potassium;2-[bis[2-[bis(carboxymethyl)amino]ethyl]amino]acetate;dicalcium

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE CATEGORY APPROACH

1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
See the documentation in support for the category approach.

2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL
See the documentation in support for the category approach.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

Non critical deviation

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: the control, the lowest, the highest concentrations and a concentration around the expected EC50 concentration: 0, 2, 20.7 and 100 mg/L
- Sampling method: All the solutions will be analysed taken directly from the starting solutions without daphnids.
- Sample storage conditions before analysis: All the solutions were kept in refrigerator or freeze in order to be analyzed if necessary or relevant.

Vehicle:

no

Details on test solutions:

According to the sponsor, the test item is soluble in water. The dilution water was the daphnids test medium, a weakly saline aqueous medium.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name:
- Strain/clone: clone number 5
- Justification for species other than prescribed by test guideline: not relevant
- Source: laboratory culture
- Age of parental stock (mean and range, SD): not specified
- Feeding during test: no
- Food type: not specified
- Amount: not specified
- Frequency: not specified

ACCLIMATION
no acclimation

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

19.8 – 20.0°C

pH:

7.6-7.9

Dissolved oxygen:

8.9-9.3 mg/L

Nominal and measured concentrations:

Nominal concentration (mg/L): 2.0, 4.3, 9.4, 20.7, 45.5, 100.0.
Measured concentrations: see table below

Details on test conditions:

TEST SYSTEM
- Test vessel: tube
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass tubes of 10 ml
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): not relevant
- Renewal rate of test solution (frequency/flow rate): no renewal
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): not relevant
- Biomass loading rate: not specified

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ultrapure water
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: not needed
- Photoperiod: total darkness
- Light intensity: not relevant

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
immobilization (24 and 48 hours)

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Test concentrations: 0, 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

64.74 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

18.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No sign of stress was observed on the mobile daphnids after 48 hours of exposure to the highest concentration.

Results with reference substance (positive control):

The last toxic response obtained on the reference item K2Cr2O7 (November 2, 2017) gives an EC50-24h of 1.38 mg/L (1.26 – 1.51), indicating that the daphnids sensitivity is correct (within the accepted interval of 0.6 – 2.1 mg/L fixed by the International Standard ISO 6341 December 2012 as mentioned in the OECD 202 Guideline) and in accordance with historical data obtained by the laboratory.

Reported statistics and error estimates:

See table.

Validity criteria for the test

	Criteria for OECD 202	Validity criteria conformity
Immobilised daphnids in the Control	<10%	Yes (0%)
Dissolved Oxygen at the end of the test	>3 mg/L	Yes (8.9 mg/L)

EC50 values for immobilization

Observation time	24h	48h
EC50in mg/l [Confidence interval]	64.74 mg/L [53.84 – 78.15]	18.05 mg/L [14.53 – 22.66]

Validity criteria fulfilled:

yes

Conclusions:

All the validity criteria were successful.

The results obtained for the test item “REACTION MIXTURE OF CuDTPA and CuHEEDTA” on the toxicological endpoint “immobilisation” give an EC50-24h = 64.74 mg/L [53.84 – 78.15] and an EC50-48h = 18.05 mg/L [14.53 22.66] based on nominal test item concentrations.

Executive summary:

This study was carried out in accordance with the study plan no.17/1108F/d dated November 6, 2017.

There was one deviation (cf. 9.3) to the study plan during this study.

There was no amendment during this study.

 

The aim of this study was to assess the effects of the test item “REACTION MIXTURE OF CuDTPA and CuHEEDTA”on the daphnidsDaphnia magna, according to the OECD 202 guideline, in static conditions.

 

The concentrations which immobilise 50% of the daphnids exposed to the test item after 24 and 48 hours, reported as EC₅₀-24h and EC₅₀-48h were evaluated, relative to the tested concentrations.

 

According to the sponsor, chemical analyses of “REACTION MIXTURE OF CuDTPA and CuHEEDTA” in the test solutions was performed by ICP/OES in order to check the test item stability throughout the study.

The test item « REACTION MIXTURE OF CuDTPA and CuHEEDTA» was considered stable under test conditions because the measured concentrationsof Copper (Cu) in samples containing the test itemdid not vary more than ± 20 % during the test.

 

The values are based on nominal test item “REACTION MIXTURE OF CuDTPA and CuHEEDTA” concentration.

 

Validity criteria conformity:

	Values for OECD 202	Validity criteria for the test
Immobilised daphnids in control	<10%	Yes (0%)
Dissolved Oxygen concentration	>3 mg/L	Yes (8.9 mg/L)

 

All the validity criteria were successful.

 

Daphnids immobilisation

Nominal test item concentrations in mg/L	Number of daphnids exposed at the start	Immobilised daphnids (%)
		at 24h	at 48h
Control	20	0 (0%)	0 (0%)
2.0	20	0 (0%)	0 (0%)
4.3	20	0 (0%)	0 (0%)
9.4	20	0 (0%)	3 (15%)
20.7	20	0 (0%)	10 (50%)
45.5	20	3 (15%)	20 (100%)
100.0	20	18 (90%)	20 (100%)

 

 

EC₅₀values at different observation times

Observation time	24h	48h
EC50in mg/l [Confidence interval]	64.74 mg/L [53.84 – 78.15]	18.05 mg/L [14.53 – 22.66]

The results obtained for the test item “REACTION MIXTURE OF CuDTPA and CuHEEDTA” on the toxicological endpoint “immobilisation” give an EC₅₀-24h = 64.74 mg/L [53.84 – 78.15] and an EC₅₀-48h = 18.05 mg/L [14.53 - 22.66]based on nominal test item concentrations.

 

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE CATEGORY APPROACH

1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
See the documentation in support for the category approach.

2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL
See the documentation in support for the category approach.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

Non critical deviation

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: the control, the lowest, the highest concentrations and a concentration around the expected EC50 concentration: 0, 10, 231 and 1118 mg/L
- Sampling method: All the solutions will be analysed taken directly from the starting solutions without daphnids.
- Sample storage conditions before analysis: All the solutions were kept in refrigerator or freeze in order to be analyzed if necessary or relevant.

Vehicle:

no

Details on test solutions:

According to the sponsor, the test item is soluble in water. The dilution water was the daphnids test medium, a weakly saline aqueous medium.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name:
- Strain/clone: clone number 5
- Justification for species other than prescribed by test guideline: not relevant
- Source: laboratory culture
- Age of parental stock (mean and range, SD): not specified
- Feeding during test: no
- Food type: not specified
- Amount: not specified
- Frequency: not specified

ACCLIMATION
no acclimation

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

19.8 – 20.0°C

pH:

6.5-7.9

Dissolved oxygen:

8.8-9.1 mg/L

Nominal and measured concentrations:

Nominal concentration (mg/L): 0, 10, 20, 48, 105, 231, 508, 1118
Measured concentrations: see table below

Details on test conditions:

TEST SYSTEM
- Test vessel: tube
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass tubes of 10 ml
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): not relevant
- Renewal rate of test solution (frequency/flow rate): no renewal
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): not relevant
- Biomass loading rate: not specified

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ultrapure water
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: not needed
- Photoperiod: total darkness
- Light intensity: not relevant

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
immobilization (24 and 48 hours)

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Test concentrations: 0, 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 1 118 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 1 118 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No sign of stress was observed on the mobile daphnids after 48 hours of exposure to the highest concentration.

Results with reference substance (positive control):

The last toxic response obtained on the reference item K2Cr2O7 (November 2, 2017) gives an EC50-24h of 1.38 mg/L (1.26 – 1.51), indicating that the daphnids sensitivity is correct (within the accepted interval of 0.6 – 2.1 mg/L fixed by the International Standard ISO 6341 December 2012 as mentioned in the OECD 202 Guideline) and in accordance with historical data obtained by the laboratory.

Reported statistics and error estimates:

See table.

Validity criteria for the test

	Criteria for OECD 202	Validity criteria conformity
Immobilised daphnids in the Control	<10%	Yes (0%)
Dissolved Oxygen at the end of the test	>3 mg/L	Yes (8.9 mg/L)

 

All the validity criteria were successful.

EC50 values for immobilization

Observation time	24h	48h
EC50in mg/l	>1118 mg/L	>1118 mg/L

 

Validity criteria fulfilled:

yes

Conclusions:

All the validity criteria were successful.

The results obtained for the test item “REACTION MIXTURE OF ZnEDTA, ZnDTPA AND ZnHEEDTA ” on the toxicological endpoint “immobilisation” give an EC50-24h and an EC50-48h higher than 1118 mg/l (highest tested concentration) based on nominal test item concentrations.

Executive summary:

This study was carried out in accordance with the study plan no.17/1107F/f dated October 24, 2017.

There was no deviation and no amendment to the study plan during this study.

 

The aim of this study was to assess the effects of the test item “REACTION MIXTURE OF ZnEDTA, ZnDTPA AND ZnHEEDTA” on the daphnids Daphnia magna, according to the OECD 202 guideline, in static conditions.

 

The concentrations which immobilise 50% of the daphnids exposed to the test item after 24 and 48 hours, reported as EC₅₀-24h and EC₅₀-48h were evaluated, relative to the tested concentrations.

 

According to the sponsor, chemical analyses of “REACTION MIXTURE OF ZnEDTA, ZnDTPA AND ZnHEEDTA” in the test solutions was performed by ICP/OES in order to check the test item stability throughout the study.

 

The test item « REACTION MIXTURE OF ZnEDTA, ZnDTPA AND ZnHEEDTA» was considered stable under test conditions because the measured concentrationsof Zinc (Zn) in samples containing the test itemdid not vary more than ± 20 % during the test.

 

The values are based on nominal test item “REACTION MIXTURE OF ZnEDTA, ZnDTPA AND ZnHEEDTA”concentration.

 

 

Validity criteria conformity:

	Values for OECD 202	Validity criteria for the test
Immobilised daphnids in control	<10%	Yes (0%)
Dissolved Oxygen concentration	>3 mg/L	Yes (8.9 mg/L)

 

All the validity criteria were successful.

 

Daphnids immobilisation

Nominal test item concentrations in mg/L	Number of daphnids exposed at the start	Immobilised daphnids (%)
		at 24h	at 48h
Control	20	0 (0%)	0 (0%)
10	20	0 (0%)	0 (0%)
22	20	0 (0%)	0 (0%)
48	20	0 (0%)	0 (0%)
105	20	0 (0%)	0 (0%)
231	20	0 (0%)	0 (0%)
508	20	0 (0%)	0 (0%)
1118	20	0 (0%)	0 (0%)

 

 

EC₅₀values at different observation times

Observation time	24h	48h
EC50in mg/l	>1118 mg/L	>1118 mg/L

 

The results obtained for the test item “REACTION MIXTURE OF ZnEDTA, ZnDTPA AND ZnHEEDTA” on the toxicological endpoint “immobilisation” give an EC₅₀-24h and an EC₅₀-48h higher than 1118 mg/L (highest tested concentration) based on nominal test item concentrations.

Description of key information

Information on the short-term toxicity to aquatic invertebrates is available for two category members: metal chelates with copper and zinc. The result is in line with the expectations i.e. that the aquatic toxicity is the result of the free (metal) ion that is complexed and that this ion is much less bioavailable and thus less toxic to aquatic life when complexed with chelating agents. As expected, the order of aquatic toxicity of the metal chelates follows the order of toxicity of the metal ions: copper chelate is more toxic than zinc chelate. Therefore, it is reasonable to assume that the aquatic toxicity of KCa2DTPA will be lower than that of zinc chelate.

The EC50 for Daphnia magna (OECD 202) was determined to exceed 1118 mg/L for the zinc chelate.

Considering that the aquatic toxicity is determined by the metal ion and that K and Ca are orders of magnitude less toxic than Zn, the substance is not considered toxic to aquatic invertebrates.