Reaction mass of aluminium chloride, iron dichloride, magnesium chloride and manganese dichloride

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

- genetic toxicity: For FeCl2 amd MnCl2 the data base is ambigous. None of the other ingredients of the test substance is deemed genotoxic, therefore no classification is needed.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

The test substance is a watery solution of metal chlorides and free hydrogenchloride.

The toxicity of this mixture has therefore to be regarded as a summary of the toxicity of the different ingredients. Due to the relative concentrations for genetic toxicity FeCl2, MnCl2, AlCl3 and HCl are regarded. MgCl2 the only other substance of high concentration is disregarded due to the low overall toxicity of this substance and as both chloride and magnesium ions are essential for cellular life and present in every cell in high abundance.

Summary:

For FeCl2 amd MnCl2 the data base is ambigous. None of the other ingredients of the test substance is deemed genotoxic, therefore no classification is needed.

The summary is based on the data presented below.

- in vitro:

FeCl2:

One fully reliable study on gene mutation in bacteria (Kim 2004) conducted according to GLP and OECD 471 is available showing a negative result. In the main study, iron dichloride did not increase reverse mutations of Salmonella tryphimurium (strains TA 98, TA 100, TA 1535 and TA 1537) and Escherichia coli (strain WP2 uvrA) with and without a metabolic activation system at 33.3, 100, 300, 1,000, 3,000 and 5,000 µg/plate. There was no statistically significant difference up to the maximum test concentration of 5,000 µg/plate (p >0.01). Precipitation was noted at doses greater than 300 µg/plate. It was concluded that iron dichloride did not exhibit mutagenic activity to any test strains under the test conditions.

The endpoint of cytogenicity is covered by an in vivo study (see below)

The endpoint gene mutation in mammalian cells is covered by read across from a respective study conducted with FeCl3 (Dunkel 1999, equivalent to OECD 476, Klimisch score 2). Studies in mammalian cells (L5178Y TK+/)showed that ferric chloride hexahydrate induced mutations in the presence of an induced rat liver S9 mix (Dunkelet al. 1999). The dose-related response occurred was accompanied by a marked increase in toxicity. The concentrations tested were0.2-1.2 µg Fe/mL with metabolic activation and 309-1030 µg/mL in the absence of S9 mix.

MnCl2:

Due to a delay in the correspondance between the registrant of this test substance and the Manganese Consortium, no first hand animal data is available.

The DRAFT TOXICOLOGICAL PROFILE FOR MANGANESE, 2008, by the Agency for Toxic Substances and Disease Registry creports (after assessment of in vitro and in vivo data is) that no clear conclusion is possible on the mutagenic potential of manganese compounds.

AlCl3:

All three studies are conducted with aluminium chloride, basic. As the test are conducted with buffered solutions and as chloride, hydroxide and sulfate are anions that are essential for cellular life and are found in living cells in high abundance their effect on toxicity is regarded negligible the results for this test item can be read across to aluminium trichloride (AlCl3).

One fully reliable study on gene mutation in bacteria (Verspeek-Rip 2010 A) conducted according to GLP and OECD 471 is available showing a negative result. S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvr A were tested up to the limit concentration and did not give a positive result.

One fully reliable study on in vitro mammalian cell gene mutation (Verspeek-Rip 2010 B) conducted according to GLP and OECD 476 is available showing a negative result. L5178Y mouse lymphoma cells were tested up to the limit concentration defined by the solubility of the test item and did not give a positive result.

One fully reliable study on in vitro cytogenicity (Buskens 2010, Micronucleus assay with human lyphocytes) conducted according to GLP and OECD 487 is available showing a negative result. Human lymphocytes were tested up to the limit concentration of 600 µg/mL defined by the solubility of the test item in and did not give a positive result.

A classification is not necessary.

HCl:

The results for three available, reliable mutagenicity tests (Isquith A - C) are negative. Positive results were seen in a mammalian cell gene mutation assay (Cifone 1987) and in a mammalian chromosomal abberation test. Nevertheless these results are due to the pH effects inflicted by hydrochloric acid that overcame the buffer capacities of the test systems. Hydrochloric acid rapidly dissociates almost completely in contact with water, releasing the chloride ion and the hydrogen ion which combines with water to form the hydronium ion. Both chlorine and hydronium ions are normally present in the body, and mammals constantly secrete gastric juices, containing hydrogen ion concentrations equivalent to 0.17 N HCl, into the stomach.

Hydrochloric is therefore deemed non-mutagenic in vitro. Accordingly no additional in vivo data is required.

- in vivo:

FeCl2:

The endpoint of cytogenicity is covered by an in vivo mammalian cell micronucleus test with FeCl2 in rats which was conducted in accordance with GLP and OECD 474 and was fully reliable (Ji 2004). The test was negative.

In addition FeCl2 was tested in a wing spot test with Drosophila melanogaster (Ogawa 1994), conducted in accordance with accepted scientific principles (Klimisch score 2). This is an in vivo mutagenicity test that detects mutations in somatic cells. trans-heterozygous larvae for the wing-hair mutations mwh and flr were orally treated with 1.1-4.5 g Fe/l at the third instar stage. Wings were inspected at the adult stage for spots expressing phenotypes of the markers. Ferrous chloride was found to be negative for the induction of wing-hair mutations under the conditions of the test.

MnCl2:

See above.

AlCl3:

See above.

HCl:

According to REGULATION (EC) No 1907/2006, in vivo genotoxicity assays will be required if a positive result is seen in in vitro genotoxicity assays.

Hydrochloric acid is not genotoxic in in vitro tests using bacterial or simple eukaryotic cells, while its effects on the pH of the medium precludes the possibility of testing in other in vitro non-bacterial systems.

Hydrochloric acid rapidly dissociates almost completely in contact with water, releasing the chloride ion and the hydrogen ion which combines with water to form the hydronium ion. Both chlorine and hydronium ions are normally present in the body, and mammals constantly secrete gastric juices, containing hydrogen ion concentrations equivalent to 0.17 N HCl, into the stomach.

Further tests on this compound are therefore not necessary; this data requirement is not triggered.

Justification for classification or non-classification

Based on the assessment of the genetic toxicity stated above, the test substance is deemed not to be genotoxic and correspondingly no classification is needed.