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Ecotoxicological information

Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well documented publication
Justification for type of information:
Read across is based on a structurally similar material. An analogue justification is attached in section 13 of dataset.
Reason / purpose for cross-reference:
read-across source
Principles of method if other than guideline:
The static water acute toxicity tests followed the methods described in the EPA publication, "Methods for Acute Toxicity Tests with Fish, Macroinvertebrates, and Amphibians" (COMMITTEE ON METHODS FOR TOXICITY WITH AQUATIC ORGANISMS, 1975).
GLP compliance:
no
Analytical monitoring:
not specified
Vehicle:
not specified
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
The test fish were obtained from the National Fish Hatchery at Hebron, Ohio, USA.
The fish had an average weight of 0.74 g (range 0.62-1.28 g) and an average total length of 34 mm (range 28-38 mm).
The fish were kept in a 16 -h light/8-h dark cycle. Feed was stopped three days prior to the bioassay to empty the digestive tract. Fish were placed into the aquaria 24 h before adding the test material.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
Three different water types were used:
very soft water: 10-13 mg/L CaCO3, reconstituted from destilled water
medium hard water: 103 mg/L as CaCO3, dilution water from Lake Huron
very hard water: 280-320 mg/L CaCO3, reconstituted from destilled water
Test temperature:
22 +/- 1°C
pH:
Three different water types were used:
very soft water reconstituted from destilled water, start: pH 6.4 - 6.8, end: unknown but low
medium hard water dilution water from Lake Huron: start pH 7.6, end pH 4
very hard water reconstituted from destilled water: start pH 8.0 - 8.4, end pH 4
Dissolved oxygen:
start of test: 8.2 mg/L,
end of test: 4.2 mg/L;
Salinity:
no data available
Nominal and measured concentrations:
no data available
Details on test conditions:
Water used came from Whitestone Point on the west side of Saginaw Bay, Lake Huron.
The water was sent through an activated charcoal bed to remove any free chlorine.
This water exhibits chemical characteristics in the following ranges:
total alkalinity: 85 mg/L as CaC03
specific conductivity: 1770 umhos/cm
resistivity: ca. 570 ohms/cm
This water passes the general requirements for dilution water as stated in COMMITTEE ON METHODS FOR TOXICITY WITH AQUATIC ORGANISMS (1975). In addition, VERSENE acid, VERSENE 100 were also tested for effect in very soft and hard waters. These waters were reconstituted distilled waters.

None of the products were neutralized prior to testing.

Bioassays were conducted by placing eight liters water in each vessel, a round glass aquarium measuring 22 cm deep with a 24.5 cm diameter, adding the fish, then aerating with plant air. If no deaths occurred in 24 h, the aerators were removed, and the test solution added with two liters of water for mixing, making a total of ten liters. Any vessel containing dead fish was cleaned, reset, and observed for another 24 h prior to compound addition. Ten fish were exposed to each concentration of the material. Between five and ten concentrations were set per product. Observations were made and recorded daily.
Reference substance (positive control):
not specified
Duration:
96 h
Dose descriptor:
other: NAEL
Effect conc.:
24 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: very soft water
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
41 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 34 - 62 mg/L, very soft water
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
75 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: very soft water
Duration:
96 h
Dose descriptor:
other: NAEL
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: medium hard water
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
159 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 136-204 mg/L, medium hard water
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
240 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: medium hard water
Duration:
96 h
Dose descriptor:
other: NAEL
Effect conc.:
420 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: very hard water
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
532 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 473-598 mg/L, very hard water
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
750 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: very hard water
Details on results:
Instead of the NOEC, in this study the NAEL (= no (observable) adverse effect level) is quoted.
Results with reference substance (positive control):
no data
Sublethal observations / clinical signs:


Two products illustrate the effect of pH in medium hard water: VERSENE acid and VERSENE powder. The acid kills
100 % of the bluegill at 240 mg/l in water at a pH of 3.7.
Such acidity alone is lethal to bluegill (COMMITTEE ON
WATER QUALITY CRITERIA, 1972). If the acid is made into
the sodium salt, VERSENE powder, a 100 % mortality occurs
at 521 mg/l. The resultant pH of 8.9 is not itself toxic
to bluegill. A 50 % aqueous solution of this sodium salt,
VERSENE 100, causes 100 % mortality at 1120 mg/l at a pH
of 9.6. In hard water, total kill with the VERSENE acid
occurs at 750 mg/l and pH 3.5 (again, a pH related kill),
whereas VERSENE 100 killed all bluegill at 2400 mg/l and
pH 9.8. A pH level of 9.5-10.0 may cause some mortality;
values >10.0 are not tolerated by bluegill (COMMITTEE ON
WATER QUALITY CRITERIA, 1972).
Validity criteria fulfilled:
not specified
Conclusions:
The test results of H4EDTA show that there is a relation between water hardness and ecotoxicity. Tests performed in very soft (10-13 mg/l CaCO3) and medium hard water (103 mg/I CaCO3) showed a higher toxicity than those performed in very hard water. As the hardness increased, more of the chelator was converted to the Ca-complex, thus more chelating agent was required to exert a toxic effect-less uncomplexed EDTA is available. This was proofed by a reported LC50 of 2340 mg/L for Versene Ca (where EDTA is already complexed with Ca) in the same publication. Thus, it should be noted, that under test conditions (buffer capacity) pH 4 alone leads to toxicity in bluegill.
Endpoint:
short-term toxicity to fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well documented publication
Justification for type of information:
Read across is based on a structurally similar material. An analogue justification is attached in section 13 of dataset.
Reason / purpose for cross-reference:
read-across source
Principles of method if other than guideline:
The static water acute toxicity tests followed the methods described in the EPA publication, "Methods for Acute Toxicity Tests with Fish, Macroinvertebrates, and Amphibians" (COMMITTEE ON METHODS FOR TOXICITY WITH AQUATIC ORGANISMS, 1975).
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
no data available
Analytical monitoring:
not specified
Details on sampling:
no data available
Vehicle:
not specified
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
The test fish were obtained from the National Fish Hatchery at Hebron, Ohio, USA.
The fish had an average weight of 0.74 g (range 0.62-1.28 g) and an average total length of 34 mm (range 28-38 mm).
The fish were kept in a 16 -h light/8-h dark cycle. Feed was stopped three days prior to the bioassay to empty the digestive tract. Fish were placed into the aquaria 24 h before adding the test material.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
no data available
Hardness:
Three different water types were used:
very soft water: 10-13 mg/L CaCO3, reconstituted from destilled water
medium hard water: 103 mg/L as CaCO3, dilution water from Lake Huron
very hard water: 280-320 mg/L CaCO3, reconstituted from destilled water
Test temperature:
22 +/- 1°C
pH:
Three different water types were used:
very soft water reconstituted from destilled water, start: pH 6.4 - 6.8, end: 7.5
medium hard water dilution water from Lake Huron: start pH 7.6, end pH 9.5
very hard water reconstituted from destilled water: start pH 8.0 - 8.4, end pH 9.5
Dissolved oxygen:
start of test: 8.2 mg/L,
end of test: 4.2 mg/L;
Salinity:
no data
Nominal and measured concentrations:
no data
Details on test conditions:
TEST WATER
Water used came from Whitestone Point on the west side of Saginaw Bay, Lake Huron.
The water was sent through an activated charcoal bed to remove any free chlorine.
This water exhibits chemical characteristics in the following ranges:
total alkalinity: 85 mg/L as CaC03;
specific conductivity: 1770 µmhos/cm; and resistivity: ca. 570 ohms/cm (AMERICAN PUBLIC HEALTH ASSOCIATION 1975).
This water passes the general requirements for dilution water as stated in COMMITTEE ON METHODS FOR TOXICITY WITH AQUATIC ORGANISMS (1975).
None of the products were neutralized prior to testing.
Bioassays were conducted by placing eight liters water in each vessel, a round glass aquarium measuring 22 cm deep with a 24.5 cm diameter, adding the fish, then aerating with plant air. If no deaths occurred in 24 h, the aerators were removed, and the test solution added with two liters of water for mixing, making a total of ten liters. Any vessel containing dead fish was cleaned, reset, and observed for another 24 h prior to compound addition. Ten fish were exposed to each concentration of the material. Between five and ten concentrations were set per product. Observations were made and recorded daily.
Reference substance (positive control):
not specified
Duration:
96 h
Dose descriptor:
other: NAEL
Effect conc.:
88 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na4EDTA
Basis for effect:
mortality (fish)
Remarks on result:
other: very soft water
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
121 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na4EDTA
Basis for effect:
mortality (fish)
Remarks on result:
other: 113-130, very soft water
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
138 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na4EDTA
Basis for effect:
mortality (fish)
Remarks on result:
other: very soft water
Duration:
96 h
Dose descriptor:
other: NAEL
Effect conc.:
669 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na4EDTA
Basis for effect:
mortality (fish)
Remarks on result:
other: medium hard water
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
792 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na4EDTA
Basis for effect:
mortality (fish)
Remarks on result:
other: 754-831, medium hard water
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
861 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na4EDTA
Basis for effect:
mortality (fish)
Remarks on result:
other: medium hard water
Duration:
96 h
Dose descriptor:
other: NAEL
Effect conc.:
1 380 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na4EDTA
Basis for effect:
mortality (fish)
Remarks on result:
other: hard water
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
1 592 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na4EDTA
Basis for effect:
mortality (fish)
Remarks on result:
other: 1493-1678, hard water
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
1 846 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na4EDTA
Basis for effect:
mortality (fish)
Remarks on result:
other: hard water
Details on results:
Instead of the NOEC, in this study the NAEL (= no (observable) adverse effect level) is quoted.
Results with reference substance (positive control):
no data available
Validity criteria fulfilled:
not specified
Conclusions:
The test results of Na4EDTA show that there is a relation between water hardness and ecotoxicity. Tests performed in very soft water (10-13 mg/l CaCO3) showed a higher toxicity than those performed in a medium hard water (103 mg/I CaCO3) or very hard water (280-320 mg CaCO3). The great variation in mortality resulted in LC50 values in a range of 121 mg/L (soft water) to 1592 mg/L (very hard water). The LC50 of 121 mg/L derived in very soft water which reflects spring water from lime-free geological zones is not considered as environmentally relevant as EDTA is not released to waters of such low hardness.
Endpoint:
short-term toxicity to fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well documented publication
Justification for type of information:
Read across is based on a structurally similar material. An analogue justification is attached in section 13 of dataset.
Reason / purpose for cross-reference:
assessment report
Principles of method if other than guideline:
The static water acute toxicity tests followed the methods described in the EPA publication, "Methods for Acute Toxicity Tests with Fish, Macroinvertebrates, and Amphibians" (COMMITTEE ON METHODS FOR TOXICITY WITH AQUATIC ORGANISMS, 1975).
GLP compliance:
no
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
no data available
Analytical monitoring:
not specified
Details on sampling:
no data available
Vehicle:
not specified
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
The test fish were received from the National Fish Hatchery at Hebron, Ohio, USA.
The fish had an average weight of 0.74 g (range 0.62-1.28 g) and an average total length of 34 mm (range 28-38 mm).
The fish were kept in a 16 -h light/8-h dark cycle. PFeed was stopped three days prior to the bioassay to empty the digestive tract. Fish were placed into the aquaria 24 h before adding the test material.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
no data available
Hardness:
103 mg/L as CaCO3, dilution water from Lake Huron
Test temperature:
22 +/- 1°C
pH:
start pH 7.6, end pH 8.0
Dissolved oxygen:
start of test: 8.2 mg/L,
end of test: 4.2 mg/L;
Salinity:
no data
Nominal and measured concentrations:
no data
Details on test conditions:
TEST WATER
Water used in the fish laboratory came from Whitestone Point on the west side of Saginaw Bay, Lake Huron.
The water was sent through an activated charcoal bed to remove any free chlorine.
This water exhibits chemical characteristics in the following ranges:
total alkalinity: 85 mg/L as CaC03;
specific conductivity: 1770 µmhos/cm; and resistivity: ca. 570 ohms/cm (AMERICAN PUBLIC HEALTH ASSOCIATION 1975).
This water passes the general requirements for dilution water as stated in COMMITTEE ON METHODS FOR TOXICITY WITH AQUATIC ORGANISMS (1975).
None of the products were neutralized prior to testing.
Bioassays were conducted by placing eight liters water in each vessel, a round glass aquarium measuring 22 cm deep with a 24.5 cm diameter, adding the fish, then aerating with plant air. If no deaths occurred in 24 h, the aerators were removed, and the test solution added with two liters of water for mixing, making a total of ten liters. Any vessel containing dead fish was cleaned, reset, and observed for another 24 h prior to compound addition. Ten fish were exposed to each concentration of the material. Between five and ten concentrations were set per product. Observations were made and recorded daily.
Reference substance (positive control):
not specified
Duration:
96 h
Dose descriptor:
other: NAEL
Effect conc.:
240 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
705 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: 623-795
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
Instead of the NOEC, in this study the NAEL (= no (observable) adversed effect level) is quoted.
Validity criteria fulfilled:
not specified
Conclusions:
In an acute fish test with (NH4)4EDTA the LC50 of Lepomis macrochirus was determined to be 705 mg/L after 96 h.

Description of key information

With high probability the test substance is acutely not harmful to fish.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
1 000 mg/L

Additional information

The toxicity of EDTA on fish highly depends on water hardness, pH and metal speciation [EU Risk Assessment, 2004]. The toxicity of EDTA complexes to bluegill was determined in a key study performed by Batchelder et al. (1980), which takes into account the water hardness and pH. The revealed LC50 -values are in a range of 41 mg/L to 2070 mg/L.

Two consequences can be drawn. First, the tests performed with the acid form lead to very low pH values under test conditions. A pH of 4 alone leads to toxicity in bluegill. The low LC50 value in very soft water can be explained by a surplus of uncomplexed EDTA which was present in the test media. This is not expected in the environment; therefore the results derived from studie under conditions of soft water hardness and/or pH <4 are not relevant for the assessment.

The 96-hour LC50 value of (NH4)4EDTA was determined to be 705 mg/L. at pH 8. The toxicity is driven by ammonia. A

t pH 8 ca. 5.3 % of the total ammonia are present as free ammonia, resulting in a NH3 concentration of 2.3 mg/L. Generally a NH3 concentration above 1 mg/L can be considered as toxic alone to fish. Thus, the results of the experiment with (NH4)4 EDTA are also not relevant for the risk assessment.

 

Consequently the LC50 of EDTA is estimated to be higher than 1000 mg/L, due to the fact that this predicted value represents the lowest LC50 at environmentally relevant pH which has been performed in natural not synthetic water. The results indicate that for complexed and non complexed EDTA no need for classification as dangerous for the environment. Further on, the results show that the toxicity of complexes with the toxic metals Cu and Zn is in the same concentration range as the uncomplexed EDTA. Chelates with non-toxic metals (Mn, Ca, Mg) are much less toxic.

For justification for read-across please refer to IUCLID 5, Chapter 13.