Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 268-028-8 | CAS number: 67990-05-0
- 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 soil macroorganisms except arthropods
Administrative data
Link to relevant study record(s)
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2009-06-10 to 2009-08-06
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study compliant to GLP
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
- Version / remarks:
- DIN ISO 11268-2: 1998 (E), BBA Guideline (1994)
- Deviations:
- yes
- Remarks:
- Room temperature increased once to about 22.5 °C for four hours. The soil moisture of all test vessels deviated by more than 10 % of the initial value at the end of the test.
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- GLP according to German Chemikaliengesetz and Directive 88/320/EEC
- Analytical monitoring:
- no
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Method of mixing into soil (if used): The respective test item concentration was weighed out, dissolved in acetone (2.5 mL per replicate) and mixed thoroughly with quartz sand (10 g per replicate). After complete evaporation of the solvent, the treated quartz sand was given to the artificial soil. Demineralised water was added to adjust the humidity of the artificial soil to a moisture of 52 % of the maximum water holding capacity. Subsequently, the test medium will be mixed with a mixer for at least three minutes and 600 g soil dry weight were filled into each test vessel.
- Controls: Artificial soil with addition of quartz sand moistened with demineralised water without test item.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): organic solvent: acetone
- Concentration of vehicle in test medium (stock solution and final test solution): 2.5 mL acetone per replicate, 10 g quartz sand per replicate
- Evaporation of vehicle before use: Yes - Test organisms (species):
- Eisenia fetida
- Animal group:
- annelids
- Details on test organisms:
- TEST ORGANISM
- Common name: Eisenia fetida SAVIGNY 1826, subspecies Eisenia fetida fetida
- Source: Own breeding
- Age at test initiation (mean and range, SD): synchronized earthworm culture, not deviating more than 1 month
- Weight at test initiation (mean and range, SD): 0.44 +/- 0.05 g
ACCLIMATION
- Acclimation period: 2 days prior to test start
- Acclimation conditions (same as test or not): same as test
- Health during acclimation (any mortality observed): healthy - Study type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- no
- Total exposure duration:
- 56 d
- Remarks:
- Removement of adults after 28 days
- Test temperature:
- 18 - 22.5 deg C
- pH:
- Application rate
[mg test item/kg soil DW] pH-value
day 0 day 56
Control 5.42 6.13
Solv. Control 5.40 6.40
62.5 5.39 6.18
125 5.39 6.30
250 5.40 6.33
500 5.38 6.07
1000 5.42 6.14 - Moisture:
- Application rate
[mg test item/kg soil DW] Moisture [%] of dry weight
day 0 day 56
Control 35.6 42.5*
Solv. Control 35.1 45.8*
62.5 34.8 46.7*
125 35.5 46.8*
250 34.9 44.6*
500 35.2 45.2*
1000 34.6 44.6*
*Variation of the water content > 10 %, this deviation is considered to have no impact on the study - Details on test conditions:
- TEST SYSTEM
- Test container (material, size): round plastic boxes, ID: 15 cm, bottom surface area: 177 cm3, height: 14 cm, transparent and perforated lids enabled sufficient gas exchange and light penetration and prevented the test substrate from drying
- Amount of soil or substrate: 600 g DW
- No. of organisms per container (treatment): 10 (40)
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 4
SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10 % peat, 20 % kaolin, 69 % air dried quartz sand, 0.4 % calcium carbonate
- Organic carbon (%): not determined
- Maximum water holding capacity (in % dry weigth): 68
- CEC: not determined
- Pretreatment of soil: none
- Storage (condition, duration): 8 days after preparation at test conditions in the dark
OTHER TEST CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 524 +/- 70 lx
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Adult mortality, biomass, behaviour and pathological symptoms after 28 days
Reproduction (Number of juveniles) after 56 days
VEHICLE CONTROL PERFORMED: yes
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 - Nominal and measured concentrations:
- Nominal application rates: 62.5 - 125 - 250 - 500 - 1000 mg test substance/kg soil dry weight (DW)
- Reference substance (positive control):
- yes
- Remarks:
- 1.0 - 2.5 - 5.0 mg Carbendazim/ kg soil DW
- Duration:
- 56 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1 000 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- reduction of body weight, mortality of adult earthworms
- Details on results:
- - Mortality at end of exposure period: No mortality of adult earthworms was observed in the controls as well as at all application rates after 28 days of exposure to the test item
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: At test start the earthworms had an individual weight of 0.30 - 0.60 g. The body weights of the earthworms increased in the control and at the test item concentrations of 62.5, 125, 250, 500 and 1000 mg test item / kg soil dry weight throughout the first 28 days. The biomass did not differ statistically significant compared to the pooled control at all test item concentrations
- No. of offspring produced: The mean number of juveniles in the pooled control was 359. The average number of juveniles in the treated groups ranged between 288 and 365. Compared to the pooled control the reproduction of the earthworms did not statistically significantly differ at all test item application rates
- Behavioural abnormalities: No symptoms or changes in the behaviour of the earthworms were observed in the controls as well as at all application rates after 28 days of exposure to the test item - Results with reference substance (positive control):
- - Results with reference substance valid? Yes
- Relevant effect levels: 2.5 mg Carbendazim/kg soil DW - Reported statistics and error estimates:
- Differences between the control and the solvent control were statistically analysed using t-tests. No statistical differences occurred between the controls concerning biomass and reproduction. Thus, both controls were pooled for further statistical analyses.
One Way Analysis of Variance (ANOVA) was carried out for the determination of statistically significant differences compared to
the control. A Normality Test and an Equal Variance Test were conducted prior to running the ANOVA. P-values for both Normality and Equal Variance Test are 0.05. The a-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) is a = 0.05. - Validity criteria fulfilled:
- yes
- Conclusions:
- In this earthworm reproduction test according to OECD 222 the submission substance did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentrations of 62.5, 125, 250, 500 and 1000 mg/kg soil dry weight for 28 days. Also, the biomass increase of the adult earthworms after 28 days did not statistically significantly differ compared to the pooled control at all test item concentrations. In addition, after eight weeks of exposure the earthworm reproduction was not statistically significantly reduced at all applied test item concentrations.
Thus the No Observed Effect Concentration (NOEC) for the submission substance with respect to mortality, body weight and reproduction of Eisenia fetida fetida after 8 weeks of exposure in artificial soil was determined to be 1000 mg test item/kg soil dry weight. - Executive summary:
Effects of the submission substance on mortality, biomass and the reproductive potential of the earthworm species Eisenia fetida (Savigny) were determined compliant to GLP according to the guidelines DIN ISO 11268-2 (1998), OECD 222 (2004) and BBA (1994) (reliability category 1). The study was conducted under static conditions over 8 weeks with five different application rates (62.5 - 125 - 250 - 500 - 1000 mg test item/kg soil dry weight), which were mixed into artificial soil containing 10 % peat. 40 test organisms, with an individual weight of 0.30 to 0.60 g, were divided into 4 replicate groups with 10 test organisms per replicate.
After 28 days no mortality as well as no pathological symptoms or changes in the behaviour of adult earthworms were seen in the control, in the solvent control and at all tested application rates. The earthworm biomass increased in the control and at all application rates over 28 days without any statistically significant differences compared to the control. After further four weeks the reproduction rate (average number of juveniles) was 359 in the control (control and solvent control) and ranged between 288 and 365 at the test item application rates. Compared to the control, the reproduction rate did not statistically significantly differ at all application rates.
Overall, the NOEC of the submission substance concerning mortality, biomass and reproduction of earthworms was determined to be >= 1000 mg test item/kg soil dry weight (Table 2).
All validity criteria recommended by the test guidelines were fulfilled.
Summary of All Observed Effects in the Tested Doses
Effects
Test item
mg test item/kg soil dry weight
62.5
125
250
500
1000
Mortality of adult earthworms
No
No
No
No
No
Reduction of body weight
No
No
No
No
No
Reduced reproduction rate
(Number of juveniles)
No
No
No
No
No
Summary of all effects
No
No
No
No
No
NOEC and LOEC for Reproduction, Mortality and Body Weight
Test item
Application rate
mg test item/kg soil dry weight
NOECreproduction, mortality, body weight
1000
LOECreproduction, mortality, body weight
> 1000
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2008-11-21 to 2009-01-16
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline-study compliant to GLP
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
- Version / remarks:
- DIN ISO 11268-2 and BBA Guideline
- Deviations:
- yes
- Remarks:
- The room temperature decreased three times to about 17.5 °C and increased three times to about 21 °C . The soil moisture of all test vessels deviated by more than 10 %. Deviations considered to have no impact on study quality and integrity.
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- GLP according to German Chemikaliengesetzt and Directive 88/320/EEC
- Analytical monitoring:
- no
- Details on sampling:
- No analytical monitoring
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Method of mixing into soil (if used): The respective test item concentration was weighed out, dissolved in acetone (2.5 mL per replicate) and mixed thoroughly with quartz sand (10 g per replicate). After complete evaporation of the solvent, the treated quartz sand was given to the artificial soil. Demineralised water was added to adjust the humidity of the artificial soil to a moisture of 52 % of the maximum water holding capacity.
Subsequently, the test medium was mixed with a long handled blender for at least three minutes and filled into the test vessels.
- Controls: Artificial soil with addition of quartz sand moistened with demineralised water without test item was used as control medium.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Quartz sand treated with acetone
- Concentration of vehicle in test medium (stock solution and final test solution): 2.5 mL per replicate
- Evaporation of vehicle before use: Yes - Test organisms (species):
- Eisenia fetida
- Animal group:
- annelids
- Details on test organisms:
- TEST ORGANISM
- Common name: Eisenia fetida SAVIGNY 1826, subspecies Eisenia fetida fetida
- Source: Own breeding
- Age at test initiation (mean and range, SD): synchronized earthworm culture, not deviating more than 1 month
- Weight at test initiation (mean and range, SD): 0.54 +/- 0.04 g
ACCLIMATION
- Acclimation period: 2 days prior to test start
- Acclimation conditions (same as test or not): same as test
- Health during acclimation (any mortality observed): healthy - Study type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- no
- Total exposure duration:
- 56 d
- Post exposure observation period:
- No
- Test temperature:
- Room Temperature [°C]
nominal 20 +/- 2
actual 17.5 - 21.5*
*) The room temperature decreased three times to about 17.5 deg C for a total of 9 hours and increased three times to about 21 deg C for a total of 12 hours. - pH:
- Application rate
[mg test item/kg soil DW] pH-value
day 0 day 56
Control 6.07 6.42
Solv. Control 5.99 6.32
62.5 5.71 6.51
125 5.71 6.65
250 5.71 6.58
500 5.82 6.55
1000 5.69 6.73 - Moisture:
- Application rate
[mg test item/kg soil DW] Moisture [%] of dry weight
day 0 day 56
Control 33.5 38.3*
Solv. Control 32.3 39.4*
62.5 32.5 39.9*
125 32.5 39.2*
250 32.4 39.3*
500 32.4 37.2*
1000 31.2 38.9*
*) Variation of the water content > 10 %, this deviation is considered to have no impact on the study. - Details on test conditions:
- TEST SYSTEM
- Test container (material, size): round plastic boxes, ID: 15 cm, bottom surface area: 177 cm3, height: 14 cm, transparent and perforated lids enabled sufficient gas exchange and light penetration and prevented the test substrate from drying
- Amount of soil or substrate: 600 g DW
- No. of organisms per container (treatment): 10 (40)
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10 % peat, 20 % kaolin, 69 % air dried quartz sand, 0.36 % calcium carbonate
- Organic carbon (%): not determined
- Maximum water holding capacity (in % dry weigth): 65
- CEC: not determined
- Pretreatment of soil: none
- Storage (condition, duration): 9 days after preparation at test conditions in the dark
OTHER TEST CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 614 ± 67.7 lx
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Adult mortality, biomass, behaviour and pathological symptoms after 28 days
Reproduction (Number of juveniles) after 56 days
VEHICLE CONTROL PERFORMED: yes
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 - Nominal and measured concentrations:
- nominal concentrations: 62.5 - 125 - 250 - 500 - 1000 mg test item/kg soil DW
- Reference substance (positive control):
- yes
- Duration:
- 56 d
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 1 000 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- mortality and reduction of body weight
- Details on results:
- - Mortality at end of exposure period: No mortality was observed in any of the tested test item and control groups
- Total mass of adults at beginning of test: 0.49 - 0.50
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: Control and solvent control: 46.8 +/- 7.47, 62.5 mg/kg soil DW: 46.0 +/- 2.09, 125 mg/kg soil DW: 50.7 +/- 11.3, 250 mg/kg soil DW: 57.1 +/- 8.37, 500 mg/kg soil DW: 56.4 +/- 4.73, 1000 mg/kg soil DW: 57.9 +/- 2.50
- No. of offspring produced: Control AND SOLVENT CONTROL: 235 +/- 39.1, 62.5 mg/kg soil DW: 244 +/- 30.4, 125 mg/kg soil DW: 238 +/- 61.6, 250 mg/kg soil DW: 269 +/- 42.6, 500 mg/kg soil DW: 237 +/-67.5, 1000 mg/kg soil DW: 267 +/- 31.4
- No. of unhatched cocoons: Not determined
- Morphological abnormalities: None
- Behavioural abnormalities: None
- Other biological observations: None - Results with reference substance (positive control):
- - Results with reference substance valid? Yes
Reference Item (Carben) - Reproduction Rate (Number of Juveniles after 8 weeks)
Application rate
[mg a.i./kg soil dry weight] Rep. Number of juveniles Mean ± SD CV Reduction compared to control [%] Significance*
Control 1 248 272
±
34.8 12.8 - -
2 236
3 305
4 298
1.0 1 224 259
±
47.4 18.3 4.60 No
2 263
3 225
4 325
2.5 1 39 44.8
±
13.8 30.9 83.5 Yes
2 65
3 34
4 41
5.0 1 0 0
±
0.0 - 100 Yes
2 0
3 0
4 0
Rep. = Replicate a.i. = Active ingredient SD = Standard deviation CV = Coefficient of variation - Reported statistics and error estimates:
- Differences between the control and the solvent control were statistically analysed using One Way Analysis of Variance (ANOVA). No statistical differences occurred between the controls concerning biomass and reproduction. Thus, both controls were pooled for further statistical analyses.
One Way Analysis of Variance (ANOVA) was carried out for the determination of statistically significant differences compared to
the control. A Normality Test and an Equal Variance Test were conducted prior to running the ANOVA. P-values for both Normality and Equal Variance Test are 0.05. The a-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) is a = 0.05. Since the Normality Test failed using reproduction data, the data were log transformed for ANOVA to obtain a passed Normality Test.
The EC50-value for reproduction was not calculated since no reduction of reproduction occurred at all application rates. - Validity criteria fulfilled:
- yes
- Conclusions:
- Under the conditions of this study, the submission substance did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentrations of 62.5, 125, 250, 500 and 1000 mg/kg soil dry weight for 28 days. Also, the biomass increase of the adult earthworms after 28 days did not statistically significantly differ compared to the pooled control at all test item concentrations. In addition, after 56 days of exposure the earthworm reproduction was not statistically significantly reduced at all applied test item concentrations.
Thus the No Observed Effect Concentration (NOEC) for the submission substance with respect to mortality, body weight and reproduction of Eisenia fetida fetida after 8 weeks of exposure in artificial soil was determined to be >= 1000 mg test item/kg soil dry weight. - Executive summary:
Effects of the submission substance on mortality, biomass and the reproductive potential of the earthworm species Eisenia fetida (Savigny) were determined according to the guidelines DIN ISO 11268-2 (1998), OECD 222 (2004) and BBA (1994) compliant to GLP (reliability category 1). The study was conducted under static conditions over 8 weeks with five different application rates (62.5, 125, 250, 500, and 1000 mg test item/kg soil dry weight), which were mixed into artificial soil containing 10 % peat. 40 test organisms, with an individual weight of 0.30 to 0.60 g, were divided into 4 replicate groups with 10 test organisms per replicate.
After 28 days no mortality as well as no pathological symptoms or changes in the behaviour of adult earthworms were seen in the control, in the solvent control and at all tested application rates (Table 1). The earthworm biomass increased in the control and at all application rates over 28 days without any statistically significant differences compared to the control. After further four weeks the reproduction rate (average number of juveniles) was 235 in the control (control and solvent control) and ranged between 237 and 269 at the test item application rates. Compared to the control, the reproduction rate did not statistically significantly differ at all application rates.
Overall, the NOEC of Pigment Red 187 concerning mortality, biomass and reproduction of earthworms was determined to be >= 1000 mg test item/kg soil dry weight.
All validity criteria recommended by the test guidelines were fulfilled.
Summary of All Observed Effects in the Tested Doses
Effects
Test Item
mg test item/kg soil dry weight
62.5
125
250
500
1000
Mortality of adult earthworms
No
No
No
No
No
Reduction of body weight
No
No
No
No
No
Reduced reproduction rate
(Number of juveniles)
No
No
No
No
No
Summary of all effects
No
No
No
No
No
NOEC and LOEC for Reproduction, Mortality and Body Weight
Test Item
Application rate
mg test item/kg soil dry weight
NOECreproduction, mortality, body weight
>= 1000
LOECreproduction, mortality, body weight
> 1000
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2012-03-15 to 2012-05-11
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Performed according to relevant guidelines and compliant to GLP
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
- Deviations:
- yes
- Remarks:
- Deviations are decribed in the field 'Principles of method if other than guideline'.
- Principles of method if other than guideline:
- Food was provided on day 0 instead of day 1. This deviation is considered to have no impact on quality and integrity of the study.
During 1 day the room temperature increased to approx 23°C. During 1 day the room temperature decreased below 18°C. These deviations are considered to have no impact on the quality and integrity of the study.
Variation of the soil water content slightly > 10%: This deviation is considered to have no impact on the quality and integrity of the study. - GLP compliance:
- yes (incl. QA statement)
- Remarks:
- GLP according to German Chemikaliengesetzt and Directive 88/320/EEC
- Analytical monitoring:
- no
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Method of mixing into soil (if used): The respective test item amount was weighed out for the limit test item concentration, dissolved in acetone (2.5 mL per replicate) and mixed thoroughly with quartz sand (10 g per replicate). After complete evaporation of the solvent, the treated quartz sand was given to the artificial soil. Additional demineralised water was added to the artificial soil to adjust the humidity of the artificial soil to a moisture of 52 % of the maximum water holding capacity.
Subsequently, the test medium was thoroughly mixed to ensure a homogeneous distribution and about 600 g soil dry weight were filled into each test vessel.
- Controls: Artificial soil moistened with demineralised water without test or reference item and without solvent was used as control medium.
- Solvent control: Artificial soil with addition of quartz sand treated with acetone and moistened with demineralised water without test item was used as solvent control. - Test organisms (species):
- Eisenia fetida
- Animal group:
- annelids
- Details on test organisms:
- TEST ORGANISM
- Common name: Eisenia fetida SAVIGNY 1826, subspecies Eisenia fetida fetida
- Source: Own breeding
- Age at test initiation (mean and range, SD): synchronized earthworm culture, not deviating more than 1 month
- Weight at test initiation (mean and range): 0.44 g, 0.3-0.6 g
ACCLIMATION
- Acclimation period: 2 days prior to test start
- Acclimation conditions (same as test or not): same as test
- Health during acclimation (any mortality observed): healthy - Study type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- yes
- Total exposure duration:
- 56 d
- Remarks:
- Removal of adults after 28 days
- Test temperature:
- 16* - 23*
*) During 1 day the room temperature increased to approx 23°C. During 1 day the room temperature decreased
below 18°C. These deviations are considered to have no impact on the quality and integrity of the study. For the remaining days temperature was at 20 +/- 2 deg C. - pH:
- Application rate
[mg / kg soil dry weight] pH-value
day 0 day 56
Control 5.71 6.35
Solvent Control 5.71 6.27
1000 5.53 6.17 - Moisture:
- Application rate
[mg / kg soil dry weight] Moisture [%] of dry weight
day 0 day 56
Control 33.1 37.4*
Solvent Control 32.9 37.1*
1000 33.2 36.8*
*) Variation of the water content slightly > 10%: This deviation is considered to have no impact on the quality and integrity of the study. - Details on test conditions:
- TEST SYSTEM
- Test container (material, size): round plastic boxes, ID: 15 cm, bottom surface area: 177 cm3, height: 14 cm, transparent and perforated lids enabled sufficient gas exchange and light penetration and prevented the test substrate from drying
- Amount of soil or substrate: 600 g DW
- No. of organisms per container (treatment): 10 (80)
- No. of replicates per treatment group: 8
- No. of replicates per control: 8
SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10 % peat, 20 % kaolin, 69 % air dried quartz sand, 0.41 % calcium carbonate
- Organic carbon (%): not determined
- Maximum water holding capacity (in % dry weigth): 65
- CEC: not determined
- Pretreatment of soil: none
- Storage (condition, duration): 8 days after preparation at test conditions in the dark
OTHER TEST CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 548 +/- 83 lx
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Adult mortality, biomass, behaviour and pathological symptoms after 28 days
Reproduction (Number of juveniles) after 56 days
VEHICLE CONTROL PERFORMED: Yes
- Nominal and measured concentrations:
- Nominal application rate: 1000 mg test substance/kg soil dry weight (DW)
- Reference substance (positive control):
- yes
- Remarks:
- 1.0 - 2.5 - 5.0 mg Carbendazim/ kg soil DW
- Duration:
- 56 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1 000 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- , mortality, body weight
- Details on results:
- - Mortality at end of exposure period:
No mortality of adult earthworms in the controls and at the limit test item concentration was observed after 28 days of exposure to the test item
- Changes in body weigth of live adults (% of initial weight) at end of exposure period:
At test start the earthworms had an individual weight of 0.37 - 0.54 g. As there were no statistically significant differences determined between the control and the solvent control for the body weight, the controls were pooled. The body weights of the adult earthworms increased throughout the first 28 days at the limit application rate with no statistically significant differences compared to the pooled control.
- No. of offspring produced:
As there were no statistically significant differences determined between the control and the solvent control for the reproduction rate, the controls were pooled.
The mean number of juveniles in the pooled control was 185. The average number of juveniles in the treated group was 164. Compared to the pooled control the reproduction of the earthworms did not statistically significantly differ
- Behavioural abnormalities:
No pathological symptoms nor changes in the behaviour of the earthworms were observed in the controls or in the limit test item application rate - Results with reference substance (positive control):
- - Results with reference substance valid? Yes
- Relevant effect levels: Determined EC50 = 1.31 mg Carbendazim/kg soil DW (a.i.), which is within the range recommended by the guideline. - Reported statistics and error estimates:
- t-tests were carried out for the determination of statistically significant differences compared to the pooled control. A Normality Test and an Equal Variance Test were conducted prior to running the t-tests. P-values for both Normality and Equal Variance Test were 0.05. The a-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) was a = 0.05.
- Validity criteria fulfilled:
- yes
- Conclusions:
- Under the conditions of this study, the test item did not induce mortality, pathological symptoms or changes in the behaviour of adult earthworms after 28 days of exposure at the nominal limit test item concentration of 1000 mg/kg soil dw. The adult earthworm biomass increased in the control and at the limit test item concentration without any statistically significant differences. After eight weeks of exposure the earthworm reproduction at the limit test item concentration did not statistically significantly differ from the pooled control. As there had been no statistically significant differences determined between the control and the solvent control for the body weight and the reproduction rate, the controls were pooled.
Thus, the No Observed Effect Concentration (NOEC) for the test item summarizing the endpoints mortality, body weight and reproduction of Eisenia fetida fetida after 8 weeks of exposure in artificial soil was determined to be 1000 mg /kg soil dry weight. - Executive summary:
Effects of the test item on mortality, biomass and the reproductive potential of the earthworm species Eisenia fetida (Savigny) were determined according to the Guideline OECD 222 (2004). The study was conducted under static conditions over 8 weeks with a limit test item concentration of 1000 mg test item/kg soil dry weight, that was mixed into artificial soil containing 10 % peat. An untreated control and a solvent control were set up. 80 test organisms were inserted into 8 control, solvent control and test item replicates (10 earthworms per replicate). They had an individual body weight between 0.37 and 0.54 g at experimental starting.
After 28 days of exposure in soil, no earthworm mortalities as well as no pathological symptoms or changes in the behaviour of adult earthworms were observed in all treatments. As there were no statistically significant differences determined between the control and the solvent control for the body weight and the reproduction rate, the controls were pooled. After 28 days of exposure, no statistically significant differences in earthworm body weights at 1000 mg/kg soil dry weight were observed. After further four weeks the reproduction rate (average number of juveniles produced) was 185 in the pooled control and ranged between 112 and 244 at the limit test item concentration. Compared to the pooled control, reproduction rate was not statistically significantly reduced at 1000 mg/kg soil dry weight.
Overall, the NOEC of the test item concerning mortality, biomass and reproduction of earthworms was 1000 mg/kg soil dry weight.
All validity criteria recommended by the test guideline were fulfilled.
Summary of All Observed Effects in the Tested Application Rate
Effects
Test item
[mg test item/kg soil dry weight]
Mortality of adult earthworms
> 1000
Reduction of body weight
> 1000
Reduction of reproduction rate
(Number of juveniles)
> 1000
Summary of all effects
> 1000
Table 2: NOEC and LOEC values
Test item
Application rate
[mg test item/kg soil dry weight]
NOECreproduction, mortality, body weight
1000
LOECreproduction, mortality, body weight
> 1000
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2010-01-08 to 2010-03-09
- Reliability:
- 1 (reliable without restriction)
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
- Version / remarks:
- DIN ISO 11268-2: 1998 (E), BBA Guideline (1994)
- Deviations:
- yes
- Remarks:
- Room temperature decreased several times to a minimum of 17 °C for a total time of about 11 hours. The soil moisture of all test vessels deviated by more than 10 % of the initial moisture at the end of the test since it was decided to add additional water
- GLP compliance:
- yes
- Analytical monitoring:
- no
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Method of mixing into soil (if used): The respective test item concentration was weighed out, dissolved in acetone (2.5 mL per replicate) and mixed thoroughly with quartz sand (10 g per replicate). After complete evaporation of the solvent, the treated quartz sand was given to the artificial soil. 1164 mL demineralised water were added to adjust the humidity of the artificial soil to a moisture of 54 % of the maximum water holding capacity. Subsequently, the test medium was thoroughly mixed to ensure a homogenous distribution and about 600 g soil dry weight were filled into each test vessel.
- Controls: Artificial soil with addition of quartz sand moistened with demineralised water without test item.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone, organic solvent
- Concentration of vehicle in test medium (stock solution and final test solution): 2.5 mL per replicate
- Evaporation of vehicle before use: Yes - Test organisms (species):
- Eisenia fetida
- Animal group:
- annelids
- Details on test organisms:
- TEST ORGANISM
- Common name: Eisenia fetida SAVIGNY 1826, subspecies Eisenia fetida fetida
- Source: Own breeding
- Age at test initiation (mean and range, SD): synchronized earthworm culture, not deviating more than 1 month
- Weight at test initiation (mean and range, SD): 0.53 g
ACCLIMATION
- Acclimation period: 2 days prior to test start
- Acclimation conditions (same as test or not): same as test
- Health during acclimation (any mortality observed): healthy - Study type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- yes
- Total exposure duration:
- 56 d
- Remarks:
- Removement of adults after 28 days
- Test temperature:
- 20 +/- 2°C
- pH:
- Application rate
[mg test item/kg soil dry weight] pH-value
day 0 day 56
Control 5.75 6.25
Solvent Control 5.80 6.45
1000 5.86 6.29 - Moisture:
- Application rate
[mg test item/kg soil dry weight] Moisture [%] of dry weight
day 0 day 56
Control 37.9 43.8*
Solvent Control 37.3 44.7*
1000 38.2 42.9* - Details on test conditions:
- TEST SYSTEM
- Test container (material, size): round plastic boxes, ID: 15 cm, bottom surface area: 177 cm3, height: 14 cm, transparent and perforated lids enabled sufficient gas exchange and light penetration and prevented the test substrate from drying
- Amount of soil or substrate: 600 g DW
- No. of organisms per container (treatment): 10 (40)
- No. of replicates per treatment group: 8
- No. of replicates per control: 8
SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10 % peat, 20 % kaolin, 69 % air dried quartz sand, 0.39 % calcium carbonate
- Organic carbon (%): not determined
- Maximum water holding capacity (in % dry weigth): 73
- CEC: not determined
- Pretreatment of soil: none
- Storage (condition, duration): 18 days after preparation at test conditions in the dark
OTHER TEST CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 649 +/- 79 lx
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Adult mortality, biomass, behaviour and pathological symptoms after 28 days
Reproduction (Number of juveniles) after 56 days
VEHICLE CONTROL PERFORMED: -yes
TEST CONCENTRATIONS
- Spacing factor for test concentrations: None, limit test concentration - Nominal and measured concentrations:
- Nominal application rate: 1000 mg test substance/kg soil dry weight (DW)
- Reference substance (positive control):
- yes
- Remarks:
- 1.0 - 2.5 - 5.0 mg Carbendazim/ kg soil DW
- Duration:
- 56 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1 000 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Details on results:
- - Mortality at end of exposure period:
No noteworthy mortality of adult earthworms was observed in the controls as well as at the test item concentration of 1000 mg test item/kg soil dry weight after 28 days of exposure
- Changes in body weigth of live adults (% of initial weight) at end of exposure period:
At test start the earthworms had an individual weight of 0.30 - 0.60 g. The body weights of the earthworms increased in the control and at the limit test item concentration of 1000 mg test item/kg soil dry weight throughout the first 28 days. Compared to the control, no statistically significant difference in the increase of biomass was found
- No. of offspring produced: The mean reproduction rate was 332 in the pooled control and 375 at the limit test item concentration of 1000 mg test item/kg soil dry weight. Compared to the pooled control, the reproduction rate was not statistically significantly reduced, but increased at 1000 mg test item/kg soil dry weight. The coefficient of variation calculated for the reproduction of both controls was lower than 30 %
- Behavioural abnormalities: No symptoms or changes in the behaviour of the earthworms were observed in the controls as well as at the limit test item concentration of 1000 mg test item/kg soil dry weight after 28 days of exposure
- Other biological observations: None - Results with reference substance (positive control):
- - Results with reference substance valid? Yes
- Relevant effect levels: 2.5 mg Carbendazim/kg soil DW - Reported statistics and error estimates:
- t-tests were carried out for the determination of statistically significant differences compared to the control. A Normality Test and an Equal Variance Test were conducted prior to running the t-tests. P-values for both Normality and Equal Variance Test were 0.05. The -value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) was = 0.05. Due to a failed Normality Test reproduction data were log-transformed prior to running the ANOVA.
- Validity criteria fulfilled:
- yes
- Conclusions:
- Under the conditions of this study, Pigment Red 112 did not induce mortality or pathological symptoms of adult earthworms after an exposure to the test item concentration of 1000 mg test item/kg soil dry weight for 28 days. The biomass increase of the adult earthworms after 28 days was not statistically significantly reduced compared to the pooled control at the limit test item concentration of 1000 mg test item/kg soil dry weight. After eight weeks of exposure the earthworm reproduction was not statistically significantly reduced, but increased at the limit test item concentration of 1000 mg test item/kg soil dry weight.
Thus, the No Observed Effect Concentration (NOEC) with respect to mortality, body weight and reproduction of Eisenia fetida fetida after 8 weeks of exposure in artificial soil was determined to be at 1000 mg test item/ kg soil dry weight. - Executive summary:
Effects of Pigment Red 112 on mortality, biomass and the reproductive potential of the earthworm species Eisenia fetida (Savigny) were determined according to the guidelines DIN ISO 11268 -2 (1998), OECD 222 (2004) and BBA (1994). The study was conducted under static conditions over 8 weeks with a limit test item concentration of 1000 mg test item/kg soil dry weight, which was mixed into artificial soil containing 10 % peat. A control using untreated artificial soil and a solvent control with the solvent mixed into the artificial soil were set up. 80 test organisms, with an individual weight of 0.30 to 0.60 g, were divided into 8 replicate groups with 10 test organisms per replicate.
After 28 days no evident mortality as well as no pathological symptoms or changes in the behaviour of adult earthworms were seen in the controls as well as at the limit test item concentration of 1000 mg test item/kg soil dry weight. The biomass of the adult earthworms increased in the pooled control and at the limit test item concentration without a statistically significant difference. After further four weeks the reproduction rate (average number of juveniles) was 332 in the pooled control and 375 at the limit test item concentration. Compared to the pooled control, the reproduction rate was not statistically significantly reduced.
Overall, the NOEC of the test item concerning mortality, biomass and reproduction of earthworms was determined to be at 1000 mg test item/kg soil dry weight.
All validity criteria recommended by the test guidelines were fulfilled.
Summary of All Observed Effects in the Tested Dose
Effects
Test item
[mg test item/kg soil dry weight]
Mortality of adult earthworms
> 1000
Reduction of body weight
> 1000
Reduction of reproduction rate
(Number of juveniles)
> 1000
Summary of all effects
> 1000
NOEC and LOEC for Reproduction, Mortality and Body Weight
Test item
Application rate
[mg test item/kg soil dry weight]
NOECreproduction, mortality, body weight
1000
LOECreproduction, mortality, body weight
> 1000
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Refer to the analogue approach provided in the CSR.
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Duration:
- 56 d
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 1 000 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks on result:
- other:
- Remarks:
- Noack, 2010
- Duration:
- 56 d
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 1 000 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- reduction of body weight, mortality of adult earthworms
- Remarks on result:
- other:
- Remarks:
- Noack, 2009
- Duration:
- 56 d
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 1 000 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- mortality and reduction of body weight
- Remarks on result:
- other:
- Remarks:
- Noack, 2009b
- Duration:
- 56 d
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 1 000 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- mortaility, body weight
- Remarks on result:
- other:
- Remarks:
- Noack, 2012
Referenceopen allclose all
Mortality of Adult Earthworms in [%] after 28 Days of Exposure
Application rate [mg test item / kg soil dry weight] |
Mortality [%] |
||||
Replicate 1 |
Replicate 2 |
Replicate 3 |
Replicate 4 |
||
Control |
0 |
0 |
0 |
0 |
|
Solvent Control |
0 |
0 |
0 |
0 |
|
62.5 |
0 |
0 |
0 |
0 |
|
125 |
0 |
0 |
0 |
0 |
|
250 |
0 |
0 |
0 |
0 |
|
500 |
0 |
0 |
0 |
0 |
|
1000 |
0 |
0 |
0 |
0 |
Earthworm Behaviour and Pathological Symptoms after 28 Days of Exposure
Application rate [mg test item / kg soil dry weight] |
Effect |
Replicate |
|||
1 |
2 |
3 |
4 |
||
Control |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
Solvent Control |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
62.5 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
125 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
250 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
500 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
1000 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
Body Weight Changes of the Adult Earthworms
Application rate |
Rep. |
Mean body weights per replicate |
Mean body weight change of earthworms |
|||||
Test start |
28 days |
Per replicate |
Mean ± SD |
Sign.* |
||||
[g] |
[g] |
[g] |
[%] |
[g] |
[%] |
|||
Control |
1 |
0.481 |
0.541 |
0.06 |
12.5 |
0.10 ± 0.03 |
22.5 ± 6.42 |
- |
2 |
0.448 |
0.541 |
0.09 |
20.8 |
||||
3 |
0.439 |
0.565 |
0.13 |
28.7 |
||||
4 |
0.453 |
0.552 |
0.10 |
21.9 |
||||
Solvent Control |
1 |
0.441 |
0.570 |
0.13 |
29.3 |
|||
2 |
0.479 |
0.563 |
0.08 |
17.5 |
||||
3 |
0.447 |
0.583 |
0.14 |
30.4 |
||||
4 |
0.449 |
0.534 |
0.09 |
18.9 |
||||
62.5 |
1 |
0.479 |
0.593 |
0.11 |
23.8 |
0.13 ± 0.03 |
28.2 ± 7.74 |
No |
2 |
0.444 |
0.600 |
0.16 |
35.1 |
||||
3 |
0.451 |
0.605 |
0.15 |
34.2 |
||||
4 |
0.452 |
0.540 |
0.09 |
19.5 |
||||
125 |
1 |
0.451 |
0.580 |
0.13 |
28.6 |
0.13 ± 0.01 |
28.2 ± 3.63 |
No |
2 |
0.451 |
0.565 |
0.11 |
25.3 |
||||
3 |
0.478 |
0.601 |
0.12 |
25.7 |
||||
4 |
0.437 |
0.582 |
0.15 |
33.2 |
||||
250 |
1 |
0.477 |
0.575 |
0.10 |
20.5 |
0.12 ± 0.03 |
27.1 ± 6.57 |
No |
2 |
0.448 |
0.594 |
0.15 |
32.6 |
||||
3 |
0.453 |
0.554 |
0.10 |
22.3 |
||||
4 |
0.444 |
0.590 |
0.15 |
32.9 |
||||
500 |
1 |
0.444 |
0.581 |
0.14 |
30.9 |
0.12 ± 0.03 |
27.3 ± 7.81 |
No |
2 |
0.442 |
0.581 |
0.14 |
31.4 |
||||
3 |
0.454 |
0.596 |
0.14 |
31.3 |
||||
4 |
0.475 |
0.549 |
0.07 |
15.6 |
||||
1000 |
1 |
0.454 |
0.599 |
0.15 |
31.9 |
0.14 ± 0.02 |
29.9 ± 5.07 |
No |
2 |
0.445 |
0.606 |
0.16 |
36.2 |
||||
3 |
0.480 |
0.606 |
0.13 |
26.3 |
||||
4 |
0.441 |
0.553 |
0.11 |
25.4 |
Reproduction Rate (Number of Juveniles after 8 Weeks)
Application rate |
Rep. |
Number of juveniles |
Mean ± SD |
CV [%] |
[%] of pooled control |
Significance compared to pooled control |
Control |
1 |
359 |
359 ± 49.6 |
13.8 |
- |
- |
2 |
338 |
|||||
3 |
456 |
|||||
4 |
323 |
|||||
Solvent Control |
1 |
354 |
||||
2 |
388 |
|||||
3 |
287 |
|||||
4 |
365 |
|||||
62.5 |
1 |
338 |
365 ± 28.5 |
7.81 |
102 |
No |
2 |
356 |
|||||
3 |
360 |
|||||
4 |
405 |
|||||
125 |
1 |
284 |
355 ± 60.2 |
17.0 |
98.9 |
No |
2 |
327 |
|||||
3 |
393 |
|||||
4 |
415 |
|||||
250 |
1 |
378 |
347 ± 88.3 |
25.5 |
96.6 |
No |
2 |
442 |
|||||
3 |
334 |
|||||
4 |
232 |
|||||
500 |
1 |
303 |
288 ± 49.1 |
17.1 |
80.3 |
No |
2 |
250 |
|||||
3 |
351 |
|||||
4 |
248 |
|||||
1000 |
1 |
344 |
346 ± 83.8 |
24.2 |
96.4 |
No |
2 |
385 |
|||||
3 |
424 |
|||||
4 |
230 |
Mortality of Adult Earthworms in [%] after 28 Days of Exposure
Application rate [mg test item/kg soil dry weight] |
Mortality [%] |
||||
Replicate 1 |
Replicate 2 |
Replicate 3 |
Replicate 4 |
||
Control |
0 |
0 |
0 |
0 |
|
Solvent Control |
0 |
0 |
0 |
0 |
|
62.5 |
0 |
0 |
0 |
0 |
|
125 |
0 |
0 |
0 |
0 |
|
250 |
0 |
0 |
0 |
0 |
|
500 |
0 |
0 |
0 |
0 |
|
1000 |
0 |
0 |
0 |
0 |
Earthworm Behaviour and Pathological Symptoms after 28 Days of Exposure
Application rate [mg test item/kg soil dry weight] |
Effect |
Replicate |
|||
1 |
2 |
3 |
4 |
||
Control |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
Solvent Control |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
62.5 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
125 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
250 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
500 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
1000 |
a) |
10/10 |
10/10 |
10/10 |
10/10 |
a) No observation of symptoms or behavioural changes
Body Weight Changes of the Adult Earthworms
Application rate |
Rep. |
Mean body weights per replicate |
Mean body weight change of earthworms |
|||||
Test start |
28 days |
Per replicate |
Mean ± SD |
Sign.* |
||||
[g] |
[g] |
[g] |
[%] |
[g] |
[%] |
|||
Control |
1 |
0.498 |
0.723 |
0.23 |
45.2 |
0.23 ± 0.04 |
46.8 ± 7.47 |
- |
2 |
0.520 |
0.786 |
0.27 |
51.2 |
||||
3 |
0.498 |
0.672 |
0.17 |
34.9 |
||||
4 |
0.500 |
0.720 |
0.22 |
44.0 |
||||
Solvent Control |
1 |
0.474 |
0.768 |
0.29 |
62.0 |
|||
2 |
0.471 |
0.685 |
0.21 |
45.4 |
||||
3 |
0.496 |
0.716 |
0.22 |
44.4 |
||||
4 |
0.502 |
0.740 |
0.24 |
47.4 |
||||
62.5 |
1 |
0.498 |
0.720 |
0.22 |
44.6 |
0.23 ± 0.01 |
46.0 ± 2.09 |
No |
2 |
0.503 |
0.744 |
0.24 |
47.9 |
||||
3 |
0.499 |
0.718 |
0.22 |
43.9 |
||||
4 |
0.503 |
0.743 |
0.24 |
47.7 |
||||
125 |
1 |
0.465 |
0.668 |
0.20 |
43.7 |
0.25 ± 0.05 |
50.7 ± 11.3 |
No |
2 |
0.473 |
0.791 |
0.32 |
67.2 |
||||
3 |
0.500 |
0.744 |
0.24 |
48.8 |
||||
4 |
0.499 |
0.715 |
0.22 |
43.3 |
||||
250 |
1 |
0.471 |
0.703 |
0.23 |
49.3 |
0.28 ± 0.04 |
57.1 ± 8.37 |
No |
2 |
0.467 |
0.789 |
0.32 |
69.0 |
||||
3 |
0.499 |
0.776 |
0.28 |
55.5 |
||||
4 |
0.503 |
0.778 |
0.28 |
54.7 |
||||
500 |
1 |
0.466 |
0.754 |
0.29 |
61.8 |
0.27 ± 0.02 |
56.4 ± 4.73 |
No |
2 |
0.470 |
0.739 |
0.27 |
57.2 |
||||
3 |
0.502 |
0.784 |
0.28 |
56.2 |
||||
4 |
0.501 |
0.753 |
0.25 |
50.3 |
||||
1000 |
1 |
0.469 |
0.727 |
0.26 |
55.0 |
0.28 ± 0.02 |
57.9 ± 2.50 |
No |
2 |
0.470 |
0.736 |
0.27 |
56.6 |
||||
3 |
0.501 |
0.804 |
0.30 |
60.5 |
||||
4 |
0.504 |
0.803 |
0.30 |
59.3 |
Rep. = Replicate; SD = Standard deviation; Sign. = Significance; * Significance compared to pooled control
Reproduction Rate (Number of Juveniles after 8 Weeks)
Application rate |
Rep. |
Number of juveniles |
Mean ± SD |
CV [%] |
[%] of pooled control |
Significance compared to pooled control |
Control |
1 |
227 |
235 ± 39.1 |
16.7 |
- |
- |
2 |
184 |
|||||
3 |
203 |
|||||
4 |
275 |
|||||
Solvent Control |
1 |
219 |
||||
2 |
211 |
|||||
3 |
260 |
|||||
4 |
298 |
|||||
62.5 |
1 |
279 |
244 ± 30.4 |
12.5 |
104 |
No |
2 |
212 |
|||||
3 |
258 |
|||||
4 |
226 |
|||||
125 |
1 |
284 |
238 ± 61.6 |
25.8 |
102 |
No |
2 |
165 |
|||||
3 |
294 |
|||||
4 |
210 |
|||||
250 |
1 |
238 |
269 ± 42.6 |
15.8 |
115 |
No |
2 |
231 |
|||||
3 |
284 |
|||||
4 |
322 |
|||||
500 |
1 |
182 |
237 ± 67.5 |
28.5 |
101 |
No |
2 |
335 |
|||||
3 |
209 |
|||||
4 |
221 |
|||||
1000 |
1 |
299 |
267 ± 31.4 |
11.8 |
114 |
No |
2 |
288 |
|||||
3 |
236 |
|||||
4 |
244 |
SD = Standard deviation CV = Coefficient of variation Rep. = Replicate
Mortality of Adult Earthworms in [%] after 28 Days of Exposure
Application rate [mg test item/kg soil dry weight] |
Mortality [%] |
||||||||
Replicate |
|||||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
Mean |
|
Control |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Solvent Control |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1000 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Earthworm Behaviour and Pathological Symptoms after 28 Days of Exposure
Application rate [mg test item/kg soil dry weight] |
Effect |
Replicate |
|||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
||
Control |
A) |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
Solvent Control |
A) |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
1000 |
A) |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
A) No observation of symptoms or behavioural changes.
Body Weight Changes of the Adult Earthworms
Application rates [mg test item /kg soil dry weight] |
Replicate |
Mean body weights per replicate |
Mean body weight change of earthworms |
|||||
Test start |
28 days |
Per replicate |
Mean ± SD |
Sig. to pooled control |
||||
[g] |
[g] |
[g] |
[%] |
[g] |
[%] |
|||
Control |
1 |
0.43 |
0.66 |
0.23 |
53.5 |
0.19 ± 0.03 |
43.5 ± 7.91 |
- |
2 |
0.44 |
0.60 |
0.16 |
36.4 |
||||
3 |
0.42 |
0.65 |
0.23 |
54.8 |
||||
4 |
0.45 |
0.64 |
0.19 |
42.2 |
||||
5 |
0.43 |
0.64 |
0.21 |
48.8 |
||||
6 |
0.44 |
0.59 |
0.15 |
34.1 |
||||
7 |
0.42 |
0.59 |
0.17 |
40.5 |
||||
8 |
0.45 |
0.62 |
0.17 |
37.8 |
||||
Solvent Control |
1 |
0.45 |
0.63 |
0.18 |
40.0 |
0.17 ± 0.03 |
39.1 ± 5.98 |
- |
2 |
0.43 |
0.64 |
0.21 |
48.8 |
||||
3 |
0.44 |
0.58 |
0.14 |
31.8 |
||||
4 |
0.42 |
0.58 |
0.16 |
38.1 |
||||
5 |
0.45 |
0.66 |
0.21 |
46.7 |
||||
6 |
0.43 |
0.58 |
0.15 |
34.9 |
||||
7 |
0.44 |
0.59 |
0.15 |
34.1 |
||||
8 |
0.42 |
0.58 |
0.16 |
38.1 |
||||
Pooled Control |
0.18 ± 0.03 |
41.3 ± 7.15 |
- |
|||||
1000 |
1 |
0.42 |
0.60 |
0.18 |
42.9 |
0.19 ± 0.03 |
44.6 ± 7.59 |
No |
2 |
0.45 |
0.64 |
0.19 |
42.2 |
||||
3 |
0.43 |
0.67 |
0.24 |
55.8 |
||||
4 |
0.44 |
0.60 |
0.16 |
36.4 |
||||
5 |
0.42 |
0.66 |
0.24 |
57.1 |
||||
6 |
0.45 |
0.64 |
0.19 |
42.2 |
||||
7 |
0.43 |
0.60 |
0.17 |
39.5 |
||||
8 |
0.44 |
0.62 |
0.18 |
40.9 |
Reproduction Rate (Number of Juveniles after 8 Weeks)
Application rate [mg test item/kg soil dry weight] |
Rep. |
Number of juveniles |
Mean ± SD |
CV [%] |
[%] of pooled control |
Significance compared to pooled control |
Control |
1 |
149 |
173 ± 41.9 |
24.2 |
– |
– |
2 |
259 |
|||||
3 |
131 |
|||||
4 |
199 |
|||||
5 |
156 |
|||||
6 |
151 |
|||||
7 |
193 |
|||||
8 |
146 |
|||||
Solvent Control |
1 |
177 |
197 ± 36.8 |
18.7 |
– |
– |
2 |
140 |
|||||
3 |
168 |
|||||
4 |
233 |
|||||
5 |
230 |
|||||
6 |
172 |
|||||
7 |
231 |
|||||
8 |
226 |
|||||
Pooled Control |
185 ± 40.1 |
21.7 |
– |
– |
||
1000 |
1 |
244 |
164 ± 51.5 |
31.4 |
88.6 |
No |
2 |
236 |
|||||
3 |
112 |
|||||
4 |
144 |
|||||
5 |
170 |
|||||
6 |
167 |
|||||
7 |
129 |
|||||
8 |
113 |
Mortality of Adult Earthworms in [%] after 28 Days of Exposure
Application rate [mg test item/kg soil dry weight] |
Mortality [%] |
||||||||
Replicate |
|||||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
Mean |
|
Control |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Solvent Control |
0 |
0 |
0 |
10 |
0 |
0 |
0 |
0 |
1.25 |
1000 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Earthworm Behaviour and Pathological Symptoms after 28 Days of Exposure
Application rate [mg test item/kg soil dry weight] |
Effect |
Replicate |
|||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
||
Control |
A) |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
Solvent Control |
A) |
10/10 |
10/10 |
10/10 |
9/10 |
10/10 |
10/10 |
10/10 |
10/10 |
H) |
– |
– |
– |
1/10 |
– |
– |
– |
– |
|
1000 |
A) |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
10/10 |
Body Weight Changes of the Adult Earthworms
Application rate kg soil dry weight] |
Rep. |
Mean body weights per replicate |
Mean body weight change of earthworms |
||||
Test start |
28 days |
Per replicate |
Mean ± SD [%] |
Significance compared to pooled control with [%] data |
|||
[g] |
[g] |
[g] |
[%] |
||||
Control |
1 |
0.573 |
0.827 |
0.25 |
44.3 |
54.3 |
– |
2 |
0.548 |
0.805 |
0.26 |
46.9 |
|||
3 |
0.498 |
0.800 |
0.30 |
60.6 |
|||
4 |
0.526 |
0.830 |
0.30 |
57.8 |
|||
5 |
0.535 |
0.799 |
0.26 |
49.3 |
|||
6 |
0.494 |
0.819 |
0.33 |
65.8 |
|||
7 |
0.516 |
0.781 |
0.27 |
51.4 |
|||
8 |
0.520 |
0.822 |
0.30 |
58.1 |
|||
Solvent Control |
1 |
0.524 |
0.817 |
0.29 |
55.9 |
53.4 |
– |
2 |
0.555 |
0.805 |
0.25 |
45.0 |
|||
3 |
0.553 |
0.794 |
0.24 |
43.6 |
|||
4 |
0.504 |
0.869 |
0.36 |
72.4 |
|||
5 |
0.529 |
0.811 |
0.28 |
53.3 |
|||
6 |
0.535 |
0.788 |
0.25 |
47.3 |
|||
7 |
0.486 |
0.773 |
0.29 |
59.1 |
|||
8 |
0.519 |
0.780 |
0.26 |
50.3 |
|||
1000 |
1 |
0.518 |
0.790 |
0.27 |
52.5 |
48.0 |
No |
2 |
0.519 |
0.763 |
0.24 |
47.0 |
|||
3 |
0.565 |
0.831 |
0.27 |
47.1 |
|||
4 |
0.545 |
0.792 |
0.25 |
45.3 |
|||
5 |
0.495 |
0.715 |
0.22 |
44.4 |
|||
6 |
0.525 |
0.779 |
0.25 |
48.4 |
|||
7 |
0.525 |
0.743 |
0.22 |
41.5 |
|||
8 |
0.490 |
0.774 |
0.28 |
58.0 |
Reproduction Rate (Number of Juveniles after 8 Weeks)
Application rate [mg test item/kg soil dry weight] |
Rep. |
Number of juveniles |
Mean ± SD |
CV [%] |
[%] of control |
Significance compared to pooled control |
Control |
1 |
340 |
333 ± 35.7 |
10.7 |
– |
– |
2 |
299 |
|||||
3 |
356 |
|||||
4 |
288 |
|||||
5 |
378 |
|||||
6 |
290 |
|||||
7 |
365 |
|||||
8 |
350 |
|||||
Solvent Control |
1 |
328 |
332 ± 56.3 |
17.0 |
– |
– |
2 |
293 |
|||||
3 |
290 |
|||||
4 |
272 |
|||||
5 |
339 |
|||||
6 |
455 |
|||||
7 |
340 |
|||||
8 |
335 |
|||||
1000 |
1 |
337 |
375 ± 25.0 |
6.68 |
113 |
Yes* |
2 |
354 |
|||||
3 |
362 |
|||||
4 |
395 |
|||||
5 |
361 |
|||||
6 |
410 |
|||||
7 |
390 |
|||||
8 |
392 |
Description of key information
EC10 / NOEC soil macroorganisms ≥ 1000 mg/kg soil dw
Key value for chemical safety assessment
Additional information
No toxicity of Pigments Orange 38, Red 2, Red 112 and Red 187 to the earthworm species Eisenia fetida was found in reliable (category 1) Earthworm Reproduction Tests over a period of 8 weeks (OECD 222, 2004). Either tests with a nominal limit concentration of 1000 mg/kg soil dry weight and 80 test organisms divided into 8 replicate groups were performed (Pigment Red 2 and 112), or five test item concentrations from 62.5 to 1000 mg/kg were tested with 40 test organisms per treatment group divided into 4 replicate groups. None of the four tested pigments did induce mortality or pathological symptoms to adult earthworms and the biomass increase of the adult earthworms was not statistically significantly reduced compared to the control after 28 days of exposure. In none of the four tests any statistically significantly reduced reproduction of earthworms was observed compared to the control. All studies were performed compliant to GLP meeting the validity criteria of the OECD guideline. Therefore, Naphtol-AS Pigments are to be regarded as chronically non-toxic to soil macroorganisms.
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.