Reaction mass of 1,8-diazabicyclo[5.4.0]undec-7-ene and 2H-Azepin-2-one, 1-(3-aminopropyl)hexahydro-

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

24556-95-8

IUPAC Name:

24556-95-8

Details on test material:

- Name of test material (as cited in study report): Aminopropylcaprolactam
- Physical state: liquid, yellow
- Homogeneity: homogeneous by visual inspection
- Analytical purity: 54.8% (main component, GC)
- Impurities (identity and concentrations): ca. 10% 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU; CAS 6674-22-2), ca. 6% epsilon-caprolactam (CAS 105-60-2), ca. 12% 3,3'-oxybis-1-propanamine (CAS 2157-24-6), ca. 3.5% 3-aminopropan-1-ol (CAS 156-87-6), water
- Lot/batch No.: K 99/2
- Date of manufacturing: June 13, 1996
- Stability under test conditions: The stability of the TS over the study period has been proven by reanalysis.
- Storage condition of test material: refrigerator

Method

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:

Aroclor 1254-induced rat liver S-9 mix

Test concentrations with justification for top dose:

40, 200, 1000, 5000, 10000 µg/plate (Standard plate test; SPT)
20, 100, 500, 2500, 5000 µg/plate (Preincubation test; PIT)

Vehicle / solvent:

- Vehicle used: water

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Remarks:

W/o S-9: N-methyl-N'-nitro-N-nitroso-guanidine (TA 100 , TA 1535), 4-nitro-o-phenylenediamine (TA 98), 9-aminoacridinie chloride monohydrate (TA 1537), N-ethyl-N'-nitro-N-nitroso-guanidine E. coli WP2 uvrA; W/ S-9: 2-aminoanthracene (all strains)

Details on test system and experimental conditions:

METHOD: standard plate test (SPT) and preincubation test (PIT)

DURATION:
- Exposure duration: 48 - 72 hours

NUMBER OF REPLICATIONS:
2 experiments (No. 1: SPT; No. 2: PIT), 3 plates per dose or per control

DETERMINATION OF CYTOTOXICITY
- Method: his- or trp- backgroung growth, decrease in the number of his+ or trp+ revertants, reduction in the titer

Evaluation criteria:

In general, a substance to be characterized as positive in the bacterial tests has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results.

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:

depending on the strain at doses ≥ 5000 µg/plate (SPT) or ≥ 2500 µg/plate (PIT)

Vehicle controls validity:

valid

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:

cytotoxicity

Remarks:

depending on the strain at doses ≥ 5000 µg/plate (SPT) or ≥ 2500 µg/plate (PIT)

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No test substance precipitation was found.

ADDITIONAL INFORMATION ON CYTOTOXICITY: A bacteriotoxic effect was observed depending on the strain and test conditions at doses ≥ 5,000 µg/plate (SPT) or ≥ 2,500 µg/plate (PIT).

Any other information on results incl. tables

Frequency of mutants (means) in the Standard plate test:

Test concentration (µg/plate)	TA98		TA100		TA1535		TA1537		E. coli WP2 uvrA
	- S-9	+ S-9	- S-9	+ S-9	- S-9	+ S-9	- S-9	+ S-9	- S-9	+ S-9
Vehicle control	25	39	119	126	18	19	8	9	33	34
40	25	35	124	116	17	13	8	8	38	34
200	21	34	129	139	12	12	9	9	34	28
1000	18	32	114	112	12	11	8	8	36	34
5000	19	21	109	100	6	10	6	6	27	26
10000	12B	4B	72B	69B	4B	7B	4B	4B	20	20
Positive control	692	864	1214	1531	1096	1033	777	224	560	262

B: reduced background growth

 

Frequency of mutants (means) in the Preincubation test:

Test concentration (µg/plate)	TA98		TA100		TA1535		TA1537		E. coli WP2 uvrA
	- S-9	+ S-9	- S-9	+ S-9	- S-9	+ S-9	- S-9	+ S-9	- S-9	+ S-9
Vehicle control	29	48	128	138	21	21	13	13	35	45
20	34	49	119	141	16	14	10	16	39	37
100	33	42	129	133	17	16	9	10	39	36
500	36	57	127	140	17	18	7	13	37	45
2500	17B	52	88B	135	17	15	7B	10	26	30
5000	19B	19B	73B	114B	12B	17B	7B	9B	14B	20B
Positive control	1466	925	1132	1097	1200	124	457	112	530	204

B: reduced background growth

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

According to the results of the present study, the test substance Aminopropylcaprolactam is not mutagenic in the Ames test and in the Escherichia coli - reverse mutation assay under the experimental conditions chosen here.

Executive summary:

The substance Aminopropylcaprolactam was tested for mutagenicity in the Ames test and in the E. coli -reverse mutation assay both in the standard plate test (SPT) and in the preincubation test (PIT) with and without the addition of a metabolizing system obtained from rat liver (S-9 mix) using the Salmonella strains TA 1535, TA 100, TA 1537, TA 98 and Escherichia coli WP2 uvrA. Test concentrations in the range of 40 µg - 10000 µg/plate and 20 µg - 5000 µg/plate were applied in the SPT and PIT, respectively.

An increase in the number of his⁺ or trp⁺ revertants was not observed both in the SPT and in the PIT either without S-9 mix or after the addition of a metabolizing system. A bacteriotoxic effect was observed depending on the strain and test conditions at doses ≥ 5000 µg/plate (SPT) or ≥ 2,500 µg/plate (PIT). No precipitation of the test substance was found.

Conclusion: According to the results of the present study, the test substance Aminopropylcaprolactam is not mutagenic in the Ames test and in the Escherichia coli - reverse mutation assay under the experimental conditions chosen here.