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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1992-01-20 - 1992-03-02
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Version / remarks:
1984
Deviations:
no
GLP compliance:
yes
Type of assay:
mammalian cell gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Isophorone diamine of Hüls AG, purity 99.9 %, produced 25 Oct 1991; ID No. 3630/81 365

Method

Target gene:
hypoxanthine-guanine phosphoribosyl-transferase (HPRT) locus
Species / strain
Species / strain:
Chinese hamster Ovary (CHO)
Details on mammalian cell lines (if applicable):
CHO (Chinese hamster ovary) K1 cells
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced Wistar rat liver S9
Test concentrations with justification for top dose:
0 - 2 mg/ml
Vehicle:
HO medium, dimethyl sulfoxide (DMSO)
Controls
Negative controls:
yes
Remarks:
= solvent/ vehicle control
Solvent controls:
yes
Remarks:
Medium
True negative controls:
no
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
Remarks:
positive control with metabolic activation: 3-Methylcholanthrene

Migrated to IUCLID6: without metabolic activation
Details on test system and conditions:
HGPRT assay. ADMINISTRATION:
- Dosing:
preliminary toxicity test:
0; 0.02; 0.03; 0.06; 0.12; 0.2; 0.3; 0.6; 1.2; 2.0 mg/ml
main study; 0; 0.02; 0.06; 0.2; 0.6; 2.0 mg/ml
- Number of replicates: 2
- Application: 2E+05 cells/25 cm2 flask; exposure time 4 h
- Positive and negative control groups and treatment:
positive, without metabolic activation:
300 ug ethyl methanesulfonate (EMS)/ml in HO medium
positive, with metabolic activation:
10 ug 3-methylcholanthrene (MCA)/ml in dimethyl sulfoxide
negative: HO medium (with / without S9 mix)
Evaluation criteria:
statistically significant, dose related increase in mutant frequency at concentrations of the test substance resulting in > 20 % cell survival. Mean
frequency > maximum spontaneous frequency
Statistics:
t-test

Results and discussion

Test results
Species / strain:
Chinese hamster Ovary (CHO)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Remarks:
none over test concentration range
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: strain/cell type: CHO (Chinese hamster ovary) K1 cells
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

no further remarks

Applicant's summary and conclusion

Conclusions:
In this HPRT test with Chinese Hamster ovary (CHO) cells according to OECD TG 476 (1984), Isophorone diamine concentrations of
20 - 2,000 mg/l (+/- S9 mix from Aroclor 1254 induced rat livers) did not significantly increase the mutant frequency of treated cells. Cytotoxicity was not observed. It is concluded that Isophoron diamine, in the presence as well as in the absence of S9 mix, demonstrates no mutagenic potential in this in vitro mammalian cell assay.
at any of the concentrations tested.
Executive summary:

The test item Isophorone diamine was tested for its ability to induce forward mutation at the hypoxanthine-guanine phosphoribosyl-transferase (HPRT) locus in Chinese hamster ovary (CHO) cells in vitro both in the presence and the absence of exogenous metabolic activation(by Arochlor 1254 -induced rat liver S9). The substance was tested in concentrations of 20 mg/l to the solubility limit of 2000 mg/l with and without S9 mix. In this concentration range, Isophorone diamine did not induce any detectable cytotoxicity. Compared to the negative control treatment with Isophorone diamine in both assays (with and without metabolic activation) did not result in any reproducible statistically or biologically significant increase of the mutation frequency of the HPRT locus. It is concluded that Isophoron diamine, in the presence as well as in the absence of S9 mix, demonstrates no mutagenic potential in this in vitro mammalian cell assay.