Carboxylic acids, di-, C5-9

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Remarks:

review article with limited information

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

publication

Title:

THE RESULTS OF MICROBIAL MUTATION TEST FOR FORTY-THREE INDUSTRIAL CHEMICALS

Author:

Shimizu, H. et al.

Year:

1985

Bibliographic source:

Jpn. J. Ind. Health. 27:400 - 419

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted July 1997

Deviations:

yes

Remarks:

Positive control substance for TA 1535 (without S9-mix) differs from the guideline. S9 was prepared from rats treated with polychlorinated biphenyl (KC 500).

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Adipic acid

EC Number:

204-673-3

EC Name:

Adipic acid

Cas Number:

124-04-9

Molecular formula:

C6H10O4

IUPAC Name:

adipic acid

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

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the liver of male rats treated with polychlorinated biphenyl (KC 500) at a dose of 500 mg/kg bw

Test concentrations with justification for top dose:

all strains: 1, 5, 10, 50, 100, 500, 1000 and 5000 µg/plate with and without metabolic activation

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

details are given below (see additional information on materials and methods)

Details on test system and experimental conditions:

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: The test was performed in duplicate.

DETERMINATION OF CYTOTOXICITY
The background bacterial lawn was checked by a dissection.

Statistics:

For the test item, mean values of the duplicates are shown whereas mean values and standard deviations are given for the positive control substances and vehicle controls.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

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

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 6

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Any other information on results incl. tables

Table 1. Test results

With or without S9-Mix	Test item concentration (µg/plate)	Mean number of revertant colonies/plate ± standard deviation
		Base-pair substitution type			Frameshift type
		TA100	TA1535	WP2 uvrA	TA98	TA1537	TA1538
-	vehicle control (DMSO)	150 ± 16.8	30 ± 5.6	30 ± 9.7	32 ± 7.7	18 ± 8.6	22 ± 7.3
-	1	152	40	58	27	11	15
	5	148	43	46	30	9	18
-	10	151	32	46	26	6	15
-	50	155	36	50	23	9	15
-	100	179	42	40	29	6	20
-	500	154	35	44	31	10	16
-	1000	150	37	43	27	7	18
-	5000	150	34	42	35	9	15
-	Positive controls	AF-2	ENNG	AF-2	AF-2	9AA	4NQO
	Mean No. of colonies/plate ± SD	501 ± 84.7	1101 ± 683.1	1082 ± 293.7	278 ± 64.8	889 ± 275.7	270 ± 66.2
+	vehicle control (DMSO)	154 ± 17.5	15 ± 7.1	34 ± 10.9	42 ± 10.1	22 ± 6.3	28 ± 5.7
+	1	179	15	58	33	15	27
+	5	161	17	56	37	20	24
+	10	174	19	52	46	18	26
+	50	162	19	40	35	15	23
+	100	156	23	49	42	16	30
+	500	158	14	55	39	18	27
+	1000	184	20	50	46	15	23
+	5000	177	19	61	31	17	30
+	Positive controls	B(a)P	2AA	2AA	B(a)P	B(a)P	B(a)P
	Mean No. of colonies/plate ± SD	1084 ± 236.3	440 ± 198.6	359 ± 127.0	809 ± 108.4	313 ± 41.6	354 ± 89.4

AF-2 = 2-(2 -furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide)

ENNG = N-ethyl-N´-nitro-N-nitrosoguanidine

9AA = 9-aminoacridine

4NQO = 4-nitroquinoline-1-oxide

B(a)P = benzo(a)pyrene

2AA = 2-aminoanthracene

SD = standard deviation

Applicant's summary and conclusion

Conclusions:

The test item, adipic acid (CAS 124-04-9), was tested for bacterial mutagenicity according to OECD 471, with S. typhimurium strains TA 1535, 1537, 98, 100 and 1538 as well as E. coli WP2 uvr A at concentration levels up to 5000 µg/plate with and without metabolic activation. Based on the results of the conducted study the test item did not exhibit mutagenic properties in bacterial cells.