COMMISSION REGULATION (EU) No 544/2011

of 10 June 2011

implementing Regulation (EC) No 1107/2009 of the European Parliament and of the Council as regards the data requirements for active substances

(Text with EEA relevance)

THE EUROPEAN COMMISSION,

Having regard to the Treaty on the Functioning of the European Union,

Having regard to Regulation (EC) No 1107/2009 of the European Parliament and of the Council of 21 October 2009 concerning the placing of plant protection products on the market and repealing Council Directives 79/117/EEC and 91/414/EEC (1), and in particular the first sentence of Article 8(4) thereof,

After consulting the Standing Committee on the Food Chain and Animal Health,

Whereas:

In accordance with Regulation (EC) No 1107/2009, the dossier to be submitted for the approval of an active substance or for the authorisation of a plant protection product is to fulfil the same requirements in respect of the data requirements for the plant protection product as under the previously applicable rules which are set out in Annexes II and III to Council Directive 91/414/EEC of 15 July 1991 concerning the placing of plant protection products on the market (2) .

It is therefore necessary for the implementation of Regulation (EC) No 1107/2009 to adopt a regulation containing those data requirements for active substances. Such a regulation is not to include any substantial modification,

HAS ADOPTED THIS REGULATION:

Article 1

The data requirements for the approval of an active substance provided for in Article 8(1)(b) of Regulation (EC) No 1107/2009 shall be as set out in the Annex to this Regulation.

Article 2

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 14 June 2011.

This Regulation shall be binding in its entirety and directly applicable in all Member States.

(1) OJ L 309, 24.11.2009, p. 1.

(2) OJ L 230, 19.8.1991, p. 1.

ANNEX

DATA REQUIREMENTS FOR ACTIVE SUBSTANCES, AS PROVIDED FOR IN ARTICLE 8(1)(b) OF REGULATION (EC) No 1107/2009

INTRODUCTION

1. The information required shall:

1.1. include a technical dossier supplying the information necessary for evaluating the foreseeable risks, whether immediate or delayed, which the active substance may entail for humans, animals and the environment and containing at least the information and results of the studies referred to below;

1.2. where relevant, be generated using test guidelines, in accordance with the latest adopted version, referred to or described in this Annex; in the case of studies initiated before the entry into force of the modification of this Annex, the information shall be generated using suitable internationally or nationally validated test guidelines or, in the absence thereof, test guidelines accepted by the competent authority;

1.3. in the event of a test guideline being inappropriate or not described, or where another one than those referred to in this Annex has been used, include a justification, which is acceptable to the competent authority for the guidelines used. In particular, when reference is made in this Annex to a method laid down in Commission Regulation (EC) No 440/2008 (1) which consists in the transposition of a method developed by an international organisation (e.g. OECD), Member States may accept that the required information is generated in accordance with the latest version of that method if at the initiation of the studies the method under Regulation (EC) No 440/2008 has not yet been updated;

1.4. include, when required by the competent authority, a full description of test guidelines used, except if they are referred to or described in this Annex, and a full description of any deviations from them including a justification, which is acceptable to the competent authority, for these deviations;

1.5. include a full and unbiased report of the studies conducted as well as full description of them or a justification, which is acceptable to the competent authority where: — particular data and information which would not be necessary owing to the nature of the product or its proposed uses, are not provided, or — it is not scientifically necessary, or technically possible to supply information and data;

1.6. where relevant, have been generated in accordance with the requirements of Council Directive 86/609/EEC (2).

2. Test and analyses

2.1. Tests and analyses must be conducted in accordance with the principles laid down in Directive 2004/10/EC of the European Parliament and of the Council (3) where testing is done to obtain data on the properties and/or safety with respect to human or animal health or the environment.

2.2. By way of derogation from point 2.1, Member States may provide that tests and analyses, performed on their territory in order to obtain data on the properties and/or safety of the active substances with respect to honey-bees and beneficial arthropods other than bees shall be conducted by official or officially-recognised testing facilities or organisations which satisfy at least the requirements as set out under points 2.2 and 2.3 of the introduction to the Annex to Commission Regulation (EU) No 545/2011 (4). This derogation applies to trials which actually started on or before 31 December 1999.

2.3. By way of derogation from point 2.1, Member States may provide that supervised residue trials performed on their territory in accordance with Section 6 ‘Residues in or on treated products, food and feed’ on plant protection products containing active substances already on the market 2 years after notification of the Directive 91/414/EEC shall be conducted by official or officially-recognised testing facilities or organisations which satisfy at least the requirements under points 2.2 and 2.3 of the introduction to the Annex to Regulation (EU) No 545/2011. This derogation applies for supervised residue trials which actually started on or before 31 December 1997.

2.4. By way of derogation from point 2.1, for active substances consisting of micro-organisms or viruses, tests and analyses done to obtain data on the properties and/or safety with respect to other aspects than human health, may have been conducted by official or officially recognised testing facilities or organisations which satisfy at least the requirements under points 2.2 and 2.3 of the introduction to the Annex to Regulation (EU) No 545/2011.

PART A

CHEMICAL SUBSTANCES

1. Identity of the active substance

The information provided must be sufficient to identify with precision each active substance, to define it in terms of its specification and to characterise it as to its nature. The information and data referred to, unless otherwise specified, are required for all active substances.

1.1. Applicant (name, address, etc.)

The name and address of the applicant must be provided as must the name, position, telephone and telefax number of the appropriate person to contact.

Where, in addition, the applicant has an office, agent or representative in the Member State to which the application for approval is submitted, and if different, in the rapporteur Member State appointed by the Commission, the name and address of the local office, agent or representative must be provided, as must the name, position, telephone and telefax number of the appropriate person to contact.

1.2. Manufacturer (name, address, including location of plant)

The name and address of the manufacturer or manufacturers of the active substance must be provided as must the name and address of each manufacturing plant in which the active substance is manufactured. A contact point (preferably a central contact point, to include name, telephone and telefax number) must be provided, with a view to providing updating information and responding to queries arising, regarding manufacturing technology, processes and the quality of product (including where relevant, individual batches). Where following approval of the active substances, there are changes in the location or number of manufacturers, the information required must again be notified to the Commission and the Member States.

1.3. Common name proposed or ISO-accepted, and synonyms

The ISO common name, or proposed ISO common name and where relevant, other proposed or accepted common names (synonyms), including the name (title) of the nomenclature authority concerned, must be provided.

1.4. Chemical name (IUPAC and CA nomenclature)

The chemical name as given in Annex VI to Regulation (EC) No 1272/2008 of the European Parliament and of the Council (5), or, if not included in that Regulation, in accordance with both IUPAC and CA nomenclature, must be provided.

1.5. Manufacturer’s development code number(s)

Code numbers used to identify the active substance, and where available, formulations containing the active substance, during development work, must be reported. For each code number reported, the material to which it relates, the period for which it was used, and the Member States or other countries in which it was used and is being used, must be stated.

1.6. CAS, EC and CIPAC numbers (if available)

Chemical Abstracts, EC (Einecs or ELINCS), and CIPAC numbers, where they exist, must be reported.

1.7. Molecular and structural formula, molecular mass

The molecular formula, molecular mass and structural formula of the active substance, and where relevant, the structural formula of each stereo and optical isomer present in the active substance, must be provided.

1.8. Method of manufacture (synthesis pathway) of the active substance

The method of manufacture, in terms of the identity of the starting materials, the chemical pathways involved, and the identity of by-products and impurities present in the final product, must be provided, for each manufacturing plant. Generally process engineering information is not required.

Where the information provided relates to a pilot plant production system, the information required must again be provided once industrial scale production methods and procedures have stabilised.

1.9. Specification of purity of the active substance in g/kg

The minimum content in g/kg of pure active substance (excluding inactive isomers) in the manufactured material used for production of formulated products, must be reported.

Where the information provided relates to a pilot plant production system, the information required must again be provided to the Commission and the Member States once industrial scale production methods and procedures have stabilised, if production changes result in a changed specification of purity.

1.10. Identity of isomers, impurities and additives (e.g. stabilisers), together with the structural formula and the content expressed as g/kg

The maximum content in g/kg of inactive isomers as well as the ratio of the content of isomers/diastereo-isomers, where relevant, must be provided. In addition, the maximum content in g/kg of each further component other than additives, including by-products, and impurities, must be provided. In the case of additives the content in g/kg must be provided.

For each component, present in quantities of 1 g/kg or more, the following information, where relevant, must be provided:

— chemical name in accordance with IUPAC and CA nomenclature,

— ISO common name or proposed common name if available,

— CAS number, EC (Einecs or ELINCS) number, and CIPAC number if available,

— molecular and structural formula,

— molecular mass, and

— maximum content in g/kg.

Where the manufacturing process is such that impurities and by-products which are particularly undesirable because of their toxicological, ecotoxicological or environmental properties could be present in the active substance, the content of each such compound must be determined and reported. In such cases, the analytical methods used and the limits of determination, which must be sufficiently low, for each compound of concern, must be reported. Additionally the following information, where relevant, must be provided:

— chemical name in accordance with IUPAC and CA nomenclature,

— ISO common name or proposed common name if available,

— CAS number, EC (Einecs or ELINCS) number, and CIPAC number if available,

— molecular and structural formula,

— molecular mass, and

— maximum content in g/kg.

Where the information provided relates to a pilot plant production system, the information required must again be provided once industrial scale production methods and procedures have stabilised, if production changes result in a changed specification of purity.

Where the information provided does not fully identify a component, viz. condensates, detailed information on the composition must be provided for each such component.

The trade name of components added to the active substance, prior to manufacture of formulated product, to preserve stability and facilitate ease of handling, where they are used, must also be provided. Additionally the following information, where relevant, must be provided for such additives:

— chemical name in accordance with IUPAC and CA nomenclature,

— ISO common name or proposed common name if available,

— CAS number, EC (Einecs or ELINCS) number, and CIPAC number if available,

— molecular and structural formula,

— molecular mass, and

— maximum content in g/kg.

For added components, other than active substance and other than impurities resulting from the manufacturing process, the function of the component (additive) must be given:

— antifoaming agent,

— antifreeze,

— binder,

— stabiliser,

— buffer,

— dispersing agent,

— other (specify).

1.11. Analytical profile of batches

Representative samples of the active substance must be analysed for content of pure active substance, inactive isomers, impurities and additives, as appropriate. The analytical results reported must include quantitative data, in terms of g/kg content, for all components present in quantities of more than 1 g/kg and typically shall account for at least 98 % of the material analysed. The actual content of components which are particularly undesirable because of their toxicological, ecotoxicological or environmental properties, must be determined and reported. Data reported must include the results of the analysis of individual samples and a summary of that data, to show the minimum or maximum and typical content of each relevant component, as appropriate.

Where an active substance is produced in different plants this information must be provided for each of the plants separately.

In addition, where available and relevant, samples of the active substance produced in laboratory scale or pilot production systems, must be analysed, if such material was used in generating toxicological or ecotoxicological data.

2. Physical and chemical properties of the active substance

(i) The information provided, must describe the physical and chemical properties of active substances and together with relevant information, must serve to characterise them. In particular, the information provided must permit: — physical, chemical, and technical hazards associated with active substances, to be identified, — classification of active substance as to hazard, — appropriate restrictions and conditions to be associated with approvals, and — appropriate hazard and precautionary statements to be specified. The information and data referred to are required for all active substances, except where otherwise specified.

(ii) The information provided, taken together with that provided for relevant preparations, must permit the physical, chemical and technical hazards associated with preparations, to be identified, permit preparations to be classified, and permit establishment that preparations can be used without unnecessary difficulty, and be such that exposure of man, animals, and the environment is minimised, taking account of manner of use.

(iii) The extent to which active substances of which approval is sought, comply with relevant FAO specifications, must be stated. Divergences from FAO specifications must be described in detail, and justified.

(iv) In certain specified instances, tests must be conducted using purified active substance of stated specification. In such cases the principles of the method(s) of purification must be reported. The purity of such test material, which must be as high as can be achieved using the best available technology, must be reported. A reasoned justification must be provided in cases where the degree of purity achieved is less than 980 g/kg. Such justification must demonstrate that all technically feasible and reasonable possibilities for the production of the pure active substance have been exhausted.

2.1. Melting point and boiling point

2.1.1. The melting point or where appropriate the freezing or solidification point of purified active substance must be determined and reported in accordance with method A 1 of Regulation (EC) No 440/2008. Measurements shall be taken up to 360 °C.

2.1.2. Where appropriate the boiling point of purified active substances must be determined and reported in accordance with method A 2 of Regulation (EC) No 440/2008. Measurements shall be taken up to 360 °C.

2.1.3. Where melting point and/or boiling point cannot be determined because of decomposition or sublimation, the temperature at which decomposition or sublimation occurs, must be reported.

2.2. Relative density

In the case of active substances which are liquids or solids, the relative density of the purified active substance must be determined and reported in accordance with method A 3 of Regulation (EC) No 440/2008.

2.3. Vapour pressure (in Pa), volatility (e.g. Henry’s law constant)

2.3.1. The vapour pressure of purified active substance must be reported in accordance with method A 4 of Regulation (EC) No 440/2008. Where vapour pressure is less than 10–5 Pa, the vapour pressure at 20 or 25 °C may be estimated by a vapour pressure curve.

2.3.2. In the case of active substances which are solids or liquids, volatility (Henry’s law constant) of purified active substance must be determined or calculated from its water solubility and vapour pressure and be reported (in Pa × m3 × mol–1).

2.4. Appearance (physical state, colour and odour; if known)

2.4.1. A description of both the colour, if any, and the physical state of both the active substance as manufactured and purified active substance, must be provided.

2.4.2. A description of any odour associated with the active substance as manufactured and purified active substance, noted when handling the materials in laboratories or production plants, must be reported.

2.5. Spectra (UV/VIS, IR, NMR, MS), molecular extinction at relevant wavelengths

2.5.1. The following spectra including a table of signal characteristica needed for interpretation must be determined and reported: Ultraviolet/Visible (UV/VIS), infrared (R), nuclear magnetic resonance (NMR), and mass spectra (MS) of purified active substance and molecular extinction at relevant wavelengths, must be determined and reported.

The wavelengths at which UV/visible molecular extinction occurs are to be determined and reported and must include where appropriate a wavelength at the highest absorption value above 290 nm.

In the case of active substances which are resolved optical isomers their optical purity must be measured and reported.

2.5.2. The UV/visible absorption spectra, IR, NMR and MS spectra, where necessary for the identification of the impurities considered to be of toxicological, ecotoxicological or environmental significance must be determined and reported.

2.6. Solubility in water including effect of pH (4 to 10) on solubility

The water solubility of purified active substances under atmospheric pressure must be determined and reported in accordance with method A 6 of Regulation (EC) No 440/2008. These water solubility determinations must be made in the neutral range (i.e. in distilled water in equilibrium with atmospheric carbon dioxide). Where the active substance is capable of forming ions, determinations must also be made in the acidic range (pH 4 to 6) and in the alkaline range (pH 8 to 10), and be reported. Where the stability of the active substance in aqueous media is such that water solubility cannot be determined, a justification based on test data must be provided.

2.7. Solubility in organic solvents

The solubility of the active substances as manufactured in the following organic solvents at 15 to 25 °C must be determined and reported if less than 250 g/kg; the temperature applied must be specified:

— Aliphatic hydrocarbon: preferably n-heptane,

— Aromatic hydrocarbon: preferably xylene,

— Halogenated hydrocarbon: preferably 1,2-dichlorethane,

— Alcohol: preferably methanol or isopropyl alcohol,

— Ketone: preferably acetone,

— Ester: preferably ethyl acetate.

If for a particular active substance, one or more of these solvents are unsuitable (e.g. reacts with test material), alternative solvents can be used instead. In such cases, the choices made must be justified in terms of their structure and polarity.

2.8. Partition coefficient n-octanol/water including effect of pH (4 to 10)

The n-octanol/water partition coefficient of purified active substance must be determined and reported in accordance with method A 8 of Regulation (EC) No 440/2008. The effect of pH (4 to 10) must be investigated when the substance is acidic or basic as defined by its pKa value (< 12 for acids, > 2 for bases).

2.9. Stability in water, hydrolysis rate, photochemical degradation, quantum yield and identity of breakdown product(s), dissociation constant including effect of pH (4 to 9)

2.9.1. The hydrolysis rate of purified active substances (usually radiolabelled active substance, > 95 % purity), for each of the pH values 4, 7 and 9, under sterile conditions, in the absence of light, must be determined and reported in accordance with method C 7 of Regulation (EC) No 440/2008. For substances with a low rate of hydrolysis, the rate can be determined at 50 °C, or another appropriate temperature.

If degradation is observed at 50 °C, degradation rate at another temperature must be determined, and an Arrhenius plot must be constructed to permit an estimate to be made of hydrolysis at 20 °C. The identity of hydrolysis products formed and the rate constantly observed, must be reported. The estimated DT50 value must also be reported.

2.9.2. For compounds with a molar (decadic) absorption coefficient (ε) > 10 (1 × mol–1 × cm–1) at a wavelength λ ≥ 290 nm, direct phototransformation in purified (e.g. distilled) water at 20 to 25 °C, of purified active substance usually radio labelled using artificial light under sterile conditions, if necessary using a solubiliser, must be determined and reported. Sensitisers such as acetone must not be used as a cosolvent or solubiliser. The light source must simulate sunlight and be equipped with filters to exclude radiation at wavelengths λ < 290 nm. The identity of breakdown products formed which at any time during the study are present in quantities ≥ 10 % of the active substance added, a mass balance to account for at least 90 % of the applied radioactivity, as well as photochemical halflife must be reported.

2.9.3. Where necessary to investigate direct phototrans-formation, the quantum yield of direct photodegradation in water must be determined and reported, together with calculations to estimate theoretical lifetime of the active substance in the top layer of aqueous systems and the real lifetime of the substance.

The method is described in the FAO Revised Guidelines on Environmental Criteria for the Registration of Pesticides (6).

2.9.4. Where dissociation in water occurs, the dissociation constant(s) (pKa values) of the purified active substance must be determined and reported in accordance with OECD Test Guideline 112. The identity of the dissociated species formed, based on theoretical considerations, must be reported. If the active substance is a salt, the pKa value of the active principle must be given.

2.10. Stability in air, photochemical degradation, identity of breakdown product(s)

An estimation of the photochemical oxidative degradation (indirect phototransformation) of the active substance, must be submitted.

2.11. Flammability including auto-flammability

2.11.1. The flammability of active substances as manufactured, which are solids, gases, or are substances which evolve highly flammable gases, must be determined and reported in accordance with method A 10, A 11 or A 12 of Regulation (EC) No 440/2008 as appropriate.

2.11.2. The auto-flammability of active substances as manufactured must be determined and reported in accordance with method A 15 or A 16 of Regulation (EC) No 440/2008 as appropriate, and/or, where necessary in accordance with the UN-Bowes-Cameron-Cage-Test (UN-Recommendations on the Transport of Dangerous Goods, Chapter 14, No 14.3.4).

2.12. Flash point

The flash point of active substances as manufactured with a melting point below 40 °C, must be determined and reported in accordance with method A 9 of Regulation (EC) No 440/2008; only closed cup methods shall be used.

2.13. Explosive properties

The explosive properties of active substances as manufactured, must be determined and reported in accordance with method A 14 of Regulation (EC) No 440/2008 where necessary.

2.14. Surface tension

The surface tension has to be determined and reported in accordance with method A 5 of Regulation (EC) No 440/2008.

2.15. Oxidising properties

The oxidising properties of active substances as manufactured, must be determined and reported in accordance with method A 17 of Regulation (EC) No 440/2008, except where examination of its structural formula, establishes beyond reasonable doubt that the active substance is incapable of reacting exothermically with a combustible material. In such cases, it is sufficient to provide that information as justification for not determining the oxidising properties of the active substance.

3. Further information on the active substance

(i) The information provided must describe the intended purposes for which preparations containing the active substance are used, or are to be used and the dose and manner of their use or proposed use.

(ii) The information provided must specify the normal methods and precautions to be followed, in the handling, storage and transport of the active substance.

(iii) The studies, data and information submitted, together with other relevant studies, data and information, must both specify and justify the methods and precautions to be followed in the event of fire. The possible products of combustion in the event of fire shall be estimated, based on the chemical structure and the chemical and physical properties of the active substance.

(iv) The studies, data and information submitted, together with other relevant studies, data and information, must demonstrate the suitability of measures proposed for use in emergency situations.

(v) The information and data referred to are required for all active substances, except where otherwise specified.

3.1. Function, e.g. fungicide, herbicide, insecticide, repellent, growth regulator

The function must be specified from among the following:

— acaricide

— bactericide

— fungicide

— herbicide

— insecticide

— molluscicide

— nematicide

— plant growth regulator

— repellent

— rodenticide

— semio-chemicals

— talpicide

— viricide

— other (must be specified).

3.2. Effects on harmful organisms, e.g. contact poison, inhalation poison, stomach poison, fungitoxic, etc. systematic or not in plants

3.2.1. The nature of the effects on harmful organisms must be stated:

— contact action

— stomach action

— inhalation action

— fungitoxic action

— fungistatic action

— desiccant

— reproduction inhibitor

— other (must be specified).

3.2.2. It must be stated whether or not the active substance is translocated in plants and where relevant whether such translocation is apoplastic, symplastic or both.

3.3. Field of use envisaged, e.g. field, protected crops, storage of plant products, home gardening

The field(s) of use, existing and proposed, for preparations containing the active substance must be specified from among the following:

— Field use, such as agriculture, horticulture, forestry and viticulture

— Protected crops

— Amenity

— Weed control on non-cultivated areas

— Home gardening

— House plants

— Plant products storage practice

— Other (specify).

3.4. Harmful organisms controlled and crops or products protected or treated

3.4.1. Details of existing and the intended use in terms of crops, groups of crops, plants, or plant products treated and where relevant protected, must be provided.

3.4.2. Where relevant, details of harmful organisms against which protection is afforded, must be provided.

3.4.3. Where relevant, effects achieved e.g. sprout suppression, retardation of ripening, reduction in stem length, enhanced fertilisation, etc. must be reported.

3.5. Mode of action

3.5.1. To the extent that is has elucidated, a statement must be provided as to the mode of action of the active substance in terms, where relevant, of the biochemical and physiological mechanism(s) and biochemical pathway(s) involved. Where available, the results of relevant experimental studies must be reported.

3.5.2. Where it is known that to exert its intended effect, the active substance must be converted to a metabolite or degradation product following application or use of preparations containing it, the following information, cross referenced to and drawing on information provided in the context of paragraphs 5.6, 5.11, 6.1, 6.2, 6.7, 7.1, 7.2 and 9, where relevant, must be provided for active metabolite or degradation product:

— chemical name in accordance with IUPAC and CA nomenclature,

— ISO common name or proposed common name,

— CAS EC-number EC (Einecs or ELINCS) number, and CIPAC number if available,

— empirical and structural formula, and

— molecular mass.

3.5.3. Available information relating to the formation of active metabolites and degradation products must be provided, to include:

— the processes, mechanisms and reactions involved,

— kinetic and other data concerning the rate of conversion and if known the rate limiting step,

— environmental and other factors effecting the rate and extent of conversion.

3.6. Information on the occurrence or possible occurrence of the development of resistance and appropriate management strategies

Where available, information on possible occurrence of the development of resistance or cross-resistance must be provided.

3.7. Recommended methods and precautions concerning handling, storage, transport or fire

A safety data sheet pursuant to Article 31 of Regulation (EC) No 1907/2006 of the European Parliament and of the Council (7) must be provided for all active substances.

3.8. Procedures for destruction or decontamination

3.8.1. Controlled incineration

In many cases the preferred or sole means to safely dispose of active substances, contaminated materials, or contaminated packaging, is through controlled incineration in a licensed incinerator.

Where the content of halogens of the active substance is greater than 60 %, the pyrolytic behaviour of the active substance under controlled conditions (including where relevant supply of oxygen and defined residence time), at 800 °C and the content of polyhalogenated dibenzo-p-dioxins and dibenzo-furans in the products of pyrolysis must be reported. The application must provide detailed instructions for safe disposal.

3.8.2. Others

Other methods to dispose of the active substance, contaminated packaging and contaminated materials, where proposed, must be fully described. Data must be provided for such methods, to establish their effectiveness and safety.

3.9. Emergency measures in case of an accident

Procedures for the decontamination of water in case of an accident must be provided.

4. Analytical methods

Introduction

The provisions of this Section only cover analytical methods required for post-registration control and monitoring purposes.

For analytical methods used for generation of data as required in this Regulation or for other purposes the applicant has to provide a justification for the method used; where necessary separate guidance will be developed for such methods on the basis of the same requirements as defined for methods for post-registration control and monitoring purposes.

Descriptions of methods must be provided and include details of equipment, materials and conditions used.

As far as practicable these methods must employ the simplest approach, involve the minimum cost, and require commonly available equipment.

For this Section the following applies:

Impurities, metabolites, relevant metabolites as defined in Regulation (EC) No 1107/2009 Relevant impurities Impurities of toxicological and/or ecotoxicological or environmental concern Significant impurities Impurities with a content of ≥ 1 g/kg in the active substance as manufactured

On request the following samples must be provided:

(i) analytical standards of the pure active substance;

(ii) samples of the active substance as manufactured;

(iii) analytical standards of relevant metabolites and all other components included in the residue definition;

(iv) if available, samples of reference substances for the relevant impurities.

4.1. Methods for the analysis of the active substance as manufactured

For this point the following definitions apply:

(i) Specificity

Specificity is the ability of a method to distinguish between the analyte being measured and other substances.

(ii) Linearity

Linearity is defined as the ability of the method, within a given range, to obtain an acceptable linear correlation between the results and the concentration of analyte in samples.

(iii) Accuracy

The accuracy of a method is defined as the degree to which the determined value of analyte in a sample corresponds to the accepted reference value (for example ISO 5725).

(iv) Precision

Precision is defined as the closeness of agreement between independent test results obtained under prescribed conditions.

Repeatability: Precision under repeatability conditions, i.e. conditions where independent test results are obtained with the same method on identical test material in the same laboratory by the same operator using the same equipment within short intervals of time.

The reproducibility is not required for the active substance as manufactured (for definition of reproducibility see ISO 5725).

4.1.1. Methods, which must be described in full, must be provided for the determination of pure active substance in the active substance as manufactured as specified in the dossier submitted in support of approval. The applicability of existing Cipac methods must be reported.

4.1.2. Methods must also be provided for the determination of significant and/or relevant impurities and additives (e.g. stabilisers) in the active substance as manufactured.

4.1.3. Specificity, linearity, accuracy and repeatability

4.1.3.1. Specificity of methods submitted must be demonstrated and reported. In addition, the extent of interference by other substances present in the active substance as manufactured (e.g. isomers, impurities or additives), must be determined.

While interferences due to other components may be identified as systematic errors in the assessment of the accuracy of methods proposed for the determination of pure active substance in the active substance as manufactured, an explanation must be provided for any interference occurring which contributes more than ± 3 % to the total quantity determined. The degree of interference for methods for the determination of impurities must also be demonstrated.

4.1.3.2. The linearity of proposed methods over an appropriate range must be determined and reported. For the determination of pure active substance, the calibration range must extend (by at least 20 %) the highest and lowest nominal content of the analyte in relevant analytical solutions. Duplicate calibration determinations must be made at three or more concentrations. Alternatively, five concentrations, each as single measurements, are acceptable. Reports submitted must include the equation of the calibration line and the correlation coefficient and representative and properly labelled documentation from the analysis, e.g. chromatograms.

4.1.3.3. Accuracy is required for methods for the determination of pure active substance and significant and/or relevant impurities in the active substance as manufactured.

4.1.3.4. For the repeatability in the determination of the pure active substance in principle a minimum of five determinations must be made. The relative standard deviation (% RSD) must be reported. Outliers identified through an appropriate method (e.g. Dixons or Grubbs test), may be discarded. Where outliers have been discarded, that fact must be clearly indicated. An explanation as to the reason for the occurrence of individual outliers must be attempted.

4.2. Methods for the determination of residues

The methods must be capable of determining the active substance and/or relevant metabolites. For each method and for each relevant representative matrix, the specificity, precision, recovery, and limit of determination must be experimentally determined and reported.

In principle, residue methods proposed shall be multi-residue methods; a standard multi-residue method must be assessed and reported as to its suitability for residue determination. Where residue methods proposed are not multi-residue methods, or are not compatible with such methods, an alternative method must be proposed. Where this requirement results in an excessive number of methods for individual compounds, a ‘common moiety method’ may be acceptable.

For this Section the following definitions apply:

(i) Specificity

Specificity is the ability of a method to distinguish between the analyte being measured and other substances.

(ii) Precision

Precision is defined as the closeness of agreement between independent test results obtained under prescribed conditions.

Repeatability: Precision under repeatability conditions, i.e. conditions where independent test results are obtained with the same method on identical test material in the same laboratory by the same operator using the same equipment within short intervals of time.

Reproducibility: As the definition of reproducibility in relevant publications (for example, in ISO 5725) is in general not practicable for residue analytical methods, reproducibility in the context of this Regulation is defined as a validation of repeatability of recovery, from representative matrices and at representative levels, by at least one laboratory which is independent from that which initially validated the study (this independent laboratory may be within the same company) (independent laboratory validation).

(iii) Recovery

The percentage of the amount of active substance or relevant metabolite originally added to a sample of the appropriate matrix which contains no detectable level of the analyte.

(iv) Limit of determination

The limit of determination (often referred to as limit of quantification) is defined as the lowest concentration tested, at which an acceptable mean recovery is obtained (normally 70 to 110 % with a relative standard deviation of preferably ≤ 20 %; in certain justified cases lower or higher mean recovery rates as well as higher relative standard deviations may be acceptable).

4.2.1. Residues in and/or on plants, plant products, foodstuffs (of plant and animal origin), feedingstuffs

Methods submitted must be suitable for the determination of all components included in the residue definition as submitted in accordance with points 6.1 and 6.2 in order to enable Member States to determine compliance with established MRL’s or to determine dislodgeable residues.

The specificity of the methods must enable all components included in the residue definition to be determined, using an additional confirmatory method if appropriate.

The repeatability must be determined and reported. The replicate analytical portions for test can be prepared from a common field treated sample, containing incurred residues. Alternatively the replicate analytical portions for test can be prepared from a common untreated sample with aliquots fortified at the required level(s).

The results from an independent laboratory validation must be reported.

The limit of determination including the individual and mean recovery must be determined and reported. The overall relative standard deviation, as well as the relative standard deviation for each fortification level must be experimentally determined and reported.

4.2.2. Residues in soil

Methods for analysis of soil for parent compound and/or relevant metabolites must be submitted.

The specificity of the methods must enable the parent compound and/or relevant metabolites to be determined, using an additional confirmatory method if appropriate.

The repeatability, recovery and the limit of determination including the individual and mean recovery must be determined and reported. The overall relative standard deviation, as well as the relative standard deviation for each fortification level must be experimentally determined and reported.

The proposed limit of determination must not exceed a concentration which is of concern with regard to exposure of non-target organisms or because of phytotoxic effects. Normally the proposed limit of determination shall not exceed 0,05 mg/kg.

4.2.3. Residues in water (including drinking water, ground water and surface water)

Methods for analysis in water for parent compound and/or relevant metabolites must be submitted.

The specificity of the methods must enable the parent compound and/or relevant metabolites to be determined, using an additional confirmatory method if appropriate.

The repeatability, recovery and the limit of determination including the individual and mean recovery must be determined and reported. The overall relative standard deviation, as well as the relative standard deviation for each fortification level must be experimentally determined and reported.

For drinking water the proposed limit of determination must not exceed 0,1 μg/l. For surface water the proposed limit of determination must not exceed a concentration which has an impact on non-target organisms deemed to be unacceptable in accordance with the requirements of the Annex to Commission Regulation (EU) No 546/2011 (8).

4.2.4. Residues in air

Methods for the analysis in air of the active substance and/or relevant metabolites formed during or shortly after application must be submitted unless it can be justified that exposure of operators, workers or bystanders is not likely to occur.

The specificity of the methods must enable the parent compound and/or relevant metabolites to be determined, using an additional confirmatory method if appropriate.

The repeatability, recovery and the limit of determination including the individual and mean recovery must be determined and reported. The overall relative standard deviation, as well as the relative standard deviation for each fortification level must be experimentally determined and reported.

The proposed limit of determination must take into account relevant health based limit values or relevant exposure levels.

4.2.5. Residues in body fluids and tissues

Where an active substance is classified as toxic or highly toxic appropriate analytical methods must be submitted.

The specificity of the methods must enable the parent compound and/or relevant metabolites to be determined, using an additional confirmatory method if appropriate.

The repeatability, recovery and the limit of determination including the individual and mean recovery must be determined and reported. The overall relative standard deviation, as well as the relative standard deviation for each fortification level must be experimentally determined and reported.

5. Toxicological and metabolism studies

Introduction

(i) The information provided, taken together with that provided for one or more preparations containing the active substance, must be sufficient to permit an evaluation to be made as to the risks for man, associated with the handling and use of plant protection products containing the active substance, and the risk for man arising from residual traces remaining in food and water. In addition, the information provided must be sufficient to: — permit a decision to be made as to whether, or not, the active substance can be approved, — specify appropriate conditions or restrictions to be associated with any approval, — classify the active substance as to hazard, — establish a relevant acceptable daily intake (ADI) level for man, — establish acceptable operator exposure level(s) (AOEL), — specify the pictograms, signal words, and relevant hazard and precautionary statements for the protection of man, animals and the environment to be included in packaging (containers), — identify relevant first aid measures as well as appropriate diagnostic and therapeutic measures to be followed in the event of poisoning in man, and — permit an evaluation to be made as to the nature and extent of the risks for man, animals (species normally fed and kept or consumed by man) and of the risks for other non-target vertebrate species.

(ii) There is a need to investigate and report all potentially adverse effects found during routine toxicological investigations (including effects on organs and special systems such as immunotoxicity and neurotoxicity) and to undertake and report such additional studies which may be necessary to investigate the probable mechanism involved, to establish Noaels (no observed adverse effect levels), and to assess the significance of these effects. All available biological data and information which are relevant to the assessment of the toxicological profile of the substance tested, must be reported.

(iii) In the context of the influence that impurities can have on toxicological behaviour, it is essential that for each study submitted, a detailed description (specification) of the material used, as mentioned under point 1.11 of Part A be provided. Tests shall be conducted using active substance of that specification to be used in the manufacture of preparations to be authorised, except where radiolabelled material is required or permitted.

(iv) Where studies are conducted using an active substance produced in the laboratory or in a pilot plant production system, the studies must be repeated using the active substance as manufactured, unless it can be justified that the test material used is essentially the same, for the purposes of toxicological testing and assessment. In cases of uncertainty, appropriate bridging studies must be submitted to serve as a basis for a decision as to the possible need for repetition of the studies.

(v) In the case of studies in which dosing extends over a period, dosing shall preferably be done using a single batch of active substance if stability permits.

(vi) For all studies actual achieved dose in mg/kg body weight, as well as in other convenient units, must be reported. Where dosing via the diet is utilised the test compound must be distributed uniformly in the diet.

(vii) Where, as a result of metabolism or other processes in or on treated plants, or as a result of processing of treated products, the terminal residue (to which consumers or workers as defined in the Annex to Regulation (EU) No 545/2011 point 7.2.3 of Part A will be exposed) contains a substance which is not the active substance itself and is not identified as a metabolite in mammals, it will be necessary to carry out toxicity studies on these components of the terminal residue unless it can be demonstrated that consumer or worker exposure to these substances does not constitute a relevant risk to health. Toxicokinetic and metabolism studies relating to metabolites and degradation products shall only be conducted if toxicity findings of the metabolite cannot be evaluated by the available results relating to the active substance.

(viii) The way of administration of the test substance depends on the main exposure routes. In cases where exposure is mainly by the gas phase, it can be more appropriate to perform inhalation studies instead of oral studies.

5.1. Studies on absorption, distribution, excretion and metabolism in mammals

Quite limited data, as described below and restricted to one test species (normally the rat) may be all that is required in this area. These data can provide information useful in the design and interpretation of subsequent toxicity tests. However, it must be remembered that information on interspecies differences may be crucial in extrapolation of animal data to man and information on percutaneous penetration, absorption, distribution, excretion and metabolism may be useful in operator risk assessments. It is not possible to specify detailed data requirements in all areas, since the exact requirements will be dependent upon the results obtained for each particular test substance.

Aim of the test:

The tests shall provide sufficient data to permit:

— an evaluation of the rate and extent of absorption,

— an evaluation of the tissue distribution and the rate and extent of excretion of the test substance and the relevant metabolites,

— the identification of metabolites and the metabolic pathway.

The effect of dose level on these parameters and whether the results are different after single versus repeated doses, shall also be investigated.

Circumstances in which required

A single dose toxicokinetic study in rats (oral route of administration) in at least two dose levels as well as a repeated dose toxicokinetic study in rats (oral route of administration) at a single dose level, must be conducted and reported. It may be necessary in some cases to perform additional studies on another species (such as goat or chicken).

Test guideline

Regulation (EC) No 440/2008, Method B 36, Toxicokinetics.

5.2. Acute toxicity

The studies, data and information to be provided and evaluated must be sufficient to permit the identification of effects following a single exposure to the active substance, and in particular to establish, or indicate:

— the toxicity of the active substance,

— the time course and characteristics of the effects with full details of behavioural changes and possible gross pathological findings at post-mortem,

— where possible mode of toxic action, and

— the relative hazard associated with the different routes of exposure.

While the emphasis must be on estimating the toxicity ranges involved, the information generated must also permit the active substance to be classified in accordance with Regulation (EC) No 1272/2008. The information generated through acute toxicity testing is of particular value in assessing hazards likely to arise in accident situations.

5.2.1. Oral

Circumstances in which required

The acute oral toxicity of the active substance must always be reported.

Test guideline

The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 1 bis or B 1 ter.

5.2.2. Percutaneous

Circumstances in which required

The acute percutaneous toxicity of the active substance must always be reported.

Test guideline

Both local and systemic effects must be investigated. The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, method B 3.

5.2.3. Inhalation

Circumstances in which required

The inhalation toxicity of the active substance must be reported where the active substance is:

— a gas or liquified gas,

— is to be used as a fumigant,

— is to be included in a smoke generating, aerosol or vapour releasing preparation,

— is to be used with fogging equipment,

— has a vapour pressure > 1 × 10–2 Pa and is to be included in preparations to be used in enclosed spaces such as warehouses or glasshouses,

— is to be included in preparations which are powders containing a significant proportion of particles of diameter < 50 μm (> 1 % on a weight basis), or

— is to be included in preparations to be applied in a manner which generates a significant proportion of particles or droplets of diameter < 50 μm (> 1 % on a weight basis).

Test guideline

The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 2.

5.2.4. Skin irritation

Aim of the test

The test will provide the potential of skin irritancy of the active substance including the potential reversibility of the effects observed.

Circumstances in which required

The skin irritancy of the active substance must be determined except where it is likely, as indicated in the test guideline, that severe skin effects may be produced or that effects can be excluded.

Test guideline

The acute skin irritation must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 4.

5.2.5. Eye irritation

Aim of test

The test will provide the potential of eye irritancy of the active substance including the potential reversibility of the effects observed.

Circumstances in which required

Eye irritation tests must be conducted except where it is likely, as indicated in the test guideline, that severe effects on the eyes may be produced.

Test guidelines

The acute eye irritation must be determined in accordance with the Annex to Regulation (EC) No 440/2008, Method B 5.

5.2.6. Skin sensitisation

Aim of test

The test will provide sufficient information to assess the potential of the active substance to provoke skin sensitisation reactions.

Circumstances in which required

The test must always be carried out except where the substance is a known sensitiser.

Test guideline

The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 6.

5.3. Short-term toxicity

Short-term toxicity studies must be designed to provide information as to the amount of the active substance that can be tolerated without toxic effects under the conditions of the study. Such studies provide useful data on the risks for those handling and using preparations containing the active substance. In particular, short-term studies provide an essential insight into possible cumulative actions of the active substance and the risks to workers who may be intensively exposed. In addition short-term studies provide information useful in the design of chronic toxicity studies.

The studies, data and information to be provided and evaluated, must be sufficient to permit the identification of effects following repeated exposure to the active substance, and in particular to further establish, or indicate:

— the relationship between dose and adverse effects,

— toxicity of the active substance including where possible the Noael,

— target organs, where relevant,

— the time course and characteristics of poisoning with full details of behavioural changes and possible pathological findings at post-mortem,

— specific toxic effects and pathological changes produced,

— where relevant the persistence and reversibility of certain toxic effects observed, following discontinuation of dosing,

— where possible, the mode of toxic action, and

— the relative hazard associated with the different routes of exposure.

5.3.1. Oral 28-day study

Circumstances in which required

Although it is not mandatory to perform 28-day short-term studies, they can be useful as range finding tests. Where conducted, they must be reported, since the results could be of particular value in the identification of adaptive responses which can be masked in chronic toxicity studies.

Test guideline

The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 7.

5.3.2. Oral 90-day study

Circumstances in which required

The short-term oral toxicity (90 day) of the active substance to both rat and dog, must always be reported. Where there is evidence that the dog is significantly more sensitive and where such data are likely to be of value in extrapolating results obtained to man, a 12-month toxicity study in dogs must be conducted and reported.

Test guidelines

The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Methods B 26 and B 27, sub-chronic oral toxicity test repeated dose 90.

5.3.3. Other routes

Circumstances in which required

For the assessment of operator exposure additional percutaneous studies may be useful.

For volatile substances (vapour pressure > 10–2 Pascal) expert judgment is required to decide whether the short-term studies have to be performed by oral or inhalation exposure.

Test guidelines

— 28-day dermal: the Annex to Regulation (EC) No 440/2008, Method B 9, repeated dose toxicity (dermal),

— 90-day dermal: the Annex to Regulation (EC) No 440/2008 Method B 28, sub-chronic dermal toxicity study,

— 28-day inhalation: the Annex to Regulation (EC) No 440/2008, Method B 8, repeated dose toxicity (inhalation),

— 90-day inhalation: the Annex to Regulation (EC) No 440/2008, Method B 29, sub-chronique inhalation toxicity study.

5.4. Genotoxicity testing

Aim of the test

These studies are of value in:

— the prediction of genotoxic potential

— the early identification of genotoxic carcinogens

— the elucidation of the mechanism of action of some carcinogens.

To avoid responses that are artefacts of the test system, excessively toxic doses must not be used in either in vitro or in vivo assays for mutagenicity. This approach shall be regarded as general guidance. It is important that a flexible approach is adopted, with selection of further tests which are dependent upon interpretation of results at each stage.

5.4.1. In vitro studies

Circumstances in which required

In vitro mutagenicity tests (bacterial assay for gene mutation, test for clastogenicity in mammalian cells and test for gene mutation in mammalian cells) must always be performed.

Test guidelines

Acceptable test guidelines are:

— Annex to Regulation (EC) No 440/2008, Method B 13/14 — reverse mutation test using bacteria,

— Annex to Regulation (EC) No 440/2008, Method B 10 — in vitro mammalian chromosome aberration test,

— Annex to Regulation (EC) No 440/2008, Method B 17 — in vitro mammalian cell gene mutation test.

5.4.2. In vivo studies in somatic cells

Circumstances in which required

If all the results of the in vitro studies are negative further testing must be done with consideration of other relevant information available (including toxicokinetic, toxicodynamic and physico-chemical data and data on analogous substances). The test can be an in vivo study or an in vitro study using a different metabolising system from that/those previously used.

If the in vitro cytogenetic test is positive, an in vivo test using somatic cells (metaphase analysis in rodent bone marrow or micronucleus test in rodents) must be conducted.

If either of the in vitro gene mutation tests are positive, an in vivo test to investigate unscheduled DNA synthesis or a mouse spot test must be conducted.

Test guidelines

The following test guidelines are acceptable:

— Annex to Regulation (EC) No 440/2008, Method B 12 — In vivo mammalian erythrocyte micronucleus test,

— Annex to Regulation (EC) No 440/2008, Method B 24— Mouse spot test,

— Annex to Regulation (EC) No 440/2008, Method B 11 — In vivo Mammalian Bone-Marrow chromosome aberration test.

5.4.3. In vivo studies in germ cells

Circumstances in which required

When any result of an in vivo study in somatic cells is positive, in vivo testing for germ cell effects may be justified. The necessity for conducting these tests will have to be considered on a case by case basis, taking into account information regarding toxicokinetics, use and anticipated exposure. Suitable tests would need to examine interaction with DNA (such as the dominant lethal assay), to look at the potential for inherited effects and possibly make a quantitative assessment of heritable effects. It is recognised that in view of their complexity, the use of quantitative studies would require strong justification.

5.5. Long-term toxicity and carcinogenicity

Aim of the test

The long-term studies conducted and reported, taken together with other relevant data and information on the active substance, must be sufficient to permit the identification of effects, following repeated exposure to the active substance, and in particular must be sufficient to:

— identify adverse effects resulting from exposure to the active substance,

— identify target organs, where relevant,

— establish the dose-response relationship,

— identify changes in toxic signs and manifestations observed, and

— establish the Noael.

Similarly, the carcinogenicity studies taken together with other relevant data and information on the active substance, must be sufficient to permit the hazards for humans, following repeated exposure to the active substance, to be assessed, and in particular must be sufficient:

— to identify carcinogenic effects resulting from exposure to the active substance,

— to establish the species and organ specificity of tumours induced,

— to establish the dose-response relationship, and

— for non-genotoxic carcinogens, to identify the maximum dose eliciting no adverse effect (threshold dose).

Circumstances in which required

The long-term toxicity and carcinogenicity of all active substances must be determined. If in exceptional circumstances, it is claimed that such testing is unnecessary, that claim must be fully justified, viz. toxicokinetic data demonstrates that absorption of the active substance does not occur from the gut, through the skin or via the pulmonary system.

Test conditions

A long-term oral toxicity and carcinogenicity study (2 years) of the active substance must be conducted using the rat as test species; these studies can be combined.

A carcinogenicity study of the active substance must be conducted using the mouse as test species.

Where a non-genotoxic mechanism for carcinogenicity is suggested, a well argued case, supported with relevant experimental data, including that necessary to elucidate the possible mechanism involved, must be provided.

While the standard reference points for treatment responses are concurrent control data, historical control data, may be helpful in the interpretation of particular carcinogenicity studies. Where submitted, historical control data shall be from the same species and strain, maintained under similar conditions and shall be from contemporaneous studies. The information on historical control data provided must include:

— identification of species and strain, name of the supplier, and specific colony identification, if the supplier has more than one geographical location,

— name of the laboratory and the dates when the study was performed,

— description of the general conditions under which animals were maintained, including the type or brand of diet and, where possible, the amount consumed,

— approximate age, in days, of the control animals at the beginning of the study and at the time of killing or death,

— description of the control group mortality pattern observed during or at the end of the study, and other pertinent observations (e.g. diseases, infections),

— name of the laboratory and the examining scientists responsible for gathering and interpreting the pathological data from the study, and

— a statement of the nature of the tumours that may have been combined to produce any of the incidence data.

The doses tested, including the highest dose tested, must be selected on the basis of the results of short-term testing and where available at the time of planning the studies concerned, on the basis of metabolism and toxicokinetic data. The highest dose level in the carcinogenicity study shall elicit signs of minimal toxicity such as slight depression in body-weight gain (less than 10 %), without causing tissue necrosis or metabolic saturation and without substantially altering normal lifespan due to effects other than tumours. If the long-term toxicity study is carried out separately, the highest dose level shall elicit definite signs of toxicity without causing excessive lethality. Higher doses, causing excessive toxicity are not considered relevant to evaluations to be made.

In the collection of data and compilation of reports, incidence of benign and malignant tumours must not be combined, unless there is clear evidence of benign tumours becoming malignant with time. Similarly, dissimilar, un-associated tumours, whether benign or malignant, occurring in the same organ, must not be combined, for reporting purposes. In the interests of avoiding confusion, terminology such as that developed by the American Society of Toxicologic Pathologists (9), or the Hannover Tumour Registry (RENI) shall be used in the nomenclature and reporting of tumours. The system used must be identified.

It is essential that biological material selected for histopathological examination includes material selected to provide further information on lesions identified during gross pathological examination. Where relevant to the elucidation of mechanism of action and available, special histological (staining) techniques, histochemical techniques and electron microscopic examinations, must be conducted and reported.

Test guideline

The studies must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 30 Chronic toxicity test, Method B 32 Carcinogenicity test or Method B 33 combined chronic toxicity/carcinogenicity test.

5.6. Reproductive toxicity

The adverse reproductive effects are of two main types:

— impairment of male or female fertility, and

— impacts on the normal development of progeny (developmental toxicity).

Possible effects on all aspects of reproductive physiology in both males and females, as well as possible effects on pre-natal and post-natal development, must be investigated and reported. If in exceptional circumstances, it is claimed that such testing is unnecessary, that claim must be fully justified.

While the standard reference point for treatment responses are concurrent control data, historical control data may be helpful in the interpretation of particular reproductive studies. Where submitted, historical control data shall be from the same species and strain, maintained under similar conditions and shall be from contemporaneous studies. The information on historical control data provided must include:

— identification of species and strain, name of the supplier, and specific colony identification, if the supplier has more than one geographical location,

— name of the laboratory and the dates when the study was performed,

— description of the general conditions under which animals were maintained, including the type or brand of diet and, where possible, the amount consumed,

— approximate age, in days, of the control animals at the beginning of the study and at the time of killing or death,

— description of the control group mortality pattern observed during or at the end of the study, and other pertinent observations (e.g. diseases, infections), and

— name of the laboratory and the examining scientist responsible for gathering and interpreting the toxicological data from the study.

5.6.1. Multi-generation studies

Aim of the test

The studies reported, taken together with other relevant data and information on the active substance, must be sufficient to permit the identification of effects for reproduction, following repeated exposure to the active substance, and in particular must be sufficient:

— to identify direct and indirect effects on reproduction resulting from exposure to the active substance,

— to identify any enhancement of general toxic effects (noted during short-term and chronic toxicity testing),

— to establish the dose-response relationship,

— to identify changes in toxic signs and manifestations observed, and

— to establish the Noael.

Circumstances in which required

A reproduction toxicity study in rats over at least two generations must always be reported.

Test guideline

The tests must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 35, two-generation reproduction toxicity study. In addition organ weight of reproductive organs must be reported.

Supplementary studies

Where necessary for a better interpretation of the effects on reproduction and as far as this information is not yet available it could be necessary to perform supplementary studies in order to provide the following information:

— separate male and female studies,

— three segment designs,

— dominant lethal assay for male fertility,

— cross-matings of treated males with untreated females and vice versa,

— effects on spermatogenesis,

— effects on oogenesis,

— sperm motility, mobility and morphology, and

— investigation of hormonal activity.

5.6.2. Developmental toxicity studies

Aim of the test

The studies reported, taken together with other relevant data and information on the active substance, must be sufficient to permit effects on embryonic and foetal development, following repeated exposure to the active substance, to be assessed, and in particular must be sufficient:

— to identify direct and indirect effects on embryonic and foetal development resulting from exposure to the active substance,

— to identify any maternal toxicity,

— to establish the relationship between observed responses and dose in both dam and offspring,

— to identify changes in toxic signs and manifestations observed, and

— to establish the Noael.

Furthermore, the tests will give additional information on any enhancement of general toxic effects of pregnant animals.

Circumstances in which required

The tests must always be carried out.

Test conditions

Developmental toxicity must be determined both to rat and rabbit by the oral route. Malformations and variations shall be reported separately. A glossary of terminology and diagnostic principles for malformations and variations must be given in the report.

Test guideline

The tests must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 31, prenatal developmental toxicity study.

5.7. Delayed neurotoxicity studies

Aim of the test

The test shall provide sufficient data to evaluate if the active substance could provoke delayed neurotoxicity after acute exposure.

Circumstances in which required

These studies have to be performed for substances of similar or related structures to those capable of inducing delayed neurotoxicity such as organophosphates.

Test guidelines

The test must be carried out in accordance with OECD Guideline 418.

5.8. Other toxicological studies

5.8.1. Toxicity studies of metabolites as referred to in the introduction, point (vii)

Supplementary studies, where they relate to substances other than the active substance, are not a routine requirement.

Decisions as to the need for supplementary studies must be made on a case by case basis.

5.8.2. Supplementary studies on the active substance

In certain cases it can be necessary to carry out supplementary studies to further clarify observed effects. These studies could include:

— studies on absorption, distribution, excretion and metabolism,

— studies on the neurotoxic potential,

— studies on the immunotoxicological potential,

— studies on other routes of administration.

Decisions as to the need for supplementary studies must be made on a case by case basis, taking into account the results of the available toxicological and metabolism studies and the most important exposure routes.

Studies required must be designed on an individual basis, in the light of the particular parameters to be investigated and the objectives to be achieved.

5.9. Medical data

Where available, and without prejudice to the provisions of Article 10 of Council Directive 98/24/EC (10), practical data and information relevant to the recognition of the symptoms of poisoning, and on the effectiveness of first aid and therapeutic measures have to be submitted. More specific references to the investigation for antidotal pharmacology or safety pharmacology using animals shall be provided. Where relevant, the effectiveness of potential antagonists to poisoning, shall be investigated and reported.

Data and information relevant to the effects of human exposure, where available and of the necessary quality, are of particular value, in confirming the validity of extrapolations made and conclusions reached with respect to target organs, dose-response relationships, and the reversibility of toxic effects. Such data can be generated following accidental or occupational exposure.

5.9.1. Medicinal surveillance on manufacturing plant personnel

Reports of occupational health surveillance programmes, supported with detailed information on the design of the programme, on exposure to the active substance and exposure to other chemicals, must be submitted. Such reports shall, where feasible, include data relevant to the mechanism of action of the active substance. These reports shall, where available, include data from persons exposed in manufacturing plants or after application of the active substance (e.g. in efficacy trials).

Available information on the sensitisation including allergenic response of workers and others exposed to the active substance, must be provided, and include where relevant details of any incidence of hypersensitivity. The information provided shall include details of frequency, level and duration of exposure, symptoms observed and other relevant clinical information.

5.9.2. Direct observation, e.g. clinical cases and poisoning incidents

Available reports from the open literature, relating to clinical cases and poisoning incidents, where they are from refereed journals or official reports, must be submitted together with reports of any follow-up studies undertaken. Such reports shall contain complete descriptions of the nature, level and duration of exposure, as well as the clinical symptoms observed, first aid and therapeutic measures applied and measurements and observations made. Summary and abstract information is not of value.

Where supported with the necessary level of detail, such documentation can be of particular value in confirming the validity of extrapolations from animal data to man and in identifying unexpected adverse effects which are specific to humans.

5.9.3. Observations on exposure of the general population and epidemiological studies if appropriate

Where available, and supported with data on levels and duration of exposure, and conducted in accordance with recognised standards (11), epidemiological studies are of particular value and must be submitted.

5.9.4. Diagnosis of poisoning (determination of active substance, metabolites), specific signs of poisoning, clinical tests

A detailed description of the clinical signs and symptoms of poisoning, including the early signs and symptoms and full details of clinical tests useful for diagnostic purposes, where available, must be provided and include full details of the time courses involved relevant to the ingestion, dermal exposure or inhalation of varying amounts of the active substance.

5.9.5. Proposed treatment: first aid measures, antidotes, medical treatment

The first aid measures to be used in the event of poisoning (actual and suspected) and in the event of contamination of eyes must be provided.

Therapeutic regimes for use in the event of poisoning or contamination of eyes, including where available the use of antidotes, must be described in full. Information based on practical experience, where it exists and is available, in other cases on theoretical grounds, as to the effectiveness of alternative treatment regimes, where relevant, must be provided. Contraindications associated with particular regimes, particularly those relating to ‘general medical problems’ and conditions, must be described.

5.9.6. Expected effects of poisoning

Where known, the expected effects and the duration of these effects following poisoning must be described and include the impact of:

— the type, level and duration of exposure, or ingestion, and

— varying time periods between exposure, or ingestion, and commencement of treatment.

5.10. Summary of mammalian toxicity and overall evaluation

A summary of all data and information provided under paragraphs 5.1 through 5.10, must be submitted, and include a detailed and critical assessment of those data in the context of relevant evaluative and decision making criteria and guidelines, with particular reference to the risks for man and animals that may or do arise, and the extent, quality and reliability of the data base.

Where relevant, in the light of findings with respect to the analytical profile of batches of the active substance (point 1.11) and any bridging studies conducted (point (iv) of the introduction to Section 5), the relevance of the data as submitted to the assessment of the toxicological profile of the active substance as manufactured, must be argued.

On the basis of an assessment of the data base, and the relevant decision making criteria and guidelines, justifications must be submitted for the Noaels proposed for each relevant study.

On the basis of these data scientifically reasoned proposals for the establishment of ADI and AOEL(s) for the active substance must be submitted.

6. Residues in or on treated products, food and feed

Introduction

(i) The information provided, taken together with that provided for one or more preparations containing the active substance, must be sufficient to permit an evaluation to be made as to the risks for man, arising from residues of the active substance and relevant metabolites, degradation and reaction products remaining in food. In addition, the information provided must be sufficient to: — permit a decision to be made as to whether, or not, the active substance can be approved, — specify appropriate conditions or restrictions to be associated with any approval.

(ii) A detailed description (specification) of the material used, as provided under point 1.11 must be provided.

(iii) Studies shall be performed in accordance with the EU Guidelines for the generation of data concerning residues (12).

(iv) Where relevant, data shall be analysed using appropriate statistical methods. Full details of the statistical analysis shall be reported.

(v) Stability of residues during storage. If may be necessary to perform studies on the stability of residues during storage. Provided samples are frozen within generally 24 hours after sampling and unless a compound is otherwise known to be volatile or labile, data are not normally required for samples extracted and analysed within 30 days from sampling (6 months in the case of radio-labelled material). Studies with non-radio-labelled substances shall be carried out with representative substrates and preferably on samples from treated crops or animals with incurred residues. Alternatively, if this is not possible, aliquots of prepared control samples shall be spiked with a known amount of chemical before storage under normal storage conditions. Where the degradation during storage is significant (more than 30 %) it may be necessary to change the storage conditions or not to store the samples prior to analysis and repeat any studies where the unsatisfactory storage conditions were used. Detailed information with respect to the sample preparation and storage conditions (temperature and duration) of samples and extracts must be submitted. Storage stability data using sample extracts will also be required unless samples are analysed within 24 hours of extraction.

6.1. Metabolism, distribution and expression of residue in plants

Aim of the tests

The objectives of these studies are:

— to provide an estimate of total terminal residues in the relevant portion of crops at harvest following treatment as proposed,

— to identify the major components of the total terminal residue,

— to indicate the distribution of residues between relevant crops parts,

— to quantify the major components of the residue and to establish the efficiency of extraction procedures for these components,

— to decide on the definition and expression of a residue.

Circumstances in which required

These studies must always be performed unless it can be justified that no residues will remain on plants/plant products which are used as food or feeding stuffs.

Test conditions

Metabolism studies have to involve crops or categories of crops in which plant protection products containing the active substance in question would be used. If a wide range of uses in different crop categories or in the category fruits is envisaged, studies have to be carried out on at least three crops unless it can be justified that a different metabolism is unlikely to occur. In cases where use is envisaged in different categories of crops, the studies must be representative for the relevant categories. For this purpose crops can be considered as falling into one of five categories: root vegetables, leafy crops, fruits, pulses and oilseeds, cereals. If studies are available for crops from three of these categories and the results indicate that the route of degradation is similar in all three categories then it is unlikely that any more studies will be needed unless it could be expected that a different metabolism will occur. The metabolism studies have also to take into account the different properties of the active substance and the intended method of application.

An evaluation of the results from different studies has to be submitted on the point and path of uptake (e.g. via leaves or roots), and on the distribution of residues between relevant parts of the crop at harvest (with particular emphasis on edible parts for man or animals). If the active substance or relevant metabolites are not taken up by the crop, this must be explained. Information on the mode of action and the physico-chemical properties of the active substance may be helpful in assessing trial data.

6.2. Metabolism, distribution and expression of residue in livestock

Aim of tests

The objectives of these studies are:

— to identify the major components of the total terminal residue in edible animal products,

— to quantify the rate of degradation and excretion of the total residue in certain animal products (milk or eggs) and excreta,

— to indicate the distribution of residues between relevant edible animal products,

— to quantify the major components of the residue and to show the efficiency of extraction procedures for these components,

— to generate data from which a decision on the need for livestock feeding studies as provided for in point 6.4 can be made,

— to decide on the definition and expression of a residue.

Circumstances in which required

Metabolism studies on animals, such as lactating ruminants (e.g. goat or cow) or laying poultry, are only required when pesticide use may lead to significant residues in livestock feed (≥ 0,1 mg/kg of the total diet as received, except special cases e.g. active substances which accumulate). Where it becomes apparent that metabolic pathways differ significantly in the rat as compared to ruminants a pig study must be conducted unless the expected intake by pigs is not significant.

6.3. Residue trials

Aim of the tests

The objectives of these studies are:

— to quantify the highest likely residue levels in treated crops at harvest or outloading from store following the proposed good agricultural practice (GAP), and

— to determine, when appropriate, the rate of decline of plant protection product deposits.

Circumstances in which required

These studies must always be performed where the plant protection product will be applied to plants/plant products which are used as food or feeding stuffs or where residues from soil or other substrates can be taken up by such plants except where extrapolation from adequate data on another crop is possible.

Residue trial data shall be submitted in the dossier for those uses of plant protection products for which authorisation is sought at the moment of the submission of a dossier for approval of the active substance.

Test conditions

Supervised trials shall correspond to proposed critical GAP. The test conditions must take into account the highest residues which may reasonably arise (e.g. maximum number of proposed applications, use of the maximum envisaged quantity, shortest pre-harvest intervals, withholding periods or storage periods) but which remain representative of the realistic worst case conditions in which the active substance would be used.

Sufficient data must be generated and submitted to confirm that patterns determined hold for the regions and the range of conditions, likely to be encountered in the regions concerned for which its use is to be recommended.

When establishing a supervised trial programme, normally factors such as climatic differences existing between production areas, differences in production methods (e.g. outdoor versus glasshouse uses), seasons of production, type of formulations, etc. shall be taken into account.

In general, for a comparable set of conditions, trials shall be carried out over a minimum of two growing seasons. All exceptions shall be fully justified.

The precise number of trials necessary is difficult to determine in advance of a preliminary evaluation of the trial results. Minimum data requirements only apply where comparability can be established between production areas, e.g. concerning climate, methods and growing seasons of production, etc. Assuming all other variables (climate, etc.) are comparable, a minimum of eight trials representative of the proposed growing area is required for major crops. For minor crops normally four trials representative of the proposed growing area are required.

Due to the inherently higher level of homogeneity in residues arising from post-harvest treatments or protected crops, trials from one growing season will be acceptable. For post-harvest treatments, in principle a minimum of four trials are required, carried out preferably at different locations with different cultivars. A set of trials has to be carried out for each application method and store type unless the worst case residue situation can be clearly identified.

The number of studies per growing season to be performed can be reduced if it can be justified that the residue levels in plants/plant products will be lower than the limit of determination.

Where a significant part of the consumable crop is present at the time of application, half of the supervised residue trials reported shall include data to show the effect of time on the level of residue present (residue decline studies) unless it can be justified that the consumable crop is not affected by the application of the plant protection product under the proposed conditions of use.

6.4. Livestock feeding studies

Aim of the tests

The objective of these studies is to determine the residue in products of animal origin which will result from residues in feedingstuffs or fodder crops.

Circumstances in which required

Feeding studies are only required:

— when significant residues (≥ 0,1 mg/kg of the total diet as received, except special cases, such as active substances which accumulate) occur in crops or part of the crop (e.g. trimmings, waste) fed to animals, and

— when metabolism studies indicate that significant residues (0,01 mg/kg or above the limit of determination if this would be higher than 0,01 mg/kg) may occur in any edible animal tissue taking into account the residue levels in potential feedingstuffs obtained at the 1 × dose rate.

Where appropriate separate feeding studies for lactating ruminant and/or laying poultry shall be submitted. Where it appears from the metabolism studies submitted in accordance with point 6.2 that metabolic pathways differ significantly in the pig as compared to ruminants, a pig feeding study must be conducted unless the expected intake by pigs is not significant.

Test conditions

In general, the feed is administered in three dosages (expected residue level, three to five times, and 10 times the expected residue level). When setting the 1 × dose, a theoretical feed ration must be compiled.

6.5. Effects of industrial processing and/or household preparations

Circumstances in which required

The decision as to whether it is necessary to carry out processing studies will depend on:

— the importance of a processed product in the human or animal diet,

— the level of residue in the plant or plant product to be processed,

— the physico-chemical properties of the active substance or relevant metabolites, and

— the possibility that degradation products of toxicological significance may be found after processing of the plant or plant product.

Processing studies are not normally necessary if no significant or no analytically determinable residues occur in the plant or plant product which would be processed, or if the total theoretical maximum daily intake (TMDI) is less than 10 % of the ADI. In addition, processing studies are not normally required for plants or plant products mostly eaten raw except for those with inedible portions such as citrus, banana or kiwi fruit where data on the distribution of the residue in peel/pulp may be required.

‘Significant residues’ generally refer to residues above 0,1 mg/kg. If the pesticide concerned has a high acute toxicity and/or a low ADI, consideration must be given to conducting processing studies for determinable residues below 0,1 mg/kg.

Studies on the effects on the nature of the residue are not normally required where only simple physical operations, not involving a change in temperature of the plant or the plant product, are involved such as washing, trimming or pressing.

6.5.1. Effects on the nature of the residue

Aim of the tests

The objective of these studies is to establish whether or not breakdown or reaction products arise from residues in the raw products during processing which may require a separate risk assessment.

Test conditions

Depending upon the level and chemical nature of the residue in the raw commodity, a set of representative hydrolysis situations (simulating the relevant processing operations) shall be investigated, where appropriate. The effects of process other than hydrolysis may also have to be investigated, where the properties of the active substance or metabolites indicate that toxicologically significant degradation products may occur as a result of these processes. The studies are normally conducted with a radio-labelled form of the active substance.

6.5.2. Effects on the residue levels

Aim of the tests

The main objectives of these studies are:

— to determine the quantitative distribution of residues in the various intermediate and end products, and to estimate transfer factors,

— to enable a more realistic estimate to be made of dietary intake of residues.

Test conditions

Processing studies shall represent household processing and/or actual industrial processes.

In the first instance it is usually only necessary to carry out a core set of ‘balance studies’ representative of the common processes relevant to the plants or plant products containing significant residues. Justification shall be given for the selection made of these representative process(es). The technology to be used in processing studies shall always correspond as closely as possible to the actual conditions that are normally used in practice. A balance sheet shall be made in which the mass balance of residues in all intermediate and end products is investigated. In drawing up such a balance sheet any concentrations or reductions in residues in individual products can be recognised and the corresponding transfer factors can also be determined.

If the processed plant products play an important part in the diet, and if the ‘balance study’ indicates that a significant transfer of residue into the processed products could occur, then three ‘follow-up studies’ to determine residue concentration or dilution factors must be carried out.

6.6. Residues in succeeding crops

Aim of the test

The objective of these studies is to permit an evaluation of possible residues in succeeding crops.

Circumstances in which required

Where data generated in accordance with point 7.1 of this Annex or point 9.1 of the Annex to Regulation (EU) No 545/2011, shows that significant residues (> 10 % of the applied active substance as a total of unchanged active substance and its relevant metabolites or degradation products) remain in soil or in plant materials, such as straw or organic material up to sowing or planting time of possible succeeding crops, and which could lead to residues above the limit of determination in succeeding crops at harvest, consideration shall be given to the residue situation. This shall include consideration of the nature of the residue in the succeeding crops and involve at least a theoretical estimation of the level of these residues. If the likelihood of residues in succeeding crops can not be excluded, metabolism and distribution studies shall be carried out, if necessary followed by field trials.

Test conditions

If a theoretical estimation of residues in succeeding crops is done, full details and a justification shall be given.

Metabolism and distribution studies and field trials, if necessary, shall be carried out on representative crops chosen to represent normal agricultural practice.

6.7. Proposed maximum residue levels (MRLs) and residue definition

A full justification for the proposed MRLs must be provided, including, where relevant, full details of the statistical analysis used.

When judging which compounds are to be included in the residue definition, account has to be taken of the toxicological significance of the compounds, amounts likely to be present and the practicality of the analytical methods proposed for post-registration control and monitoring purposes.

6.8. Proposed pre-harvest intervals for envisaged uses, or withholding periods or storage periods, in the case of post-harvest uses

A full justification for the proposals must be provided.

6.9. Estimation of the potential and actual exposure through diet and other means

Consideration will be given to the calculation of a realistic prediction of dietary intake. This may be done in a step-wise fashion leading to an increasingly realistic prediction of intake. Where relevant, other sources of exposure such as residues arising from the use of medicines or veterinary drugs have to be taken into account.

6.10. Summary and evaluation of residue behaviour

A summary and evaluation of all data presented in this Section shall be carried out in accordance with the guidance given by the competent authorities of the Member States concerning the format of such summaries and evaluations. It shall include a detailed and critical assessment of those data in the context of relevant evaluative and decision-making criteria and guidelines, with particular reference to the risks for man and animals that may or do arise, and the extent, quality and reliability of the data base.

In particular, the toxicological significance of any non-mammalian metabolites must be addressed.

A schematic diagram shall be prepared of the metabolic pathway in plants and animals with a brief explanation of the distribution and chemical changes involved.

7. Fate and behaviour in the environment

Introduction

(i) The information provided, taken together with that for one or more preparations containing the active substance, must be sufficient to permit an assessment of the fate and behaviour of the active substance in the environment, and of the non-target species likely to be at risk from exposure to the active substance, its metabolites, degradation and reaction products, where they are of toxicological or environmental significance.

(ii) In particular, the information provided for the active substance, together with other relevant information, and that provided for one or more preparations containing it shall be sufficient to: — decide whether, or not, the active substance can be approved, — specify appropriate conditions or restrictions to be associated with any approval, — classify the active substance as to hazard, — specify the pictograms, signal words and relevant hazard and precautionary statements for the protection of the environment, which are to be included on packaging (containers), — predict the distribution, fate, and behaviour in the environment of the active substance and relevant metabolites, degradation and reaction products as well as the times courses involved, — identify non-target species and populations for which hazards arise because of potential exposure, and — identify measures necessary to minimise contamination of the environment and impact on non-target species.

(iii) A detailed description (specification) of the material used, as provided for under point 1.11 must be provided. Where testing is done using active substance the material used shall be of that specification that will be used in the manufacture of preparations to be authorised except where radio-labelled material is used. Where studies are conducted using active substance produced in the laboratory or in a pilot plant production system, the studies must be repeated using active substance as manufactured, unless it can be justified that the test material used is essentially the same for the purposes of environmental testing and assessment.

(iv) Where radio-labelled test material is used, radio-labels shall be positioned at sites (one or more as necessary), to facilitate elucidation of metabolic and degradative pathways and to facilitate investigation of the distribution of the active substance and of its metabolite, reaction and degradation products in the environment.

(v) It may be necessary to conduct separate studies for metabolites, degradation or reaction products, where these products can constitute a relevant risk to non-target organisms or to the quality of water, soil and air and where their effects cannot be evaluated by the available results relating to the active substance. Before such studies are performed the information from the Sections 5 and 6 has to be taken into account.

(vi) Where relevant, tests shall be designed and data analysed using appropriate statistical methods. Full details of the statistical analysis shall be reported (e.g. all point estimates shall be given with confidence intervals, exact p-values shall be given rather than stating significant/non-significant).

7.1. Fate and behaviour in soil

All relevant information on the type and the properties of the soil used in the studies, including pH, organic carbon content, cation exchange capacity, particle size distribution and water holding capacity, at pF = 0 and pF = 2,5 has to be reported in accordance with relevant ISO or other international standards.

The microbial biomass of soils used for laboratory degradation studies must be determined just prior to the commencement and at the end of the study.

It is recommended to use as much as possible the same soils throughout all laboratory soil studies.

The soils used for degradation or mobility studies must be selected such that they are representative of the range of soils typical of the v