Dioctyl sebacate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Mutagenicity Testing of Di (2- ethylhexyl)phthalate and Related Chemicals in Salmonella

Author:

Errol Zeiger, Steve Haworth, Kristien Mortelmans, and William Speck

Year:

1985

Bibliographic source:

Environmental Mutagenesis 7:213-232 (1985)

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Bacterial reverse mutation assay was performed for the test chemical Di(2-ethylhexy1)adipate using Salmonella typhimurium strain TA100, TA1535, TA98, TA1537 with and without metabolic activation system

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Bis(2-ethylhexyl) adipate

EC Number:

203-090-1

EC Name:

Bis(2-ethylhexyl) adipate

Cas Number:

103-23-1

Molecular formula:

C22H42O4

IUPAC Name:

Bis(2-ethylhexyl) adipate

Details on test material:

- Name of test material: Di(2-ethylhexy1)adipate
- Molecular formula: C22H42O4
- Molecular weight: 370.57 g/mol
- Substance type: Organic
- Physical state: No data

Specific details on test material used for the study:

- Name of test material: Di(2-ethylhexy1)adipate
- Molecular formula: C22H42O4
- Molecular weight: 370.57 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: Analyzed purity: 97.6 %
- Impurities (identity and concentrations): No data

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

Liver homogenates (S-9 fraction) from Aroclor 1254-induced male Sprague-Dawley rats (RLI) or male Syrian hamsters (HLI)

Test concentrations with justification for top dose:

Lab 1: 0, 100, 333, 1000, 3333 or 10000 µg/plate
Lab 2: 0, 100, 333, 1000, 3333 or 10000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Lab 1: ET95 and Lab 2: DMSO
- Justification for choice of solvent/vehicle: Chemical solubility in respective solvents mentioned

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Lab 1: ET95 and Lab 2: DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 4-nitro-o-phenylenediamine (for TA98; without S9) and 2-amino-anthracene (2 AA; with S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: All tests were repeated atleast once

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data

Rationale for test conditions:

No data

Evaluation criteria:

A mutagenic response was defined as a reproducible, dose-related increase in the number of histidine-independent colonies over the spontaneous incidence; there was no requirement for a specific magnitude of increase.

Statistics:

No data

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA100, TA1535, TA98, TA1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: No data
- Other confounding effects: No data

RANGE-FINDING/SCREENING STUDIES: The final dose level selection was based on the results of a preliminary range-finding study conducted with TA100 in the presence and absence of S-9.

COMPARISON WITH HISTORICAL CONTROL DATA: No data

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: Mutagenic behavior of Di (2-ethylhexyl)adipate

Lab 1: Solvent ET95

Doseµg/plate	TA100						TA1535
	NA		10% HLI		10% RLI		NA		10% HLI		10% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	115	5.5	82	11.5	140	4.1	19	1.8	14	3.5	16	2.6
100	110	5.7	113	5.8	136	3.3	23	2.5	14	0.9	17	0.7
333	117	10.1	116	5.5	132	7.5	21	3.2	11	2.0	21	0.3
1000	119	6.4	107	5.3	128	5.9	20	1.7	19	2.4	20	1.9
3333	122	10.1	114	10.8	123	8.9	19	3.2	15	3.7	17	0.3
10000	109	13.0	108	15.1	129	6.7	17	3.8	14	2.2	19	1.3
Pos	2060	34.4	1093	6.8	1045	21.6	1331	34.8	108	8.7	64	4.5

 

Doseµg/plate	TA1537						TA98
	NA		10% HLI		10% RLI		NA		10% HLI		10% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	6	2.2	5	2.4	8	0.9	28	1.7	23	1.0	30	3.8
100	8	0.7	8	1.2	9	2.3	26	1.0	24	1.5	30	3.1
333	7	0.6	5	0.3	5	1.0	21	2.1	30	0.6	28	4.0
1000	8	3.1	9	1.7	7	1.2	27	1.2	30	4.6	34	2.3
3333	8	1.5	6	1.5	6	0.3	26	2.2	29	1.5	28	2.0
10000	6	1.5	7	1.8	6	2.4	24	3.8	31	1.5	25	2.7
Pos	422	92.9	111	7.0	122	1.5	1957	85.7	1008	13.3	847	33.9

 

Lab 2: Solvent DMSO

Doseµg/plate	TA100						TA1535
	NA		10% HLI		10% RLI		NA		10% HLI		10% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	81	11.2	133	8.7	93	4.2	9	2.3	14	0.6	9	1.7
100	91	10.5	84	4.1	90	6.7	8	0.3	8	0.0	11	3.0
333	79	3.2	94	9.1	80	3.7	8	0.9	5	0.3	10	2.5
1000	83	3.8	92	5.5	89	10.2	10	2.2	7	0.9	6	1.5
3333	74	3.8	89	10.1	86	2.8	11	2.6	5	0.9	7	2.7
10000	70p	6.3	86p	3.8	87p	3.2	11p	1.5	4p	0.6	5p	1.8
Pos	458	10.7	1161	10.3	553	5.5	375	4.9	315	3.8	271	15.5

 

Doseµg/plate	TA1537						TA98
	NA		10% HLI		10% RLI		NA		10% HLI		10% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	11	2.2	13	3.8	5	0.6	11	3.3	118	1.7	17	1.5
100	6	1.5	7	0.9	8	3.3	16	1.2	18	3.5	23	5.2
333	5	1.5	6	0.7	6	1.2	15	1.7	19	3.5	23	4.7
1000	9	2.0	8	0.9	7	1.2	10	1.8	18	1.5	21	3.9
3333	7	2.4	7	0.9	7	0.9	8	2.3	19	0.6	15	1.0
10000	6p	1.3	4p	0.3	7p	1.2	14p	2.7	16p	0.7	16p	0.9
Pos	63	7.8	348	4.0	311	10.5	498	19.7	864	46.7	320	18.8

Applicant's summary and conclusion

Conclusions:

Di(2-ethylhexy1)adipate did not induce mutation in the Salmonella typhimurium strain TA100, TA1535, TA98, TA1537 with and without rat and hamster liver S9 mix in both lab 1 and lab 2 study and hence is negative for gene mutation in vitro.

Executive summary:

Bacterial reverse mutation assay was performed for the test chemical Di(2-ethylhexy1)adipate using Salmonella typhimurium strains TA100, TA1535, TA98, TA1537 with and without rat and hamster liver S9 mix. The test chemical was dissolved in ET95 (lab 1) and DMSO (lab 2). The study was performed using the preincubation protocol at five dose levels of 0, 100, 333, 1000, 3333 or 10000µg/plate (lab 1 and 2) with incubation period of 48 hrs in the presence and absence of S9 mix.The final dose level selection was based on the results of a preliminary range-finding study conducted with TA100 in the presence and absence of S-9. No mutagenic response was noted for the test compound in the preliminary dose range finding study and the main study performed.

 

Di(2-ethylhexy1)adipate did not induce mutation in the Salmonella typhimurium strain TA100, TA1535, TA98, TA1537 with and without rat and hamster liver S9 mix in both lab 1 and lab 2 study and hence is negative for gene mutation in vitro.