Estimated
testing number
costs REACH
Estimated
% of animal
REACH
2%
2.7%
1.9%
2%
2%
3%
2%
1.7%
3%
Developmental
Developmental toxicity
toxicity
Other
Other
Accumulation
Accumulation
Carcinogenicity
Ready
Biodegradation
12% 6.7%
4.6%
31%
3%
Skin
Skin sensitisation
sensitisation
Long
Long term
term repeated
repeated dose
dose
Short
Short term
term repeated
repeated dose
dose
Subchronic
Subchronic toxicity
toxicity
Cytogenicity
Cytogenicity mammalian
mammalian cells
cells
Further
Further mutagenicity
mutagenicity
Two-generation
Two-generation reproductive
reproductive
toxicity
toxicity
3%
7%
78%
8%
24%
For reprotox and dev tox around 21.7 Mio animals are needed
reprotox
and dev tox
between
if noFor
waiving
possibilities
willthe
be costs
taken may
into vary
account
(Annex IX)
about 700 million EURO and 1100 million EURO
Animal testing and alternative approaches for the human health risk assessment under the proposed new European
chemicals regulation
Thomas
Hoefer
et al. Arch
pp549-564;
Assessment of additional testing needs under REACH.
FinnTox.78;
Pedersen,
Jack de2004
Bruijn, Sharon M unn & Kees
van Leeuwen. EUR 20863 EN (2003) , http://ecb.jrc.it
In vitro tests under development
ES cellsFertilisation
for teratogenicity
Testicular
toxicity
Female
gamete
maturation
Hormonal
interaction
Reproductive cycle
Cross-cutting issues
post-natal
Development
Gamete
production
and
release
3 tests for steroidogenesis:
•Leydig cell line
•Granulosa cell line
Late Prenatal
Development
Early Prenatal
Development
Fertilisation
Implantation
tests culture
for gamete maturation:
•2
Follicle
3 tests for various target tissues:
primary
bovine
oocytes
6• tests
androgenic/estrogenic
5
tests
to
detect
testicular
toxicity :
4 tests to detect
toxicity during:
•Developmental
neurotox
• Follicle
culture
••The
CASA test
(motility of spermatozoa)
interaction:
The
fertilisation
•Developmental cardiotox
SCSA/Comet
(sperm
DNA evaluation
integrity)
chemicals
under
•10
2embryo
transcriptional
••The
•Toxicity
to thepre-implantation
skeletal assays
system (anti)estrogenic
and (anti)-androgenic
•Leydig
cells toxicity
Species:
Species:
•Sertoli
cells toxicity
•1 estrogenic
binding assay
••Mouse
•chemicals
1 androgenic
binding
assay
Bovine
under
consideration
••50
Human
10 chemicals tested
ReProTect I
ReProTect II
Development of in vitro tests
Proof of principles
Scientific objectives
-Placental toxicity
-Toxicity to the
uterus
Teratogenicity
Compilation of test
batteries to testing
strategies
Validation of building
blocks and testing batteries
Testing batteries for
teratogenicity
Preimplantation
embryo
-fertility
-testicular toxicity
Submission of tests to
Regulatory Authorities
for implementation into
regulatory decisions
Testing
batteries for
fertility
-female germ
cell toxicity
Validation of
estrogen receptor
binding tests
Endocrine
disrupters
1
Development of non-covered
sub-endpoints: e.g.:
hypothalamus-pituitary axis
2
3
4
6
5
time in years
7
8
9
10
Example: Testing strategy for REACH (ZEBET/ECVAM)
Has the substance already been
classif ied f or reproductive and
developmental toxicity – Cat 1, 2 ?
Stage 1
NO
Is the substance a genotoxic
Carcinogen (car. 1 or 2 and mut 3) or
germ cell mutagen (mut 1, 2) ? Have
appropriate risk management measures
been implemented
yes
yes
NO
Does the substance exhibit no
relevant human exposure?
yes
NO
In-Depth review of existing
toxicological database. A re data
adequate f or C & L risk
assessment/management?
Stage 2
No f urther testing
NO
A re there alerts f or reproductive
and/or developmental toxicity in
existing tox database and/or
signif icant exposure potential?
Stage 3
yes
positive
negative
Validated in vitro testing battery
covering the most prominent
endpoints e.g t eratogenicity,
f ertility, EDs
or
Screening f or
Reproductive/developmental
toxicity (OECD 421/422)
Validated in vitro test depending on
the nature of the alert and the
applicability domain
positive
negative
positive
negative
Negative outcome will be
conf irmed at next tonnage level;
cat 3 classification