Trisodium 2-(carboxylatomethyl(2-hydroxyethyl)amino)ethyliminodi(acetate)

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Reasonably documented study although conducted prior to guideline publication; no analytical monitoring of test substance

Principles of method if other than guideline:

Fathead minnows were exposed for 96 hours in a static system to the test substance (assumed 100% purity). Dissolved oxygen in test vessels was not recorded, nor was the performance of controls.
There were no measured concentrations of test substance; results werebased on nominal concentrations. A pH of 4.0 was measured in the test vessels containing the substance at 560 mg/L. The acidic pH is believed primarily responsible for the kill as fish do not normally tolerate a pH outside of the range of 5-9.5.

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

Not reported

Vehicle:

no

Details on test solutions:

Not reported

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: fathead minnow
- All other template details: Not reported

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported

Hardness:

Not reported

Test temperature:

12 degrees C

pH:

The only reported pH was 4.0 measured in the 560 mg/L water during testing.

Dissolved oxygen:

Not reported

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal concentrations (mg/L): 180, 320, 560

Details on test conditions:

Not reported

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

372 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI = 299-466 mg/L

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

180 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

other: partial kill

Effect conc.:

320 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

other: 100% kill

Effect conc.:

560 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

271 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI = 149-327 mg/L

Duration:

96 h

Dose descriptor:

other: LC90

Effect conc.:

509 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI = 421-952 mg/L

Details on results:

The acidic pH which was measured in water solutions of this material is believed to be primarily responsible for the kill. A pH of 4.0 was measured in the test exposure aquarium containing 560 mg/L of the test compound in Lake Huron water. Fish normally cannot tolerate a pH outside the range 5-9.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

The LC values were calculated using a computer program of Finney's method of probit analysis, the slope was 9.5 (95% CI = 3.42-15.27).

Sublethal observations / clinical signs:

Not applicable

Validity criteria fulfilled:

not applicable

Remarks:

no guideline available at the time of this study

Conclusions:

The 96 hour LC50 for the fathead minnow was 372 mg/L Versenol acid chelating agent.

Executive summary:

Fathead minnows were exposed to the test substance under static conditions at 12 degrees C for 96 hours and the NOEC, LC10, LC50, LC90, as well as partial kill and 100% kill were assessed. LC values were calculated using Finney's method of probit analysis. The LC50 was 372 mg/L, indicating relatively low toxicity to fish. There was limited test condition data reported (no dissolved oxygen levels reported for example). Analytical confirmation of the test substance was not performed.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 to 29, 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

modified to marine conditions

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Details on sampling:

Not applicable

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: a nominal 1000 mg/L solution was prepared in dilution water, shaken vigorously and allowed to stand for one hour.
- Evidence of undissolved material: None - a clear colorless solution was obtained

Test organisms (species):

other: Scophthalmus maximus

Details on test organisms:

TEST ORGANISM
- Common name: Turbot
- Source: Mannin Sea Farms Limited, Derbyhaven, Castletown, Isle of Man
- Feeding during test: Not reported

ACCLIMATION
- Acclimation period: 8 to 25 October 1999
- Acclimation conditions: temperature 13.5 to 17 degrees C, dissolved oxygen >86% ASV
- Type and amount of food: Not reported

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported

Hardness:

Not reported

Test temperature:

12.5 - 15.0 degrees C

pH:

8.2-8.8

Dissolved oxygen:

92-98 % saturation

Salinity:

34 - 36 ppt

Nominal and measured concentrations:

Nominal (mg/L): 0, 560, 1000, 1800
(Note that higher concentrations were not tested due to insufficient sample available)

Details on test conditions:

TEST SYSTEM
- Test vessel: plastic aquaria
- Material, size, headspace, fill volume: 10 liters of test solution
- Renewal rate of test solution (frequency/flow rate): every 48 hours
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: standardized artificial seawater -Tropic Marin, formulated by Tropical Marine Centre Limited
- Salinity: 34 to 36 g/L sodium chloride
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: not reported
- Photoperiod: 16 hours light to 8 hours dark
- Light intensity: not reported

EFFECT PARAMETERS MEASURED : observations of mortality made every 24 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: about 2 x
- Range finding study
- Results used to determine the conditions for the definitive study: 48 hour LC50 > 1000 mg/L

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 800 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 738 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

41% purity

Basis for effect:

mortality (fish)

Details on results:

- Observations on body length and weight: mean length = 43.8 mm, mean weight = 2.15 g
- Mortality of test animals: 0 dead
- Mortality of control: 0 dead
- All other template details: Not reported

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

LC50 values were estimated and 95% confidence limits calculated using ToxCalc version 05 "Comprehensive Toxicity Data Analysis and Database Software"

Sublethal observations / clinical signs:

Not applicable

Validity criteria fulfilled:

yes

Conclusions:

The LC50 values at all time periods from 24 to 96 hours were estimated to be > 1,800 mg/L based on administered substance or >738 mg/L when corrected for the level of purity.

Executive summary:

The LC50 values at all time periods from 24 to 96 hours were estimated to be > 1,800 mg/L based on administered substance or >738 mg/L when corrected for the level of purity.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

July 23 - 27 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

other: DIN 38 412 "Testverfahren mit Wasserorganismen (Gruppe L).

Deviations:

not specified

Principles of method if other than guideline:

Ten golden orfe fish (Leucisus idus L., golden variety) are exposed to the test material for 96 hours dissolved in a static volume of water in a 10 L aquarium and observed for mortality. Fish were adapted to the test water for 3 days prior to the study, and food was withdrawn 1 day before and during exposure.

GLP compliance:

not specified

Analytical monitoring:

no

Details on sampling:

Not reported

Vehicle:

not specified

Details on test solutions:

The product was added to the test water without any pretreatment.

Test organisms (species):

Leuciscus idus

Details on test organisms:

TEST ORGANISM
- Common name: Golden orfe
- Strain: not reported
- Source: Fischzucht Paul Eggers, D-2354 Hohenwestedt, Frg
- Age at study initiation (mean and range, SD): not reported
- Length at study initiation: 9.8 cm (range 8.5 - 11.4)
- Weight at study initiation: 9.1 g (range 5.8 - 17.0)
- Method of breeding: not reported
- Feeding during test: not reported

ACCLIMATION
- Acclimation period: about 6 months, general; 3 days to test conditions
- Acclimation conditions: same as test for three days
- Type and amount of food: growing feed F/B 50, SSNIFF Spezialdialten GMBH, D-4770 SOEST, FRG
- Feeding frequency: ad libitum
- Health during acclimation (any mortality observed): mortality during last two weeks of housing, about 0.2%, mortality during adaptation period - 0%

QUARANTINE: Not applicable

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported

Hardness:

2.5 mmol/L

Test temperature:

20-21 degrees C

pH:

about 8.0

Dissolved oxygen:

Not reported

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal (mg/L): 0, 1000, 2150, 4640, 10000

Details on test conditions:

TEST SYSTEM
- Test vessel: all glass aquarium, 30 cm x 22 cm x 24 cm filled with 10 L of water
- Aeration: continuous aeration with oil-free water
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): not reported
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): not reported
- Biomass loading rate: 9.1 g fish / L test water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted freshwater, preparation from fully demineralized tap water
- Alkalinity: 63 mg/L NaHCO3
- KCl: 5.5 mg/L
- Ca/mg ratio: 4:1
- Na/K ratio: 10:1
- Conductivity: max 10 micro MHO
- All other template details: Not reported

OTHER TEST CONDITIONS
- Adjustment of pH: pH was adjusted at the 10,000 mg/L concentration with an unspecified compound
- Photoperiod: 16 hrs light : 8 hrs dark
- Light intensity: not reported

EFFECT PARAMETERS MEASURED: mortality at 1 hr, 4 hr, 24 hr, 48 hr, 72 hr, 96 hr

TEST CONCENTRATIONS
- Spacing factor for test concentrations: about 2.2
- Range finding study
- Test concentrations: 1,000 and 10,000 mg/L
- Results used to determine the conditions for the definitive study: LC 50 between 1,000 and 10,000 mg/L

Reference substance (positive control):

yes

Remarks:

chloroacetamide

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 2 150 - < 4 640 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 1% significance level

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 862 - < 1 861 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

at 40.1%

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

2 150 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

862 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

at 40.1%

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

4 640 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 100% mortality at this concentration

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

1 861 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

act. ingr.

Remarks:

at 40.1%

Basis for effect:

mortality (fish)

Details on results:

- See Table 1 for Measurements of pH, Oxygen content, and temperature measured during the study
- See Table 2 for summary of mortality observed during the study
- Behavioural abnormalities: no symptoms detectable at any observation period at all doses
- Mortality of control: 0/10
- All other template details: not reported

Results with reference substance (positive control):

- LC50 after 48 hr: about 31 mg/L, determined 7 May 1990
- This lethal concentration corresponds to the normal sensitivity

Reported statistics and error estimates:

LC50 calculated using probit analysis from Finney 1971, certain aspects of this method have been modified - the exact procedure can be seen from the standard operating procedures of the laboratory.

Sublethal observations / clinical signs:

Table 1. Measurements of pH, Oxygen Content, and Temperature during the study.

Nominal Conc. (mg/L)	1 Hr	4 Hr	24 Hr	48 Hr	72 Hr	96 Hr
	pH
0	7.4		7.7	7.6	7.7	7.7
1000	7.4		7.6	7.6	7.7	7.7
2150	7.6		7.5	7.7	7.8	7.7
4640	10.8
10000	11.3
10000 **	7.3	** after pH adjustment with unspecified compound
	Oxygen Content (mg/L)
0	6.5		7.5	7.4	7.8	8.1
1000	6.5		7.4	7.4	7.5	8.0
2150	6.6		7.1	7.5	7.4	7.3
4640	8.5
10000	8.9
	Temperature (degrees C)
0	21		20	21	21	21
1000	21		21	21	21	21
2150	21		21	21	21	21
4640	21
10000	21

Table 2. Observed Fish Toxicity

Nominal Concentration (mg/L)	Number of fish at start	1 Hr	4 Hr	24 Hr	48 Hr	72 Hr	96 Hr
0	10	0	0	0	0	0	0
1000	10	0	0	0	0	0	0
2150	10	0	0	0	0	0	0
4640	10	10
10000	10	10

Conclusions:

The 96hr LC50 of Trilon D Fluessig to the golden Orfe (Leuciscus idus L., golden variety) was between 2150 and 4640 mg/L. No fish died at 2150 mg/L, and all 10 fish had died within 1 hr at 4640 mg/L and above.

Executive summary:

Ten Golden Orfe fish (Leuciscus idus L., golden variety) were exposed to Trilon D Fluessig in static aquaria at 20 -21 degrees C for up to 96 hours. The observed LC50 was between 862 and 1861 mg/L. No fish died at 862 mg/L, and all 10 fish had died within 1 hr at 1861 mg/L and 4010 mg/L.

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

96 hours

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Reasonably documented study although conducted prior to guideline publication; no analytical monitoring of test substance

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Bluegill fish were exposed for 96 hours in a static system to the test compound. Dissolved oxygen in test vessels was not recorded, nor were the
performance of controls. There were no measured concentration of test substance; results based on nominal concentrations.

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

Test substance was very soluble in water, relatively stable, and not expected to bioconcentrate. The amount of Versenol 120 was not reduced after 4 hours of aeration.

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill
- Source: Hebron, Ohio
- Length at study initiation ( mean; range): 34 mm
- Weight at study initiation (mean; range): 0.64 g

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Hardness levels in the test vessels were not reported. The source water was reported to have a hardness of 103 mg/L (as CaCO3).

Test temperature:

22 degrees C in test vessels

pH:

7.9 - 9.1 reported in select test vessels; 7.6 in the source water

Dissolved oxygen:

Dissolved oxygen in the test vessels was not reported. Dissolved oxygen in the source water was reported as 7.7 mg/L.

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal concentrations not specified in the report.

Details on test conditions:

TEST SYSTEM
- Test vessel: 10 L round glass
- No. of organisms per vessel: 10
- Biomass loading rate: 0.64 g/L
Source/preparation of dilution water: declorinated Lake Huron water
- Metals:
Arsenic: <0.01 ppm
Barium: 0.006 ppm
Cadmium: nd
Chromium: <0.01 ppm
Copper: 0.02 ppm
Iron: 0.06 ppm
Lead: <0.01 ppm
Manganese: 0.002 ppm
Silver: nd
Zinc: <0.05 ppm
Mercury: nd
- Alkalinity: 85 mg/L
- Specific Conductance: 1770 micro mhos

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Death of fish every 24 hours

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

808 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Versenol 120

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI = 766-852

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

560 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Versenol 120

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

other: partial kill

Effect conc.:

750 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Versenol 120

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

other: 100% kill

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Versenol 120

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

750 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Versenol 120

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI = 665-786

Duration:

96 h

Dose descriptor:

other: LC90

Effect conc.:

870 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Versenol 120

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI = 830-980

Validity criteria fulfilled:

not applicable

Remarks:

no guideline available at the time of this study

Conclusions:

The 96 hour LC50 for bluegill sunfish was 808 mg/L Versenol 120.

Executive summary:

Bluegill sunfish were exposed to the test substance under static conditions at 22 degrees C for 96 hours and the NOEC, LC10, LC50, LC90, as well as partial kill and 100% kill were assessed. LC values were calculated using Finney's method of probit analysis. The LC50 was 808 mg/L, indicating relatively low toxicity to fish. There was limited test condition data reported (no dissolved oxygen levels reported for example). Also, the nominal concentrations that were included in the study design were not listed in the report. Analytical confirmation of the test substance was not performed.

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

The study with the read across substance is considered sufficient to fulfil the information requirements.

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

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

Key result

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

Key result

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

Key result

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:

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:

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:

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:

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:

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:

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.

Executive summary:

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.

Reference 6

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

supporting study

Study period:

30 September - 10 October 1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Some data on identity and purity, however no certificate of analysis and no actual concentrations. Limited phys-chem data available. Poor description of mat & meth., impossible to repeat, although basic essential info is given. Reliability 2 due to unclear natural water characteristics, illumination, >96 hours test concentration and oxygen and pH situation . Conclusions LC50 and NOEC up to 96 hours supportive.

Justification for type of information:

The study with the read across substance is considered sufficient to fulfil the information requirements.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

GLP compliance:

yes

Remarks:

In house QA

Analytical monitoring:

no

Details on sampling:

no chemical analyses

Vehicle:

not specified

Details on test solutions:

stock solution of 100 g/L was made in ultra pure water, pH 7.3
The dilution water was a natural water from the aquarium in which the fish were kept. This aquarium was purified by a biological filtration system
(including a mechanical filter, an aerobic sprinkling filter and an anaerobic nitrate removal (Minireef System)), containing water with the following
characteristics: pH 7.7, approximately 7 mg Ca and 20 mg Mg/l, 6ºdH.

concentrations: control, 0.1 - 0.18 - 0.32 - 0.56 and 1.0 g/L

Test solutions were renewed after 48 hours

Test organisms (species):

Poecilia reticulata

Details on test organisms:

The test organism was Poecilia reticulata (guppy); they were produced in the Akzo Research Laboratories Arnhem,
dept. CRL according to Standard Operation Procedures SOP CRL T 9 and T 42. The size was approximately 2 cm.
Loading of approx. 0.6 g/L

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Unclear, test duration 168 hours but not clear if fish were transferred to clean water or renewed test solutions. data only mention renewal once after 48 hours.

Hardness:

6ºdH

Test temperature:

24 ± 1ºC

pH:

7.3-7.9

Dissolved oxygen:

7.2 mg O2/L or higher

Salinity:

no data

Nominal and measured concentrations:

nominal concentrations: control, 0.1 - 0.18 - 0.32 - 0.56 and 1.0 g/L

Details on test conditions:

The temperature was kept between 23.5 and 24.5ºC, and the light regime w as 15 hours ambient light per day.
The loading in the test vessels was approximately 0.6 g biomass/l, 10 fish per test vessel, duplicate vessels per concentration.
Test was performed as a semi static test, water renewed after 48 hours. fish were approx. 2 cm in size.
No aeration, Open vessels, No food

Reference substance (positive control):

not specified

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

320 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Dissolvine Na2

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

180 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Dissolvine Na2

Basis for effect:

mortality (fish)

Details on results:

In the controls 1 out of 20 fish died within 96 hours. No explanation available.
A dose dependent mortality occurred in the treatment groups. 100% mortality was observed within 48 hours in the highest dose (1 g/L), within 73 hours in the 0.56 g/L groups and within 168 hours in the 0.32 g/L groups.
No mortality occurred in the 0.1 and 0.18 g/L groups.

Sublethal observations / clinical signs:

No. of surviving fish

Conc. In mg/L	0 h	24 h	48 h	73 h	96 h	124 h	168 h
Control	10	10	10	10	10	10	10
Control	10	10	10	10	9	9	9
100	10	10	10	10	10	10	10
100	10	10	10	10	10	10	10
180	10	10	10	10	10	10	10
180	10	10	10	10	10	10	10
320	10	10	10	8	4	1	0
320	10	10	10	9	7	3	0
560	10	10	2	0
560	10	10	1	0
1000	10	1	0
1000	10	1	0

Validity criteria fulfilled:

yes

Conclusions:

Guideline study done under GLP. However no chemical analyses and no certificate of analysis. In addition, there is limited info on identity and limited phys-chem properties. Mat & meth section poorly described lacking data on f.e. illumination, impossible to repeat. Data up to 96 hours can be considered supportive.

Executive summary:

A toxicity test was carried out with guppies (Poecilia reticulata) according to EEC (1984) C 1 in a semi static test. Dose range tested was 100 - 1000 mg/L with 20 fish per concentration, test solutions were renewed every 48 hours. Concentrations were tested in duplicate. In the controls 1 fish died within 96 h. In the 100 and 180 mg/L groups no fish died during the test (168h). The LC50-96h for guppy was 320 mg/l based on nominal concentrations, the NOEC based on mortality was concluded at nominally 180 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

372 mg/L

Marine water fish

Marine water fish

Effect concentration:

738 mg/L

Additional information

A number of studies have investigated the short-term toxicity to fish of trisodium HEDTA and the structural analogues disodium EDTA, tetrasodium EDTA, calcium disodium EDTA, manganese disodium EDTA and EDTA. As shown in Appendix I to the read across justification (see Section 13 of the IUCLID file), HEDTA, EDTA and their salts with non-toxic metals such as Na, Ca or Mg behave similarly in the aquatic environment and exert comparable toxicity to aquatic organisms. The toxicity is not linked to the structural difference between HEDTA and EDTA, nor to the counter-ion, but rather to factors such as water hardness, pH, presence of toxic/heavy metals and stoichiometry. The highest toxicity would be expected from uncomplexed HEDTA or EDTA is soft water, whereas HEDTA or EDTA metal chelates in hard water should present much lower toxicity. Indeed, the lowest 96 h LC50 value reported here is 41 mg H4-EDTA/L in bluegill sunfish (Lepomis macrochirus) in soft water whereas values of >1000 mg/L were found for MnNa2-EDTA in hard water (zebra fish, Danio rerio).

As data is available for Na3-HEDTA itself, these values were considered in priority for the present registration. The lowest 96 h LC50 is equivalent to 372 mg Versenol acid/L in fathead minnow (Pimephales promelas). Although water hardness is not given, this is the range of other data obtained for water of moderate hardness, which could be seen as more representative of a typical environmental situation than soft water. Therefore, the value of 372 mg/L was used for the CSA, representing both freshwater and marine fish. A study in a marine species (turbot, Scophtalmus maximus) exists with a 96 h LC50 of >738 mg Na3-HEDTA/L but this was not used for the CSA as no data on water hardness is available in the study and there is not enough information to conclude to lower toxicity for marine versus freshwater species.