2'-chloroacetoacetanilide

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:

1990-03-05 to 1990-03-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study documented and performed according to GLP standards and in compliance with OECD Guideline 471.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2'-chloroacetoacetanilide

EC Number:

202-269-1

EC Name:

2'-chloroacetoacetanilide

Cas Number:

93-70-9

Molecular formula:

C10H10ClNO2

IUPAC Name:

2'-chloroacetoacetanilide

Details on test material:

Identity: P0005
Storage: RT
Expiry date: August 13, 1992
Appearance: White powder

Method

Target gene:

Not applicable.

Species / strain

Species / strain / cell type:

other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Details on mammalian cell type (if applicable):

TA 1535 his G46 rfa uvrB
TA 1537 his C3076 rfa uvrB
TA 1538 his D3052 rfa uvrB
TA 98 his D3052 rfa uvrB pKM 101
TA 100 his G46 rfa uvrB pKM 101

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Mixed-function oxidase systems in the rat liver were stimulated following a single i/p injection of Aroclor 1254

Test concentrations with justification for top dose:

Dose range finding test: 5000, 500, 50, 5 microg/plate.
Mutation tests: 5000, 1500, 500, 150, 50 microg/plate.

Vehicle / solvent:

Dimethylsulphoxide

Controls

Untreated negative controls:

yes

Remarks:

dimethylsulphoxide

Negative solvent / vehicle controls:

yes

Remarks:

dimethylsulphoxide

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene, 9-Aminoacridine, N-Ethyl-N'-nitro-N-nitrosoguanidine, 2-Nitrofluorene

Details on test system and experimental conditions:

For use in tests sub-cultures were grown in Nutrient Broth No. 2 (Oxoid) at 37°C for 10 hours. This culture provided approximately 2 x 10x9
organisms per ml which was assessed photometrically.

Preparation of liver homogenate S—9 fraction:
Species: Rat.
Strain: Sprague—Dawley derived.
Source: Bantin and Kingman Ltd., The Field
Station, Grimston, Aldbrough, Hull,
North Humberside, HU11 4QE.
Age range: 7 — 8 weeks.
Weight range: <300 g.
Diet: Labsure Rodent Diet LAD 1.

Stimulation of rat liver enzymes:
Mixed—function oxidase systems in the rat liver were stimulated following a single i/p injection of Aroclor 1254 (diluted in Arachis oil to 200 mg/ml) at a dosage of 500 mg/kg. On the fifth day of induction, following an overnight starvation, the rats were killed and their livers aseptically removed.

Evaluation criteria:

The mean number of revertant colonies for all treatment groups is compared with those obtained for solvent control groups. The mutagenic activity
of a test material is assessed by applying different criteria.
Revertant colonies were counted using a Biotran Automatic Colony Counter. Any toxic effects of the test substance can be detected by a substantial
reduction in revertant colony counts or by the absence of a complete background bacterial lawn.

Statistics:

Not applicable.

Results and discussion

Test results

Species / strain:

other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

In the preliminary toxicity test P0005 was not toxic towards the tester strains.
Therefore, 5000 microg/plate was chosen as the top dose level in the mutation tests.
The mean number of revertant colonies, together with the individual plate counts for P0005 obtained in the first and second mutation test with the
tester strains are shown in the Table below.
No substantial increases in revertant colony numbers of any of the tester strains were observed following treatment with P0005 at any dose level,
either in the presence or absence of S-9 mix.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Strain	Dose level (microg/plate)	Metabolic activation -/+	Mean revertant colony counts
TA 1535	5000 1500  500 150    50 0 Solvent	- / + - / + - / + - / + - / + - / + - / +	7 / 9 7 / 12  8 / 11 9 / 10 10 / 9  8 / 9 7 / 11
TA 1537	5000 1500  500 150    50 0 Solvent	- / + - / + - / + - / + - / + - / + - / +	9 / 7 7 / 7 12 / 10  6 / 6 7 / 9  5 / 7 7 / 6
TA 1538	5000 1500  500  150    50  0 Solvent	- / + - / + - / + - / + - / + - / + - / +	7 / 7  7 / 7  7 / 11 5 / 7  8 / 9 7 / 9 11 / 13
TA 98	5000 1500  500 150 50 0 Solvent	- / + - / + - / + - / + - / + - / + - / +	16 / 11 18 / 18 21 / 17 18 / 19 20 / 17 20 / 17 19 / 18
TA 100	5000 1500  500 150    50 0 Solvent	- / + - / + - / + - / + - / + - / + - / +	61 / 55 68 / 78  69 / 78 81 / 88  69 / 83 72 / 95  81 / 97

- Absence

+ Presence

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative without metabolic activation
negative with metabolic activation

lt is concluded that, when tested at dose levels up to 5000 microg/plate in dimethylsulphoxide, P0005 was not mutagenic in this bacterial test system.

Executive summary:

The genetic toxicity study in vivo (Ames) was performed in year 1990 according to GLP standards and in compliance with OECD Guideline 471.

In the preliminary dose range finding study with dose levels of up to 5000 microg/plate no toxicity was observed. A top dose level of 5000 g/plate was chosen for the subsequent mutation study. Other dose levels used in the mutation assays were: 1500, 500, 150, 50 microg/plate. The concurrent positive control compounds demonstrated the sensitivity of the assay and the metabolising activity of the liver preparations. No evidence of mutagenic activity was seen at any dose level of P0005 in either mutation test.

Therefore, it is concluded that the test item is not mutagenic in this bacterial system.