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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
In order to evaluate the genetic toxicity potential of the registered substance three in vitro tests according to OECD guidelines were performed: i.e. Ames test (OECD 471), Chromosome aberration test (OECD 476) and HGPRT (OECD 473) test. All of these three in vitro tests were negative in result.
Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2014-07-22 to 2014-09-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Complete guideline conform study report available. Study is performed under GLP.
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay
Target gene:
The S. typhimurium histidine (his) reversion system measures his- → his+ .
The S. typhimurium are constructed to differentiate between base pair (TA 1535, TA 100) and frameshift (TA 1537, TA 98) mutations. E. coli WP2 uvrA 8pKM101) is constructed to diffentiate for base pair substitution.
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9 homogenate (Sodium phenobarbitone and beta-naphthoflavone induced)
Test concentrations with justification for top dose:
First trial : 0.0125, 0.025, 0.05, 0.1, 0.2 mg/plate (plate incorporation method)
confirmatory trial: 0.0125, 0.025, 0.05, 0.1, 0.2 mg/plate (preincubation method)
Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
solvent controls
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-Aminoanthracene; 4-nitroquinoline 1-oxide
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Remarks:
tested up to 0.2 mg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: other: bacteria
Remarks:
Migrated from field 'Test system'.

Table 1:   Emulsogen OG -Experiment I: (Plate incorporation test)

No. Revertants (mean of 3 plates)

Concentration

TA98

TA100

TA1535

TA1537

E. coli WP2 uvrA

[mg/plate]

- S9

+ S9

- S9

+ S9

- S9

+ S9

- S9

+ S9

- S9

+ S9

Negative Control

19.7

21.7

101.0

101.7

23.0

23.3

8.0

8.7

161.3

159.7

Solvent Control

21.3

20.3

102.3

102.0

23.7

22.3

8.3

7.7

160.7

161.3

Emulsogen OG

0.0125

21.0

20.3

100.7

101.0

22.0

21.7

7.3

8.0

161.3

161.0

0.025

21.3

19.3

101.3

101.7

22.0

22.0

8.0

7.3

160.7

161.3

0.05

20.0

21.3

100.7

101.3

22.7

21.7

7.7

7.7

161.7

159.7

0.1

20.3

21.0

102.3

101.7

22.3

22.0

8.0

8.3

160.3

160.3

0.2

20.7

21

101.3

101.7

22.7

21.3

8.0

7.7

160.3

160.7

Positive Control

420.7*

421.0*

391.7*

390.7*

143.7*

146.7*

123.3*

121.3*

385.0*

384.7*

 *= statistically significant than the solvent control group (p<0.05); Solvent control = DMSO;

Positive controls with S9

For allSalmonella typhimuriumstrains + S9: positive control = 2-aminoanthracene [4 µg /plate];

WP2uvrA +S9 positive control = 2-aminoanthracene [30.0 µg /plate];

Positive controls without S9

TA100 and TA1535-S9: positive control = sodium-azide [1.0 µg /plate];

TA1537-S9: positive control = 9-aminoacridine [50.0 µg /plate];

TA98-S9: positive control = 2-nitrofluorene [2.0 µg /plate];

WP2uvrA-S9: positive control = 4-nitroquinoline-N-oxide [5.0 µg /plate];

 

Table 2:       Emulsogen OG -Experiment II: (Pre-incorporation test)

No. Revertants (mean of 3 plates)

Concentration

TA98

TA100

TA1535

TA1537

E. coli WP2 uvrA

[mg/plate]

- S9

+ S9

- S9

+ S9

- S9

+ S9

- S9

+ S9

- S9

+ S9

Negative Control

21.3

21.0

101.3

102.3

22.3

22.3

8.3

7.3

162.3

161.7

Solvent Control

21.7

22.0

102.0

102.0

22.0

21.3

7.3

8.0

162.0

161.7

Emulsogen OG

0.0125

21.0

21.0

101.0

101.3

21.3

21.3

8.3

7.3

159.7

160.7

0.025

19.7

20.3

100.0

100.0

21.7

21.3

8.3

7.7

159.0

161.3

0.05

21.0

21.0

99.3

101.0

20.7

22.0

8.3

8.0

161.7

160.0

0.1

21.3

21.0

101.7

101.0

21.3

20.7

7.7

7.7

160.0

160.7

0.2

23.0

20.3

101.7

99.7

22.3

20.7

8.3

8.3

160.7

162.3

Positive Control

423.3*

423.0*

393.7*

391.7*

146.0*

144.7*

121.7*

122.7*

389.7*

387.3*

*= statistically significant than the solvent control group (p<0.05); Solvent control = DMSO;

Positive controls with S9

For allSalmonella typhimuriumstrains + S9: positive control = 2-aminoanthracene [4 µg /plate];

WP2uvrA +S9 positive control = 2-aminoanthracene [30.0 µg /plate];

Positive controls without S9

TA100 and TA1535-S9: positive control = sodium-azide [1.0 µg /plate];

TA1537-S9: positive control = 9-aminoacridine [50.0 µg /plate];

TA98-S9: positive control = 2-nitrofluorene [2.0 µg /plate];

WP2uvrA-S9: positive control = 4-nitroquinoline-N-oxide [5.0 µg /plate];

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

From the results obtained, the test item Emulsogen OG is found to be non mutagenic at and up to the highest concentration of 0.2 mg/plate in Bacterial Reverse Mutation test in the tester strain Solmonella typhimurium and Eschericia coli in the presence and the absence of metabolic activation.
Executive summary:

Test substance Emulsogen OG was evaluated for mutagenicity in a Bacterial Reverse Mutation Test according to OECD 471 Test guideline. The Salmonella typhimurium strains TA98, TA 100, TA1535, TA1537 and Eschericia coli WP2 uvrA (pKM101) strain were used as test system. Test item resulted in minimal precipitation at 0.2 mg/plate, hence initial cytotoxicity test was carried out with concentrations of 0.07, 0.08, 0.09.0.1 and 0.2 mg/plate. Based on the precipitation and initial cytotoxicity tests, the assay was performed at concentrations of 0.0125, 0.025, 0.05, 0.1 and 0.2 mg/plate with and without the metabolic actvation system S9. DMSO was used as solvent. Solvent control and appropriate positive controls were tested simultaneously. Two trials were carried out for this study in triplicates with plate incorporation method (Trial I) and preincubation method (Trial II).

From the experimental results obtained, the mean numbers of revertant colonies at the above mentioned concentrations were comparable to those of the DMSO in both trials with and without metabolic activation. There was no significant in increase in number of revertant colonies at any of the concetrations tested in both plate incorporation and preincubation method. The number of revertant colonies in the positive controls has shown 2.4 to 20.7 fold increase with statistical significance, at 5% level(p<0.05) under identical conditions.

From the results obtained, the test item Emulsogen OG is found to be non mutagenic at and up to the highest concentration of 0.2 mg/plate in Bacterial Reverse Mutation test in the tester strain Solmonella typhimurium: TA98, TA 100, TA1535, TA 1537 and Eschericia coli WP2 uvrA (pKM101) in the presence and the absence of metabolic activation.

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

Additional information

Additional information from genetic toxicity in vitro:

Justification for selection of genetic toxicity endpoint
Guideline conform GLP study, well documented report

Justification for classification or non-classification

Based on the available data no classification is warranted according to the criteria laid down in the EU Dangerous Substances Directive (67/548/EEC) and in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).