Fatty acids, tall-oil, esters with dipentaerythritol

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 from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

23 Aug - 1 Sep 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Comparable to guideline study with acceptable restrictions (no E.coli strain or equivalent S. typhimurium strain was tested; lack of data on test substance, methodological deficiencies, no bacteria were added to the plates for tester strain TA1538 in the repeat experiment)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(no E.coli strain or equivalent S. typhimurium strain was tested; lack of data on test substance, methodological deficiencies, no bacteria were added to the plates for tester strain TA1538 in the repeat experiment

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

189200-42-8

Cas Number:

189200-42-8

IUPAC Name:

189200-42-8

Details on test material:

- Name of test material (as cited in study report): only trade name given
- Physical state: pale yellow liquid
- Analytical purity: no data
- Storage condition of test material: at room temperature

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

other: S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

Cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from livers of Sprague-Dawley rats treated with Aroclor 1254.

Test concentrations with justification for top dose:

First experiment (Initial assay): 0.5, 5, 50, 500, 5000 µg/plate with and without metabolic activation
Second experiment (Repeat assay): 50, 100, 500, 1000, 5000 µg/plate with and without metabolic activation

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: acetone for the test substance, DMSO for the positive control substances

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

acetone, DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: -S9: 9-Aminoacridine (9AA; 100 µg/plate) for TA1537; N-Methyl-N-Nitro-N-Nitrosoguanidine (MNNG; 10 µg/plate) for TA100 and TA1535; 2-nitrofluorene (2NF; 5 µg/plate) for TA98 and TA1538; +S9: 2-Aminoanthracene (2AA; 2.5 µg/plate) for all strains

Remarks:

test substance was dissolved in acetone; positive control substances were dissolved in DMSO

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: Triplicates each in two independent experiments with one exception: in the repeat assay, inadvertently no TA 1538 bacteria were added to the plates at 50, 100 and 500 µg/plate without S9-mix.

DETERMINATION OF CYTOTOXICITY
- Method: notable reduction of the bacterial background lawn and/or a greater than 50% reduction in the mean number of revertants in comparison to the vehicle control

Evaluation criteria:

The test substance may be considered positive in this test system if the following criteria are met: For the test item to be considered mutagenic, three-fold (or more) increases in mean revertant numbers must be observed in comparison with vehicle control plates.
In addition, there must be evidence of a dose-related increase in the number of revertants over at least three concentrations of the test substance. All cultures must demonstrate the characteristic mean number of spontaneous revertants in the vehicle controls.
A lack of response in the positive controls or spontaneous revertant frequencies which were not in keeping with historical laboratory values would render that portion of the test invalid.

Statistics:

Mean values and standard deviation were calculated.

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation of the test substance was observed in the initial assay and the repeat assay.
- Other confounding effects: Beading of the test substance was observed in the initial assay and the repeat assay at 500 µg/plate and higher with and without metabolic activation in all strains. The only exception was the strain TA 100 in the repeat assay without metabolic activation, where a beading of the test substance was observed at 1000 and 5000 µg/plate.

RANGE-FINDING/SCREENING STUDIES: A range-finding test was conducted to determine the doses used for the main studies. Therefore, concentrations of the test substance of 0.5, 1, 5, 10, 50, 100, 500, 1000, 2000 and 5000 µg/plate were tested. Toxicity was not observed at any concentration tested. Precipitation of the test substance was observed at 1000 and 2000 µg/plate with metabolic activation and between 1 and 500 µg/plate without metabolic activation. Beading of the test substance was observed at 500 µg/plate and higher with metabolic activation and at 100 µg/plate and higher without metabolic activation.

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 the initial assay

Bacterial Reverse Mutation Assay, mean revertants colonies/plate (mutation factor)
INITIAL ASSAY (Plate Incorporation Test)
S9-Mix	Without
Test item (µg/plate)	TA98	TA 100	TA 1535	TA 1537	TA 1538
Negative control	17.7	105.7	10.0	3.0	5.3
Acetone	17.3	97.0	6.3	3.3	8.0
0.5	14.7	98.7	11.7	5.0	5.7
5	10.0	109.0	12.3	6.7	8.0
50	14.3	101.3	7.7	4.3	7.7
500	18.7 B	122.3 B	12.3 B	3.3 B	4.0 B
5000	16.0 B	114.3 B	11.3 B	5.3 B	4.7 B
DMSO	13.0	112.0	9.3	4.0	3.3
9AA	---	---	---	674.0*	---
MNNG	---	945.0*	1089.7*	---	---
2NF	595.0*	---	---	---	826.7*
S9-Mix	With
Test item (µg/plate)	TA98	TA 100	TA 1535	TA 1537	TA 1538
Negative control	22.3	111.7	9.3	4.0	12.3
Acetone	20.3	119.7	11.7	6.3	7.0
0.5	18.3	126.0	12.3	4.7	9.3
5	17.0	124.3	7.3	6.0	13.0
50	15.7	129.0	11.0	5.0	5.3
500	25.7 B	121.7 B	10.7 B	4.3 B	9.7 B
5000	18.0 B	116.3 B	8.7 B	5.3 B	14.7 B
DMSO	13.7	105.0	12.0	5.3	13.7
2AA	338.3*	911.3*	113.7*	31.7*	471.3*
9AA: 9-aminoacridine (100 µg/plate) MNNG: N-methyl-N-nitro-N-nitrosoguanidine (10 µg/plate) 2NF: 2-nitrofluorene (5 µg/plate) 2AA: 2 -aminoanthracene (2.5 µg/plate)
*, p<0.05; **, p<0.01 B: test substance beaded on the plate

Table 2: Test results of the repeat assay

Bacterial Reverse Mutation Assay, mean revertants colonies/plate (mutation factor)
REPEAT ASSAY (Plate Incorporation Test)
S9-Mix	Without
Test item (µg/plate)	TA98	TA 100	TA 1535	TA 1537	TA 1538
Negative control	14.0	120.0	9.3	4.7	7.0
Acetone	16.7	102.7	10.7	5.3	3.7
50	16.7	117.7	13.7	5.7	0#
100	15.7	108.3	10.0	7.3	0#
500	21.7 B	99.0	12.0 B	5.7 B	0#
1000	16.7 B	125.3 B	7.7 B	4.0 B	8.3 B
5000	15.0 B	112.3 B	9.0 B	3.3 B	4.3 B
DMSO	16.3	101.7	10.7	3.7	6.3
9AA	---	---	---	544.3*	---
MNNG	---	1644.3*	1341.3*	---	---
2NF	708.3*	---	---	---	1019.3*
S9-Mix	With
Test item (µg/plate)	TA98	TA 100	TA 1535	TA 1537	TA 1538
Negative control	24.0	108.0	7.3	6.0	16.0
Acetone	21.0	123.7	7.0	4.7	14.7
50	25.3	109.3	12.7	5.0	10.0
100	30.3	110.7	7.0	6.3	17.3
500	27.7 B	104.3 B	15.3 B	5.7 B	16.7 B
1000	24.7 B	110.3 B	9.7 B	11.0 B	19.3 B
5000	22.3 B	121.7 B	11.3 B	4.7 B	13.3 B
DMSO	17.3	110.0	9.0	7.0	14.0
2AA	270.3*	738.3*	70.0*	36.7*	241.3*
9AA: 9 -aminoacridine (100 µg/plate) MNNG: N-methyl-N-nitro-N-nitrosoguanidine (10 µg/plate) 2NF: 2-nitrofluorene (5 µg/plate) 2AA: 2 -aminoanthracene (2.5 µg/plate)
*, p<0.05; **, p<0.01 B: test substance beaded on the plate #: bacteria were inadvertently not added

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative