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Ecotoxicological information

Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Study initiation date : 31/10/2013, study completion date : 12/05/2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
The test samples were taken from test water of fish acute toxicity test.
Vehicle:
no
Details on test solutions:
Range-finding test :
During the pre-test a white precipitation was observed in the test solutions of the treatment groups. The test substance (molecular formula: Li3AlF6) contains aluminum (Al) element, so the precipitation could result from the formation of low soluble salts, i.e., the reaction product between aluminum and anions present in the test water. Thus, a 24h semi-static test was performed to maintain the concentration of the test substance as far as possible in the range-finding test and limit test. The test solution of each treatment was prepared at its respective nominal concentration: a certain amount of the test substance was added into 5L test water and stirred (via magnetic stirrer) in the dark for 72h at 500rpm followed by filtration using a 0.45μm nitrocellulose membrane. After preparation the test solutions were clear and no precipitation was visible. Before the initiation of the test, pH values of the respective test solution were adjusted approximately to 7.0 by 1M HCl

Limit test :
A 24h semi-static test was performed to maintain the concentration of the test substance as far as possible in the limit test. A nominal concentration of 100mg/L of the test substance was prepared and stirred (via magnetic stirrer) in the dark for 72h at 500rpm followed by filtration using a 0.45μm nitrocellulose membrane as the test solution. After preparation the test solutions were clear and no precipitation was visible. Before the initiation of the test, pH values of the respective test solution were adjusted approximately to 7.0 by 1M HCl.
Test organisms (species):
other: Gobiocypris rarus
Details on test organisms:
The test was performed with Gobiocypris rarus which were bred in Bioassay and Safety Assessment Lab, Shanghai Academy of Public Measurement. This freshwater fish species is suitable for fish toxicity testing because of their ease of culturing and handling, sensitivity to a variety of chemical substances, and the extensive data base for this species. This species complies with OECD Guidelines for the Testing of Chemicals. 203 Fish, Acute Toxicity Test 1992.
Before being used in the test, all fish were acclimatized in the following test conditions for more than 14 days:
Water: the test water used was tap water dechlorinated by activated carbon and sterilized by UV lamp;
Temperature: 21-25℃ and constant within a range of 2℃.
pH: 6.0-8.5;
Dissolved oxygen concentration: > 80% of air saturation value (ASV);
Photoperiod: Light∶Dark = 12 hours∶12 hours;
Feeding: fish were fed daily until 24 hours before the test was started.
Length was 2.6±0.1 cm (Mean ± SD) and weight was 0.33±0.03g (Mean ± SD) at the end of the test.
No diseases were observed during the acclimation period;
No mortality was present within the 7 days before the start of the test.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
Total hardness: 155mg CaCO3/L
Test temperature:
Measured from 22.1 to 23.0℃ during the limit test.
pH:
Measured from 6.64 to 6.99 during the limit test.
Dissolved oxygen:
Measured from 75.8 to 92.8 % ASV during the limit test.
Salinity:
No data
Nominal and measured concentrations:
Range-finding test: Two nominal concentrations of 12.0mg/L and 100mg/L.
Limit test: Nominal concentration of 100 mg/L, the geometric mean measured concentrations of F and Al were 52.3mg/L and 8.55mg/L, respectively.
Details on test conditions:
Water used in the test : Tap water, dechlorinated by activated carbon and sterilized by UV lamp, was used. The water was aerated continuously over 24h. Water quality parameters were as follows:
Total hardness: 155mg CaCO3/L;
Dissolved oxygen concentration: >90% of ASV.

Test vessels: 5L glass tanks were used containing 4L test solution.

Conditions of exposure:
- Duration: 96h;
- Test method: semi-static and the frequency of renewal was every 24h;
- Loading: about 0.8g fish/litre;
- Temperature: 22.1-23.0℃;
- Photoperiod: Light∶Dark=12 hours∶12 hours;
- Dissolved oxygen concentration: 75.8 -92.8%ASV;
- Feeding: none.

Procedure:
- Range-finding test :
In this test, a blank control of test water was used. At each renewal, fresh test solutions were used. Five fish were introduced into each concentration and the test duration was 96 hours. Fish were observed and dead fish were recorded at 0, 24, 48, 72 and 96 hours after test initiation. For the analysis of the concentrations of F and Al, two samples of 10mL test solution were taken from each test vessel at the beginning, before and after each renewal, and at the end of the test. In the range-finding test, there were no mortalities and abnormalities observed in the fish of the control group and the treatment groups during the test
- Limit test :
Based on the results of the range-finding test, a limit test was performed. In this test, a blank control of test water and a nominal concentration of 100mg/L were used. At each renewal, fresh test solutions were used.Ten fish were introduced into each concentration and the test duration was 96 hours.
The fish were inspected at 0, 24, 48, 72 and 96 hours after test initiation and mortalities and abnormalities were recorded. Measurements of the pH, dissolved oxygen concentration and temperature in the test solutions were made at the beginning, before and after each renewal and at the end of the test. For the analysis of the concentrations of F and Al, two samples of 10mL test solution were taken from each test vessel at the beginning, before and after each renewal, and at the end of the test.
At the end of the test, the body length and body weight of the test fish of the control group were measured. The loading rate was also calculated with the body weight at the same time.


Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
Analytical measurements in the limit test:
The measured concentrations of F ranged from 47.8 to 57 mg/L. The geometric mean measured concentration was 52.3 mg/l with a variation of less than 20% (+/- 8.96%).
The measured concentrations of Al ranged from 7.22 to 9.46 mg/L. The geometric mean measured concentration was 8.55 mg/l with a variation of less than 20% (+/- 18.1%).
Since the test substance is a metal compound consisting of several elements (Li, Al and F), the results were expressed as the nominal concentrations in order to reflect the toxicity of the test substance.

There were no mortalities and abnormalities observed in the fish of the control group and the treatment group during the test.

Validity criteria:
- No mortality was observed in the control during the test;
- Acceptable exposure conditions were maintained throughout the test;
- The dissolved oxygen concentrations in test solutions were between 75.8-92.8%ASV, which was greater than 60.0% of the air saturation value, throughout the test.
- During the test, the measured concentrations of both F and Al varied less than 20% of their geometric mean measured concentrations.
The test fulfilled the above requirements and was therefore considered to be valid.

Results with reference substance (positive control):
No data
Reported statistics and error estimates:
Not applicable
Validity criteria fulfilled:
yes
Conclusions:
The test results showed that the 96h LC50 for the test substance to Gobiocypris rarus was higher than 100mg/L.
Executive summary:

The acute toxicity of Lithium cryolite to Gobiocypris rarus was determined in a semi-static test according to the OECD Testing Guideline 2013 and under GLP.

During the pre-test, a white precipitation was observed in the test solutions of the treatment groups. The test substance contains aluminum (Al) element, so the precipitation could result from the formation of low soluble salts, i.e., the reaction product between aluminum and anions (OH-) present in the test water. Thus, a 24h semi-static test was performed to maintain the concentration of the test substance as far as possible in the range-finding test and limit test. The test solution of each treatment was prepared at its respective nominal concentration: a certain amount of the test substance was added into 5L test water and stirred (via magnetic stirrer) in the dark for 72h at 500rpm followed by filtration using a 0.45μm nitrocellulose membrane. Based on the results of the range-finding test, a nominal concentration of 100mg/L and a blank control were used to perform the limit test. The treatment group and control group had one replicate, each containing 10 fish and the test duration was 96h.

There were no mortalities and abnormalities observed in treatment group during the test period.

No mortality occurred in the control during the test and the dissolved oxygen concentration was greater than 60.0% of the air saturation value (ASV) throughout the test duration. Hence, the test was considered to be valid.

During the test, the measured concentrations of fluoride (F) analysed by ion chromatography (IC) varied by -8.69%-8.96%, which was less than 20% of the geometric mean measured concentrations. The measured concentration of aluminum (Al) analysed by ICP-MS, varied by -15.6%-18.1%, which was less than 20% of the geometric mean measured concentrations.

Since the test substance is a metal compound consisting of several elements (Li, Al and F), the results were expressed as the nominal concentrations in order to reflect the toxicity of the test substance.

The 96h LC50 for the test substance to Gobiocypris rarus was higher than 100mg/L (the geometric mean measured concentrations of F and Al were 52.3mg/L and 8.55mg/L, respectively) under tested conditions.

Description of key information

A reliable acute limit toxicity test is available, performed under semi-static conditions.

No mortality was observed during the test and therefore, the 96h LC50 was determined to be above 100 mg/L.

Key value for chemical safety assessment

Additional information

One reliable key study is available for this endpoint. In this study, the acute toxicity of Lithium cryolite to Gobiocypris rarus was determined in a semi-static conditions according to the OECD Testing Guideline 2013 and under GLP.

During the pre-test, a white precipitation was observed in the test solutions of the treatment groups. The test substance contains aluminum (Al) element, so the precipitation could result from the formation of low soluble salts, i.e., the reaction product between aluminum and anions (OH-) present in the test water. Thus, a 24h semi-static test was performed to maintain the concentration of the test substance as far as possible in the range-finding test and limit test. The test solution of each treatment was prepared at its respective nominal concentration: a certain amount of the test substance was added into 5L test water and stirred (via magnetic stirrer) in the dark for 72h at 500rpm followed by filtration using a 0.45μm nitrocellulose membrane. Based on the results of the range-finding test, a nominal concentration of 100mg/L and a blank control were used to perform the limit test. The treatment group and control group had one replicate, each containing 10 fish and the test duration was 96h.

There were no mortalities and abnormalities observed in treatment group during the test period.

No mortality occurred in the control during the test and the dissolved oxygen concentration was greater than 60.0% of the air saturation value (ASV) throughout the test duration. Hence, the test was considered to be valid.

During the test, the measured concentrations of fluoride (F) analysed by ion chromatography (IC) varied by -8.69%-8.96%, which was less than 20% of the geometric mean measured concentrations. The measured concentration of aluminum (Al) analysed by ICP-MS, varied by -15.6%-18.1%, which was less than 20% of the geometric mean measured concentrations.

Since the test substance is a metal compound consisting of several elements (Li, Al and F), the results were expressed as the nominal concentrations in order to reflect the toxicity of the test substance.

The 96h LC50 for the test substance to Gobiocypris rarus was higher than 100mg/L (the geometric mean measured concentrations of F and Al were 52.3mg/L and 8.55mg/L, respectively) under tested conditions.