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Description of key information

Acute Oral Toxicity testing according to the Acute Toxic Class Method resulted to an LD50 between 200 and 2000 mg/kg bw in rats, with a LD50 cut-off level of 1000 mg/kg bw.

No acute dermal and inhalation toxicity studies were performed on the substance due to its corrosive properties.

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:
from 2003-09-17 to 2003-10-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study, although OECD 423 version of 1996 rather than the updated version of 2001.
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: 8 weeks old
- Weight at study initiation: 270 ± 10 g for the males, and 222 ± 3 g for the females
- Fasting period before study: overnight pre-testing fasting
- Housing: in polycarbonate cages with stainless steel lids. Each cage contained 1 to 7 rats of the same sex during acclimation, and 3 rats of the same sex and group during the treatment period.
- Diet : ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±2°C
- Humidity (%): 30 to 70%
- Air changes (per hr): 12 hour light/dark cycle
- Photoperiod (hrs dark / hrs light): 12 air changes per hour.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentrations in vehicle: 20 and 200 mg/ml
- Amount of vehicle (if gavage): 10 ml/kg
- Justification for choice of vehicle: solubility

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

CLASS METHOD
- Rationale for the selection of the starting dose:
the dose-level used as the starting dose was selected from one of the 3 fixed levels: 25, 200 or 2000 mg/kg bw. As no information on the toxic potential of the test item was available, for animal welfare reasons, the starting dose of 200 mg/kg bw was chosen
Doses:
200 and 2000 mg/kg/bw
No. of animals per sex per dose:
3 males and females given 200 mg/kg bw, 3 males given 2000 mg/kg bw
Control animals:
other: Historical data of control animals dosed by oral route
Details on study design:
- Duration of observation period following administration: 14 days after first day of dosing (D1)

- Frequency of observations and weighing: the animals were observed frequently during the hours following administration of the test item, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day. Type, time of onset and duration of clinical signs were recorded for each animal individually. The animals were weighed individually just before administration of the test item on day 1 and then on days 8 and 15.

- Necropsy of survivors performed: yes
Sex:
male
Dose descriptor:
LD50
Effect level:
200 - 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: toxicity is comparable in females
Mortality:
No mortality occurred at the 200 mg/kg dose in both sexes. At the 2000 mg/kg dose-level, 2out of 3 males were found dead on day 2 or 3.
Clinical signs:
At 200 mg/kg:
Hypoactivity, piloerection and dyspnea, together with hypersalivation on day 1, were observed in all males on days 1 and 2. In all females, only hypoactivity and piloerection were noted on day 1.
At 2000 mg/kg:
Hypoactivity, piloerection and dyspnea were recorded prior to death. In the surviving male, the same clinical signs, together with swollen abdomen and loud breathing on day 3, were observed from day 1 up to day 3; loud breathing persisted until day 12.
Body weight:
The overall body weight gain of the animals given 200 mg/kg was similar to that of CIT historical control animals.
The body weight gain of the surviving male given 2000 mg/kg was reduced during the first week of the study when compared to CIT historical control animals.
Gross pathology:
Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.
Interpretation of results:
other: Toxicity category 4 / Harmful
Remarks:
Criteria used for interpretation of results: other: CLP (Reg. 1272/2008/EC) + expert judgement and Directive 67/548/EEC
Conclusions:
The oral LD50 of the test item was comprised between 200 and 2000 mg/kg in rats, with a LD50 cut-off of 1000 mg/kg.
Executive summary:

The acute oral toxicity of the test item was evaluated in rats according to OECD guideline 423 (1996) and EU method B.1tris. The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.  

The test item was prepared in corn oil and was administered by oral route (gavage), under a volume of 10 mL/kg, to groups of three fasted Sprague-Dawley rats. Males were used in the initial step starting with a dose-level of 200 mg/kg bw. As no deaths occured, another assay was

carried out on 3 other males at 2000 mg/kg bw . After this second assay as 2/3 animals died, another assay was carried out on 3 females at the next lower dose-level of 200 mg/kg bw.

At each step, clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test item. All animals were subjected to necropsy.

At the 200 mg/kg bw dose-level, no mortality occured. Hypoactivity, piloerection and dyspnea, together with hypersalivation on day 1, were observed in all males on days 1 and 2. In all females, only hypoactivity and piloerection were noted on day 1. When compared to historical control data, the body weight gain of the animals was not affected by treatment with the test item. At necropsy, no apparent abnormalities were observed in any animal.

At the 2000 mg/kg bw dose-level, 2out of 3 males were found dead on day 2 or 3. Hypoactivity, piloerection and dyspnea were recorded prior to death. In the surviving male, the same clinical signs, together with swollen abdomen and loud breathing on day 3, were observed from day 1 up to day 3; loud breathing persisted until day 12. The body weight gain of the surviving male given 2000 mg/kg was reduced during the first week of the study when compared to historical control animals. At necropsy, no apparent abnormalities were observed.

Under these experimental conditions, the oral LD50of the test item was comprised between 200 and 2000 mg/kg in male rats. Toxicity was comparable in females. The LD50 cut-off is established at 1000 mg/kg.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 000 mg/kg bw
Quality of whole database:
High quality study, showing highest toxicity of availabe studies.

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:

One reliable without restriction study was reported for the acute oral toxicity endpoint and was selected as a key study fro classification.

 

The study of Ollivier (2004), was designed to evaluate the acute oral toxicity of the test item in rats according to OECD guideline 423 (1996) and EU method B.1tris (1996). The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.  

The test item was prepared in corn oil and was administered by oral route (gavage), under a volume of 10 mL/kg, to groups of three fasted Sprague-Dawley rats. Males were used in the initial step starting with a dose-level of 200 mg/kg bw. As no deaths occurred, another assay was

carried out on 3 other males at 2000 mg/kg bw . After this second assay as 2/3 animals died, another assay was carried out on 3 females at the next lower dose-level of 200 mg/kg bw.

At each step, clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test item. All animals were subjected to necropsy.

At the 200 mg/kg bw dose-level, no mortality occurred. Hypoactivity, piloerection and dyspnea, together with hypersalivation on day 1, were observed in all males on days 1 and 2. In all females, only hypoactivity and piloerection were noted on day 1. When compared to historical control data, the body weight gain of the animals was not affected by treatment with the test item. At necropsy, no apparent abnormalities were observed in any animal.

At the 2000 mg/kg bw dose-level, 2 out of 3 males were found dead on day 2 or 3. Hypoactivity, piloerection and dyspnea were recorded prior to death. In the surviving male, the same clinical signs, together with swollen abdomen and loud breathing on day 3, were observed from day 1 up to day 3; loud breathing persisted until day 12.The body weight gain of the surviving male given 2000 mg/kg was reduced during the first week of the study when compared to historical control animals. At necropsy, no apparent abnormalities were observed.

Under these experimental conditions, the oral LD50of the test item was comprised between 200 and 2000 mg/kg in male rats. Toxicity was comparable in females. This results to a LD50 cut-off level is 1000 mg/kg bw.

 

Another acute oral toxicity study conducted in rats in the eighties was not taken into account for the classification of the substance. The document showed some insufficiencies for assessment.

In the dose range finding for this study mortality was 0/2, 0/2, 0/2, 0/2 and 1/2 in the 250, 500, 1000, 3000 and 5000 mg/kg bw dose groups respectively. In the main study groups of 5 males and 5 females were dosed levels of 3000, 4000 and 5000 mg/kg in dose volumes of 20 mL/kg bw of distilled water. Observed mortality rates were 3/5, 3/5 and 3/5 in males and 0/5, 3/5 and 4/5 in females.

LD50 was calculated based on assumption of no mortality in males at 1000 mg/kg due to the constant mortality of 3 out of 5 males observed at all tested dose levels (Cuthbert, 1983).

From the mortality pattern in the main study and the assumption of no mortaility for the males at 1000 mg/kg the LD50's with 95% confidence limits were calculated to be: 3610 (2919 - 4464) mg/kg bw in male rats and 4260 (3975 - 4565) mg/kg bw in female rats. When taking all male and female mortality rates together, including the range finding, LD50 would be between 3000 (mortality 25%) and 4000 mg/kg (mortality 60%).

The higher levels of the LD50 observed can possibly be partly explained by the lower concentrations (20 mL vs 10 mL/kg bw) and the use of water versus oil. When comparing the actual mortalities of 2/3 males at 2000 mg/kg (Olivier), with the 3/5 males at 3000 mg/kg (Cuthbert) and considering the low slope seen in the Cuthbert study, the results of these studies are not that much different in results.

 

 

Acute dermal and inhalation toxicity:

The substance is classified as corrosive to skin and testing for acute toxicity is therefore not needed according to REACH regulation (EC) 1907/2006 (Annex VII, point 8.5, column 2).

Justification for selection of acute toxicity – oral endpoint

Most reliable study available.

Justification for selection of acute toxicity – inhalation endpoint

The substance is classified as corrosive and no acute toxicity testing should normally be conducted. As the substance is corrosive, symptoms of local respiratory irritation are expected, which should limit the systemic uptake of amount needed for systemic toxicity considering the relatively low acute oral toxicity

Justification for selection of acute toxicity – dermal endpoint

In accordance with column 2 of REACH Annex VIII, the acute dermal toxicity study does not need to be conducted as the substance is classified as corrosive to the skin.

Justification for classification or non-classification

Oral route: The results of the study conducted by Ollivier (2004) resulted to a LD500 cut-off level of 1000 mg/kg. According to the criteria laid down in EU directive67/548/EEC, the substance is therefore classified Harmful with the risk phrase R22.

According to the criteria laid down in EU regulation (EC) n° 1272/2008/EC (CLP), the substance should be classified in category 4 of toxicity with the hazard statement H302.

 

Dermal route: Acute oral toxicity testing indicates a relatively low acute systemic toxicity, and effects observed are attributable to its corrosive properties in the GI-tract. Following its severe corrosive properties, systemic toxicity following acute dermal exposure is not likely to occur. Therefore no classification is warranted.

 

Inhalation route: No data is available on inhalation toxicity. The combination of very low vp (< 3 x 10-7 Pa), low likelihood of exposures aerosols and the severe corrosive local effects expected for this substance makes further acute toxicity testing by inhalation with the substance itself ethically not justified.

Based on its physical appearance as viscous liquid with solid parts (viscosity 500 mPa.s at 20°C), there is no need for classification for aspiration hazard.

 

The available data does not show indication that classification for STOT-SE cat 1 or 2 is indicated.

For STOT-SE Cat 3: There is no indication for possible narcotic effects. Although the substance is corrosive, the very low vapour pressure prohibits occurrence of respiratory irritation by vapour. The classification of corrosive is already considered to implicitly cover the potential of RTI and additional Cat.3 is considered to be superfluous (Guidance CLP Ch. 3.8.2.5)