Suberonitrile

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP study (only TA 98, TA 100, TA 1535 and TA 1537 tested)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(adopted 1983)

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(adopted 1997)

Deviations:

yes

Remarks:

(only 4 strains; only 2-aminoanthracene as positive control with metabolic activation)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Suberonitrile

EC Number:

211-089-2

EC Name:

Suberonitrile

Cas Number:

629-40-3

Molecular formula:

C8H12N2

IUPAC Name:

octanedinitrile

Details on test material:

- Name of test substance (as cited in study report): Korksaeuredinitril
- Analytical purity: 99.18%
- Substance No.: 91/319
- Batch No.: KW 7376 107-5 P 5
- Physical state: yellowish liquid
- Storage: room temperature

Method

Target gene:

His-operon

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254.

Test concentrations with justification for top dose:

20, 100, 500, 2500, 5000 µg/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: (+ S9): 2-aminoanthracene (2AA; all strains); (- S9): N-methyl-N'-nitro-N-nitrosoguanidine (MNNG; TA 100 and TA 1535), 4-nitro-o-phenylendiamine (NOPD; TA 98), 9-aminoacridine chloride monohydrate (AAC; TA 1537)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation): experiment 1

DURATION
- Exposure duration: 48 h (37 °C in the dark)

METHOD OF APPLICATION: preincubation: experiment 2

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h (37 °C in the dark)

NUMBER OF REPLICATIONS: 3 plates per dose per experiment

DETERMINATION OF CYTOTOXICITY
- Method: reduction in the number of revertant colonies; clearing or diminution of the background lawn; degree of survival of treated cultures

Evaluation criteria:

In general, a substance has to fulfill the follwoing requirements to be characterized as positvie in the Ames test:
- doubling of the spontaneous mutation rate
- dose-response relationship
- reproducibility of the results.

Results and discussion

Test results

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 1537 and TA 100 (5000 µg/plate, (-S9); TA 100 (2500 µg/plate, (+S9))

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no precipitation occurred up to a concentration of 5000 µg/plate.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Standard plate test

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type		Frameshift type
		TA 100	TA1535	TA98	TA1537
–	DMSO	133 ± 5	16 ± 6	35 ± 4	12 ± 2
–	20	125 ± 17	16 ± 1	28 ± 2	9 ± 3
–	100	123 ± 15	13 ± 3	20 ± 3	8 ± 1
–	500	139 ± 15	11 ± 2	23 ± 5	7 ± 3
–	2500	102 ± 1	14 ± 2	29 ± 8	9 ± 6
–	5000	83 ± 6	12 ± 2	29 ± 2	5 ± 3
Positive controls, –S9	Name	MNNG	MNNG	NOPD	AAC
	Concentrations (μg/plate)	5	5	10	100
	Mean No. of colonies/plate (average of 3 ± SD)	1187 ± 49	1220 ± 69	890 ± 200	683 ± 137
+	DMSO	142 ± 6	17 ± 3	38 ± 5	15 ± 3
+	20	126 ± 15	18 ± 2	48 ± 4	12 ± 3
+	100	130 ± 12	15 ± 5	42 ± 8	6 ± 1
+	500	95 ± 18	18 ± 4	37 ± 9	9 ± 2
+	2500	77 ± 7	15 ± 4	31 ± 2	12 ± 6
+	5000	96 ± 16	16 ± 3	27 ± 7	9 ± 2
Positive controls, +S9	Name	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	2.5	2.5	2.5
	Mean No. of colonies/plate (average of 3 ± SD)	1393 ± 29	185 ± 69	680 ± 34	106 ± 8

MNNG = N-methyl-N'-nitro-N-nitrosoguanidine

NOPD = 4-nitro-o-phenylenediamine

AAC = 9-aminoacridine

2AA = 2-aminoanthracene

 

Table 2: Preincubation test

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type		Frameshift type
		TA 100	TA1535	TA98	TA1537
–	DMSO	140 ± 9	22 ± 2	27 ± 2	10 ± 4
–	20	139 ± 12	24 ± 5	28 ± 5	12 ± 1
–	100	151 ± 5	22 ± 3	28 ± 6	11 ± 1
–	500	129 ± 16	23 ± 4	27 ± 3	8 ± 1
–	2500	131 ± 15	20 ± 4	27 ± 7	11 ± 3
–	5000	122 ± 6	21 ± 6	28 ± 4	9 ± 1
Positive controls, –S9	Name	MNNG	MNNG	NOPD	AAC
	Concentrations (μg/plate)	5	5	10	100
	Mean No. of colonies/plate (average of 3 ± SD)	915 ± 75	601 ± 67	1247 ± 6	982 ± 227
+	DMSO	130 ± 7	20 ± 3	40 ± 6	13 ± 3
+	20	131 ± 20	23 ± 5	38 ± 4	13 ± 5
+	100	143 ± 20	20 ± 4	35 ± 3	12 ± 3
+	500	120 ± 11	22 ± 4	42 ± 2	13 ± 1
+	2500	103 ± 2	21 ± 3	44 ± 3	12 ± 2
+	5000	120 ± 18	21 ± 1	34 ± 4	9 ± 2
Positive controls, +S9	Name	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	2.5	2.5	2.5	2.5
	Mean No. of colonies/plate (average of 3 ± SD)	877 ± 36	163 ± 20	536 ± 8	149 ± 13

MNNG = N-methyl-N'-nitro-N-nitrosoguanidine

NOPD = 4-nitro-o-phenylenediamine

AAC = 9-aminoacridine

2AA = 2-aminoanthracene

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative