resulting in a 10 kg aggregate sample shall be taken and for lots less than 15 tonnes, 25 % of the number of incremental samples mentioned in Table 2 shall be taken resulting in an aggregate sample of which the weight corresponds to the weight of the sampled lot (see Table 2).

E.7.Acceptance of a lot or sublotU.K.

F.METHOD OF SAMPLING FOR MILK AND MILK PRODUCTS; INFANT FORMULAE AND FOLLOW-ON FORMULAE, INCLUDING INFANT MILK AND FOLLOW-ON MILKU.K.

This method of sampling is of application for the official control of the maximum levels established for aflatoxin M1 in milk and milk products and infant formulae and follow-on formulae, including infant milk and follow-on milk and dietary foods (milk and milk products) for special medical purposes intended specifically for infants.

F.1.Method of sampling for milk, milk products, infant formulae and follow-on formulae, including infant milk and follow-on milk.U.K.

The aggregate sample shall be at least 1 kg or 1 litre except where it is not possible e.g. when the sample consists of one bottle.

The minimum number of incremental samples to be taken from the lot shall be as given in Table 1. The number of incremental samples determined is function of the usual form in which the products concerned are commercialised. In the case of bulk liquid products the lot shall be thoroughly mixed insofar as possible and insofar it does not affect the quality of the product, by either manual or mechanical means immediately prior to sampling. In this case, a homogeneous distribution of aflatoxin M1 is assumed within a given lot. It is therefore sufficient to take three incremental samples from a lot to form the aggregate sample.

The incremental samples, which might frequently be a bottle or a package, shall be of similar weight. The weight of an incremental sample shall be at least 100 grams, resulting in an aggregate sample of at least about 1 kg or 1 litre. Departure from this method shall be recorded in the record provided for under part A.3.8 of Annex I.

Table 1

Minimum number of incremental samples to be taken from the lot

Form of commercialisationVolume or weight of lot (in litre or kg)Minimum number of incremental samples to be takenMinimum volume or weight of aggregate sample (in litre or kg)
Bulk—3-51
Bottles/packages≤ 5031
Bottles/packages50 to 50051
Bottles/packages> 500101

F.2.Sampling at retail stageU.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the provisions set out in this part of Annex I.

Where that is not possible, an alternative method of sampling at retail stage may be used provided that it ensures that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented(15).

F.3.Acceptance of a lot or sublotU.K.

[F7G. METHOD OF SAMPLING FOR COFFEE, COFFEE PRODUCTS, LIQUORICE ROOT AND LIQUORICE EXTRACT U.K.

This method of sampling is of application for the official control of the maximum levels established for ochratoxin A in roasted coffee beans, ground roasted coffee, soluble coffee, liquorice root and liquorice extract.

G.1. Weight of the incremental sample U.K.

The weight of the incremental sample shall be about 100 grams, unless otherwise defined in this part G of Annex I.

In the case of lots in retail packings, the weight of the incremental sample shall depend on the weight of the retail packing.

In the case of retail packs of more than 100 grams, this will result in aggregate samples weighing more than 10 kg. If the weight of a single retail pack is much more than 100 grams, then 100 grams shall be taken from each individual retail pack as an incremental sample. This can be done either when the sample is taken or in the laboratory. However, in cases where such method of sampling would lead to unacceptable commercial consequences resulting from damage to the lot (because of packaging forms, means of transport, etc.), then an alternative method of sampling can be applied. For example, in case where a valuable product is marketed in retail packs of 500 grams or 1 kg, the aggregate sample can be obtained by the aggregation of a number of incremental samples that is smaller than the number indicated in tables 1 and 2, on the condition that the weight of the aggregate sample corresponds to the required weight of the aggregate sample mentioned in tables 1 and 2.

Where the retail pack is less than 100 grams and if the difference is not very large, one retail pack shall be considered as one incremental sample, resulting in an aggregate sample of less than 10 kg. If the weight of the retail pack is much less than 100 grams, one incremental sample shall consist of two or more retail packs, whereby the 100 grams are approximated as closely as possible.

G.2. General survey of the method of sampling for roasted coffee, ground roasted coffee, soluble coffee, liquorice root and liquorice extract U.K.

Table 1

Subdivision of lots into sublots depending on product and lot weight

a

Depending on the lot weight — see table 2 of this part of this Annex.

Commodity Lot weight (ton) Weight or number of sublots No incremental samples Aggregate sample Weight (kg)
Roasted coffee beans, ground roasted coffee, soluble coffee, liquorice root and liquorice extract ≥ 15 15-30 tonnes 100 10
< 15 — 10-100 a 1-10

G.3. Method of sampling for roasted coffee beans, ground roasted coffee, soluble coffee liquorice root and liquorice extract (lots ≥ 15 tonnes) U.K.

G.4. Method of sampling for roasted coffee beans, ground roasted coffee, soluble coffee liquorice root and liquorice extract (lots < 15 tonnes) U.K.

For roasted coffee beans, ground roasted coffee, soluble coffee, liquorice root and liquorice extract under 15 tonnes the sampling plan shall be used with 10 to 100 incremental samples, depending on the lot weight, resulting in an aggregate sample of 1 to 10 kg.

The figures in the following table can be used to determine the number of incremental samples to be taken.

Table 2

Number of incremental samples to be taken depending on the weight of the lot of roasted coffee beans, ground roasted coffee, soluble coffee, liquorice root and liquorice extract

Lot weight (tonnes) No of incremental samples Aggregate sample weight (kg)
≤ 0,1 10 1
> 0,1 – ≤ 0,2 15 1,5
> 0,2 – ≤ 0,5 20 2
> 0,5 – ≤ 1,0 30 3
> 1,0 – ≤ 2,0 40 4
> 2,0 – ≤ 5,0 60 6
> 5,0 – ≤ 10,0 80 8
> 10,0 – ≤ 15,0 100 10

G.5. Method of sampling for roasted coffee beans, ground roasted coffee, soluble coffee, liquorice root and liquorice extract traded in vacuum packs U.K.

For lots equal to or more than 15 tonnes at least 25 incremental samples resulting in a 10 kg aggregate sample shall be taken and for lots less than 15 tonnes, 25 % of the number of incremental samples mentioned in table 2 shall be taken resulting in an aggregate sample of which the weight corresponds to the weight of the sampled lot (see table 2).

G.6. Sampling at retail stage U.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the sampling provisions set out in this part of Annex I.

Where that is not possible, an alternative method of sampling at retail stage may be used provided that it ensures that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented. In any case, the aggregate sample shall be at least 1 kg (16) .

G.7. Acceptance of a lot or sublot U.K.

H.METHOD OF SAMPLING FOR FRUIT JUICES INCLUDING GRAPE JUICE, GRAPE MUST, CIDER AND WINEU.K.

This method of sampling is of application for the official control of the maximum levels established for

H.1.Method of samplingU.K.

The aggregate sample shall be at least one litre except where it is not possible e.g. when the sample consists of one bottle.

The minimum number of incremental samples to be taken from the lot shall be as given in Table 1. The number of incremental samples determined is function of the usual form in which the products concerned are commercialised. In the case of bulk liquid products the lot shall be thoroughly mixed insofar as possible and insofar it does not affect the quality of the product, by either manual or mechanical means immediately prior to sampling. In this case, a homogeneous distribution of ochratoxin A and patulin can be assumed within a given lot. It is therefore sufficient to take three incremental samples from a lot to form the aggregate sample.

The incremental samples, which might frequently be a bottle or a package, shall be of similar weight. The weight of an incremental sample shall be at least 100 grams, resulting in an aggregate sample of at least about 1 litre. Departure from this method shall be recorded in the record provided for under part A.3.8 of Annex I.

Table 1

Minimum number of incremental samples to be taken from the lot

Form of commercialisationVolume of lot (in litres)Minimum number of incremental samples to be takenMinimum volume of the aggregate sample (in litres)
Bulk (fruit juice, spirit drinks, cider, wine)—31
Bottles/packages (fruit juice, spirit drinks, cider)≤ 5031
Bottles/packages (fruit juice, spirit drinks, cider)50 to 50051
Bottles/packages (fruit juice, spirit drinks, cider)> 500101
Bottles/packages wine≤ 5011
Bottles/packages wine50 to 50021
Bottles/packages wine> 50031

H.2.Sampling at retail stageU.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the provisions set out in this part of Annex I(17).

Where that is not possible, an alternative method of sampling at retail stage may be used provided that it ensures that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented.

H.3.Acceptance of a lot or sublotU.K.

[F5I. METHOD OF SAMPLING FOR SOLID APPLE PRODUCTS U.K.

This method of sampling is of application for the official control of the maximum levels established for patulin in solid apple products, including solid apple products for infants and young children.]

I.1.Method of samplingU.K.

The aggregate sample shall be at least 1 kg, except where it is not possible e.g. when sampling a single package.

The minimum number of incremental samples to be taken from the lot shall be as given in Table 1. [F8In the case of liquid products the lot shall be thoroughly mixed insofar as possible by either manual or mechanical means immediately prior to sampling. In this case, a homogeneous distribution of patulin can be assumed within a given lot. It is therefore sufficient to take three incremental samples from a lot to form the aggregate sample.]

Textual Amendments

The incremental samples shall be of similar weight. The weight of an incremental sample shall be at least 100 grams, resulting in an aggregate sample of at least 1 kg. Departure from this method shall be recorded in the record provided for under part A.3.8 of Annex I.

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 takenAggregate sample weight(kg)
< 5031
50 to 50051
> 500101

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.

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 lotNumber of packages or units to be takenAggregate sample weight(kg)
1 to 251 package or unit1
26 to 100about 5 %, at least two packages or units1
> 100about 5 %, at maximum 10 packages or units1

I.2.Sampling at retail stageU.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the sampling provisions set out in this part of the Annex.

Where that is not possible, an alternative method of sampling at retail stage may be used provided that it ensures that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented(18).

I.3.Acceptance of a lot or sublotU.K.

J.METHOD OF SAMPLING FOR BABY FOODS AND PROCESSED CEREAL BASED FOODS FOR INFANTS AND YOUNG CHILDRENU.K.

This method of sampling is of application for the official control of the maximum levels established:

J.1.Method of samplingU.K.

J.2.Sampling at retail stageU.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the provisions set out in this part of Annex I.

Where that is not possible, an alternative method of sampling at retail stage may be used provided that it ensures that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented(19).

J.3.Acceptance of a lot or sublotU.K.

[F9K. METHOD OF SAMPLING FOR VEGETABLE OILS U.K.

This method of sampling is of application for the official control of the maximum levels established for mycotoxins, in particular aflatoxin B1, aflatoxin total and zearalenone, in vegetable oils.

K.1. Method of sampling for vegetable oils U.K.

K.2. Method of sampling for vegetable oils at retail stage U.K.

Sampling of foodstuffs at the retail stage shall be done where possible in accordance with the provisions set out in this part of Annex I.

Where that is not possible, other effective methods of sampling at retail stage may be used provided that they ensure that the aggregate sample is sufficiently representative of the sampled lot and is fully described and documented. In any case, the aggregate sample shall be at least 1 kg (20) .

K.3. Acceptance of a lot or sublot U.K.

Textual Amendments

[F6L. METHOD OF SAMPLING FOR VERY LARGE LOTS OR LOTS STORED OR TRANSPORTED IN A WAY WHEREBY SAMPLING THROUGHOUT THE LOT IS NOT FEASIBLE U.K.

L.1. General principles U.K.

In case the way of transport or storage of a lot does not enable to take incremental samples throughout the whole lot, sampling of such lots should preferably be done when the lot is in flow (dynamic sampling).

In the case of large warehouses destined to store food, operators should be encouraged to install equipment in the warehouse enabling (automatic) sampling across the whole stored lot.

When the sampling procedures as provided for in this part L are applied, the food business operator or his representative should be informed of the sampling procedure. If the sampling procedure is questioned by the food business operator or his representative, the food business operator or his representative shall enable the competent authority to sample throughout the whole lot at his/her own cost.

Sampling of a part of the lot is allowed, on the condition that the quantity of the sampled part is at least 10 % of the lot to be sampled. If a part of a lot of food of the same class or description has been sampled and identified as not satisfying [F10legal] requirements, it shall be presumed that the entire lot is also affected, unless further detailed assessment shows no evidence that the rest of the lot is unsatisfactory.

Textual Amendments

The relevant provisions, such as weight of the incremental sample, provided for in the other parts of this Annex are applicable for the sampling for very large lots or lots stored or transported in a way whereby sampling throughout the lot is not feasible.

L.2. Number of incremental samples to be taken in the case of very large lots U.K.

In the case of large sampled portions (sampled portions > 500 tonnes), the number of incremental samples to be taken = 100 incremental samples + √tonnes. However in case the lot is less than 1 500 tonnes and can be subdivided into sublots in accordance with the table 1 of part B and on the condition that the sublots can be separated physically, the number of incremental samples as provided for in part B have to be taken.

L.3. Large lots transported by ship U.K.

L.3.1. Dynamic sampling of large lots transported by ship U.K.

The sampling of large lots in ships is preferably carried out while the product is in flow (dynamic sampling).

The sampling is to be done per hold (entity that can physically be separated). Holds are however emptied partly one after the other so that the initial physical separation no longer exists after transfer into storage facilities. Sampling can therefore be performed based on initial physical separation or based on the separation after transfer into the storage facilities.

The unloading of a ship can last for several days. Normally, sampling has to be performed at regular intervals during the whole duration of unloading. It is however not always feasible or appropriate for an official inspector to be present for sampling during the whole operation of unloading. Therefore sampling of part of the lot is allowed to be undertaken (sampled portion). The number of incremental samples is determined by taking into account the size of the sampled portion.

Even if the official sample is taken automatically, the presence of an inspector is necessary. However if the automatic sampling is done with pre-set parameters which cannot be changed during the sampling and the incremental samples are collected in a sealed receptacle, preventing any possible fraud, then the presence of an inspector is only required at the beginning of the sampling, every time the receptacle of the samples needs to be changed and at the end of the sampling.

L.3.2. Sampling of lots transported by ship by static sampling U.K.

In cases where the sampling is done in a static way the same procedure as foreseen for storage facilities (silos) accessible from above has to be applied (see point L.5.1).

The sampling has to be performed on the accessible part (from above) of the lot/hold. The number of incremental samples is determined by taking into account the size of the sampled portion.

L.4. Sampling of large lots stored in warehouses U.K.

The sampling has to be performed on the accessible part of the lot. The number of incremental samples is determined by taking into account the size of the sampled portion.

L.5. Sampling of storage facilities (silos) U.K.

L.5.1. Sampling of silos (easily) accessible from above U.K.

The sampling has to be performed on the accessible part of the lot. The number of incremental samples is determined by taking into account the size of the sampled portion.

L.5.2. Sampling of silos not accessible from above (closed silos) U.K.
L.5.2.1. Silos not accessible from above (closed silos) with individual sizes > 100 tonnes U.K.

Food stored in such silos cannot be sampled in a static way. Therefore when the food in the silo has to be sampled and there is no possibility to move the consignment, the agreement has to be made with the operator that he or she has to inform the inspector about when the silo will be unloaded, partially or completely, in order to enable sampling when the food is in flow.

L.5.2.2. Silos not accessible from above (closed silos) with individual sizes < 100 tonnes U.K.

Contrary to the provision in part point L.1 (sampled part at least 10 %), the sampling procedure involves the release into a receptacle of a quantity of 50 to 100 kg and taking the sample from it. The size of the aggregate sample corresponds to the whole lot and the number of incremental samples relate to the quantity of the food from the silo released into the receptacle for sampling.

L.6. Sampling of loose food in large closed containers U.K.

Such lots can often only be sampled when unloaded. In certain cases it is not possible to unload at the point of import or control and therefore the sampling should take place when such containers are unloaded. The operator has to inform the inspector about the place and time of unloading the containers.

M. METHOD OF SAMPLING OF FOOD SUPPLEMENTS BASED ON RICE FERMENTED WITH RED YEAST MONASCUS PURPUREUS U.K.

This method of sampling is applicable to the official control of the maximum level established for citrinin in food supplements based on rice fermented with red yeast Monascus purpureus.

Sampling procedure and sample size U.K.

The sampling procedure is on the supposition that the food supplements based on rice fermented with red yeast Monascus purpureus are marketed in retail packages containing usually 30 to 120 capsules per retail package.

Lot size (number of retail packages) Number of retail packages to be taken for sample Sample size
1-50 1 All capsules
51-250 2 All capsules
251- 1 000 4 From each retail package taken for sample, half of the capsules
> 1 000 4 + 1 retail package per 1 000 retail packages with a maximum of 25 retail packages

≤ 10 retail packages: from each retail package, half of the capsules

> 10 retail packages: from each retail package, an equal number of capsules is taken to result in a sample with the equivalent of the content of retail 5 packages]

ANNEX IIU.K.CRITERIA FOR SAMPLE PREPARATION AND FOR METHODS OF ANALYSIS USED FOR THE OFFICIAL CONTROL OF THE LEVELS OF MYCOTOXINS IN FOODSTUFFS

1.INTRODUCTIONU.K.

1.1.PrecautionsU.K.

As the distribution of mycotoxins is generally non-homogeneous, samples shall be prepared, and especially homogenised, with extreme care.

The complete sample as received by the laboratory shall be homogenized, in case the homogenisation is performed by the laboratory.

For the analysis of aflatoxins, daylight should be excluded as much as possible during the procedure, since aflatoxin gradually breaks down under the influence of ultra-violet light.

1.2.Calculation of proportion of shell/kernel of whole nutsU.K.

The limits fixed for aflatoxins in Regulation (EC) No 466/2001 apply to the edible part. The level of aflatoxins in the edible part can be determined by:

Approximately 100 whole nuts shall be taken at random separately from the lot or shall be put aside from each aggregate sample. The ratio 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 shall be determined for that sample using the approximately 100 nuts that have been set aside.

2.TREATMENT OF THE SAMPLE AS RECEIVED IN THE LABORATORYU.K.

Each laboratory sample shall be finely grinded and mixed thoroughly 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 of the product shall be determined on a part of the homogenised sample, using a method that has been demonstrated to determine accurately the dry matter content.

3.REPLICATE SAMPLESU.K.

The replicate samples for enforcement, trade (defence) and reference (referee) purposes shall be taken from the homogenised material unless such procedure conflicts with [F11the rules applicable in each constituent territory of Great Britain] as regards the rights of the food business operator.

Textual Amendments

4.METHOD OF ANALYSIS TO BE USED BY THE LABORATORY AND LABORATORY CONTROL REQUIREMENTSU.K.

4.1.DefinitionsU.K.

A number of the most commonly used definitions that the laboratory shall be required to use are the following:

r

=

Repeatability, the value below which the absolute difference between two single test results obtained under repeatability conditions, namely 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 = 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 / ) × 100].

R

=

Reproducibility, the value below which the absolute difference between single test results obtained under reproducibility conditions, namely on identical material obtained by operators in 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 / ) × 100].

[F54.2. General requirements U.K.

Confirmatory methods of analysis used for food control purposes shall comply with the provisions of items 1 and 2 of Annex III to Regulation (EC) No 882/2004.

4.3. Specific requirements U.K.

4.3.1. Specific requirements for confirmatory methods U.K.
4.3.1.1. Performance criteria U.K.

It is recommended that fully validated confirmatory methods (i.e. methods validated by collaborative trials for relevant matrices) are used where appropriate and available. Other suitable validated confirmatory methods (e.g. methods validated in-house on relevant matrices belonging to the commodity group of interest) may also be used provided they fulfil the performance criteria set out in the following tables.

Where possible, the validation of in-house validated methods shall include a certified reference material.

(a)

Performance criteria for aflatoxins

Note:

  • Values to apply to both B 1 and sum of B 1 + B 2 + G 1 + G 2

  • If sum of individual aflatoxins B 1 + B 2 + G 1 + G 2 are to be reported, then response of each to the analytical system must be either known or equivalent.

[X1Criterion Concentration Range Recommended Value Maximum permitted Value
Blanks All Negligible —
Recovery — Aflatoxin M 1 0,01-0,05 μg/kg 60 to 120 %
> 0,05 μg/kg 70 to 110 %
Recovery — Aflatoxins B 1 , B 2 , G 1 , G 2 < 1,0 μg/kg 50 to 120 %
1-10 μg/kg 70 to 110 %
> 10 μg/kg 80 to 110 %
Reproducibility RSD R All As derived from Horwitz Equation (*)(**) 2 × value derived from Horwitz Equation (*)(**)
Repeatability RSD r may be calculated as 0,66 times Reproducibility RSD R at the concentration of interest.]
(b)

Performance criteria for ochratoxin A

Level μg/kg Ochratoxin A
RSD r % RSD R % Recovery %
< 1 ≤ 40 ≤ 60 50 to 120
≥ 1 ≤ 20 ≤ 30 70 to 110
(c)

Performance criteria for patulin

Level μg/kg Patulin
RSD r % RSD R % Recovery %
< 20 ≤ 30 ≤ 40 50 to 120
20-50 ≤ 20 ≤ 30 70 to 105
> 50 ≤ 15 ≤ 25 75 to 105
(d)

Performance criteria for deoxynivalenol

Level μg/kg Deoxynivalenol
RSD r % RSD R % Recovery %
> 100-≤ 500 ≤ 20 ≤ 40 60 to 110
> 500 ≤ 20 ≤ 40 70 to 120
(e)

Performance criteria for zearalenone

Level μg/kg Zearalenone
RSD r % RSD R % Recovery %
≤ 50 ≤ 40 ≤ 50 60 to 120
> 50 ≤ 25 ≤ 40 70 to 120
(f)

Performance criteria for Fumonisin B 1 and B 2 individually

Level μg/kg Fumonisin B 1 and B 2 individually
RSD r % RSD R % Recovery %
≤ 500 ≤ 30 ≤ 60 60 to 120
> 500 ≤ 20 ≤ 30 70 to 110
(g)

Performance criteria for T-2 and HT-2 toxin individually

Level μg/kg T-2 and HT-2 toxin individually
RSD r % RSD R % Recovery %
15-250 ≤ 30 ≤ 50 60 to 130
> 250 ≤ 25 ≤ 40 60 to 130
(h)

Performance criteria for citrinin

Level μg/kg Citrinin
RSD r % Recommended RSD R % Maximum allowed RSD R % Recovery %
All 0,66 × RSD R As derived from Horwitz Equation (*) (**) 2 × value derived from Horwitz Equation (*) (**) 70 to 120
(i)

Notes to the performance criteria for the mycotoxins:

  • The detection limits of the methods used are not stated as the precision values given at the concentrations of interest.

  • The precision values are calculated from the Horwitz equation, in particular the original Horwitz equation (for concentrations 1,2 × 10 –7 ≤ C ≤ 0,138) (*) and the modified Horwitz equation (for concentrations C < 1,2 × 10 –7 ) (**).

    (*)

    Horwitz equation for concentrations 1,2 × 10 –7 ≤ C ≤ 0,138:

    RSD R = 2 (1-0.5logC)

    (ref: W. Horwitz, L.R. Kamps, K.W. Boyer, J.Assoc.Off.Analy.Chem.,1980, 63, 1344)

    (**)

    Modified Horwitz equation (*) for concentrations C < 1,2 × 10 –7 :

    RSD R = 22 %

    (ref: M. Thompson, Analyst, 2000, 125, p. 385-386)

    Where:

    • RSD R is the relative standard deviation calculated from results generated under reproducibility conditions [(sR/) × 100]

    • C is the concentration ratio (i.e. 1 = 100g/100g, 0,001 = 1 000 mg/kg)

    This is a generalised precision equation which has been found to be independent of analyte and matrix but solely dependent on concentration for most routine methods of analysis.

Editorial Information

4.3.1.2. ‘ Fitness-for-purpose ’ approach U.K.

For in-house validated methods, as an alternative, a ‘fitness-for-purpose’ approach (21) may be used to assess their suitability for official control. Methods suitable for official control must produce results with a standard measurement uncertainty (u) less than the maximum standard measurement uncertainty calculated using the formula below:

where:

If the analytical method provides results with uncertainty measurements less than the maximum standard uncertainty the method shall be considered being equally suitable to one which meets the performance criteria given in point 4.3.1.1.

Table

Numeric values to be used for α as constant in formula set out in this point, depending on the concentration of interest

C (μg/kg) α
≤ 50 0,2
51-500 0,18
501- 1 000 0,15
1 001 - 10 000 0,12
> 10 000 0,1
4.3.2. Specific requirements for semi-quantitative screening methods U.K.
4.3.2.1. Scope U.K.

The scope applies to bioanalytical methods based on immuno-recognition or receptor binding (such as ELISA, dip-sticks, lateral flow devices, immuno-sensors) and physicochemical methods based on chromatography or direct detection by mass spectrometry (e.g. ambient MS). Other methods (e.g. thin layer chromatography) are not excluded provided the signals generated relate directly to the mycotoxins of interest and allow that the principle described hereunder is applicable.

The specific requirements apply to methods of which the result of the measurement is a numerical value, for example a (relative) response from a dip-stick reader, a signal from LC-MS, etc., and that normal statistics apply.

The requirements do not apply to methods that do not give numerical values (e.g. only a line that is present or absent), which require different validation approaches. Specific requirements for these methods are provided in point 4.3.3.

This document describes procedures for the validation of screening methods by means of an inter-laboratory validation, the verification of the performance of a method validated by means of an inter-laboratory exercise and the single-laboratory validation of a screening method.

4.3.2.2. Terminology U.K.

Screening target concentration (STC): the concentration of interest for detection of the mycotoxin in a sample. When the aim is to test compliance with regulatory limits, the STC is equal to the applicable maximum level. For other purposes or in case no maximum level has been established, the STC is predefined by the laboratory.

Screening method: means method used for selection of those samples with levels of mycotoxins that exceed the screening target concentration (STC), with a given certainty. For the purpose of mycotoxin screening, a certainty of 95 % is considered fit-for-purpose. The result of the screening analysis is either ‘ negative ’ or ‘ suspect ’ . Screening methods shall allow a cost-effective high sample-throughput, thus increasing the chance to discover new incidents with high exposure and health risks to consumers. These methods shall be based on bio-analytical, LC-MS or HPLC methods. Results from samples exceeding the cut-off value shall be verified by a full re-analysis from the original sample by a confirmatory method.

‘ Negative sample ’ means the mycotoxin content in the sample is < STC with a certainty of 95 % (i.e. there is a 5 % chance that samples will be incorrectly reported as negative).

‘ False negative sample ’ means the mycotoxin content in the sample is > STC but it has been identified as negative.

‘ Suspect sample ’ (screen positive) means the sample exceeds the cut-off level (see below) and may contain the mycotoxin at a level higher than the STC. Any suspect result triggers a confirmatory analysis for unambiguous identification and quantification of the mycotoxin.

‘ False suspect sample ’ is a negative sample that has been identified as suspect.

‘ Confirmatory methods ’ means methods that provide full or complementary information enabling the mycotoxin to be identified and quantified unequivocally at the level of interest.

Cut-off level: the response, signal, or concentration, obtained with the screening method, above which the sample is classified as ‘ suspect ’ . The cut-off is determined during the validation and takes the variability of the measurement into account.

Negative control (blank matrix) sample: a sample known to be free (22) of the mycotoxin to be screened for, e.g. by previous determination using a confirmatory method of sufficient sensitivity. If no blank samples can be obtained, then material with the lowest obtainable level might be used as long as the level allows the conclusion that the screening method is fit for purpose.

Positive control sample: sample containing the mycotoxin at the screening target concentration, e.g. a certified reference material, a material of known content (e.g. test material of proficiency tests) or otherwise sufficiently characterised by a confirmatory method. In the absence of any of the above, a blend of samples with different levels of contamination or a spiked sample prepared within laboratory and sufficiently characterised can be used, provided it can be proven that the contamination level has been verified.

4.3.2.3. Validation procedure U.K.

The aim of the validation is to demonstrate the fitness of purpose of the screening method. This is done by determination of the cut-off value and determination of the false negative and false suspect rate. In these two parameters performance characteristics such as sensitivity, selectivity, and precision are embedded.

Screening methods can be validated by inter-laboratory or by single laboratory validation. If inter-laboratory validation data is already available for a certain mycotoxin/matrix/STC combination, a verification of method performance is sufficient in a laboratory implementing the method.

4.3.2.3.1. Initial validation by single laboratory validation U.K.

Mycotoxins:

The validation shall be performed for every individual mycotoxin in the scope. In case of bio-analytical methods that give a combined response for a certain mycotoxin group (e.g. aflatoxins B 1 , B 2 , G 1 & G 2 ; fumonisins B 1 & B 2 ), applicability must be demonstrated and limitations of the test mentioned in the scope of the method. Undesired cross-reactivity (e.g. DON-3-glycoside, 3- or 15-acetyl-DON for immuno-based methods for DON) is not considered to increase the false negative rate of the target mycotoxins, but may increase the false suspect rate. This unwanted increasing will be diminished by confirmatory analysis for unambiguous identification and quantification of the mycotoxins.

Matrices:

An initial validation should be performed for each commodity, or, when the method is known to be applicable to multiple commodities, for each commodity group. In the latter case, one representative and relevant commodity is selected from that group (see table A).

Sample set:

The minimum number of different samples required for validation is 20 homogeneous negative control samples and 20 homogeneous positive control samples that contain the mycotoxin at the STC, analysed under intermediate precision (RSD Ri ) conditions spread over 5 different days. Optionally, additional sets of 20 samples containing the mycotoxin at other levels can be added to the validation set to gain insight to what extent the method can distinguish between different mycotoxin concentrations.

Concentration:

For each STC to be used in routine application, a validation has to be performed.

4.3.2.3.2. Initial validation through collaborative trials U.K.

Validation through collaborative trials shall be done in accordance with an internationally recognised protocol on collaborative trials (e.g. ISO 5725:1994 or the IUPAC International Harmonised Protocol) which requires inclusion of valid data from at least eight different laboratories. Other than that, the only difference compared to single laboratory validations is that the ≥ 20 samples per commodity/level can be evenly divided over the participating laboratories, with a minimum of two samples per laboratory.

4.3.2.4. Determination of cut-off level and rate of false suspected results of blank samples U.K.

The (relative) responses for the negative control and positive control samples are taken as basis for the calculation of the required parameters.

Screening methods with a response proportional with the mycotoxin concentration

For screening methods with a response proportional with the mycotoxin concentration the following applies:

Cut-off = R STC – t-value 0,05 * SD STC

R STC =

mean response of the positive control samples (at STC)

t-value:

one tailed t-value for a rate of false negative results of 5 % (see table B)

SD STC =

standard deviation

Screening methods with a response inversely proportional with the mycotoxin concentration

Similarly, for screening methods with a response inversely proportional with the mycotoxin concentration, the cut-off is determined as:

Cut-off = R STC + t-value 0,05 * SD STC

By using this specific t-value for establishing the cut-off value, the rate of false negative results is by default set at 5 %.

Fitness for purpose assessment

Results from the negative control samples are used to estimate the corresponding rate of false suspect results. The t-value is calculated corresponding to the event that a result of a negative control sample is above the cut off value, thus erroneously classified as suspect.

t-value

=

(cut off – mean blank )/SD blank

for screening methods with a response proportional with the mycotoxin concentration

or

t-value

=

( mean blank – cut off)/SD blank

for screening methods with a response inversely proportional with the mycotoxin concentration

From the obtained t-value, based on the degrees of freedom calculated from the number of experiments, the probability of false suspect samples for a one tailed distribution can either be calculated (e.g.. spread sheet function ‘ TDIST ’ ) or taken from a table for t-distribution.

The corresponding value of the one tailed t-distribution specifies the rate of false suspect results.

This concept is described in detail with an example in Analytical and Bioanalytical Chemistry DOI 10.1007/s00216 -013-6922-1.

4.3.2.5. Extension of the scope of the method U.K.
4.3.2.5.1. Extension of scope to other mycotoxins: U.K.

When new mycotoxins are added to the scope of an existing screening method, a full validation is required to demonstrate the suitability of the method.

4.3.2.5.2. Extension to other commodities: U.K.

If the screening method is known or expected to be applicable to other commodities, the validity to these other commodities shall be verified. As long as the new commodity belongs to a commodity group (see Table A) for which an initial validation has already been performed, a limited additional validation is sufficient. For this, a minimum of 10 homogeneous negative control and 10 homogeneous positive control (at STC) samples shall be analysed under intermediate precision conditions. The positive control samples shall all be above the cut-off value. In case this criterion is not met, a full validation is required.

4.3.2.6. Verification of methods already validated through collaborative trials U.K.

For screening methods that have already been successfully validated through a collaborative laboratory trial, the method performance shall be verified. For this a minimum of 6 negative control and 6 positive control (at STC) samples shall be analysed. The positive control samples shall all be above the cut-off value. In case this criterion is not met, the laboratory has to perform a root-cause analysis to identify why it cannot meet the specification as obtained in the collaborative trial. Only after taking corrective action it shall re-verify the method performance in its laboratory. In case the laboratory is not capable to verify the results from the collaborative trial, it will need to establish its own cut-off in a complete single laboratory validation.

4.3.2.7. Continuous method verification/on-going method validation U.K.

After initial validation, additional validation data are acquired by including at least two positive control samples in each batch of samples screened. One positive control sample is a known sample (e.g. one used during initial validation), the other is a different commodity from the same commodity group (in case only one commodity is analysed, a different sample of that commodity is used instead). Inclusion of a negative control sample is optional. The results obtained for the two positive control samples are added to the existing validation set.

At least once a year the cut-off value is re-established and the validity of the method is re-assessed. The continuous method verification serves several purposes:

4.3.2.8. Validation report U.K.

The validation report shall contain:

Note: The cut-off must have the same number of significant figures as the STC. Numerical values used to calculate the cut-off need at least one more significant figure than the STC. U.K.

Note: The statement on the calculated false suspected rate indicates if the method is fit-for-purpose as it indicates the number of blank (or low level contamination) samples that will be subject to verification. U.K.

Table A

Commodity groups for the validation of screening methods

a

If a buffer is used to stabilise the pH changes in the extraction step, then this commodity group can be merged into one commodity group ‘ High water content ’ .

b

‘ Difficult or unique commodities ’ should only be fully validated if they are frequently analysed. If they are only analysed occasionally, validation may be reduced to just checking the reporting levels using spiked blank extracts.

Commodity groups Commodity categories Typical representative commodities included in the category
High water content Fruit Juices Apple juice, grape juice
Alcoholic beverages Wine, beer, cider
Root and tuber vegetables Fresh ginger
Cereal or fruit based purees Purees intended for infants and small children
High oil content Tree nuts Walnut, hazelnut, chestnut
Oil seeds and products thereof Oilseed rape, sunflower, cotton-seed, soybeans, peanuts, sesame etc.
Oily fruits and products thereof Oils and pastes (e.g. peanut butter, tahina)
High starch and/or protein content and low water and fat content Cereal grain and products thereof Wheat, rye, barley, maize, rice, oats Wholemeal bread, white bread, crackers, breakfast cereals, pasta
Dietary products Dried powders for the preparation of food for infants and small children
High acid content and high water content a Citrus products
‘Difficult or unique commodities’b

Cocoa beans and products thereof, copra and products thereof,

coffee, tea

Spices, liquorice

High sugar low water content Dried fruits Figs, raisins, currants, sultanas
Milk and milk products Milk Cow, goat and buffalo milk
Cheese Cow, goat cheese
Dairy products (e.g. milk powder) Yogurt, cream
Table B

One tailed t-value for a false negative rate of 5 %

Degrees of Freedom Number of replicates t-value (5 %)
10 11 1,812
11 12 1,796
12 13 1,782
13 14 1,771
14 15 1,761
15 16 1,753
16 17 1,746
17 18 1,74
18 19 1,734
19 20 1,729
20 21 1,725
21 22 1,721
22 23 1,717
23 24 1,714
24 25 1,711
25 26 1,708
26 27 1,706
27 28 1,703
28 29 1,701
29 30 1,699
30 31 1,697
40 41 1,684
60 61 1,671
120 121 1,658
∞ ∞ 1,645
4.3.3. Requirements for qualitative screening methods (methods that do not give numerical values) U.K.

The development of validation guidelines for binary test methods is currently subject of various standardization bodies (e.g. AOAC, ISO). Very recently AOAC has drafted a guideline on this matter. This document can be regarded as the current state of the art in its field. Therefore methods that give binary results (e.g. visual inspection of dip-stick tests) should be validated according to this guideline

http://www.aoac.org/imis15_prod/AOAC_Docs/ISPAM/Qual_Chem_Guideline_Final_Approved_031412.pdf

4.4. Estimation of measurement uncertainty, recovery calculation and reporting of results (23) U.K.

4.4.1. Confirmatory methods U.K.

The analytical result must be reported as follows:

(a)

Corrected for recovery, the level of recovery being indicated. The correction for recovery is not necessary in case the recovery rate is between 90-110 %.

(b)

As x +/– U whereby x is the analytical result and U is the expanded measurement uncertainty, using a coverage factor of 2 which gives a level of confidence of approximately 95 %.

For food of animal origin, the taking into account of the measurement uncertainty can also be done by establishing the decision limit (CCα) in accordance with Commission Decision 2002/657/EC (24) (point 3.1.2.5 of Annex I — the case of substances with established permitted limit).

However if the result of the analysis is significantly (> 50 %) lower than the maximum level or much higher than the maximum level (i.e. more than 5 times the maximum level), and on the condition that the appropriate quality procedures are applied and the analysis serves only the purpose of checking compliance with legal provisions, the analytical result might be reported without correction for recovery and the reporting of the recovery rate and measurement uncertainty might be omitted in these cases.

The present interpretation rules of the analytical result in view of acceptance or rejection of the lot apply to the analytical result obtained on the sample for official control. In case of analysis for defence or referee purposes, the national rules apply.

4.4.2. Screening methods U.K.

The result of the screening shall be expressed as compliant or suspected to be non-compliant.

‘ Suspected to be non-compliant ’ means the sample exceeds the cut-off level and may contain the mycotoxin at a level higher than the STC. Any suspect result triggers a confirmatory analysis for unambiguous identification and quantification of the mycotoxin.

‘ Compliant ’ means that the mycotoxin content in the sample is < STC with a certainty of 95 % (i.e. there is a 5 % chance that samples will be incorrectly reported as negative). The analytical result is reported as ‘ < level of STC ’ with the level of STC specified.]

4.5.Laboratory quality standardsU.K.

Laboratory must comply with the provisions of Article 12 of Regulation (EC) No 882/2004 on official controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules(25).

(2)

OJ L 77, 16.3.2001, p. 1. Regulation as last amended by Regulation (EC) No 199/2006 (OJ L 32, 4.2.2006, p. 34).

(3)

OJ L 201, 17.7.1998, p. 93. Directive as last amended by Directive 2004/43/EC (OJ L 113, 20.4.2004, p. 14).

(4)

OJ L 75, 16.3.2002, p. 38. Directive as last amended by Directive 2005/5/EC (OJ L 27, 29.1.2005, p. 38).

(8)

A guidance document for competent authorities for the control of compliance with EU legislation on aflatoxins is available at http://europa.eu.int/comm/food/food/chemicalsafety/contaminants/aflatoxin_guidance_en.pdf. The guidance document provides additional practical information but the information contained in the guidance document is subordinate to the provisions in this Regulation.

(9)

[F5The sampling of such lots shall be performed in accordance with the rules set out in part L. Guidance for sampling large lots shall be provided in a guidance document available on the following website: http://ec.europa.eu/food/food/chemicalsafety/contaminants/guidance-sampling-final.pdf

The application of sampling rules in accordance with EN ISO 24333:2009 or GAFTA Sampling Rules 124, applied by food business operators to ensure compliance with provisions in legislation is equivalent to the sampling rules set out in part L.

For the sampling of lots for Fusarium -toxins, the application of sampling rules in accordance with EN ISO 24333:2009 or GAFTA Sampling Rules 124, applied by food business operators to ensure compliance with provisions in legislation is equivalent to the sampling rules set out in part B]

(10)

In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 kg, the aggregate sample weight might be less than 1 kg.

(11)

In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 kg, the aggregate sample weight might be less than 1 kg.

(12)

[F7In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 kg, the aggregate sample weight might be less than 1 kg.]

(13)

[F7In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 kg, the aggregate sample weight might be less than 1 kg.]

(14)

In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 0,5 kg, the aggregate sample weight might be less than 0,5 kg.

(15)

In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 kg, the aggregate sample weight might be less than 1 kg.

(16)

[F7In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 kg, the aggregate sample weight might be less than 1 kg.]

(17)

In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 litre, the aggregate sample volume might be less than 1 litre.

(18)

In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 kg, the aggregate sample weight might be less than 1 kg.

(19)

In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 kg, the aggregate sample weight might be less than 1 kg.

(20)

[F9In case the portion to be sampled is so small that it is impossible to obtain an aggregate sample of 1 kg, the aggregate sample weight might be less than 1 kg.]

(21)

[F5Ref: M. Thompson and R. Wood, Accred. Qual. Assur., 2006, 10, p. 471-478.]

(22)

[F5Samples are considered free of analyte if the amount present in the sample does not exceed more than 1/5 th of the STC. If the level can be quantified with a confirmatory method, the level must be taken into consideration for the validation assessment.]

(23)

[F5More details on procedures for the estimation of measurement uncertainty and on procedures for assessing recovery can be found in the report ‘ Report on the relationship between analytical results, measurement uncertainty, recovery factors and the provisions of EU food and feed legislation ’ — http://ec.europa.eu/food/food/chemicalsafety/contaminants/report-sampling_analysis_2004_en.pdf]

(24)

[F5Commission Decision 2002/657/EC of 14 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results ( OJ L 221, 17.8.2002, p. 8 ).]

(25)

See also the transitional arrangements provided for in article 18 of Commission Regulation (EC) No 2076/2005 of 5 December 2005 laying down transitional arrangements for the implementation of Regulation (EC) No 853/2004, 854/2004 and 882/2004 of the European Parliament and of the Council and amending Regulations (EC) No 853/2004 and 854/2004 (OJ L 338, 22.12.2005, p. 83).

Textual Amendments