Commission Implementing Regulation (EU) 2021/808 of 22 March 2021 on the performance of analytical methods for residues of pharmacologically active substances used in food-producing animals and on the interpretation of results as well as on the methods to be used for sampling and repealing Decisions 2002/657/EC and 98/179/EC (Text with EEA relevance)

Generally, the method extension to additional compounds is only possible for analytes, which are similar structure and characteristic-wise compared to those already included in the analytical method. For such a case, the application of a reduced validation scheme is acceptable. Likewise, no divergence from the method description is allowed.

Calibration curves for the additional substances should be prepared according to the validated procedure. Different batches of matrix materials fortified at different concentration levels (cf. 2.2.1, 2.2.2) should be analysed. Trueness, repeatability and within-laboratory reproducibility/intermediate precision should be within a comparable range to those of the other analytes of the originally validated method and in line with the requirements set in 1.2.2. A calculation of CCβ (screening methods) and CCα (confirmation methods) for the new analytes has to be done.

4.3.   Extensions of methods as regards to matrices/species

The inclusion of new matrices or species in an already validated analytical method shall always be a case-by-case decision based on the knowledge and experiences gained so far with the method and preliminary experiments assessing potential matrix effects and interferences. Generally, this will only be possible for matrices that exhibit similar properties and for non-critical analytes (stability, detectability).

Calibration curves (standard or matrix) should be prepared according to the validated procedure. Different batches of matrix material fortified at different concentration levels (cf. 2.2.1, 2.2.2) should be analysed. Trueness, repeatability and within-laboratory reproducibility/intermediate precision should be within an acceptable range to those of the originally validated method and in line with the requirements set in 1.2.2. Depending on the validation approach, a recalculation of CCβ (screening methods) or CCα (confirmation methods) might be necessary.

If the results are not within an acceptable range compared to the values for the original matrix, an additional full validation will be necessary, in order to determine the matrix/species specific performance parameters.

In cases where MRLs for a specific substance differ for certain matrices, it will most likely be difficult to adapt the method scope to the additional matrix/species and concentration, since in this case two modifications have to be considered. In such cases a full validation is recommended.

ANNEX II

SAMPLING PROCEDURES AND OFFICIAL SAMPLE TREATMENT

1. Sample quantity

The minimum sample quantities shall be defined in the national residue control programme. The minimum sample quantities shall be sufficient to enable the approved laboratories to carry out the analytical procedures necessary to complete the screening and the confirmatory analyses. Specifically for poultry, aquaculture, rabbits, farmed game, reptiles and insects, a sample consists out of one or more animals, depending on the requirements of the analytical methods. For eggs, the sample size is at least 12 eggs, according to the analytical methods used. However, the sample size for ostrich eggs is one egg. In case several substance categories need to be analysed in one sample with different analytical methods, the sample size shall be increased accordingly.

2. Division into sub-samples

Unless technically impossible or not required by national legislation, each sample shall be divided into at least two equivalent sub-samples each allowing the complete analytical procedure. The subdivision can take place at the sampling location or in the laboratory.

3. Traceability

Each sample shall be taken in such way that it is always possible to trace it back to the farm of origin and the batch of animals or the individual animal, where relevant. In particular, for milk, according to the choice of the Member State, the samples can be taken, in either of the following places:

• 1. at the farm from the collection tank;

• 1. at the level of the dairy industry, before the milk has been discharged.

4. Sample containers

Samples shall be collected in suitable containers to maintain sample integrity and traceability. In particular, containers shall prevent substitution, cross-contamination and degradation. The containers shall be officially sealed.

5. Sampling report

A report shall be produced after each sampling procedure.

The inspector collects at least the following data in the sampling report:

• 1. address of the competent authorities;

• 1. name of the inspector or identification code;

• 1. official code number of the sample;

• 1. sampling date;

• 1. name and address of the owner or the person having charge of the animals or the animal products;

• 1. name and address of the animal’s farm of origin (when sampling on farm);

• 1. registration number of the establishment-slaughterhouse number;

• 1. animal or product identification;

• 1. animal species;

• 1. sample matrix;

• 1. where relevant, medication within the last four weeks before sampling (when sampling on farm);

• 1. substance or substance groups for examination;

• 1. particular remarks.

Paper or electronic copies of the report are to be provided depending on the sampling procedure. The sampling report and its copies shall be completed in a way that ensures their authenticity and legal validity, which may require that these documents are signed by the inspector. In case of on-farm sampling, the farmer or his deputy may be invited to sign the original sampling report.

The original of the sampling report remains at the competent authority, which has to guarantee that unauthorised persons cannot access this original report.

If necessary, the farmer or the owner of the establishment may be informed of the sampling undertaken.

6. Sampling report for the laboratory

The sampling report for the laboratory established by the competent authorities shall be in accordance with the requirements set in Chapter 7 of ISO/IEC 17025:2017 ([^E0043]) and shall contain at least the following information:

• 1. address of the competent authorities or designated bodies;

• 1. name of inspector or identification code;

• 1. official code number of the sample;

• 1. sampling date;

• 1. animal species;

• 1. sample matrix;

• 1. substances or substance groups for examination;

• 1. particular remarks.

The sampling report for the laboratory shall accompany the sample when sent to the laboratory.

7. Transport and storage

Residue control programmes shall specify the suitable storage and transport conditions for each analyte/matrix combination to ensure analyte stability and sample integrity. The transport time shall be as short as possible and the temperature during transport shall be adequate to ensure analyte stability.

Specific attention shall be paid to transport boxes, temperature and delivery times to the responsible laboratory.

In case of any non-compliance with the requirements of the control programme, the laboratory shall inform the competent authority without delay.

[^E0001]: Commission Implementing Regulation (EU) 2022/1646 of 23 September 2022 on uniform practical arrangements for the performance of official controls as regards the use of pharmacologically active substances authorised as veterinary medicinal products or as feed additives and of prohibited or unauthorised pharmacologically active substances and residues thereof, on specific content of multi-annual national control plans and specific arrangements for their preparation (OJ L 248, 26.9.2022, p. 32, ELI: http://data.europa.eu/eli/reg_impl/2022/1646/oj).

[^E0002]: Commission Delegated Regulation (EU) 2019/2090 of 19 June 2019 supplementing Regulation (EU) 2017/625 of the European Parliament and Council regarding cases of suspected or established non-compliance with Union rules applicable to the use or residues of pharmacologically active substances authorised in veterinary medicinal products or as feed additives or with Union rules applicable to the use or residues of prohibited or unauthorised pharmacologically active substances (OJ L 317, 9.12.2019, p. 28).

[^E0003]: Commission Regulation (EU) 2019/1871 of 7 November 2019 on reference points for action for non-allowed pharmacologically active substances present in food of animal origin and repealing Decision 2005/34/EC (OJ L 289, 8.11.2019, p. 41).

[^E0004]: Regulation (EC) No 470/2009 of the European Parliament and of the Council of 6 May 2009 laying down Community procedures for the establishment of residue limits of pharmacologically active substances in foodstuffs of animal origin, repealing Council Regulation (EEC) No 2377/90 and amending Directive 2001/82/EC of the European Parliament and of the Council and Regulation (EC) No 726/2004 of the European Parliament and of the Council (OJ L 152, 16.6.2009, p. 11).

[^E0005]: Council Regulation (EEC) No 315/93 of 8 February 1993 laying down Community procedures for contaminants in food (OJ L 37, 13.2.1993, p. 1).

[^E0006]: ISO 3534-1: 2006 Statistics – Vocabulary and symbols – Part 1: General statistical terms and terms used in probability (Chapter 1).

[^E0007]: Directive 2001/82/EC of the European Parliament and of the Council of 6 November 2001 on the Community code relating to veterinary medicinal products (OJ L 311, 28.11.2001, p. 1).

[^E0008]: JCGM 200:2008, International vocabulary of metrology – Basic and general concepts and associated terms (VIM), Third Edition 2008: https://www.iso.org/sites/JCGM/VIM-JCGM200.htm (Chapter 5 Measurement standards (Etalons)).

[^E0009]: Commission Regulation (EC) No 124/2009 of 10 February 2009 setting maximum levels for the presence of coccidiostats or histomonostats in food resulting from the unavoidable carry-over of these substances in non-target feed (OJ L 40, 11.2.2009, p. 7).

[^E0010]: Commission Regulation (EU) No 37/2010 of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin (OJ L 15, 20.1.2010, p. 1).

[^E0011]: Codex Alimentarius Commission, Food and Agriculture Organization of the United Nations/World Health Organization, Guidelines on analytical terminology (CAC/GL 72-2009).

[^E0012]: ISO 5725-1:2023 Accuracy (trueness and precision) of measurement methods and results – Part 1: General principles and definitions (Chapter 3).

[^E0013]: ISO 80000-1:2022 Quantities and units – Part 1: General (Introduction).

[^E0014]: Council Directive 80/181/EEC of 20 December 1979 on the approximation of the laws of the Member States relating to units of measurement and on the repeal of Directive 71/354/EEC (OJ L 39, 15.2.1980, p. 40, ELI: http://data.europa.eu/eli/dir/1980/181/oj).

[^E0015]: ISO/IEC 17025:2017 General requirements for the competence of testing and calibration laboratories (Chapter 3).

[^E0016]: ISO 78-2: 1999 Chemistry – Layouts for standards – Part 2: Methods of chemical analysis (Annexes).

[^E0017]: Co-chromatography is a procedure in which the sample extract prior to the chromatographic step(s) is divided into two parts. Part one is chromatographed as such. Part two is mixed with the standard analyte that is to be measured. Then this mixture is also chromatographed. The amount of added standard analyte has to be similar to the estimated amount of the analyte in the extract. Co-chromatography is used to improve the identification of an analyte when chromatographic methods are used, especially when no suitable internal standard can be used.

[^E0023]: ISO 5725-4:2020 Accuracy (trueness and precision) of measurement methods and results – Part 4: Basic methods for the determination of the trueness of a standard measurement method (Clause 3).

[^E0024]: Where, for a non-allowed pharmacologically active substance validation of a concentration of 0,5 times the RPA is not reasonably achievable, the concentration of 0,5 times the RPA can be replaced by the lowest concentration between 0,5 times and 1,0 times the RPA, which is reasonably achievable or by the LCL in case that it is lower than 0,5 times the RPA.

[^E0025]: Where, for a specific pharmacologically active substance validation of a concentration of 0,1 times the MRL is not reasonably achievable, the concentration of 0,1 times the MRL can be replaced by the lowest concentration between 0,1 times and 0,5 times the MRL, which is reasonably achievable.

[^E0026]: Where, for a non-allowed pharmacologically active substance validation of a concentration of 0,5 times the RPA is not reasonably achievable, the concentration of 0,5 times the RPA can be replaced by the lowest concentration between 0,5 times and 1,0 times the RPA, which is reasonably achievable or by the LCL in case that it is lower than 0,5 times the RPA.

[^E0027]: ISO 5725-2:2019 Accuracy (trueness and precision) of measurement methods and results – Part 2: Basic method for the determination of repeatability and reproducibility of a standard measurement method (Clause 3).

[^E0028]: Where, for a non-allowed pharmacologically active substance validation of a concentration of 0,5 times the RPA is not reasonably achievable, the concentration of 0,5 times the RPA can be replaced by the lowest concentration between 0,5 times and 1,0 times the RPA, which is reasonably achievable or by the LCL in case that it is lower than 0,5 times the RPA.

[^E0029]: ISO 11843-1:1997 Capability of detection – Part 1: Terms and definitions.

[^E0030]: Codex Alimentarius Commission, Food and Agriculture Organization of the United Nations, World Health Organization, Guidelines on estimation of uncertainty of results (CAC/GL 59-2006).

[^E0031]: The changes in experimental conditions referred to therein can consist of the sample materials, analytes, storage conditions, environmental and/or sample preparation conditions. For all experimental conditions, which could in practice be subject to fluctuation (e.g. stability of reagents, composition of the sample, pH, temperature) any variations which could affect the analytical result shall be indicated.

[^E0032]: Jülicher, B., Gowik, P. and Uhlig, S. (1998) Assessment of detection methods in trace analysis by means of a statistically based in-house validation concept. Analyst, 123, 173.

[^E0033]: ISO/TS 23471:2022 Experimental designs for evaluation of uncertainty – Use of factorial designs for determining uncertainty functions.

[^E0034]: IUPAC (1995), Protocol for the design, conduct and interpretation of method-performance studies, Pure & Applied Chem, 67, 331.

[^E0035]: Gowik, P., Jülicher, B. and Uhlig, S. (1998) Multi-residue method for non-steroidal anti-inflammatory drugs in plasma using high performance liquid chromatography-photodiode-array detection. Method description and comprehensive in-house validation. J. Chromatogr., 716, 221.

[^E0036]: Lamberty, A., Schimmel, H. and Pauwels, J. (1998) The study of the stability of reference materials by isochronous measurements. Fres. J. Anal. Chem. 360, 359.

[^E0037]: ISO 11843-1:1997 Capability of detection – Part 1: Terms and definitions.

[^E0038]: Commission Implementing Regulation (EU) 2018/470 of 21 March 2018 on detailed rules on the maximum residue limit to be considered for control purposes for foodstuffs derived from animals which have been treated in the EU under Article 11 of Directive 2001/82/EC (OJ L 79, 22.3.2018, p. 16).

[^E0039]: The amount of the standard analyte added, can be, for example, between two and five times the estimated amount of the analyte in the sample. This procedure is designed to determine the content of an analyte in a sample, taking account of the recovery of the analytical procedure.

[^E0040]: ISO/IEC 17025: 2017 General requirements for the competence of testing and calibration laboratories (Chapter 7.7).

[^E0041]: ISO/IEC 17043:2010 Conformity assessment – General requirements for proficiency testing.

[^E0042]: ISO/IEC 17043:2023 Conformity assessment – General requirements for proficiency testing.

[^E0043]: ISO/IEC 17025: 2017 General requirements for the competence of testing and calibration laboratories (Chapter 7.7).