Reaction products resulting from the esterification of C18 unsaturated fatty acid with glycerol and glycerol oligomers

Endpoint summary

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

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Test item: Emulsogen OG
CAS No.: 86088-80-4
Batch No.: ESD0017954
Appearance: Yellow liquid
Purity: UVCB substance, active content 99.1 %(w/w)

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).