2,2-dimethylpropane-1,3-diyl 2-ethylhexanoate

Genetic toxicity: in vitro

Administrative data

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:

20 Oct 1999 - 11 Nov 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP Monitoring Authority, Department of Health

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,2-dimethylpropane-1,3-diyl 2-ethylhexanoate

EC Number:

249-060-1

EC Name:

2,2-dimethylpropane-1,3-diyl 2-ethylhexanoate

Cas Number:

28510-23-8

Molecular formula:

C21H40O4

IUPAC Name:

2,2-dimethylpropane-1,3-diyl bis(2-ethylhexanoate)

Details on test material:

- Name of test material (as cited in study report): Neopentylglycol dioctanoate- Physical state: liquid, colourless- Lot/batch No.: 0709N- Storage condition of test material: RT, protected from light- Analytical purity: 98%

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of male rats treated with phenobarbitone/b-naphthoflavone

Test concentrations with justification for top dose:

Pretest: 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate without metabolic activationMain test: 50, 150, 500, 1500 and 5000 µg/plate with and without metabolic activation

Vehicle / solvent:

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

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-ethyl-N-nitro-N-nitrosoguanidine (ENNG; 3 µg/plate for TA 100 and 5 µg/plate for TA 1535); 9-aminoacridine (9AA; 80 µg/plate for TA 1537); mitomycin C (MMC; 0.5 µg/plate for TA 102); 4-nitroquinoline-N-oxide ( 4NQO; 0.2 µg/plate for TA 98)

Remarks:

without metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: aminoanthracene (2AA; 1 µg/plate for TA100 and 2 µg/plate for TA 1535 and TA 1537); benzo(a)pyrene (BP; 5 µg/plate for TA 98); dihydroxyanthraquinone (DANTHRON; 10 µg/plate for TA 102)

Remarks:

with metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)DURATION- Exposure duration: appr. 48 hNUMBER OF REPLICATIONS: triplicates in two independet experiments DETERMINATION OF CYTOTOXICITY- Method: relative total growth

Evaluation criteria:

The test material may be considered to be positive in this test system if the following criteria are met:The test material should have induced a reproducible, dose-related and statistically significant increase in the revertant count in at least one strain of bacteria.

Statistics:

Dunnett´s method of linear regression

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

at and above 1500 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS- Precipitation: Greasy, oily precipitate at and above 1500 µg/plate. This did not prevent the scoring of revertant coloniesRANGE-FINDING/SCREENING STUDIES: Yes- Test test material was non-toxic to the strain TA100. The test material formulation and the S9-mix used were both shown to be sterile.COMPARISON WITH HISTORICAL CONTROL DATA: Yes

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Mutagenicity on bacteria - Experiment I

 

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	TA102	TA98	TA1537
	Vehicle	99	14	303	28	15
-	50	99	14	300	31	11
-	150	110	22	318	27	18
-	500	114	21	311	29	14
-	1500	101	22	307	33	17
-	5000	111	18	300	26	14
Positive controls - S9	Name	ENNG	ENNG	MMC	4NQO	9AA
	Concentrations (μg/plate)	3.0	5.0	0.5	0.2	80
	Number of colonies/plate	492	428	979	143	455
+	Vehicle	100	13	315	37	18
+	8	89	12	329	28	16
+	40	97	13	338	31	16
+	200	96	11	336	29	17
+	1000	109	14	318	26	15
+	5000	98	13	320	35	13
Positive controls + S9	Name	2AA	2AA	DAN	BP	2AA
	Concentrations (μg/plate)	1.0	2.0	10	5	2
	Number of colonies/plate	753	228	939	161	556

 

ENNG = N-ethyl-N⁺-nitro-nitrosoguanidine

4NQO= 4-nitroquinoline-1-oxide

9AA = 9-Aminoacridine

2AA = 2-Aminoanthracene

BP = Benzo(a)pyrene

DAN = 1,8-Dihydroxyanthraquinone

 

 

 

Table 2: Mutagenicity on bacteria - Experiment II

 

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	TA102	TA98	TA1537
	Vehicle	114	24	283	36	9
-	50	100	22	279	34	8
-	150	122	23	302	35	13
-	500	120	28	306	32	14
-	1500	117	26	291	29	12
-	5000	121	28	278	28	10
Positive controls - S9	Name	ENNG	ENNG	MMC	4NQO	9AA
	Concentrations (μg/plate)	3.0	5.0	0.5	0.2	80
	Number of colonies/plate	465	370	885	173	1004
+	Vehicle	107	20	295	32	14
+	8	125	22	300	28	16
+	40	115	26	294	35	18
+	200	112	18	297	24	15
+	1000	116	22	321	29	20
+	5000	106	20	313	28	14
Positive controls + S9	Name	2AA	2AA	DAN	BP	2AA
	Concentrations (μg/plate)	1.0	2.0	10	5	2
	Number of colonies/plate	1791	156	649	201	469

ENNG = N-ethyl-N⁺-nitro-nitrosoguanidine

4NQO= 4-nitroquinoline-1-oxide

9AA = 9-Aminoacridine

2AA = 2-Aminoanthracene

BP = Benzo(a)pyrene

DAN = 1,8-Dihydroxyanthraquinone

Under the tested experimental conditions the test substance did not induce gene mutations in S. typhimurium strains up to the maximum dose. Therefore it is not considered to be mutagenic in this bacterial mutagenicity test in vitro.

Applicant's summary and conclusion

Conclusions:

it was concluded that 3-[(2-ethylhexanoyl)oxy]-2,2-dimethylpropyl 2-ethylheptanoate CAS: 28510-23-8 is not considered to be mutagenic based on the negative results obtained from the Ames test.

Executive summary:

A study was conducted to assess the mutagenic potential of 3-[(2-ethylhexanoyl)oxy]-2,2-dimethylpropyl 2-ethylheptanoate CAS: 28510-23-8. The test was done in an accredited GLP laboratory, in accordance with internationally recognised guidelines:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

EPA OPPTS 870.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

The bacterial reverse mutation assay was done so using numerous concentrations of the test substance, on S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102. Under the conditions of the study the 3-[(2-ethylhexanoyl)oxy]-2,2-dimethylpropyl 2-ethylheptanoate CAS: 28510-23-8 did not elicit a positive response, and as such it not considered to be mutagenic.