Commission Implementing Regulation (EU) No 503/2013 of 3 April 2013 on applications for authorisation of genetically modified food and feed in accordance with Regulation (EC) No 1829/2003 of the European Parliament and of the Council and amending Commission Regulations (EC) No 641/2004 and (EC) No 1981/2006 Text with EEA relevance

• (a) the composition of the genetically modified food and feed with regard to the levels of nutrients and anti-nutrients (see compositional studies as described in Section 1.3);

• (b) the bioavailability and biological efficacy of nutrients in the food and feed taking into account the potential influences of transport, storage and expected treatment of the food and feed;

• (c) the anticipated dietary intake of the food and feed (see Section 2) and resulting nutritional impact.

When the comparative analysis has identified compositional characteristics of the genetically modified food and feed that are different from its conventional counterpart and/or not equivalent to the characteristics of the reference varieties, their nutritional relevance shall be assessed on the basis of current scientific knowledge. If that assessment does conclude to the nutritional equivalence between the genetically modified food and feed and its conventional counterpart, no further studies shall be carried out. By contrast if, on the basis of the assessment of the information obtained from the comparative analysis, it is not possible to conclude on the nutritional equivalence, further nutritional studies shall be carried out. Comparative growth studies shall be conducted with young rapidly growing animal species (such as broiler chicks as animal model for non-ruminants; lambs for ruminants; or other rapidly growing species).

1.6.3.   Nutritional studies of genetically modified food

The applicant shall determine the necessity and design of nutritional studies on the basis of the introduced trait(s), the outcome of the comparative analysis, and of the 90-day feeding study, where available. Supplementary information regarding the nutritional value may be obtained from comparative growth performance studies conducted with other animal species, such as broiler chickens, addressing the nutritional assessment of genetically modified feed. When nutritional studies are conducted, the control diet(s) shall include the conventional counterpart and where appropriate additional comparator(s). In the case of herbicide tolerant genetically modified plants, the tested material should come from the genetically modified plant exposed to the intended herbicide.

Genetically modified foods modified to provide additional health benefits to the consumer as compared to conventional foods, may benefit specific populations or subpopulations while others may be at risk from the same food. In cases where an altered bioavailability needs to be established and may raise concern for subpopulation(s), the level of the nutrient in the food shall be determined, taking into account all the different forms of the compound. The methods to test for bioavailability shall be selected on a case-by-case basis depending on the nutrient or other constituent, the food containing these constituents, as well as the health, nutritional status and dietary practices of the specific population(s) anticipated to consume the food.

1.6.4.   Nutritional studies of genetically modified feed

The applicant shall determine the necessity and design of further nutritional studies on the basis of the introduced trait(s), the outcome of the comparative analysis, and the 90-day feeding study, where available. Supplementary information regarding the nutritional value may be obtained from comparative growth performance studies conducted with other animal species, such as broiler chickens, addressing the nutritional assessment of genetically modified feed. When nutritional studies are conducted, the control diet(s) shall include the conventional counterpart and where appropriate additional comparator(s).

In the case of genetically modified feed with improved nutritional characteristics, feeding studies with target animal of food producing species shall be conducted to assess the impact on the feed. In the case of genetically modified plants modified for improved content and bioavailability of nutrients, studies with target food producing animal species shall be conducted to determine the bioavailability of individual nutrients in the genetically modified plant compared to its conventional counterpart. In the case of genetically modified plants specifically modified with traits to enhance animal performance through increased nutrient density (such as increased oil content) or an enhanced level of a specific nutrient (such as an essential amino acid or a vitamin), an appropriate control diet using its conventional counterpart shall be formulated by supplementing it with the specific nutrient to the extent of the change effected in the genetically modified plant. Regarding co-products (such as oilseeds meals), from which the ingredient targeted by the genetic modification has been extracted, these may be compared with co-products produced from the conventional counterpart.

Target animal feeding studies shall span either the growing and/or finishing period to slaughter for chickens, pigs, and cattle for fattening or a major part of a lactation cycle for dairy cows, or laying cycle for laying hens or quails. For feedstuffs intended only for aquaculture, growth studies with aquatic species such as carp, catfish, salmonidae or typical herbivores shall be chosen.

When appropriate, tests with various experimental designs shall be provided to demonstrate that the nutritionally improved genetically modified plant fulfils the expected nutritional value. The exact experimental design and statistical approaches of feeding experiments in food producing animals to test the nutritional value of genetically modified feed, modified for enhanced nutritional characteristics, shall depend on the targeted animal species, type of plant trait(s) studied and the size of the expected effect. The experimental diets shall be formulated in such a way that the key measured endpoints are responsive to a difference in the quantity and/or availability of the nutrient in question. Endpoint measurements shall vary with the target species used in the study, but shall include feed intake, body weight, animal performance and bioavailability of nutrients.

More detailed guidance for the application of the requirements of this Section is available in the Report of the EFSA GMO Panel Working Group on Animal Feeding Trials [^ntr10-L_2013157EN.01001901-E0010].

1.6.5.   Conclusion of the nutritional assessment

The conclusion of the nutritional assessment of genetically modified food and feed shall indicate whether the genetically modified food and feed is nutritionally equivalent to its conventional counterpart, taking natural variations into account.

The applicant shall evaluate the result of the nutritional assessment in the light of anticipated intake of the genetically modified food and feed (see Section 2).

1.7.   Standardised guidelines for toxicity tests

The applicant shall use for toxicity testing internationally agreed guidelines and test methods described by Commission Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) [^ntr11-L_2013157EN.01001901-E0011] (see Tables 1 and 2). A non-exhaustive list of validated test methods which, where necessary, shall be used in a possibly adapted form for GMO toxicological testing is provided in Tables 1 and 2.

The performance of test methods depends on the type of genetically modified food and feed, type of the genetic modification and resulting intended and unintended alterations, intended use and exposure/intake, and the available knowledge. Some of the tests were developed for the assessment of risks at the workplace (see Sections 1.4 and 1.5).

Table 1

Non-exhaustive list of validated test methods for chemicals as provided by Regulation (EC) No 440/2008 and which may be used in a possibly adapted form for GMO toxicological testing

Table 2

Genotoxicity tests as provided by Regulation (EC) No 440/2008

2. EXPOSURE ASSESSMENT — ANTICIPATED INTAKE/EXTENT OF USE

An estimate of the expected intake shall be an essential element in the risk assessment of genetically modified food and feed and shall also be required for the nutritional evaluation. Information shall be provided by the applicant on the intended function, the dietary role, and the expected level of use of the genetically modified food and feed in the EU. In addition, the expected range of concentrations of newly produced proteins or existing plant proteins deliberately modified in the genetically modified food(s) and feed(s) to be placed on the market shall be provided.

On the basis of representative consumption data for products obtained from the respective conventional plants, the applicant shall estimate the anticipated average and maximum intake of the genetically modified food and feed. Probabilistic methods may be used to determine ranges of plausible values rather than single values or point estimates. The applicant shall identify and consider particular groups of the EU population with an expected higher exposure and shall consider this higher exposure within the risk assessment. Any assumptions made in the exposure assessment shall be described. Recent developments in methodologies and appropriate consumption data shall be used. Data on import and production quantities may provide additional information for the intake assessment.

The applicant shall determine by appropriate methods the concentrations of the newly expressed proteins, other new constituents and endogenous food and feed constituents, of which the levels have been altered as a result of the genetic modification (for example, due to changes in metabolic pathways) in those parts of the genetically modified plant intended for food or feed use. Expected intake of these constituents shall be estimated taking into account the influences of processing, storage and expected treatment of the food and feed in question, for example, potential accumulation or reduction. In cases where the genetic modification has resulted in an altered level of a natural constituent, or if a new constituent occurs naturally in other food and feed products, the anticipated change in total intake of this constituent shall be assessed considering realistic as well as worst case intake scenarios.

The applicant shall provide information on known or anticipated human/animal intake of analogous genetically modified food and feed and on other routes of exposure to the respective new and natural constituents, including amount, frequency and other factors influencing exposure.

3. RISK CHARACTERISATION

3.1.   Introduction

The applicant shall base its risk characterisation of genetically modified plants and foods and feed on data from hazard identification, hazard characterisation, and on exposure/intake data. The applicant shall ensure that the risk characterisation is comprehensive by considering all the available evidence from several analysis including molecular analysis, phenotypic, agronomical and compositional analysis, toxicity and allergenicity testing. The applicant shall consider indications resulting from the risk characterisation that may require specific activities for post-market monitoring of genetically modified food and feed.

In performing its risk characterisation, the applicant shall demonstrate that the hazard identification and hazard characterisation are complete. The applicant shall discuss the quality of existing data and information. The discussion shall clearly indicate how this body of information has been taken into account in the determination of the final risk characterisation.

The applicant shall provide estimations of the uncertainties associated to each test as well as to the different stages of the risk assessment. The applicant shall quantify them to the possible extent. A distinction shall be made between uncertainties that reflect natural variations in biological parameters (including variations in susceptibility in populations), and variation amongst different species’ responses.

Depending on the issue to be addressed and the available data, the applicant shall perform a qualitative and, where possible, quantitative risk characterisation. The conditions for the estimated risk, and associated uncertainties, shall be as precise as possible.

3.2.   Issues to be considered for risk characterisation

When appropriate and depending on the type of genetic modification, the applicant shall carry out a risk assessment of genetically modified plants in an integrative manner in accordance with Section 3.1. This risk assessment shall be performed on a case-by-case basis depending on the modified plant and the type of genetic modification, the cultivation practices of the genetically modified plant and uses of the genetically modified food and feed. The applicant shall take into account the different issues considered in hazard identification and hazard characterisation and exposure steps. The outcomes of these issues shall be considered by the applicant together in the risk characterisation step. The list of issues provided in this Section shall not be exhaustive.

3.2.1.   Molecular characterisation

Evaluation of the characteristics and previous use of the donor and the recipient plant shall be a key element to identify the need for specific analyses, such as the occurrence of specific toxins, or allergens in the unmodified recipient plant which may be unintentionally increased as result of the genetic modification.

Transformation protocols, molecular characterisation strategies and the specificity and sensitivity of the methods used shall be discussed by the applicant in relation to the intentional and possibly unintentional insertion and expression of gene sequences.

Where sequence analysis has identified a potential hazard, the applicant shall demonstrate how approaches like bioinformatic analysis, compositional/agronomical analysis and possibly animal feeding trials with the whole genetically modified food and feed contribute to the safety assessment. The value of the results obtained shall be evaluated in the light of the available knowledge on the structure and function of genomic databases of the crop species in question or related species.

In cases of GM plants containing stacked transformation events, the additional risks possibly arising from the combined effects of the stacked genes shall be evaluated.

3.2.2.   Comparative analysis

The first goal of the comparative analysis is to identify possible differences between the GM plant and its conventional counterpart and, where appropriate, additional comparators. The second goal of the comparative analysis is to identify possible lack of equivalences between the GM plant and its reference varieties. These differences and/or lack of equivalences should be assessed with respect to their possible impact on food and feed safety and nutritional properties, taking into account natural variation. The estimated risk and associated uncertainties should be as precise as possible and taken into account.

The applicant shall demonstrate that the comparative analysis between the genetically modified plant and its conventional counterpart with respect to agronomic, morphological and compositional characteristics has been carried out in accordance with the requirements of this Regulation. The selection of the conventional counterpart and, where appropriate, additional comparators shall be justified.

3.2.3.   Food and feed safety in relation to intake

The applicant shall evaluate the data generated to estimate possible short-term and long-term risks to human or animal health associated with the consumption of genetically modified food or feed with respect to the expression of new proteins/metabolites, as well as significantly altered levels of original plant proteins/metabolites in genetically modified foods/feed. This evaluation shall include a thorough analysis of the relevance and limitations of each test as well as of the whole information.

The applicant shall consider the range of levels observed for the compounds known to occur in the conventional counterpart and in reference varieties. This variability may be caused by differences that are genotype-dependent, environmentally dependent, or caused by genotype x environment interactions. In addition, the range of levels observed in a broad spectrum of food and feed representative for the human and animal diet may be taken into account given that it reflects the levels of the specific compound to which consumers may be exposed.

If single constituents and/or whole genetically modified food and feed are found to induce adverse effects in specific studies, information on dose response relationships, threshold levels, delayed onset of adverse effects, risks for certain groups in the population, and use of uncertainty factors in extrapolation of animal data to humans shall be submitted.

The applicant shall consider data on the characteristics of the new compounds present in the genetically modified plant including potential biological effects in humans and animals. If the compounds have known adverse health effects and maximum levels for the presence of these compounds in the plant or its products were laid down in specific legislation, these maximum levels shall be taken into account. Otherwise, reference values for acceptable or tolerable levels of intake, such as the acceptable daily intake (ADI) or tolerable upper intake level (UL), shall be considered in relation to the anticipated intake. In cases where the compound has been safely consumed in food, the intake levels of consumers from a conventional diet shall be considered as safe.

The applicant shall evaluate the information on the effects of processing on the new compounds. Potential accumulation/depletion in food and feed products entering the human or animal diet shall be considered. The applicant shall also evaluate the relevance of differences resulting from chemical reactions known to occur under processing conditions.

In cases where more complex genetic modifications are produced, for example, via transfer of multiple genes in a single construct, retransformation of pre-existing genetically modified lines, and stacking of transformation events through conventional breeding of genetically modified parents, the applicant shall discuss strategies for the assessment of any risk(s) associated with possible interactions between the newly expressed proteins, new metabolites and original plant constituents. The assessment shall take into account all available information including the mode of action of the newly expressed proteins, the molecular and compositional/agronomical characteristics of the genetically modified plant, and the outcome of animal toxicity studies and feeding trials.

The applicant shall evaluate data obtained to assess the allergenic potential of newly expressed proteins in genetically modified plants with respect to introduction of new allergenic proteins into the food and feed plants, a possible provocation of allergic reactions of susceptible individuals, as well as information to demonstrate that the genetic modification does not cause unintended changes in the characteristics and/or levels of expression of endogenous allergenic proteins in the genetically modified food. In particular, the choice of the test models shall be justified with respect to specificity, predictability and validation status.

With respect to intake estimations of genetically modified foods, the applicant shall evaluate the applied methodologies with respect to uncertainties associated with the prediction of long-term intake. Specific attention shall be paid to those genetically modified plants which are aimed at modifying nutritional characteristics of the food and feed. For those genetically modified products, the requirement for post-market monitoring shall be discussed as a mechanism for determining actual changes to overall dietary intake patterns of the genetically modified food, to what extent this has occurred and whether or not the product induces known (side) effects or unexpected side effects. If the performance of post-market monitoring is deemed necessary, the reliability, sensitivity and specificity of the proposed methods shall be provided.

3.3.   The result of risk characterisation

In accordance with the requirements of Articles 4 and 16 of Regulation (EC) No 1829/2003, the applicant shall ensure that the final risk characterisation clearly demonstrates that:

• (a) the genetically modified food and feed has no adverse effects on human and animal health;

• (b) the genetically modified food does not differ from the food which it is intended to replace to such an extent that its normal consumption would be nutritionally disadvantageous for the consumer;

• (c) the genetically modified food does not mislead the consumer;

• (d) the genetically modified feed does not harm or mislead the consumer by impairing the distinctive features of the animal products;

• (e) the genetically modified feed does not differ from the feed which it is intended to replace to such an extent that its normal consumption would be nutritionally disadvantageous for animals or humans.

The applicant shall clearly indicate what assumptions have been made during the risk assessment in order to predict the probability of occurrence and severity of adverse effect(s) in a given population, and the nature and magnitude of uncertainties associated with establishing these risks.

The applicant shall also include detailed information justifying the inclusion or not of a proposal for labelling in the application, in accordance with Article 13(2)(a), Article 13(3), Article 25(2)(c) and Article 25(3) of Regulation (EC) No 1829/2003.

[^ntr1-L_2013157EN.01001901-E0001]: OJ L 43, 14.2.1997, p. 1.

[^ntr2-L_2013157EN.01001901-E0002]: EFSA Journal 2011; 9(5):2149.

[^ntr3-L_2013157EN.01001901-E0003]: EFSA Journal 2010; 8(1):1250.

[^ntr4-L_2013157EN.01001901-E0004]: EFSA Journal 2010; 8(1):1250.

[^ntr7-L_2013157EN.01001901-E0007]: OJ L 133, 22.5.2008, p. 1.

[^ntr8-L_2013157EN.01001901-E0008]: EFSA Journal 2011; 9(12):2438.

[^ntr9-L_2013157EN.01001901-E0009]: EFSA Journal 2010; 8(7):1700.

[^ntr10-L_2013157EN.01001901-E0010]: EFSA, 2008 Report of the EFSA GMO Panel Working Group on Animal Feeding Trials, 2008. Safety and nutritional assessment of GM plants and derived food and feed. The role of animal feeding trials. Food and Chemical Toxicology 46 (2008) S2-S70.

[^ntr11-L_2013157EN.01001901-E0011]: OJ L 142, 31.5.2008, p. 1.

ANNEX III

VALIDATION OF METHODS FOR THE DETECTION, IDENTIFICATION AND QUANTIFICATION OF THE TRANSFORMATION EVENT, AND REQUIREMENTS FOR CONTROL SAMPLES AND THE CERTIFIED REFERENCE MATERIAL

1. INTRODUCTION

2. DEFINITIONS

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

• (a) ‘certified reference material’ means reference material as referred to in Articles 5(3)(j) and 17(3)(j) of Regulation (EC) No 1829/2003 and corresponds to any material or substance, one or more of whose property values are certified for calibration or quality control of methods. It is accompanied by a certificate that provides value of the specified property, its associated uncertainty and a statement of metrological traceability;

• (b) ‘method performance requirements’ means the minimum performance criteria that the method shall demonstrate upon completion of the validation study carried out by the EURL, according to internationally accepted technical provisions.

3. METHOD VALIDATION

3.1.   Information about the method

3.2.   Information about the method testing carried out by the applicant

3.3.   Samples of the food and feed and their control samples

For the purpose of implementing Articles 5(3)(j) and 17(3)(j) of Regulation (EC) No 1829/2003, the applicant shall, together with the information required under Sections 1, 2 and 3 of this Annex, also provide samples of the food and feed and their control samples of a type and amount to be specified by the EURL for the specific application for authorisation.

The information accompanying the control samples shall include information on the breeding of the plant which has been used for the production of the control samples and on the zygosity of the insert(s).

The applicant may use the same raw material for the production of certified reference material and for the production of control samples.

4. CERTIFIED REFERENCE MATERIAL

The certified reference material shall be produced under ISO Guide 34 (General requirements for the competence of reference material producers) by a producer accredited to ISO Guide 34.

The applicant shall provide information as regards the place where the certified reference material can be accessed. This shall be accompanied by adequate information demonstrating that the availability of the certified reference material will be maintained throughout the period of validity of the authorisation. For verification and value assignment, a method that has been properly validated (see ISO/IEC 17025: General requirements for the competence of testing and calibration laboratories) shall be used.

Uncertainties shall be estimated according to the ISO Guide to the Expression of Uncertainty in Measurement (GUM).

The main characteristics of those internationally accepted technical provisions are the following:

• 1. Genetically modified reference material containers:

• (a) genetically modified reference material container (such as bottles, vials, ampoules) shall be tight and contain not less than the stated amount of material;

• (b) the commutability of the genetically modified reference material must be assured;

• (c) packaging shall be appropriate to the purpose;

• (d) labelling shall be of good aspect and quality.

• 1. Homogeneity testing:

• (a) samples shall have appropriate homogeneity;

• (b) between-bottle homogeneity shall be examined;

• (c) any possible between-bottle heterogeneity shall be accounted for in the overall estimated reference material uncertainty. This requirement shall apply even when no statistically significant between-bottle variation is present. In this case, the method variation or the actual calculated between-bottle variation, whichever is larger, shall be included in the overall uncertainty.

• 1. Stability testing:

• (a) samples shall have appropriate stability;

• (b) stability shall be positively demonstrated by appropriate statistical extrapolation for the genetically modified reference material shelf-life to be within the stated uncertainty; the uncertainty related to this demonstration is part of the estimated reference material uncertainty. Assigned values are valid only for a limited time and shall be subject to a stability monitoring.

• 1. Batch characterisation:

• 1. The methods used for verification and for certification shall:

• (a) be applied under metrologically valid conditions;

• (b) have been properly technically validated before use;

• (c) have precision and trueness compatible with the target uncertainty.

• 1. Each set of measurements shall:

• (a) be traceable to the stated references;

• (b) be accompanied by an uncertainty statement whenever possible.

• 1. Participating laboratories shall:

• (a) have the required competence for the execution of the task;

• (b) be able to achieve traceability to the required stated references;

• (c) be able to estimate their measurement uncertainty;

• (d) have in place a sufficient and appropriate quality assurance system.

• 1. Final storage:

• 1. To avoid degradation after sample production, all samples shall be stored under conditions designated for the final storage of the genetically modified certified reference material before measurements are started.

• 1. Otherwise, they shall be transported from door to door keeping them at all times under such storage conditions for which it has been demonstrated that there is no influence on the assigned values.

• 1. Establishment of a certificate for certified reference material:

A certificate complemented by a certification report shall be established, containing all information relevant to and needed by the user.

The certificate and report shall be made available when the genetically modified certified reference material is distributed.

The information accompanying the certified reference material shall include information on the breeding of the plant which has been used for the production of the certified reference material and on the zygosity of the insert(s).

The certified value of the GMO content shall be given in mass fraction and, where available, in copy number per haploid genome equivalent.

Certified values (such as quantity of genetically modified material expressed in mass fraction) shall be traceable to stated references and be accompanied by an expanded uncertainty statement valid for the entire shelf-life of the genetically modified certified reference material.