4-chlorophenol

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:

1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions (no analytical purity reported; no E.coli strain tested)

Data source

Reference

Reference Type:

publication

Title:

Salmonella Mutagenicity Test Result for 250 Chemicals.

Author:

Haworth S, Lawlor T, Mortelmans K, et al.

Year:

1983

Bibliographic source:

Environmental Mutagenesis Supplement 1:3 - 142

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no analytical purity reported; no E.coli strain tested

GLP compliance:

no

Remarks:

prior to GLP

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-chlorophenol

EC Number:

203-402-6

EC Name:

4-chlorophenol

Cas Number:

106-48-9

Molecular formula:

C6H5ClO

IUPAC Name:

4-chlorophenol

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

Cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from livers of rats and hamsters treated with Aroclor 1254.

Test concentrations with justification for top dose:

10, 33.3, 100, 333.3, 1000 µg/plate without and with metabolic activation

Vehicle / solvent:

- Vehicle/solvent used: DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Remarks:

+S9: 1.0 µg/plate 2-aminoanthracene (2-AA) for TA 98, TA 100, 2.5 µg/plate for TA 1535 and TA 1537

Positive control substance:

other: -S9: 5 µg/plate 4-Nitro-ophenylenediamine (NOPD) for TA 98, 1 µg/plate Sodium azide (SA) for TA100 and TA 1535 and 50 µg/plate 9-Aminoacridine (9-AA) for TA 1537

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation
- Salmonella strains TA98, TA100, TA 1535 and TA1537 were used in a modification of the preincubation test of Yahagi et al., 1975 (Mutagenicity of carcinogenic azo dyes and their derivatives. Cancer Lett 1:91-96).
- The preincubation procedure was selected because of reports that it was no less sensitive than the plate test, and was more effective than the plate test for various chemicals such as aliphatic nitrosamines, pyrrolizidine alkaloids, and volatile chemicals.

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h at 37 °C

NUMBER OF REPLICATIONS: three replicates each in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: thinning or absence of the bacterial lawn.

Evaluation criteria:

The test substance may be considered positive in this test system if the following criteria are met: For the test substance to be considered mutagenic, a reproducible dose related increase, whether it is twofold over background or not.

Statistics:

Mean values and standard deviation were calculated.

Results and discussion

Test results

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:

cytotoxicity

Remarks:

yes 1000 µg/plate TA 100, +S9

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: The highest concentration analysed was selected on the solubility of the test substance in cell culture medium.

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 the experiment (preincubation)

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type		Frameshift type
		TA 100	TA1535	TA1537	TA98
–	0.0	89 ± 5.5	19 ± 3.2	13 ± 2.1	23 ± 3.4
–	10.0	62 ± 9.8	10 ± 2.3	5 ± 0.6	18 ± 3.3
–	33.3	65 ± 1.0	11 ± 3.3	8 ± 2.0	20 ± 4.1
–	100.0	55 ± 4.3	14 ± 2.0	7 ± 3.5	18 ± 3.4
–	333.3	62 ± 3.0	8 ± 2.6	6 ± 0.6	17 ± 1.5
–	1000.0	47 ± 10.0	9 ± 1.8	8 ± 0.6	3 ± 2.0
Positive controls, –S9	Name	SA	SA	4NQO	9AA
	Mean No. of colonies/plate   (average of 3 ± SD)	494 ± 8.4	484 ± 9.2	697 ± 292.4	615 ± 9.0
+ (rat)	0.0	73 ± 5.2	4 ± 1.0	5 ± 0.3	24 ± 4.4
+ (rat)	10.0	78 ± 2.8	6 ± 0.7	5 ± 0.6	28 ± 1.8
+ (rat)	33.3	74 ± 2.8	5 ± 1.3	5 ± 0.6	30 ± 1.7
+ (rat)	100.0	94 ± 9.0	6 ± 1.5	5 ± 1.7	30 ± 2.8
+ (rat)	333.3	73 ± 3.8	10 ± 1.2	6 ± 1.9	29 ± 5.6
+ (rat)	1000.0	T	9 ± 0.7	1 ± 7	14 ± 1.5
Positive controls, +S9 (rat)	Name	2AA	2AA	2AA	2AA
	Mean No. of colonies/plate (average of 3 ± SD)	877 ± 45.1	191 ± 28.3	244 ± 10.5	956 ± 33.1
+ (hamster)	0.0	97 ± 10.5	15 ±1.8	33 ± 1.8	41 ± 4.5
+ (hamster)	10.0	77 ± 1.5	8 ± 1.9	8 ± 2.8	28 ± 0.7
+ (hamster)	33.3	70 ± 8.1	11 ± 3.5	9 ± 3.0	27 ± 4.5
+ (hamster)	100.0	75 ± 7.7	11 ± 1.7	10 ± 1.3	28 ± 0.9
+ (hamster)	333.3	75 ± 6.7	14 ± 3.5	8 ± 1.5	26 ± 0.3
+ (hamster)	1000.0	42 ± 12.0	8 ± 0.6	7 ± 1.3	28 ± 0.6
Positive controls, +S9 (ha mster)	Name	2AA	2AA	2AA	2AA
	Mean No. of colonies/plate (average of 3 ± SD)	1215 ± 16.4	458 ± 3.3	245 ± 7.5	1397 ± 41.3