Synthetic resin, a reaction product of 4,4'-bis(N-maleimido)diphenylmethane with 3-aminobenzoic acid hydrazide

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

- study according to OECD guideline 471, GLP, Salmonella typhimurium TA 1535, TA 1537, TA 1538 TA 98 and TA 100 strains exposed to 9.8, 20, 39, 78, 156, 312 µg Compimide 183/plate for 48 h, non-mutagenic

- study according to Abbreviated Japanese MOL/MHW/MITI Metaphase Analysis In CHO Cells In Vitro, similar to OECD guideline 473, Chinese hamster Ovary (CHO) were exposed to 2.5, 5, 10 and 20 µg MDAB/mL for 24h without metabolic activation and 6h with metabolic activation, clastogen

- study according to OECD guideline 471, GLP, Salmonella typhimurium and Escherichia coli strains were exposed to 1, 3, 10, 33, 100, 333, 1000, 3330 and 5000 µg MDAB/plate for 48 h, non-mutagenic, read-across

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1985-03-12 to 1985-04-05

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1981

Deviations:

no

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

other: S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

9.8, 20, 39, 78, 156, 312 µ/plate, the test concentrations were chosen based on bacterial toxicity measured in a preliminary test

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Dimethyl formamide (DMF)
- Justification for choice of solvent/vehicle: Compimide 183 has been shown to be stable in DMF for at least 24h.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

other: 2-Aminofluorene, Neutral red

Details on test system and experimental conditions:

METHOD OF APPLICATION: not specified

DURATION
- Exposure duration: 2 days

NUMBER OF REPLICATIONS: 3 each in two independent experiments

Evaluation criteria:

not specified

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Table 1: In vitro semi-quantitative assays for mutagenicity, using Salmonella typhimurium tester strains in the presence and absence of rat liver microsomes, first assay

Compound	Amount µg/plate	Microsomes	Revertant colonies per plate (a)
			TA 1535	TA 1537	TA 1538	TA 98	TA 100
Compimide 183	312	+	3b	NR	NR	NR	93b
		-	NT	NT	NT	NT	NT
	156	+	11b	1b	18b	25b	160b
		-	2b	NR	8b	7b	103b
	78	+	22b	9b	22b	32b	134
		-	8b	4b	15b	25b	128b
	39	+	22	9	29	30	146
		-	17b	10b	21	26b	116
	20	+	22	8	26	31	139
		-	21b	9	21	22	114
	9.8	+	NT	NT	NT	NT	NT
		-	18	10	21	28	127
Cyclophosamide	250	+	> 500 c	NT	NT	NT	NT
		-	161	NT	NT	NT	NT
Neutral red	25	+	NT	> 500 c	NT	NT	NT
		-	NT	26	NT	NT	NT
2-Aminofluorene	50	+	NT	NT	> 2000 c	> 2000 c	> 1000 c
		-	NT	NT	373	366	164
Dimethyl formamide		+	22	9	22	32	133
		-	20	11	20	26	127

a: Mean value of three replicate plates

b: reduced lawn

c: Visual estimate; checked by auto colony counter

NR: Not recorded due to antibacterial activity

NT: Not tested

> greater than

Table 2: In vitro semi-quantitative assays for mutagenicity, using Salmonella typhimurium tester strains in the presence and absence of rat liver microsomes, second assay

Compound	Amount µg/plate	Microsomes	Revertant colonies per plate (a)
			TA 1535	TA 1537	TA 1538	TA 98	TA 100
Compimide 183	312	+	NR	NR	9b	NR	144b
		-	NT	NT	NT	NT	NT
	156	+	15b	9b	22b	29b	144
		-	NR	NR	10b	17b	131b
	78	+	17b	10	29b	35b	128
		-	13b	7b	13b	25b	147
	39	+	22	10	25	31	152
		-	17b	11b	20	26	152
	20	+	20	12	24	33	170
		-	19b	12	20	25	150
	9.8	+	NT	NT	NT	NT	NT
		-	19	9	19	28	105
Cyclophosamide	250	+	> 500 c	NT	NT	NT	NT
		-	101	NT	NT	NT	NT
Neutral red	25	+	NT	> 500 c	NT	NT	NT
		-	NT	19	NT	NT	NT
2-Aminofluorene	50	+	NT	NT	> 2000 c	> 2000 c	> 1000 c
		-	NT	NT	192	199	181
Dimethyl formamide		+	19	13	25	34	147
		-	19	11	20	29	130

a: Mean value of three replicate plates

b: reduced lawn

c: Visual estimate; checked by auto colony counter

NR: Not recorded due to antibacterial activity

NT: Not tested

> greater than

Conclusions:

In the present study conducted similar to OECD guideline 471, Compimide 183 was tested for mutagenicity in the S. typhimurium tester strains TA 1535, TA 1537, TA 1538, TA 98, TA 100 in the following concentrations: 312, 156, 78, 39, 20, 9.8 µg/plate with and without metabolic activation by S9-liver mix. These concentrations were chosen due to cytotoxicity at 312 µg/plate and above.
There was no increase of revertant colonies up to the highest concentration tested, thus, Compimide 183 is considered to be non-mutagenic.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In a study conducted similar to OECD guideline 471, Compimide 183 was tested for mutagenicity in the s. typhimurium tester strains TA 1535, TA 1537, TA 1538, TA 98, TA 100 in the following concentrations: 312, 156, 78, 39, 20, 9.8 µg/plate with and without metabolic activation by S9-liver mix. These concentrations were chosen due to cytotoxicity at 312 µg/plate and above. There was no increase of revertant colonies up to the highest concentration tested, thus, Compimide 183 is considered to be non-mutagenic.

In a study conducted similar to OECD 473 CHO cells (1E+05 cells/mL) were treated with 2.5, 5.0, 10.0 and 20.0 µg/mL MDAB either 24h without metabolic activation or 6h with metabolic activation. MDAB produced significant, dose-related increases in the frequency of chromosome abberations both in the presence anbd in the absence of a liver enzyme metabolising system. The test item is therefore considered to be clastogenic to CHO cells in vitro.

In a study according to OECD guideline 471, GLP, Salmonella typhimurium and Escherichia coli strains were exposed to 1, 3, 10, 33, 100, 333, 1000, 3330 and 5000 µg MDAB/plate for 48 h, it was concluded that MDAB is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

Justification for classification or non-classification

According to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS) MDAB and Compimide do not need to be classified as mutagenic, but based on the results of a test conducted similar to OECD guideline 473, Compimide 183 needs to be classified as clastogenic according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).