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EC number: 205-426-2 | CAS number: 140-66-9
- 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, other
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- In accordance with Regulation (EC) 1907/2006 Annex XI (1.5) and the relevant ECHA guidance documents, the substances detailed in the table below are grouped for the purposes of read across to reduce the need for unnecessary repeat testing on the basis that the substances are similar on the basis of a common functional groups.
- 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:
- 4 wk
- Dose descriptor:
- EC10
- Effect conc.:
- 24 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks on result:
- other: (5.2, 110.7)
- Duration:
- 8 wk
- Dose descriptor:
- EC10
- Effect conc.:
- 55.8 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Duration:
- 8 wk
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Mean number of live juvenile earthworms (offspring)
- Duration:
- 14 d
- Dose descriptor:
- LC50
- Effect conc.:
- 88.6 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Conclusions:
- The read across for 4-tert-octylphenol (CAS: 140-66-9); is based upon the analogous substances to which basic form, degree of substitution of functional groups is not considered to effect the proposed read across for the endpoint of Toxicity to soil macroorganisms except arthropods. Based on the information available for the read across substances, the substance is expected to be toxic to soil macroorganisms except arthropods .
- Endpoint:
- toxicity to soil macroorganisms except arthropods: short-term
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 2001-2004
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- The document is a report to the UK Government and differs in style and content from a laboratory study report and a peer-reviewed publication. However, it contains full details on the methods used in the studies that support the paper, and its only issue appears to be that it does not state whether these are conducted to GLP. Therefore, it has been assigned a KIlimisch score of 2.
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- according to guideline
- Guideline:
- ISO 11268-1 (Effects of Pollutants on Earthworms. 1. Determination of Acute Toxicity Using Artificial Soil Substrate)
- Deviations:
- yes
- Remarks:
- Slight deviation of standard temperature but this is not expected to affect the results.
- GLP compliance:
- no
- Specific details on test material used for the study:
- Details on properties of test surrogate or analogue material (migrated information):
No data - Analytical monitoring:
- no
- Details on sampling:
- - Start of Test: Samples of soil were collected from each vessel at commencement of testing and stored at -20 °C prior to analysis.
- End of Test: Live worms were snap frozen in liquid nitrogen, then stored at -80 °C prior to analysis. - Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Vehicle: Acetone
- Preparation: For each exposure concentration a stock solution of the test substance was prepared in acetone and added to a quantity of soil such that the maximum volume of acetone used was 3.2 mg/kg. After spiking the soil, 0.5 kg was added to replicate test vessels and then left overnight to allow any remaining acetone to volatilise before adding the test animals.
- Controls: Negative control and solvent (acetone) controls were included in the study design. - Test organisms (species):
- Eisenia sp.
- Animal group:
- annelids
- Details on test organisms:
- - Supplier: Earthworms were purchased from a commercial supplier (details not available).
- Study type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- no
- Total exposure duration:
- 14 d
- Post exposure observation period:
- Surviving earthworms were retained for biomarker analysis in relation to endocrine disruption studies.
- Test temperature:
- 25 ± 2 °C
- pH:
- 6.0 ± 0.5 (in accordance with the OECD Artificial Soil)
- Moisture:
- 25 to 42% of the dry weight soil (in accordance with the OECD Artificial Soil)
- Details on test conditions:
- - Exposure: After spiking the soils, 0.5 kg of soil was added to each test vessel and left overnight for the acetone to volatilise before the earthworms were added to the test vessels.
- Test vessel: Kilner jars (glass).
- Number of organisms per concentration at the start of the test: 20 in control and solvent control; 10 in all test substance concentrations.
- Acclimation: Earthworms were added to each replicate and allowed to burrow into the soil.
- Food: 5 g of dried ground rabbit's droppings were placed on the surface of the soil in each vessel for the worms to feed on.
- Photoperiod: 16 hours light and 8 hours dark cycle.
- Weight of vessels: The weights of the vessels were recorded twice weekly so that weights could be adjusted to compensate for evaporative losses by the addition of distilled water.
- Removal: On day 14, live and dead adult earthworms were removed from the test vessels by sieving. - Nominal and measured concentrations:
- Nominal Concentrations: 1.0, 3.2, 10, 32, 100, 320, 1000, 3200, 10000 mg/kg.
- Reference substance (positive control):
- no
- Duration:
- 14 d
- Dose descriptor:
- LC50
- Effect conc.:
- 88.6 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Details on results:
- - Quality Control: QC criteria were met with 0% lethality in the negative control and 10% lethality in the solvent control.
- Results with reference substance (positive control):
- No data
- Reported statistics and error estimates:
- Statistics were used but the method is not reported. ENVIRON UK Ltd has statistically derived an LC50 using Probit analysis assuming up to 3 mortalities per control (i.e., less than 10% mortality). The statistical tool used was ProStat Ver. 5.01 for Windows by Poly Software International, Inc., P.O. Box 60, Pearl River, NY 10965 USA. The wide 95% confidence intervals seemed to be a function of control mortality.
- Validity criteria fulfilled:
- yes
- Conclusions:
- Nonylphenol was found to affect the survival of adult earthworms (Eisenia sp.) with a NOEC of 32 mg/kg and a LOEC of 100 mg/kg dry weight soil. This study was a precursor to further studies on the long-term effects of nonylphenol and other endocrine disrupting substances to earthworms and the aquatic amphipod Gammarus pulex. Further endpoints studied were reproduction and three genetic biomarkers (vitellogenin, a sperm marker and gene expression of annetocin). It was concluded that although earthworms are important test species for assessing the effects of substances to terrestrial ecosystems, the hermaphroditic nature of earthworms means that they are not appropriate for the assessment of endocrine disrupting effects.
- Executive summary:
In a 14-day acute toxicity study, earthworms (Eisenia andreii) were exposed at control, solvent control, and nonylphenol at 1.0, 3.2, 10, 32, 100, 320, 1000, 3200, 10000 mg nonylphenol/kg artificial soil. The NOEC based on lethality was 32 mg nonylphenol/kg, with a LOEC of 100 mg nonylphenol/kg, respectively. ENVIRON derived a statistical LC50 of 88.6 mg nonylphenol/kg which is taken as the key value for assessment. Based on the results of this study, nonylphenol would be considered to have adverse acute effects to Eisenia andreii.
This study is classified as acceptable and is similar in methodology to the requirements of the ISO Guideline 11268 -2; Effects of Pollutants on Earthworms (Part 2, Determination of Effects on Reproduction).
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2001-2004
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- The document is a report to the UK Government and differs in style and content from a laboratory study report and a peer-reviewed publication. However, it contains full details on the methods used in the studies that support the paper, and its only issue appears to be that it does not state whether these are conducted to GLP. Therefore, it has been assigned a KIlimisch score of 2.
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- according to guideline
- Guideline:
- ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)
- Deviations:
- yes
- Remarks:
- Slight deviation of standard temperature but this is not expected to affect the results.
- GLP compliance:
- no
- Specific details on test material used for the study:
- Details on properties of test surrogate or analogue material (migrated information):
No data - Analytical monitoring:
- no
- Details on sampling:
- - Start of Test: Samples of soil were collected from each vessel at commencement of testing and stored at -20 °C prior to analysis.
- End of Test: Live worms were snap frozen in liquid nitrogen, then stored at -80 °C prior to analysis. - Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Vehicle: Acetone
- Preparation: For each exposure concentration a stock solution of the test substance was prepared in acetone and added to a quantity of soil such that the maximum volume of acetone used was 3.2 mg/kg. After spiking the soil, 0.5 kg was added to replicate test vessels and then left overnight to allow any remaining acetone to volatilise before adding the test animals.
- Controls: Negative control and solvent (acetone) controls were included in the study design. - Test organisms (species):
- Eisenia sp.
- Animal group:
- annelids
- Details on test organisms:
- - Supplier: Earthworms were purchased from a commercial supplier (details not available).
- Study type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- no
- Total exposure duration:
- 8 wk
- Post exposure observation period:
- Surviving earthworms were retained for biomarker analysis in relation to endocrine disruption studies.
- Test temperature:
- 25 ± 2 °C
- pH:
- 6.0 ± 0.5 (in accordance with the OECD Artificial Soil)
- Moisture:
- 25 to 42% of the dry weight soil (in accordance with the OECD Artificial Soil)
- Details on test conditions:
- - Exposure: After spiking the soils, 0.5 kg of soil was added to each test vessel and left overnight for the acetone to volatilise before the earthworms were added to the test vessels.
- Test vessel: Kilner jars (glass).
- Number of organisms per concentration at the start of the test: 40 in control and solvent control; 40 in all test substance concentrations.
- Acclimation: Earthworms were added to each replicate and allowed to burrow into the soil.
- Food: 5g of dried ground rabbit's droppings were placed on the surface of the soil in each vessel for the worms to feed on.
- Photoperiod: 16 hours light and 8 hours dark cycle.
- Weight of vessels: The weights of the vessels were recorded twice weekly so that weights could be adjusted to compensate for evaporative losses by the addition of distilled water.
- Removal: On day 28, adult worms were removed from the test vessels by sieving then snap frozen in liquid nitrogen for later analysis. Sieved soil containing cocoons was returned to the vessels and left for a further 28 days to allow cocoons to hatch. At the end of the test (56 days) the number of juvenile worms in each vessel was counted after wet-sieving. - Nominal and measured concentrations:
- Nominal Concentrations: 1.0, 3.2, 10, 32, 100 mg/kg.
- Reference substance (positive control):
- no
- Duration:
- 8 wk
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Mean number of live juvenile earthworms (offspring)
- Details on results:
- - Quality Control: QC criteria were met. Mean number of live juvenile worms at day 56 in negative control was 27.6 ± 14.6 (standard deviation) and in the solvent control was 28.9 ± 20.7. The mean number of live juvenile worms at day 56 in the highest concentration tested (100 mg/kg) was 23.0 ± 10.0 (standard deviation).
- Results with reference substance (positive control):
- No data
- Reported statistics and error estimates:
- Statistics were used but the method is not reported.
- Validity criteria fulfilled:
- yes
- Conclusions:
- Nonylphenol was not found to affect the reproduction of adult earthworms (Eisenia sp.) with a NOEC of 100 mg/kg dry weight soil. This study was part of a broader study of the long-term effects of nonylphenol and other endocrine disrupting substances to earthworms and the aquatic amphipod Gammarus pulex. Further endpoints studied included three genetic biomarkers (vitellogenin, a sperm marker and gene expression of of annetocin). It was concluded that although earthworms are important test species for assessing the effects of substances to terrestrial ecosystems, the hermaphroditic nature of earthworms means that they are not appropriate for the assessment of endocrine disrupting effects.
- Executive summary:
In an eight week chronic toxicity study, earthworms (Eisenia andreii) were exposed at control, solvent control, and nonylphenol at 1.0, 3.2, 10, 32, 100 mg nonylphenol/kg artificial soil. The NOEC based on reproduction was 100 mg nonylphenol/kg. Based on the results of this study, nonylphenol would not be considered to have adverse reproductive effects to Eisenia andreii.
This study is classified as acceptable and is similar in methodology to the requirements of the ISO Guideline 11268 -2; Effects of Pollutants on Earthworms (Part 2, Determination of Effects on Reproduction).
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- Spring 2008
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Published paper which meets international standard methods including OECD and ISO Guidelines.
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- according to guideline
- Guideline:
- other: ISO Guideline 16387 Soil Quality - effects of pollutants on Enchytraeidae (Enchytraeus sp.) - determination of effects on survival and reproduction. Guideline No. 16387. International Organisation for Standardisation, Geneva, pp 1-22 (ISO 2003)
- Deviations:
- yes
- Remarks:
- The test vessels and volumes differed from the ISO protocol but this is not thought to have affected the results of the experiment. In addition, the test period was shortened in accordance with Kuperman et al (2006) as E. crypticus has a shorter generatio
- GLP compliance:
- no
- Specific details on test material used for the study:
- Details on properties of test surrogate or analogue material (migrated information):
No data - Analytical monitoring:
- no
- Details on sampling:
- No data
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Vehicle: Nonylphenol was applied to soils dissolved in acetone (95%, Panreac, Barcelona, Spain).
- Application: To prepare the concentrations of test substance, the amount of soil to be used for each concentration was moistened with a fixed volume of acetone solution containing the appropriate quantity of nonylphenol. Acetone was then left to evaporate for 24 hours in a fume hood.
- Controls: Two control soils were prepared; one negative control and one solvent control to which the same volume of acetone was added as was used in the test substance concentrations.
- Concentrations: The test concentrations were selected following a preliminary assay consisting of 0, 1, 10, 100, 1000 and 10000 mg of nonylphenol per kg of soil. The range of concentrations showing an inhibition between 10% and 90% were selected for the study. - Test organisms (species):
- other: Enchytraeus crypticus
- Animal group:
- annelids
- Details on test organisms:
- No further data on test organisms is provided.
- Study type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- yes
- Total exposure duration:
- 4 wk
- Remarks:
- This is a shorter exposure period than the standard 6-week duration in the ISO Protocol. However, the modifications were made because the enchytraeid species is more easily damaged and has a shorter generation time than E. albidus.
- Post exposure observation period:
- No data
- Test temperature:
- 21 °C
- pH:
- 6 ± 0.5 (as per OECD Artificial Soil) adjusted using calcium carbonate.
- Moisture:
- Water holding capacity: 44.8%
Moisture content: Soil water content was adjusted to 60% of the maximum water holding capacity. - Details on test conditions:
- TEST SYSTEM
- OECD artificial soil: Prepared according to the OECD protocol for artificial soil (1984) by mixing Sphagnum peat (10%), kaoline (20%) and quartz sand.
- Testing facility: Universitat Autonoma de Barcelona
- Test container (type, material, size): Each replicate consisted of a 150-ml polyethylene flask.
- Amount of soil: Each vessel was filled with 30 g (dry weight)
- No. of replicates per treatment group: 5 replicates per concentration were prepared
- Number of organisms: Ten adults (clearly identified by the clitella were introduced into each flask.
- Feeding: Animals were fed with 25 mg of ground oats at the start and weekly thereafter.
- Photoperiod: Test was maintained in the dark.
- Aeration: Animals were aerated twice a week.
SOURCE AND PROPERTIES OF SUBSTRATE
- Geographic location: The results presented here relate to the OECD Artificial Soil, however two natural soils were also tested. These correspond to a loamy agricultural soil (PRA) within the Universitat Autonoma de Barcelona campus (Cerdanyola del Valles, NE Spain) and the other was a loam sand soil (UAB) from the dry grassland in Serra de Prades (Tarragona, NE Spain).
- Pesticide use history at the collection site: Not relevant to the OECD Artificial Soil, however, pesticide use had ceased five years previously in the agricultural soil (UAB).
- Collection procedures: Natural soils were defaunated by alternating two consecutive freeze-thaw cycles (-20 °C for 4 days, then +20 °C for 4 days).
- Sampling depth (cm): Not relevant to the OECD soil, but natural soils were collected from the upper layer (20 cm).
- Soil texture (if natural soil): PRA soil
- % sand: 80.3
- % silt: 16.5
- % clay: 3.2
- Soil texture (if natural soil): UAB soil
- % sand: 36.4
- % silt: 44.9
- % clay: 18.7
- Composition (if artificial substrate): OECD Artificial Soil
- Organic carbon (%): 3.45
- Maximum water holding capacity (in % dry weight): 44.8% adjusted to 60% WHC
- CEC: 8.94
END OF TEST
- Adults and juveniles were fixed by direct addition of ethanol (80% v/v) to each flask.
- A few drops of Bengal red (1% solution in ethanol) were added to colour the individuals. After 2 hours, individuals were separated from the more fine soil particles by rinsing soil with tap water in a sieve (0.2 mm) and then transferred to a Petri dish for counting.
- All the individuals counted were assumed to be alive given the quick degradation time of individuals once dead.
METHOD VARIATION
- The test was shortened from 6 weeks to 4 weeks in accoradance with Kuperman et al (2006). The full reference is Kuperman RG, Checkai RT, Simini M, Phillips CT, Anthony JS, Kolakowski JE and Davis EA (2006) Toxicity of emerging energetic soil contaminant CL-20 to potworm Enchytraeus crypticus in freshly amended or weathered and aged treatments. Chemosphere 62: 1282-1293. - Nominal and measured concentrations:
- Nominal concentrations: The test concentrations were selected following a preliminary assay consisting of 0, 1, 10, 100, 1000 and 10000 mg of nonylphenol per kg of soil. The range of concentrations showing an inhibition between 10% and 90% were selected for the study.
- Reference substance (positive control):
- no
- Duration:
- 4 wk
- Dose descriptor:
- EC10
- Effect conc.:
- 24 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks on result:
- other: (5.2, 110.7)
- Details on results:
- No differences in the outcomes between controls with and without the addition of acetone were found in any of the assays. Validity criteria were met for all species.
- Survival was over 80%
- Reproduction was over 25 juveiles with a coefficient of variation below 50%.
Mean values (+/- standard deviation) are provided for the controls in the different test soils below:
- Survival in adults: OECD 9.0 (±1.8); PRA 7.8 (±3.0); UAB 8.8 (±0.9)
- Reproduction in juveniles: OECD 391.8 (±179.6); PRA 702.0 (±322.8); UAB 777.2 (±311.3) - Results with reference substance (positive control):
- No data
- Reported statistics and error estimates:
- - EC10 was calculated using Statistica 6.0 (StatSoft Inc., Tulsa USA).
- The influence of soil properties was assessed by Generalised Linear Models (Brodgar 2.0, Highland Statistics Ltd, Newburgh, UK). - Validity criteria fulfilled:
- yes
- Conclusions:
- The authors considered nonylphenol to be more toxic than the nonylphenol ethoxylate mixture and considered the results to be in agreement with existing published literature. Nonylphenol presented low phytotoxicity but moderate toxicity for invertebrates, reproduction being clearly the most sensitive endpoint. Nonylphenol generally affected the invertebrates' reproduction at lower concentrations than those affecting growth or survival. Earthworms and collembolans were more sensitive taxa compared to enchytraeids.
- Executive summary:
In a 4-week chronic toxicity study, enchytraeids (Enchytraeus crypticus) were exposed at control, solvent control, and nonylphenol at a range of concentrations showing inhibition between 10% and 90% from an initial range-finding test. The EC10 based on reproduction was 24 mg nonylphenol/kg, using an artificial soil test medium. Results for two natural soils were also reported. Based on the results of this study, nonylphenol would be considered to have adverse chronic and acute effects to Enchytraeus crypticus.
This study is classified as acceptable and is similar in methodology to the requirements of the ISO Guideline 16387 (ISO, 2003).
- Endpoint:
- toxicity to soil macroorganisms except arthropods: long-term
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- Spring 2008
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Published paper which meets international standard methods including OECD and ISO Guidelines.
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- according to guideline
- Guideline:
- ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)
- Deviations:
- no
- GLP compliance:
- no
- Specific details on test material used for the study:
- Details on properties of test surrogate or analogue material (migrated information):
No data - Analytical monitoring:
- no
- Details on sampling:
- No data
- Vehicle:
- yes
- Details on preparation and application of test substrate:
- - Vehicle: Nonylphenol was applied to soils dissolved in acetone (95%, Panreac, Barcelona, Spain).
- Application: To prepare the concentrations of test substance, the amount of soil to be used for each concentration was moistened with a fixed volume of acetone solution containing the appropriate quantity of nonylphenol. Acetone was then left to evaporate for 24 hours in a fume hood.
- Controls: Two control soils were prepared; one negative control and one solvent control to which the same volume of acetone was added as was used in the test substance concentrations.
- Concentrations: The test concentrations were selected following a preliminary assay consisting of 0, 1, 10, 100, 1000 and 10000 mg of nonylphenol per kg of soil. The range of concentrations showing an inhibition between 10% and 90% were selected for the study. - Test organisms (species):
- Eisenia sp.
- Animal group:
- annelids
- Details on test organisms:
- No further data on test organisms is provided.
- Study type:
- laboratory study
- Substrate type:
- artificial soil
- Limit test:
- yes
- Total exposure duration:
- 8 wk
- Post exposure observation period:
- No data
- Test temperature:
- 21 °C
- pH:
- 6 ± 0.5 (as per OECD Artificial Soil) adjusted using calcium carbonate.
- Moisture:
- Water holding capacity: 44.8%
Moisture content: Soil water content was adjusted to 60% of the maximum water holding capacity. - Details on test conditions:
- TEST SYSTEM
- OECD artificial soil: Prepared according to the OECD protocol for artificial soil (1984) by mixing Sphagnum peat (10%), kaoline (20%) and quartz sand.
- Testing facility: Universitat Autonoma de Barcelona
- Test container (type, material, size): Each replicate consisted of a 1000-ml polyethylene container covered with a perforated lid allowing aeration of the test chamber.
- Amount of soil: Each vessel was filled with 500g (dry weight) or moist soil.
- No. of replicates per treatment group: 4 replicates per concentration were prepared
- Number of organisms: Ten clitellated individuals of synchronised age (4 weeks difference at most) were placed in each container.
- Feeding: Animals were fed with 5 g cooked oat flakes at the start and weekly thereafter.
- Photoperiod: Test was maintained at 16: 8 light/dark photoperiod.
- Moisture loss: The vessels were weighed weekly to check for moisture loss and weight restored with distilled water where necessary.
SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: The results presented here relate to the OECD Artificial Soil, however two natural soils were also tested. These correspond to a loamy agricultural soil (PRA) within the Universitat Autonoma de Barcelona campus (Cerdanyola del Valles, NE Spain) and the other was a loam sand soil (UAB) from the dry grassland in Serra de Prades (Tarragona, NE Spain).
- Pesticide use history at the collection site: Not relevant to the OECD Artificial Soil, however, pesticide use had ceased five years previously in the agricultural soil (UAB).
- Collection procedures: Natural soils were defaunated by alternating two consecutive freeze-thaw cycles (-20 °C for 4 days, then +20 °C for 4 days).
- Sampling depth (cm): Not relevant to the OECD soil, but natural soils were collected from the upper layer (20 cm).
- Soil texture (if natural soil): PRA soil
- % sand: 80.3
- % silt: 16.5
- % clay: 3.2
- Soil texture (if natural soil): UAB soil
- % sand: 36.4
- % silt: 44.9
- % clay: 18.7
- Composition (if artificial substrate): OECD Artificial Soil
- Organic carbon (%): 3.45
- Maximum water holding capacity (in % dry weight): 44.8% adjusted to 60% WHC
- CEC: 8.94 - Nominal and measured concentrations:
- Nominal concentrations: The test concentrations were selected following a preliminary assay consisting of 0, 1, 10, 100, 1000 and 10000 mg of nonylphenol per kg of soil. The range of concentrations showing an inhibition between 10% and 90% were selected for the study.
- Reference substance (positive control):
- no
- Duration:
- 8 wk
- Dose descriptor:
- EC10
- Effect conc.:
- 55.8 mg/kg soil dw
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Details on results:
- No differences in the outcomes between controls with and without the addition of acetone were found in any of the assays. Validity criteria were met for all species.
- Adult weight in worms at the start of the test was in the range inidicated in the Guideline (200-650 mg)
- Survival was over 90%
- Reproduction was over 30 juveiles with a coefficient of variation below 30%.
Mean values (+/- standard deviation) are provided for the controls in the different test soils below:
- Survival in adults: OECD 9.8 (±0.8); PRA 10.0 (±0.0); UAB 10.0 (±0.0)
- Reproduction in juveniles: OECD 53.4 (±14.6); PRA 73.1 (±16.2); UAB 104.2 (±26.7)
- Initial weight /worm: OECD 417 (±114); PRA 488 (±86); UAB 472 (±90)
- Final weight/worm: OECD 381 (±77); PRA 450 (±106); UAB 433 (±87) - Results with reference substance (positive control):
- No data
- Reported statistics and error estimates:
- - EC10 was calculated using Statistica 6.0 (StatSoft Inc., Tulsa USA).
- The influence of soil properties was assessed by Generalised Linear Models (Brodgar 2.0, Highland Statistics Ltd, Newburgh, UK). - Validity criteria fulfilled:
- yes
- Conclusions:
- The authors considered nonylphenol to be more toxic than the nonylphenol ethoxylate mixture and considered the results to be in agreement with existing published literature. Nonylphenol presented low phytotoxicity but moderate toxicity for invertebrates, reproduction being clearly the most sensitive endpoint. Nonylphenol generally affected the invertebrates's reproduction at lower concentrations than those affecting growth or survival. Earthworms and collembolans were more sensitive taxa compared to enchytraeids.
- Executive summary:
In an 8-week chronic toxicity study, earthworms (Eisenia andreii) were exposed at control, solvent control, and nonylphenol at a range of concentrations showing inhibition between 10% and 90% from an initial range-finding test. The EC10 based on reproduction was 55.8 mg nonylphenol/kg, using an artificial soil test medium. Results for two natural soils were also reported. Based on the results of this study, nonylphenol would be considered to have adverse acute effects to Eisenia andreii.
This study is classified as acceptable and is similar in methodology to the requirements of the ISO Guideline 11268 -2 (ISO, 1998).
Referenceopen allclose all
The LC50 is prioritised for reporting from this study as it a short-term exposure and directly comparable with the EC50 from other short-term toxicity studies (e.g., Hulzebos et al, 1993). A NOEC and a LOEC based on growth effects of 32 and 100 mg nonylphenol/kg were also reported in this study.
A summary of the physiological effects measured in the long-term Eisenia andreii reproduction study is provided below:
Substance |
Exposure concentration |
Mean number of hatched cocoons |
Mean number of unhatched cocoons |
Mean number of live worms |
Nonylphenol |
Control |
12.5+2.1 |
2.8+1.3 |
24.3+8.1 |
1.0 mg kg-1 |
13.5+4.4 |
3.0+1.4 |
23.0+10.6 |
|
3.2 mg kg-1 |
9.8+4.3 |
2.0+2.8 |
21.5+7.5 |
|
10 mg kg-1 |
13.3+2.5 |
3.0+1.6 |
22.5+4.7 |
|
32 mg kg-1 |
7.8+3.2 |
4.3+2.9 |
17.0+7.3 |
|
100 mg kg-1 |
11.0+5.3 |
2.3+1.7 |
23.0+10.0 |
Reproduction was a more sensitive endpoint than survival in the encytraeid study. By way of comparison using the OECD artificial soil, the EC10 for reproduction was 24.0 mg/kg compared with the EC10 and EC50 for survival which were 663.5 and 906.7 mg/kg respectively. When the results for the three soils were compared, enchytraeids (reproduction) in OECD Artificial Soil (EC10 of 24.0 mg/kg) were more sensitive than enchytraeids in the other soils; reproduction in the PVA soil (EC10 of 455.8 mg/kg) and UAB soil (
EC10 of 197.2 mg/kg). The results for the OECD Artificial Soil have been selected for this hazard assessment as the standard soil data are directly comparable with other studies where artificial soils were used (for example, Johnson et al, 2005).
Toxicity values of 4 -nonylphenol for several terrestrial organisms in the three different soil types:
Species | Endpoint | Soil | EC10 | EC50 |
B. rapa | Freshweight | OECD | 574.8 (279.9, 1,180.4) | 1,449.1 (785.0, 2,674.7) |
Germination | 695.9 (286.3, 1,691.6) | 8,159.2 (5,904.0, 11,276.0) | ||
L. perenne | Freshweight | OECD | 738.9 (49.6, 11,011.2) | 4,011.6 (862.2, 18,664.5) |
Germination | 1,385.8 (696.5, 2,757.3) | 7,500.7 (6,031.2, 9,328.3) | ||
E. andreii | Survival | OECD | 343.9 (160.6, 736.4) | 625.5 (427.9, 914.1) |
PRA | 182.4 (157.6, 211.2) | 290.7 (272.0, 310.7) | ||
UAB | 498.2 (439.2, 565.2) | 622.0 (592.3, 653.2) | ||
Reproduction | OECD | 55.8 (13.7, 227.7) | 82.0 (47.6, 141.1) | |
PRA | 43.1 (26.9, 69.0) | 69.6 (57.7, 83.9) | ||
UAB | 63.6 (46.5, 87.1) | 124.3 (110.5, 139.9) | ||
Biomass | OECD | 88.3 (50.0, 155.8) | 309.1 (240.4, 397.3) | |
PRA | 121.2 (93.6, 156.9) | 240.2 (218.0, 264.8) | ||
UAB | 289.4 (219.1, 382.4) | 523.2 (474.5, 576.9) | ||
E. crypticus | Survival | OECD | 663.5 (444.9, 989.5) | 906.7 (786.5, 1,045.3) |
PRA | 214.5 (171.3, 268.6) | 316.2 (284.9, 350.9) | ||
UAB | 227.0 (89.5, 575.3) | 615.2 (431.9, 876.3) | ||
Reproduction | OECD | 24.0 (5.2, 110.7) | 225.8 (134.8, 378.4) | |
PRA | 455.8 (311.2, 667.5) | 641.7 (554.2, 742.9) | ||
UAB | 197.2 (97.9, 397.3) | 212.8 (194.6, 232.8) | ||
F. candida | Survival | OECD | 102.0 (69.7, 149.3) | 138.7 (129.4, 148.7) |
PRA | 52.3 (47.0, 58.1) | 69.8 (65.8, 74.1) | ||
UAB | 89.1 (70.0, 113.5) | 116.3 (104.2, 129.8) | ||
Reproduction | OECD | 63.2 (47.3, 84.5) | 92.8 (83.8, 102.8) | |
PRA | 45.0 (27.5, 73.7) | 64.4 (54.6, 75.9) | ||
UAB | 54.8 (23.9, 125.7) | 88.0 (79.9, 96.9) |
Reproduction was a more sensitive endpoint than survival in the earthworm study. By way of comparison using the OECD artificial soil, the EC10 for reproduction was 55.8 mg/kg compared with the EC10 and EC50 for survival which were 343.9 and 625.5 mg/kg respectively. When the results for the three soils were compared, earthworms in PRA soil were more sensitive (EC10 of 43.1 mg/kg) than the OECD Artificial Soil (EC10 of 55.8 mg/kg) and UAB soil (EC10 of 63.6 mg/kg). The results for the OECD Artificial Soil have been selected for this hazard assessment as the standard soil data are directly comparable with other studies where artificial soils were used (for example, Johnson et al, 2005).
Toxicity values of 4 -nonylphenol for several terrestrial organisms in the three different soil types:
Species | Endpoint | Soil | EC10 | EC50 |
B. rapa | Freshweight | OECD | 574.8 (279.9, 1,180.4) | 1,449.1 (785.0, 2,674.7) |
Germination | 695.9 (286.3, 1,691.6) | 8,159.2 (5,904.0, 11,276.0) | ||
L. perenne | Freshweight | OECD | 738.9 (49.6, 11,011.2) | 4,011.6 (862.2, 18,664.5) |
Germination | 1,385.8 (696.5, 2,757.3) | 7,500.7 (6,031.2, 9,328.3) | ||
E. andreii | Survival | OECD | 343.9 (160.6, 736.4) | 625.5 (427.9, 914.1) |
PRA | 182.4 (157.6, 211.2) | 290.7 (272.0, 310.7) | ||
UAB | 498.2 (439.2, 565.2) | 622.0 (592.3, 653.2) | ||
Reproduction | OECD | 55.8 (13.7, 227.7) | 82.0 (47.6, 141.1) | |
PRA | 43.1 (26.9, 69.0) | 69.6 (57.7, 83.9) | ||
UAB | 63.6 (46.5, 87.1) | 124.3 (110.5, 139.9) | ||
Biomass | OECD | 88.3 (50.0, 155.8) | 309.1 (240.4, 397.3) | |
PRA | 121.2 (93.6, 156.9) | 240.2 (218.0, 264.8) | ||
UAB | 289.4 (219.1, 382.4) | 523.2 (474.5, 576.9) | ||
E. crypticus | Survival | OECD | 663.5 (444.9, 989.5) | 906.7 (786.5, 1,045.3) |
PRA | 214.5 (171.3, 268.6) | 316.2 (284.9, 350.9) | ||
UAB | 227.0 (89.5, 575.3) | 615.2 (431.9, 876.3) | ||
Reproduction | OECD | 24.0 (5.2, 110.7) | 225.8 (134.8, 378.4) | |
PRA | 455.8 (311.2, 667.5) | 641.7 (554.2, 742.9) | ||
UAB | 197.2 (97.9, 397.3) | 212.8 (194.6, 232.8) | ||
F. candida | Survival | OECD | 102.0 (69.7, 149.3) | 138.7 (129.4, 148.7) |
PRA | 52.3 (47.0, 58.1) | 69.8 (65.8, 74.1) | ||
UAB | 89.1 (70.0, 113.5) | 116.3 (104.2, 129.8) | ||
Reproduction | OECD | 63.2 (47.3, 84.5) | 92.8 (83.8, 102.8) | |
PRA | 45.0 (27.5, 73.7) | 64.4 (54.6, 75.9) | ||
UAB | 54.8 (23.9, 125.7) | 88.0 (79.9, 96.9) |
Description of key information
The Johnson et al (2005) short-term LC50 of 88.6 mg nonylphenol/kg for Eisenia andreii survival meets reliability ownership and adequecy requirements for REACH. The study uses the preferred test species and the preferred test medium, artificial soil. The long-term study of Domene et al (2009) followed international OECD and ISO protocols, uses standard test species and achieves the highest Klimisch rating. Long-term exposure of Enchytraeus crypticus reported an EC10 of 24 mg nonylphenol/kg.
Key value for chemical safety assessment
- Short-term EC50 or LC50 for soil macroorganisms:
- 88.6 mg/kg soil dw
- Long-term EC10, LC10 or NOEC for soil macroorganisms:
- 24 mg/kg soil dw
Additional information
Only one reliable study was available for short-term exposure to soil macro-organisms except arthropods and this was the Key Study by Johnson et al., (2005). The preferred earthworm species, Eisenia andreii was exposed to nonylphenol for 14 days with a reported NOEC and LOEC based on survival of 32 and 100 mg/kg, respectively. ENVIRON UK Ltd calculated an LC50of 88.6 mg nonylphenol/kg for the survival data using Probit statistical analysis. The derivation of an LC50means the macro-invertebrate data are more easily comparable with other short-term studies, for example plant study by Hulzeboset al., 1993), where macro-invertebrates appeared to be more sensitive than plants.
Two studies representing three endpoints were reported for long-term macro-invertebrate exposures to nonylphenol resulting in NOEC or EC10 values ranging from 24 to 100 mg nonylphenol/kg. Both Johnsonet al., 2005 and Domeneet al., 2009 carried out 8-week earthworm (Eisenia andreii) reproduction studies using nonylphenol spiked artificial soil exposures. The Johnsonet al., 2005 study did not record any effects on reproduction at the highest concentration tested (NOEC = 100 mg/kg) but the Domeneet al., 2009 study statistically derived a more robust EC10of 55.8 mg/kg. Domeneet al., 2009 also studied reproduction inEnchytraeus crypticusand statistically derived an EC10of 24 mg nonylphenol/kg. TheE. crypticus study duration was 4 weeks and slightly shorter in duration than the standard 6-week duration of the standard ISO Guideline but this is still considered to be acceptable as this smaller enchytraeid species’ breeding cycle was completed.
In addition, Domene et al., 2009 compared the standard toxicity tests between OECD artificial soils and two natural soils. In the earthworm tests where reproduction in the OECD was affected at the 10% level leading to an EC10of 55.8 mg/kg, the loamy sand soil from dry grassland affected reproduction at 43.1 mg/kg (more sensitive) but the agricultural loamy soil was less sensitive with an EC10of 63.6 mg/kg. However, when the enchytraeid study was compared, both natural soils reported far less sensitive values than the OECD soil; 455.8 and 197.2 mg/kg compared to the EC10of 24 mg/kg in the OECD soil. The inconsistency in trends between natural and artificial soils means that the artificial soils are preferred for use in this REACH asseessment to ensure studies are comparable where possible.
The long-term study of Domene et al., (2009) forEnchytraeus crypticuswith an EC10of 24 mg nonylphenol/kg is the Key Study for this endpoint and is supported by the less sensitive long-term and short-term earthworm studies.
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