Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
in vivo mammalian somatic cell study: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
GLP compliance:
yes (incl. QA statement)
Type of assay:
mammalian erythrocyte micronucleus test

Test material

Constituent 1
Chemical structure
Reference substance name:
1,2-Ethanediamine, N-(2-aminoethyl)-, reaction products with glycidyl tolyl ether
EC Number:
282-199-6
EC Name:
1,2-Ethanediamine, N-(2-aminoethyl)-, reaction products with glycidyl tolyl ether
Cas Number:
84144-79-6
Molecular formula:
C14H26N3O2
IUPAC Name:
1,2-Ethanediamine, N-(2-aminoethyl)-, reaction products with glycidyl tolyl ether
Test material form:
liquid: viscous
Details on test material:
Manufacturer Air Products and Chemicals
Batch # 8418406

Test animals

Species:
mouse
Strain:
ICR
Details on species / strain selection:
There was no marked difference in the toxicity of the test item between the sexes in the range finding study; therefore the main test was performed using only male mice.
Sex:
male
Details on test animals or test system and environmental conditions:
Sufficient albino Hsd: ICR (CD-1®) strain mice were obtained from Envigo, B.V., Horst, The
Netherlands. At the start of the main test the mice weighed 23.8 to 29.9 g and were
approximately six to ten weeks old. After a minimum acclimatization period of five days the
animals were selected at random and given a number unique within the study by tail marking
and a number written on a colour coded cage card.
The animals were housed in groups of five in solid-floor polypropylene cages with
wood-flake bedding. Free access to mains drinking water and food (Envigo Teklad 2014C
Global Certified Rodent Diet supplied by Envigo Laboratories UK Ltd., Oxon, UK) was
allowed throughout the study. Representative analyses of food and water quality are retained
in the laboratory archive.
The temperature and relative humidity were set to achieve limits of 19 to 25ºC and 30 to
70%, respectively. The rate of air exchange was approximately fifteen changes per hour and
the lighting was controlled by a time switch to give twelve hours light and twelve hours
darkness. The Delta Building Monitoring system was used during the course of the study.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
arachis oil
Doses / concentrationsopen allclose all
Dose / conc.:
1 000 mg/kg bw (total dose)
Remarks:
Range finfing study
Dose / conc.:
1 200 mg/kg bw (total dose)
Remarks:
Range finfing study
Dose / conc.:
1 600 mg/kg bw (total dose)
Remarks:
Range finfing study
Dose / conc.:
2 000 mg/kg bw (total dose)
Remarks:
Range finfing study
Dose / conc.:
1 200 mg/kg bw (total dose)
Remarks:
Final test
Dose / conc.:
600 mg/kg bw (total dose)
Remarks:
Final test
Dose / conc.:
300 mg/kg bw (total dose)
Remarks:
Final test

Examinations

Details of tissue and slide preparation:
Slide Preparation
Immediately following termination (i.e. 24 or 48 hours following dosing), both femurs were
dissected from each animal, aspirated with foetal bovine serum and bone marrow smears
prepared following centrifugation and re-suspension. The smears were air-dried, fixed in
absolute methanol, stained in May-Grünwald / Giemsa, allowed to air-dry and a cover slip
applied using mounting medium.
Evaluation criteria:
Slide Evaluation
Stained bone marrow smears were coded and examined blind using light microscopy at
x1000 magnification. The incidence of micronucleated cells per 4000 polychromatic
erythrocytes (PCE-blue stained immature cells) per animal was scored. Micronuclei are
normally circular in shape, although occasionally they may be oval or half-moon shaped, and
have a sharp contour with even staining. In addition, the number of normochromatic
erythrocytes (NCE-pink stained mature cells) associated with 1000 erythrocytes was counted;
these cells were also scored for incidence of micronuclei.
The ratio of polychromatic to normochromatic erythrocytes was calculated together with
appropriate group mean values and standard deviations.
Data Evaluation
Comparison was made between the number of micronucleated polychromatic erythrocytes
occurring in each of the test item groups and the number occurring in the vehicle control
group.
A positive mutagenic response is demonstrated when a statistically significant,
dose-responsive, toxicologically relevant increase in the number of micronucleated
polychromatic erythrocytes is observed for either the 24 or 48-hour kill times when compared
to the vehicle control group.
If these criteria were not fulfilled, then the test item was considered to be non-genotoxic
under the conditions of the test.
A positive response for bone marrow toxicity was demonstrated when the dose group mean
polychromatic to normochromatic ratio was shown to be statistically significantly lower than
the vehicle control group.
All data were statistically analysed using appropriate statistical methods as recommended by
the UKEMS Sub-committee on Guidelines for Mutagenicity Testing Report, Part III (1989).
The data was analysed following a transformation using Student's t-test (two tailed).

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Toxicity:
yes
Vehicle controls validity:
valid
Negative controls validity:
not applicable
Positive controls validity:
valid

Applicant's summary and conclusion

Conclusions:
The test item, 1,2-Ethanediamine, N-(2-aminoethyl)-, reaction products with glycidyl tolyl ether, was considered to be non-genotoxic under the conditions of the test.
Executive summary:

The test item, 1,2-Ethanediamine, N-(2-aminoethyl)-, reaction products with glycidyl tolyl ether, was considered to be non-genotoxic under the conditions of the test.