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Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 100 mg/L
Vehicle:
no
Test organisms (species):
Oryzias latipes
Details on test organisms:
TEST ORGANISM
- Common name: Japanese rice fish (Oryzias latipes)
- Age at study initiation: Test fish aged about 3-4 months
- Length at study initiation: Mean total length of 2.0 ± 1.0 cm
- Feeding during test: No feeding during test

ACCLIMATION
- Feeding: Fasted for 24 hours immediately prior to exposure


Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
10-250 mg/L CaCO3
Test temperature:
23.2 - 23.9 °C
pH:
7.13 - 7.97
Dissolved oxygen:
5.2 - 8.5 mg/L
Nominal and measured concentrations:
- nominal: 100 mg/L
- calculated test concentration based on measured calcium concentrations: 0.53 - 2.14 mg/L
Details on test conditions:
TEST SYSTEM
- Aeration: no aeration
- No. of organisms per concentration: 7 fish per test concentration without replication

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water was filtered to be used as culturing and dilution water. The tap water passed throug a membrane filter (1 µm) to remove particulate matter and through an activated carbon filter to remove the residual chlorine or organic substances.
- Residual Chlorine: below 0.01 mg/L

OTHER TEST CONDITIONS
- Photoperiod: 16 hours of light and 8 hours of dark

TEST CONCENTRATIONS
- Range finding study: The range-finding test was conducted at control, 0.1, 1, 10 and 100 mg/L and no mortality or adverse effects were observed at 100 mg/L.
- Test concentrations: 100 mg/L
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.
Remarks:
Ca3O8P2
Basis for effect:
mortality (fish)
Remarks on result:
other: measured concentration: >2.14 mg/L
Details on results:
- Mortality of control: non
- Effect concentrations exceeding solubility of substance in test medium: The concentrations of the test substance were calculated to be 0.53 - 2.14 mg/L based on measured calcium concentrations.
Reported statistics and error estimates:
Statistical analysis was not performed
Conclusions:
The 96 hours LC50 and NOEC of tricalcium phosphate were >100 mg/L (measured concentration: >2.14 mg/L) in O. latipes.
Executive summary:

A static freshwater toxicity test was conducted to determine the acute toxicity of tricalcium phosphate to Japanese rice fish, Oryzias latipes accorting to OECD Guideline 203. The concentration of the test substance were calculated to be 0.53 - 2.14 mg/L based on measured calcium concentrations. Therefore, the test substance is poorly water soluble. The results of the fish acute toxicity test showed that no mortality or adverse effects were observed at control or 100 mg/L (nominal). The 96 hours LC50 and no observed effect concentration (NOEC) in O. latipes were >100 mg/L (measured concentration: >2.14 mg/L).

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 100 mg/L
Vehicle:
no
Test organisms (species):
Oryzias latipes
Details on test organisms:
TEST ORGANISM
- Age at study initiation: Test fish aged about 3-4 months
- Length at study initiation: Mean total length of 2.0 ± 1.0 cm
- Feeding during test: No feeding during test

ACCLIMATION
- Feeding: Fasted for 24 hours immediately prior to exposure


Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
10-250 mg/L CaCO3
Test temperature:
23.2 - 23.9 °C
pH:
7.13 - 7.97
Dissolved oxygen:
5.2 - 8.5 mg/L
Nominal and measured concentrations:
- nominal: 100 mg/L
- calculated test concentration based on measured calcium concentrations: 13.5 mg/L
Details on test conditions:
TEST SYSTEM
- Aeration: no aeration
- No. of organisms per concentration: 7 fish per test concentration without replication

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water was filtered to be used as culturing and dilution water. The tap water passed throug a membrane filter (1 µm) to remove particulate matter and through an activated carbon filter to remove the residual chlorine or organic substances.
- Residual Chlorine: below 0.01 mg/L

OTHER TEST CONDITIONS
- Photoperiod: 16 hours of light and 8 hours of dark

TEST CONCENTRATIONS
- Range finding study: The range-finding test was conducted at control, 0.1, 1, 10 and 100 mg/L and no mortality or adverse effects were observed at 100 mg/L.
- Test concentrations: 100 mg/L
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.
Remarks:
CaHPO4
Basis for effect:
mortality (fish)
Remarks on result:
other: measured concentration: >13.5 mg/L
Details on results:
- Mortality of control: non
- Effect concentrations exceeding solubility of substance in test medium: The concentrations of the test substance were calculated to be 13.5 mg/L based on measured calcium concentrations.
Reported statistics and error estimates:
Statistical analysis was not performed
Conclusions:
The 96 hours LC50 and NOEC of tricalcium phosphate were >100 mg/L (measured concentration: >13.5 mg/L) in O. latipes.
Executive summary:

A static freshwater toxicity test was conducted to determine the acute toxicity of calcium hydrogen phosphate to Japanese rice fish, Oryzias latipes accorting to OECD Guideline 203. The concentration of the test substance were calculated to be 13.0 - 14.2 mg/L based on measured calcium concentrations. Therefore, the test substance is poorly water soluble. The results of the fish acute toxicity test showed that no mortality or adverse effects were observed at control or 100 mg/L (nominal). The 96 hours LC50 and no observed effect concentration (NOEC) in O. latipes were >100 mg/L (measured concentration: >13.5mg/L).

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1996-08-14 to 1996-12-6
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OTS 797.1400 (Fish Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Samples were analyzed for lithium in triplicate using a FLM-AAS method developed and validated by the FMC Corporation. Measured concentrations of lithium carbonate were calculated by multiplying measured lithium concentrations by 5.3240.
Vehicle:
no
Details on test solutions:
A primary stock solution (5001 mg/L) was prepared by adding 5.0007 g of neat test substance to a 1 L volumetric flask, bringing it to volume with deionized water and sonicating for approximately 50 minutes to ensure homogeneity. Test solutions were prepared by adding appropriate amounts of the primary stock to 9 L volumes of dilution water. A dilution water control was maintained concurrently with the solutions.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Rainbow trout used for the test ranged from 34 to 49 mm standard length (mean and standard deviation = 40 +/- 4.4 mm) and from 0.42 to 1.46 g wet weight (mean and standard deviation = 0.77 +/- 0.28 g) as measured from the control fish at test termination. Loading was calculated to be 0.86 g of fish tissue per liter of test solution.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
No
Hardness:
c(CaCO3)= 74 mg/L
Test temperature:
11.0 - 12.5 °C
pH:
The pH of the test solutions was affected by the presence of lithium carbonate (i.e., the pH increased as the test substance concentrations increased). The pH values of all test solutions ranged from 8.7 to 10.4 at test initiation and from 6.7 to 9.8 for the remainder of the test.
Dissolved oxygen:
11.7 - 12.2 mg/L (>100 % of saturation)
Nominal and measured concentrations:
Nominal concentrations: 5.0, 10, 20, 40, and 80 mg/L
Measured concentrations: 0.0, 4.99, 10.0, 19.1, 38.9 and 77.7 mg/L
Details on test conditions:
A 96-hour static range-finding test was conducted at nominal concentrations of 0.10, 1.00, 10.0, 100, and 1000 mg/L, prior to performing the definitive test. Three fish were tested at each concentration. After 96 hours of exposure, mortality of rainbow trout in the range-finding test was 0 % at the test concentrations 0.10, 1.00, and 10.0 mg/L, and 100 % at 100 and 1000 mg/L. Mortality in the control was 0 %. Undissolved compound was observed at 100 mg/L during the range-finding test. Based upon these results, nominal test concentrations of 5.0, 10, 20, 40, and 80 mg/L were selected for the definitive test.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
30.3 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Remarks:
Li2CO3
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% CL: 19.1 and 38.9 mg wm/L
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
19.1 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Remarks:
Li2CO3
Basis for effect:
mortality (fish)
Details on results:
The 96-hour LC50 was 30.3 mg/L (based upon mean measured concentrations) with 95 % confidence limits of 19.1 and 38.9 mg/L. The no-observed-effect-concentration (NOEC) was 19.1 mg/L based on the lack of significant mortality and the absence of sublethal effects at this and lower test concentrations.
Reported statistics and error estimates:
The LC50 values were estimated by a computer program using the following statistical methods: moving average angle, probit, logit, and non-linear interpolation. Confidence limits for LC50 values determined by non-linear interpolation were calculated by binomial probability. The method selected for reporting the test results was determined by the characteristics of the data, i.e. the presence or absence of 0 % and 100 % mortality and the number of concentrations in which mortalities between 0 and 100 % occurred.
Sublethal observations / clinical signs:

Mortality of Rainbow Trout, Oncorhynchus mykiss, during a 96-hour static exposure to lithium carbonate:

Mean Measured Concentration
(mg/L)

Cumulative Number Dead (Percent Mortality)

24 Hour

48 Hour

72 Hour

96 Hour

Control

0

(0)

0

(0)

0

(0)

0

(0)

4.99

0

(0)

0

(0)

0

(0)

0

(0)

10.0

0

(0)

0

(0)

0

(0)

0

(0)

19.1

0

(0)

0

(0)

0

(0)

0

(0)

38.9

0a

(0)

5

(25)

15

(75)

17

(85)

77.7

8b

(40)

19

(95)

20

(100)

20

(100)

a Two fish exhibited dark coloration.

b Five fish exhibited dark coloration.

 

Calculated LC50 values, based upon measured concentrations for Rainbow Trout, Oncorhynchus mykiss, exposed to lithium carbonate under static test conditions:

Exposure Period (Hour)

LC50 Value a

LCL b

UCL c

Slope

Method Used

24

>77.7

-

-

-

-

48

47.6

40.5

56.4

7.81

Probit

72

32.1

19.1

38.9

N/A

Binomial

96

30.3

19.1

38.9

N/A

Binomial

a LC50 values and confidence limits expressed in mg wm/L

b Lower 95 percent confidence limit

c Upper 95 percent confidence limit

Validity criteria fulfilled:
yes
Conclusions:
The 96-hour LC50 was 30.3 mg lithium carbonate/L (based upon mean measured concentrations) with 95 % confidence limits of 19.1 and 38.9 mg/L. The NOEC was 19.1 mg lithium carbonate/L based on the lack of significant mortality at this and lower test concentrations.
Executive summary:

A static freshwater toxicity test was conducted to determine the acute toxicity of lithium carbonate to rainbow trout, Oncorhynchus mykiss according to OECD Guideline 203. Mean measured concentration of lithium carbonate ranged from 4.99 to 77.7 mg /L and from 96 to 100 % of nominal. All test solutions appeared clear and colourless and concentrations remained stable throughout the test. The pH of the test solutions was affected by the presence of lithium carbonate (i.e. the pH increased as the test substance concentrations increased). The pH values of all test solutions ranged from 8.7 to 10.4 at test initiation and from 6.7 to 9.8 for the remainder of the test. Mortality of the rainbow trout exposed for 96 hours to lithium carbonate ranged from 0 % at test concentrations 19.1 mg /L to 100 % at 77.7 mg /L. No mortality occurred in the dilution water control. The 96-hour LC50 was 30.3 mg lithium carbonate/L with 95 % confidence limits of 19.1 and 38.9 mg/L. The NOEC of 19.1 mg lithium carbonate/L was based on a lack of significant mortality and the absence of sublethal effects at this and all lower test concentrations.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1997-06-13 to 1997-12-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1992
Deviations:
no
Principles of method if other than guideline:
no
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch No.of test material: 410c
Analytical monitoring:
yes
Details on sampling:
Samples were analyzed for lithium in triplicate using a FLM-AAS method developed and validated by the FMC Corporation. Measured concentrations of lithium chloride were calculated by multiplying measured lithium concentrations by 14.427.
Vehicle:
no
Details on test solutions:
Test solutions were prepared by adding appropriate amounts of lithium chloride (0.56, 1.12, 2.25, 4.50 and 9.00 g) to 9 L volumes of dilution water. The A and B replicates were prepared separately. A control of only dilution water was maintained concurrently with the test solutions.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Rainbow trout used for the test ranged from 24 to 30 mm standard length (mean and standard deviation = 27 +/- 1.6 mm) and from 0.13 to 0.27 g wet weight (mean and standard deviation = 0.2 +/- 0.05 g) as measured from the control fish at test termination. Loading was calculated to be 0.22 g of fish tissue per liter of test solution.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
c(CaCO3)= 56 mg/L
Test temperature:
11.9 - 13.4 °C
pH:
The pH of the test solutions was affected by the presence of lithium chloride (i.e., the pH increased as the test substance concentrations increased).
The pH values of all test solutions ranged from 7.5 to 8.7 at test initiation and from 7.0 to 7.5 for the remainder of the test.
Dissolved oxygen:
10.6 to 11.0 mg/L (98 - 102 % of saturation) at test initiation;
≥6.6 mg/L (≥61 % of saturation) for the remainder of the test
Salinity:
Not applicable
Nominal and measured concentrations:
Nominal concentrations: Control, 62.5, 125, 250, 500 and 1000 mg LiCl/L
Measured concentrations: 0.0, 59.4, 188, 249, 516 and 1021 mg LiCl/L
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
158 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Remarks:
LiCl
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% CL: 118 and 249 mg/L
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
59.4 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Remarks:
LiCl
Basis for effect:
mortality (fish)
Details on results:
The 96-hour LC50 was 158 mg/L (based upon mean measured concentrations) with 95 % confidence limits of 118 and 249 mg/L. The NOEC was 59.4 mg/L based on the lack of mortality at this, the lowest, test concentration.
Results with reference substance (positive control):
No reference substance control
Reported statistics and error estimates:
The LC50 values were estimated by a computer program using the following statistical methods: moving average angle, probit, logit, and non-linear interpolation. Confidence limits for LC50 values determined by non-linear interpolation were calculated by binomial probability. The method selected for reporting the test results was determined by the characteristics of the data, i.e., the presence or absence of 0 % and 100 % mortality and the number of concentrations in which mortalities between 0 and 100 % occurred.
Validity criteria fulfilled:
yes
Conclusions:
The 96 hour LC50 was 158 mg/L (based upon mean measured concentrations) with 95 % confidence limits of 118 and 249 mg/L. The NOEC was 59.4 mg/L based on the lack of mortality at this, the lowest, test concentration.
Executive summary:

A static freshwater toxicity test was conducted to determine the acute toxicity of lithium chloride to rainbow trout, Oncorhynchus mykiss according to OECD Guideline No. 203. Mean measured concentration of lithium chloride ranged from 59.4 to 1021 mg/L and from 94 to 103 % of nominal concentrations. All test solutions appeared clear and colourless and concentrations remained stable throughout the test. The pH of the test solutions was affected by the presence of lithium chloride (i.e., the pH increased as the test substance concentrations increased). The pH values of all test solutions ranged from 7.5 to 8.7 at test initiation and from 7.0 to 7.5 for the remainder of the test. Mortality of the rainbow trout exposed for 96 hours to lithium chloride ranged from 0 % at 59.4 mg/L to 100 % at test concentrations smaller or equal to 249 mg/L. No mortality occurred in the dilution water control. The 96-hour LC50 was 158 mg/L with 95 % confidence limits of 118 and 249 mg/L. The slope of the concentration response curve could not be calculated using the binomial method. The NOEC was 59.4 mg/L based on a lack of mortality at the lowest test concentration.

Description of key information

The 96-hour LC50 was 30.3 mg lithium carbonate/L (based upon mean measured concentrations) with 95 % confidence limits of 19.1 and 38.9 mg/L. The NOEC was 19.1 mg lithium carbonate/L based on the lack of significant mortality at this and lower test concentrations. Based on read-across approach, the calculated 96-hour LC50 for lithium phosphate is 31.65 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
31.65 mg/L

Additional information

A short-term toxicity study on fish with lithium phosphate is not available. Consequently, read-across was applied using characteristically similar compounds: lithium carbonate and chloride as well as tricalcium phosphate and calcium hydrogen phosphate.

Read-across with lithium carbonate (Toxikon 1996)
A static freshwater toxicity test was conducted to determine the acute toxicity of lithium carbonate to rainbow trout, Oncorhynchus mykiss according to OECD Guideline 203. Mean measured concentration of lithium carbonate ranged from 4.99 to 77.7 mg/L and from 96 to 100 % of nominal. All test solutions appeared clear and colourless and concentrations remained stable throughout the test. The pH of the test solutions was affected by the presence of lithium carbonate (i.e. the pH increased as the test substance concentrations increased). The pH values of all test solutions ranged from 8.7 to 10.4 at test initiation and from 6.7 to 9.8 for the remainder of the test. Mortality of the rainbow trout exposed for 96 hours to lithium carbonate ranged from 0 % at test concentrations 19.1 mg/L to 100 % at 77.7 mg/L. No mortality occurred in the dilution water control. The 96-hour LC50 was 30.3 mg Li2CO3/L with 95 % confidence limits of 19.1 and 38.9 mg/L. The NOEC of 19.1 mg Li2CO3/L was based on a lack of significant mortality and the absence of sublethal effects at this and all lower test concentrations. Based on these data, the calculated LC50 for lithium phosphate is 31.65 mg/L.

Read-across with lithium chloride (Toxicon 1997) 
A static freshwater toxicity test was conducted to determine the acute toxicity of lithium chloride to rainbow trout, Oncorhynchus mykiss according to OECD Guideline No. 203. Mean measured concentration of lithium chloride ranged from 59.4 to 1021 mg/L and from 94 to 103 % of nominal concentrations. All test solutions appeared clear and colourless and concentrations remained stable throughout the test. The pH of the test solutions was affected by the presence of lithium chloride (i.e., the pH increased as the test substance concentrations increased). The pH values of all test solutions ranged from 7.5 to 8.7 at test initiation and from 7.0 to 7.5 for the remainder of the test. Mortality of the rainbow trout exposed for 96 hours to lithium chloride ranged from 0 % at 59.4 mg/L to 100 % at test concentrations smaller or equal to 249 mg/L. No mortality occurred in the dilution water control. The 96-hour LC50 was 158 mg/L with 95 % confidence limits of 118 and 249 mg/L. The slope of the concentration response curve could not be calculated using the binomial method. The NOEC was 59.4 mg/L based on a lack of mortality at the lowest test concentration. Based on these data, the calculated LC50 for lithium phosphate is 143.86 mg/L. 

Read-across with tricalcium phosphate (Kim et al. 2013) 
A static freshwater toxicity test was conducted to determine the acute toxicity of tricalcium phosphate to Japanese rice fish, Oryzias latipes according to OECD Guideline 203. The concentrations of the poorly soluble test substance were calculated to be 0.53 -2.14 mg/L based on measured calcium concentrations. The results of the fish acute toxicity test showed that no mortality or adverse effects were observed at control or 100 mg/L (nominal). The 96 hours LC50 and no observed effect concentration (NOEC) in O. latipes were >100 mg/L (measured concentration: >2.14 mg/L). Based on these data, the calculated LC50 for lithium phosphate is >74.66 mg/L.

Read-across with calcium hydrogen phosphate (Kim et al. 2013) 
A static freshwater toxicity test was conducted to determine the acute toxicity of calcium hydrogen phosphate to Japanese rice fish, Oryzias latipes according to OECD Guideline 203. The concentrations of the test substance were calculated to be 13.0 - 14.2 mg/L based on measured calcium concentrations. Therefore, the test substance is poorly water soluble. The results of the fish acute toxicity test showed that no mortality or adverse effects were observed at control or 100 mg/L (nominal). The 96 hours LC50 and no observed effect concentration (NOEC) in O. latipes were >100 mg/L (measured concentration: >13.5 mg/L). Based on the read-across approach, the calculated LC50 for lithium phosphate is >85.10 mg/L. 

Conclusion:

In this weight of evidence approach the effect concentrations of lithium and phosphate compounds were evaluated in different fish tests. When comparing the results of read-across from lithium carbonate and lithium chloride with tricalcium phosphate and calcium hydrogen phosphate, the magnitude of the calculated LC50 values show that lithium is the relevant toxicological moiety of lithium phosphate with respect to acute toxicity in fish. The result of the acute fish test with lithium carbonate with the guideline compliant species O. mykiss is considered as valid and the LC50 of 31.65 mg Li3PO4/L is chosen as key value for chemical safety assessment.