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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Experimental result using OECD recommended guidelines
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
no
Analytical monitoring:
yes
Vehicle:
not specified
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solution was prepared by dissolving 500 mg of test chemical in 5000 ml of RO water to get the final concentration of 100 mg/L. The remaining test solutions were prepared by dilution from the above stock solution.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra fish
- Source: Anu pet shop, Nagpur
- Length at study initiation (length definition, mean, range and SD): 1.96 cm
- Housing: The fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration.
- Duration of housing: 7 days

ACCLIMATION
- Acclimation period: Fishes, to be used in this study, are kept in the test water for seven days immediately before testing
- Acclimation conditions (same as test or not): same test conditions
- Photoperiod: 16 hour light and 8 hours dark
- Temperature range 23.17°C
- pH: 7.31
- DO: 8.03 mg/L (97.225%)
- Feeding: Daily once but not given atleast 48 hours before study
- Aeration: Continuous
- Diet: Standard brand feed, once daily
- Health during acclimation (any mortality observed): no

FEEDING DURING TEST (as applicable)
- Food type: Feeding was not provided during test.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
125 mg/l as CaCO3
Test temperature:
18 to 22 °C
pH:
7.3
Dissolved oxygen:
7.1 to 8.2 mg/l
Nominal and measured concentrations:
0 (control), 10, 11, 12, 19.2, 30.72, 49.152 and 78.643 mg/l (nominal conc.)
Details on test conditions:
TEST SYSTEM
- Test vessel: glass aquaria of 7 lit capacity
- Material, size, headspace, fill volume: 4000 mL for 12, 19.2, 30.72, 49.152, 78.643 mg/L whereas 2500 ml for 10 and 11 mg/L
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates):1
- No. of vessels per control (replicates):1

OTHER TEST CONDITIONS
- Photoperiod: 16:8 hrs (light:dark)
- Light intensity: 520 to 980 lux

TEST CONCENTRATIONS
- Spacing factor for test concentrations: <2.2
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
10.25 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: details not known
Sublethal observations / clinical signs:

Table: pH, TEMPERATURE, AND DISSOLVE OXYGEN


 








































































Test Concentration(mg/L)



pH



24 Hours



48 Hours



72 Hours



96 Hours



control



7.1



7.1



7.5



6.9



10



7.4



7.3



7.7



7.1



11



7.5



7.3



7.6



7.3



12



-



-



-



-



19.2



-



-



-



-



30.72



-



-



-



-



49.152



-



-



-



-



78.643



-



-



-



-



 








































































Test Concentration(mg/L)



Temperature °C



24 Hours



48 Hours



72 Hours



96 Hours



control



23.2



23.3



23.2



23.2



10



23.2



23.3



23.2



23.2



11



23.2



23.2



23.2



23.2



12



-



-



-



-



19.2



-



-



-



-



30.72



-



-



-



-



49.152



-



-



-



-



78.643



-



-



-



-



 








































































Test Concentration(mg/L)



Dissolved Oxygen



24 Hours



48 Hours



72 Hours



96 Hours



control



101.1



101.3



93.5



83.8



10



98.2



96.4



94.7



83.3



11



101.4



99.2



95.5



89.7



12



-



-



-



-



19.2



-



-



-



-



30.72



-



-



-



-



49.152



-



-



-



-



78.643



-



-



-



-


Validity criteria fulfilled:
yes
Conclusions:
Based on the effect on toxicity to fish, the 96 hr LC50 value of test chemical was determined to be 10.25 mg/L.
Executive summary:

A short-term toxicity test was performed as per OECD Guideline 203 (Fish, Acute toxicity test) assesses the acute toxic effects (mortality) of various concentrations of a test chemical on a freshwater fish species.  Danio rerio (zebrafish) of length 1.96 cm was used as a test organism under static conditions for 96 hrs of exposure. A group of 7 fish of recommended species was exposed for a period of 96 hrs. The solution was prepared by dissolving 500 mg of the test chemical in 5000 ml of RO water to get the final concentration of 100 mg/L. The remaining test solutions were prepared by dilution from the above stock solution. 7 test concentrations control, 10, 11, 12,19.2,30.72,49.152,78.643 mg/L of test chemical dissolved in 5000 ml of water. The auxiliaries used to prepare the test media were magnetic stirrers. The analytical determinations were performed by UV Spectrophotometer and HPLC. Analytical assessments were performed for selected test concentrations at 0 Hr and 96 Hr. On condition of ACN (100%) (1 ml/min), 25°C with column C-18. For concentrations 12, 19.2, 30.72, 49.152, and 78.643 mg/L analytical were performed by using a UV Spectrophotometer. And for concentrations 10 and 11 mg/L analytical was performed by using HPLC as these lower concentrations were not detected on UV Spectrophotometer because of its limit of detection (LOD is 1 mg/l and LOQ is 10 mg/l). The reproducibility range was between 80-120% of the exposure test chemical. The test substance was identified based on the absorbance at zero-day and 96 hours. The concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test so, analysis of the results was based on nominal concentration. The test concentrations were measured and found to remain at 120-80% of the nominal. Hence, the LC50 was expressed based on Nominal concentrations and the spacing factor for test concentrations was <2.2. The fish were maintained and kept in a static tank in tap water passed through a reverse osmosis system, under natural conditions along with proper feed and aeration for 7 days. Fishes are kept in the test water for seven days immediately before testing under same condition as like test conditions. Fishes, to be used in this study, are kept for acclimatization in the test water for seven days immediately before testing in the same test conditions. Fishes were fed by standard brand feed daily once but not given at least 48 hours before the study. No mortalities were found during the period of acclimatization. Test condition involves photoperiod 16-hour light and 8 hours dark, temperature range 18 to 22 °C, pH 7.31, DO 7.1-8.2 mg/L, hardness 125 mg/l as CaCO3 respectively. Continuous aeration was provided. Glass aquaria of 7 lit capacity filled with volume 4000 mL for 12, 19.2, 30.72, 49.152, 78.643 mg/L whereas 2500 ml for 10 and 11 mg/L, respectively. No. of organisms per vessel 7 with 1 replicate. The mortality in the controls did not exceed 10% by the end of the test and dissolved oxygen concentration remained above 60% of the air-saturation value throughout the exposure period. Hence, the test fulfilled the validity criteria as per OECD 203. Based on the effect on toxicity to fish, the 96 hr LC50 value of the test chemical was determined to be 10.25 mg/L. Since the test chemical was readily biodegradable, hence the test chemical was not classified as per CLP classification criteria.


 


                                                                                         


 

Description of key information

Short-term toxicity to fish:


A short-term toxicity test was performed as per OECD Guideline 203 (Fish, Acute toxicity test) assesses the acute toxic effects (mortality) of various concentrations of a test chemical on a freshwater fish species.  Danio rerio (zebrafish) of length 1.96 cm was used as a test organism under static conditions for 96 hrs of exposure. A group of 7 fish of recommended species was exposed for a period of 96 hrs. The solution was prepared by dissolving 500 mg of the test chemical in 5000 ml of RO water to get the final concentration of 100 mg/L. The remaining test solutions were prepared by dilution from the above stock solution. 7 test concentrations control, 10, 11, 12,19.2,30.72,49.152,78.643 mg/L of test chemical dissolved in 5000 ml of water. The auxiliaries used to prepare the test media were magnetic stirrers. The analytical determinations were performed by UV Spectrophotometer and HPLC. Analytical assessments were performed for selected test concentrations at 0 Hr and 96 Hr. On condition of ACN (100%) (1 ml/min), 25°C with column C-18. For concentrations 12, 19.2, 30.72, 49.152, and 78.643 mg/L analytical were performed by using a UV Spectrophotometer. And for concentrations 10 and 11 mg/L analytical was performed by using HPLC as these lower concentrations were not detected on UV Spectrophotometer because of its limit of detection (LOD is 1 mg/l and LOQ is 10 mg/l). The reproducibility range was between 80-120% of the exposure test chemical. The test substance was identified based on the absorbance at zero-day and 96 hours. The concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test so, analysis of the results was based on nominal concentration. The test concentrations were measured and found to remain at 120-80% of the nominal. Hence, the LC50 was expressed based on Nominal concentrations and the spacing factor for test concentrations was <2.2. The fish were maintained and kept in a static tank in tap water passed through a reverse osmosis system, under natural conditions along with proper feed and aeration for 7 days. Fishes are kept in the test water for seven days immediately before testing under same condition as like test conditions. Fishes, to be used in this study, are kept for acclimatization in the test water for seven days immediately before testing in the same test conditions. Fishes were fed by standard brand feed daily once but not given at least 48 hours before the study. No mortalities were found during the period of acclimatization. Test condition involves photoperiod 16-hour light and 8 hours dark, temperature range 18 to 22 °C, pH 7.31, DO 7.1-8.2 mg/L, hardness 125 mg/l as CaCO3 respectively. Continuous aeration was provided. Glass aquaria of 7 lit capacity filled with volume 4000 mL for 12, 19.2, 30.72, 49.152, 78.643 mg/L whereas 2500 ml for 10 and 11 mg/L, respectively. No. of organisms per vessel 7 with 1 replicate. The mortality in the controls did not exceed 10% by the end of the test and dissolved oxygen concentration remained above 60% of the air-saturation value throughout the exposure period. Hence, the test fulfilled the validity criteria as per OECD 203. Based on the effect on toxicity to fish, the 96 hr LC50 value of the test chemical was determined to be 10.25 mg/L. Since the test chemical was readily biodegradable, hence the test chemical was not classified as per CLP classification criteria.


 


                                                                                         


 


 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Dose descriptor:
LC50
Effect concentration:
10.25 mg/L

Additional information

Short-term toxicity to fish:


 


A short-term toxicity test was performed as per OECD Guideline 203 (Fish, Acute toxicity test) assesses the acute toxic effects (mortality) of various concentrations of a test chemical on a freshwater fish species.  Danio rerio (zebrafish) of length 1.96 cm was used as a test organism under static conditions for 96 hrs of exposure. A group of 7 fish of recommended species was exposed for a period of 96 hrs. The solution was prepared by dissolving 500 mg of the test chemical in 5000 ml of RO water to get the final concentration of 100 mg/L. The remaining test solutions were prepared by dilution from the above stock solution. 7 test concentrations control, 10, 11, 12,19.2,30.72,49.152,78.643 mg/L of test chemical dissolved in 5000 ml of water. The auxiliaries used to prepare the test media were magnetic stirrers. The analytical determinations were performed by UV Spectrophotometer and HPLC. Analytical assessments were performed for selected test concentrations at 0 Hr and 96 Hr. On condition of ACN (100%) (1 ml/min), 25°C with column C-18. For concentrations 12, 19.2, 30.72, 49.152, and 78.643 mg/L analytical were performed by using a UV Spectrophotometer. And for concentrations 10 and 11 mg/L analytical was performed by using HPLC as these lower concentrations were not detected on UV Spectrophotometer because of its limit of detection (LOD is 1 mg/l and LOQ is 10 mg/l). The reproducibility range was between 80-120% of the exposure test chemical. The test substance was identified based on the absorbance at zero-day and 96 hours. The concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test so, analysis of the results was based on nominal concentration. The test concentrations were measured and found to remain at 120-80% of the nominal. Hence, the LC50 was expressed based on Nominal concentrations and the spacing factor for test concentrations was <2.2. The fish were maintained and kept in a static tank in tap water passed through a reverse osmosis system, under natural conditions along with proper feed and aeration for 7 days. Fishes are kept in the test water for seven days immediately before testing under same condition as like test conditions. Fishes, to be used in this study, are kept for acclimatization in the test water for seven days immediately before testing in the same test conditions. Fishes were fed by standard brand feed daily once but not given at least 48 hours before the study. No mortalities were found during the period of acclimatization. Test condition involves photoperiod 16-hour light and 8 hours dark, temperature range 18 to 22 °C, pH 7.31, DO 7.1-8.2 mg/L, hardness 125 mg/l as CaCO3 respectively. Continuous aeration was provided. Glass aquaria of 7 lit capacity filled with volume 4000 mL for 12, 19.2, 30.72, 49.152, 78.643 mg/L whereas 2500 ml for 10 and 11 mg/L, respectively. No. of organisms per vessel 7 with 1 replicate. The mortality in the controls did not exceed 10% by the end of the test and dissolved oxygen concentration remained above 60% of the air-saturation value throughout the exposure period. Hence, the test fulfilled the validity criteria as per OECD 203. Based on the effect on toxicity to fish, the 96 hr LC50 value of the test chemical was determined to be 10.25 mg/L. Since the test chemical was readily biodegradable, hence the test chemical was not classified as per CLP classification criteria.