GERMAN FEDERAL INSTITUTE

FOR RISK ASSESSMENT

The toxicological evaluation of pesticides

Data requirements in the EU and principles of

risk assessment for consumer protection and
occupational safety

Dr Glenn Lurman
Toxicology of Active Substances and their Metabolites
Department Safety of Pesticides
German Federal Institute for Risk Assessment, Berlin
What (I hope) you will take away from this…

Which data are needed for hazard (and risk) charaterisation

Relevant toxicological end-points

An understanding of what references values are AND how they

are derived

Page 2
What are we talking about? Pesticides

Active substances in plant protection products,

mainly intended for agricultural use (in contrast
to „biocides“ that are applied in other fields
such as desinfection, wood protection,
household insect control…)

Main groups: Fungicides, Herbicides,

Insecticides, Acaricides, Growth regulators,
Molluscicides, Nematicides, (Rodenticides)

Page 3
Benefits and harm of plant protection products
On the one hand…
• Higher crop yields
• Reduced stored food and feed
losses
• Consumer health protection
(e.g., prevention of fungal
diseases or vector-transmitted
infections)
• Better quality products
But, on the other …
• Exposure of operators,
bystanders, residents
• (Potential) health effects of
residues on consumers
• Ecological damage (toxic to
vertebrates and invertebrates
and plants)
• Water and soil pollution
• Reduced biodiversity
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Risk assessment
Define the problem

Hazard
Use
Identification

Hazard
Exposure
Characterisation

Risk
Characterisation

Communication

Page 5
Who to protect?
Define the problem

Risk
Characterisation

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 Illustration created with http://www.tagxedo.com
Who needs to be protected?

Foetuses
Toddlers
Children
Teenagers
Adults (men/women)
Elderly
Sick
Disabled
Pregnant women
People with specific vulnerabilities

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Protect against what?
Define the problem

Exposure route and duration

Toxic effects

Risk
Characterisation

Page 8
Focus on human health: Hazard – Risk – Exposure (I)

Illustration created with http://www.tagxedo.com

What are the toxic properties of a pesticide?

Acute toxicity
Skin and eye irritation / corrosion
Allergy
Mutation
Cancer
Malformations
Retardations
Fertility
Effect on lactation
Neurotoxicity
Organ damage
…

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Focus on human health: Hazard – Risk – Exposure (II)

Exposure:
To what extent will humans be exposed?

Who? Consumers, operators, bystanders,

workers, or residents
… by which route? Oral and/or dermal and/or
inhalation?
… in what amount? (And how much will become
systemically available?)
… how often?

Cave: Aggregate and cumulative exposures

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Focus on human health: Hazard – Risk – Exposure (III)

R
I
Toxicity S Exposure
K

Principle of risk assessment:

Exposure is compared to “safe doses“,
i.e., the toxicological reference values

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The scope of our toxicological considerations …

1. The active substance(s) = a.s. in a plant

protection product (PPP), also called the
“active (ingredient)“, “compound“, or
“pesticide“, typically has an extensive
toxicological database.
2. PPP may contain more than one a.s. and
typically co-formulants (e.g. solvents,
surfactants, synergists). Much less data
for co-formulants.
3. Impurities with few data, if any.
4. Metabolites (in plants, animals, raw food,
processed food, soil, groundwater) with a
variable database.

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Toxicological studies with an active ingredient

Data requirements in the EU: Regulation

(EU) No 283/2013 (active substances)
and No 284/2013 (products)
Legal basis in the EU: Regulation (EC) No
1107/2009

Toxicological studies shall be sufficient …

• to derive reference values as basis for risk
assessment
• to determine a possible need for
classification and labelling
• to check if “cut-off“ criteria may apply

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 Toxicological studies (in general)

… shall be performed according to

OECD test guidelines
(or other acceptable methods) and
under GLP conditions
if conducted by the applicant

Internationally defined and recognised

method for assessing substances for
given hazards.

Methods are (usually) validated,

transferable, robust and of sufficient
statistical power.

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Data requirements for active ingredients

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Evaluation of an active substance: endpoints to be covered (I)

•Toxicokinetics and metabolism (Absorption,

Distribution, Metabolism and Excretion = ADME),
including comparative in vitro studies, in future also
kinetic data from feeding studies

•Acute toxicity (Effects after single administration

via different routes)

•Skin and eye irritation

•Skin sensitisation (allergenicity)

•[Phototoxicity (in vitro, depending on optical absorption

properties and distribution of the substance to the eyes
and certain skin areas)]

„Tox 6-Pack“

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Evaluation of an active substance: endpoints to be covered (II)

•Short-term toxicity (studies of 28 or 90 days via

the oral and possibly also other routes) in rodents
and non-rodents (mainly dogs)

•Genotoxicity (in vitro in different test systems

and in vivo)

•Long-term (chronic) toxicity (mostly in rats)

•Carcinogenicity (in principle in two species,

usually in rats and mice)

•Reproductive toxicity (effects on fertility,

reproductive performance, offspring development;
parental toxicity, usually in rats)

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Evaluation of an active substance: endpoints to be covered (III)

•Developmental toxicity (including teratogenicity,

two species, usually rats and rabbits)

•Neurotoxicity and delayed neuropathy

•Mechanistic studies (optional, depending on

effects in routine studies) including those for
elucidation and characterization of endocrine
disrupting properties

•Medical data (Observations in humans from

different sources)

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Toxicological data requirements for products

•Acute toxicity (oral, dermal and

inhalation) Animal experiments
or calculations based
•Skin and eye irritation on results on the
components of the
•Skin sensitisation mixture

•Dermal absorption

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 Toxicity of metabolites

•Needed if the metabolites of concern (in plants,

food, groundwater …) do not occur in laboratory
animals or only in traces

•Requirements depend on whether metabolites

occur in residues (of edible commodities) or in
groundwater§

•Genotoxicity studies are a usual requirement

•Further studies (acute and short-term toxicity
developmental toxicity, carcinogenicity, …) may be
required if necessary

§, EFSA Panel on Plant Protection Products and their Residues (PPR); Scientific Opinion on Evaluation of the Toxicological Relevance of Pesticide
Metabolites for Dietary Risk Assessment. EFSA Journal 2012;10(07): 2799.
EFSA Panel on Plant Protection Products and their Residues (PPR); Guidance on the establishment of the residue definition for dietary risk
assessment. EFSA Journal 2016;14(12):4549, 129 pp. doi:10.2903/j.efsa.2016.4549
European Commission: Guidance document on the assessment of the relevance of metabolites in groundwater of substances regulated under Council
Directive 91/414/EEC; Sanco/221/2000 –rev.10- final; 25 February 2003
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NOAEL / LOAEL

NOAEL (No Observed Adverse Effect Level)

An exposure level at which there are no statistically or biologically significant increases in the frequency or
severity of adverse effects above an appropriate control; some effects may be produced at this level, but they
are not considered as adverse or precursors to adverse effects.

LOAEL (Lowest Observed Adverse Effect Level):

The lowest dose or exposure level of a chemical in a study at which there is a statistically or biologically
significant increase in the frequency or severity of an adverse effect in the exposed population as compared
with an appropriate, unexposed control group

(http://www.epa.gov/ncea/bmds/bmds_training/appendices/glossary.htm)
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Benchmark Dose (BMD)

BMD
Observed mean responses are plotted with confidence intervals or SD. A dose-response model is fitted,
together with 95% confidence interval. The benchmark response (BMR) corresponds to the point at which a
“measureable change” occurs, e.g. 5% for continuous data, 10% for discrete data, or effect-specific biological
relevance. BMDL (and BMDU) values are derived from the lower (and upper) confidence values respectively.

(EFSA Update: Use of the Benchmark Dose Approach in Risk Assessment, EFSA Journal 2017;15(1):4658)

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Benchmark Dose (BMD)

BMDU
<3
BMDL

BMDU > 3
BMDL

BMD
Observed mean responses are plotted with confidence intervals or SD. A dose-response model is fitted,
together with 95% confidence interval. The benchmark response (BMR) corresponds to the point at which a
“measureable change” occurs, e.g. 5% for continuous data, 10% for discrete data, or effect-specific biological
relevance. BMDL (and BMDU) values are derived from the lower (and upper) confidence values respectively.

(EFSA Update: Use of the Benchmark Dose Approach in Risk Assessment, EFSA Journal 2017;15(1):4658)

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Information to be obtained (I)

What are the target organs for toxic effects?

Which toxic (adverse) effects occur? (Are these

effects relevant to humans?)

At which doses do they occur?

Is there a dose-response relationship?

What is the “lowest observable adverse effect

level“ (LOAEL)?

What is the “no-observed adverse effect level“

(NOAEL); i.e., the highest dose without any
adverse effect?

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Information to be obtained (II)

When are adverse effects observed for the first time?

Do they occur after a single exposure or after a
longer period?

Are there species differences? (Most suitable

model?)

Are there sex differences?

Are pregnant or young animals more sensitive?

Does the route of administration make a difference?

Are the effects reversible?

Mechanism or mode of action?

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Identification of effects – General findings

•Mortality

•Clinical signs

•Altered behaviour

•Body weight (gain) changes (mostly ↓)

•Alterations in food consumption or food efficiency

•Haematological findings (such as anaemia)

•Alterations in clinical-chemistry parameters

(blood and urine)

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Identification of effects – Post mortem findings

•Macroscopic changes at necropsy

•Organ weight changes (absolute / relative)

•Histopathological lesions

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Identification of effects – Endpoint-specific

•Positive for genotoxicity in vitro/in vivo

•More frequent chromosome aberrations in vitro/in
vivo
•Specific neurotoxic potential
(neurohistopathological lesions, behavioural
changes, performance in neurological tests ↓)
•Lower fertility
•Impaired reproductive success
•Offspring mortality
•Lower pup weight (gain)
•Lower fetal weight
•Malformations or more frequent skeletal or
visceral variations

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Are these effects toxicologically relevant?

•Severity of the effect (Severity of the lesions?

How many animals in the dose group are
affected? What will be the impact on health and
reproduction?)

•Is the difference statistically significant compared

to the control group?

•Is there a dose-response relationship?

•Is the effect biologically plausible?

•Relevant for humans?

•Is the control incidence within the range of the

historical control data (and the treated group
incidence outside)?

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Sources of information

1. Original studies that have to be

conducted according to GLP and
compliant with OECD Test Guidelines (The
“core“ or “apical“ studies as a minimum)

2. Summaries and evaluations of tox studies

by supranational organisations like WHO
and FAO (JMPR Reports), well-
recognised agencies such as U.S. EPA
(“Pesticide Fact Sheets“) or EFSA
(“Conclusions“), etc

3. Published literature of which the

reliability depends on the level of detail in
reporting and if it was clearly described
what has been done

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Reference values

Who to protect?
adults, pregnant women, children, …
What exposure route?
dietary, non-dietary (oral, dermal, inhalation)
Which exposure duration?
acute, medium-term, long-term
Other factors to consider?
systemic or external RfD

Select relevant point of departure (PoD)

Select relevant uncertainty factor (UF)
Corrrect for limited bioavailability, where relevant

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Reference values, ADI

Acceptable Daily Intake, given in mg kg body

weight (bw) per day (d), i.e., the dose to which a
person may be exposed to daily, from all possible
sources, during a lifetime without any impact on health.

•Needed for all pesticides (active substances) for which

dietary exposure cannot be excluded.
•Generally based on NOAELs in long-term studies.
•For substances of high acute toxicity or specific repro-
toxic potential, other studies may be considered.
•Usual uncertainty factor of 100 (×10 inter-species
variability, ×10 intra-species variability) may be
increased in the case of severe effects, data gaps, or if
no NOAEL could be established and the LOAEL must
be used as the PoD.
PoD
ADI =
Currently no agreed EU guidance document available
UF
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Reference values, ARfD

Acute Reference Dose, given in mg kg bw-1, is the

amount of a substance that may be ingested in 24 hours
without any adverse effects

•Not needed for all active ingredients, only if there is

evidence of adverse effects after a single or few
administrations
•The PoD usually comes from effects seen in acute,
short-term, neurotoxicity or developmental studies
•May be much higher than the ADI (but sometimes is
not)
•Mostly based on NOAELs in short-term or
developmental studies or in the acute neurotoxicity
study; acute oral toxicity study not appropriate (=>
LD/LC50)

PoD
ARfD =
UF
Currently no agreed EU guidance document available
Page 33
Reference values, AOEL

Acceptable Operator Exposure Level, given in mg

kg bw-1 d-1; is the reference value for occupational
safety that has to be compared to the exposure of
operators, workers, bystanders and residents

•Needed for all active substances

•Mostly based on the NOAEL in a suitable “mid-term“
study (28 days, 90 days, subchronic neurotoxicity,
developmental toxicity)
•Covers also exposures via dermal and inhalation routes
•If oral absorption (OA) is < 80 %, the AOEL must be
corrected for limited systemic bioavailability
•Systemic value

PoD
AOEL = × OA
UF
Currently no agreed EU guidance document available
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Reference values, AAOEL

Acute Acceptable Operator Exposure Level, given

in mg kg bw-1 d-1; is the reference value for non-
dietary 24 h exposure; it has to be compared to the
acute exposure of operators, bystanders and workers

•Needed for all active ingredients with evidence of

adverse effects after a single or few administrations
•Similar to ARfD
•Covers also exposures via dermal and inhalation routes
•If oral absorption (OA) is < 80 %, the AAOEL must be
corrected for limited systemic bioavailability
•Systemic value

PoD
AAOEL = × OA
Currently no agreed EU guidance document available
UF
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Risk assessment
Define the problem

Toxicological studies
Use (pattern)
and data

Reference doses Exposure

Risk

Communication
Use not safe: consider risk mitigation, where relevant
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Principle of Risk Assessment

Reference values (ADI, ARfD, AOEL, AAOEL) are

compared to the expected exposure (dietary and
occupational), taking into account all routes.

If a reference value is exceeded, either

the intended application is not safe,
or
risk mitigation methods need to be considered.

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GERMAN FEDERAL INSTITUTE

Thank you
Thank you
FOR RISK ASSESSMENT

German Federal Institute for Risk Assessment

Max-Dohrn-Str. 8-10  10589 Berlin, GERMANY
Phone +49 30 - 184 12 - 0  Fax +49 30 - 184 12 - 47 41
bfr@bfr.bund.de  www.bfr.bund.de/en
Further reading

OECD test guidelines:

http://www.oecd.org/env/ehs/testing/oecdguidelinesforthetestingofchemi
cals.htm

Text books on toxicology: e.g.,

Hayes: Principles and methods of toxicology
Jacobson-Kram & Keller: Toxicological testing handbook (Principles,
Applications, and Data Interpretation)

ECHA: Guidance on information requirements and chemical safety

assessment; Chapter R.7 (endpoint specific guidance)
https://echa.europa.eu/guidance-documents/guidance-on-information-
requirements-and-chemical-safety-assessment
Guidance on CLP (classification and labelling of products)
https://echa.europa.eu/guidance-documents/guidance-on-clp
Guidance on biocides legislation
https://echa.europa.eu/guidance-documents/guidance-on-biocides-
legislation
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Further reading

IPCS Harmonization project:

http://www.who.int/ipcs/methods/harmonization/en/
e.g., Risk Assessment Toolkit, Uncertainty in Hazard Assessment, Risk
Assessment Toolkit

Principles and methods for the risk assessment of chemicals in food

(Environmental Health Criteria 240)
http://www.who.int/foodsafety/publications/chemical-food/en/

Guidance documents on Acute Reference Dose and Acceptable Operator

Exposure Level
http://ec.europa.eu/food/plant/pesticides/approval_active_substances/gui
dance_documents_en

Page 40
 Illustration created with http://www.tagxedo.com
Who needs to be protected?

Foetuses
Toddlers
Children
Teenagers
Adults (men/women)
Elderly
Sick
Disabled
Pregnant women
People with specific vulnerabilities

Page 41
Focus on human health: Hazard – Risk – Exposure (I)

Illustration created with http://www.tagxedo.com

What are the toxic properties of a pesticide?

Acute toxicity
Skin and eye irritation / corrosion
Allergy
Mutation
Cancer
Malformations
Retardations
Fertility
Effect on lactation
Neurotoxicity
Organ damage
…

Page 42