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EC number: 268-006-8 | CAS number: 67989-22-4 This substance is identified in the Colour Index by Colour Index Constitution Number, C.I. 42535, molybdatephosphate salt.
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
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:
- supporting study
- Study period:
- April 2000
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that source and target substances have the same type of toxicological effects based on common underlying mechanisms (ECHA, 2015b). All substances have the same organic structural group (colorant) with different compositions of the inorganic group (metallic salt). Read-across is in this case based on the hypothesis that source and target substances have similar toxicological properties because they degrade to the similar chemical substances (ECHA, 2015c). This prediction is supported by physicochemical and toxicological data on the substances.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Name Source Substance Target Substance
Chemical name Benzenamine, N,N-dimethyl-, oxidized, molybdatetungstatephosphates Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatephosphates
IUPAC name reaction product of:
(a) [4-[bis[4-(dimethylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]-dimethyl-ammonium
(b) [4-[[4-(dimethylamino)phenyl]-[4-(methylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]-dimethyl-ammonium
(c) [4-[bis[4-(methylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]-dimethyl-ammonium
(d) phosphomolybdic acid
CAS No 101357-19-1 67989-22-4
EC No 309-916-8 268-006-8
Molecular weight range ≥ 2200 ≤ 3300 3800 - 7300
Molecular formula Reaction mass of C25 H30 N3. P Moy Wz O40 (y + z = 12), C24 H28 N3. P Moy Reaction mass of (C25H30N3)x.O3.P2O5.(MoO3)y; (C24H28N3)x.O3.P2O5.(MoO3)y and (C23H26N3)x.O3.P2O5.(MoO3)y
Wz O40 (y + z = 12) and C23 H26 N3. P Moy Wz O40 (y + z = 12) x = 6-10
y = 10-24
Structural formula
3. ANALOGUE APPROACH JUSTIFICATION
The target substance CAS 67989-22-4 is an organometallic multi-constituent substance. The source substance CAS 101357-19-1 has the same organic constituent (see table above) as the target substance consisting of triphenylmethan and three dimethylamine moieties. The inorganic Part B is very similar between source and target substances, consisting of the elements O, Mo, P and W in different compositions. These differences are expected to behave very similar. The target and source substances are thus considered suitable for the analogue approach based on structural similarity. - Qualifier:
- according to guideline
- Guideline:
- ISO 8692 (Water Quality - Fresh Water Algal Growth Inhibition Test with Scenedesmus subspicatus and Selenastrum capricornutum)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 0345N/982432
- Stability in water: at least 96hours - Analytical monitoring:
- no
- Details on sampling:
- - Concentrations: 0.1, 1, 10, 100mg/L in ISO-medium
- Vehicle:
- not specified
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: growing cultures of algae were exposed to concentrations of 0.1, 1, 10, 100mg/L in ISO-medium for 72 hours - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Pseudokirchnerialla subcapitata
- Strain: not specified
- Source (laboratory, culture collection): not specified - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Remarks on exposure duration:
- not specified
- Post exposure observation period:
- not specified
- Hardness:
- not specified
- Test temperature:
- controlled but not specified in report
- pH:
- controlled but not specified in report
- Dissolved oxygen:
- not specified
- Salinity:
- not specified
- Conductivity:
- not specified
- Nominal and measured concentrations:
- nominal
- Details on test conditions:
- TEST SYSTEM
- Initial cells density: 1 x 10e4 cells/mL
- Control end cells density: 147.2 x 10e4 cells/mL
- No. of vessels per concentration (replicates): triplicate
- No. of vessels per control (replicates):triplicate
GROWTH MEDIUM
- Standard medium used: yes
OTHER TEST CONDITIONS
- Sterile test conditions: yes/no
- Adjustment of pH:
- Photoperiod:
- Light intensity and quality:
- Salinity (for marine algae):
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: spectrophotometer
TEST CONCENTRATIONS
- Test concentrations: 0, 0.1, 1, 10, 100 mg/L
- Reference substance (positive control):
- no
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- < 0.1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- cell number
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- >= 0.1 - <= 1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Details on results:
- - Exponential growth in the control (for algal test): yes
- Algal cell growth was completeyl inhibited and the growth amd the growth rate completely reduced at 0345N concentrations prepared at 1mg/L and higher - Results with reference substance (positive control):
- none
- Reported statistics and error estimates:
- not specified
- Validity criteria fulfilled:
- yes
- Conclusions:
- The results show that a discharge of 0345N causes acute toxicity to algal growth
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- April 2000
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- ISO 8692 (Water Quality - Fresh Water Algal Growth Inhibition Test with Scenedesmus subspicatus and Selenastrum capricornutum)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 0345N/982432
- Stability in water: at least 96hours - Analytical monitoring:
- no
- Details on sampling:
- - Concentrations: 0.1, 1, 10, 100mg/L in ISO-medium
- Vehicle:
- not specified
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: growing cultures of algae were exposed to concentrations of 0.1, 1, 10, 100mg/L in ISO-medium for 72 hours - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Pseudokirchnerialla subcapitata
- Strain: not specified
- Source (laboratory, culture collection): not specified - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Remarks on exposure duration:
- not specified
- Post exposure observation period:
- not specified
- Hardness:
- not specified
- Test temperature:
- controlled but not specified in report
- pH:
- controlled but not specified in report
- Dissolved oxygen:
- not specified
- Salinity:
- not specified
- Conductivity:
- not specified
- Nominal and measured concentrations:
- nominal
- Details on test conditions:
- TEST SYSTEM
- Initial cells density: 1 x 10e4 cells/mL
- Control end cells density: 147.2 x 10e4 cells/mL
- No. of vessels per concentration (replicates): triplicate
- No. of vessels per control (replicates):triplicate
GROWTH MEDIUM
- Standard medium used: yes
OTHER TEST CONDITIONS
- Sterile test conditions: yes/no
- Adjustment of pH:
- Photoperiod:
- Light intensity and quality:
- Salinity (for marine algae):
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: spectrophotometer
TEST CONCENTRATIONS
- Test concentrations: 0, 0.1, 1, 10, 100 mg/L
- Reference substance (positive control):
- no
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- < 0.1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- cell number
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- >= 0.1 - <= 1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Details on results:
- - Exponential growth in the control (for algal test): yes
- Algal cell growth was completeyl inhibited and the growth amd the growth rate completely reduced at 0345N concentrations prepared at 1mg/L and higher - Results with reference substance (positive control):
- none
- Reported statistics and error estimates:
- not specified
- Validity criteria fulfilled:
- yes
- Conclusions:
- The results show that a discharge of 0345N causes acute toxicity to algal growth
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that source and target substances have the same type of toxicological effects based on common underlying mechanisms (ECHA, 2015b). All substances have the same organic structural group (colorant) with different compositions of the inorganic group (metallic salt). Read-across is in this case based on the hypothesis that source and target substances have similar toxicological properties because they degrade to the similar chemical substances (ECHA, 2015c). This prediction is supported by physicochemical and toxicological data on the substances.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Name Source Substance Target Substance
Chemical name Benzenamine, N,N-dimethyl-, oxidized, molybdatetungstatephosphates Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatephosphates
IUPAC name reaction product of:
(a) [4-[bis[4-(dimethylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]-dimethyl-ammonium
(b) [4-[[4-(dimethylamino)phenyl]-[4-(methylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]-dimethyl-ammonium
(c) [4-[bis[4-(methylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]-dimethyl-ammonium
(d) phosphomolybdic acid
CAS No 101357-19-1 67989-22-4
EC No 309-916-8 268-006-8
Molecular weight range ≥ 2200 ≤ 3300 3800 - 7300
Molecular formula Reaction mass of C25 H30 N3. P Moy Wz O40 (y + z = 12), C24 H28 N3. P Moy Reaction mass of (C25H30N3)x.O3.P2O5.(MoO3)y; (C24H28N3)x.O3.P2O5.(MoO3)y and (C23H26N3)x.O3.P2O5.(MoO3)y
Wz O40 (y + z = 12) and C23 H26 N3. P Moy Wz O40 (y + z = 12) x = 6-10
y = 10-24
Structural formula
3. ANALOGUE APPROACH JUSTIFICATION
The target substance CAS 67989-22-4 is an organometallic multi-constituent substance. The source substance CAS 101357-19-1 has the same organic constituent (see table above) as the target substance consisting of triphenylmethan and three dimethylamine moieties. The inorganic Part B is very similar between source and target substances, consisting of the elements O, Mo, P and W in different compositions. These differences are expected to behave very similar. The target and source substances are thus considered suitable for the analogue approach based on structural similarity. - Reason / purpose for cross-reference:
- read-across source
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch no 982432 supplied by Client
- Expiration date of the lot/batch: 01 July 2022
- Purity test date: Not supplied
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: poorly soluble
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not specified
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None
- Preliminary purification step (if any): None
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid: WAF'sprepared in Elendt M7 medium at nominal concentrations
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations:
- Sampling method: Not speicied
- Sample storage conditions before analysis:All samples were stored frozen prior to analysis. Duplicate samples were taken at each occasion and stored frozen for further analysis if necessary. - Vehicle:
- yes
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Nominal amounts of test item (10, 32, 20, 64 and 200 mg) were each separately added to the surface of 10, 10, 2.0, 2.0 and 2.0 liters of culture medium to give the 1.0, 3.2, 10, 32 and 100 mg/L loading rates respectively. After the addition of the test item, the culture medium was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 95 hours and the mixtures allowed to stand for 1-Hour. Visual observations made on the WAFs indicated that a significant amount of dispersed test item was present in the water column and hence it was considered justifiable to remove the WAF by filtering through a glass wool plug (2 to 4 cm in length). A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. A glass wool plug was inserted into the opposite end of the tubing and the WAF removed by mid-depth siphoning (the first 75 to 100 mL discarded). Microscopic observations of the WAFs were performed after filtering and showed no micro-dispersions of test item to be present. - Test organisms (species):
- other: Lemna gibba
- Details on test organisms:
- TEST ORGANISM
- Common name: Lemna gibba
- Strain: N/a
- Source (laboratory, culture collection): Culture obtained from Bayer CropScience AG, 40789 Monheim, Germany in 2007
- Age of inoculum (at test initiation):
- Method of cultivation:Maintained in the lab under standard test conditions
ACCLIMATION
- Acclimation period:7 days
- Culturing media and conditions (same as test or not): yes
- Any deformed or abnormal cells observed: no - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 7 d
- Test temperature:
- 24 ± 1 ºC throughout the test.
- pH:
- 5.6 – 5.7 on Day 0 to pH 6.4 – 6.5 on Day 7
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 500 mL glass conical flasks
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: fill volume 250 mL
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): N/a
- Renewal rate of test solution (frequency/flow rate): N/a
- Initial cells density:
- Control end cells density:
- No. of organisms per vessel: 3 colonies
- No. of vessels per concentration (replicates): three
- No. of vessels per control (replicates): three
- No. of vessels per vehicle control (replicates): n/a
GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used:
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
- Total organic carbon:
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:
- Ca/mg ratio:
- Conductivity:
- Culture medium different from test medium:
- Intervals of water quality measurement:
OTHER TEST CONDITIONS
- Sterile test conditions: yes/no
- Adjustment of pH:
- Photoperiod:
- Light intensity and quality:
- Salinity (for marine algae):
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter]
- Chlorophyll measurement:
- Other:
TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:
- Results used to determine the conditions for the definitive study: - Key result
- Duration:
- 7 d
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 7 d
- Dose descriptor:
- NOELR
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 7 d
- Dose descriptor:
- LOELR
- Effect conc.:
- 32 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 7 d
- Dose descriptor:
- EL50
- Effect conc.:
- 59 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Key result
- Duration:
- 7 d
- Dose descriptor:
- NOELR
- Effect conc.:
- 3.2 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Key result
- Duration:
- 7 d
- Dose descriptor:
- LOELR
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Validity criteria fulfilled:
- yes
- Conclusions:
- A study was performed to assess the effect of the test item on the growth of the freshwater plant Lemna gibba. Due to the low aqueous solubility and complex nature of the test item for the purposes of the test the test item was prepared as a Water Accommodated Fraction (WAF). Lemna gibba was exposed to WAFs of the test item over a range of nominal concentrations of 1.0, 3.2, 10, 32 and 100 mg/L loading rate WAF (three replicate flasks per concentration) for a period of 7 days, under constant illumination at a temperature of 24 ± 1 °C. The number of fronds in each control and treatment group was recorded on Days 0, 2, 4 and 7 along with observations on plant development.
Chemical analysis of the test preparations on Day 0 (fresh media) showed measured test concentrations to range from less than the limit of quantification (LOQ) of the analytical method, determined to be 0.048mg/L, to 0.39 mg/L. Analysis of the test preparations on Day 7 showed measured test concentrations of less than the LOQ were obtained from all test preparations.
The NOEL in terms of inhibition of yield calculated from dry weight was considered to be 3.2 mg/L loading rate WAF. Correspondingly the LOEL was considered to be 10 mg/L loading rate WAF. - Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 08 February 2018 to 28 March 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline No. 221 “Lemna sp. Growth Inhibition Test (March 2006)”
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch no 982432 supplied by Client
- Expiration date of the lot/batch: 01 July 2022
- Purity test date: Not supplied
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: poorly soluble
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not specified
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None
- Preliminary purification step (if any): None
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid: WAF'sprepared in Elendt M7 medium at nominal concentrations
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations:
- Sampling method: Not speicied
- Sample storage conditions before analysis:All samples were stored frozen prior to analysis. Duplicate samples were taken at each occasion and stored frozen for further analysis if necessary. - Vehicle:
- yes
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Nominal amounts of test item (10, 32, 20, 64 and 200 mg) were each separately added to the surface of 10, 10, 2.0, 2.0 and 2.0 liters of culture medium to give the 1.0, 3.2, 10, 32 and 100 mg/L loading rates respectively. After the addition of the test item, the culture medium was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 95 hours and the mixtures allowed to stand for 1-Hour. Visual observations made on the WAFs indicated that a significant amount of dispersed test item was present in the water column and hence it was considered justifiable to remove the WAF by filtering through a glass wool plug (2 to 4 cm in length). A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. A glass wool plug was inserted into the opposite end of the tubing and the WAF removed by mid-depth siphoning (the first 75 to 100 mL discarded). Microscopic observations of the WAFs were performed after filtering and showed no micro-dispersions of test item to be present. - Test organisms (species):
- other: Lemna gibba
- Details on test organisms:
- TEST ORGANISM
- Common name: Lemna gibba
- Strain: N/a
- Source (laboratory, culture collection): Culture obtained from Bayer CropScience AG, 40789 Monheim, Germany in 2007
- Age of inoculum (at test initiation):
- Method of cultivation:Maintained in the lab under standard test conditions
ACCLIMATION
- Acclimation period:7 days
- Culturing media and conditions (same as test or not): yes
- Any deformed or abnormal cells observed: no - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 7 d
- Test temperature:
- 24 ± 1 ºC throughout the test.
- pH:
- 5.6 – 5.7 on Day 0 to pH 6.4 – 6.5 on Day 7
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 500 mL glass conical flasks
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: fill volume 250 mL
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): N/a
- Renewal rate of test solution (frequency/flow rate): N/a
- Initial cells density:
- Control end cells density:
- No. of organisms per vessel: 3 colonies
- No. of vessels per concentration (replicates): three
- No. of vessels per control (replicates): three
- No. of vessels per vehicle control (replicates): n/a
GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used:
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
- Total organic carbon:
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:
- Ca/mg ratio:
- Conductivity:
- Culture medium different from test medium:
- Intervals of water quality measurement:
OTHER TEST CONDITIONS
- Sterile test conditions: yes/no
- Adjustment of pH:
- Photoperiod:
- Light intensity and quality:
- Salinity (for marine algae):
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter]
- Chlorophyll measurement:
- Other:
TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:
- Results used to determine the conditions for the definitive study: - Key result
- Duration:
- 7 d
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 7 d
- Dose descriptor:
- NOELR
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 7 d
- Dose descriptor:
- LOELR
- Effect conc.:
- 32 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 7 d
- Dose descriptor:
- EL50
- Effect conc.:
- 59 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Key result
- Duration:
- 7 d
- Dose descriptor:
- NOELR
- Effect conc.:
- 3.2 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Key result
- Duration:
- 7 d
- Dose descriptor:
- LOELR
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Validity criteria fulfilled:
- yes
- Conclusions:
- A study was performed to assess the effect of the test item on the growth of the freshwater plant Lemna gibba. Due to the low aqueous solubility and complex nature of the test item for the purposes of the test the test item was prepared as a Water Accommodated Fraction (WAF). Lemna gibba was exposed to WAFs of the test item over a range of nominal concentrations of 1.0, 3.2, 10, 32 and 100 mg/L loading rate WAF (three replicate flasks per concentration) for a period of 7 days, under constant illumination at a temperature of 24 ± 1 °C. The number of fronds in each control and treatment group was recorded on Days 0, 2, 4 and 7 along with observations on plant development.
Chemical analysis of the test preparations on Day 0 (fresh media) showed measured test concentrations to range from less than the limit of quantification (LOQ) of the analytical method, determined to be 0.048mg/L, to 0.39 mg/L. Analysis of the test preparations on Day 7 showed measured test concentrations of less than the LOQ were obtained from all test preparations.
The NOEL in terms of inhibition of yield calculated from dry weight was considered to be 3.2 mg/L loading rate WAF. Correspondingly the LOEL was considered to be 10 mg/L loading rate WAF.
Referenceopen allclose all
Description of key information
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 100 mg/L
- EC10 or NOEC for freshwater algae:
- 10 mg/L
Additional information
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