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Administrative data

Key value for chemical safety assessment

Additional information

Table 7.6/1: Summary of genotoxicity tests

 

Test n°

Test / Guideline

Reliability

Focus

Strains tested

Metabolic activation

Test concentration

Statement

1

 

May, 2012

Ames Test

(OECD 471)

K, rel. 1

Gene mutation

TA 1535,

TA 1537,

TA 98,

TA 100,

WP2 uvrA

-S9

+S9

Up to 5000 µg/plate

-S9 : non mutagenic

+S9 : non mutagenic

2

 

Abramsson-Zetterberg, 2002

Ames Test

S, rel. 4

Gene mutation

TA97,

TA 98,

TA 100

-S9

+S9

-S9: up to 100 µg/plate

+S9: Up to 2000 µg/plate

-S9 : non mutagenic

+S9 : non mutagenic

3

 

Wild, 1983

Ames Test

S, rel. 4

Gene mutation

TA1535, TA100, TA1537, TA1538,

TA98

-S9

+S9

Up to 3600µg/plate

-S9 : non mutagenic

+S9 : non mutagenic

4

 

Woods, 2013

CHO/hprt test (OECD 476)

K, rel. 1

Gene mutation

Chinese hamster ovary cells

-S9

+S9

-S9: Up to 200 µg/plate

+S9: Up to 1600 µg/plate

-S9 : non mutagenic

+S9 : non mutagenic

5

 

Abramsson-Zetterberg, 2002

In vivo micronucleus

(eq. OECD 474)

K, rel. 2

Chromosomal aberration

Peripheral blood cells

NA

M: up to 1600 mg/kg bw

F: up to 1400 mg/kg bw

Non clastogenic, non aneugenic

 

Gene mutation Assays (Tests n° 1-4):

Three Bacterial Reverse mutation Assays (Ames test) were performed according or similarly to OECD 471 test guideline with Ethylene Brassylate (See Table 1). Test n°1 was selected as the key study. Test 2 was used as supporting data because only three strains of bacteria were used and Test 3 was used as supporting because results were not detailed. No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains in any test, with any dose of Ethylene Brassylate, either in the presence or absence of metabolic activation. The three tests indicate that Ethylene Brassylate does not induce gene mutations in bacteria whereas all positive control chemicals (with and without metabolic activation) induced significant increase of colonies. Ethylene Brassylate is therefore considered as non-mutagenic according to the Ames test.

Inability to produce gene mutation was confirmed in mammals using an in vitro forward mutation assay in Chinese hamster ovary cells (CHO/hprt test)(Test n°2). None of the dose levels up to the cytotoxicity limit of Ethylene Brassylate, either in the presence or absence of metabolic activation, induced significant mutant frequency increases in the initial or repeat tests. Ethylene Brassylate does not induce forward mutations at the hprt locus in CHO cells under activation and non activation conditions whereas both positive control chemicals (with and without metabolic activation) induced significant mutant frequency increases. Ethylene Brassylate is therefore considered as negative for inducing forward mutations at the hprt locus in CHO cells under activation and non-activation conditions used in this assay. This result confirms the results of the Ames test and extends the non-mutagenic effect of Ethylene Brassylate to mammalian cells.

 

Chromosomal aberration (Test n°5)

The clastogenic and aneugenic potential of Ethylene Brassylate was determined using an in vivo mammalian erythrocytes micronucleus assay (Test n°5), which identifies substances that cause micronuclei in erythroblasts. These micronuclei may originate from acentric fragments or whole chromosomes, and the test thus has the potential to identify both clastogenic and aneugenic chemicals. In this study erythroblasts were sampled from peripheral blood cells of mice. None of the i.p. dose levels of up to 1600 and 1400 mg/kg bw, in males and females respectively, induced increase in the frequency of micronucleated polychromatic erythrocytes (fMPCE), whereas the positive control chemical induced significant increases in the fMPCE. Ethylene Brassylate was therefore considered as negative for inducing chromosomal aberrations in mice bone marrow erythrocytes under the conditions used in this study. Ethylene Brassylate is therefore considered as non-clastogenic and non-aneugenic.

 


Justification for selection of genetic toxicity endpoint
No study was selected, since all in vitro and in vivo studies were negative

Short description of key information:
- Ames test: non mutagenic (OECD 471, GLP, K, rel. 1)
- hprt/CHO: non mutagenic (OECD 476, GLP, K, rel. 1)
- In vivo micronucleus: not clastogenic, not aneugenic (K, rel.2)

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008 including ATP3.

Self-classification:

Based on the available data, no additional classification is proposed.