Tetra-esterification products of C5-(branched and linear), C7, C8 and C10 fatty acids with pentaerythritol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions (no details on analytical purity given)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no analytical purity reported

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): only trade name given
- Analytical purity: no data
- Lot/batch No.: CRU 98433

Method

Target gene:

his operon (S. typhimurium) and trp operon (E. coli)

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of male Sprague Dawley rats treated i.p. with a single dose of 500 mg/kg bw Arochlor 1254

Test concentrations with justification for top dose:

Range-finding toxicity study (in TA 100 and WP2 uvrA): 6.67, 10.0, 33.3, 66.7, 100, 333, 667, 1000, 3330 and 5000 µg/plate, with and without metabolic activation
Main study (all strains): 33.3, 100, 333, 1000, 3330 and 5000 µg/plate, with and without metabolic activation

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

-S9: 2-NF (1 µg/plate, TA 98); SA (2 µg/plate, TA 100 and TA 1535); ICR-191 (2 µg/plate, TA 1537); 4-NQO (1 µg/plate, WP2 uvrA); +S9: BP (2.5 µg/plate, TA 98); 2-AA (2.5-25 µg/plate, TA 100, TA 1535, TA 1537 and WP2 uvrA)

Positive control substance:

4-nitroquinoline-N-oxide

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: 2-aminoanthracene; ICR-191

Remarks:

2-NF: 2-nitrofluorene; SA: sodium azide; 4-NQO: 4-nitroquinoline-N-oxide; BP: benzo(a)pyrene; 2-AA: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 52 ± 4 h

NUMBER OF REPLICATIONS: triplicates each in one experiment

DETERMINATION OF CYTOTOXICITY
- Method: inspection of bacterial background lawn

Evaluation criteria:

The results of the test were considered positive, if the following criteria were met:
- tester strains TA 98, TA 100 and WP2 uvrA: for a test article to be considered positive, it must produce at least a 2-fold increase in the mean revertants per plate of at least one of these tester strains over the mean revertants per plate of the appropriate vehicle control. This increase in the mean number of revertants per plate must be accompanied by a dose response to increasing concentrations of the test article.
- tester strains TA 1535 and TA 1537: for a test article to be considered positive, it must produce at least a 3-fold increase in the mean revertants per plate of at least one of these tester strains over the mean revertants per plate of the appropriate vehicle control. This increase in the mean number of revertants per plate must be accompanied by a dose response to increasing concentrations of the test article.

Statistics:

Mean values and standard deviations of revertants per plate were calculated.

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: in the absence of S9 mix, slight precipitation of the test substance was observed in all experiments at concentrations ≥ 100 µg/plate. In the absence of S9 mix, slight precipitates were noted at ≥ 333 µg/plate in the preliminary cytotoxicity study and at ≥ 1000 µg/plate in the main study.

RANGE-FINDING/SCREENING STUDIES: in a preliminary cytotoxicity study, the tester strains TA 100 and WP2 uvrA were treated with the test substance at concentrations ranging from 6.67 to 5000 µg/plate in the presence and absence of metabolic activation (S9 mix). No cytotoxicity was observed in these strains up to the limit dose of 5000 µg/plate, neither with nor without addition of S9 mix.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1. Test results of experiment (plate incorporation)

Bacterial Reverse Mutation Assay, mean revertant colonies/plate (mutation factor) (n=3 ± SD)
EXPERIMENT
S9-Mix	Without
Concentration (per plate)	TA 98	TA 100	TA 1535	TA 1537	WP2 uvrA
SC	25 ± 2	80 ± 9	14 ± 8	3 ± 1	26 ± 3
Test material
33.3 µg	19 ± 4	88 ± 5	12 ± 5	4 ± 4	19 ± 2
100 µg	21 ± 7	96 ± 12	11 ± 2	5 ± 4	19 ± 4
333 µg	22 ± 2	92 ± 9	13 ± 2	5 ± 5	23 ± 10
1000 µg	25 ± 6	97 ± 11	25 ± 2	5 ± 3	30 ± 3
3330 µg	22 ± 1	101 ± 3	10 ± 1	3 ± 2	32 ± 2
5000 µg	20 ± 6	85 ± 6	15 ± 3	4 ± 2	27 ± 3
PC
2-NF	206 ± 42	-	-	-	-
SA	-	549 ± 71	480 ± 3	-	-
ICR-191	-	-	-	277 ± 33	-
4-NQO	-	-	-	-	260 ± 43
S9-Mix	With
Concentration (per plate)	TA 98	TA 100	TA 1535	TA 1537	WP2 uvrA
SC	36 ± 2	102 ± 2	19 ± 1	7 ± 1	25 ± 5
Test material
33.3 µg	36 ± 7	93 ± 9	15 ± 2	8 ± 4	22 ± 5
100 µg	34 ± 7	97 ± 17	16 ± 4	9 ± 2	22 ± 6
333 µg	34 ± 6	87 ± 6	17 ± 2	9 ± 4	27 ± 3
1000 µg	39 ± 9	94 ± 17	17 ± 3	8 ± 2	26 ± 6
3330 µg	38 ± 8	92 ± 6	16 ± 4	6 ± 3	29 ± 11
5000 µg	34 ± 5	139 ± 5	20 ± 3	3 ± 2	24 ± 4
PC
BP	471 ± 14	-	-	-	-
2-AA	-	918 ± 296	124 ± 6	171 ± 32	278 ± 24
SC = Solvent control; PC = Positive control substances; SD = standard deviation; 2-NF: 2-nitrofluorene; SA: sodium azide; 4-NQO: 4-nitroquinoline-N-oxide; BP: benzo(a)pyrene; 2-AA: 2-aminoanthracene

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative