Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 03 September 2013 (Start of in-life phase) to 11 December 2013 (GLP compliance statement)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
not specified
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
not specified
Qualifier:
according to
Guideline:
other: Food and Agricultural Materials Inspection Centre (FAMIC), 12 Nohsan, Notification No. 8147, April 2011; including the most recent partial revisions.
Deviations:
not specified
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: particulate/powder
Details on test material:
- Name of test material : Lithium cryolite
- Molecular formula : Li3AlF6
- Molecular weight : 162
- Physical state: White powder
- Storage condition of test material: At room temperature in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approx. 8 weeks old
-Health inspection : at least prior to dosing. It is ensured that the animals were healthy and without any abnormality that might affect the study integrity.
- Weight at study initiation: Mean of 151, 155 and 156 g for the group of 2000 mg/kg b.w. and the two additionnal group at 300 mg/kg bw respectively. Body weight variation did not exceed +/- 20% of the sex mean
- Fasting period : Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available ad libitum.
- Housing: Group housing of 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet : Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water : Free access to tap water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions.
Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): from 18 to 24 °C
- Humidity (%): from 40 to 70 %
- Air changes (per hr): approximately 15 room air changes/hour
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle

IN-LIFE DATES: From: 03 September 2013 To: 26 September 2013

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
VEHICLE
- Concentration in vehicle: The concentration of the test substance in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.
- Lot/batch no. (if required): No data
- Purity: Propylene glycol (Merck, Darmstadt, Germany) (specific gravity 1.036)

MAXIMUM DOSE VOLUME APPLIED:
10 mL/kg

CLASS METHOD
- Rationale for the selection of the starting dose: The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg. In a stepwise procedure two additional groups of three females were dosed at 300 mg/kg body weight. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.
Doses:
Single dosage on Day 1:
2000 mg/kg (10 mL/kg) body weight.
300 mg/kg (10 mL/kg) body weight.
No. of animals per sex per dose:
Each dose group consisted of 3 animals.:
2000 mg/kg bw : 3 animals
300 mg/kg bw : 3 + 3 animals
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability : Twice daily
Body weights: Days 1 (pre-administration), 8 and 15 and at death (if found dead or after Day 1)
Clinical signs : at periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded.
- Necropsy of survivors performed: The animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.
Statistics:
The oral LD50 value of the test substance was ranked within the following ranges: 0-5, 5-50, 50-300 or 300-2000 mg/kg b.w. or as exceeding 2000 mg/kg b.w. The LD50 cut-off value was established based on OECD guideline 423. No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Results and discussion

Preliminary study:
Not applicable
Effect levelsopen allclose all
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Key result
Sex:
female
Dose descriptor:
other: LD50 cut-off value
Effect level:
500 mg/kg bw
Based on:
test mat.
Remarks on result:
other: Defined according to the OECD test guideline 423.
Mortality:
At 2000 mg/kg all three animals dosed were found dead on Days 2, 4 and 6.
No further mortality occurred at 300 mg/kg.
Clinical signs:
At 2000 mg/kg hunched posture, piloerection, ptosis and/or hypothermia were noted among the majority of animals between Days 1 and 6.
At 300 mg/kg hunched posture and/or piloerection were noted among the animals on Days 1 and/or 2.
Body weight:
The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.
Gross pathology:
Macroscopic post mortem examination of one animal found dead revealed abnormalities of the stomach (isolated, dark red focus/foci in the glandular mucosa) and caecum (isolated, dark red focus/foci). Macroscopic post mortem examination of the other animals that died during the study and of the surviving animals at termination did not reveal any abnormalities.
Beginning of autolysis was seen in one animal found dead, this was considered not related to treatment.
Other findings:
No data.

Any other information on results incl. tables

Protocol deviations: there were no deviations from the protocol. There were no deviations from standard operating procedures that affected the integrity of the study.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The oral LD50 value in Wistar rats was established to be within the range of 300-2000 mg/kg body weight.
According to the OECD TG 423, the LD50 cut-off value was considered to be 500 mg/kg body weight.
The substance should be classified as harmful if swallowed according to the CLP and the UN GHS.
Executive summary:

The acute oral toxicity of Lithium cryolite in the rat was investigated according to the Acute Toxic Class Method of the OECD Testing Guideline 423 and under GLP.

Initially, Lithium cryolite was administered by oral gavage to three female Wistar rats at 2000 mg/kg body weight. In a stepwise procedure two additional groups of three females were dosed at 300 mg/kg body weight. All animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

At 2000 mg/kg all three animals dosed were found dead on Days 2, 4 and 6. No further mortality occurred at 300 mg/kg.

At 2000 mg/kg hunched posture, piloerection, ptosis and/or hypothermia were noted among the majority of animals between Days 1 and 6.

At 300 mg/kg hunched posture and/or piloerection were noted among the animals on Days 1 and/or 2.

The body weight gain shown by the animals over the study period was considered to be normal.

Macroscopic post mortem examination of one animal found dead revealed abnormalities of the stomach (isolated, dark red focus/foci in the glandular mucosa) and caecum (isolated, dark red focus/foci). Macroscopic post mortem examination of the other animals that died during the study and of the surviving animals at termination did not reveal any abnormalities.

The oral LD50 value of Lithium cryolite in Wistar rats was established to be within the range of 300-2000 mg/kg body weight.

According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight.

Based on these results, the substance should be classified as harmful if swallowed according to the CLP and the UN GHS.