complex reaction mass of Chinese gum rosin post reacted with acrylic acid

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

Study period:

07 July 1998 - 28 July 1998

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): ARAKAWA KE-604
- Physical state: pale yellow blocks
- Analytical purity: no data
- Lot/batch No.: R61132
- Storage condition of test material: room temperature in the dark

Method

Target gene:

genes involved in histidine synthesis

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:

S9-mix

Test concentrations with justification for top dose:

0, 50, 150, 500, 1500, 5000 µg/plate

Vehicle / solvent:

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

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-ethyl-N-nitro-N-nitrosoguanidine; 9-aminoacridine; 4-nitroquinoline-N-oxide; 2-Aminoanthracene; benzo(a)pyrene.

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (direct plate incorporation);

DURATION
- Exposure duration: 48 h at 37ºC

NUMBER OF REPLICATIONS:
Experiment 1: Five concentrations of the test material were assayed in triplicate against each tester strain.
Experiment 2: Methodology as for experiment 1, using fresh bacterial cultures, test material and control solutions.

DETERMINATION OF CYTOTOXICITY
growth of bacterial background lawn.

Evaluation criteria:

The test material should have induced a reproducible, dose-related and statistically (Dunnett´s method of linear regression) significant increase in the revertant count in at least one strain of bacteria. If a greater than 2-fold increase in revertant count is observed in two experiments then this is taken as evidence of a positive response.

Statistics:

Dunnett´s method of linear regression

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:

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

Remarks:

of 5000 µg/plate, at which precipitation occured.

Vehicle controls validity:

valid

Untreated negative 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:

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

Remarks:

of 5000 µg/plate at which precipitation occured

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation:observed at concentrations of 5000 µg/plate

RANGE-FINDING/SCREENING STUDIES:
The material was non-toxic to the strains of bacteria used (TA100 and WP2uvrA-).
The dose range of test material: 0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500, 5000 µg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA:
The number of revertant colonies in the historical vehicle and positive controls were in the range of the concurrent controls.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
The material was non-toxic to the strains of bacteria used up to the limit concentration of 5000 µg/plate as determined by the growth of the background lawn.

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

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			Frameshift type
		TA 100	TA1535	WP2uvrA-	TA98	TA1537
-	0	124	20	27	26	12
-	50	109	20	31	30	9
-	150	114	25	22	27	6
-	500	98	20	24	30	9
-	1500	101	17	22	18	12
-	5000	103 P	17 P	23 P	20 P	6 P
Positive controls - S9	Name	ENNG	ENNG	ENNG	4NQO	9AA
	Concentrations (μg/plate)	3	5	2	0.2	80
	Mean No. of colonies/plate (average of 3)	440	250	683	140	1018
		TA 100	TA1535	WP2uvrA-	TA98	TA1537
+	0	114	17	32	32	15
+	50	104	15	26	33	12
+	150	99	15	35	35	15
+	500	99	14	20	33	11
+	1500	83	12	27	31	9
+	5000	76 P	11 P	19 P	22	6
Positive controls + S9	Name	2AA	2AA	2AA	BP	2AA
	Concentrations (μg/plate)	1	2	10	5	2
	Mean No. of colonies/plate (average of 3)	791	198	706	510	223
		TA 100	TA1535	WP2uvrA-	TA98	TA1537
Concurrent Negative control		109	20	28	36	9

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

4NQO = 4-nitroquinoline-N-oxide

9AA = 9-aminoacridine

2AA = 2-Aminoanthracene

BP = Benzo(a)pyrene

P = Precipitate

 

Table 2: Test Results of Experiment 2

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			Frameshift type
		TA 100	TA1535	WP2uvrA-	TA98	TA1537
-	0	135	31	25	34	17
-	50	119	29	23	32	13
-	150	129	22	20	27	6
-	500	118	20	18	25	7
-	1500	99	23	21	31	6
-	5000	106	21	27	36	11
Positive controls - S9	Name	ENNG	ENNG	ENNG	4NQO	9AA
	Concentrations (μg/plate)	3	5	2	0.2	80
	Mean No. of colonies/plate (average of 3)	631	165	486	136	1001
		TA 100	TA1535	WP2uvrA-	TA98	TA1537
+	0	128	13	26	34	19
+	50	127	18	27	41	14
+	150	123	16	25	34	14
+	500	120	14	30	36	9
+	1500	113	13	26	27	10
+	5000	111	14	27	41	6
Positive controls + S9	Name	2AA	2AA	2AA	BP	2AA
	Concentrations (μg/plate)	1	2	10	5	2
	Mean No. of colonies/plate (average of 3)	959	252	1184	583	492
		TA 100	TA1535	WP2uvrA-	TA98	TA1537
Concurrent Negative control		129	26	25	35	9

 

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

4NQO = 4-nitroquinoline-N-oxide

9AA = 9-aminoacridine

2AA = 2-Aminoanthracene

BP = Benzo(a)pyrene

P = Precipitate

 

Summary:

Salmonella typhimurium strains TA 1535, TA1537, TA98, TA100 and E. coli strain WP2uvrA- were treated with the test substance ARAKAWA KE-604 in DMSO according to OECD Guideline 471 (Ames Test) in two consecutive experiments. The tested dose range was 50 to 5000 µg/plate. The test material caused no visible reduction in the growth of the bacterial lawn at any dose level. A precipitate was observed at 5000 µg/plate, which did not prevent the scoring of revertant colonies.

No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative The test substance was found to be non-mutagenic in bacteria.