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Administrative data

Description of key information

In an acute oral toxicity study according to OECD guideline 423 the determined LD50 of the test item is 500 mg/kg bw in rats.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2020-03-20 to 2020-05-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001-12-17
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
2008-05-30
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002-12
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Remarks:
Han:WIST
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: Young adult rats, 10-11 weeks old
- Weight at study initiation: 196-214 g
- Fasting period before study: One day
- Housing: 3 animals/sex/cage, type III polypropylene/polycarbonate rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets
- Diet: Ad libitum (ssniff® SM R/M-Z+H complete diet, ssniff Spezialdiäten GmbH, D-59494 Soest Germany)
- Water: Ad libitum, tap water
- Acclimation period: 26 days in first group, 27 days in second group and 28 days in third group
- Microbiological status: SPF at arrival and kept in a good conventional environment during the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From day 0 to day 14
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
Aqua purificata
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 and 30 mg/mL
- Amount of vehicle: 10 mL/kg bw
- Batch no: 201910069
- Justification for choice of vehicle: Test item is soluble in vehicle.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD
- Rationale for the selection of the starting dose: The starting dose was selected on the basis of the available information about the test item.
Doses:
2000 mg/kg bw, 300 mg/kg bw
No. of animals per sex per dose:
3 (2000 mg/kg bw) and 6 (300 mg/kg bw)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Body weights: The body weight was recorded on day 0 (shortly before the treatment), on day 1, on day 7 and on day 15 on all animals with a precision of 1 g, respectively.
Signs of morbidity and mortality: Twice daily at the beginning and end of the working day.
Clinical Observations: Animals were observed individually after dosing once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h, after the treatment and once per day for 14 days thereafter.
- Necropsy of survivors performed: Yes
Statistics:
None
Key result
Sex:
female
Dose descriptor:
approximate LD50
Effect level:
ca. 500 mg/kg bw
Based on:
test mat.
Mortality:
Group 1 (2000 mg/kg bw): All female rats died on Day 1. All deaths might be a consequence of systemic toxic effects of the test item.
Group 2+3 (300 mg/kg bw): No mortality occurred. All rats in step 2 and step 3 survived until the end of the 14-day observation period.
Clinical signs:
Group 1 (2000 mg/kg bw): Decreased activity (15 cases out of 15 observations), incoordination (15/15), closed eyes (15/15) and piloerection (9/15). The score of symptoms was as follows: decreased activity (-3), incoordination (+3), closed eyes (+2) and piloerection (+2). These symptoms were detected on the treatment day between 30 minutes and 4 hours after the treatment. All symptoms were regarded as test item related.

Group 2 (300 mg/kg bw): Decreased activity (15 cases out of 57 observations), incoordination (15/57), closed eyes (15/57), piloerection (10/57) and salivation (1/57). Decreased activity (score -3, -2), incoordination (score +2, +1), closed eyes (score +2) and piloerection (score +1, +2) occurred in all animals. Salivation (score +2) was observed in one animal. These symptoms were detected on the treatment day between 30 minutes and 4 hours after the treatment. All symptoms were regarded as test item related.

Group 3 (300 mg/kg bw): Decreased activity (15 cases out of 57 observations), incoordination (15/57), closed eyes (15/57), piloerection (9/57) and salivation (2/57). Decreased activity (score -3, -2), incoordination (score +2, +1), closed eyes (score +2) and piloerection (score +3, +2) occurred in all animals. Salivation (score +1) was observed in two animals. These symptoms were detected on the treatment day between 30 minutes and 4 hours after the treatment. All symptoms were regarded as test item related.
Body weight:
Group 1 (2000 mg/kg bw): Not be evaluated, because all rats died on day 1.
Group 2+3 (300 mg/kg bw): The mean body weight and body weight gain of the animals corresponded to their species and age throughout the study.
Gross pathology:
Group 1 (2000 mg/kg bw): External macroscopic changes like bloody urethral orifice were found in all animals and blood around the nose in one animal. Internal macroscopic changes like stomach full of gas, attenuated stomach wall, yellowish-bloody content in the intestines and bloody urine in the urinary bladder were recorded in all animals. These internal macroscopic changes observed were regarded as test item related.

Group 2+3 (300 mg/kg bw): No pathological changes were found related to the effect of the test item during the macroscopic examination.

The method used is not intended to allow for the calculation of a precise LD50 value. However, the test item is ranked into the following category according to current EU Regulation on classification, labelling and packaging (CLP) (EC) No 1272/2008:





























Dose (mg/kg bw)



Mortality (dead/treated)



LD50 (mg/kg bw)



CLP category



2000



3/3 (at 1st step)



500



Category 4



300



0/6



Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
For this acute oral toxicity study with the test item, the determined LD50 is 500 mg/kg bw in rats.
Executive summary:

An acute oral toxicity study was carried out under GLP conditions using the class method according to OECD guideline 423. The method was conducted using a stepwise procedure with 2000 mg/kg bw as the starting dose in three female rats. The starting dose was selected on the basis of the available information about the test item. The test item was not expected to be lethal at 2000 mg/kg bw on the basis of safety data sheet (acute oral LD50 is 3050 mg/kg bw in rat). Since all animals died in 2000 mg/kg bw, a full test was performed. Since all animals died in the first step by day 1, further three female rats were treated with the lower (300 mg/kg bw) dose. No animal died in second step, and further three animals were treated with the same dose. No animal died in third step, too. The test was finished as a stopping criterion of Annex 2d of OECD Guideline No. 423 was met. Animals were weighed and observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out in animals that died on day 1, as well as in animals that survived until the end of the observation period on day 15.


 


Lethality, Clinical symptoms and Body weight:
All three animals treated with 2000 mg/kg bw of the test item died on day 1. No lethality was noted at a single oral dose of 300 mg/kg bw. In group 1, at a dose level of 2000 mg/kg bw, CNS-symptoms and abnormal movement (decreased activity, closed eyes), disturbances of the coordination (incoordination) and disturbance of the autonomic functions (piloerection) were observed in animals. These symptoms were detected on the treatment day between 30 minutes and 4 hours after the treatment. In group 2 and 3, at a dose level of 300 mg/kg bw, CNS-symptoms and abnormal movement (decreased activity, closed eyes), disturbance of the coordination (incoordination) and disturbances of the autonomic functions (piloerection, salivation) were observed in animals. These symptoms were detected on the treatment day between 30 minutes and 4 hours after the treatment. The body weight development was normal in all surviving animals.


 


Gross pathology:
Altogether 3 animals died, and 6 animals were sacrificed according to schedule at the end of the study period. External findings included bloody urethral orifice and blood around the nose. Internal findings included stomach full of gas, attenuated stomach wall, yellowish-bloody content in the intestines. Bloody urine in the urinary bladder was observed in group 1, as well. These findings were regarded as test item related. All of the animals treated with 300 mg/kg bw proved to be free of treatment related gross pathological changes.


 


Evaluation:


The method used is not intended to allow for the calculation of a precise LD50 value. However the test item is ranked into CLP category 4 according to current EU Regulation on classification, labelling and packaging (CLP) (EC) No 1272/2008. For this acute oral toxicity study with the test item in rats the determined LD50 is 500 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
500 mg/kg bw
Quality of whole database:
Reliable without restrictions

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Acute oral toxicity, key study


An acute oral toxicity study was carried out under GLP conditions using the class method according to OECD guideline 423. The method was conducted using a stepwise procedure with 2000 mg/kg bw as the starting dose in three female rats. The starting dose was selected on the basis of the available information about the test item. The test item was not expected to be lethal at 2000 mg/kg bw on the basis of safety data sheet (acute oral LD50 is 3050 mg/kg bw in rat). Since all animals died in 2000 mg/kg bw, a full test was performed. Since all animals died in the first step by day 1, further three female rats were treated with the lower (300 mg/kg bw) dose. No animal died in second step, and further three animals were treated with the same dose. No animal died in third step, too. The test was finished as a stopping criterion of Annex 2d of OECD Guideline No. 423 was met. Animals were weighed and observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out in animals that died on day 1, as well as in animals that survived until the end of the observation period on day 15.


 


Lethality, Clinical symptoms and Body weight:


All three animals treated with 2000 mg/kg bw of the test item died on day 1. No lethality was noted at a single oral dose of 300 mg/kg bw. In group 1, at a dose level of 2000 mg/kg bw, CNS-symptoms and abnormal movement (decreased activity, closed eyes), disturbances of the coordination (incoordination) and disturbance of the autonomic functions (piloerection) were observed in animals. These symptoms were detected on the treatment day between 30 minutes and 4 hours after the treatment. In group 2 and 3, at a dose level of 300 mg/kg bw, CNS-symptoms and abnormal movement (decreased activity, closed eyes), disturbance of the coordination (incoordination) and disturbances of the autonomic functions (piloerection, salivation) were observed in animals. These symptoms were detected on the treatment day between 30 minutes and 4 hours after the treatment. The body weight development was normal in all surviving animals.


 


Gross pathology:


Altogether 3 animals died, and 6 animals were sacrificed according to schedule at the end of the study period. External findings included bloody urethral orifice and blood around the nose. Internal findings included stomach full of gas, attenuated stomach wall, yellowish-bloody content in the intestines. Bloody urine in the urinary bladder was observed in group 1, as well. These findings were regarded as test item related. All of the animals treated with 300 mg/kg bw proved to be free of treatment related gross pathological changes.


 


Evaluation:


The method used is not intended to allow for the calculation of a precise LD50 value. However the test item is ranked into CLP category 4 according to current EU Regulation on classification, labelling and packaging (CLP) (EC) No 1272/2008. For this acute oral toxicity study with the test item in rats the determined LD50 is 500 mg/kg bw.

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No 1272/2008


The available test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Thus, the test item is considered to be classified for acute oral toxicity (category 4) under Regulation (EC) No 1272/2008, as amended for fifteenth time in Regulation (EU) No 2020/1182.