Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well documented study according to international accepted guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
Test item: 2,3-DICHLOROBENZOYL NITRILE
CAS No.: 77668-42-9
Batch No.: L36020N
Physical state: Yellow, crystalline powder
Colour: Light-yellow
Purity (GC): 96.48% (based on 2,3-DICHLOROBENZOYL NITRILE content)
Storage 15-30°C

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Species and strain: Crl:(WI)Br rats
Source: TOXI COOP ZRT.
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Number of animals: 3 animals/group
Sex: Female, nulliparous and non pregnant animals
Age of animals: Young adult rat, 8 weeks old in first, second and third step
Body weight range at starting (first step): 155 - 162 g
Body weight range at starting (second step): 170 - 175 g
Body weight range at starting (third step): 166 - 178 g
Body weight range at starting (fourth step): 171 - 172 g
Acclimatization time: 5 days in first step, 6 days in second step, 7 days in third step and 8 days in fourth step
Animal health: Only healthy animals were used for the study

Housing: Group caging (3 animals/cage)
Light: Artificial light, from 6 a.m. to 6 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: 10-15 air exchanges/hour by central air-condition system

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Helianthi annui oleum raffinatum
Details on oral exposure:
A single oral administration - followed by a fourteen-day observation period - was performed by gavage.
Starting dose was selected on the basis of the available information about the test item. The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. Since all female animals died the test was continued at 300 mg/kg bw dose level on further three female rats. All animals died in the second step at 300 mg/kg bw dose level, the test was continued at 50 mg/kg bw dose level on further three female rats. There was no death in third step, so three further female rats were treated with the same (50 mg/kg bw) dose. No animal died in the fourth step, too, so the test was finished.

All doses were formulated in the vehicle. Concentration of formulations was adjusted to maintain a treatment volume of 10 mL/kg bw. The test item was applied in a concentration of 200, 30 and 5 mg/mL. Formulations were prepared just before the administration and stirred continuously during the treatment.

The day before treatment the animals were fasted. The food but not water was withheld overnight. The food was given back 3 hours after the treatment.
Doses:
2000, 300, 50 mg/kg bw
No. of animals per sex per dose:
3 female/dose
Control animals:
no
Details on study design:
The observation period was 14 days. Animals were observed individually after dosing at least once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h after the treatment and twice each day for 14 days thereafter.
The body weights were recorded on day 0 (just before the treatment), on day 7 and on day 15 with a precision of 1 g.
At the end of the observation period rats were sacrificed and necropsy were performed.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
50 - 300 mg/kg bw
Based on:
test mat.
Mortality:
All rats dosed at 2000 mg/kg “2,3-dichlorobenzoyl nitrile” died. All females of group 1 died on the treatment day, 7 min. after the treatment.
Besides, all rats treated with 300 mg/kg dose died on the treatment day. Two animals died 15 min. after the treatment and one animal died 3 hours after the treatment.
All deaths seemed to be consequences of systemic toxic effect of the test item.
No death occurred at 50 mg/kg single oral dose of the test item. All female rats in step 3 and step 4 survived until the end of the 14-day observation period.
Clinical signs:
In group 1 treated with 2000 mg/kg bw dose:
Cclinical sign of reaction comprised of tonic convulsion (3 cases of 3 observations), clonic convulsion (3/3) and dyspnoea (3/3). These symptoms (score +4) occurred in all animals. These symptoms were detected on the treatment day up to 7 min. after the treatment.
In group 2 treated with 300 mg/kg bw dose:
Dyspnoea (score +2; +4) was observed in all animals. Tonic convulsion (score +4) and clonic convulsion (score +4) were detected in two animals. However, decreased activity (score -4), abnormal gait (score +3), blood around the nose (score +3), decreased grip- and limb tone (score -1), decreased body tone (score -1), decreased abdominal tone (score -1), decreased skin turgor (score -1), decreased ear reflex (score -1) and salivation (score +2; +3) was recorded in one animal. These symptoms were detected on the treatment day between 15 min. and 2 hours after the treatment.
In group 3 treated with 50 mg/kg bw dose:
Irritability (score +1; +2), abnormal gait (score +1; +2) and piloerection (score +1) were observed in all animals. However, decreased activity (score -1) was detected in one animal and increased activity (score +1) was recorded in one animal. These symptoms were detected on the treatment day and Day 1 post-dose.
In group 4 treated with 50 mg/kg bw dose:
Decreased activity (score -1), abnormal gait (score +1) and piloerection (score +1) were observed in all animals. Irritability (score +1) was recorded in two animals. However, restlessness was detected in one animal. These symptoms were detected on the treatment day.
Body weight:
The body weight and body weight gain data of group 1 (2000 mg/kg bw) and group 2 (300 mg/kg bw) could not be evaluated, because of mortalities.
In group 3 and 4 (50 mg/kg bw) the mean body weight of the animals corresponded to their species and age throughout the study.
Gross pathology:
All rats treated with 2000 mg/kg bw dose of the test item spontaneously died during the study and all rats treated with 300 mg/kg bw dose spontaneously died, too. No pathological changes were found related to the effect of the test item during the macroscopic examination of animals.
All animals treated with 50 mg/kg bw dose survived until the scheduled necropsy on Day 15. Slight hydrometra was recorded in one animal of the group 3 and moderate hydrometra was observed in one female of the group 4 is physiological finding and connected to the cycle of the animal.
No pathological changes were found related to the effect of the test item during the macroscopic examination of animals.

Any other information on results incl. tables

Groups

Treatment

Lethality

Test item

Dose (mg/kgbw)

Females

1

“2,3-dichlorobenzoyl nitrile
Step1

2000

3/3

2

“2,3-dichlorobenzoyl nitrile
Step2

300

3/3

3

“2,3-dichlorobenzoyl nitrile
Step3

50

0/3

4

“2,3-dichlorobenzoyl nitrile
Step4

50

0/3

 

Applicant's summary and conclusion

Interpretation of results:
Category 3 based on GHS criteria
Remarks:
Migrated information
Conclusions:
The method used is not intended to allow the calculation of a precise LD50 value.
The test item was ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423.
Hazard Category: Acute Tox. 3
Executive summary:

 Dose (mg/kg bw)

Mortality (dead/treated) 

LD50 (mg/kg bw) 

GHS category 

50

0/6 

between 50 and 300