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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Experimental test result performed using standard OECD test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
The stock solution 100 mg/l was prepared by dissolving test chemical in reconstituted water. The test solutions of required concentrations were prepared by mixing the stock solution of the test sample in reconstituted water.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Strain: Straus
- Source: Own breeding at University of Chemistry and Technology, Prague
- Age at study initiation (mean and range, SD): The animals used for the test shall be less than 24 h old and should not be first brood progeny
- Feeding during test: No feeding
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
± 1 hr
Test temperature:
20±1°C
pH:
without adjustment
sample at concentration 80 mg/l: pH = 7.8 changed to pH = 7.9 during the test
control: pH = 7.7 did not change during the test
Dissolved oxygen:
higher than 7.7 mg/L at the end of test both in the control and the sample
Nominal and measured concentrations:
Nominal test chemical concentration used for the study were 0, 5, 10, 20, 40 and 80 mg/l, respectively.
Details on test conditions:
TEST SYSTEM
- Test vessel: 50 ml glass vessel
- fill volume: 25 ml
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Natural water (surface or ground water), reconstituted water or dechlorinated tap water are acceptable as culturing and dilution water if D. magna survives in it for the duration of the culturing, acclimation and testing without showing signs of stress. Waters in the range pH 6 to pH 9, with hardness between 140 mg/l and 275 mg/l (as CaCO3) are recommended.
As an example, the preparation of dilution water meeting the requirements is described below.
Dissolve known quantities of reagents in water. The dilution water prepared shall have a pH of 7.8 ± 0.5, a hardness of (225 ± 50) mg/l (expressed as CaCO3), a molar Ca + Mg ratio close to 4 + 1 and a dissolved oxygen concentration above 7 mg/l.

Prepare the solutions specified below:
- Calcium chloride solution: Dissolve 117.6 g of calcium chloride dihydrate (CaCl2.2H2O) in water (4.2) and make up to 1 l with water (4.2).
- Magnesium sulfate solution: Dissolve 49.3 g of magnesium sulfate heptahydrate (MgSO4.7H2O) in water (4.2) and make up to 1 l with water (4.2).
- Sodium bicarbonate solution: Dissolve 25.9 g of sodium bicarbonate (NaHCO3) in water (4.2) and make up to 1 l with water (4.2).
- Potassium chloride solution: Dissolve 2.3 g of potassium chloride (KCI) in water (4.2) and make up to 1 l with water (4.2).

Mixing
Mix 2.5 ml of each of the four solutions and make up to 1 l with water.
The dilution water shall be aerated until the dissolved oxygen concentration has reached saturation and the pH has stabilized. If necessary, adjust the pH to 7.8 ± 0.5 by adding sodium hydroxide (NaOH) solution or hydrochloric acid (HCI). The dilution water prepared in this way shall not be further aerated before use.

- Sodium hydroxide solution, e.g. [NaOH] : 1 mol/l.
- Hydrochloric acid, e.g. [HCl] : 1 mol/l.

Reference substance:
Dissolve 600 mg of potassium dichromate (K2Cr2O7) in water and make up to 1 l with water (4.2).

OTHER TEST CONDITIONS
- Adjustment of pH: no adjustment done
- Photoperiod: No - Darkness
- Light intensity:

CALCULATION:
EC50 was calculated using non linear regression by the software Prism 4.0
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (K2Cr2O7)
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
30.1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95 % C. I. = 23.9 to 37.9 mg/l
Results with reference substance (positive control):
- Results with reference substance valid
- EC50: 0.73 mg/L (24 hours)
Reported statistics and error estimates:
EC50 was calculated using non linear regression by the software Prism 4.0.

In control solution containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test.

Result of the definitive test:

Sample no.

Sample Information

Conc. (mg/l)

I%

48 hr EC50

mg/l

95% C. I. (mg/l)

Control

Reconstituted water

0

0

 

 

14676

Test chemical

5

10

20

40

80

5

10

25

65

100

30.1

23.9 to 37.9

Validity criteria fulfilled:
yes
Conclusions:
Based on the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr EC50 value was determined to be 30.1 mg/l (95 % C. I. - 23.9 to 37.9 mg/l).
Executive summary:

An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on aquatic invertebrates. The test was performed in accordance to OECD guideline No. 202 “Daphnia sp.,Acute Immobilization Test”. Daphnia magna was used as a test organism for the study. The stock solution 100 mg/l was prepared by dissolving test chemical in reconstituted water. The test solutions of required concentrations were prepared by mixing the stock solution of the test sample in reconstituted water. Test chemical concentrations were not verified analytically. Nominal test chemical concentrations used for the study were 0, 10, 20, 40 and 80 mg/l, respectively. Study was performed using total 5 organisms per vessel/replicates in a static system. Daphnids were exposed to test chemical in 50 ml glass vessel in a volume of 25 ml of liquid solution containing both the chemical and media. Control solution vessel containing reconstituted water without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 20±1°C. With the test substance one positive control Potassium dichromate (K2Cr2O7) was run simultaneously. The 24 hr EC50 value of the reference substance was determined to be 0.73 mg/l. EC50 was calculated using non linear regression by the software Prism 4. In the control vessel containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test and the dissolved oxygen concentration at the end of the test was evaluated to be ≥ 3 mg/l (i.e, reported as > 7 mg/l) both in the control and test vessels. Thus, the validity criterion of the test has been fulfilled. On the basis of the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr median effect concentration (EC50) value was determined to be 30.1 mg/l (95 % C. I. - 23.9 to 37.9 mg/l). Thus, test chemical is considered as toxic to aquatic invertebrates at environmental related concentrations and hence, considered to be classified in 'aquatic chronic category 3' as per the CLP classification criteria.

Description of key information

An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on aquatic invertebrates (Experimental study report, 2018). The test was performed in accordance to OECD guideline No. 202 “Daphnia sp.,Acute Immobilization Test”. Daphnia magna was used as a test organism for the study. The stock solution 100 mg/l was prepared by dissolving test chemical in reconstituted water. The test solutions of required concentrations were prepared by mixing the stock solution of the test sample in reconstituted water. Test chemical concentrations were not verified analytically. Nominal test chemical concentrations used for the study were 0, 10, 20, 40 and 80 mg/l, respectively. Study was performed using total 5 organisms per vessel/replicates in a static system. Daphnids were exposed to test chemical in 50 ml glass vessel in a volume of 25 ml of liquid solution containing both the chemical and media. Control solution vessel containing reconstituted water without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 20±1°C. With the test substance one positive control Potassium dichromate (K2Cr2O7) was run simultaneously. The 24 hr EC50 value of the reference substance was determined to be 0.73 mg/l. EC50 was calculated using non linear regression by the software Prism 4. In the control vessel containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test and the dissolved oxygen concentration at the end of the test was evaluated to be ≥ 3 mg/l (i.e, reported as > 7 mg/l) both in the control and test vessels. Thus, the validity criterion of the test has been fulfilled. On the basis of the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr median effect concentration (EC50) value was determined to be 30.1 mg/l (95 % C. I. - 23.9 to 37.9 mg/l). Thus, test chemical is considered as toxic to aquatic invertebrates at environmental related concentrations and hence, considered to be classified in 'aquatic chronic category 3' as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
30.1 mg/L

Additional information

Experimental study of the test chemical and various supporting weight of evidence studies for its functionally similar read across chemical were reviewed for the short term toxicity to aquatic invertebrate end point which are summarized as below:

 

In an experimental study from study report (2018), an acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on aquatic invertebrates. The test was performed in accordance to OECD guideline No. 202 “Daphnia sp.,Acute Immobilization Test”. Daphnia magna was used as a test organism for the study. The stock solution 100 mg/l was prepared by dissolving test chemical in reconstituted water. The test solutions of required concentrations were prepared by mixing the stock solution of the test sample in reconstituted water. Test chemical concentrations were not verified analytically. Nominal test chemical concentrations used for the study were 0, 10, 20, 40 and 80 mg/l, respectively. Study was performed using total 5 organisms per vessel/replicates in a static system. Daphnids were exposed to test chemical in 50 ml glass vessel in a volume of 25 ml of liquid solution containing both the chemical and media. Control solution vessel containing reconstituted water without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 20±1°C. With the test substance one positive control Potassium dichromate (K2Cr2O7) was run simultaneously. The 24 hr EC50 value of the reference substance was determined to be 0.73 mg/l. EC50 was calculated using non-linear regression by the software Prism 4. In the control vessel containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test and the dissolved oxygen concentration at the end of the test was evaluated to be ≥ 3 mg/l (i.e, reported as > 7 mg/l) both in the control and test vessels. Thus, the validity criterion of the test has been fulfilled. On the basis of the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr median effect concentration (EC50) value was determined to be 30.1 mg/l (95 % C. I. - 23.9 to 37.9 mg/l).

 

In a supporting weight of evidence study, short term toxicity to aq. invertebrate study was conducted for 48 hrs for assessing the effect of test chemical (authoritative database, 2019 and secondary source, 2019). This test was performed using Daphnia magna (water flea) of < 24 hr old under static conditions. No analytical monitoring was performed of exposed test chemical concentrations. On the basis of effect on mobility of the test organism Daphnia magna, the 48 hr EC50 was determined to be 17 mg/l (nominal concentration).

 

For the test chemical from peer reviewed journal (A.M. Api et. al., 2019), short term toxicity to aquatic invertebrate study was conducted for 48 hrs for assessing the effect of test chemical. The study was performed following the American Society of Testing and Materials (ASTM) method in a static system. Daphnia magna was used as a test organism. On the basis of effect on mobility of the test organism Daphnia magna, the 48 hr EC50 was determined to be 30 mg/l (nominal concentration).

 

On the basis of the above results, it can be concluded that the test chemical can be considered as toxic to aquatic invertebrates at environmental related concentrations and considered to be classified in 'aquatic chronic category 3' as per the CLP classification criteria.

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