The information provided must be sufficient to identify with precision each active substance, to define it in terms of its specification and to characterize it as to its nature. The information and data referred to, unless otherwise specified, are required for all active substances.
The name and address of the applicant (permanent Community address) 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 inclusion in Annex I 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.
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 inclusion of the active substances in Annex I, there are changes in the location or number of manufacturers, the information required must again be notified to the Commission and the Member States.
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.
The Chemical name as given in Annex I to Directive 67/548/EEC, or, if not included in this Directive, in accordance with both IUPAC and CA nomenclature, must be provided.
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.
Chemical Abstracts, EEC (EINECS or ELINCS), and CIPAC numbers, where they exist, must be reported.
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.
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 stabilized.
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 stabilized, if production changes result in a changed specification of purity.
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 according to IUPAC and CA nomenclature,
ISO common name or proposed common name if available,
CAS number, EEC (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 according to IUPAC and CA nomenclature,
ISO common name or proposed common name if available,
CAS number, EEC (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 stabilized, 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 according to IUPAC and CA nomenclature,
ISO common name or proposed common name if available,
CAS number, EEC (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,
other (specify),
buffer,
dispersing agent,
stabilizer.
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 should 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 analyzed, if such material was used in generating toxicological or ecotoxicological data.
Textual Amendments
The information provided, must describe the physical and chemical properties of active substances and together with relevant information, must serve to characterize 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 harzard,
appropriate restrictions and conditions to be associated with inclusions in Annex I to be selected, and
appropriate risk and safety phrases to be specified.
The information and data referred to are required for all active substances, except where otherwise specified.
The information provided, taken together with that provided for relevant preparations, must permit the physical, chemical 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 minimized, taking account of manner of use.
The extent to wich active substances of which inclusion in Annex I is sought, comply with relevant FAO specifications, must be stated. Divergences from FAO specifications must be described in detail, and justified.
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.
In the case of active substances which are liquids or solids, the relative density of the purified active substance must be determined and reported according to EEC method A 3.
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.
The water solubility of purified active substances under atmospheric pressure must be determined and reported according to EEC method A 6. 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.
The solubility of the active substances as manufactured in the following organic solvents at 15 to 25 o 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 is unsuitable (e.g. reacts with test material), alternative solvents can be used instead. In such cases, choices made must be justified in terms of their structure and polarity.
The n-octanol/water partition coefficient of purified active substance must be determined and reported according to EEC method A 8. 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).
If degradation is observed at 50 o 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 o C. The identity of hydrolysis products formed and the rate constantly observed, must be reported. The estimated DT 50 value must also be reported.
The method is described in the FAO Revised Guidelines on Environmental Criteria for the Registration of Pesticides.
An estimation of the photochemical oxidative degradation (indirect hototransformation) of the active substance, must be submitted.
The flash point of active substances as manufactured with a melting point below 40 o C, must be determined and reported according to EEC method A 9; only closed cup methods should be used.
The explosive properties of active substances as manufactured, must be determined and reported according to EEC method A 14 where necessary.
The surface tension has to be determined and reported according to EEC method A 5.
The oxidizing properties of active substances as manufactured, must be determined and reported according to EEC method A 17, except where examination of its sturctural 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 oxidizing properties of the substance.
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.
The information provided must specify the normal methods and precautions to be followed, in the handling, storage and transport of the active substance.
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 should be estimated, based on the chemical structure and the chemical and physical properties of the active substance.
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.
The informaton and data referred to are reqired for all active substances, except where otherwise specified.
The function must be specified from among the following:
acaricide
bactericide
fungicide
herbicide
insecticide
molluscicide
nematicide
plant growth regulator
repellant
rodenticide
semio-chemicals
talpicide
viricide
other (must be specified)
contact action
stomach action
inhalation action
fungitoxic action
fungistatic action
desiccant
reproduction inhibitor
other (must be specified)
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)
chemical name according to IUPAC and CA nomenclature,
ISO common name or porposed common name,
CAS EEC-number EEC (EINECS or ELINCS) number, and CIPAC number if available,
empirical and structural formula, and
molecular mass.
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.
Where available information on possible occurrence of the development of resistance or cross-resistance must be provided.
A safety data sheet pursuant to Article 27 of Council Directive 65/548/EEC (2) must be provided for all active substances.
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 o 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.
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.
Procedures for the decontamination of water in case of an accident must be provided.]
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 Directive 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 | Any component other than the pure active substance which is present in the active substance as manufactured (including non-active isomers) originating from the manufacturing process or from degradation during storage, |
| 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, |
| Metabolites | Metabolites include products resulting from degradation or reaction of the active substance, |
| Relevant metabolites | Metabolites of toxicological and/or ecotoxicological or environmental concern. |
On request the following samples must be provided:
analytical standards of the pure active substance;
samples of the active substance as manufactured;
analytical standards of relevant metabolites and all other components included in the residue definition;
if available, samples of reference substances for the relevant impurities.
For this point the following definitions apply:
Specificity
Specificity is the ability of a method to distinguish between the analyte being measured and other substances.
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.
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).
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).
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.
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 should 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:
Specificity
Specificity is the ability of a method to distinguish between the analyte being measured and other substances.
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 Directive 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).
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.
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).
Methods submitted must be suitable for the determination of all components included in the residue definition as submitted according to the provisions of section 6, 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.
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 should not exceed 0,05 mg/kg.
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 recovvery 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 according to the requirements of Annex VI.
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.
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.]
Textual Amendments
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 included in Annex I,
specify appropriate conditions or restrictions to be associated with any inclusion in Annex I,
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 hazard symbols, the indications of danger, and the risk and safety phrases 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.
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 is relevant to the assessment of the toxicological profile of the substance tested, must be reported.
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 section 1 point 11 be provided. Tests should be conducted using active substance of that specification to be used in the manufacture of preparations to be authorized, except where radiolabelled material is required or permitted.
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.
In the case of studies in which dosing extends over a period, dosing should preferably be done using a single batch of active substance if stability permits.
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 utilized the test compound must be distributed uniformly in the diet.
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 Annex III, point 7.2.3 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 should only be conducted if toxicity findings of the metabolite cannot be evaluated by the available results relating to the active substance.
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.
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 dependant upon the results obtained for each particular test substance.
The tests should provide sufficient data to permit:
an evaluation of the rate and extent of absorption,
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 results are different after single versus repeated doses, should also be investigated.
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).
Commission Directive 87/302/EEC of 18 November 1987 adapting to technical progress for the ninth time Council Directive 67/548/EEC on the approximation of laws, regulations and administrative provisions relating to the classification, packaging and labelling of dangerous substances (3) , part B, Toxicokinetics.
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 Council Directive 67/548/EEC. The information generated through acute toxicity testing is of particular value in assessing hazards likely to arise in accident situations.
The acute oral toxicity of the active substance must always be reported.
The test must be carried out in accordance with the Annex to Commission Directive 92/69/EEC of 31 July 1992 adapting to technical progress for the seventeenth time Council Directive 67/548/EEC on the approximation of laws, regulations and administrative provisions relating to the classification, packaging and labelling of dangerous substances (4) , Method B1 or B1 bis.
The acute percutaneous toxicity of the active substance must always be reported.
Both local and systemic effects must be investigated. The test must be carried out in accordance with Directive 92/69/EEC method B3.
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& minus;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 [X1< 50µm] (> 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 < 50µm (> 1 % on a weight basis).
Editorial Information
The test must be carried out in accordance with Directive 92/69/EEC Method B2.
The test will provide the potential of skin irritancy of the active substance including the potential reversibility of the effects observed.
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.
The acute skin irritation must be carried out in accordance with Directive 92/69/EEC Method B4.
The test will provide the potential of eye irritancy of the active substance including the potential reversibility of the effects observed.
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.
The acute eye irritation must be determined in accordance with Directive 92/69/EEC Method B5.
The test will provide sufficient information to assess the potential of the active substance to provoke skin sensitization reactions.
The test must always be carried out except where the substance is a known sensitizer.
The test must be carried out in accordance with Directive 92/69/EEC Method B6.
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.
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.
The test must be carried out in accordance with Directive 92/69/EEC Method B7.
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.
Directive 87/302/EEC, Part B, sub-chronic oral toxicity test.
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.
28-day dermal: Directive 92/69/EEC Method B9,
90-day dermal: Directive 87/302/EEC, Part B, sub-chronic dermal toxicity study,
28-day inhalation: Directive 92/69/EEC Method B8,
90-day inhalation: Directive 87/302/EEC, Part B, sub-chronic inhalation toxicity study.
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 artifacts of the test system, excessively toxic doses must not be used in either in vitro or in vivo assays for mutagenicity. This approach should be regarded as general guidance. It is important that a flexible approach is adopted, with selection of further tests being' dependant upon interpretation of results at each stage.
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.
Acceptable test guidelines are:
Directive 92/69/EEC Method B14 — Salmonella Typhimurium reverse mutation assay
Directive 92/69/EEC Method B10 — in vitro mammalian cytogenetic test
Directive 87/302/EEC, Part B — in vitro mammalian cell gene mutation test
If all the results of the in vitro studies are negative further resting 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 metabolizing 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.
Acceptable test guidelines are:
Directive 92/69/EEC Method B12 — Micronucleus test,
Directive 87/302/EEC Part B — Mouse spot test,
Directive 92/69/EEC Method B11 — In vivo Mammalian Bone-Marrow cytogenetic test, Chromosomal analysis.
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 recognized that in view of their complexity, the use of quantitative studies would require strong justification.
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).
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.
A long-term oral toxicity and carcinogenicity study (two 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 should be from the same species and strain, maintained under similar conditions and should 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 should 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 should 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 American Society of Toxicologic Pathologists (5) , or the Hannover Tumour Registry (RENI) should 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.
The studies must be carried out in accordance with Directive 87/302/EEC, part B, Chronic toxicity test, Carcinogenicity test or combined chronic toxicity/carcinogenicity test.
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 should be from the same species and strain, maintained under similar conditions and should 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.
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.
A reproduction toxicity study in rats over at least two generations must always be reported.
The tests must be carried out in accordance with Directive 87/302/EEC, Part B, two-generation reproduction toxicity test. In addition organ weight of reproductive organs must be reported.
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.
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.
The tests must always be carried out.
Developmental toxicity must be determined both to rat and rabbit by the oral route. Malformations and variations should be reported separately. A glossary of terminology and diagnostic principles for malformations and variations must be given in the report.
The tests must be carried out in accordance with Directive 87/302/EEC, Part B, teratogenicity test — rodent and non-rodent.
The test shall provide sufficient data to evaluate if the active substance could provoke delayed neurotoxicity after acute exposure.
These studies have to be performed for substances of similar or related structures to those capable of inducing delayed neurotoxicity such as organophosphates.
The test must be carried out in accordance with OECD Guideline 418.
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.
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.
Where available, and without prejudice to the provisions of Article 5 of Council Directive 80/1107/EEC of 27 November 1980 on the protection of workers from the risks related to chemical, physical and biological agents at work (6) , 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 should be provided. Where relevant, the effectiveness of potential antagonists to poisoning, should 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.
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 should, 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 sensitization 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 should include details of frequency, level and duration of exposure, symptoms observed and other relevant clinical information.
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 should 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.
Where available, and supported with data on levels and duration of exposure, and conducted in accordance with recognized standards (7) , epidemiological studies are of particular value and must be submitted.
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.
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.
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.
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 (paragraph 1.11) and any bridging studies conducted (paragraphs 5 (iv)), 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.]
Textual Amendments
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 included in Annex I,
specify appropriate conditions or restrictions to be associated with any inclusion in Annex I.
A detailed description (specification) of the material used, as provided under Section 1, point 11 must be provided.
Studies should be performed according to the guidance available on regulatory testing procedures for residues of plant protection products in food (8) .
Where relevant, data should be analyzed using appropriate statistical methods. Full details of the statistical analysis should be reported.
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 (six months in the case of radio-labelled material).
Studies with non-radio-labelled substances should be carried out with representative substratets and preferably on samples from treated crops or animals with incurred residues. Alternatively, if this is not possible, aliquots of prepared control samples should 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 and 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.
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.
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 feedingstuffs.
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.
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.
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.
The objectives of these studies are:
to quantify the highest likely residue levels in treated crops at harvest or outloading from store following the propopsed good agricultural practice (GAP),
and
to determine, when appropriate, the rate of decline of plant protection product deposits.
These studies must always be performed where the plant protection product will be applied to plants/plant products which are used as food or feedingstuffs 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 Annex II dossier for those uses of plant protection products for which authorization is sought at the moment of introduction of a dossier for inclusion of the active substance in Annex I.
Supervised trials should corrspond 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. should be taken into account.
In general, for a comparable set of conditions, trials should be carried out over a minimum of two growing seasons. All exceptions should 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 should 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.
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.
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 should be submitted. Where it appears from the metabolism studies submitted in accordance with the provisions of 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.
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.
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.
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.
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) should 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.
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.
Processing studies should 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 should be given for the selection made of these representative process(es). The technology to be used in processing studies should always correspond as closely as possible to the actual conditions that are normally used in practice. A balance sheet should 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 recognized 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.
The objective of these studies is to permit an evaluation of possible residues in succeeding crops.
Where data generated in accordance with Annex II, Section 7, point 7.1 or Annex III, Section 9, point 9.1, 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 should be given to the residue situation. This should 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 should be carried out, if necessary followed by field trials.
If a theoretical estimation of residues in succeeding crops is done, full details and a justification shall be be given.
Metabolism and distribution studies and field trials, if necessary, shall be carried out on representative crops chosen to represent normal agricultural practice.
A full justification for the propsoed 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.
A full justification for the proposals must be provided.
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 predictions 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 acocunt.
A summary and evaluation of all data presented in this Section should be carried out according to the guidance given by the competent authorities of the Member States concerning the format of such summaries and evaluations. It should 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 reliablity of the data base.
In particular, the toxicological significance of any non-mammalian metabolites must be addressed.
A schematic diagram should be prepared of the metabolic pathway in plants and animals with a brief explanation of the distribution and chemical changes involved.]
Textual Amendments
The information provided, taken together with that for one or more preparations containing the active substance, must be sufficient to pemit 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.
In particular, the information provided for the active substance, together with other relevant information, and that provided for one or more preparations containing it, should be sufficient to:
decide whether, or not, the active substance can be included in Annex I,
specify appropriate conditions or restrictions to be associated with any inclusion in Annex I,
classify the active substance as to hazard;
specify the hazard symbols, the indications of danger, and relevant risk and safety phrases for the protection of the environment, which are to be included on packaging (containers),
predict the disribution, 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 a rise because of potential exposure, and
identify measures necesary to minimize contamination of the environment and impact on non-target species.
A detailed description (specification) of the material used, as provided for under Section 1, point 11 must be provided. Where testing is done using active substance the material used should be of that specification that will be used in the manufacture of preparations to be authorized 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.
Where radio-labelled test material is used, radio-labels should 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.
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.
Where relevant, tests should be designed and data analysed using appropriate statistical methods.
Full details of the statistical anaylsis should be reported (e.g. all point estimates should be given with confidence intervals, exact p-values should be given rather than stating significant/non significant).
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, 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 various Community regions where use exists or is anticipated, and be such that:
they cover a range of organic carbon content, particle size distribution and pH values; and
where on the basis of other information, degradation or mobility are expected to be pH dependent (e.g. solubility and hydrolysis rate — paragraphs 2.7 and 2.8), they cover the following pH ranges:
4,5 to 5,5
6 to 7, and
8 (approximately).
Soils used must, wherever possible, be freshly sampled. If use of stored soils is unavoidable, storage should be properly carried out for a limited time under defined and reported conditions. Soils stored for longer periods of time can only be used for adsorption/desorption studies.
The soil chosen to begin studying should not have extreme characteristics with respect to parameters such as particle size distribution, organic carbon content and pH.
Soils should be collected and handled in accordance with ISO 10381-6 (Soil quality — Sampling — Guidance on the collection, handling and storage of soil for the assessment of microbial processes in the laboratory) . Any deviations must be reported and justified.
Field studies should be carried out in conditions as close to normal agricultural practice as possible on a range of soil types and climatic conditions representative of the area(s) of use. Weather conditions shall be reported in cases where field studies are conducted.
Aim of the tests
The data and information provided, together with other relevant data and information, should be sufficient to:
identify, where feasable, the relative importance of the types of process involved (balance between chemical and biological degradation),
identify the individual components present which at any time account for more than 10 % of the amount of active substance added, including, where feasible, non-extractable residues,
identify where possible also individual components present which account for less than 10 % of the amount of active substance added,
establish the relative proportions of the components present (mass balance), and
permit the soil residue of concern and to which non-target species are or may be exposed, to be defined.
Where a reference is made to non-extractable residues these are defined as chemical species originating from pesticides used according to good agricultural practice that cannot be extracted by methods which do not significantly change the chemical nature of these residues. These non-extractable residues are not considered to include fragments through metabolic pathways leading to natural products.
Circumstances in which required
The degradation pathway or pathways must always be reported except where the nature and manner of use of preparations containing the active substance, preclude soil contamination such as uses on stored products or wound healing treatments for trees.
Test conditions
The degradation pathway or pathways must be reported for one soil.
Results obtained must be presented in the form of schematic drawings showing the pathways involved, and in the form of balance sheets which show the distribution of radio-label as a function of time, as between:
active substance,
CO 2 ,
volatile compounds other than CO 2 ,
individual identified transformation products,
extractable substances not identified, and
non-extractable residues in soil.
The investigation of degradation pathways must include all feasible steps to characterise and quantify non-extractable residues formed after 100 days when exceeding 70 % of the applied dose of the active substance. The techniques and methodologies applied are best selected on a case-by-case basis. A justification must be provided where the compounds involved are not characterized.
The duration of the study is normally 120 days, except where after a shorter period the levels of non-extractable residues and CO 2 are such that they can be extrapolated in a reliable way to 100 days.
Test guideline
Setac — Procedures for assessing the environmental fate and ecotoxicity of pesticides (9) .
Anaerobic degradation
Circumstances in which required
An anaerobic degradation study must be reported unless it can be justified that exposure of the plant protection products containing the active substance to anaerobic conditions is unlikely to occur.
Test conditions and test guideline
The same provisions as provided for under the corresponding paragraph of point 7.1.1.1.1 apply.
Soil photolysis
Circumstances in which required
A soil photolysis study must be reported unless it can be justified that deposition of the active substance at the soil surface is unlikely to occur.
Test guideline
Setac — Procedures for assessing the Environmental fate and ecotoxicity of pesticides.
Aim of the tests
The soil degradation studies should provide best possible estimates of the time taken for degradation of 50 % and 90 % (DT 50lab and DT 90lab ), of the active substance, and of relevant metabolites, degradation and reaction products under laboratory conditions.
Aerobic degradation
Circumstances in which required
The rate of degradation in soil must always be reported, except where the nature and manner of use of plant protection products containing the active substance preclude soil contamination such as uses on stored products or wound healing treatments for trees.
Test conditions
The rate of aerobic degradation of the active substance in three soil types additional to that referred to in paragraph 7.1.1.1.1. must be reported.
In order to investigate the influence of temperature on degradation, one additional study at 10 o C has to be performed on one of the soils used for the investigation of degradation at 20 o C until a validated Community calculation model for the extrapolation of degradation rates at low temperatures is available.
The duration of the study is normally 120 days except if more than 90 % of the active substance is degraded before that period expires.
Similar studies for three soil types must be reported for all relevant metabolites, degradation and reaction products which occur in soil and which at any time during the studies account for more than 10 % of the amount of active substance added, except where their DT 50 values were able to be determined from the results of the degradation studies with the active substance.
Test guideline
Setac — Procedures for assessing the environmental fate and ecotoxicity of pesticides.
Anaerobic degradation
Circumstances in which required
The rate of anaerobic degradation of the active substance must be reported where an anaerobic study has to be performed according to point 7.1.1.1.2.
Test conditions
The rate of anaerobic degradation of the active substance must be carried out in the soil used in the anaerobic study performed according to point 7.1.1.1.2.
The duration of the study is normally 120 days except if more than 90 % of the active substance is degraded before that period expires.
Similar studies for one soil must be reported for all relevant metobolites, degradation and reaction products which occur in soil and which at any time during the studies account for more than 10 % of the amount of active substance added, except where their DT 50 values were able to be determined from the results of the degradation studies with the active substance.
Test guideline
Setac — Procedures for assessing the environmental fate and ecotoxicity of pesticides.
Soil dissipation studies
Aim of the test
The soil dissipation studies should provide estimates of the time taken for dissipation of 50 % and 90 % (DT 50f and DT 90f ), of the active substance under field conditions. Where relevant, information on relevant metabolites, degradation and reaction products must be reported.
Circumstances in which required
The tests have to be conducted in those conditions where DT 50lab determined at 20 o C and at a moisture content of the soil related to a pF value of 2 to 2,5 (suction pressure) is greater than 60 days.
Where plant protection products containing the active substance are intended to be used in cold climatic conditions, the tests have to be conducted where DT 50lab , determined at 10 o C and at a moisture content of the soil related to a pF value of 2 to 2,5 (suction pressure) is greater than 90 days.
Test conditions
Individual studies on a range of representative soils (normally four different types) must be continued until > 90 % of the amount applied have dissipated. The maximum duration of the studies is 24 months.
Test guideline
SETAC — Procedures for assessing the environmental fate and ecotoxicity of pesticides.
Soil residue studies
Aim of the test
Soil residue studies should provide estimates of the soil residue levels at harvest or at time of cowing or planting succeeding crops.
Circumstances in which required
Soil residue studies must be reported where DT 50lab is greater than one-third of the period between the application and harvest and where absorption by the succeeding crop is possible, except where soil residues at sowing or planting of a succeeding crop can be reliably estimated from the data of the soil dissipation studies or where it can be justified that these residues can not be phytotoxic to or leave unacceptable residues in rotational crops.
Test conditions
Individual studies must be continued until harvest or time of sowing or planting succeeding crops, unless > 90 % of the amount applied have dissipated.
Test guideline
SETAC — Procedures for assessing the environmental fate and ecotoxicity of pesticides.
Soil accumulation studies
Aim of the tests
The tests should provide sufficient data to evaluate the possibility of accumulation of residues of the active substance and of relevant metabolites, degradation and reaction products.
Circumstances in which required
Where on the basis of soil dissipation studies it is established that DT 90f > one year and where repeated application is envisaged, whether in the same growing season or in succeeding years, the possibility of accumulation of residues in soil and the level at which a plateau concentration is achieved must be investigated except where reliable information can be provided by a model calculation or another appropriate assessment.
Test conditions
Long term field studies must be done on two relevant soils and involve multiple applications.
Before performing these studies the applicant shall seek the agreement of the competent authorities on the type of study to be performed.
Aim of the test
The data and information provided, together with other relevant data and information, should be sufficient to establish the absorption coefficient of the active substance and of relevant metabolites, degradation and reaction products.
Circumstances in which required
The studies must always be reported except where the nature and manner of use of preparations containing the active substance, preclude soil contamination such as uses on stored products or wound healing trees.
Testo conditions
Studies on the active substance must be reported for four soil types.
Similar studies, for at least three soil types, must be reported for all relevant metabolites, degradation and reaction products which in soil degradation studies account at any time for more than 10 % of the amount of active substance added.
Test guideline
OECD method 106
Aim of the test
The test should provide sufficient data to evaluate the mobility and leaching potential of the active substance and if possible of relevant metabolities, degradation and reaction products.
Circumstances in which required
Studies in four soils must be carried out where in the absorption and desorption studies provided for under point 7.1.2 it is not possible to obtain reliable absorption coefficient values.
Test guideline
SETAC — Procedures for assessing the environmental fate and ecotoxicity of pesticides.
Aim of the test
The test should provide sufficient data to estimate the mobility and leaching potential of relevant metabolites, degradation and reaction products.
Circumstances in which required
The studies must be performed except:
where the nature and manner of use of preparations containing the active substance, preclude soil contamination such as uses on stored products or wound healing treatments for trees, or
where a separate study for the metabolite, degradation or reaction product in accordance to point 7.1.2 or 7.1.3.1 was performed.
Test conditions
The period(s) of ageing should be determined from inspection of the degradation patterns of active substance and metabolites to ensure that a relevant spectrum of metabolites is present at the time of leaching.
Test guideline
SETAC — Procedures for assessing the environmental fate and ecotoxicity of pesticides.
Aim of the tests
The test should provide data on:
the mobility in soil,
the potential for leaching to ground water,
The potential distribution in soil.
Circumstances in which requried
Expert judgement will be necessary to decide whether lysimeter studies or field leaching studies should be carried out, taking into account the results of degradation and other mobility studies and the predicted environmental concentrations in groundwater (PEC GW ), calculated in accordance with the provisions of Annex III, Section 9. The type and conditions of the study to be conducted should be discussed with the competent authorities.
Test conditions
Great care is necessary in design of both experimental installations and individual studies, to ensure that results obtained can be used for assessment purposes. Studies should cover the realistic worst case situation, taking into account the soil type, climatic conditions, the application rate and the frequency and period of application.
Water percolating from soil columns must be analyzed at suitable intervals, while residues in plant material must be determined at harvest. Residues in the soil profile in at least five layers must be determined on termination of experimental work. Intermediate sampling must be avoided, since removal of plants (except for harvesting according to normal agricultural practice) and soil cross influences the leaching process.
Precipitation, soil and air temperatures have to be recorded at regular intervals (at least on a weekly base).
Lysimeter studies
Test conditions
The minimal depth of the lysimeters should be 100 cm; their maximal depth should be 130 cm. The soil cross must be undisturbed. Soil temperatures must be similar to those pertaining in the field. Where necessary, supplementary irrigation must be provided to ensure optimal plant growth and to ensure that the quantity of infiltration water is similar to that in the regions for which authorization is sought. When during the study the soil has to be disturbed for agricultural reasons it must not be disturbed deeper than 25 cm.
Field leaching studies
Test conditions
Information on the groundwater table in the experimental fields must be submitted. If soil cracking is observed during the study this must be fully described.
Great attention should be given to the number and the location of water collection devices. The placement of these devices in the soil should not result in preferential flow paths.
Test guideline
SETAC — Procedures for assessing the environmental fate and ecotoxicity of pesticides.
Aim of the tests
The information and data provided, taken together with that provided for one or more preparations containing the active substance, and other relevant information, should be sufficient to establish, or permit estimation of:
persistence in water systems (bottom sediment and water, including suspended particles),
the extent to which water, sediment organisms and air are at risk,
potential for contamination of surface water and groundwater.
Aim of the tests
The data and information provided, together with other relevant data and information, should be sufficient to:
identify the relative importance of the types of processes involved (balance between chemical and biological degradation),
where possible, identify the individual components present,
establish the relative proportions of the components present and their distribution as between water, including suspended particles, and sediment, and
permit the residue of concern and to which non-target species are or may be exposed, to be defined.
Circumstances in which required
The test must always be performed for relevant metabolites, degradation and reaction products which account at any time for more than 10 % of the amount of active substance added unless sufficient information on their degradation is available from the test performed in accordance with point 2.9.1.
Test conditions and test guideline
The same provisions as provided under the corresponding paragraphs of point 2.9.1 apply.
Circumstances in which required
The test must always be performed for relevant metabolites, degradation and reaction products which account at any time for more than 10 % of the amount of active substance added unless sufficient information on their degradation is available from the test performed in accordance with points 2.9.2 and 2.9.3.
Test conditions and test guideline
The same provisions as provided under the corresponding paragraphs of points 2.9.2 and 2.9.3 apply.
Circumstances in which required
The test must always be performed unless it is not required under the provisions of Annex VI to Directive 67/548/EEC for the classification of the active substance.
Test guideline
EEC method C4.
Circumstances in which required
The test must be reported unless it can be justified that contamination of surface water will not occur.
Test guideline
SETAC — Procedures for assessing the environmental fate and ecotoxicity of pesticides.
Circumstances in which required
Transformation rates in the saturated zone of active substances and of relevant metabolites, degradation and reaction products can provide useful information on the fate of these substances in the groundwater.
Test conditions
Expert judgement is required to decide whether this information is necessary. Before performing these studies the applicant shall seek the agreement of the competent authorities on the type of study to be performed.
Guidance under development.
In the light of the chemical composition of residues occurring in soil, water or air, resulting from use, or proposed use, of a plant protection product containing the active substance a proposal for the definition of the residue must be submitted, taking account of both the levels found and their toxicological and environmental significance.
Available monitoring data concerning fate and behaviour of the active substance and relevant metabolites, degradation and reaction products must be reported.]
Textual Amendments
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 impact on non-target species (flora and fauna), likely to be at risk from exposure to the active substance, its metabolites, degradation and reaction products, where they are of environmental significance. Impact can result from single, prolonged or repeated exposure and can be reversible or irreversible.
In particular, the information provided for the active substance, together with other relevant information, and that provided for one or more preparations containing it, should be sufficient to:
decide whether, or not, the active substance can be included in Annex I,
specify appropriate conditions or restrictions to be associated with any inclusion in Annex I,
permit an evaluation of short- and long-term risks for non-target species — populations, communities, and processes — as appropriate,
classify the active substance as to hazard,
specify the precautions necessary for the protection of non-target species, and
specify the hazard symbols, the indications of danger, and relevant risk and safety phrases for the protection of the environment, to be mentioned on packaging (containers).
There is a need to report all potentially adverse effects found during routine ecotoxicological investigations and to undertake and report, where required by the competent authorities, such additional studies which may be necessary to investigate the probable mechanisms involved and assess the significance of these effects. All available biological data and information which is relevant to the assessment of the ecotoxicological profile of the active substance must be reported.
The information on fate and behaviour in the environment, generated and submitted in accordance with points 7.1 to 7.4, and on residue levels in plants generated and submitted in accordance with point 6 is central to the assessment of impact on non-target species, in that together with information on the nature of the preparation and its manner of use, it defines the nature and extent of potential exposure. The toxicokinetic and toxicological studies and information submitted in accordance with points 5.1 to 5.8 provide essential information as to toxicity to vertebrate species and the mechanisms involved.
Where relevant, tests should be designed and data analysed using appropriate statistical methods. Full details of the statistical analysis should be reported (e. g. all point estimates should be given with confidence intervals, exact p-values should be given rather than stating significant/non significant).
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 should be of that specification that will be used in the manufacture of preparations to be authorized except where radiolabelled 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 ecotoxicological 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.
In the case of studies in which dosing extends over a period, dosing should preferably be done using a single batch of active substance if stability permits.
Whenever a study implies the use of different doses, the relationship between dose and adverse effect must be reported.
For all feeding studies, average achieved dose must be reported, including where possible the dose in mg/kg body weight. Where dosing via the diet is utilized the test compound must be distributed uniformly in the diet.
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 and where their effects cannot be evaluated by the available results relating to the active substance. Before such studies are performed the information from points 5, 6 and 7 has to be taken into account.
In order to facilitate the assessment of the significance of test results obtained, including the estimation of intrinsic toxicity and the factors affecting toxicity, the same strain (or recorded origin) of each relevant species should, where possible, be used in the various toxicity tests specified.
The test should provide, where possible, LD 50 values, the lethal threshold dose, time courses of response and recovery and the NOEL, and must include relevant gross pathological findings.
The possible effects of the active substance on birds must be investigated except where the active substance is intended solely to be included in preparations for exclusive use in enclosed spaces (e.g. in glasshouses or in food storage practice).
The acute oral toxicity of active substance to a quail species (Japanese quail (Coturnix coturnix japonica) or Bobwhite quail (Colinus virginianus) or to mallard duck (Anas platyrhynchos) must be determined. The highest dose used in tests need not exceed 2 000 mg/kg body weight.
Setac — Procedures for assessing the environmental fate and ecotoxicity of pesticides (10) .
The test should provide the short term dietary toxicity (LC 50 values, lowest lethal concentration (LLC), where possible no observed effect concentrations (NOEC), time courses of response and recovery) and include relevant gross pathological findings.
The dietary (five-day) toxicity of the active substance to birds must always be investigated on one species except where a study in accordance with the provisions of point 8.1.3 is reported. Where its acute oral NOEL is ≤ 500 mg/kg body weight or where the short-term NOEC < 500 mg/kg food the test must be performed on a second species.
The first species to be studied must be either a quail species or mallard duck. If a second species must be tested it should not be related to the first species tested.
The test must be carried out in accordance with OECD Method 205.
The test should provide the subchronic toxicity and reproductive toxicity of the active substance to birds.
The subchronic and reproductive toxicity of the active substance to birds must be investigated, unless it can be justified that continued or repeated exposure of adults, or exposure of nest sites during the breeding season is unlikely to occur.
The test must be carried out in accordance with OECD Method 206.
The data of the tests referred to in points 8.2.1, 8.2.4 and 8.2.6 have to be submitted for every active substance even when it is not expected that plant protection products containing it could reach surface water following the proposed conditions of use. These data are required under the provisions of Annex VI to Directive 67/548/EEC for the classification of the active substance.
Data reported must be supported with analytical data on concentrations of the test substance in the test media.
The test should provide the acute toxicity (LC 50 ), and details of observed effects.
The test must always be carried out.
The acute toxicity of the active substance must be determined for rainbow trout (Oncorhynchus mykiss) and for a warm water fish species. Where tests with metabolites, degradation or reaction products have to be performed the species used must be the more sensitive of the two species tested with the active substance.
The test must be carried out in accordance with the Annex to Commission Directive 92/69/EEC (11) adapting to technical progress for the 17th time Directive 67/548/EEC on the approximation of laws, regulations and administrative provisions relating to the classification and labelling of dangerous, substances, Method C1.
A chronic toxicity study must be carried out unless it can be justified that continued or repeated exposure of fish is unlikely to occur or unless a suitable microsom or mesocosm study is available.
Expert judgment is required to decide which test has to be performed. In particular for active substance for which there are indications of particular concerns (related to the toxicity of the active substance for fish or the potential exposure) the applicant shall seek the agreement of the competent authorities on the type of test to be performed.
A fish early life stage toxicity test might be appropriate where bioconcentration factors (BCF) are between 100 and 1 000 or where EC 50 of the active substance < 0,1 mg/l.
A fish life cycle test might be appropriate in cases where
the bioconcentration factor is greater tan 1 000 and the elimination of the active substance during a depuration phase of 14 days is lower than 95 %,
or
the substance is stable in water or sediment (DT 90 > 100 days).
It is not necessary to perform a chronic toxicity test on juvenile fish when a fish early life stage toxicity test or a fish life cycle test has been performed; it is likewise not necessary to perform a fish early life stage toxicity test when a fish life cycle test has been performed.
The test should provide effects on growth, the threshold level for lethal effects and for observed effects, the NOEC and details of observed effects.
The test must be conducted on juvenile rainbow trout, following exposure of 28 days to the active substance. Data on the effects on growth and behaviour must be generated.
The test should provide effects on development, growth and behaviour, the NOEC and details of observed effects on fish early life stages.
The test must be carried out in accordance with OECD Method 210.
The test will provide effects on reproduction of the parental and the viability of the filial generation.
Before performing these studies the applicant shall seek the agreement of the competent authorities on the type and conditions of the study to be performed.
The test should provide the steady-state bioconcentration factors, uptake rate constants and depuration rate constants, calculated for each test compound, as well as relevant confidence limits.
The bioconcentration potential of active substances, of metabolites and of degradation and reaction products, likely to partition into fatty tissues (such as log p ow ≥ 3 — see point 2.8 or other relevant indications of bioconcentration), must be investigated and be reported, unless it can be justified that exposure leading to bioconcentration is not likely to occur.
The test must be carried out in accordance with OECD Method 305E.
The test should provide the 24 and 48-hour acute toxicity of the active substance, expressed as the median effective concentration (EC 50 ) for immobilization, and where possible the highest concentration causing no immobilization.
The acute toxicity must always be determined for Daphnia (preferably Daphnia magna) . Where plant protection products containing the active substance are intended to be used directly on surface water additional data have to be reported on at least one representative species from each of the following groups: aquatic insects, aqautic crustaceans (on a species not related to Daphnia) and aquatic gastropod molluscs.
The test must be carried out in accordance with Directive 92/69/EEC, Method C2.
The test should provide where possible EC 50 values for effects such as immobilization and reproduction and the highest concentration at which no effect such as on martality or reproduction occurs (NOEC) and details of observed effects.
A test on Daphnia and on at least one representative aquatic insect species and an aquatic gastropod mollusc species must be carried out unless it can be justified that continued or repeated exposure is not likely to occur.
The test with Daphnia must be continued for 21 days.
The test must be carried out in accordance with OECD Method 202, Part II.
The test should procide EC 50 values for growth and growth rate, NOEC values, and details of observed effects.
Possible effects on algal growth of active substances must always be reported.
For herbicides a test on a second species from a different taxonomic group has to be performed.
The test must be carried out in accordance with Directive 92/69/EEC, Method C3.
The test will measure effects on survival and development (including effects on emergence of adults for Chironomus), the relevant EC 50 values and the NOEC values.
Where environmental fate and behaviour data required in point 7 report that an active substance is likely to partition to and persist in aquatic sediments, expert judgement should be used to decide whether an acute or a chronic sediment toxicity test in required. Such expert judgement should take into account whether effects on sediment dwelling invertebrates are likely by comparing the aquatic invertebrate toxicity EC 50 data from points 8.2.4 and 8.2.5 with the predicted levels of the active substances in sediment from data in Annex III, point 9.
Before performing these studies the applicant shall seek the agreement of the competent authorities on the type and conditions of the study to be performed.
A test on aquatic plants has to be performed for herbicides.
Before performing these studies the applicant shall seek the agreement of the competent authorities on the type and conditions of the study to be performed.
The test should provide the acute oral and contact LD 50 value of the active substance.
Potential impact on bees must be investigated, except where preparations containing the active substance are for exclusive use in situations where bees are not likely to be exposed such as:
food storage in enclosed spaces,
non-systemic seed dressings,
non-systemic preparations for application to soil,
non-systemic dipping treatments for transplanted crops and bulbs,
wound sealing and healing treatments,
rodenticidal baits,
use in glasshouses without pollinators.
The test must be carried out in accordance with EPPO Guideline 170.
The test should provide sufficient information to evaluate possible risks from the plant protection product on honeybee larvae.
The test must be carried out when the active substance may act as an insect growth regulator unless it can be justified that it is not likely that bee brood would be exposed to it.
The test must be carried out in accordance with ICPBR Method (e.g. P. A. Oomen, A. de Riujter and J. van der Steen. Method for honeybee brood feeding tests with insect growth-regulating insecticides. EPPO Bulletin, Volume 22, pp 613 to 616, 1992.)
The test should provide sufficient information to evaluate the toxicity (mortality and sublethal effects) of the active substance to selected arthropod species.
Effects on non-target terrestrial arthropods (e.g. predators or parasitoids of harmful organisms) must be investigated. The information obtained for these species can also be used to indicate the potential for toxicity to other non-target species inhabiting the same environment. This information is required for all active substances except where preparations containing the active substance are for exclusive use in situations where non-target arthropods are not exposed such as:
food storage in enclosed spaces,
wound sealing and healing treatments,
rodenticidal baits.
The test must be performed initially in the laboratory on an artificial substrate (i.e. glass plate or quartz sand, as appropriate) unless adverse effects can be clearly predicted from other studies. In these cases, more realistic substrates may be used.
Two sensitive standard species, a parasitoid and predatory mite (e.g. Aphidius rhopalosiphi and Typhlodromus pyri) should be tested. In addition to these, two additional species must also be tested, which should be relevant to the intended use of the substance. Where possible and if appropriate, they should represent the other two major functional groups, ground dwelling predators and foliage dwelling predators. Where effects are observed with species relevant to the proposed use of the product, further testing may be carried out at the extended laboratory/semi-field level. Selection of the relevant test species should follow the proposals outlined in Setac — Guidance document on regulatory testing procedures for pesticides with non-target arthropods (12) . Testing must be conducted at rates equivalent to the highest rate of field application to be recommended.
Where relevant, testing should be done according to appropriate guidelines which satisfy at least the requirements for testing as included in Setac — Guidance document on regulatory testing procedures for pesticides with non-target arthropods.
The test should provide the LC 50 value of the active substance to earthworms, where possible the highest concentration causing no mortality and the lowest concentration causing 100 % mortality, and must include observed morphological and behavioural effects.
Effects on earthworms must be investigated, where preparations containing the active substance are applied to soil, or can contaminate soil.
The test must be carried out in accordance with Commission Directive 88/302/EEC (13) adapting to technical progress for the ninth time Council Directive 67/548/EEC on the approximation of laws, regulations and administrative provisions relating to the classification, packaging and labelling of dangerous substances, Part C, Toxicity for earthworms: Artificial soil test.
The test should provide the NOEC and the effects on growth, reproduction and behaviour.
Where on the basis of the proposed manner of use of preparations containing the active substance or on the basis of its fate and behaviour in soil (DT 90 > 100 days), continued or repeated exposure of earthworms to the active substance, or to significant quantities of metabolites, degradation or reaction products, can be anticipated expert judgement is required to decide whether a sublethal test can be useful.
The test must be carried out on Eisenia foetida.
The test should provide sufficient data to evaluate the impact of the active substance on soil microbial activity, in terms of nitrogen transformation and carbon mineralization.
The test must be carried out where preparations containing the active substance are applied to soil or can contaminate soil under practical conditions of use. In the case of active substances intended for use in preparations for soil sterilization, the studies must be designed to measure rates of recovery following treatment.
Soils used must be freshly sampled agricultural soils. The sites from which soil is taken must not have been treated during the previous two years with any substance that could substantially alter the diversity and levels of microbial populations present, other than in a transitory manner.
Setac — Procedures for assessing the environmental fate and ecotoxicity of pesticides.
A summary of available data from preliminary tests used to assess the biological activity and dose range finding, whether positive or negative, which may provide information with respect to possible impact on other non-target species, both flora and fauna, must be provided, together with a critical assessment as to its relevance to potential impact on non-target species.
Effects on biological methods for sewage treatment must be reported where the use of plant protection products containing the active substance can give rise to adverse effects on sewage treatment plants.]
Textual Amendments
Hazard symbol(s)
Indications of danger
Risk phrases
Safety phrases
Substance within the meaning of the definition of Article 2, point 3.
Standardized System of Nomenclature and Diagnostic Criteria — Guides for Toxicologic Pathology
Guidelines for Good Epidemiology Practices for Occupational and Environmental Research, developed by the Chemical Manufacturers Association's Epidemiology Task Group, as part of the Epidemiology Resource and Information Centre (ERIC), Pilot Project, 1991]
[F4Guidance under development.]
[F5Society of Environmental Toxicology and Chemistry (SETAC), 1995. Procedures for assessing the environmental fate and ecotoxicity of pesticides, ISBN 90-5607-002-9.]
[F6Society of Environmental Toxicology and Chemistry (Setac), 1995. Procedures for Assessing the Environmental Fate and Ecotoxicity of Pesticides, ISBN 90-5607-002-9.
From the Workshop European Standard Characteristics of beneficials Regulatory Testing (Escort), 28 to 30 March 1994 , ISBN 0-95-22535-2-6.
Textual Amendments
F1 Substituted by Commission Directive 94/37/EC of 22 July 1994 amending Council Directive 91/414/EEC concerning the placing of plant protection products on the market.
F3 Substituted by Commission Directive 94/79/EC of 21 December 1994 amending Council Directive 91/414/EEC concerning the placing of plant protection products on the market.
F4 Substituted by Commission Directive 96/68/EC of 21 October 1996 amending Council Directive 91/414/EEC concerning the placing of plant protection products on the market (Text with EEA relevance).