Ammonium acetate

Endpoint summary

Additional information

GENETIC TOXICITY IN VITRO:

Weight of evidence: Read-across approach from experimental data on analogues Sodium Acetate, Acetic Acid, Acetic anhydride, Phenoxy acetic Acid, Ammonia, Ammonium Sulfate, and Ammonia aqueous solution. Estimated data on Ammonium Acetate from Danish (Q)SAR database on mammalian cell gene mutation.

Read-across from experimental results obtained with Sodium Acetate:

In the first study, reported by Ishidate et al., 1984, a reverse mutation assay using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 was carried out with Sodium Acetate according to the method of Ames et al. (1975), but only with metabolic activation. No significant increases in the numbers of revertant colonies were detected in any S. typhimurium strains at the maximum dose tested. Applying the read-across approach, the substance Ammonium acetate is also considered as not mutagenic under test conditions.

In the same report, Ishidate et al. reported chromosomal aberrations tests with Sodium Acetate using a Chinese hamster fibroblast cell line, CHL. The cells were exposed to each sample at three different doses for 24 and 48 hours. No metabolic activation systems were applied. The incidence of cells with aberrations (including gaps) was 0%. Applying the read-across approach, the substance Ammonium acetate is also considered as not mutagenic under test conditions.

Read-across from experimental results obtained with Acetic Acid:

A test within the National Toxicology Program’s mutagenicity testing program and according to GLP was reported by Zeiger et al., 1992. This test was carried out with Acetic acid using Salmonella typhimurium strains TA 98, TA 100, TA 1535, and TA 97, with and without matabolic activation. Acetic acid did not show any mutagenic effect under test conditions. Based on these results, the read-across approach is applied and Ammonium acetate is also considered as not mutagenic under test conditions.

In the next report (by Morita et al., 1990) a cytogenetic assay was carried out with Acetic acid using Chinese hamster ovary K1 cells, with and without metabolic activation. In the absence of S9 mix, cells were exposed for 24 h to test substance at doses of 8, 10, 12, 14, and 16 mM. In the presence of S9 mix, cells were exposed for 6 h to test substance at doses of 4, 8, 10, and 12 mM, and recultured in fresh medium for 18 h. Cytotoxicity was evaluated by counting surviving cells.

The relationship between the pH of the medium and the clastogenic activity was examined. In order to study the effects of neutralization of the treatment medium, two kinds of treatment media were used. One was adjusted to pH 5.8 or pH 6.0 and the other was so adjusted and then immediately neutralized to pH 6.4 and pH 7.2 with 1 M NaOH. Acetic acid was not clastogenic at concentrations close to those showing cytotoxicity. Low pH did induce some artificial chromosome aberrations, but these were eliminated by neutralization of the test medium. The read-across approach is applied and Ammonium Acetate is also considered to be not clastogenic under test conditions.

Read-across from experimental results obtained with Acetic Anhydride:

In the paper reported by Seifreid et al. (2006), a L5178Y Mouse Lymphoma Cell Mutation Assay was performed with Acetic anhydride to test its mutagenic potencial. The chemical was tested with and without metabolic activation. The range of concentartions was 0.04 - 0.3 g/mL. The toxicity of test substance was also determined both with and without liver S9. The mutagenicity assay was performed according to the procedure described by Clive and Spector. Resistance to trifluorothymidine (TFT) was determined by adding TFT (final concentration, 3 µg/mL) to the cloning medium for mutant selection. Results have been evaluated under the traditional criteria (old evaluation) as well as the current international “harmonization” recommendations (new evaluation).

With old evaluation: Test substance was not mutagenic with metabolic activation, and was positive without metabolic activation (this positive result is not reliable, because full requeriments for a valid test were not met).

With new evaluation: Test substance was ambiguous with and without metabolic activation.

Based on these results, the read-across approach is applied and Ammonium acetate is considered to be ambiguous on mouse lymphoma cells, with and without metabolic activation.

Read-across from experimental results obtained with Phenoxy acetic acid:

A L5178Y Mouse Lymphoma Cell Mutation Assay was performed with Phenoxy acetic acid (National Toxicology Program Database). The chemical was tested with and without metabolic activation and, in general, tested concentrations were: 62.5, 125, 250, 500, 750, 1000, 1500, and 2000 µg/mL. Phenoxy acetic acid resulted to be not mutagenic with and without metabolic activation. It was toxic to cells, but at higher concentrations than precipitating concentrations.The read-across approach is applied and Ammonium acetate is considered to be not mutagenic on mouse lymphoma cells.

Read-across from experimental data on Ammonia, anhydrous:

A bacterial reverse mutation assay was performed with Ammonia with Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and TA 1538, and Escherichia coli WP2uvrA. The tested concentrations were: 500, 1000, 2500, 5000, 10000, and 25000 ppm. The test method was similar to OECD guideline 471. Ammonia was negative in an Ames test with and without metabolic activation. Applying the read-across approach, the substance Ammonium Acetate is also considered as not mutagenic under test conditions.

Read-across from experimental data on Ammonium Sulfate:

In the publication reported by Tuschy and Obe (1988), a mutagenic test with Ammonium sulfate was performed on Chinese Hamster Ovary cells. Various concentrations were tested: 0, 106, 211, and 423 mg/mL. Treatment of Chinese Hamster Ovary (CHO) cells with 3.2 M (423 mg/mL) ammonium sulfate in the absence of a metabolic activation system did not result in chromosomal aberrations. However, ammonium sulfate enhanced the frequency of chromosome type aberrations, which had been induced by the restriction endonuclease Alu I, but this is not indicative of a mutagenic effect of ammonium sulfate. Based on these results, and applying the read-across approach, the substance Ammonium Acetate is also considered as not mutagenic under test conditions.

In the publication reported by Obe and Kamra (1986), another mutagenic test with Ammonium sulfate was performed on Chinese Hamster Ovary cells. No chromosomal aberrations were found upon exposure of Chinese hamster ovary cells to ammonium sulfate. Based on these results, and applying the read-across approach, the substance Ammonium Acetate is also considered as not mutagenic under test conditions.

Read-across from experimental data on Ammonia, aqueous solution:

A bacterial reverse mutation assay was performed with Ammonia, aqueous solution and E. coli Sd-4-73, without metabolic activation. Aqueous ammonia was found to be not mutagenic in a bacterial reverse mutation assay without metabolic activation. Based on these results, and applying the read-across approach, the substance Ammonium Acetate is also considered as not mutagenic under test conditions.

Estimated results with Ammonium Acetate from Danish (Q)SAR Database:

 A Danish (Q)SAR prediction with the Multicase model was realized to estimate the mutagenic potencial of Ammonium acetate on mammalian cells (mouse lymphoma and HGRT (CHO): Chinese hamster ovary cell HGPRT forward mutation assay). The substance Ammonium acetate was predicted to be not mutagenic in mammalian cells. This prediction should be used for classification and risk assessment.

GENETIC TOXICITY IN VIVO:

Key studies: Read-across from experimental results obtained with Sodium Acetate:

The Testicular DNA-synthesis inhibition test (DSI test) was performed on male mice with Sodium Acetate. This is not a standard genotoxicity test system, but it provides evidence that acetic acid, sodium salt is not genotoxic in animals. The basis of the method is to measure 3H-thymidine incorporation. Animals receive a single oral dose by gavage at concentrations of 200, 500, and 1000 mg/kg bw of test substance. No inhibitory effect on DNA-replication was detectable in animals treated with Sodium Acetate. Based on this experimental data, and applying the read-across approach, the substance Ammonium acetate is also considered as not mutagenic under test conditions.

Short description of key information:

Genetic toxicity in vitro:

Weight of evidence: Ammonium Acetate is considered to be not mutagenic. Read-across approach from experimental data on analogues Sodium Acetate, Acetic Acid, Acetic anhydride, Phenoxy acetic acid, Ammonia anhydrous, Ammonium sulphate, and Ammonia aqueous solution. Data is provided on bacterial tests, chromosome aberration test and mammalian cell gene mutation assay. Estimated data on Ammonium Acetate from Danish (Q)SAR database on mammalian cell gene mutation.

Genetic toxicity in vivo:

Key studies: Read-across from experimental results with Sodium Acetate: The Testicular DNA-synthesis inhibition test (DSI test) was performed on male mice. No inhibitory effect on DNA-replication was detectable. The read-across approach was applied and the substance Ammonium acetate is also considered as not mutagenic.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on these results, Ammonium Acetate is not classified for mutagenicity.