Diisopropyl adipate

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:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1996

Reference 2

Reference Type:

secondary source

Title:

Dibutyl adipate CAS N°: 105-99-7

Author:

OECD

Year:

1996

Bibliographic source:

SIDS Initial Assessment Report for SIAM 4; Tokyo, Japan, 20-22 May 1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(adopted in 1983)

Deviations:

no

Remarks:

the current guildeline - adopted in 1997 - requires other positive controls +S9 beside 2-aminoanthracene

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Version / remarks:

(adopted in 1983)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Dibutyl adipate

EC Number:

203-350-4

EC Name:

Dibutyl adipate

Cas Number:

105-99-7

IUPAC Name:

dibutyl adipate

Details on test material:

- Name of test material (as cited in study report): Dibutyl adipate
- Analytical purity: 99.8%
- Lot/batch No.: N-41001
- Storage condition of test material: At room temperature

Method

Target gene:

his operon and trp operon

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 rats treated with phenobarbital and 5,6-Benzoflavone.

Test concentrations with justification for top dose:

312.5, 625, 1250, 2500 and 5000 µg/plate

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: -S9: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (0.01 µg/plate) for TA100, WP2uvrA, (0.1 µg/plate) for TA98; sodium azide (0.5 µg/plate) for TA1535; 9-aminoacridine (80 µg/plate) for TA1537; +S9: 2-aminoanthracene (up to 10 µg/plate) all strains

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

DETERMINATION OF CYTOTOXICITY
- Method: Inspection of the bacterial background lawn,
In a preliminary experiment employing all five applied tester strains triplicates of controls and single plates with test substance (50, 150, 500, 1500 or 5000 µg/plate) were incubated to determine cytotoxicity in the presence or absence of S9.

Evaluation criteria:

If the number of colonies with revertants was as double as that of control group and dose-related response was observed, the interpretation of the result was positive.

Statistics:

Mean values and standard deviation were calculated.

Results and discussion

Test results

Test results 1

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:

cytotoxicity

Remarks:

TA 100 at 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING: In a preliminary experiment employing all five applied tester strains triplicates of controls and single plates with test substance (50, 150, 500, 1500 or 5000 µg/plate) were incubated to determine cytotoxicity in the presence or absence of S9. No cytotoxicity was observed in the pretest.

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 1 (plate incorporation)

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		Cross-linking type	Frameshift type
		TA 100	TA 1535	WP2	TA 98	TA 1537
–	0	135	13	22	19	7
–	312.5	114	10	20	21	7
–	650	118	12	18	22	6
–	1250	105	11	20	18	4
–	2500	104	9	18	21	4
–	5000	90*	8	22	17	4
Positive controls, -S9	Name	AF2	SA	AF2	AF2	9AA
	Concentrations (μg/plate)	0.01	0.5	0.01	0.1	80
	Mean No. of colonies/plate (average of 3)	523	349	156	629	1114
+	0	129	12	24	29	9
+	312.5	124	10	29	34	13
+	650	138	15	29	38	12
+	1250	138	13	28	32	13
+	2500	128	18	29	31	11
+	5000	142	17	29	25	12
Positive controls, +S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	1	2	10	0.5	2
	Mean No. of colonies/plate (average of 3)	1015	205	1510	290	207

 

Table 2: Test results of experiment 2 (plate incorporation)

With or without S9-Mix	Test substance concentration	Mean number of revertant colonies per plate (average of 3 plates)
		Base-pair substitution type		Cross-linking type	Frameshift type
		TA 100	TA 1535	WP2	TA 98	TA 1537
–	0	119	14	28	26	9
–	312.5	113	8	20	26	6
–	650	127	9	29	22	4
–	1250	121	9	19	28	6
–	2500	114	9	21	24	6
–	5000	90*	8	20	24	5
Positive controls, –S9	Name	AF2	SA	AF2	AF2	9AA
	Concentrations (μg/plate)	0.01	0.5	0.01	0.1	80
	Mean No. of colonies/plate (average of 3)	612	450	143	870	1882
+	0	125	13	21	34	15
+	312.5	134	9	22	33	13
+	650	145	14	24	40	14
+	1250	138	10	28	39	12
+	2500	149	12	24	36	13
+	5000	131	16	28	34	14
Positive controls, + S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	1	2	10	0.5	2
	Mean No. of colonies/plate (average of 3)	1106	310	1512	310	300

AF2 = 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide

9AA = 9-aminoacridine

2AA = 2-aminoanthracene

SA = sodium azide

* = Inhibition was observed against growth of the bacteria.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative