1-Naphthalenesulfonic acid, 6-diazo-5,6-dihydro-5-oxo-, ester with phenyl(2,3,4-trihydroxyphenyl)methanone

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:

guideline study with acceptable restrictions

Remarks:

No historical control values given

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1982

Report date:

1982

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Version / remarks:

adopted in 2008

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1-Naphthalenesulfonic acid, 6-diazo-5,6-dihydro-5-oxo-, ester with phenyl(2,3,4-trihydroxyphenyl)methanone

EC Number:

270-931-7

EC Name:

1-Naphthalenesulfonic acid, 6-diazo-5,6-dihydro-5-oxo-, ester with phenyl(2,3,4-trihydroxyphenyl)methanone

Cas Number:

68510-93-0

Molecular formula:

all potential esters of C10HH5O3N2SCl and C13H10O4

IUPAC Name:

3-benzoyl-2-hydroxy-6-({[5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalen-1-yl]sulfonyl}oxy)phenyl 5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalene-1-sulfonate; 4-benzoyl-2,3-bis({[5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalen-1-yl]sulfonyl}oxy)phenyl 5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalene-1-sulfonate; 4-benzoyl-2,3-dihydroxyphenyl 5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalene-1-sulfonate

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature, 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, TA 100 and E. coli WP2

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

S-9 mix of male rat liver

Test concentrations with justification for top dose:

4, 20, 100, 500, 2500, 10000 µg/plate (Range Finding Test)
4, 20, 100, 500, 2500, 5000 µg/plate (Main Test)

Vehicle / solvent:

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

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: 2-aminoanthracene (2AA), N-Methyl-N-nitro-nitrosoguanidine (MNNG)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation) (Range Finding Test and Main Test)

DURATION
- Exposure duration: 48 - 72 h

NUMBER OF REPLICATIONS: triplicates

DETERMINATION OF CYTOTOXICITY
- Method: revertant colony number and inspection of the bacterial background lawn

Evaluation criteria:

no information provided

Statistics:

Mean values were calculated.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

number of revertant colonies decreased to less than half compared to solvent control starting at 2500 μg/plate without S9 mix (100%) and with S9 mix (52 and 94%)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

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 to solvent control starting at 2500 μg/plate without S9 mix (78 and 100%) and at 5000 μg/plate with S9 mix (85%)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

number of revertant colonies decreased to less than half compared to solvent control at 100 μg/plate, 2500 and 5000 μg/plate without S9 mix (53, 80 and 100%) and with S9 mix (60 and 90%)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

number of revertant colonies decreased to less than half compared to solvent control at 500 and 5000 μg/plate without S9 mix (54 and 100%) and starting at 2500 μg/plate with S9 mix (58 and 89%)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

number of revertant colonies decreased to less than half compared to solvent control at 5000 μg/plate with S9 mix (61%)

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:

no cytotoxicity, but tested up to precipitating concentrations

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 in each strain

RANGE-FINDING/SCREENING STUDIES: The Range-finding study (performed in all strains) resulted in toxicity at 10000 μg/plate in all strains with and without metabolic activation. Additionally two further strains showed signs of toxicity at 2500 μg/plate without S9 mix (TA 1535, TA 1537 and TA 1538) and with S9 mix (TA 100, TA 1535 and TA 1538) . In TA 1538 toxicity could be observed furthermore at 4, 100 and 500 μg/plate without S9 mix. In TA 1535 toxicity could be observed at 4 and 500 μg/plate with S9 mix. TA 1537 showed a very low value of reverted colonies in the vehicle control with S9 mix (8).



ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Measurement of cytotoxicity used: [complete, e.g. CBPI or RI in the case of the cytokinesis-block method; RICC, RPD or PI when cytokinesis block is not used]
- Other observations when applicable: [complete, e.g. confluency, apoptosis, necrosis, metaphase counting, frequency of binucleated cells]

Remarks on result:

other: in addition, incomplete bacterial lawn starting at 2500 μg/plate with and without S9 mix

Any other information on results incl. tables

Table 1: Summary of test results (Range Finding Test, Plate Incorporation Method)

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Frameshift type			Base-pair substitution type
		TA98	TA1537	TA1538	TA100	TA1535	WP2 uvrA
–	Solvent control (DMSO)	26	8	27	75	17	32
	4	27	13	9	95	24	28
	20	31	7	15	97	20	30
	100	41	8	9	84	11	23
	500	31	8	7 I, P	105	15	24
	2500	19 P	5 P	0 I, P	56 P	6 P	20 P
	10000	2 I, P	0 I, P	0 I, P	0 I, P	0 I, P	5 I, P
	Positive controls (µg/plate)	2NF (5)	9AA (50)	2NF (5)	SA (1)	SA (1)	MNNG (10)
	Mean (No. of colonies/plate)	674	641	1013	667	322	1867
+	Solvent control (DMSO)	23	8	22	93	17	23
	4	22	37	13	103	8	22
	20	33	9	19	89	11	30
	100	25	10	22	115	14	18
	500	28	13	12	128	8	33
	2500	28 P	9 P	5 P	45 I, P	7 P	26 P
	10000	3 I, P	0 I, P	0 I, P	0 I, P	0 I, P	4 I, P
	Positive controls (µg/plate)	2AA (1)	2AA (1)	2AA (1)	2AA (1)	2AA (1)	2AA (10)
	Mean (No. of colonies/plate)	723	80	915	925	186	193
	Positive controls (µg/plate)	B(a)P (10)	B(a)P (10)	B(a)P (10)	B(a)P (10)	B(a)P (10)	B(a)P (10)
	Mean (No. of colonies/plate)	654	141	244	456	17	58

2AA = 2-aminoanthracene

2NF = 2-nitrofluorene

9AA = 9-aminoacridine

B(a)P = benzo(a)pyrene

MNNG = N-Methyl-N-nitro-nitrosoguanidine

SA = sodium azide

I = incomplete bacterial lawn

P = visible precipitation

Table 2: Summary of test results (Main Test, Plate Incorporation Method)

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Frameshift type			Base-pair substitution type
		TA98	TA1537	TA1538	TA100	TA1535	WP2 uvrA
–	Solvent control (DMSO)	17	15	11.00	116.00	9.00	26.00
	4	17	10	13.00	113.00	9.00	19.00
	20	18	11	15.00	93.00	8.00	20.00
	100	16	7.00	10.00	97.00	7.00	25.00
	500	20	9.00	5.00	72.00	8.00	18.00
	2500	12 I, P	3 I, P	6 I, P	0 I, P	2 I, P	16 P
	5000	9 I, P	0 I, P	0 I, P	0 I, P	0 I, P	18 P
	Positive controls (µg/plate)	2NF (5)	9AA (50)	2NF (5)	SA (1)	SA (1)	MNNG (10)
	Mean (No. of colonies/plate)	674	641	1013	667	322	1867
+	Solvent control (DMSO)	28	10.00	19.00	159.00	13.00	24.00
	4	28	15.00	18.00	161.00	15.00	21.00
	20	26	11.00	17.00	163.00	11.00	18.00
	100	24	11.00	20.00	163.00	7.00	28.00
	500	21	7.00	18.00	185.00	10.00	22.00
	2500	22 I, P	4 I, P	8 I, P	76 I, P	7 I, P	20 P
	5000	11 I, P	1 I, P	2 I, P	9 I, P	2 I, P	14 P
	Positive controls (µg/plate)	2AA (1)	2AA (1)	2AA (1)	2AA (1)	2AA (1)	2AA (10)
	Mean (No. of colonies/plate)	723	80	915	925	186	193
	Positive controls (µg/plate)	B(a)P (10)	B(a)P (10)	B(a)P (10)	B(a)P (10)	B(a)P (10)	B(a)P (10)
	Mean (No. of colonies/plate)	654	141	244	456	17	58

2AA = 2-aminoanthracene

2NF = 2-nitrofluorene

9AA = 9-aminoacridine

B(a)P = benzo(a)pyrene

MNNG = N-Methyl-N-nitro-nitrosoguanidine

SA = sodium azide

I = incomplete bacterial lawn

P = visible precipitation