Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

A registration of estradiol was already submitted earlier and is public available on the ECHA website. Chapter 7, which is still valid from today's perspective, was amended to fulfill the current information requirements. Consequently the migrated data (IUCLID 5 to IUCLID 6) was kept unchanged and only modified if there was a need for further information and/or to pass the technical completeness check (TCC).

None of the six tester strains S. typhimurium TA1535, TA100, TA1537, TA1538, TA98 and E.coli WP2uvrA showed increased reversion to prototrophy in assays with the test item at the doses tested between 0.05 and 5.0 mg/plate, either in the absence or presence of metabolic activation. Precipitates in the agar were found starting from 0.5 mg/plate onwards. Growth inhibition of the background lawn was observed in all strains at 2.5 and 5.0 mg/plate without and with metabolic activation.The counts recorded on appropriate negative control plates confirmed the characteristically spontaneous reversion rates of the tester strains. Furthermore, appropriate positive controls with known mutagens produced the expected distinct increase in the number of revertant colonies.

Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
February 2003
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
- Solubility and stability of the test substance in the solvent/vehicle: The solution and further dilutions were prepared immediately before addition to the test bacteria. Thus, no remarkable instability is expected which could influence the outcome of
the study.
Target gene:
Histidine gene locus
Species / strain / cell type:
S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538
Additional strain / cell type characteristics:
not applicable
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced male Sprague-Dawley rat liver S9 mix
Test concentrations with justification for top dose:
0.05 - 5.0 mg/plate (with/without S9 mix; plate incorporation and the experiment with preincubation)
Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
sodium azide
N-dimethylnitrosamine
benzo(a)pyrene
cyclophosphamide
ethylmethanesulphonate
other: 4-nitro-o-phenylenediamine, 2-aminoanthracene , N-Methyl-N´-nitro-N-nitrosoguanidine
Evaluation criteria:
A positive response was considered if the number of revertants of the compound groups compared to the number of revertants of the negative group was reproducibly higher than 2-fold. A dose-dependent increase in the number of
revertants was also considered to indicate a mutagenic effect.
Statistics:
The arithmetic means of the number of mutant colonies of the 3 parallel plates in the negative control groups were compared with those of the compound groups. However, in the tables the given mean values are rounded to the nearest integer.
Species / strain:
S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
Precipitates in the agar were found starting from 0.5 mg/plate onwards.
Generally, growth inhibition of the background lawn was observed at 2.5 and 5.0 mg/plate.
The preincubation experiment with and without S9 mix on strain TA 1537, however, was repeated in order to check the results of the positive control anthracene-2-amine.

None of the six tester strains S. typhimurium TA1535, TA100, TA1537, TA1538, TA98 and E.coli WP2uvrA showed increased reversion to prototrophy in assays with the test item at the doses tested between 0.05 and 5.0 mg/plate, either in the absence or presence of metabolic activation. Precipitates in the agar were found starting from 0.5 mg/plate onwards. Growth inhibition of the background lawn was observed in all strains at 2.5 and 5.0 mg/plate without and with metabolic activation.The counts recorded on appropriate negative control plates confirmed the characteristically spontaneous reversion rates of the tester strains. Furthermore, appropriate positive controls with known mutagens produced the expected distinct increase in the number of revertant colonies.

Executive summary:

The mutagenic potential of the test substance was evaluated in a Salmonella/microsome test with the S. typhimurium strains TA 98, TA 100, TA 1535, TA 1537, TA 1538 and E. coli WP2uvrA in the presence and absence of S9 mix according to OECD TG 471. Evidence of mutagenic activity was not seen up to the maximum recommended dose level of 5.0 µl/plate. No substantial increases in revertant colony numbers of any of the six tester strains were observed at any dose level in the presence and absence of metabolic activation. Therefore, the test substance was considered to be non-mutagenic in the Salmonella typhimurium reverse mutation assay.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

A registration of estradiol was already submitted earlier and is public available on the ECHA website. Chapter 7, which is still valid from today's perspective, was amended to fulfill the current information requirements. Consequently the migrated data (IUCLID 5 to IUCLID 6) was kept unchanged and only modified if there was a need for further information and/or to pass the technical completeness check (TCC).

It was reported that estradiol induces micronuclei in the bone marrow of mice after 0.1 to 10 mg/kg i.p. and chromosomal aberrations in human lymphocytes in vitro (Dhillon and Dhillon 1995). However, Ashby et al. were not able to reproduce the previously published positive findings in mice even when using 15 times higher doses of estradiol (up to 150 mg/kg i.p.) (Ashby et al. 1997). A limited micronucleus assay conducted in sexually immature, female rats also indicated that estradiol was inactive at a dose level that elicited a potent estrogenic response in the uterus of the treated animals (20 µg/kg i.p.). Furthermore, Shelby et al. also performed micronucleus assays with estradiol at doses up to 1250 mg/kg i.p. in male mice and male rats with negative results (Shelby et al. 1997). In a second step they tried to reproduce the findings reported by Dhillon and Dhillon in female mice and rats by using doses up to 10 mg/kg but again saw no increases in the frequency of micronucleus formation. Thus, the positive results reported by Dhillon and Dhillon could not be confirmed by other working groups.   The contribution of catechol metabolites generated from estradiol with their potential to cause oxidative stress and genetic damage in the process of tumorigenesis has been discussed in the literature (Yager and Liehr 1996). Much of this work has been performed in the context of the induction of renal tumors inmaleSyrian hamsters.However, the proposed genotoxic mechanism of tumor induction by estradiol is contradicted by the observation that it can be prevented by coadministration of progesterone or ethinylestradiol to male hamsters and it does not explain that tumor induction does not occur in intact female hamsters (Yager and Liehr 1996,Li et al. 1998).These observations shed doubt on the mechanistic predictivity of the male hamster kidney model with regard to its relevance human situation.Since the majority of results obtained in recognized test systems for mutagenicity testing to not support a mutagenic potential dor estradiol, this compound is classified as non-mutagenic according to Regulation (EC) 1272/2008 (CLP) .

Not classified according to German legislation (TRGS-905).

Lit: - Ashby J, Fletcher K, Williams C, Odum J, Tinwell H. Lack of activity of estradiol in rodent bone marrow micronucleus assays. Mutat Res 1997; 395: 83-88 - Dhillon VS, Dhillon IK. Genotoxicity evaluation of estradiol.Mutat Res 1995; 345: 87-95 - Li JJ et al.Prevention of estrogen carcinogenesis in the hamster kidney by ethinylestradiol: some unique properties of a synthetic estrogen. Carcinogenesis 1998; 19(3): 471-77 - Liehr JG.Is estradiol a genotoxic mutagenic carcinogen? Endocrine Reviews 2000; 21(1): 40-54 -Shelby MD, Tice RR, Witt KL.17ß-Estradiol fails to induce micronuclei in the bone marrow cells of rodents. Mutat Res 1997; 395: 89-90 -Yager JO, Liehr JG: Molecular mechanism of estrogen ¿ Carcinogenesis. Ann Rev Pharmacol Toxicol 1996; 36: 203-32

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

A registration of estradiol was already submitted earlier and is public available on the ECHA website. Chapter 7, which is still valid from today's perspective, was amended to fulfill the current information requirements. Consequently the migrated data (IUCLID 5 to IUCLID 6) was kept unchanged and only modified if there was a need for further information and/or to pass the technical completeness check (TCC).

In vitro

Test system

Substance

Test concentration

End point/Effect

Literature

pBR322
Plasmid
DNA
 
0.01-0.1 mmol/l
(-S9)
DNA damage
 
Single-strand breaks with
2 and 4-hydroxy estradiol
and estrone; negative with
estradiol and estrone
 
Yoshie & Ohshima, 1998. Free Radicals
Biol. Med., 15, 341-348 as cited by
Toxicological Evaluation of certain
Veterinary Drug Residues in Food. WHO
Food Additives Series: 43, prepared by the
Fifty-second meeting of the Joint
FAO/WHO Expert Committee on Food
Additives (JECFA), 2000.
S. typhimurium
TA100
TA1535,
TA1537,
TA1538,
TA98 and
E. coli WP2
Estradiol
valerate
2500 and 5000
µg/plate
Reverse mutation
 
Negative
Study Report No. A16094, Schering AG,
Evaluation of
Estradiol valerate in a bacterial reverse
mutation test (Ames-Test) using S.
typhimurium and E. coli, dated 26 Aug.
2004.
S. typhimurium
TA100
 
50-1500
µg/plate
Reverse mutation
 
Negative
Liehr et al., 1986 J. Steroid Biochem., 24,
353-356 as cited by Toxicological
Evaluation of certain Veterinary Drug
Residues in Food. WHO Food Additives
Series: 43, prepared by the Fifty-second
meeting of the Joint FAO/WHO Expert
Committee on Food Additives (JECFA),
2000.
S. typhimurium
TA100,
TA1535,
TA97, TA98
17ß-
Estradiol
0-10000 µg/plate
Reverse mutation
 
Negative
National Toxicology Program, AMES Test,
Study Nr.A36310, 1997
S. typhimurium
TA100,
TA1535,
TA97,
TA98
 
1-10 000 µg/plate -S9
 
1-1000 µg/plate +S9
 
Reverse mutation
 
Negative
Dhillon & Dhillon, 1995. Mutat. Res., 345,
87-95as cited by Toxicological Evaluation
of certain Veterinary Drug Residues in
Food. WHO Food Additives Series: 43,
prepared by the Fifty-second meeting of the
Joint FAO/WHO Expert Committee on
Food Additives (JECFA), 2000.
Mouse
lymphoma
L5178Y cells
Estradiol
valerate
2500 and 5000
µg/plate

No increase in mutant

colonies without and with

metabolic activation.

 

Cytotoxic effects were at

20 ¿g/mL (pulse

treatment) and 15 ¿g/mL

(continuous treatment)

seen without S9 mix and

at 60µg/mL with S9 mix

Study Report No. A15234, Schering AG,
Forward cell
mutation assay with Estradiol valerate at
the thymidine kinase locus (TK+/-)in
mouse lymphoma L5178Y cells, dated 26
Aug. 2004.
Rat
hepatocytes
(male)
Estradiol
Up to 10 µg/ml
No increased unscheduled
DNA synthesis

At cytotoxic dose.

Pharma Research Report A090, Schering
AG, Study to evaluate the potential of
estradiol to induce unscheduled DNA
synthesis in isolated rat hepatocytes in
vitro, dated 02 Oct.1992.
Rat
hepatocytes
(female)
Estradiol
1. Experiment:
0.5, 1.0, 5.0,
10.0 or 20.0
µg/ml
2. Experiment:
5.0, 10.0, 15.0,
20.0 or 25.0
µg/ml
 
No reproducible
concentration dependent
increased unscheduled
DNA synthesis up to the
highest dose.
Pharma Research Report A697, Schering
AG, Unscheduled DNA synthesis in
primary hepatocytes of female rats in vitro
with Estradiol, dated 05 Oct.1993.
Cultured
human
lymphocytes
 
1-100 µg/ml
Chromosomal aberration,
sister chromatid exchange
 
Positive
Dhillon & Dhillon, 1995. Mutat. Res., 345,
87-95as cited by Toxicological Evaluation
of certain Veterinary Drug Residues in
Food. WHO Food Additives Series: 43,
prepared by the Fifty-second meeting of the
Joint FAO/WHO Expert Committee on
Food Additives (JECFA), 2000.
Rodent cells
 
 
Aneuploidy, unscheduled
DNA synthesis
 
Positive

IARC Monographs on the Evaluation of the

Carcinogenic Risk of Chemicals to

Geneva: World Health Organization,

International Agency for Research on

Cancer, 1972-PRESENT. (Multivolume

work), V21 (1979).

Rodent cells
 
 
Mutagenicity, DNA
damage, sister chromatid
exchange
 
Negative

IARC Monographs on the Evaluation of the

Carcinogenic Risk of Chemicals to

Geneva: World Health Organization,

International Agency for Research on

Cancer, 1972-PRESENT. (Multivolume

work), V21 (1979).

Syrian
hamster   
embryo cells
 
 
0-10 µg/ml
(-S9)

Cell transformation,

numerical chromosomal

changes

 

Positive

IARC Monographs on the Evaluation of the

Carcinogenic Risk of Chemicals to

Geneva: World Health Organization,

International Agency for Research on

Cancer, 1972-PRESENT. (Multivolume

work), V21 (1979).

Syrian
hamster
embryo cells
 
 
0-10 µg/ml
(-S9)
Gene mutation,
chromosomal aberration,
sister chromatid exchange,
unscheduled DNA
synthesis
 
Negative
Tsutsui et al., 1987 Carcinogenesis, 11,
1715-1719 as cited by Toxicological
Evaluation of certain Veterinary Drug
Residues in Food. WHO Food Additives
Series: 43, prepared by the Fifty-second
meeting of the Joint FAO/WHO Expert
Committee on Food Additives (JECFA),
2000.
Chinese
hamster V79
cells
 
0-100 µmol/l
Microtubule disruption
 
EC50, 10 µmol/l
Aizu-Yokota et al., 1995. Cancer Res., 55,
1863-1868 as cited by Toxicological
Evaluation of certain Veterinary Drug
Residues in Food. WHO Food Additives
Series: 43, prepared by the Fifty-second
meeting of the Joint FAO/WHO Expert
Committee on Food Additives (JECFA),
2000.
Syrian
hamster
embryo cells
 
1 µg/ml (-S9)
Adduct formation Increase
with estradiol and 2- and 4-
hydroxy estradiol
Hayashi et al., 1996. Mol. Carcinog., 16,
149-156 as cited by Toxicological
Evaluation of certain Veterinary Drug
Residues in Food. WHO Food Additives
Series: 43, prepared by the Fifty-second
meeting of the Joint FAO/WHO Expert
Committee on Food Additives (JECFA),
2000.
Nonhuman
17-ß
Estradiol
 
Sister-chromatid exchange
(SCE)
 
No conclusion

EMICBACK/60360; JNCI J NATL

CANCER INST 75:575-580,1985 as cited

by GENETOX database, Beta-Estradiol, last

update 1998

Human
lymphocytes
17ß-
Estradiol
With metabolic
activation
Sister-chromatid exchange
(SCE)
 
Negative
EMICBACK/60103; MUTAT RES
156:199-218,1985; EMICBACK/50904;
CANCER RES
43:4114-4118,1983 as cited by GENETOX
database, Beta-Estradiol, last update 1998
Cultured
human
lymphocytes
 
0.05-75 µmol/l
Numerical chromosomal
changes
 
Positive
Schuler et al., 1998, Environ. Mol. Mutag.,
31, 263-273 as cited by Toxicological
Evaluation of certain Veterinary Drug
Residues in Food. WHO Food Additives
Series: 43, prepared by the Fifty-second
meeting of the Joint FAO/WHO Expert
Committee on Food Additives (JECFA),
2000.
Cultured
human
lymphocytes 
 
0.05-75 µmol/l
Chromosomal breakage
 
Negative
Schuler et al., 1998, Environ. Mol. Mutag.,
31, 263-273 as cited by Toxicological
Evaluation of certain Veterinary Drug
Residues in Food. WHO Food Additives
Series: 43, prepared by the Fifty-second
meeting of the Joint FAO/WHO Expert
Committee on Food Additives (JECFA),
2000.
Human cells
 
 
Micronucleus formation
 
Positive

IARC Monographs on the Evaluation of the

Carcinogenic Risk of Chemicals to

Geneva: World Health Organization,

International Agency for Research on

Cancer, 1972-PRESENT. (Multivolume

work), V21 (1979).

Human cells
17ß-
Estradiol
1 µg/ml

Aneuploidy, chromosomal

aberrations,

sister chromatid exchange

 

Negative

Serova & Kerkis, 1974Genetica, 10, 142-

149 as cited by IARC Monographs on the

Evaluation of the Carcinogenic Risk of

Chemicals to: World Health

Organization, International Agency for

Research on Cancer, 1972-PRESENT.

(Multivolume work), V21 (1979).

There is no evidence of genotoxic effects in non-mammalian systems. Some steroidal estrogens
can damage mammalian DNA and chromosomes (IARC 1999). The most frequently reported
effects include DNA adduct formation, cytogenetic alterations (e.g., chromosome and
chromatid breaks, micronuclei, sister chromatid exchange), and changes in chromosome
number (aneuploidy). Most of these effects have been demonstrated in various tests using
animal cells or cell-free systems. Studies with cultured human cell lines showed evidence of
chromosomal aneuploidy, DNA strand breaks, micronucleus formation, and sister chromatid
exchange. Among mammals, including humans estradio undergos phase I (aromatic
hydroxylation to catechol intermediates) and phase II metabolism (glucuronidation,
sulfonation, and O-methylation). The ratio of metabolic products depends on the target tissue,
species, strain, sex, and experimental conditions.
IARC Monographs,
Volume 72,
Hormonal
Contraception and
Post-Menopausal
Hormonal Therapy,
1999 as cited by
National Toxicology
Program, 2004.
Eleventh Report on
Carcinogens,
Estrogens, Steroidal.

in vivo

Test system

Substance

Application

Test concentration

End point/Effect

Literature

B6C3F1
Mice. Female
17ß-
Estradiol
Intraperitoneal
Injection
0.1 mg/kg, 1 mg/kg
or 10 mg/kg
Micronucleus
 
Negative
National Toxicology Program,
Micronucleus bone marrow &
peripheral blood studies,
A79652, 1996.
B6C3F1
Mice, male
17ß-
Estradiol
Intraperitoneal
Injection
312.5 mg/kg, 625
mg/kg or 1250
mg/kg
Micronucleus
 
Negative
National Toxicology Program,
Micronucleus bone marrow &
peripheral blood studies,
A48053, 1996.
B6C3F1
Mice, male
17ß-
Estradiol
Intraperitoneal
Injection
0.1 mg/kg, 1 mg/kg
or 10 mg/kg
Micronucleus
 
Negative
National Toxicology Program,
Micronucleus bone marrow &
peripheral blood studies,
A55232, 1996.
Han Wistar
Rat, 3 male
and 3 female
17ß-
Estradiol
Daily
administration
for 14 d
 
Gavage
2 mg/kg/d
Decreased body weight gain

 

Compound

specific DNA

adduct detected

only in male rat

liver.

 

No correlation

between DNA

adduct formation

and tumorigenic

potential.

Shering Research Report
No. AG18, DNA adduct
analysis in liver of male
and female rats after daily
intragastric administration of
chlormadinine acetate,
megestrol acetate,
drospirenone,
ethinylestradiol,
norethisterone acetate,
gestodene, estradiol or
progesterone over a period of
14 d, dated 23 Aug. 1996.
F344 Rats,
male
17ß-
Estradiol
Intraperitoneal
Injection
312.5 mg/kg, 625
mg/kg or 1250
mg/kg
Micronucleus
 
Negative
National Toxicology Program,
Micronucleus bone marrow &
peripheral blood studies, Study
Nr. A92447, 1996.
Sprague-
Dawley rats,
female, 
mammary
glands
4-
hydroxy-
estrone
Injection
200 nmol
32P-postlabelling
technique
 
Reaction with dG
and dA produced
a stable N2-dG or
N6-dA adduct
The authors
suggested the
formation of
depurinating
adducts via 3,4-
quinone followed
by misreplication
of unrepaired
apurinic sites
Stack et al., 1998 Adv.
Pharmacol., 42, 833-836 as
cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Syrian
hamsters
 
Intraperitoneal
Injection
2-150 mg/kg bw
Adduct formation
Increase in kidney
at 50 mg/kg bw;
no increase in
liver
Han & Liehr, 1994.
Carcinogenesis, 15, 997-1000
as cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Syrian
hamsters,
male
 
Intraperitoneal
Injection
50 mg/kg bw
Adduct formation
Increase in
kidney; no time
dependence
Han & Liehr, 1994.
Carcinogenesis, 15, 997-1000
as cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.

Syrian

hamsters

 
Intraperitoneal
Injection
100 mg/kg bw
Adduct formation
 
Increase in liver 
1-2 h after dosing
but not later
 
Han & Liehr, 1994.
Carcinogenesis, 15, 997-1000
as cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.

Syrian

hamsters

 
subcutaneous
implant
25 mg
Adduct formation
 
Increase in kidney
on day 3 but not
day 6; no hepatic
adducts;
substantial
differences in
adduct levels in
controls between
days 3 and 6
Han & Liehr, 1994.
Carcinogenesis, 15, 997-1000
as cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Syrian
hamsters,
male
 
Intraperitoneal
Injection
100 mg/d
for 3 days 
Adduct formation
 
Negative for
kidney with
estradiol and
2- and 4-hydroxy
estradiol
 
Han & Liehr, 1994.
Carcinogenesis, 15, 997-1000
as cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
NBL rat
 
 
Dose not reported
serum level 14
times that of
control
Adduct formation
 
Unidentified
adduct after 16
but not 8 weeks
of treatment
Han et al., 1995
Carcinogenesis, 16, 951-954 as
cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Mongrel
dogs
 
 
Dose not
reported 
Adduct formation
 
Decrease in
prostate adduct
level; increase in
carbonyl content
 
Winter & Liehr, 1996. Toxicol.
Appl. Pharmacol., 136, 211-
219 as cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Mouse bone
marrow
 
Intraperitoneal
injection,
singe dose
 
100-10 000
µg/kg bw
Micronucleus
formation, sister
chromatid
exchange
 
Positive at highest
dose
Dhillon & Dhillon, 1995.
Mutat. Res., 345, 87-95 as
cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Rat bone
marrow
 
Subcutaneous
injection ,
three daily
injections
20 µg/kg bw
Micronucleus
formation
 
Negative
Ashby et al., 1997. Mutat.
Res., 395, 83-88 as cited by
Toxicological Evaluation of
certain Veterinary Drug
Residues in Food. WHO Food
Additives Series: 43, prepared
by the Fifty-second meeting of
the Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Mouse bone
marrow
 
Intraperitoneal
Injection,
singe dose
10-10 mg/kg bw
Micronucleus
formation
 
Negative
Ashby et al., 1997. Mutat.
Res., 395, 83-88 as cited by
Toxicological Evaluation of
certain Veterinary Drug
Residues in Food. WHO Food
Additives Series: 43, prepared
by the Fifty-second meeting of
the Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Mouse and
rat bone
marrow
 
Intraperitoneal Injection
0.1-10 mg/kg bw
Micronucleus
formation
 
Negative
Shelby et al., 1997. Mutat.
Res., 395, 89-90 as cited by
Toxicological Evaluation of
certain Veterinary Drug
Residues in Food. WHO Food
Additives Series: 43, prepared
by the Fifty-second meeting of
the Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Male and
female mice
 
3 injections
310-1250 mg/kg bw
Frequency of
polychromatic
erythrocytes
 
Negative
Shelby et al., 1997. Mutat.
Res., 395, 89-90 as cited by
Toxicological Evaluation of
certain Veterinary Drug
Residues in Food. WHO Food
Additives Series: 43, prepared
by the Fifty-second meeting of
the Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Male Syrian
hamster
 
Intraperitoneal
Injection
5-150 mg/kg bw
Frequency of
polychromatic
erythrocytes
 
Negative
Han & Liehr, 1994.
Carcinogenesis, 15, 997-1000
as cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Male Syrian
hamster
kidney and
liver
 
Subcutaneous
injection
25 mg
DNA damage
Han & Liehr, 1994.
Carcinogenesis, 15, 997-1000
as cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Male Syrian
hamster
 
Infusion
250 µg/ d for 7 days
DNA damage
Han & Liehr, 1994.
Carcinogenesis, 15, 997-1000
as cited by Toxicological
Evaluation of certain
Veterinary Drug Residues in
Food. WHO Food Additives
Series: 43, prepared by the
Fifty-second meeting of the
Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
NBL rat
 
Subcutaneous
capsules
16 weeks; dose
not reported
DNA damage
 
Single-strand
breaks in prostate
with estradiol +
testosterone
Ho & Roy, 1994. Cancer 
Lett., 84, 155-162 as cited by
Toxicological Evaluation of
certain Veterinary Drug
Residues in Food. WHO Food
Additives Series: 43, prepared
by the Fifty-second meeting of
the Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Male Syrian
hamster
 
Subcutaneous
capsules
20 mg via
subcutaneous
capsule
Chromosomal
aberration
 
Positive in kidney
 
 
Banerjee et al., 1994. Mutat.
Res., 311, 191-197 as cited by
Toxicological Evaluation of
certain Veterinary Drug
Residues in Food. WHO Food
Additives Series: 43, prepared
by the Fifty-second meeting of
the Joint FAO/WHO Expert
Committee on Food Additives
(JECFA), 2000.
Rat, male
17ß-
Estradiol
 
 
Sperm
morphology
 
Positive

Panel Report:

EMICBACK/50126. Mutat.

Res. 115:1-72,1983 as cited by

GENETOX database, Beta-

Estradiol, last update 1998

Mouse
17ß-
Estradiol
 
 
Sperm
morphology
 
Positive

Panel Report:

EMICBACK/50126. Mutat.
Res. 115:1-72,1983 as cited by
GENETOX database, Beta-
Estradiol, last update 1998



Short description of key information:
There is a large amount of in vitro as well as in vivo studies reported in the literature for various endpoints. Also mostly negative, there is inconclusive data. There was no reverse mutation in the Ames-test in different bacteria (No. A16094), but single strand breaks were detected in pBR322 Plasmid DNA at 0.1 mmol/l and Aneuploidy and unscheduled DNA synthesis occurred in rodent cells. At the same time, these results are opposed by negative results for Gene mutation, chromosomal aberration, sister chromatid exchange and unscheduled DNA synthesis. Unfortunately the test concentrations are not always documented and therefore interpretation is aggravated.
DNA adducts were found in vitro as well as in vivo and it is suggested that the formation of depurinating adducts via 3,4-quinone followed by misreplication of unrepaired apurinic sites. Daily oral administration of 2 mg/kg Estradiol for 14 d to Wistar rats resulted in a compound specific DNA adduct detected only in male rat liver. No correlation between DNA adduct formation and tumorigenic potential was detected (AG18).
There was no micronucleus formation in vivo, but chromosomal aberrations occurred and there was an effect on s perm morphology.

Endpoint Conclusion:

Justification for classification or non-classification

Due to the results of the key study no classification is required according to Regulation (EC) 1272/2008/EC (CLP), Annex I.