4-(2-phenylpropan-2-yl)phenyl 3-diazo-4-oxo-3,4-dihydronaphthalene-1-sulfonate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

4 valid concentrations, 2-Aminoanthracene as sole indicator of the efficacy of S9-mix, no information on historical data, different positive controls than in TG OECD 471, limited information on material and methods, no cytotoxic evaluation provided

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1981

Report date:

1981

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted 1997

Deviations:

yes

Remarks:

4 valid concentrations, 2-Aminoanthracene as sole indicator of the efficacy of S9-mix, no information on historical data, different positive controls than in TG OECD 471, limited information on material and methods, no cytotoxic evaluation provided

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-(2-phenylpropan-2-yl)phenyl 4-oxo-3-(λ⁵-diazynylidene)-3,4-dihydronaphthalene-1-sulfonate

Cas Number:

23121-00-8

Molecular formula:

C25H20N2O4S

IUPAC Name:

4-(2-phenylpropan-2-yl)phenyl 4-oxo-3-(λ⁵-diazynylidene)-3,4-dihydronaphthalene-1-sulfonate

Test material form:

solid

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: protected from light

Method

Target gene:

his operon, 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:

S. typhimurium TA 1538

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A

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:

4, 20, 100, 500, 2500 and 10000 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

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

other: 2-aminoanthracene (2AA), methylhydrazone derivative (MHD), streptocotocin (SC), ethyl-N-nitro-N'-nitrosoguanidine (ENNG)

Remarks:

no information which solvent was used for positive controls and test substance

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: quadruplicates

DETERMINATION OF CYTOTOXICITY
- Method: revertant colony number

Evaluation criteria:

A chemical is considered to have a positive response, if it shows an increase in the number of induced revertants of more than two times compared to that of the solvent control.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Number of revertant colonies decreased to less than half compared with solvent control at 500 μg/plate (56%) without metabolic activation. Tested up to precipitating concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

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 applicable

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

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 applicable

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

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 applicable

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1538

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 applicable

Positive controls validity:

valid

Test results 6

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Number of revertant colonies decreased to less than half compared with solvent control starting at 2500 μg/plate (65 and 70%) with metabolic activation.

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: yes, starting at 2500 μg/plate with and without S9 mix in the plates of all tested strains

Any other information on results incl. tables

Table 1: Summary of test results (Plate Incorporation Method)

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 4 plates)
		Frameshift type			Base-pair substitution type
		TA 98	TA 1537	TA 1538	TA 100	TA 1535	WP2 uvr A
–	Solvent control (DMSO)	28.00	8.25	17.50	159.00	28.75	30.75
	4	28.25	6.75	19.00	145.75	25.75	46.25
	20	33.50	6.75	13.50	108.75	22.75	49.00
	100	28.75	6.50	11.75	84.25	20.50	44.25
	500	31.75	7.00	12.25	103.50	12.50	38.00
	2500	33.50 P	6.50 P	12.75 P	121.75 P	20.00 P	34.25 P
	10000	36.25 P	6.50 P	14.50 P	109.25 P	19.50 P	20.50 P
	Positive controls (µg/plate)	MHD (5)	9AA (100)	MHD (5)	MHD (5)	SC (5)	ENNG (1)
	No. of colonies/plate (plate 1 and 2)	3360 4160	2100 2180	2360 2770	>5000 >5000	>5000 >5000	175 202
	Positive controls (µg/plate)	2AA (0.5)	2AA (1)	2AA (0.5)	2AA (0.5)	2AA (1)	2AA (10)
	No. of colonies/plate (plate 1 and 2)	33 41	8 9	13 15	112 121	23 24	20 46
+	Solvent control (DMSO)	39.75	12.00	23.00	167.75	27.75	58.50
	4	34.25	11.75	23.00	157.00	29.00	35.25
	20	37.25	10.50	24.75	163.75	27.00	36.00
	100	31.25	10.75	24.75	146.00	30.00	40.00
	500	32.25	9.75	19.75	145.25	29.50	35.50
	2500	36.25 P	8.25 P	19.00 P	121.25 P	27.00 P	20.50 P
	10000	30.75 P	10.00 P	15.25 P	119.00 P	25.00 P	17.75 P
	Positive controls (µg/plate)	2AA (0.5)	2AA (1)	2AA (0.5)	2AA (0.5)	2AA (1)	2AA (10)
	No. of colonies/plate (plate 1 and 2)	705 815	164 176	780 785	775 785	143 154	1890 2530

2AA = 2-aminoanthracene

9AA = 9-aminoacridine

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

MHD = methylhydrazone derivative

SC = streptocotocin

P = precipitation observed