Reaction mass of sodium 4-chloro-5-alkylbenzotriazolide and sodium 5-chloro-4-alkylbenzotriazolide and sodium 4-chloro-7-alkylbenzotriazolide and sodium 5-chloro-6-alkylbenzotriazolide

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1998

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: US EPA (1997) Toxic Substances Control Act Test Guidelines ; Title 40 CFR Part 799

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

See attachment

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Test animals

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

Male weight: 28 - 30 g
Female weight: 22-24 g

Humidity: 48-60 %
Temperature: 20-24°C

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

1% methylcellulose

Duration of treatment / exposure:

24 hours with positive control
48 hours with negative control

Doses / concentrations

Remarks:

Doses / Concentrations:
Intraperitoneal injection
Basis:
nominal conc.
60, 120, 240 mg/kg BW

No. of animals per sex per dose:

Male: 0 mg/kg; No. of animals: 5; Sacrifice time: 24 hours
Male: 60 mg/kg; No. of animals: 5; Sacrifice time: 24 hours
Male: 120 mg/kg; No. of animals: 5; Sacrifice time: 24 hours
Male: 240 mg/kg; No. of animals: 5; Sacrifice time: 24 hours
Male: 0 mg/kg; No. of animals: 5; Sacrifice time: 48 hours
Male: 240 mg/kg; No. of animals: 5; Sacrifice time: 48 hours
Female: 0 mg/kg; No. of animals: 5; Sacrifice times: 24 hours
Female: 60 mg/kg; No. of animals: 5; Sacrifice times: 24 hours
Female: 120 mg/kg; No. of animals: 5; Sacrifice times: 24 hours
Female: 240 mg/kg; No. of animals: 5; Sacrifice times: 24 hours
Female: 0 mg/kg; No. of animals: 5; Sacrifice times: 48 hours
Female: 240 mg/kg; No. of animals: 5; Sacrifice times: 48 hours

Positive control(s):

mitomycin C
- Route of administration: Orally by intragastric gavage
- Doses / concentrations: 12mg/kg bw

Examinations

Tissues and cell types examined:

Femur bone marrow: proximal epiphysis

Evaluation criteria:

Micronuclei are identified by the following criteria:
- Large enough to discern morphological characteristics
- Should possess a generally rounded shape with a clearly defined outline
- Should be deeply stained and similar in colour to the nuclei of other cells - not black
- Should lie in the same focal plane as the cell
- Lack internal structure, ie they are pyktonic
- There should be no micronucleous-like debris in the area surrounding the cell

Statistics:

Clinical signs and mortalities
Micronucleated immature erythocyte counts (mie)
Micronucleated mature erythocytes (mme)
Proportion of immature erythocytes (%ie/ie+me)

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Doses producing toxicity: The results of a preliminary test indicated that the maximum dose level tolerated was 240 mg/kg. In preliminary tests, dose levels of 320 mg/kg or more produced mortality. There were no mortalities in the main test. Clinical signs were confined to the intermediate and high dose group (both sexes) and consisted of lethargy, loss of righting reflex, splayed limbs and waddling (intermediate group only). Recovery was complete in all animals by 3 hours after dosing. No adverse clinical signs were obtained for the vehicle control or positive control treated animals over the duration of the test. Observations: The test substance did not cause any statistically significant increases in the number of micronucleated immature or mature erythrocytes at either sampling time. The test substance did not cause any significant decreases in the proportion of immature erythrocytes. The positive control caused large, highly significant increases (p<0.001) in the frequency of micronucleated immature erythrocytes.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
Since the test substance did not cause any significant increase in the incidence of micronucleated immature erythocytes or any substantial decrease in the proportion of immature erythocytes, it is conclued that Cl-TTA Solid did not show any evidence of causing chromosome damage or bone marrow cell toxicity when administered by intraperitoneal injection in this in vivo test procedure.