Iron(II) fumarate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed using standard 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 in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test 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

ACCLIMATION - No data available

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:

pH at higher test concentration 100 mg/l: 7.1 changes to 6.2
Control: 8 did not change during test

Dissolved oxygen:

higher than 8.1 mg/L at the end of test

Nominal and measured concentrations:

Nominal concentrations: 0, 10, 18, 32, 58 and 100 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.:

56.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI was 42.2 - 75.2 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

Validity criteria fulfilled:

yes

Conclusions:

After the exposure period of 48 hours of test chemical with fresh water aquatic invertebrates Daphnia magna, 50 % immobility were observed. The median effective concentration i.e. EC50 value of the test chemical on Daphnia magna was determined to be 56.3 mg/L.

Executive summary:

Principle of this study was to determine the effect of test chemical on aquatic invertebrates. Test conducted in accordance with the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna used as a test organism. The stock solution 100 mg/l was prepared in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. Nominal concentrations 0, 10, 18, 32, 58 and 100 mg/l, respectively were used. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. Temperature, pH and dissolved oxygen were maintained at 20±1°C, 6.2 to 7.1, higher than 8.1 mg/L at the end of test. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. In each glass beaker > 24 hrs old 5 Daphnia magna were added. Test performed in the four replicates. Observations for immobilized Daphnids were recorded in the interval of 24 hours (0, 24 and 48 hours of exposure). After the exposure period of 48 hours of test chemical with fresh water aquatic invertebrates Daphnia magna, 50 % immobility were observed. The median effective concentration i.e. EC50 value of the test chemical on Daphnia magna was determined to be 56.3 mg/L. Thus, based on the EC50 value, test chemical consider to be toxic and classified in aquatic chronic category 3 as per the CLP classification criteria. But as the test chemical is readily biodegradable in water, thus consider to be nontoxic and not classified as per the CLP classification criteria.

Description of key information

Principle of this study was to determine the effect of test chemical on aquatic invertebrates. Test conducted in accordance with the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna used as a test organism. The stock solution 100 mg/l was prepared in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. Nominal concentrations 0, 10, 18, 32, 58 and 100 mg/l, respectively were used. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. Temperature, pH and dissolved oxygen were maintained at 20±1°C, 6.2 to 7.1, higher than 8.1 mg/L at the end of test. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. In each glass beaker > 24 hrs old 5 Daphnia magna were added. Test performed in the four replicates. Observations for immobilized Daphnids were recorded in the interval of 24 hours (0, 24 and 48 hours of exposure). After the exposure period of 48 hours of test chemical with fresh water aquatic invertebrates Daphnia magna, 50 % immobility were observed. The median effective concentration i.e. EC50 value of the test chemical on Daphnia magna was determined to be 56.3 mg/L. Thus, based on the EC50 value, test chemical consider to be toxic and classified in aquatic chronic category 3 as per the CLP classification criteria. But as the test chemical is readily biodegradable in water, thus consider to be nontoxic and not classified as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

56.3 mg/L

Additional information

Summarized result from the various experimental sources for the determination of effect of test chemical and structurally and functionally similar read across chemicals on the mobility of aquatic invertebrates are as mentioned below:

 

Principle of this study was to determine the effect of test chemical on aquatic invertebrates. Test conducted in accordance with the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna used as a test organism. The stock solution 100 mg/l was prepared in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. Nominal concentrations 0, 10, 18, 32, 58 and 100 mg/l, respectively were used. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. Temperature, pH and dissolved oxygen were maintained at 20±1°C, 6.2 to 7.1, higher than 8.1 mg/L at the end of test. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. In each glass beaker > 24 hrs old 5 Daphnia magna were added. Test performed in the four replicates. Observations for immobilized Daphnids were recorded in the interval of 24 hours (0, 24 and 48 hours of exposure). After the exposure period of 48 hours of test chemical with freshwater aquatic invertebrates Daphnia magna, 50 % immobility were observed. The median effective concentration i.e. EC50 value of the test chemical on Daphnia magna was determined to be 56.3 mg/L. Thus, based on the EC50 value, test chemical considered to be toxic and classified in aquatic chronic category 3 as per the CLP classification criteria. But as the test chemical is readily biodegradable in water, thus consider to be nontoxic and not classified as per the CLP classification criteria.

 

Above experimental study further supported by the second study from experimental source. Principle of this study was to determine the effect of test chemical on aquatic invertebrates. Test conducted in accordance with the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna used as a test organism. The stock solution 120 mg/l was prepared in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. Nominal concentrations 0, 14, 23, 39, 70 and 120 mg/l, respectively were used. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. Temperature, pH and dissolved oxygen were maintained at 20±1°C, 7.1, higher than 8.7 mg/L. After the exposure of test chemical with Daphnia magna, effect concentration EC50 value was calculated using nonlinear regression by the software Prism 4.0. In each glass beaker > 24 hrs old 5 Daphnia magna were added. Test performed in the four replicates. Observations for immobilized Daphnids were recorded in the interval of 24 hours (0, 24 and 48 hours of exposure). After the exposure period of 48 hours of test chemical with aquatic invertebrates Daphnia magna, 50 % immobility were observed. The median effective concentration i.e. EC50 value of the test chemical on Daphnia magna was determined to be 67.9 mg/L with 95% CI of 56.6 to 81.3 mg/l. Thus, based on the EC50 value, test chemical considers to be toxic and classified in aquatic chronic category 3 as per the CLP classification criteria. But as the test chemical is readily biodegradable in water, thus consider to be nontoxic and not classified as per the CLP classification criteria.

 

Similarly, an acute study was performed to determine the effect of test chemical on the aquatic invertebrates. Test conducted according to the “Methods for Acute Toxicity Tests with Fish, macro invertebrates and Amphibians.” Daphnia magna was used as a test organism. Static system was used and chemical exposed for 48 hours with Daphnids. 5 daphnia were added in each vessel and test conducted in duplicates. Hardness, temperature, and pH were maintained at 154. 5mg/l CaCO3, 22 °C and 7.7. Immobility of Daphnia were used as a key parameter for the effect determination. Based on the immobility of Daphnia magna due to the test chemical exposure for 48 hours, the EC50 value was determined to be 212 mg/l. Thus, on the basis of EC50 value, test chemical was considered to be non-toxic to aquatic invertebrates and can be considered to be not classified as per the CLP regulations.

 

Hence, based on above all studies and effects observation, test chemical considers to be nontoxic and not classified as per the CLP classification criteria.