2,3,4-trihydroxybenzophenone

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 Feb - 15 Mar 1985

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted Jul 1997

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,3,4-trihydroxybenzophenone

EC Number:

214-540-1

EC Name:

2,3,4-trihydroxybenzophenone

Cas Number:

1143-72-2

Molecular formula:

C13H10O4

IUPAC Name:

4-benzoylbenzene-1,2,3-triol

Test material form:

solid

Method

Target gene:

his operon and 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:

Pre-experiment: 4, 20, 100, 500, 2500 and 10000 µg/plate with and without metabolic activation
Main Experiment:
Experiment I:
The pre-experiment served as Experiment I.
Experiment II:
4, 20, 100, 500, 2500 and 5000 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

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

benzo(a)pyrene

other: -S9: 2-nitrofluorene (5 µg/plate) for TA1538 and TA98; N-Methyl-N'-nitro-N-nitrosoguanidine (2.5 µg/plate) for WP2uvrA; +S9: 2-aminoanthracene (0.5, 1 or 10 µg/plate) for all strains

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 - 72 h

NUMBER OF REPLICATIONS: 2 and 3 replications each in 2 independent experiments (Experiment 1 and 2), respectively.

DETERMINATION OF CYTOTOXICITY
- Method: A reduced rate of spontaneously occuring colonies and visible thinning of the bacterial lawn were used as indicator for toxicity.

OTHER EXAMINATIONS:
Sterility check: A sterility check was performed with agar plates only containing the test substance with and without S9 mix.

Statistics:

Mean values were calculated

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:

Experiment 1: ≥ 2500 µg/plate without metabolic activation and ≥ 500 µg/plate with metabolic activation; Experiment 2: ≥ 2500 µg/plate with and without metabolic activation

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:

cytotoxicity

Remarks:

Experiment 1 and 2: ≥ 2500 µg/plate with and without metabolic activation

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:

cytotoxicity

Remarks:

Experiment 1 and 2: ≥ 2500 µg/plate with and without metabolic activation

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:

cytotoxicity

Remarks:

Experiment 1 and 2: ≥ 2500 µg/plate with and without metabolic activation

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:

cytotoxicity

Remarks:

Experiment 1 and 2: ≥ 2500 µg/plate with and without metabolic activation

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:

Experiment 1 and 2: ≥ 2500 µg/plate with and without 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: Precipiation of the test substance on the plates was observed in the highest concentration of 10000 µg/plate with and without metabolic activation.

RANGE-FINDING/SCREENING STUDIES: The pre-experiment served as Experiment I (all strains were tested in the pre-experiment).

HISTORICAL CONTROL DATA
- Positive historical control data: The historical control data of the positive controls were not provided in the study report, but the control plates with the background control gave the expected number of colonies.
- Negative (solvent/vehicle) historical control data: No historical control data of the vehicle control was provided in the study report, but the control plates with the positive controls gave the expected number of colonies.

OTHER EXAMINATIONS:
Sterility of S-9 Mix and the test compound was indicated by the absence of contamination on the test material and S-9 Mix sterility check plates.

Any other information on results incl. tables

Table 1: Summary of Experiment 1

EXPERIMENT 1
S9-Mix	Without
Test item (µg/plate)	TA100	TA1535	TA1537	TA1538	TA98	WP2uvrA
0	144	19	10	12	27	16
4	164	18	8	10	19	27
20	161	14	11	13	24	22
100	175	14	8	13	27	22
500	160	11	7	14	23	17
2500	9 (ibl)	ibl	nbl	0 ibl	0 (ibl)	4 (ibl)
10000	0 P (nbl)	0 P (nbl)	0 P (nbl)	0 P (nbl)	0 P (nbl)	0 P (nbl)
Positive controls	SA	SA	9-AA	2-NF	2-NF	MNNG
Concentrations (μg/plate)	1	1	50	5	5	2.5
Mean No. of colonies/plate	560	384	95	786	450	221
S9-Mix	With
Test item (µg/plate)	TA100	TA1535	TA1537	TA1538	TA98	WP2uvrA
0	145	15	12	16	27	25
4	174	13	11	13	31	23
20	185	13	9	18	31	19
100	196	12	15	17	35	25
500	130 (ibl)	7	11	17	30	19
2500	23 (ibl)	2 (ibl)	0 (ibl)	0 (ibl)	1 (ibl)	4
10000	0 P (nbl)	0 P (nbl)	0 P (ibl)	0 P (nbl)	0 P (nbl)	0 P (nbl)
Positive control	2-AA	2-AA	2-AA	2-AA	2-AA	2-AA
Concentrations (μg/plate)	0.5	1	1	0.5	0.5	10
Mean No. of colonies/plate	680	156	109	538	545	272
Positive control	B(a)P	B(a)P	B(a)P	B(a)P	B(a)P	B(a)P
Concentrations (μg/plate)	10	10	10	10	10	10
Mean No. of colonies/plate	449	29	163	269	704	57

ibl: incomplete background lawn in all plates

(ibl): incomplete background lawn in individual plates

nbl: no background lawn

(nbl): no background lawn in individual plates

P: precipitation

SA: sodium azide

9-AA: 9 -aminoacridine

2-NF: 2 -nitrofluorene

MNNG: N-methyl-N'-nitro-N-nitrosoguanidine

2-AA: 2 -aminoanthracen

B(a)P: benzo(a)pyrene

Table 2: Summary of Experiment 2

EXPERIMENT 2
S9-Mix	Without
Test item (µg/plate)	TA100	TA1535	TA1537	TA1538	TA98	WP2uvrA
0	134	21	13	16	27	20
4	151	18	14	17	31	23
20	160	22	11	8	26	18
100	199	20	11	14	25	19
500	164	17	15	17	27	20
2500	0 (nbl)	nbl	0 (ibl)	0 (ibl)	1 (ibl)	1 (ibl)
5000	0 (nbl)	0 (nbl)	0 (nbl)	0 (nbl)	0 (nbl)	0 (nbl)
Positive controls	SA	SA	9-AA	2-NF	2-NF	MNNG
Concentrations (μg/plate)	1	1	50	5	5	2.5
Mean No. of colonies/plate	601	413	132	192	627	593
S9-Mix	With
Test item (µg/plate)	TA100	TA1535	TA1537	TA1538	TA98	WP2uvrA
0	143	17	12	13	27	19
4	157	9	11	17	28	16
20	191	12	15	17	35	21
100	193	11	15	17	34	18
500	131 (ibl)	11	16	13	36	19
2500	0 (ibl)	1 (ibl)	0 (ibl)	2 (ibl)	3 (ibl)	7 (ibl)
5000	0 (nbl)	0 (nbl)	0 (nbl)	0 (nbl)	0 (nbl)	0 (nbl)
Positive control	2-AA	2-AA	2-AA	2-AA	2-AA	2-AA
Concentrations (μg/plate)	0.5	1	1	0.5	0.5	10
Mean No. of colonies/plate	2862	246	463	2520	2740	324
Positive control	B(a)P	B(a)P	B(a)P	B(a)P	B(a)P	B(a)P
Concentrations (μg/plate)	10	10	10	10	10	10
Mean No. of colonies/plate	635	30	145	245	734	70

ibl: incomplete background lawn in all plates

(ibl): incomplete background lawn in individual plates

nbl: no background lawn

(nbl): no background lawn in individual plates

P: precipitation

SA: sodium azide

9-AA: 9 -aminoacridine

2-NF: 2 -nitrofluorene

MNNG: N-methyl-N'-nitro-N-nitrosoguanidine

2-AA: 2 -aminoanthracen

B(a)P: benzo(a)pyrene