2,4,5-trichloroaniline

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: genome mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable with current OECD guideline with acceptable restrictions; The report does not provide information as to the basis of the dose (substance "as provided" or excluding the water content of 24.8 %)

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

The applied dose was higher than 2000 mg/kg bw

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

2,4,5-trichloroaniline

EC Number:

211-254-9

EC Name:

2,4,5-trichloroaniline

Cas Number:

636-30-6

Molecular formula:

C6H4Cl3N

IUPAC Name:

2,4,5-trichloroaniline

Test animals

Species:

mouse

Strain:

other: Bor : NMRI (SPF Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: F Winkelmann, Borchen, Germany
- Age at study initiation: young adult male and virgin females (about 8-12 weeks of age)
- Weight at study initiation: 31- 41g
- Assigned to test groups randomly: yes
- Housing: in groups of 3-5 animals of one sex
- Diet (e.g. ad libitum): yes
- Water (e.g. ad libitum): yes
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-24°C
- Humidity (%): 45- 55
- Air changes (per hr): 10 times/hr
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: Sept. 5, 1988 To: Nov. 15, 1988

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle/solvent used: polyethylene glycol 400 (Lutrol E400)
- Amount of vehicle (gavage): 20 ml/kg

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
2,4,5-Trichloroaniline was disolved in polyethylene glycol 400 and given orally by stomach tube.

Duration of treatment / exposure:

One application of 7500 mg/kg bw test item to each of the three treatment goups and sacrifice at 24, 48 and 72 hours after application.
One group received the vehicle only and was sacrified 24 hours after treatment.
One additional group received 7500 mg/kg bw of the test item and was held as replacement group.

Frequency of treatment:

once

Post exposure period:

24, 48 and 72 hours

Doses / concentrations

Remarks:

Doses / Concentrations:
0 and 7500 mg/kg bw of 2,4,5-Trichloroaniline
Basis:
no data

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control(s):

cyclophosphamide;
- Route of administration: oral by gavage
- Doses / concentrations: 20 mg/kg bw

Examinations

Tissues and cell types examined:

Bone marrow from the femur

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:
The selection of the 2,4,5-Trichloroaniline dose was based on a pilot test (groups of five animals including both males and females; doses of 5000, 7500, 8500 and 10000 mg/kg bw were administered orally.Following symptoms were recorded: apathy, roughed fur, staggering gait, prone position and difficulty in breathing. One of five animals died in the 7500mg/kg bw group, two of five animals of the 8500 mg/kg bw and four of five in the 10000mg/kg bw group. based on these results, 7500 mg/kg bw 2,4,5-Trichloroaniline was chosen for this test,


TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
One application of 7500 mg/kg bw test item to each of the three treatment goups and sacrifice at 24, 48 and 72 hours after application.
One group received the vehicle only and was sacrified 24 hours after treatment.
One additional group received 7500 mg/kg bw of the test item and was held as replacement group.
One positive control group received cyclophosphamide (20 mg/kg bw) and was sacrificed 24 hr after treatment.

DETAILS OF SLIDE PREPARATION:
schmid's method was used to produce the smears. The smears were stained automatically, "destained" with methanol and rinsed with deionized water. They were then left to dry. Thereafter the smears were treated with xylene and taken out singly for covering. With the aid of some covering agent the cover glass was then placed in position.

METHOD OF ANALYSIS:
The coded slides were evaluated using a light microscope at magnification of about 1000. Normally 1000 polychromatic erythrocytes were counted per animal. The incidence of cells with micronuclei was established by scanning the slides in a meander pattern.

Evaluation criteria:

The number of normochromatic erythocytes per 1000 polychromatic was noted. If the ratio for a single animal amounts to distinctly more than 3000 normochromatic erythocytes per 1000 polychromatic, or if this ratio seems to be likely and no other animals in the group show similar effects, then the case may be regarded as pathological and unrelated to treatment, and the animal may be omitted from the evaluation. Conclusions as to relevant treatment-related alteration of that ratio can only be drawn if the ration for the majority of animals in a treatment group is clearly reduced compared to animals in the negative control.
Assessment criteria:
A test was considered positive if there was a relevant and significant increase in the numbers of polychromatic erythrocytes with micronuclei at any of the intervals in comparison to the negative control. A test was considered negative if there was no relevant or significant increase in the rate of micronucleated polychromatic erythrocytes at any time point.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Remarks:

Symptoms: apathy, roughed fur, staggering gait, pallor, prone position, spasm, shivering and difficulty in breathing. Seven of 40 treated animals dies during the conduct of the test due to acute toxicity of 7500 mg/kg 2,4,5- Trichloroaniline.

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

- micronucleated cells /1000 NCE
negative control (24hr): 0.9 ± 1.1
7500 mg/kg bw test item (24 hr): 0.8 ± 0.6
7500 mg/kg bw test item (48 hr): 1.1 ± 0.9
7500 mg/kg bw test item (72 hr): 1.2 ± 1.3
20 mg/kg bw pos. control (24 hr): 1.3 ± 1.3

- micronucleated cells /1000 PCE
negative control (24hr): 1.2 ± 1.1
7500 mg/kg bw test item(24 hr): 1.5 ± 0.7
7500 mg/kg bw test item(48 hr): 2.1 ± 0.7
7500 mg/kg bw test item(72 hr): 2.4 ± 2.8
20 mg/kg bw pos. control (24 hr): 14.7** ± 5.8

- Ratio of PCE/NCE :
negative control (24hr): 1000/1097 ± 389
7500 mg/kg bw test item (24 hr): 1000/2148* ± 1464
7500 mg/kg bw test item (48 hr): 1000/1566 ± 1849
7500 mg/kg bw test item (72 hr): 1000/ 662 ± 272
20 mg/kg bw pos. control (24 hr): 678 ± 125
*P < 0.02 in non-parametric Wilcoxon ranking test
**P < 0.01 in non-parametric Wilcoxon ranking test

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
The number of micronucleated normochromatic erythrocytes did not increase relevantly in any of the groups. In conclusion, it may be stated that no indications of clastogenic effect of 7500mg/kg bw of 2,4,5 -Trochloroaniline per os were found in the micronucleus test in the mouse (i.e. in a somatictest system in vivo).