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EC number: 217-895-0 | CAS number: 2001-94-7
Toxicity to aquatic algae and cyanobacteria
Administrative data
Link to relevant study record(s)
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Guideline study
- 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
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- yes
- Remarks:
- modified test system without iron limitation (medium differing to OECD)
- Principles of method if other than guideline:
- The study was conducted according to the EU guideline EEC/79/831, Annex V, C, Algae and the standard operating procedures (SOP) of the BASF laboratory.
- GLP compliance:
- no
- Analytical monitoring:
- no
- Vehicle:
- no
- Details on test solutions:
- Stock solutions:
249.75 mg/L (Na4-EDTA)
177.55 mg/L (FeCl3 x 6 H20) - Test organisms (species):
- Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
- Details on test organisms:
- Test species: Scenedesmus subspicatus
Strain: CHODAT SAG 86.81
Source: Algae collection Goettingen, SAG 86.81 - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Test temperature:
- 23 +/- 2 °C
- pH:
- ca. 8
- Nominal and measured concentrations:
- Test concentrations: 263.04 µmol/L - 1.03 µmol/L
- Details on test conditions:
- TEST CONDITIONS
Illumination: 120 µEinstein/m*m*s (permanent Illumination, Color of light was universal white, L 25 (Osram))
Inoculation density: 10000 cells/mL (initial concentration in the test)
TEST SYSTEM
Aeration: none
No. of replicates:
3 inoculated replicates
1 uninoculated replicate for measuring the pH-value of the uninoculated test concentrations after an incubation time of 72 h.
Controls: negative control
TEST MEDIUM / WATER PARAMETERS
Dilution water: OECD medium differing to the SOP of the laboratory
Each test concentration was mixed according to the following regimen:
1 part algal cell suspension in OECD medium
1 part OECD medium 9-times concentrated
4 parts FeCl3 x 6 H20 dilutions in H20
4 parts Na4-EDTA dilutions in H20 - Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC90
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Details on results:
- The validity criteria were fulfilled.
Factor of the cell proliferation (cell count) after 72 h: 36.7 - Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Study period:
- 22-7-2009 - 25-7-2009
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: This study was conducted to the OECD guidelines with a minor deviation (see below). No analysis were performed. The screening study concerned was inaccordance with OECD guidelines but was not GLP audited. Control validity criteria were met.
- 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
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Deviations:
- not specified
- Principles of method if other than guideline:
- The NaHCO3 concentration of the test medium was 150 mg/l in stead of 50 mg/l, in order to maintain a more constant pH during the test.
- GLP compliance:
- no
- Remarks:
- GLP accredited laboratory not conducted to GLP
- Analytical monitoring:
- no
- Vehicle:
- no
- Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Remarks on exposure duration:
- This study was conducted as a series of preliminary tests for EDTA for metal complexes.
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 307.63 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 1 000 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 1 000 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Validity criteria fulfilled:
- yes
- Conclusions:
- This study can be concluded reliable with restrictions.
- Executive summary:
The study concerned was in accordance with OECD guidelines in a GLP accredited laboratory, but was not GLP audited.
No chemical analysis was performed. Control growth was satisfactory and current reference test with potassium
dichromate had been conducted on the algae species used for testing. This study can be considered reliable with the
restrictions mentioned. And is therefore reliable as supporting evidence.
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Study period:
- 2001-03-19 to 2001-04-06
- Reliability:
- 1 (reliable without restriction)
- 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
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Version / remarks:
- 1984
- Deviations:
- no
- GLP compliance:
- yes
- Analytical monitoring:
- yes
- Details on sampling:
- Samples of about 3 mL were taken from 3 control vessels and from each test vessels containing test substance at 0 24,48 and 72 hours. The samples were stored in the freezer (<-10 degree C) until analyses.
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
A stock solution/suspension of the test substance was prepared as follows: an accurately measured amount of 0.6526 g of test substance was dissolved in deionized water. Exactiy 4.0 ml of a 118.5 g/L FeCI3x6H20 was added with a pipette to reach an equimolar concentration of EDTA and Fe(lll). After stirring the pH of the solution was adjusted to pH 5.2 using a 1 M NaOH Solution. The final concentration of 5.1237 g/l based on H4EDTA was reached by adding deionized water to a final volume of 100 ml. The stock Solution was stored in the refrigerator in the dark.
The test solutions were prepared by addition of the required amounts of stock Solution to the test vessels to obtain the following final nominal test concentrations: 60, 80 and 100 mg/l. - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: P.subcapitata
- Strain: CCAP 278/4
- Source (laboratory, culture collection): Culture Collection of Algae and Protozoa, Institute of Freshwater Ecology, The Windermere Laboratory, Cumbria, Ambleside, United Kingdom. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Test temperature:
- 23 +/- 2 degree C
- Salinity:
- Not applicable
- Nominal and measured concentrations:
- Nominal concentrations of 60, 80 and 100 mg/L
- Details on test conditions:
- TEST SYSTEM
- Test vessel: Erlenmeyer test flasks
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume: 100 mL, closed with cotton-wool stoppers
GROWTH MEDIUM
- Standard medium used: yes
OTHER TEST CONDITIONS
- Adjustment of pH: jes
- Light intensity and quality: continuous uniform illumination was provided in the spectral ränge of 400 to 700 nm by using 30 W fluorescent lamps
EFFECT PARAMETERS MEASURED:
- Determination of cell concentrations: photometrically with a UVA/IS Spectrophotometer - Reference substance (positive control):
- no
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 60 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 79.4 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 48.4 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 99.9 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 60.6 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Validity criteria fulfilled:
- yes
- Conclusions:
- In conclusion the 72 hour EC50 value for Fe(III)EDTA with P.subcapitata was greater than 100 mg/L based on nominal concentrations and greater than 60 mg/L based on the mean measured concentrations. The NOEC was 79.4 mg/L based on nominal concentrations and 48.4 mg/L based on the mean measured concentrations.
- Executive summary:
The toxicity of Fe(lll)EDTA to exponentially growing Pseudokirchneriella subcapitata was determined over an exposure period of 72 hours. A test was carried out at nominal concentrations of 60, 80 and 100 mg/L Fe (lll) EDTA (concentrations represented as H4EDTA).Due to photodegradation, the Fe(lll)EDTA concentration declines during the test. The chemical analyses carried out showed that the concentration of the test compound at the end of the test had decreased to about 50% of the concentration at the beginning of the test. The mean concentrations during the 72 hours of testing for the nominal concentrations of 60, 80 and 100 mg/L as measured by chemical analyses were 38.1, 48.4 and 60.6 mg/L, respectively. The test was conducted in a mineral salt medium in a climatized illuminated orbital incubator. The maximum variation in pH in the test media was 1.4 pH unit. In conclusion the 72 hour EC50 value for Fe(III)EDTA with P.subcapitata was greater than 100 mg/L based on nominal concentrations and greater than 60 mg/L based on the mean measured concentrations. The NOEC was 79.4 mg/L based on nominal concentrations and 48.4 mg/L based on the mean measured concentrations.
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Guideline study conducted in compliance with GLP regulations
- 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
- Qualifier:
- according to guideline
- Guideline:
- other: EEC guideline 79/831/EEC, Annexe V, part C: Methods for the determination of ecotoxicity, algae: Test of growth inhibition, May 1988
- GLP compliance:
- yes
- Analytical monitoring:
- yes
- Test organisms (species):
- Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
- Details on test organisms:
- Test species: Scenedesmus subspicatus
Strain: CHODAT SAG 86.81
Source: Algae collection Goettingen, SAG 86.81 - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Test temperature:
- 23 +/- 2 °C
- pH:
- 7.65 - 8.80
- Details on test conditions:
- Aeration: none
No. of replicates:
3 inoculated replicates
1 uninoculated replicate for measuring the pH-value of the uninoculated test concentrations after an incubation time of 72 h.
Controls: negative control
TEST MEDIUM / WATER PARAMETERS
Dilution water: according to EEC guideline 79/831/EEC, Annexe V, part C:
Methods for the determination of ecotoxicity, algae: Test of growth inhibition, May 1988 and the SOP of the laboratory
Deviation from the SOP: Each test concentration was mixed according to the following regimen:
1 part of inoculum (1E+05 cells/ml in test medium)
1 part of test medium (10 times concentrated)
8 parts of test substance (in appropriate water dilutions)
TEST CONDITIONS
Illumination: 120 uEinstein/m*m*s (permanent Illumination, Color of light was universal white, L 25 (Osram))
Inoculation density: 10000 cells/mL (initial concentration in the test)
TEST CONCENTRATIONS
Test concentrations: 100 mg/l - 0.39 mg/L
Stock solution: 125 mg/L - Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.39 mg/L
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 0.78 mg/L
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 0.7 mg/L
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 2.77 mg/L
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC90
- Effect conc.:
- >= 100 mg/L
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
Referenceopen allclose all
Effect on Biomass:
------------------
EbC 10 = > 263.04 umol/l (> 100 mg/l Na4-EDTA)
EbC 50 = > 263.04 umol/l (> 100 mg/l Na4-EDTA)
EbC 90 = > 263.04 umol/l (> 100 mg/l Na4-EDTA)
Effect on Growth rate:
----------------------
ErC 10 = > 263.04 umol/l (> 100 mg/l Na4-EDTA)
ErC 50 = > 263.04 umol/l (> 100 mg/l Na4-EDTA)
ErC 90 = > 263.04 umol/l (> 100 mg/l Na4-EDTA)
NOEC (72 h) = 263.04 umol/l (100 mg/l Na4-EDTA)
LOEC (72 h) = > 263.04 umol/l (> 100 mg/l Na4-EDTA)
Observations:
-------------
Over the whole range of test concentrations, a stimulation of the cell proliferation was observed (max. 30.4 % based on biomass).
This study was conducted as a series of preliminary tests for EDTA for metal complexes.
| Concentration | Time (hours) | Area | Inhibition | Specific | Inhibition | |
| (mg/L) | 0 | 72 | Percentage | Growth Rate | Percentage | |
| 0 | 0.007 | 0.501 | 17.784 | 0.059315222 | ||
| 0 | 0.007 | 0.492 | 17.46 | 0.059063452 | ||
| 0 | 0.006 | 0.35 | 12.384 | 0.056474635 | ||
| 0 | 0.008 | 0.285 | 9.972 | 0.049625662 | ||
| 0 | 0.007 | 0.464 | 16.452 | 0.058249644 | ||
| 0 | 0.008 | 0.265 | 9.252 | 0.048615115 | ||
| Mean | 0.007167 | 0.3928333 | 13.884 | 0.055610344 | 0 | |
| 1 | 0.006 | 0.377 | 13.356 | 0.055081866 | ||
| 1 | 0.006 | 0.45 | 15.984 | 0.05753328 | ||
| 0 | 0 | |||||
| - | - | |||||
| Mean | 0.006 | 0.4135 | 14.67 | 56.611.927 | 0.056361588 | 13.509.065 |
| 10 | 0.008 | 0.453 | 16.02 | 0.057564031 | ||
| 10 | 0.008 | 0.374 | 13.176 | 0.054897007 | ||
| 0 | ||||||
| Mean | 0.008 | 0.4135 | 14.598 | 5.1426102 | 0.056294438 | 1.2301551 |
| 100 | 0.025 | 0.365 | 12.24 | 0.052977253 | ||
| 100 | 0.024 | 0.563 | 19.404 | 0.061179004 | ||
| 0 | ||||||
| Mean | 0.0245 | 0.464 | 15.822 | 13.958513 | 0.057394055 | -3.207516 |
| 1000 | 0 | 0.032 | 1.152 | 0.028630870 | ||
| 1000 | 0.005 | 0.116 | 3.996 | 0.035060394 | ||
| 0 | 0 | |||||
| Mean | 0.0025 | 0.074 | 2.574 | 81.4606742 | 0.033274707 | 40.1645378 |
Effect on the development of biomass [mg/L]:
EbC 10 = 0.48 mg/L
EbC 50 = 1.01 mg/L
EbC 90 = 100 mg/L
Effect on growth rate [mg/L]:
ErC 10 = 0.70 mg/L
ErC 50 = 2.77 mg/L
ErC 90 => 100.0 mg/L
No observed effect concentration (95 % significance level):
NOEC (72 h) = 0.39 mg/L
Lowest observed effect concentration:
LOEC (72 h) = 0.78 mg/L
VALIDTY CRITERIA:
The validity criteria were maintained.
Factor of cell division (cell count) after 72 hours: 46
SUMMARY OF THE CONCENTRATION CONTROL ANALYSIS:
The concentrations 100 and 12.5 mg/L, the untreated control and the stock solution (125 mg/L) were analysed. Test
conditions and results are listed in the GLP-reports 94LOO650 and 94LOO663, dated 22 Nov 1994, from Dr. Scholtissek, BASF AG, ZAX/OF.
Time scan:
begin of test (0 h) - uninoculated replicate
end of test (72 h) - uninoculated replicate
end of test (72 h) - inoculated replicate after removal of algal cells through centrifugation
Recovery in %:
control / test / stock solution /
-----------------------------------------------------------
t= 0 h uninoculated <DL / 100-104 / 99.2 /
t= 72 h uninoculated <DL / 96-99 / 98.4 /
t= 72 h inoculated <DL / 99-104 / - /
-----------------------------------------------------------
DL= detection limit (1 mg/L)
Description of key information
With high probability the test substance is acutely not harmful to aquatic algae.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 300 mg/L
Additional information
For the hazard assessment on green algae several guideline compliant studies have been performed using Na4-EDTA, FeNa -EDTA and Fe(III)EDTA, for justification for read-across see IUCLID 5, Chapter 13.
According to BASF 94/1080/60/1 an inhibitory effect of the test substance was shown, which was caused by complexing essential micronutrients from the culture medium. Therefore algae growth can be significantly reduced under standardized guideline conditions and may yield apparent results that are more or less severe than the true toxicity (see Guidance on Hazard to the Aquatic Environment Globally Harmonized System GHS Annex 9 A9.3.3.4, 2007). To compensate this secondary effect, the algae study with Scenedesmus subspicatus was performed once in nutrient- enriched media with FeCl3 (BASF, 95/9999/60/1). Finally the EC50 (72h) for the growth rate was determined to be nominal > 100 mg/L. as no analytical dose verification was performed. The 72 hour EC50 value for Fe(III)EDTA with P.subcapitata was greater than 100 mg/L based on nominal concentrations and greater than 60 mg/L based on the mean measured concentrations (Akzo Nobel 2001). Further on, the toxicity of FeNa-EDTA was tested in standard OECD medium. The calculated EC50 of nominal > 1000 mg/L FeNa-EDTA were based on range finding test with a broad range (Akzo Nobel 2009).
The results of these four studies indicates that the predicted LC50 for EDTA (acid form) and its salts are higher than measured 60 mg/L and can be estimated to be higher than 300 mg/L, which is in line with the EU Risk Assessment (2004).
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
