permit a decision to be made as to whether, or not, the active substance can be approved,
specify appropriate conditions or restrictions to be associated with any approval,
classify the active substance as to hazard,
establish a relevant acceptable daily intake (ADI) level for man,
establish acceptable operator exposure level(s) (AOEL),
specify the pictograms, signal words, and relevant hazard and precautionary statements for the protection of man, animals and the environment to be included in packaging (containers),
identify relevant first aid measures as well as appropriate diagnostic and therapeutic measures to be followed in the event of poisoning in man, and
permit an evaluation to be made as to the nature and extent of the risks for man, animals (species normally fed and kept or consumed by man) and of the risks for other non-target vertebrate species.
Quite limited data, as described below and restricted to one test species (normally the rat) may be all that is required in this area. These data can provide information useful in the design and interpretation of subsequent toxicity tests. However, it must be remembered that information on interspecies differences may be crucial in extrapolation of animal data to man and information on percutaneous penetration, absorption, distribution, excretion and metabolism may be useful in operator risk assessments. It is not possible to specify detailed data requirements in all areas, since the exact requirements will be dependent upon the results obtained for each particular test substance.
Aim of the test:
The tests shall provide sufficient data to permit:
an evaluation of the rate and extent of absorption,
an evaluation of the tissue distribution and the rate and extent of excretion of the test substance and the relevant metabolites,
the identification of metabolites and the metabolic pathway.
The effect of dose level on these parameters and whether the results are different after single versus repeated doses, shall also be investigated.
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).
Regulation (EC) No 440/2008, Method B 36, 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 Regulation (EC) No 1272/2008. The information generated through acute toxicity testing is of particular value in assessing hazards likely to arise in accident situations.
The acute oral toxicity of the active substance must always be reported.
The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 1 bis or B 1 ter.
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 the Annex to Regulation (EC) No 440/2008, method B 3.
The inhalation toxicity of the active substance must be reported where the active substance is:
a gas or liquified gas,
is to be used as a fumigant,
is to be included in a smoke generating, aerosol or vapour releasing preparation,
is to be used with fogging equipment,
has a vapour pressure > 1 × 10–2 Pa and is to be included in preparations to be used in enclosed spaces such as warehouses or glasshouses,
is to be included in preparations which are powders containing a significant proportion of particles of diameter < 50 μm (> 1 % on a weight basis), or
is to be included in preparations to be applied in a manner which generates a significant proportion of particles or droplets of diameter < 50 μm (> 1 % on a weight basis).
The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 2.
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 the Annex to Regulation (EC) No 440/2008, Method B 4.
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 the Annex to Regulation (EC) No 440/2008, Method B 5.
The test will provide sufficient information to assess the potential of the active substance to provoke skin sensitisation reactions.
The test must always be carried out except where the substance is a known sensitiser.
The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 6.
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 the Annex to Regulation (EC) No 440/2008, Method B 7.
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.
The test must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Methods B 26 and B 27, sub-chronic oral toxicity test repeated dose 90.
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: the Annex to Regulation (EC) No 440/2008, Method B 9, repeated dose toxicity (dermal),
90-day dermal: the Annex to Regulation (EC) No 440/2008 Method B 28, sub-chronic dermal toxicity study,
28-day inhalation: the Annex to Regulation (EC) No 440/2008, Method B 8, repeated dose toxicity (inhalation),
90-day inhalation: the Annex to Regulation (EC) No 440/2008, Method B 29, sub-chronique inhalation toxicity study.
These studies are of value in:
the prediction of genotoxic potential
the early identification of genotoxic carcinogens
the elucidation of the mechanism of action of some carcinogens.
To avoid responses that are artefacts of the test system, excessively toxic doses must not be used in either in vitro or in vivo assays for mutagenicity. This approach shall be regarded as general guidance. It is important that a flexible approach is adopted, with selection of further tests which are dependent upon interpretation of results at each stage.
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:
Annex to Regulation (EC) No 440/2008, Method B 13/14 — reverse mutation test using bacteria,
Annex to Regulation (EC) No 440/2008, Method B 10 — in vitro mammalian chromosome aberration test,
Annex to Regulation (EC) No 440/2008, Method B 17 — in vitro mammalian cell gene mutation test.
If all the results of the in vitro studies are negative further testing must be done with consideration of other relevant information available (including toxicokinetic, toxicodynamic and physico-chemical data and data on analogous substances). The test can be an in vivo study or an in vitro study using a different metabolising system from that/those previously used.
If the in vitro cytogenetic test is positive, an in vivo test using somatic cells (metaphase analysis in rodent bone marrow or micronucleus test in rodents) must be conducted.
If either of the in vitro gene mutation tests are positive, an in vivo test to investigate unscheduled DNA synthesis or a mouse spot test must be conducted.
The following test guidelines are acceptable:
Annex to Regulation (EC) No 440/2008, Method B 12 — In vivo mammalian erythrocyte micronucleus test,
Annex to Regulation (EC) No 440/2008, Method B 24— Mouse spot test,
Annex to Regulation (EC) No 440/2008, Method B 11 — In vivo Mammalian Bone-Marrow chromosome aberration test.
When any result of an in vivo study in somatic cells is positive, in vivo testing for germ cell effects may be justified. The necessity for conducting these tests will have to be considered on a case by case basis, taking into account information regarding toxicokinetics, use and anticipated exposure. Suitable tests would need to examine interaction with DNA (such as the dominant lethal assay), to look at the potential for inherited effects and possibly make a quantitative assessment of heritable effects. It is recognised that in view of their complexity, the use of quantitative studies would require strong justification.
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 (2 years) of the active substance must be conducted using the rat as test species; these studies can be combined.
A carcinogenicity study of the active substance must be conducted using the mouse as test species.
Where a non-genotoxic mechanism for carcinogenicity is suggested, a well argued case, supported with relevant experimental data, including that necessary to elucidate the possible mechanism involved, must be provided.
While the standard reference points for treatment responses are concurrent control data, historical control data, may be helpful in the interpretation of particular carcinogenicity studies. Where submitted, historical control data shall be from the same species and strain, maintained under similar conditions and shall be from contemporaneous studies. The information on historical control data provided must include:
identification of species and strain, name of the supplier, and specific colony identification, if the supplier has more than one geographical location,
name of the laboratory and the dates when the study was performed,
description of the general conditions under which animals were maintained, including the type or brand of diet and, where possible, the amount consumed,
approximate age, in days, of the control animals at the beginning of the study and at the time of killing or death,
description of the control group mortality pattern observed during or at the end of the study, and other pertinent observations (e.g. diseases, infections),
name of the laboratory and the examining scientists responsible for gathering and interpreting the pathological data from the study, and
a statement of the nature of the tumours that may have been combined to produce any of the incidence data.
The doses tested, including the highest dose tested, must be selected on the basis of the results of short-term testing and where available at the time of planning the studies concerned, on the basis of metabolism and toxicokinetic data. The highest dose level in the carcinogenicity study shall elicit signs of minimal toxicity such as slight depression in body-weight gain (less than 10 %), without causing tissue necrosis or metabolic saturation and without substantially altering normal lifespan due to effects other than tumours. If the long-term toxicity study is carried out separately, the highest dose level shall elicit definite signs of toxicity without causing excessive lethality. Higher doses, causing excessive toxicity are not considered relevant to evaluations to be made.
In the collection of data and compilation of reports, incidence of benign and malignant tumours must not be combined, unless there is clear evidence of benign tumours becoming malignant with time. Similarly, dissimilar, un-associated tumours, whether benign or malignant, occurring in the same organ, must not be combined, for reporting purposes. In the interests of avoiding confusion, terminology such as that developed by the American Society of Toxicologic Pathologists, or the Hannover Tumour Registry (RENI) shall be used in the nomenclature and reporting of tumours. The system used must be identified.
It is essential that biological material selected for histopathological examination includes material selected to provide further information on lesions identified during gross pathological examination. Where relevant to the elucidation of mechanism of action and available, special histological (staining) techniques, histochemical techniques and electron microscopic examinations, must be conducted and reported.
The studies must be carried out in accordance with the Annex to Regulation (EC) No 440/2008, Method B 30 Chronic toxicity test, Method B 32 Carcinogenicity test or Method B 33 combined chronic toxicity/carcinogenicity test.
The adverse reproductive effects are of two main types:
impairment of male or female fertility, and
impacts on the normal development of progeny (developmental toxicity).
Possible effects on all aspects of reproductive physiology in both males and females, as well as possible effects on pre-natal and post-natal development, must be investigated and reported. If in exceptional circumstances, it is claimed that such testing is unnecessary, that claim must be fully justified.
While the standard reference point for treatment responses are concurrent control data, historical control data may be helpful in the interpretation of particular reproductive studies. Where submitted, historical control data shall be from the same species and strain, maintained under similar conditions and shall be from contemporaneous studies. The information on historical control data provided must include:
identification of species and strain, name of the supplier, and specific colony identification, if the supplier has more than one geographical location,
name of the laboratory and the dates when the study was performed,
description of the general conditions under which animals were maintained, including the type or brand of diet and, where possible, the amount consumed,
approximate age, in days, of the control animals at the beginning of the study and at the time of killing or death,
description of the control group mortality pattern observed during or at the end of the study, and other pertinent observations (e.g. diseases, infections), and
name of the laboratory and the examining scientist responsible for gathering and interpreting the toxicological data from the study.
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 the Annex to Regulation (EC) No 440/2008, Method B 35, two-generation reproduction toxicity study. In addition organ weight of reproductive organs must be reported.
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 shall 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 the Annex to Regulation (EC) No 440/2008, Method B 31, prenatal developmental toxicity study.
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 [F2the EU-derived domestic legislation which transposed] Article 10 of Council Directive 98/24/EC, practical data and information relevant to the recognition of the symptoms of poisoning, and on the effectiveness of first aid and therapeutic measures have to be submitted. More specific references to the investigation for antidotal pharmacology or safety pharmacology using animals shall be provided. Where relevant, the effectiveness of potential antagonists to poisoning, shall be investigated and reported.
Textual Amendments
F2Words in Annex Pt. A point 5.9 inserted (31.12.2020) by The Plant Protection Products (Miscellaneous Amendments) (EU Exit) Regulations 2019 (S.I. 2019/556), regs. 1(1), 16(5)(b)(vi); 2020 c. 1, Sch. 5 para. 1(1)
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 shall, where feasible, include data relevant to the mechanism of action of the active substance. These reports shall, where available, include data from persons exposed in manufacturing plants or after application of the active substance (e.g. in efficacy trials).
Available information on the sensitisation including allergenic response of workers and others exposed to the active substance, must be provided, and include where relevant details of any incidence of hypersensitivity. The information provided shall include details of frequency, level and duration of exposure, symptoms observed and other relevant clinical information.
Available reports from the open literature, relating to clinical cases and poisoning incidents, where they are from refereed journals or official reports, must be submitted together with reports of any follow-up studies undertaken. Such reports shall contain complete descriptions of the nature, level and duration of exposure, as well as the clinical symptoms observed, first aid and therapeutic measures applied and measurements and observations made. Summary and abstract information is not of value.
Where supported with the necessary level of detail, such documentation can be of particular value in confirming the validity of extrapolations from animal data to man and in identifying unexpected adverse effects which are specific to humans.
Where available, and supported with data on levels and duration of exposure, and conducted in accordance with recognised standards, 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 (point 1.11) and any bridging studies conducted (point (iv) of the introduction to Section 5), the relevance of the data as submitted to the assessment of the toxicological profile of the active substance as manufactured, must be argued.
On the basis of an assessment of the data base, and the relevant decision making criteria and guidelines, justifications must be submitted for the Noaels proposed for each relevant study.
On the basis of these data scientifically reasoned proposals for the establishment of ADI and AOEL(s) for the active substance must be submitted.]
Textual Amendments applied to the whole legislation
F1Repealed (subject to transitional measures in Arts. 3, 4) by Commission Regulation (EU) No 283/2013 of 1 March 2013 setting out the data requirements for active substances, in accordance with Regulation (EC) No 1107/2009 of the European Parliament and of the Council concerning the placing of plant protection products on the market Text with EEA relevance.