Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

3 in vitro studies were available to estimate the mutagenic and clastogenic effect of the substance.

-       In the in-vitro mouse lymphoma mutation test (GLP compliant ), the 2-amino-4-hydroxyethylaminoanisole sulfate was considered mutagenic in the mouse lymphoma thymidine kinase locus assay when tested up to 500 and 100 µg/mL with and without metabolic activation respectively, using the cell line L5178Y.

-       In the bacterial reverse mutation test, the 2-amino-4-hydroxyethylaminoanisole sulfate (Lehmann Blau) when evaluated up to 5000 µg/plate was considered non-mutagenic in the Salmonella typhimurium reverse mutation assay, in the presence or absence of metabolic activation.

-       In the in vitro micronucleus test with cultured human peripheral blood lymphocytes, the  2-amino-hydroxyethylamino-anisole sulfate caused a statistically significant and biologically relevant increase in the frequency of micronuclei in the presence of S-9 mix and an ambiguous result in the absence of S-9 mix, when treatment was commenced 24 hours following mitogen stimulation.

Hence a tendancy of mutagenic properties of test substance was obserevd in the in vitro studies.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (positive)

Genetic toxicity in vivo

Description of key information

According to results of the key studies (OECD guideline 486, GLP comliant, 1991, Klimisch 1)(OECD Guideline 74, GLP compliant, 2002, Klimisch 2) the registered substance 2-amino-4-hydroxyethylaminoanisole sulfate did not induce mutagenic effect on in vivo genotoxicity test (micronucleus test and UDS test) on mice and rats.

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

Additional information

In in vitro genotoxicity studies, the test item showed mutagenic effects on mammalian cell lines (human lymphocytes on micronucleus test and L5178Y for TK locus selection test) but no effect was assessed on bacteria cells (Ames test). Indeed, 2-Amino-hydroxyethylamino-anisole sulfate induced a biologically relevant increase in mutations at the thymidine kinase locus in L5178Y cells. The registered substance induced chromosomal damage in an in vitro micronucleus test with cultured human peripheral blood lymphocytes from female donors. The test item caused a statistically significant and biologically relevant increase in the frequency of micronuclei in the presence of S9-mix, when treatment was commenced 24 hours following mitogen stimulation. In the absence of S9-mix, an equivocal result was obtained.

To determine potential genotoxicity of the substance, in vivo genotoxicity assays were performed : Micronucleus test and UDS assay. Under these experimental conditions, the substance did not showed genotoxic effect.

2-Amino-hydroxyethylamino-anisole sulfate did not induce chromosome aberrations or damage to the mitotic apparatus in bone marrow cells of mice after a single intraperitoneal administration under the test conditions used. Although there was no effect on the PCE/NCE ratio indicating bone marrow toxicity, systemic availability can be assumed after i.p. application and is further suggested by the general signs of toxicity observed in treated animals.

No dose level of 2-Amino-hydroxyethylamino-anisole sulfate revealed UDS induction in the hepatocytes of the treated animals as compared to the vehicle control. It can be stated that under the experimental conditions reported, 2-Aminohydroxyethylamino-anisole sulfate did not induce DNA damage that is detectable with the UDS test.

.

Justification for classification or non-classification

A battery of in vitro and in vivo genotoxicity tests was performed with 2-Aminohydroxyethylamino-anisole sulfate covering the relevant genetic endpoints. 2-Aminohydroxyethylamino-

anisole sulfate induced mutations in an in vitro mammalian cell gene mutation assay (MLA) The noted shift towards small colonies indicated a clastogenic potential. This is confirmed by the positive result of the in vitro micronucleus test with human lymphocytes. An in vivo micronucleus test using intraperitoneal administration and an in vivo / in vitro UDS test using oral application were negative and indicated that the

genotoxic potential seen in vitro is not expressed under appropriate in vivo test conditions.