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Description of key information

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 respectively. 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.

 

The acute inhalation toxicity of Lithium cryolite in the rat was investigated according to the OECD Testing Guideline 403 and under GLP.

Lithium cryolite was administered as an aerosol by inhalation for 4 hours to one group of five male and five female Wistar rats at 1 mg/L. Based on the results, one group of five females was dosed at 0.5 mg/L. Animals were subjected to daily observations and determination of body weights on Days 1, 2, 4, 8 and 15 and at death. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

For the exposure to 1 mg/L, the time-weighted mean actual concentration was 1.1 ± 0.05 mg/L. For the exposure to 0.5 mg/L, the time-weighted mean actual concentration was 0.52 ± 0.01 mg/L. The concentration measurements distributed over time showed that the substance was sufficiently stable.

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice. At 1 mg/L, the MMAD was 2.9 µm (gsd 2.0) and 2.6 µm (gsd 2.0). At 0.5 mg/L, the MMAD was 3.6 µm (gsd 1.9) and 3.2 µm (gsd 1.8).

At 1 mg/L, two females were found dead on Days 3 and 4. Two males and three females were sacrificed for ethical reasons on days 4 and 5. At 0.5 mg/L, one female was sacrificed on day 5. No further mortality occurred.

At 1 mg/L, slow breathing was noted among the animals during exposure. After exposure, lethargy, flat posture, hunched posture, slow breathing, laboured respiration, shallow respiration, piloerection, chromodacryorrhoea, ptosis and or hypothermia were noted among the animals. The surviving animals recovered from the symptoms between Days 7 and 9. At 0.5 mg/L, no clinical signs were seen during exposure. After exposure, abnormal posture, hunched posture, piloerection, dehydration, lean appearance and/or ptosis were seen among the animals. The surviving animals had recovered from the symptoms by Day 11.

Body weight loss was noted among all surviving animals during the first week post-exposure. All animals regained weight during the second week.

Macroscopic post mortem examination revealed abnormalities of the thymes (reduced in size) of one female (0.5 mg/L) sacrificed for ethical reasons during the study. No other findings were noted in any of the animals.

Based on the above observations, the inhalatory LC50, 4h value of LITHIUM CRYOLITE in Wistar rats was established to be within the range of 0.5 – 1 mg/L.

 

The acute dermal toxicity of Lithium cryolite in the rat was investigated according to the OECD Testing Guideline 402 and under GLP.

Lithium cryolite was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred.

Chromodacryorrhoea was noted for all animals on Days 1 and 2. General erythema, erythema maculate, scales and/or scabs were noted among the female animals between Days 3 and 15.

The mean body weight gain during the observation period was within the range expected for rats used in this type of study.

No abnormalities were found at macroscopic post mortem examination of the animals.

The dermal LD50 value of Lithium cryolite in Wistar rats was established to exceed 2000 mg/kg body weight.

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 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
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
not specified
Qualifier:
according to guideline
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. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system 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
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).
Preliminary study:
Not applicable
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.

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

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.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
500 mg/kg bw
Quality of whole database:
One reliable key study is available for the acute oral toxicity of Lithium cryolite in the rat, performed according to the Acute Toxic Class Method of the OECD Testing Guideline 423 and under GLP.
The oral LD50 value in Wistar rats was established to be within the range of 300-2000 mg/kg body weight.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 23 September 2013 (Start of in-life phase) to 20 December 2013 (GLP compliance statement)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Deviations:
not specified
Qualifier:
according to guideline
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. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
Health inspection: At least prior to exposure. It was ensured that the animals were healthy and without any abnormality that might affect the study integrity.
- Age at study initiation: Young adult animals were selected (approximately 10-11 weeks old).
- Weight at study initiation: Animals used within the study were of approximately the same age and body weight variation did not exceed +/- 20% of the sex mean.
- Fasting period before study: none
- Housing: Group housing of five animals per sex per cage in labelled Makrolon cages (type IV; height 18 cm) containing sterilised 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).
Animal husbandry on the Day of exposure : The animals were moved to the inhalation area to in order to perform the exposure. During the exposure, there was no access to food and water. After exposure, the animals were returned their cages which were placed in a fume cupboard for a short time period to allow test substance remnants to evaporate. A sheet of filter paper was used to cover the bedding material to prevent suffocation in case of bad health condition and in order to recover and to aid the clinical observations. The sheet was removed and before the end of the exposure day, the animals were returned the animal room.
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) except during exposure to the test substance.
- Water (e.g. ad libitum): Free access to tap water except during exposure to the test substance.
- Acclimation period: Acclimatisation period was 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): Environmental controls for the animal room were set to maintain 18 to 24°C
- Humidity (%): a relative humidity of 40 to 70%
- Air changes (per hr): approximately 15 room air changes/hour
- Photoperiod (hrs dark / hrs light): a 12-hour light/12-hour dark cycle
Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.


IN-LIFE DATES: From: 23 September 2013 To: 17 October 2013
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure chamber:
The design of the exposure chamber is based on the flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983). The chamber consisted of three animal sections with eight animal ports each. Each animal port had its own atmosphere inlet and exhaust outlet. The animals were placed in restraining tubes and connected to the animal ports. The number of animal sections and number of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 1 L/min, which ensures an adequate oxygen supply to the animals. The main inlet of the test atmosphere was located at the top section and the main outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal.
All components of the exposure chamber in contact with the test material were made of stainless steel, glass, rubber or plastic. To avoid exposure of the personnel and contamination of the laboratory the exposure chamber was placed in a fume hood, which maintained at a slight negative pressure.

- Test atmosphere generation :
Administering the test substance to a stream of pressurized air using a combination of a spiral feeder and micronizing jet mill (Bernstein, D.N., Aerosols, pp 721- 723, 1984) generated an aerosol. The rotation speed of the feeder was varied to obtain the desired exposure concentration. The aerosol was passed through a series of cyclones, allowing larger particles to settle. The primary aerosol was diluted with pressurized air before it entered the exposure chamber. The mean total airflows were 91 and 98 L/min. for the 1 and 0.5 mg/L exposure groups respectively.
From the exposure chamber the test atmosphere was passed through a filter before it was released to the exhaust of the fume hood.

- Method of holding animals in test chamber:
see section "details on test animals and environmental conditions".

- Method of particle size determination:
The particle size distribution was characterized twice during each exposure period. The samples were drawn (2 L/min) from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber. The samples were collected with an 8 stage Marple personal cascade impactor containing fiber glass filters (SKC 225-713, fiber glass, SKC Omega Specialty Division, Chelmsford, MA, USA) and a fiber glass back-up filter (SEC-290-F1, Westech, Upper Stondon, Bedfordshire, England). Amounts of test substance collected were measured gravimetrically. Subsequently the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD) were determined.

- Temperature, humidity, pressure in air chamber:
The temperature and relative humidity were measured with a humidity and temperature indicator (E+E Elektronik, Engerwitzdorf, Austria) and were recorded after the animals were placed in the experimental set-up and at 30 minute intervals after initiation of the exposure. The probe was inserted in a tube mounted in one of the free animal ports of the middle section of the exposure chamber.
The temperature of the atmosphere during the exposures was between 20.7 and 21.8°C and relative humidity was between 27 and 39%. These conditions were considered appropriate for this relatively short 4 hours exposure duration.


TEST ATMOSPHERE
- Brief description of analytical method used:
A total of 16 and 21 representative samples were taken for determination of the actual concentration during exposure at 1 and 0.5 mg/L, respectively. Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber.
Samples were drawn through a glass fiber filter (type SF 92 vorfilter, Schleicher & Schuell, Dassel, Germany). The collected amount of the test substance in the air sample was measured gravimetrically. Sample volumes were measured by means of a dry gas meter (type G 1.6, Actaris Meterfabriek B.V., Dordrecht, The Netherlands).
Subsequently the time-weighted mean concentrations with the standard deviations were calculated.
It was considered during the trial generations that the opacity of the test atmosphere could not be reliably monitored by means of an aerosol monitoring system. An indication of the stability of the test atmosphere was obtained from the concentration measurements, which were equally distributed over time.

- Samples taken from breathing zone: yes

VEHICLE
No vehicle used except air.

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: see section "Any other information on results incl. tables"
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice. At 1 mg/L, the MMAD was 2.9 µm (gsd 2.0) and 2.6 µm (gsd 2.0). At 0.5 mg/L, the MMAD was 3.6 µm (gsd 1.9) and 3.2 µm (gsd 1.8).
CLASS METHOD
- Rationale for the selection of the starting concentration: Target concentrations were based on the cut off concentration values specified in the UN and EC classification guidelines. Five animals of each sex were exposed in a limit test for 4 hours to a target concentration of the test substance of 1 mg/L. Based on the results, five females (identified as most sensitive sex) were exposed to 0.5 mg/L.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
gravimetrically measurement
Duration of exposure:
4 h
Concentrations:
0.5 and 1 mg/L.

The nominal concentration was calculated by dividing the amount of test substance used by the volume of pressurized air (average air flow times exposure time) entering the exposure chamber used for exposure of the animals. Due to the small volume of the exposure chamber the equilibrium time was negligible. The volume of air was calculated from the average air flow (measured by means of thermal mass flow meters and was recorded regularly, preferably in 30 minute intervals) and the exposure time.
No. of animals per sex per dose:
5 animals of each sex were exposed to a target concentration of the test substance of 1 mg/L.
5 females (identified as most sensitive sex) were exposed to 0.5 mg/L.
Control animals:
no
Details on study design:
- Treatment:
Prior to each exposure, both eyes of each rat were instilled with Opthosan (AST Farma BV, Oudewater, The Netherlands) to protect the eyes against potential irritation by the test substance. Prior to exposure the animals were restrained in polycarbonate restraining tubes; these tubes were connected to the exposure chamber. Sixteen minutes after the last animal was placed the generation of the test atmosphere was started. The exposure time was 4 hours.

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
Mortality/Viability: Twice daily. The time of death was recorded as precisely as possible.
Body weights: Days 1 (pre-administration), 2, 4, 8 and 15 and at death (if found dead or sacrificed after Day 1).

- Necropsy of survivors performed: yes
The moribund animals and animals surviving to the end of the observation period were sacrificed by an intraperitoneal injection with Euthasol ® (AST Farma BV, Oudewater, The Netherlands). All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded. Particular attention was given to any changes in the respiratory tract.

- Other examinations performed:
clinical signs during exposure: three times during exposure for mortality, behavioural signs of distress and effects on respiration.
clinical signs after exposure : On Day 1, one and three hours after exposure 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:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).

- Electronic capture data :
Observations/measurements in the study were recorded electronically using the following programs:
REES Centron Environmental Monitoring system version SQL 2.0 (REES Scientific, Trenton, NJ, USA): Environmental monitoring.
TOXDATA version 8.0 (WIL Research Europe B.V., ‘s-Hertogenbosch, The Netherlands): Mortality / Clinical signs / Body weights. Clinical signs during exposure or not defined in TOXDATA and body weights of decedent animals were recorded manually.
Statistics:
No statistical analysis was performed.
Preliminary study:
No preliminary study has been performed.
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 0.5 - < 1 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: only females were exposed at 0.5 mg/L because identified as the most sensitive sex at 1 mg/L
Mortality:
At 1 mg/L, two females were found dead on Days 3 and 4. Two males and three females were sacrificed for ethical reasons on days 4 and 5.
At 0.5 mg/L, one female was sacrificed on day 5. No further mortality occurred.
Clinical signs:
other: At 1 mg/L, slow breathing was noted among the animals during exposure. After exposure, lethargy, flat posture, hunched posture, slow breathing, laboured respiration, shallow respiration, piloerection, chromodacryorrhoea, ptosis and or hypothermia were not
Body weight:
Body weight loss was noted among all surviving animals during the first week post-exposure. All animals regained weight during the second week.
Gross pathology:
Macroscopic post mortem examination revealed abnormalities of the thymes (reduced in size) of one female (0.5 mg/L) sacrificed for ethical reasons during the study. No other findings were noted in any of the animals.

List of deviations

- protocol deviations : deviations from the maximum level for daily mean relative humidity occurred.

Evaluation: Laboratory historical data do not indicate an effect of the deviations. The study integrity was not adversely affected by the deviation.

- standard operating procedures deviations : none.

Test atmosphere characterization : Concentration

For the exposure to 1 mg/L, the time-weighted mean actual concentration was 1.1 ± 0.05 mg/L. The nominal concentration (amount of test substance used divided by the volume of pressurized air used) was 8.2 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 13%.

For the exposure to 0.5 mg/L, the time-weighted mean actual concentration was 0.52 ± 0.01 mg/L. The nominal concentration (amount of test substance used divided by the volume of pressurized air used) was 0.47 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 111%.

The concentration measurements distributed over time showed that the substance was sufficiently stable.

1 mg/L exposure group

Start of generation of test atmosphere:

08:50

 

End of generation of test atmosphere:

12:50

 

Time

(hh:mm)

Action

Sample volume

(L)

Mass sampled (mg)

Concentration

(mg/L)

 

 

 

% of total

exposure

time

 

 

 

Weight

concentration

(mg/L)

8:50

start of generation

n.a.

n.a.

n.a.

 

0.0

 

n.a.

8:52

start of sampling

5

6.37

1.274

 

0.8

 

0.011

9:02

start of sampling

5

6.19

1.238

 

4.2

 

0.052

9:12

start of sampling

5

6.73

1.346

 

4.2

 

0.056

9:19

start of sampling

5

5.00

1.000

 

2.9

 

0.029

9:34

start of sampling

5

4.30

0.860

 

6.2

 

0.054

9:44

start of sampling

5

6.43

1.286

 

4.2

 

0.054

10:12

start of sampling

5

5.68

1.136

 

11.7

 

0.133

10:32

start of sampling

5

6.03

1.206

 

8.3

 

0.101

11:07

start of sampling

5

6.50

1.300

 

14.6

 

0.190

11:16

start of sampling

5

5.92

1.184

 

3.8

 

0.044

11:26

start of sampling

5

3.32

0.664

 

4.2

 

0.028

11:31

start of sampling

5

5.30

1.060

 

2.1

 

0.022

11:51

start of sampling

5

4.84

0.968

 

8.3

 

0.081

12:13

start of sampling

5

6.32

1.264

 

9.2

 

0.116

12:29

start of sampling

5

5.30

1.060

 

6.7

 

0.071

12:40

start of sampling

5

4.98

0.996

 

4.6

 

0.046

12:50

end of generation

n.a.

n.a.

0.996

 

4.2

 

0.041

 

 

 

Time-weighted mean concentration

1.126

 

 

 

 

 

Standard deviation

0.046

 

 

 

 

 

n

16

 

 

0.5 mg/L exposure group

Start of generation of test atmosphere:

08:41

 

End of generation of test atmosphere:

12:41

 

Time

(hh:mm)

Action

Sample volume

(L)

Mass sampled (mg)

Concentration

(mg/L)

 

 

 

% of total

exposure

time

 

 

 

Weight

concentration

(mg/L)

8:41

start of generation

n.a.

n.a.

n.a.

 

0.0

 

n.a.

8:44

start of sampling

5

3.07

0.614

 

1.3

 

0.008

8:50

start of sampling

5

4.46

0.892

 

2.5

 

0.022

8:55

start of sampling

5

2.43

0.486

 

2.1

 

0.010

9:05

start of sampling

5

1.81

0.362

 

4.2

 

0.015

9:11

start of sampling

5

2.22

0.444

 

2.5

 

0.011

9:18

start of sampling

5

3.10

0.620

 

2.9

 

0.018

9:24

start of sampling

5

2.34

0.468

 

2.5

 

0.012

9:35

start of sampling

5

2.27

0.454

 

4.6

 

0.021

9:47

start of sampling

5

2.03

0.406

 

5.0

 

0.020

9:52

start of sampling

5

2.08

0.416

 

2.1

 

0.009

9:58

start of sampling

5

2.88

0.576

 

2.5

 

0.014

10:17

start of sampling

5

2.56

0.512

 

7.9

 

0.041

10:36

start of sampling

6

3.47

0.578

 

7.9

 

0.046

10:44

start of sampling

5

2.54

0.508

 

3.3

 

0.017

10:58

start of sampling

5

2.56

0.512

 

5.8

 

0.030

11:20

start of sampling

5

2.62

0.524

 

9.2

 

0.048

11:39

start of sampling

5

2.72

0.544

 

7.9

 

0.043

11:46

start of sampling

5

2.41

0.482

 

2.9

 

0.014

12:04

start of sampling

5

2.63

0.526

 

7.5

 

0.039

12:17

start of sampling

5

2.39

0.478

 

5.4

 

0.026

12:29

start of sampling

5

2.81

0.562

 

5.0

 

0.028

12:41

end of generation

n.a.

n.a.

0.562

 

5.0

 

0.028

 

 

Time-weighted mean concentration

0.520

 

 

 

 

 

Standard deviation

0.013

 

 

 

 

 

n

21

 

1)    Assumed concentration, based on the last sample.

2)    n.a.= not applicable

Test atmosphere characterization: Particle size

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice. At 1 mg/L, the MMAD was 2.9 µm (gsd 2.0) and 2.6 µm (gsd 2.0). At 1 mg/L, the MMAD was 3.6 µm (gsd 1.9) and 3.2 µm (gsd 1.8).

1 mg/L exposure group

 

 

 

Start of generation of test atmosphere:

08:34

End of generation of test atmosphere:

12:50

Sampling speed (L/min):

2

measurement 1:

Sampling time:

09:52

Sample volume (L):

7

 

Stage

Cut point

(mm)

Mass sampled

(mg)

Relative mass

(%)

Cumulative mass

(% of total sampled)

1

21.0

0.03

0.47

99.53

2

15.0

0.07

1.09

98.45

3

10.0

0.08

1.24

97.20

4

6.0

0.65

10.09

87.11

5

3.5

1.33

20.65

66.46

6

2.0

2.81

43.63

22.83

7

0.9

1.03

15.99

6.83

8

0.5

0.44

6.83

0.00

Back up

0.25

0.00

0.00

0.00

MMAD1(mm):

2.9

gsd2:

2.0

1 Mass Median Aerodynamic Diameter

2 Geometric standard deviation

 

measurement 2:

Sampling time:

11:42

Sample volume (L):

7

 

Stage

Cut point

(mm)

Mass sampled

(mg)

Relative mass

(%)

Cumulative mass

(% of total sampled)

1

21.0

0.03

0.49

99.51

2

15.0

0.03

0.49

99.02

3

10.0

0.02

0.33

98.69

4

6.0

0.48

7.83

90.86

5

3.5

2.15

35.07

55.79

6

2.0

2.23

36.38

19.41

7

0.9

0.74

12.07

7.34

8

0.5

0.43

7.01

0.33

Back up

0.25

0.02

0.33

0.00

MMAD1(mm):

2.6

gsd2:

2.0

1 Mass Median Aerodynamic Diameter

2 Geometric standard deviation

 

0.5 mg/L exposure group

 

 

Start of generation of test atmosphere:

08:41

End of generation of test atmosphere:

12:41

Sampling speed (L/min):

2

measurement 1:

Sampling time:

10:02

Sample volume (L):

16

 

Stage

Cut point

(mm)

Mass sampled

(mg)

Relative mass

(%)

Cumulative mass

(% of total sampled)

1

21.0

0.08

1.94

98.06

2

15.0

0.08

1.94

96.12

3

10.0

0.14

3.40

92.72

4

6.0

0.31

7.52

85.19

5

3.5

2.02

49.03

36.17

6

2.0

0.82

19.90

16.26

7

0.9

0.54

13.11

3.16

8

0.5

0.13

3.16

0.00

Back up

0.25

0.00

0.00

0.00

MMAD1(mm):

3.6

gsd2:

1.9

1 Mass Median Aerodynamic Diameter

2 Geometric standard deviation

 

measurement 2:

Sampling time:

11:53

Sample volume (L):

16

 

Stage

Cut point

(mm)

Mass sampled

(mg)

Relative mass

(%)

Cumulative mass

(% of total sampled)

1

21.0

0.00

0.00

100.00

2

15.0

0.02

0.53

99.47

3

10.0

0.02

0.53

98.94

4

6.0

0.30

7.98

90.96

5

3.5

2.39

63.56

27.39

6

2.0

0.61

16.22

11.17

7

0.9

0.29

7.71

3.46

8

0.5

0.13

3.46

0.00

Back up

0.25

0.00

0.00

0.00

MMAD1(mm):

3.2

gsd2:

1.8

1 Mass Median Aerodynamic Diameter

2 Geometric standard deviation

Interpretation of results:
Category 3 based on GHS criteria
Conclusions:
The inhalatory LC50, 4h value of LITHIUM CRYOLITE in Wistar rats was established to be within the range of 0.5 – 1 mg/L.
Executive summary:

The acute inhalation toxicity of Lithium cryolite in the rat was investigated according to the OECD Testing Guideline 403 and under GLP.

 

Lithium cryolite was administered as an aerosol by inhalation for 4 hours to one group of five male and five female Wistar rats at 1 mg/L. Based on the results, one group of five females was dosed at 0.5 mg/L. Animals were subjected to daily observations and determination of body weights on Days 1, 2, 4, 8 and 15 and at death. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

 

For the exposure to 1 mg/L, the time-weighted mean actual concentration was 1.1 ± 0.05 mg/L. For the exposure to 0.5 mg/L, the time-weighted mean actual concentration was 0.52 ± 0.01 mg/L. The concentration measurements distributed over time showed that the substance was sufficiently stable.

 

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice. At 1 mg/L, the MMAD was 2.9 µm (gsd 2.0) and 2.6 µm (gsd 2.0). At 0.5 mg/L, the MMAD was 3.6 µm (gsd 1.9) and 3.2 µm (gsd 1.8).

 

At 1 mg/L, two females were found dead on Days 3 and 4. Two males and three females were sacrificed for ethical reasons on days 4 and 5. At 0.5 mg/L, one female was sacrificed on day 5. No further mortality occurred.

 

At 1 mg/L, slow breathing was noted among the animals during exposure. After exposure, lethargy, flat posture, hunched posture, slow breathing, laboured respiration, shallow respiration, piloerection, chromodacryorrhoea, ptosis and or hypothermia were noted among the animals. The surviving animals recovered from the symptoms between Days 7 and 9. At 0.5 mg/L, no clinical signs were seen during exposure. After exposure, abnormal posture, hunched posture, piloerection, dehydration, lean appearance and/or ptosis were seen among the animals. The surviving animals had recovered from the symptoms by Day 11.

 

Body weight loss was noted among all surviving animals during the first week post-exposure. All animals regained weight during the second week.

 

Macroscopic post mortem examination of revealed abnormalities of the thymes (reduced in size) of one female (0.5 mg/L) sacrificed for ethical reasons during the study. No other findings were noted in any of the animals.

 

Based on the above observations, the inhalatory LC50, 4h value of LITHIUM CRYOLITE in Wistar rats was established to be within the range of 0.5 – 1 mg/L.

 

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

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LC50
Value:
500 mg/m³
Quality of whole database:
One reliable key study is available for the acute inhalation toxicity of Lithium cryolite in the rat, performed according to the OECD Testing Guideline 403 and under GLP. The inhalatory LC50, 4h value in Wistar rats was established to be within the range of 0.5 – 1 mg/L.

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 12 September 2013 (Start of in-life phase) to 19 December 2013 (GLP compliance statement)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
not specified
Qualifier:
according to guideline
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. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approx. 10 weeks old)
- Weight at study initiation: mean weight for males was 288 g, mean weight for females was 199 g, Body weight variation did not exceed +/- 20% of the sex mean.
- Health inspection : At least prior to dosing. It was ensured that the animals were healthy and that the skin to be treated was intact and free from any abnormality
- Fasting period before study: none
- Housing: Individually housed in labeled Makrolon cages (MIII 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. During the acclimatization period the animals were group housed in Makrolon cages (MIV type, height 18 cm).

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): a 12-hour light/12-hour dark cycle

IN-LIFE DATES: From: 12 September 2013 To: 26 September 2013.
Type of coverage:
semiocclusive
Vehicle:
propylene glycol
Details on dermal exposure:
TEST SITE
- Clipping : One day before exposure (Day -1) an area of approximately 5x7 cm on the back of the animal was clipped.
- Area of exposure: approx. 25 cm² for males and 18 cm² for females
- % coverage: approx. 10% of the total body surface
- Type of wrap if used: The formulation was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done) :the skin cleaned of residual test substance using tap water.
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg (10 mL/kg) body weight

VEHICLE
Propylene glycol (Merck, Darmstadt, Germany) (specific gravity 1.036)
The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.
The formulation (w/w) was prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test substance.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 males and 5 females (females were nulliparous and non-pregnant).
Control animals:
not required
Details on study design:
- Duration of observation period following administration : 14 days
- Mortality/Viability : observation twice daily.
- Body weights : observation on days 1 (pre-administration), 8 and 15.
- Clinical signs : observations at periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).
- Necropsy : observation at the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.
Statistics:
Not applicable (limit test).
Preliminary study:
Not applicable.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Chromodacryorrhoea was noted for all animals on Days 1 and 2. General erythema, erythema maculate, scales and/or scabs were noted among the female animals between Days 3 and 15.
Body weight:
The changes noted in mean body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
None.

Deviations:

There were no deviations from the protocol.

Any deviations from standard operating procedures were evaluated and filed in the study file. There were no deviations from standard operating procedures that affected the integrity of the study.

Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD50 value of Lithium cryolite in Wistar rats was established to exceed 2000 mg/kg body weight.
Based on these results, Lithium cryolite does not have to be classified for acute dermal toxicity according to the CLP and the UN GHS.
Executive summary:

The acute dermal toxicity of Lithium cryolite in the rat was investigated according to the OECD Testing Guideline 402 and under GLP.

Lithium cryolite was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred.

Chromodacryorrhoea was noted for all animals on Days 1 and 2. General erythema, erythema maculate, scales and/or scabs were noted among the female animals between Days 3 and 15.

The mean body weight gain during the observation period was within the range expected for rats used in this type of study.

No abnormalities were found at macroscopic post mortem examination of the animals.

The dermal LD50 value of Lithium cryolite in Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results, Lithium cryolite does not have to be classified according to the CLP and the UN GHS.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Quality of whole database:
One reliable key study is available for the acute dermal toxicity of Lithium cryolite in the rat, performed according to the OECD Testing Guideline 402 and under GLP. No mortality occurred. The dermal LD50 value in Wistar rats was established to exceed 2000 mg/kg body weight.

Additional information

Justification for classification or non-classification

According to the OECD 423 test guideline, the oral LD50 value of Lithium cryolite in Wistar rats was established to be within the range of 300-2000 mg/kg body weight. Therefore, the substance should be classified as harmful if swallowed (Acute oral Tox. 4, H302) according to the CLP and the UN GHS.

 

Based on the OECD 403 test guideline, the inhalatory LC50, 4h value of LITHIUM CRYOLITE in Wistar rats was established to be within the range of 0.5 – 1 mg/L. Therefore, the substance should be classified as toxic if inhaled (Acute inhalation Tox. 3, H331) according to the CLP and the UN GHS.

 

Based on the OECD 402 test guideline, the dermal LD50 value of Lithium Cryolite in Wistar rats was established to exceed 2000 mg/kg body weight. Therefore, the substance does not have to be classified according to the CLP and the UN GHS for this route of exposure.