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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
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:
Daphnia magna were exposed for 48 hours in a static system at 17°C to the test substance.
GLP compliance:
no
Analytical monitoring:
no
Details on sampling:
Not reported
Vehicle:
not specified
Details on test solutions:
Not reported
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain: rafinesque
- All other template details: Not reported
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
Not reported
Hardness:
Not reported
Test temperature:
17°C
pH:
Not reported
Dissolved oxygen:
Not reported
Salinity:
Not reported
Nominal and measured concentrations:
Nominal (mg/L): 135, 240
Details on test conditions:
Not reported
Reference substance (positive control):
not specified
Key result
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
192 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Versenol acid
Basis for effect:
mortality
Remarks on result:
other: 95% CI: 187-197 mg/L
Remarks:
Water hardness not reported
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
< 135 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
other: partial kill
Effect conc.:
135 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC100
Effect conc.:
240 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
Not reported
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
Thompson's method of moving averages was used to calculate 48 hour LC50 values.

Not reported

Conclusions:
The 48 hours LC50 for Daphnia magna for Versenol acid chelating agent was 192 mg/L.
Executive summary:

Daphnia magna were exposed to the test substance under static conditions at 17 degrees C for 48 hours and the NOEC, LC50, as well as partial kill and 100% kill was assessed. LC Values were calculated using Thompson's method of moving averages. The LC50 was 192 mg/L, indicating relatively low toxicity to daphnids. There was limited test condition data reported. Analytical confirmation of the test substance was not performed.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27 to 29 October 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
equivalent or similar to
Guideline:
other: Thompson 1990 - Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea)
Deviations:
not specified
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
no
Details on sampling:
Not applicable
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: test concentrations were prepared by addition of specified quantities of the sample to dilution water
- Dilution water: natural seawater filtered to 5 µm and supplied by Guernsey Sea Farms. Stored in the dark at about 15 degrees C. Immediately before use, the water was aerated until the dissolved oxygen reached saturation and the pH stabilized at 8.0 +/- 0.3
- Evidence of undissolved material: none, a clear colorless solution was obtained
Test organisms (species):
other aquatic crustacea: Acartia tonsa
Details on test organisms:
TEST ORGANISM - Acartia tonsa
- Source: Guernsey Sea Farms
- Age at study initiation: adult
- Feeding during test: Not reported

ACCLIMATION
- Feeding frequency: every other day
- Health during acclimation (any mortality observed): not reported

QUARANTINE: Not applicable
Test type:
static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
Not reported
Hardness:
Not reported
Test temperature:
20.5 degrees C
pH:
control: 8.0
5600 mg/L exposure: 9.9 initially, 8.9 ending
Dissolved oxygen:
control: 91 - 93 %ASV
5600 mg/L exposure: 95-96 %ASV
Salinity:
38-40%
Nominal and measured concentrations:
Nominal (mg/L): 0, 560, 1000, 1800, 3200, 5600
Details on test conditions:
TEST SYSTEM
- Test vessel: 50 mL crystallising dishes
- Material, size, headspace, fill volume: filled with 25 ml of dilution water
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per positive control (replicates): 4
- Biomass loading rate: not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: natural sea water supplied by Guernsey Sea Farms
- No water parameters reported

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

EFFECT PARAMETERS MEASURED: mortality observed at 24 and 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: roughly 2x
- Range finding study
- Test concentrations: not reported
- Results used to determine the conditions for the definitive study: LC50 greater than 1000 mg/L
Reference substance (positive control):
yes
Remarks:
3,5 Dichlorophenol
Key result
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 5 600 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Versenol/120 Chel
Basis for effect:
mortality
Remarks on result:
other:
Remarks:
Water hardness not reported
Key result
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 2 296 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
Na3-HEDTA
Basis for effect:
mortality
Remarks on result:
other:
Remarks:
Water hardness not reported
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
1 800 mg/L
Nominal / measured:
estimated
Basis for effect:
mortality
Remarks on result:
other: determined by Fisher's exact test
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
738 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
at 41%
Basis for effect:
mortality
Remarks on result:
other: determined by Fisher's exact test
Details on results:
- Mortality of control: None
- All other template details: Not reported
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Mortality: 40% mortality at 48 hours
- EC50/LC50: not calculated
- Other:
Reported statistics and error estimates:
Where possible, the LC50 values were estimated and 95% confidence limits calculated using ToxCalc version 5 "Comprehensive Toxicity Data Analysis and Database Software"

Table 1. Cumulative percent mortalities.

 Nominal concentration (mg/L)  Number Dead     Percent Mortality   
   24 hrs 48 hrs  24 hrs  48 hrs
 0  0  0  0  0
 560  0  0  0  0
 1000  0  0  0  0
 1800  0  1  0  5
 3200  2  5  10  25
 5600  2  8  10  40
 1 mg/L 3,5 Dichlorophenol  1  8  5  40
Validity criteria fulfilled:
yes
Conclusions:
Versenol/120 Chel was of low toxicity to Acartia tonsa, only 40% mortality was recorded at the highest test concentration of 2296 mg/L after 48 hours. The extrapolated 48 hour LC50 was 2585 mg/L as determined by the Maximum Likelihood-Probit method.
Executive summary:

Versenol/120 Chel was of low toxicity to Acartia tonsa, only 40% mortality was recorded at the highest test concentration of 2296 mg/L after 48 hours. The extrapolated 48 hour LC50 was 2585 mg/L as determined by the Maximum Likelihood-Probit method.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1989 June 14-16
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
other: annex V of Policy 79/831 EWG
Deviations:
not specified
Principles of method if other than guideline:
Daphnia magna were exposed in 10 mL containers, 4 replicates with 5 animals per replicate, to the test substance in freshwater for 48 hours for effects on mobility.
GLP compliance:
not specified
Analytical monitoring:
no
Details on sampling:
Not reported
Vehicle:
not specified
Details on test solutions:
Test material was added directly to dilution water.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain: Daphnia magna Strauss
- Source: National Institute of Applied Chemical Research, France
- Age at study initiation: 2-24 hrs
- All other template details: not reported

ACCLIMATION: Not reported

QUARANTINE (wild caught): Not applicable
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
Not reported
Hardness:
2.70 +/- 0.5 mmol/L
Test temperature:
292-294 K
pH:
8.0 +/- 0.5
Dissolved oxygen:
8.01 - 9.12 mg/L
Salinity:
Not reported
Nominal and measured concentrations:
Nominal (mg/L): 0, 62.5, 125, 250, 500
Details on test conditions:
TEST SYSTEM
- Test vessel: centrifuge jars with a flattened bottom filled with 10 ml of test water
- Aeration: the test water is up to the oxygen saturation of ventilated air, then left to stand for 24 hours for stabilization.
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- All other template details: not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: See Table 1
- Ca/Mg ratio: 4:1
- Na/K ratio: 10:1
- Conductivity: 600-700 µSiemens/cm
- Intervals of water quality measurement: before study

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

EFFECT PARAMETERS MEASURED: mortality observed at 0, 3, 6, 24, and 48 hrs by ability to swim

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- All other template details: Not reported
Reference substance (positive control):
not specified
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 500 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Trilon D
Basis for effect:
mobility
Remarks on result:
other: no effect at highest concentration tested
Remarks:
Hard water
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
>= 500 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: no effect at highest concentration tested
Details on results:
- Results: all 20 animals per test concentration were alive and mobile at each observation period
- Behavioural abnormalities: none observed
- Mortality of control: 0/20 animals
- Other adverse effects control: none observed
Results with reference substance (positive control):
Not reported
Reported statistics and error estimates:
Not applicable (no effects seen at highest concentration tested)

Not applicable

Conclusions:
No effects were seen at any of the test concentrations (0, 62.5, 125, 250, 500 mg/L) of the test substance - Trilon D - to Daphnia magna exposed in static water for 48 hours.
Executive summary:

Daphnia magna were exposed to the test substance - Trilon D- for 48 hours and observed for mobility/mortality. There were no effects seen at the highest concentration tested, 500 mg/L.

Endpoint:
short-term toxicity to aquatic invertebrates
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:
test procedure in accordance with national standard methods with acceptable restrictions
Justification for type of information:
The study with the read across substance is considered sufficient to fulfil the information requirements.
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
other: DIN 38412, part 11
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
no
Test organisms (species):
Daphnia magna
Details on test organisms:
Species: Daphnia magna
Origin: wild population
Breeding: The clone with the highest number of offspring was used to establish a continuous daphnia breed under standardized laboratory conditions.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
24 h
Key result
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
625 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Na4-EDTA
Basis for effect:
mobility
Remarks on result:
other: Water hardness not reported
Duration:
24 h
Dose descriptor:
EC0
Effect conc.:
310 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC100
Effect conc.:
1 250 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
In this short-term test, Ca was present in overstoichiometric amounts. Thus, in the test system the Ca-complex was formed.
Endpoint:
short-term toxicity to aquatic invertebrates
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:
test procedure in accordance with national standard methods
Justification for type of information:
The study with the read across substance is considered sufficient to fulfil the information requirements.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
no
Test organisms (species):
Daphnia magna
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
24 h
Key result
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
610 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Na4-EDTA
Basis for effect:
mobility
Remarks on result:
other: Water hardness not reported
EC50(24h) calculated (confidence interval 95 %):
----------------------------------------------------
EDTA 610 mg/l (570-640)

Mn(II)+EDTA 940 mg/l (800-1000)
Fe(III)+EDTA 17 mg/l (13-21)
Cu(II)+EDTA 38 mg/l (31-45)
Zn(II)+EDTA 910 mg/l (840-980)
Hg(II)+EDTA 0.00032 mg/l (0.00022-0.00052)
Cd(II)+EDTA 310 mg/l (290-330)
----------------------------------------------------

All the EC50 values, except those obtained for the free EDTA, are expressed as mg/L of total amount of the studied metal. To compare the toxicities of the metal chelates with those of the free EDTA, all the EC50 values were converted to mg/L EDTA.
EC 50 values with 95% confidence limits were calculated by a probit  analysis computer program, version 2.3 (National Swedish Environmental Protection Agency).

The EC50 values of Fe3+, Cu2+, Mn2+, Zn2+, Cd2+ and Hg2+ complexed by EDTA were compared with those of the respective uncomplexed metals and free EDTA.
The experiments gave the following toxicity order: Hg2+, Cu2+, Cd2+, Zn2+, Fe3+, Mn2+ (higher to lower toxicity).
After complexation, the toxicities were 1.1-1700 (EDTA) times lower. With Hg2+ complexation did not decrease the toxicity. Compared with the toxicity of free EDTA, metal complexation increased the toxicity of EDTA.

Description of key information

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
192 mg/L

Additional information

A number of studies have investigated the short-term toxicity to aquatic invertebrates of trisodium HEDTA and the structural analogues disodium EDTA and tetrasodium EDTA. The 24/48 h EC50 values range from 192 mg Versenol acid/L in Daphnia magna to >2296 mg Na3-HEDTA/L in the marine copepod Acartia tonsa.The lowest value for Na3-HEDTA of 192 mg Versenol acid/L in Daphnia magna was considered for the CSA. Although water hardness is not given, this is the range of other data obtained in fish for water of moderate hardness, which could be seen as more representative of a typical environmental situation than soft water. A study in a marine species exists 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.