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EC number: 276-911-4 | CAS number: 72829-25-5
- 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 plants other than algae
Administrative data
Link to relevant study record(s)
- Endpoint:
- toxicity to aquatic plants other than algae
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 06 April 2016 to 17 May 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 221 (Lemna sp. Growth Inhibition Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- UK GLP
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: MC-9
- Expiration date of the lot/batch: 11 May 2020
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Stored frozen at approximately -20 ºC in the dark - Analytical monitoring:
- not specified
- Details on sampling:
- Samples were taken from the control and each test group from the bulk test preparation on Day 0 and from the pooled replicates on Day 7 for quantitative 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
- Remarks:
- The culture medium used for both the range-finding and definitive tests was the same as that used to maintain the stock culture.
- Details on test solutions:
- Culture Medium
NaNO3 85 mg/L
KH2PO4 13.4 mg/L
MgSO4.7H2O 75 mg/L
CaCl2.2H2O 36 mg/L
Na2CO3 20 mg/L
H3BO3 1.0 mg/L
MnCl2.4H2O 0.20 mg/L
Na2MoO4.2H2O 0.010 mg/L
ZnSO4.7H2O 0.050 mg/L
CuSO4.5H2O 0.0050 mg/L
Co(NO3)2.6H2O 0.010 mg/L
FeCl3.6H2O 0.84 mg/L
Na2-EDTA.2H2O 1.40 mg/L
The culture medium will be prepared in reverse osmosis purified water (Elga Optima 15+ or Elga Purlab Option R-15 BP). The pH of the prepared culture medium will be adjusted, if necessary, to 6.5 ± 0.2 with either 1M HCl or NaOH. The culture was maintained in the laboratory at a temperature of 24 ± 1 ºC under continuous illumination (intensity approximately 7000 lux) for at least 7 days prior to the start of the test. - Test organisms (species):
- Lemna minor
- Details on test organisms:
- The test was carried out using Lemna minor. A culture of Lemna minor was obtained from Canadian Phycological Culture Centre, University of Waterloo, Ontario, Canada. Cultures were maintained in the laboratory by the periodic replenishment of culture medium.
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 7 d
- Hardness:
- None
- Test temperature:
- 24 ± 1 ºC
- pH:
- Day 0: 6.6 to 6.7
Day 7: 7.3 to 7.6 - Nominal and measured concentrations:
- Range-finding Test:
1.0, 10 and 100 mg/L
Definitive Test
Based on the results of the range-finding test the following test concentrations were assigned to the definitive test: 6.25, 12.5, 25, 50 and 100 mg/L. - Details on test conditions:
- Range-finding Test
The test concentrations to be used in the definitive test were determined by a preliminary range-finding test. The range-finding test was conducted by exposing Lemna minor to a series of nominal test concentrations of 1.0, 10 and 100 mg/L for a period of 7 days. The test was conducted in glass conical flasks (500 mL). Two replicate flasks were prepared for each control and test concentration. The test item was dissolved directly in culture medium. A nominal amount of test item (100 mg) was dissolved in culture medium and the volume adjusted to 1 liter to give a 100 mg/L test concentration from which a series of dilutions was made to give further test concentrations of 10 and 1.0 mg/L. Each of the prepared concentrations was inverted several times to ensure adequate mixing and homogeneity. The control group was maintained under identical conditions but not exposed to the test item. At the start of the range-finding test the number of fronds present in each test and control culture was recorded along with observations on frond size, appearance, root length and number of colonies present. The flasks were then incubated at 24 ± 1 ºC under continuous illumination (intensity approximately 7000 lux) for 7 days. On Days 2 and 5 the test solutions were renewed, and observations on the test organisms were recorded on days 0, 2, 5 and 7. In order to determine the stability of the test item under test conditions a sample of each test concentration was taken for chemical analysis on Day 2 (fresh media) and Day 5 (old media). An additional sample of each test concentration was prepared on Day 0 and incubated alongside the test until Day 7 in order to determine stability over the entire test duration. All samples were stored frozen prior to analysis.
Definitive Test
Based on the results of the range-finding test the following test concentrations were assigned to the definitive test: 6.25, 12.5, 25, 50 and 100 mg/L.
Experimental Preparation
A nominal amount of test item (200 mg) was dissolved in culture medium and the volume adjusted to 2 liters to give a 100 mg/L test concentration from which a series of dilutions was made to give further test concentrations of 50, 25, 12.5 and 6.25 mg/L. Each of the prepared concentrations was inverted several times to ensure adequate mixing and homogeneity. The concentration and stability of the test item in the test preparations were verified by chemical analysis on Day 0 and Day 7.
Exposure Conditions
As in the range-finding test glass conical flasks were used. Three flasks each containing 250 mL of solution were prepared for the control and each treatment group. The control group was maintained under identical conditions but not exposed to the test item. Each control and test flask was inoculated with 3 colonies of Lemna minor (total 11 fronds). The flasks were then incubated at 24 ± 1 ºC under constant illumination (intensity approximately 7000 lux) for 7 days. A static testing regime was employed.
Assessments
Test Organism Observations
The number of fronds present in each test and control culture was recorded on days 0, 3, 5 and 7 along with observations on frond size, appearance, root length and number of colonies present. In addition, the dry weight of the fronds in each control and treatment group was determined on day 7. At the start of the test six replicate samples of fronds identical to those used to inoculate the test vessels were taken and the dry weight determined. At the end of the test the dry weight of colonies from each control and test vessel was determined by blotting the colonies dry and drying at 60 °C to constant weight.
Water Quality Criteria
The pH of each control and test flask was recorded on Day 0 and Day 7. The temperature and light intensity in the incubator were recorded daily.
Verification of Test Concentrations
Samples were taken from the control and each test group from the bulk test preparation on Day 0 and from the pooled replicates on Day 7 for quantitative analysis. All samples were stored frozen prior to analysis. Duplicate samples were taken at each occasion and stored frozen for further analysis if necessary.
Data Evaluation
Doubling Time
The doubling time (Td) of frond number, and hence performance of the test against the validation criterion of doubling time, was calculated for the control vessels.
Response Variables
In order that the test results are acceptable to regulatory authorities worldwide, the effects on the growth of Lemna were evaluated using both response variables (a) and (b) described below:
a) Average specific growth rate: this response variable was calculated on the basis of changes in the logarithms of frond numbers, and in addition, on the basis of changes in the logarithms of dry weight over time (expressed per day) in the controls and each treatment group.
b) Yield: this response variable was calculated on the basis of changes in frond number, and in addition, on the basis of changes in dry weight in the controls and in each treatment group until the end of the test. It should be noted that toxicity values calculated using these two response variables are not comparable and the difference must be recognized when using the results of the test. ECx values based upon average specific growth rate (ErCx) will be higher than results based upon yield (EyCx) under the test conditions of this Guideline, due to the underlying mathematics of the respective approaches. This is not to be interpreted as a difference in sensitivity between the response variables: simply the values are different mathematically. The concept of average specific growth rate is based upon the general exponential growth pattern of Lemna in non-limited cultures. Toxicity is estimated on the basis of the effects on the growth rate without being dependent on the absolute level of the specific growth rate of the control, slope of the concentration-response curve or on test duration. Results based upon yield are dependent upon all these variables.
Average Specific Growth Rate
The average specific growth rate for a specific period was calculated as the logarithmic increase in the growth variables -frond numbers and dry weight - for each replicate of the control and treatment groups. For each control and treatment group the mean value for growth rate along with the standard deviation was calculated.
Yield
Effects on yield were determined on the basis of frond numbers and dry weight present in each test vessel at the start and end of the test. For dry weight, the starting biomass was determined on the basis of a sample of fronds taken from the same batch used to inoculate the test vessels. For each test concentration and control, the mean value for yield along with variance estimates was calculated.
Determination of ECx Values
Percentage inhibition (arithmetic axis) was plotted against test concentration (logarithmic axis) and a line fitted by computerized interpolation using the Xlfit software package (IDBS). ECx values were then determined from the equation for the fitted line.
It was not possible to calculate 95 % confidence limits for the EC50 values as the data generated did not fit the models available for the calculation of confidence limits.
Statistical analysis
One way analysis of variance incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf, 1981), and Dunnett's multiple comparison procedure for comparing several treatments with a control (Dunnett, 1955), were carried out on the average specific growth rate and yield data at 7 days for the control and all test concentrations to determine any statistically significant differences between the test and control groups. All statistical analyses were performed using the SAS computer software package (SAS, 1999-2001).
Validation Criteria
The results of the test are considered valid if the following performance criterion is met:
The doubling time of frond numbers in the controls must be less than 2.5 days (60 h), corresponding to approximately a seven-fold increase in seven days and an average specific growth rate of 0.275 d-1. - Reference substance (positive control):
- yes
- Remarks:
- A positive control (Envigo Study Number MM01PC) used 3,5-dichlorophenol as the reference item. Details of the positive control are given in Annex 1. The positive control was conducted between 18 May 2016 and 30 May 2016.
- Key result
- Duration:
- 7 d
- Dose descriptor:
- EC50
- 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:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 7 d
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 7 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 7 d
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Duration:
- 7 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Duration:
- 7 d
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Duration:
- 7 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Details on results:
- Range-finding Test
The results showed no significant effect on growth at the test concentration of 1.0 and 10 mg/L. However, growth was observed to be reduced at 100 mg/L. Based on this information test concentrations of 6.25, 12.5, 25, 50 and 100 mg/L were selected for the definitive test. Chemical analysis of the 10 and 100 mg/L test preparations on Days 2, 5 and 7 showed near nominal concentrations were obtained indicating that the test item was stable under test conditions.
Definitive Test
Verification of Test Concentrations
Chemical analysis of the test preparations on Day 0 (fresh media) and Day 7 (old media) showed measured test concentrations to range from 94 % to 103 % of nominal and so the results are based on nominal test concentrations only.
Validation Criteria
The following data show that the doubling time of the control cultures was 1.83 days in line with the OECD Guideline that states the doubling time should be less than 2.5 days: Mean frond number in control cultures at day 0: 11 Mean frond number in control cultures at day 7: 104
Growth Data Based on Frond Number
Accordingly, the following results based on inhibition of average specific growth rate and yield were determined from the frond number data:
Average Specific Growth Rate
ErC10 (frond number) = 38 mg/L
ErC20 (frond number) ≥100 mg/L
ErC50 (frond number) ≥100 mg/L
Where:
ErCx = the test concentration that reduced average specific growth rate by x%.
Statistical analysis of the average specific growth rate data was carried out for the control and all test concentrations using one way analysis of variance incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf 1981) and Dunnett's multiple comparison procedure for comparing several treatments with a control (Dunnett 1955). There were no statistically significant differences between the control, 6.25, 12.5, 25, 50 and 100 mg/L test concentrations (P≥0.05) and therefore the "No Observed Effect Concentration" (NOEC) in terms of inhibition of average specific growth rates calculated from frond numbers was 100 mg/L.
Yield
EyC10 (frond number) = 8.9 mg/L
EyC20 (frond number) = 40 mg/L
EyC50 (frond number) ≥100 mg/L
Where:
EyCx = the test concentration that reduced yield by x%.
Statistical analysis of the yield data was carried out for the control and all test concentrations. There were no statistically significant differences between the control, 6.25, 12.5, 25, 50 and 100 mg/L test concentrations (P≥0.05) and therefore the "No Observed Effect Concentration" (NOEC) in terms of inhibition of yield calculated from frond numbers was 100 mg/L.
Growth Data Based on Dry Weight
Accordingly the following results based on inhibition of average specific growth rate and yield were determined from the dry weight data:
Average Specific Growth Rate
ErC10 (dry weight) = 99 mg/L
ErC20 (dry weight) ≥100 mg/L
ErC50 (dry weight) ≥100 mg/L
Where:
ErCx = the test concentration that reduced average specific growth rate by x%.
Statistical analysis of the average specific growth rate data was carried out for the control and all test concentrations. There were no statistically significant differences between the control 6.25, 12.5, 25, 50 and 100 mg/L test concentrations (P≥0.05) and therefore the "No Observed Effect Concentration" (NOEC) in terms of inhibition of average specific growth rate calculated from dry weight was 100 mg/L.
6.2.8 Yield
EyC10 (dry weight) = <6.25 mg/L EyC20 (dry weight) = 9.2 mg/L EyC50 (dry weight) = >100 mg/L
Where:
EyCx = the test concentration that reduced yield by x%.
Statistical analysis of the yield data was carried out for the control and all test concentrations as in Section 6.2.4. There were no statistically significant differences between the control, 6.25, 12.5, 25, 50 and 100 mg/L test concentrations (P≥0.05) and therefore the "No Observed Effect Concentration" (NOEC) in terms of inhibition of yield calculated from dry weight was 100 mg/L.
Observations
All test and control cultures were inspected on days 0, 3, 5 and 7.
Water Quality Criteria
Temperature was maintained at 24 ± 1 ºC throughout the test. - Results with reference substance (positive control):
- A positive control (Envigo Study Number MM01PC) used 3,5-dichlorophenol as the reference item at concentrations of 0.625, 1.25, 2.5, 5.0 and 10 mg/L. Exposure conditions and data evaluation for the positive control were similar to those in the definitive test.
- Validity criteria fulfilled:
- yes
- Conclusions:
- The EC50 and NOEC for growth rate of frond number as well as dry weight was >100 and 100 mg/L, respectively.
- Executive summary:
The effect of the test item on the growth of the freshwater plant Lemna minor was evaluated in a study conducted according to the OECD Guideline 221. Following a preliminary range-finding test, Lemna minor was exposed to an aqueous solution of the test item at concentrations of 6.25, 12.5, 25, 50 and 100 mg/L (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, 3, 5 and 7 along with observations on plant development. Chemical analysis of the test preparations on Day 0 (fresh media) and Day 7 (old media) showed measured test concentrations to range from 94 % to 103 % of nominal and so the results are based on nominal test concentrations only. Lemna gibba exposed to FAT 40034 recorded: 7 day ErC50 based upon the frond number is >100 mg/L. 7 day EyC50 based upon the frond numbers is >100 mg/L. 7 day ErC50 based upon the frond dry weight is >100 mg/L. 7 day EyC50 based upon the frond dry weight is >100 mg/L. Based on the findings of the study, the EC50 and NOEC of FAT 40034/F for growth rate of frond number as well as dry weight was >100 and 100 mg/L, respectively.
Reference
Exposure of Lemna minor to the reference item gave the following results:
Response Variable |
Measurement Variable |
EC50(mg/L) |
95% Confidence Limits |
No Observed Effect Concentration (NOEC) (mg/L) |
Lowest Observed Effect Concentration (LOEC) (mg/L) |
||
Average Specific Growth Rate |
Frond Number |
3.4 |
3.1 |
- |
3.8 |
0.625 |
1.25 |
Dry Weight |
3.0 |
2.7 |
- |
3.2 |
0.625 |
1.25 |
|
Yield |
Frond Number |
1.8 |
1.6 |
- |
2.2 |
0.625 |
1.25 |
Dry Weight |
1.4 |
1.2 |
- |
1.7 |
0.625 |
1.25 |
The results from the positive control with 3,5-dichlorophenol were within the normal ranges for this reference item.
Description of key information
The effect of the test item on the growth of the freshwater plant Lemna minor was evaluated in a study conducted according to the OECD Guideline 221. Following a preliminary range-finding test, Lemna minor was exposed to an aqueous solution of the test item at concentrations of 6.25, 12.5, 25, 50 and 100 mg/L (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, 3, 5 and 7 along with observations on plant development. Chemical analysis of the test preparations on Day 0 (fresh media) and Day 7 (old media) showed measured test concentrations to range from 94 % to 103 % of nominal and so the results are based on nominal test concentrations only. Lemna gibba exposed to FAT 40034 recorded: 7 day ErC50 based upon the frond number is >100 mg/L. 7 day EyC50 based upon the frond numbers is >100 mg/L. 7 day ErC50 based upon the frond dry weight is >100 mg/L. 7 day EyC50 based upon the frond dry weight is >100 mg/L. Based on the findings of the study, the EC50 and NOEC of FAT 40034/F for growth rate of frond number as well as dry weight was >100 and 100 mg/L, respectively.
Key value for chemical safety assessment
- EC50 for freshwater plants:
- 100 mg/L
- EC10 or NOEC for freshwater plants:
- 100 mg/L
Additional information
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