Animal testing versus calculation method
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1 Animal testing versus calculation method Dr. Tamara Coja, MSc Toxicology Institute for Plant Protection Products, Department for Toxicology Interzonal workshop Harmonisation of risk assessment in section toxicology Vienna, June 2015 Austrian Agency for Health and Food Safety
2 Outline European Legislations and animal testing Data requirements for testing of PPPs according to Regulation (EU) No 284/2013 Examples: I ) Necessity or no-necessity of acute inhalation toxicity study II) Vertebrate study versus calculation method 2
3 European Legislations and animal testing 3
4 Animal testing Directive 2010/63/EU (Legislation for the protection of animals used for scientific purposes), Article 47: 1. The Commission and the Member States shall contribute to the development and validation of alternative approaches which could provide the same or higher levels of information as those obtained in procedures using animals, but which do not involve the use of animals or use fewer animals (). 4. Member States shall, at national level, ensure the promotion of alternative approaches and the dissemination of information thereon. 4
5 REACH Data sharing Companies must share information on their hazardous properties and jointly submit this hazard information to ECHA Alternative methods and approaches Companies can use existing animal studies, conducted before REACH. They can also predict the properties of substances by read-across approach 5
6 REACH ECHA testing proposals: QSAR Toolbox: 6
7 Regulation (EC) No 1223/2009 The Cosmetics Regulation prohibits the placing on the market of cosmetic products, or products containing ingredients, which have been tested on animals to meet the requirements of that regulation using a method other than a validated alternative method 7
8 Regulation (EC) No 1272/2008 (27) The classification and labelling criteria set out in this Regulation should take the utmost account of promoting alternative methods for the assessment of hazards of substances and mixtures and of the obligation to generate information on intrinsic properties by means other than tests on animals within the meaning of Directive 86/609/EEC (now: Directive 2010/63/EU) as laid down in Regulation (EC) No 1907/
9 Article 62 of Regulation (EC) No 1107/2009 Article 62 (Sharing of tests and studies involving vertebrate animals): 1. Testing on vertebrate animals for the purposes of this Regulation shall be undertaken only where no other methods are available. Duplication of tests and studies on vertebrates undertaken for the purposes of this Regulation shall be avoided in accordance with paragraphs 2 to Member States shall not accept duplication of tests and studies on vertebrate animals or those initiated where conventional methods (Remark: calculation method is meant) described in Annex II to Directive 1999/45/EC could reasonably have been used, in support of applications for authorisations. 9
10 Data requirements PPP Regulation (EU) No 284/
11 Data requirements PPP - Regulation (EU) No 284/ Tests on vertebrate animals shall be undertaken only where no other validated methods are available. Alternative methods: in vitro methods and in silico methods; reduction and refinement methods for in vivo testing (3Rs) 11
12 Data requirements PPP - Regulation (EU) No 284/2013 Acute oral toxicity A test for acute oral toxicity shall be carried out, unless the applicant can justify an alternative approach under Regulation (EC) No 1272/2008 Acute dermal toxicity A test for dermal toxicity shall be carried out on a case by case basis, unless the applicant can justify an alternative approach under Regulation (EC) No 1272/
13 Data requirements PPP - Regulation (EU) No 284/2013 Acute inhalation toxicity The study shall provide the inhalation toxicity to rats of the plant protection product or of the smoke it generates The study shall be carried out where the plant protection product: (a) is a gas or liquified gas; (b) is a smoke generating plant protection product or fumigant; (c) is used with fogging/misting equipment; (d) is a vapour releasing plant protection product; (e) is supplied in an aerosol dispenser; 13
14 Data requirements PPP - Regulation (EU) No 284/2013 (f) is in a form of a powder or granules containing a significant proportion of particles of diameter < 50 µm (> 1 % on a weight basis); (g) is to be applied from aircraft in cases where inhalation exposure is relevant; (h) contains an active substance with a vapour pressure > Pa and is to be used in enclosed spaces such as warehouses or glasshouses; (i) is to be applied by spraying. A study shall not be required if the applicant can justify an alternative approach under Regulation (EC) No 1272/2008, where applicable 14
15 Data requirements PPP - Regulation (EU) No 284/2013 Skin irritation The skin irritancy of the plant protection product shall be reported based on the tiered approach, unless the applicant can justify an alternative approach under Regulation (EC) No 1272/2008. First: weight of evidence if study necessary Tiered approach: (1) the assessment of dermal corrosivity using a validated in vitro test method; (2) the assessment of dermal irritation using a validated in vitro test method (such as human reconstituted skin models); (3) an initial in vivo dermal irritation study using one animal, and where no adverse effects are noted; (4) confirmatory testing using one or two additional animals. 15
16 Data requirements PPP - Regulation (EU) No 284/2013 Eye irritation Eye irritation tests shall be provided, unless it is likely that severe effects on the eyes may be produced or the applicant can justify an alternative approach under Regulation (EC) No 1272/2008. First: weight of evidence if study necessary Tiered approach: (1) the use of an in vitro dermal irritation/corrosion test to predict eye irritation/corrosion; (2) the performance of a validated or accepted in vitro eye irritation study to identify severe eye irritants/ corrosives (such as BCOP, ICE, IRE, HET-CAM), and where negative results are obtained; 16
17 Data requirements PPP - Regulation (EU) No 284/2013 Eye irritation (3) the assessment of eye irritation using an available, in vitro test method validated for plant protection products for identification of nonirritants or irritants, and when not available; (4) an initial in vivo eye irritation study using one animal, and where no adverse effects are noted; (5) confirmatory testing using one or two additional animals. 17
18 Data requirements PPP - Regulation (EU) No 284/2013 Skin sensitisation The skin sensitisation test shall be carried out unless the active substances or co-formulants are known to have sensitising properties or the applicant can justify an alternative approach under Regulation (EC) No 1272/
19 Data requirements PPP - Regulation (EU) No 284/2013 Prerequisite for applicability of calculation method for estimation of acute toxicity, irritation and sensitisation: toxicological properties of all components shall be provided or reliably predicted by a validated method 19
20 Available OECD Guidelines in vitro Skin corrosion/irritation (latest updates): 26 Sep 2014: Test No. 431: In Vitro Skin Corrosion: Reconstructed Human Epidermis (Rhe) Test Method 26 July 2013: Test No. 430: In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test Method (TER) 26 July 2013: Test No. 431: In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method 26 July 2013: Test No. 439: In Vitro Skin Irritation - Reconstructed Human Epidermis Test Method 20
21 Available OECD Guidelines in vitro Eye damage/irritation (latest updates): 26 July 2013: Test No. 437: Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage 26 July 2013: Test No. 438: Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage 02 Oct 2012: Test No. 460: Fluorescein Leakage Test Method for Identifying Ocular Corrosives and Severe Irritants 21
22 Available OECD Guidelines in vitro Skin sensitisation (new): 05 Feb 2015:Test No. 442C: In Chemico Skin Sensitisation 05 Feb 2015:Test No. 442D: In Vitro Skin Sensitisation 22
23 Regulatory real life examples Acute inhalation toxicity 23
24 Acute inhalation toxicity What is meant in Regulation (EC) 284/2013 by: acute inhalation toxicity study is necessary if PPP is to be applied by spraying? ( Old Commission Regulation (EU) No 545/2011: if PPP is to be applied in a manner which generates a significant proportion of particles or droplets of diameter < 50 µm (> 1 % on a weight basis) ). Particle size in the acute inhalation toxicity study 1 4 µm Particle size for spraying in e.g grapes (fine particles) recommended at 350 µm and should not be < 125 µm Particles > 100 µm have high settling velocity, so exposure unlikely to occur 24
25 Acute inhalation toxicity 1272/2008: The ECHA guidance document on the application of CLP criteria (pages , June 2015) states: () The use of highly respirable dusts and mists is ideal to fully investigate the potential inhalation hazard of the substance. However, it is acknowledged that these exposures may not necessarily reflect realistic conditions. For instance, solid materials are often micronised to a highly respirable form for testing, but in practice exposures will be to a dust of much lower respirability.() In such situations, specific problems may arise with respect to classification and labelling, as these substances are tested in a form (i.e. specific particle size distribution) that is different from all the forms in which these substances are placed on the market and in which they can reasonably be expected to be used (). 25
26 Acute inhalation toxicity So, trigger for inhalation testing is exposure-related A philosophical question: if no acute inhalation study is necessary according to data requirements and no exposure expected, than we are not allowed to classify based on properties of active or coformulants, right? 26
27 Acute inhalation toxicity C&L of active XY T, R23; Toxic by inhalation Acute Tox 2, H330; Fatal if inhaled Formulatio n type Content of active XY (%) in PPP WG 70 Grapes, spraying GAP Remark % active XY in spray dilution PPP nearly dust free, no attrition < 0.1% particles of respirable size (< 10 µm) < 0.1% particles < 50 µm 0.05% to 0.375% zrms decided not to classify for acute inhalation toxicity and received mostly disagreements by cms 27
28 Regulatory real life examples Unnecessary vertebrate studies versus calculation method 28
29 Vertebrate studies versus calculation method Component in PPP C&L Content (%) in PPP Formulation type GAP Remarks Active substance Formulant 1 Formulant 2 Xn, R22/20 H302 H332 R41 H318 Well-known food additive 15 SG, nearly dust free Field crops (cereals, etc..) 82 LD50 oral > 5000 No skin irritant No skin sensitiser No data on acute dermal and acute inhalation toxicity
30 Vertebrate studies versus calculation method zrms accepted all six submitted vertebrate studies Calculation method would lead us solely to: R41 / H318 Studies showed: R41 / H318 and R22 / H302 What would we miss if we loose R22 / H302? 1) P270: Do not eat, drink or smoke when using the product (GAP anyhow?) optional for professional users according to Guidance on Labelling and Packing in accordance with Regulation (EC) No 1272/2008 2) P264 Wash thoroughly after handling (if hands are meant, GAP anyhow?) - optional 3) P301 + P312 IF SWALLOWED: Call a POISON CENTER or doctor/physician if you feel unwell - optional 4) P330 Rinse mouth - optional 30
31 Food for thoughts? Acute oral toxicity study calculation method in most cases easily applicable Acute dermal toxicity study waiving easily applicable (results of acute oral study and skin irritation) Acute inhalation toxicity study do we need it if there is no realistic exposure? Skin and eye irritation study calculation method applicable case by case, certainly applicable when co-formulants classified Skin sensitisation probably most difficult to predict, unless information on known sensitiser available 31
32 Food for thoughts? How much information do we loose if we mistakenly not classify for acute toxicity, irritation and sensitisation? What about risk mitigation measures in uncertain cases (e.g. wear eye protection if calculation method uncertain or not sufficient data on co-formulants) instead of vertebrate study? Appeal to the Member States and the industry: let us think twice before we ask for /conduct an unnecessary animal study 32
33 To start the expert discussions CIRCABC expert forum established for exchange of knowledge, questions, doubts (Member States) CIRCABC PPP Zonal Newsgroups Toxicology and Operator & Bystander Safety Let`s start live it! 33
Ian Indans Regulatory scientist Chemicals Regulation Directorate (CRD)
Health and Safety Executive Ian Indans Regulatory scientist Chemicals Regulation Directorate (CRD) Health and Safety Executive Acute Inhalation Toxicity testing; The 3Rs, Current needs and Future Prospects
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