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Key value for chemical safety assessment

Genetic toxicity in vitro

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Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Jan - Feb 1992
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
E. coli WP2 strain or S. typhimurium TA 102 were not investigated.
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
E. coli WP2 strain or S. typhimurium TA 102 were not investigated.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
Information from study report:
- Name of test material: Emulgator E30, Fest
- CAS number: 5896-54-8
The CAS number in report (5896-54-8) is misleading, since this CAS number does only represent 1-Penta-decanesulfonic acid, sodium salt (1:1), but it is confirmed that the substance used is correctly assigned to CAS 68188-18-1.
- Appearance/Further information: The test substance was supplied as white wax-like leaf-let and was received at the test institute on March 06, 1991.
- Batch No. of test material: 210291
- Purity: 95 %
- Analytical reference: TGL 39237
- Solubility and stability of the test substance in the solvent/vehicle: The substance is known to be stable in water.
Target gene:
Histidine locus
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
S9 preparation from the liver of Aroclor 1254 induced Sprague-Dawley rats.
Test concentrations with justification for top dose:
For initial test and independent repeat: 5, 1.5, 0.5, 0.15, 0.05, 0.015, 0.005 mg/plate with and without metabolic activation
Vehicle / solvent:
Water
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-aminoanthracene 10 µg/plate (all strains)
Remarks:
The positive controls 2-nitrofluorene, Na-azide and 9-aminoacridine were used without S9 mix; the positive control 2-aminoanthracene was used with S9 mix.
Details on test system and experimental conditions:
METHOD OF APPLICATION: Plate Incorporation Test
For each concentration and tester strain three parallel plates were used.

DETERMINATION OF CYTOTOXICITY
- Method: background growth
Evaluation criteria:
The threshold value given in the following tables is the number of revertants (average number of revertant colonies from test compound) for which X² is >/= 6.6 with the average number of spontaneous revertant colonies being the corresponding solvent control. If the number of revertants with the test compound would be greater than the calculated threshold value the number of revertants with the test substance would be significantly greater than the spontanous rate in the solvent control (error probability p
Statistics:
The statistical significance of the increase in the mutation frequency is examined in the X²-test with respect to its statistical significance. A statistically significant dose-related increase in the number of revertants occurs if several dilutions of the test compound are significant greater than the average number of spontaneous revertant colonies.
Species / strain:
other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
Mutagenicity of the test material was not detected, neither in the initial experiment in concentrations up to and including 5 mg/plate, nor in the independent repetition with up to and including 0.5 mg/plate, both conducted with and without a metabolic activating system.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
In the first test toxic effects were exhibited when the highest concentration of 5 mg/plate was applied. The tester strains gave reduced numbers of revertants compared to the untreated control however, accompanied by a scattered background growth of the inoculated bacteria. Growth of strain TA 1538 was completely inhibited by a concentration of 0.5 mg/plate. Strain TA 1537 showed the same effect with a concentration of 1.5 mg/plate.
Therefore the highest concentration of the test substance was 0.5 mg/plate in the repetition experiments either with or without metabolic activation.
Executive summary:

A bacterial reverse mutation assay (Ames test) according to OECD TG 471 was carried out with and without a metabolic activating system with the 5 tester strains S. thyphimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100, however no E. coli WP2 strain or S. typhimurium TA 102 was investigated. One independent repetition of the test was performed. Mutagenicity of the test material was not detected. Toxicity to S. typhimurium could be observed at the higher concentrations.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
As explained in a separate document (Read Across Justification, Sadler, 2017; attached to this dossier, Chapter 13) both paraffin oils, sulfochlorinated, saponified and the read across substance are very similarly composed secondary alkane sulfonates (UVCBs); slight differences are caused by the production processes - for the paraffin oils, sulfochlorinated, saponified the sulfochlorination and for the read across substance the sulfoxidation. A comparative assessment of the available toxicological in formations confirms that there is no reason to deviate from the read across approach from a toxicological view. This assessment is contained in a separate document (Supporting information on human health assessment, Karthaus, 2017; attached to this dossier, Chapter 13).

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Informations given in (a) Read Across Justification, Sadler, 2017 and (b) Supporting information on human health assessment, Karthaus, 2017, both attached to this dossier, Chapter 13.

3. ANALOGUE APPROACH JUSTIFICATION
Informations given in (a) Read Across Justification, Sadler, 2017 and (b) Supporting information on human health assessment, Karthaus, 2017, both attached to this dossier, Chapter 13.

4. DATA MATRIX
Informations given in (a) Read Across Justification, Sadler, 2017 and (b) Supporting information on human health assessment, Karthaus, 2017, both attached to this dossier, Chapter 13.
Reason / purpose:
read-across source
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Executive summary:

Based on read across this result of a genetic toxicity study is relevant for paraffin oils, sulfochlorinated, saponified. As explained in a separate document (Read Across Justification, Sadler, 2017; attached to this dossier, Chapter 13) both paraffin oils, sulfochlorinated, saponified and the read across substance are very similarly composed secondary alkane sulfonates (UVCBs); slight differences are caused by the production processes - for the paraffin oils, sulfochlorinated, saponified the sulfochlorination and for the read across substance the sulfoxidation. A comparative assessment of the available toxicological informations confirms that there is no reason to deviate from the read across approach from a toxicological view. This assessment is also contained in a separate document (Supporting information on human health assessment, Karthaus, 2017; attached to this dossier, Chapter 13).

Here is the result of the in vitro genotoxicity study (HPRT) with the read across substance:

The test substance was tested in an in vitro gene mutation assay (HPRT) in V79 cells according to OECD TG 476. The selection of the concentrations was based on data from the pre-experiments, where higher doses showed cytotoxicity. In experiment I 55.0 μg/mL (with metabolic activation) and 8.25 μg/mL (without metabolic activation) were selected as the highest concentrations, in experiment II 54.4 μg/mL (with metabolic activation) and 104.0 μg/mL (without metabolic activation). Experiment II without metabolic activation was performed as a 20 h long-term exposure assay.

In both experiments no biologically relevant increase of mutants was found after treatment with the test item (with and without metabolic activation). No dose-response relationship was observed. The positive controls ethyl methanesulfonate and dimethylbenzanthracene induced distinct and biologically relevant effects in mutation frequency. Based on these results, the test item is considered to be non-mutagenic in the V79/HPRT-assay.

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

Genetic toxicity in vivo

Link to relevant study records
Reference
Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
As explained in a separate document (Read Across Justification, Sadler, 2017; attached to this dossier, Chapter 13) both paraffin oils, sulfochlorinated, saponified and the read across substance are very similarly composed secondary alkane sulfonates (UVCBs); slight differences are caused by the production processes - for the paraffin oils, sulfochlorinated, saponified the sulfochlorination and for the read across substance the sulfoxidation. A comparative assessment of the available toxicological in formations confirms that there is no reason to deviate from the read across approach from a toxicological view. This assessment is contained in a separate document (Supporting information on human health assessment, Karthaus, 2017; attached to this dossier, Chapter 13).

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Informations given in (a) Read Across Justification, Sadler, 2017 and (b) Supporting information on human health assessment, Karthaus, 2017, both attached to this dossier, Chapter 13.

3. ANALOGUE APPROACH JUSTIFICATION
Informations given in (a) Read Across Justification, Sadler, 2017 and (b) Supporting information on human health assessment, Karthaus, 2017, both attached to this dossier, Chapter 13.

4. DATA MATRIX
Informations given in (a) Read Across Justification, Sadler, 2017 and (b) Supporting information on human health assessment, Karthaus, 2017, both attached to this dossier, Chapter 13.
Reason / purpose:
read-across source
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Vehicle controls validity:
valid
Negative controls validity:
not applicable
Positive controls validity:
not applicable
Executive summary:

Based on read across this result of a genetic toxicity study is relevant for paraffin oils, sulfochlorinated, saponified. As explained in a separate document (Read Across Justification, Sadler, 2017; attached to this dossier, Chapter 13) both paraffin oils, sulfochlorinated, saponified and the read across substance are very similarly composed secondary alkane sulfonates (UVCBs); slight differences are caused by the production processes - for the paraffin oils, sulfochlorinated, saponified the sulfochlorination and for the read across substance the sulfoxidation. A comparative assessment of the available toxicological informations confirms that there is no reason to deviate from the read across approach from a toxicological view. This assessment is also contained in a separate document (Supporting information on human health assessment, Karthaus, 2017; attached to this dossier, Chapter 13).

Here is the result of the in vivo genotoxicity study ( Mammalian Erythrocyte Micronucleus Test) with the read across substance:

A mouse bone marrow micronucleus test was conducted similar to OECD TG 474 but at a time before the guideline was available. In this study 5 male and 5 female mice per dose group, received two oral (gavage) administrations, separated by 24 hours, at doses of 360, 720, and 1440 mg/kg bw (act. ingr.; corresponding to 600, 1200 or 2400 mg/kg bw test material).

In dose groups 360 and 720 mg/kg bw no effect on behaviour or general conditions was observed after test substance administration. 100 % mortality occurred in the highest dose group, therefore only bone marrow smears of the medium and low dose were assessed for genotoxicity. As result the numbers of polychromatic erythrocytes with micronuclei was unaffected by the treatment. The evaluation of the bone marrow smears revealed no indications for a test substance-related depression of erythropoesis, but the values of normochromatic erythrocytes scattered in a broad range for treated and control groups. Overall, no indication for a mutagenic potential was seen in this study.

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

Additional information

The test substance did not show genetic toxicity in an Ames test conducted according to OECD TG 471, with 5 tester strains, but without E. coli WP2 or S. typhimurium TA102, and in a mammilian cell gene mutation assay according to OECD TG 473. Both tests were conducted with and without a metabolic activating system.

An in vivo mammalian erythrocyte micronucleus test (OECD TG 474) likewise showed no indications for genotoxicity.

With respect to the missing strains S. typhimurium TA 102 and E. coli WP2 uvrA in the Ames: According to guideline OECD TG 471 these strains are recommended specifically to detect “oxidising mutagens, cross-linking agents and hydrazines”, none of which describe paraffin oil, sulfochlorinated, saponified.

Overall, the information on genetic toxicity is considered sufficient for assessment and no potential for a genotoxic potential is concluded for the substance.

Justification for classification or non-classification

According to Regulation (EC) No 1272/2008, Annex I, no classification is required for genetic toxicity.