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 and GLP compliant.

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-dichlorobenzoic acid (CAS: 50-45-3)
Batch No.: L2A134N
Physical state: Solid, crystalline powder
Colour: White
Active ingredient content (titrimetry): 100.4%
Water content: 0.03 % (m/m)
Storage: 15-30°C, protected from moisture

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
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): 162 - 167 g
Body weight range at starting (third step): 152 - 173 g

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: 8-12 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 two female animals died the test was continued at 300 mg/kg bw dose level on further three female rats. No animal died in the second step at 300 mg/kg bw dose level, three further female rats were treated with the same (300 mg/kg bw) dose. No animal died in the third 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 and 30 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 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 the surviver rats were sacrificed and necropsy were performed.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
300 - 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Two rats dosed at 2000 mg/kg bw “2,3-dichlorobenzoic acid” (CAS No. 50-45-3) died on Day 1. Both deaths seemed to be consequences of systemic toxic effect of the test item. The animal survived until the end of the 14-day observation period. No death occurred at 300 mg/kg single oral dose of the test item. All female rats in step 2 and step 3 survived until the end of the 14-day observation period.
Clinical signs:
In group 1 treated with 2000 mg/kg bw:
Decreased activity (score -1; -2; -3), pain reaction (score +1; +2), abnormal gait (score +1; +2), incoordination (score +1; +2; +3; +4), decreased grip- and limb tone (score -1; -2), decreased body tone (score -1; -2), decreased abdominal tone (score -1), decreased skin turgor (score -1) and piloerection (score +1; +2; +3) occurred in all animals. Decreased righting reflex (score -1) was observed in two animals. Blood around the nose (score +1) was detected in one animal. All animals were free of symptoms 30 min. after the treatment. The symptoms of toxicity started to exhibit themselves at the observation one hour after the treatment. The survivor animal become free of symptoms on Day 8 and remained symptom free till termination on Day 15.
In group 2 and 3 treated with 300 mg/kg bw:
No treatment related symptoms were observed throughout the 14-day post-treatment period.
Body weight:
In group 1 (2000 mg/kg bw) the body weight of the survivor animal corresponded to their species and age throughout the study.
In group 2 and 3 (300 mg/kg bw) the mean body weight of the animals corresponded to their species and age throughout the study.
Gross pathology:
Two rats treated with 2000 mg/kg bw dose of the test item spontaneously died during the study.
Slight hydrometra was observed in two females of the group 1 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.
All animals treated with 300 mg/kg bw dose survived until the scheduled necropsy on Day 15. Slight hydrometra was observed in one female of the group 2 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-dichlorobenzoicacid
Step1

2000

2/3

2

“2,3-dichlorobenzoicacid
Step2

300

0/3

3

“2,3-dichlorobenzoicacid
Step3

300

0/3

 

Applicant's summary and conclusion

Interpretation of results:
Category 4 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. 4
Executive summary:

 Dose (mg/kg bw) Mortality (dead/treated)  LD50 (mg/kg bw)  GHS category 
 300 0/6  between 300 and 2000