Propazine

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:

from 21 October 1986 to 14 November 1986

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

the statistical analysis has not been carried out

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: AMES, B.N., F.D. LEE and W.E. DURSTON (1973), An Improved Bacterial Test System for the Detection and Classification of Mutagens and Carcinogens. Proc. Natl. Acad. Sci. USA 70, 782786.

Deviations:

not specified

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: AMES, B.N.,W.E. DURSTON, E. YAMASAKI and F.D. LEE (1973), Carcinogens are Mutagens: A Simple Test System Combining Liver Homogenates for Activation and Bacteria for Detection. Proc. Natl. Acad. Sci. USA 70, 22812285.

Deviations:

not specified

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: AMES, B..N., J. McCANN, and E: YAMASAKI (1975), Methods for Detecting Carcinogens and Mutagens with the Salmonella/Mammalian Microsome Mutagenicity Test. Mutation Res. 31, 347364.

Deviations:

not specified

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Propazine

EC Number:

205-359-9

EC Name:

Propazine

Cas Number:

139-40-2

Molecular formula:

C9H16ClN5

IUPAC Name:

6-chloro-N2,N4-bis(propan-2-yl)-1,3,5-triazine-2,4-diamine

Details on test material:

Test material: propazine
Batch No. op. 909005
Purity: 100%
Stability: ensured by the sponsor
Validity (expiration date): ensured by the sponsor

Method

Target gene:

The test substance was tested for mutagenic effects on histidine-auxotrophic mutants of Salmonella typhimurium.

Species / strain

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

not applicable

Additional strain / cell type characteristics:

other: histidine-auxotrophic

Metabolic activation:

with and without

Metabolic activation system:

rat liver microsomes and co-factors

Test concentrations with justification for top dose:

0.008 - 5000 µg/0.1 ml, range in the toxicity test
20 - 5000 µg/0.1 ml, range in the mutagenicity test.

Vehicle / solvent:

Dimethylsulfoxide (DMSO)

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

DMSO

Negative solvent / vehicle controls:

yes

Remarks:

DMDO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: daunorubi-cin-HCl (DAUNOBLASTIN, Farmitalia, Montedison Farmaceutica GmbH, Freiburg, i., Br., Germany) 5 and 10 µg/0.1 ml phosphate buffer

Remarks:

for strain TA 98

Controls 2

Untreated negative controls:

yes

Remarks:

DMSO

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

for strain TA 100

Migrated to IUCLID6: (Fluka, Buchs, Switzerland), 0.125 and 0.25 µg/0.1 ml phosphate buffer

Controls 3

Untreated negative controls:

yes

Remarks:

DMSO

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

for TA 1535

Migrated to IUCLID6: (Fluka, Buchs, Switzerland), 2.5 and 5 µg/0.1 ml bidistilled water

Controls 4

Untreated negative controls:

yes

Remarks:

DMSO

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 9(5) -aminoacridine hydrochloride monohydrate (Fluka, buchs, Switzerland), 50 and 100 µg/0.1 ml dimethylsulfoxide

Remarks:

for TA 1537

Details on test system and experimental conditions:

A preliminary toxicity test was carried out with strain TA 100 without activation with the concentrations ranging from 0.08 to 5000 µg/0l ml. The protocol used was the same as in the mutagenicity test. Accordingly, the concentration of 5000 µg/0.l ml was used as the highest in the mutagenicity test and the tests were performed with the following concentrations of the trial substance without and with microsomal activation: 20, 78, 313, 1250 and 5000 µg/0.1 ml. The substance is dissolved in dimethylsulfoxide (Siegfried, Zofingen, Switzerland). The solution is prepared by treating with ultrasound (Bransonic 220 water bath, 120 W) for 30 minutes at room temperature and sterilizd by filtration through a 0.22 µm filter. Dimethylsulfoxide alone is used for the negative controls. Each Petri dish contains:
1) approx. 20 ml of minimum agar (Agar (Difco Laboratories, Detroit, Michigan, U.S.A.), plus salts (VogelBonner Medium E) and glucose (Fluka, Buchs, Switzerland))
2) 0.1 ml of a solution of the test substance or the vehicle and 0.1 ml of a bacterial culture (in nutrient broth, Difco Laboratories, Detroit, Michigan, U.S.A., 0.8% plus 0.5% NaCl, or nutrient broth No. 2, Oxoid Ltd., Basingstoke, Hants., England, 2.5%) in 2.0 ml of soft agar.
The soft agar is composed of: 100 ml of 0.6% agar solution with 0.6% NaCl. and 10 ml of a solution of 1histidine, 0.5 mM (Fluka, Buchs, Switzerland or Merck, Darmstadt, Germany) and +biotin, 0.5 mM (Sigma, St. Louis, MO., U.S.A.).
In the experiments in which the substance is metabolically activated, 0.5 ml of an activation mixture is added also.
1 ml activation mixture contains: 0.3 ml S9 fraction of liver from rats (Tif:RAIf(SPF)) induced with Aroclor 1254 (Analabs.,Inc., North Haven, Connecticut, U.S.A.) and 0.7 ml of a solution of co-factors.
In the experiments without and with the addition of microsomal activation mixture three Petri dishes are prepared per strain and per group (i.e. per concentration or per control group).
The plates are incubated for about 48 hours at 37 ± 1.5 °C in darkness.

Evaluation criteria:

Criteria for a positive response:
The test substance is considered to be positive in this test system if one or both of the following conditions are met:
a reproducible doubling of the mean number of revertants per plate above that of the negative control at any concentration level for one or more of the following strains: TA 98, TA 1535 and TA 1537,
a reproducible increase of the mean number of revertants per plate for any concentration above that of the negative control by a factor of 1.5 for strain TA 100.
Generally a concentration-related effect should be demonstrable.
Assay acceptance criteria:
A test is considered acceptable if the mean colony counts of the control values of all strains are within the acceptable range derived from experience for mean colony counts ( mean of three values) of spontaneous revertants and if the results of the positive controls meet the criteria for a positive response.
In either case the final decision has to be based on scientific judgement.

Statistics:

see above

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:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

In the experiments performed without and with microsomal activation, comparison of the number of back-mutant colonies in the controls an the cultures treated with the various concentrations of the test substance, revealed no marked deviations.
No evidence of the induction of point mutations by the test substance or by the metabolites of the substance formed as a result of microsomal activation was detectable in the strains of S. typhimurium used in these experiments.

At the concentrations of 1250 and 5000 µg/0.l ml the substance precipitated in soft agar.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative