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EC number: - | CAS number: -
- 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
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- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
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- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
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- Endpoint summary
- Stability
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- 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
![](https://poisoncentres.echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_ecotoxicological-information.png)
Long-term toxicity to aquatic invertebrates
Administrative data
Link to relevant study record(s)
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 3rd January - 17th January 2020
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: US EPA 821-R-02-013
- Version / remarks:
- USEPA (2002) Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms. Fourth Edition. October 2002
- Deviations:
- no
- GLP compliance:
- yes
- Specific details on test material used for the study:
- Naphtha (petroleum), light steam-cracked, debenzenized, C8-16-cycloalkadiene conc. (LOA Category L2-DCPD)
CAS: 68478-10-4
EC: 270-790-1
CRO ID: MRD-19-988
No test substance characterization was performed by the testing facility. The test item was fully characterised by the sponsor before submission to the laboratory. - Analytical monitoring:
- yes
- Details on sampling:
- On Day 0 (initiation) and Day 1 “old”, samples were collected from four treatment group WAFs (0.04, 0.19, 0.41 and 2.0 mg/L) and the control WAF. Day 5 “new” WAFs and Day 6 “old” were collected from three treatment group (0.04, 0.19, and 0.41 mg/L) and the control WAF. “Old” samples were collected from individual replicate test chambers. Duplicate samples were analyzed from the treatment groups collected; and a single sample was analyzed for the control group.
- Vehicle:
- no
- Details on test solutions:
- New WAFs were prepared daily by adding the appropriate volume of test substance using gas tight (glass and stainless steel) syringes to the surface of the dilution water in glass aspirator bottles. The 0.02 mg/L treatment group was prepared in 20L of dilution water in a 21.5L glass aspirator bottle, the 0.04 and 0.09 mg/L treatment groups were prepared in 12L of dilution water in 13.5L glass aspirator bottles, the 0.19 and 0.41 mg/L treatment groups were prepared in 4L of dilution water in 4.3L glass aspirator bottles and the 0.91 and 2.0 mg/L treatment groups and control were prepared in 2L of dilution water in 2.2L glass aspirator bottles. All aspirator bottles were sealed with Teflon® screw caps and placed in an environmental chamber at test temperature with a 16:8 light cycle. The volume of test substance dispensed into each aspirator bottle was calculated using the nominal loading rate, the quantity of dilution water and the specific density of the test substance. The test substance specific density was 0.956 g/mL referenced in the Safety Data Sheet.
The WAFs were mixed with Teflon® coated stir bars on magnetic stir plates with a ≤10% vortex for 24 ± 1h. At the time of mixing, all batch solutions appeared clear and colorless.
At the end of mixing, the WAFs were allowed to settle for approximately 1h ± 30 mins. After the end of the settling period, approximately 100 mL of solution was removed and discarded, then WAFs were removed from the aspirator bottles through the outlet at the bottom of the vessels and placed into the respective test chambers. All test chambers contained no headspace.
A control was prepared in the same manner without test substance. Daily renewals consisted of fresh control and treatment solutions being distributed into new test vials with the appropriate volume of feed, and the C. dubia parent being transferred from the “old” treatment vials to the “new” treatment vials. - Test organisms (species):
- Ceriodaphnia dubia
- Details on test organisms:
- C. dubia were cultured at the testing facility. The original culture was supplied by Environmental Consulting and Testing Inc. Superior, WI 54880 USA.
A total of 80 female <23-h neonates were used, all released within 4 hour period. Neonates used for this study were from adults that were 6 days old.
C. dubia were maintained in 20 mL glass scintillation vials with approximately 20 mL of reconstituted water (dilution water) supplemented with Na2SeO4 at 2μg/L Se and 1μg/L Vitamin B12 at 25 ± 2°C under a 16 hour light: 8 hour dark photoperiod. Stock cultures were transferred to fresh reconstituted water daily and fed a suspension of Pseudokirchneriella subcapitata and yeast-cereal leaves-trout mixture (YCT). The concentration of P. subcapitata and volume of feeds are documented in the culture log. Algae and YCT are supplied by Environmental Consulting and Testing Inc. Superior, WI 54880 USA.
Stock cultures of test organisms were started at least three weeks before the brood animals were needed. Documentation on survival of brood organisms and the number of offspring is maintained at the testing facility.
Suspension of P. subcapitata (190 μL of a 2.7x10^7 cells/mL) to provide approximately 2.6x10^5 cells/mL and YCT (100 μL) were added to each freshly prepared test chamber daily prior to organism transfer. - Test type:
- semi-static
- Water media type:
- other: Reconstituted Deionised
- Limit test:
- no
- Total exposure duration:
- 6 d
- Post exposure observation period:
- None
- Hardness:
- Reconstituted moderately-hard water (APHA, 2017) was prepared from UV-sterilized, deionized well water and reagent grade salts (NaHCO3, CaSO4, MgSO4, and KCl) and recommended micronutrients Na2SeO4 at 2μg/L Se and 1μg/L Vitamin B12. The hardness of the moderately-hard reconstituted water was 84 mg/L (as CaCO3). Batch water was aerated and covered to protect it from light.
Hardness was measured at beginning of the test in each concentration - Test temperature:
- Temperatures in the exposure chambers ranged from 24.7 – 26.0 ºC, and was measured at the beginning and end of each 24h exposure.
Temperature was continuously monitored in the test area using the Watchdog V5 monitoring system. During the in-life phase temperature in the environmental chamber ranged from 24.6 to 25.5°C. - pH:
- pH was measured at the beginning and end of each 24h exposure. New solution pH measurements did not vary by more than 0.38 units from the old test solution.
The protocol required that the pH should not vary by more than 0.3 units between corresponding new and old solutions however the pH varied from 0.31-0.38 units on several occasions.
pH day 0 in control was 8.26, and in the treatment groups 8.26-8.43, and day 6 control pH was 7.99 and in treated groups 7.99-8.09. - Dissolved oxygen:
- Dissolved oxygen was measured at the beginning and end of each 24h exposure. Dissolved oxygen (DO) remained at or above 6.42 mg/L throughout the test.
- Salinity:
- Not measured
- Conductivity:
- Conductivity was measured at beginning of the test in each concentration. In control it was 373.0μs/cm, and in treated groups 361.8-371.3 μs/cm.
- Nominal and measured concentrations:
- Nominal concentrations were 0 (control), 0.02, 0.04, 0.09, 0.19, 0.41, 0.91, 2.0 mg/L.
- Details on test conditions:
- Test Chamber / Organism Loading:
Test chambers were 20 mL glass scintillation vials containing one C. dubia and approximately 20 mL of test solution with no headspace. Each chamber was closed with PTFE-lined screw caps to minimize contamination, evaporation and/or volatilization.
Selection:
Test chamber position in the test area and organism assignment followed a randomized block design. The offspring from a single female were distributed evenly among the treatments and the control. Neonates were selected from third broods comprising at least eight neonates from adults that were 6 days old. The study director determined organism suitability. A printout of the randomization schedule generated using JMP v. 14.1 (JMP, 2018) was included in the raw data.
Individual C. dubia were not identified. Each test chamber was labeled with study number, treatment group, replicate, and randomization number.
Discrete Measurements:
Temperature, pH, and dissolved oxygen were measured at the beginning (before renewal) and end of each 24h exposure. Conductivity and hardness were measured at the beginning of the test in each concentration. All fresh test solutions were taken directly from the mixing vessels. Old test solutions from (2) individual replicates, if available, were composited for measurement following organism transfer and neonate counts.
Experimental Evaluation:
Observations for immobilization were performed and recorded daily at 24 ± 1h intervals after in-life initiation. Immobilization was considered the lack of swimming ability within 15 sec after gentle agitation of the test chamber. The adults were transferred via pipette to chambers containing fresh test solution daily. Neonate presence and enumeration from each adult was performed following the adult transfer on a daily basis. After completion of the study, the test organisms were discarded and monitoring of environmental conditions was discontinued.
Exposure Duration:
6 days - Key result
- Duration:
- 6 d
- Dose descriptor:
- EL10
- Effect conc.:
- 0.85 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Key result
- Duration:
- 6 d
- Dose descriptor:
- other: EL20
- Effect conc.:
- 0.89 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Key result
- Duration:
- 6 d
- Dose descriptor:
- other: NOEL
- Effect conc.:
- 0.41 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Key result
- Duration:
- 6 d
- Dose descriptor:
- other: LOEL
- Effect conc.:
- 0.91 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Details on results:
- This study was considered acceptable as there was no mortality in the control C. dubia for the duration of the study. Additionally, 100% of control C. dubia produced three broods by Day 6 (study termination). The mean number of live offspring produced per adult in the control group surviving at the end of study was 25 neonates. Guideline acceptability criteria states that 80% or greater of control organisms must survive and have an average of 15 or more young per surviving female while 60% of surviving controls must produce three broods.
Analytical verification of the control and test solution concentrations of 0.04, 0.19, 0.41, and 2.0 mg/L (if available), was performed at Day 0 and Day 5 representing “new” solutions and Day 1 and Day 6 representing “old” solutions using BE-SPME by GC-FID and reported as μmol as 2,3-dimethylnaphthalene/mL PDMS. Nominal loading rates were used in statistical endpoint evaluations.
New solution pH measurements did not vary by more than 0.38 units from the old test solution. Dissolved oxygen (DO) remained at or above 6.42 mg/L throughout the test. Temperatures in the exposure chambers ranged from 24.7 - 26.0 ºC.
No observation of test substance insolubility (surface slicks, precipitates, and adherence to the test chamber) was noted during the time of organism observations. No abnormal behavior or appearance was observed in the control or lowest six treatment groups. Lethargic organisms were observed in the 2.0 mg/L treatment group on Day 1. No observations of aborted eggs or immobilized neonates were noted in the control or treatment groups throughout the exposure. Immobilized test organisms were observed in the 2.0 mg/L treatment group, where 50% of organisms were immobilized by Day 1, and 100% of organisms were immobilized by Day 2.
Both acute (LL50) and chronic (EL10, EL20, NOEL, LOEL) endpoints were calculated for this study. The acute endpoint was calculated using 48h immobilization data and the chronic endpoints were calculated using reproduction data gathered throughout the duration of the study.
PROTOCOL DEVIATION
The protocol required that the pH should not vary by more than 0.3 units between corresponding new and old solutions however the pH varied from 0.34-0.57 units on several occasions.
No critical effects were observed in the control group and the validity of the study was maintained. - Results with reference substance (positive control):
- No chronic reference toxicity test was performed at the testing facility.
- Reported statistics and error estimates:
- The EL10 and EL20 chronic endpoints were determined based on neonate counts and nominal loading rates relative to the control value. The EL10 and EL20 values with associated confidence intervals were calculated based on a non-linear regression model, the Hill Model, using the Benchmark Dose method (USEPA, 2010).
The NOEL and LOEL chronic endpoint data was determined based on nominal loading rates and reproduction data using JMP v. 14.1 (JMP, 2018). The Shapiro-Wilk goodness of fit test was used to determine if the data was normally distributed (Shapiro and Wilk, 1965). An ANOVA (Analysis of Variance) was used to determine homogeneity of variance of reproduction data. Nonparametric comparison with control using the Steel method (Steel, 1959) was performed to determine the NOEL and LOEL of nominal loading rates vs. total number of neonates produced in three broods. This statistical analysis identified the 0.02 mg/L level as statistically significant, but due to no effect at the consecutive three higher levels, it may be interpreted that it is not justifiable as a LOEL. - Validity criteria fulfilled:
- yes
- Conclusions:
- Chronic (EL10, EL20, NOEL, LOEL) endpoints were calculated for this study. The chronic endpoints were calculated using reproduction data gathered throughout the 6 days of the study and were based on nominal rates:
EL10: 0.85 (97.5% confidence interval of 0.50-1.20) mg/L
EL20: 0.89 (97.5% confidence interval of 0.70 -1.07) mg/L
NOEL: 0.41 mg/L
LOEL: 0.91 mg/L
Data above is based on nominal loading rates
*Values are 97.5% confidence intervals
**Values are 99% confidence intervals - Executive summary:
This study was conducted for the Sponsor to evaluate the effects of the water accommodated fractions (WAFs) of Naphtha (petroleum), light steam-cracked, debenzenized, C8-16-cycloalkadiene conc. (CAS: 68478-10 -4) on survival and reproductive output of the cladoceran, Ceriodaphnia dubia, in a 3-brood semi-static dose-response test.
WAFs were prepared daily for each treatment group by adding the appropriate amount of the test substance using gas tight (glass and stainless steel) syringes to moderately hard reconstituted water with micronutrients in glass aspirator bottles sealed with PTFE screw caps. The WAFs were held in an environmental chamber with a 16:8 light cycle at approximately test temperature, and were mixed with Teflon-coated stir bars on magnetic stir plates for approximately 24 ± 1h. The control WAF was prepared in the same manner without test substance addition. At the end of mixing, the WAFs were allowed to settle for approximately 1h ± 30 mins. At the end of each settling period, approximately 100 mL of the WAF solution was removed and discarded, then solutions were removed from the mixing vessels through the outlet at the bottom of the vessels and placed into the respective test chambers. All test chambers contained no headspace. The WAF loading rates tested were control (0), 0.02, 0.04, 0.09, 0.19, 0.41, 0.91 and 2.0 mg/L.
Analytical verification of the test solutions for 0.04, 0.19, 0.41 and 2 mg/L and the control were performed on Day 0 and Day 5 “new” solutions and Day 1 and Day 6 “old” solutions, if available, using the automated biomimetic extraction technique employing solid phase micro-extraction (BE-SPME). Nominal loading rates were used in statistical endpoint evaluations.
Chronic (EL10, EL20, NOEL, LOEL) endpoints were calculated for this study. The chronic endpoints were calculated using reproduction data gathered throughout the 6 days of the study and were based on nominal rates:
EL10: 0.85 (97.5% confidence interval of 0.50-1.20) mg/L
EL20: 0.89 (97.5% confidence interval of 0.70 -1.07) mg/L
NOEL: 0.41 mg/L
LOEL: 0.91 mg/L
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2 July 2019 - 11 July 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: US EPA 821-R-02-13
- Version / remarks:
- USEPA (2002) Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms. Fourth Edition. October 2002
- Deviations:
- yes
- Remarks:
- No chronic reference toxicity test was performed at the testing facility
- GLP compliance:
- yes
- Specific details on test material used for the study:
- Solvent naphtha (coal)
CAS: 65996-79-4
CRO ID: MRD-19-990
EC: 266-013-0
No test substance characterization was performed by the testing facility. The test item was fully characterised by the sponsor before submission to the laboratory. - Analytical monitoring:
- yes
- Details on sampling:
- On Day 0 (initiation) and Day 5, samples were collected from four treatment group WAFs (0.32, 1.25, 2.5 and 10 mg/L) and the control WAF representing “new” solutions and on Days 1 and 6 representing “old” solutions. Old samples were collected from individual replicates for analysis. Duplicate samples were analyzed from the treatment groups collected; and a single sample was analyzed for the control group. All samples (no headspace) were collected in clear 20 mL LEAP glass vials with PTFE septa screw caps and analyzed the day they were sampled or refrigerated until analysis. All samples retained were stored under refrigerated conditions.
- Vehicle:
- no
- Details on test solutions:
- New individual WAFs were prepared daily by adding the appropriate volume of test substance using gas tight (glass and stainless steel) syringes to dilution water in glass aspirator bottles. For the ease of dosing at ~1.0 µl and greater, the 0.16 mg/L treatment group was prepared in 20L of dilution water in a 21.5L glass aspirator bottle, the 0.32, 0.63 and 1.25 mg/L treatment groups were prepared in 12L of dilution water in 13.5L glass aspirator bottles, the 2.5 mg/L treatment group was prepared in 4L of dilution water in 4.3L glass aspirator bottles and the 5.0 and 10.0 mg/L treatment groups and control were prepared in 2L of dilution water in 2.2L glass aspirator bottles. All aspirator bottles were sealed with Teflon® screw caps. The volume of test substance dispensed into each aspirator bottle was calculated using the nominal loading rate, the quantity of dilution water and the specific density of the test substance. The test substance specific density was 0.95 g/mL referenced in the Safety Data Sheet.
The WAFs were mixed with Teflon® coated stir bars on magnetic stir plates with a ≤10% vortex for an average of 23.4 h at room temperature. At the time of mixing, all batch solutions appeared clear and colorless with the exception of the 10 mg/L treatment which had visible test substance on the surface of the WAF.
At the end of mixing, the WAFs were allowed to settle for approximately 1h. All batch solutions appeared clear and colorless with the exception of the 10 mg/L which contained visible test substance on the surface of the WAF. After the end of the settling period, approximately 100 mL of solution was removed and discarded, then solutions were removed from the aspirator bottles through the outlet at the bottom of the vessels and placed into the respective test chambers. All test chambers contained no headspace.
A control was prepared in the same manner without test substance. Daily renewals consisted of fresh control and treatment solutions being distributed into new test vials with the appropriate volume of feed, and the C. dubia parent being transferred from the “old” treatment vials to the “new” treatment vials. - Test organisms (species):
- Ceriodaphnia dubia
- Details on test organisms:
- C. dubia were cultured at the testing facility. The original culture was supplied by Environmental Consulting and Testing Inc. Superior, WI 54880 USA.
C. dubia were maintained in 20 mL glass scintillation vials with approximately 20 mL of reconstituted water (dilution water) supplemented with Na2SeO4 at 2µg/L Se and 1µg/L Vitamin B12 at 25 ± 2°C under a 16 hour light: 8 hour dark photoperiod. Stock cultures were transferred to fresh reconstituted water daily and fed a suspension of Pseudokirchneriella subcapitata and yeast-cereal leaves-trout mixture (YCT). The concentration of P. subcapitata and volume of feeds are documented in the culture log. Algae and YCT are supplied by Environmental Consulting and Testing Inc. Superior, WI 54880 USA.
Stock cultures of test organisms were started at least three weeks before the brood animals were needed. Documentation on survival of brood organisms and the number of offspring is maintained at the testing facility
A total of 80 female <24-h neonates were used, all released within ~ 5 h period. The neonates used for this study were from adults that were 6 days old.
Suspension of P. subcapitata (170 µL of a 2.93x107 cells/mL) to provide approximately
2.49x105 cells/mL and YCT (100 µL) were added to each freshly prepared test chamber daily prior to organism transfer. - Test type:
- semi-static
- Water media type:
- other: Reconstituted Deionised
- Limit test:
- no
- Total exposure duration:
- 7 d
- Post exposure observation period:
- None
- Hardness:
- Reconstituted moderately-hard water (APHA, 2017) was prepared from UV-sterilized, deionized well water and reagent grade salts (NaHCO3, CaSO4, MgSO4, and KCl) and recommended micronutrients Na2SeO4 at 2μg/L Se and 1μg/L Vitamin B12. The hardness of the moderately-hard reconstituted water was 96 mg/L (as CaCO3). Batch water was aerated and covered to protect it from light.
Hardness was measured at beginning of the test in each concentration - Test temperature:
- Temperatures in the exposure chambers ranged from 24.0 – 25.5 ºC, and was measured at the beginning and end of each 24h exposure.
Temperature was continuously monitored in the test area using the Watchdog V5 monitoring system. During the in-life phase temperature in the environmental chamber ranged from 24.8 to 25.5 ºC. - pH:
- pH was measured at the beginning and end of each 24h exposure. New solution pH measurements did not vary by more than 0.45 units from the old test solution.
The protocol required that the pH should not vary by more than 0.3 units between corresponding new and old solutions however the pH varied from 0.31-0.45 units on several occasions.
pH day 0 in control was 8.11, and in the treatment groups 8.26-8.29, and day 7 control pH was 7.69 and in treated groups 7.84-8.00. - Dissolved oxygen:
- Dissolved oxygen was measured at the beginning and end of each 24h exposure. Dissolved oxygen (DO) remained at or above 6.15 mg/L throughout the test.
- Salinity:
- Not measured
- Conductivity:
- Conductivity was measured at beginning of the test in each concentration. In control it was 365.0μs/cm, and in treated groups 362.1-369.0μs/cm.
- Nominal and measured concentrations:
- Nominal concentrations: 0 (control), 0.16, 0.32, 0.63, 1.25, 2.5, 5.0, 10 mg/L.
- Details on test conditions:
- Test Chamber / Organism Loading:
Test chambers were 20 mL glass scintillation vials containing one C. dubia and approximately 20 mL of test solution with no headspace. Each chamber was closed with PTFE-lined screw caps to minimize contamination, evaporation and/or volatilization.
Selection:
Test chamber position in the test area and organism assignment followed a randomized block design. The offspring from a single female were distributed evenly among the treatments and the control. Neonates were selected from third broods comprising at least eight neonates from adults that were 6 days old. The study director determined organism suitability. A printout of the randomization schedule generated using JMP v. 14.1 (JMP, 2018) was included in the raw data.
Individual C. dubia were not identified. Each test chamber was labeled with study number, treatment group, replicate, and randomization number.
Discrete Measurements:
Temperature, pH, and dissolved oxygen were measured at the beginning (before renewal) and end of each 24h exposure. Conductivity and hardness were measured at the beginning of the test in each concentration. All fresh test solutions were taken directly from the mixing vessels. Old test solutions from (2) individual replicates, if available, were composited for measurement following organism transfer and neonate counts.
Experimental Evaluation:
Observations for immobilization were performed and recorded daily at 24 ± 1h intervals after in-life initiation. Immobilization was considered the lack of swimming ability within 15 sec after gentle agitation of the test chamber. The adults were transferred via pipette to chambers containing fresh test solution daily. Neonate presence and enumeration from each adult was performed following the adult transfer on a daily basis. After completion of the study, the test organisms were discarded and monitoring of environmental conditions was discontinued.
Exposure Duration:
6 days - Key result
- Duration:
- 7 d
- Dose descriptor:
- EL10
- Effect conc.:
- 0.81 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Details on results:
- This study was considered acceptable as there was no mortality in the control C. dubia for the duration of the study. Additionally, 100% of control C. dubia produced three broods by Day 7 (study termination). The mean number of live offspring produced per adult in the control group surviving at the end of study was 24 neonates. Guideline acceptability criteria states that 80% or greater of control organisms must survive and have an average of 15 or more young per surviving female while 60% of surviving controls must produce three broods.
Analytical verification of the test solution concentrations for the 0.32, 1.25, 2.50 and 10 mg/L was performed at Day 0 and Day 5 representing “new” solutions and Day 1 and Day 6 representing “old” solutions using BE-SPME by GC-FID (µmol as 2,3-dimethylnaphthalene/mL PDMS). The initial (Days 0 and 5 “New”) mean measured concentrations were 0, 1.03, 4.51, 8.23 and 29.3 (Day 0) and 0, 1.66, 4.64 and 8.20 (Day 5) µmol as 2,3-dimethylnaphthalene/mL PDMS for the control (0) 0.32, 1.25, 2.5 and 10 mg/L groups. The old (Days 1 and 6 “old”) mean measured concentrations were 0, 1.21, 3.75, 6.74 and 26.4 (Day 1) and 0, 1.05, 3.93 and 6.82 (Day 6) µmol as 2,3-dimethylnaphthalene/mL PDMS for the control (0) 0.32, 1.25, 2.5 and 10 mg/L groups. Analysis was not conducted at the 10 mg/L loading rate on Days 5 or 6 as all adult Daphnids in this group were immobilized by this point. Nominal loading rates were used in statistical endpoint evaluations.
New solution pH measurements did not vary by more than 0.45 units from the old test solution. Dissolved oxygen (DO) remained at or above 6.15 mg/L throughout the test. Temperatures in the exposure chambers ranged from 24.0 - 25.5 ºC.
No observation of test substance insolubility (surface slicks, precipitates, and adherence to the test chamber) was noted during the time of organism observations. No abnormal behavior or appearance was observed in the control or lowest five treatment groups. No observations of aborted eggs were noted in the control or treatment groups throughout the exposure. Immobilized neonates were observed on Day 5 in 1.25 and 2.5 mg/L treatment groups and on Day 6 in the 1.25 mg/L treatment. There was complete immobilization in the 5.0 mg/L group on Day 2 and 100% immobilization in the 10 mg/L group on Day 1.
Both acute (EL50) and chronic (EL10) endpoints were calculated for this study. The acute endpoint was calculated using 48h immobilization data and the chronic endpoint was calculated using reproduction data gathered throughout the duration of the study.
PROTOCOL DEVIATION
The protocol required that the pH should not vary by more than 0.3 units between corresponding new and old solutions however the pH varied from 0.34-0.57 units on several occasions.
No critical effects were observed in the control group and the validity of the study was maintained. - Results with reference substance (positive control):
- No chronic reference toxicity test was performed at the testing facility
- Reported statistics and error estimates:
- The EL10 chronic endpoint was determined based on percent reproduction inhibition and nominal concentration relative to the control values. The EL10 value with associated confidence intervals was calculated based on a non-linear regression calculation, Logistic 3P (Ratkowsky, 1990).
All acute and chronic endpoints were evaluated using JMP v.14 (JMP, 2018). - Validity criteria fulfilled:
- yes
- Conclusions:
- Chronic (EL10) endpoints were calculated for this study. The chronic endpoints were calculated using reproduction data gathered throughout the 6 days of the study and were based on nominal rates:
EL10 = 0.81 (95% confidence interval of 0.52 - 1.09) mg/L. - Executive summary:
This study was conducted for the Sponsor to evaluate the effects of the water accommodated fractions (WAFs) of Solvent naphtha (coal) (CAS: 65996-79-4) on survival and reproductive output of the cladoceran,Ceriodaphnia dubia,in a 3-brood semi-static dose-response test.
WAFs were prepared daily for each treatment group by adding the appropriate amount of the test substance using gas tight (glass and stainless steel) syringes to moderately hard reconstituted water with micronutrients in glass aspirator bottles sealed with PTFE screw caps. The batch solutions weremixed for approximately 24h[BCS1] at room temperature with Teflon-coated stir bars on magnetic stir plates. The control batch solution was prepared in the same manner without test substance addition. At the end of mixing, the solutions were allowed to settle for approximately 1h. At the end of each settling period, approximately100 mL of the WAF solution was removed and discarded, then solutions were removed from the mixing vessels through the outlet at the bottom of the vessels and placed into the respective test chambers. All test chambers contained no headspace. The WAF loading rates tested werecontrol (0), 0.16, 0.32, 0.63, 1.25, 2.5, 5.0 and 10 mg/L.
Analytical verification of the test solution concentrations 0.32, 1.25. 5.0 and 10 mg/L and the control were performed on Day 0 and Day 5 “new” solutions and Day 1 and Day 6 “old” solutions using the automated biomimetic extraction technique employing solid phase micro-extraction (BE-SPME). Nominal loading rates were used in statistical endpoint evaluations.
Chronic (EL10) endpoints were calculated for this study. The chronic endpoints were calculated using reproduction data gathered throughout the 6 days of the study and were based on nominal rates:
EL10 = 0.81 (95% confidence interval of 0.52 - 1.09) mg/L.
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 5th September 2019 - 17 September 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: EPA 821-R-02-13
- Version / remarks:
- USEPA (2002) Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms. Fourth Edition. October 2002
- Deviations:
- yes
- Remarks:
- No chronic reference toxicity test was performed at the testing facility
- GLP compliance:
- yes
- Specific details on test material used for the study:
- Distillates (petroleum), cracked, ethylene manuf. by-product, C9-10 fraction (LOA Category L2 - DCPD)
CAS: 94733-07-0
CRO ID: MRD-19-982
EC: 305-586-4
No test substance characterization was performed by the testing facility. The test item was fully characterised by the sponsor before submission to the laboratory. - Analytical monitoring:
- yes
- Details on sampling:
- On Day 0 (initiation) and Day 1 “old”, samples were collected from four treatment group WAFs (0.05, 0.32, 0.80 and 5.0 mg/L) and the control WAF. Day 5 “new” WAFs and Day 6 “old” were collected from three treatment groups (0.05, 0.32, and 0.80 mg/L) and the control WAF. “Old” samples were collected from individual replicate test chambers. Duplicate samples were analyzed from the treatment groups collected; and a single sample was analyzed for the control group.
- Vehicle:
- no
- Details on test solutions:
- New WAFs were prepared daily by adding the appropriate volume of test substance using gas tight (glass and stainless steel) syringes to the surface of the dilution water in glass aspirator bottles. The 0.02 and the 0.05 mg/L treatment groups were prepared in 20L of dilution water in a 21.5L glass aspirator bottle, the 0.13mg/L treatment group was prepared in 12L of dilution water in 13.5L glass aspirator bottles, the 0.32 and 0.80 mg/L treatment groups were prepared in 4L of dilution water in 4.3L glass aspirator bottles and the 2.0 and 5.0 mg/L treatment groups and control were prepared in 2L of dilution water in 2.2L glass aspirator bottles. All aspirator bottles were sealed with Teflon® screw caps and placed in an environmental chamber at approximately test temperature with a 16:8 light cycle. The volume of test substance dispensed into each aspirator bottle was calculated using the nominal loading rate, the quantity of dilution water and the specific density of the test substance. The test substance specific density was 0.91g/mL referenced in the Safety Data Sheet.
The WAFs were mixed with Teflon® coated stir bars on magnetic stir plates with a ≤10% vortex for an average of 23.5 h. At the time of mixing, all batch solutions appeared clear and colorless with the exception of the 5 mg/L treatment which had visible test substance on the surface of the WAF on Day 0 for approximately 1 h and Days 1 and 2.
At the end of mixing, the WAFs were allowed to settle for approximately 1h. All batch solutions appeared clear and colorless with the exception of the 10 mg/L which contained visible test substance on the surface of the WAF. After the end of the settling period, approximately 100 mL of solution was removed and discarded, then solutions were removed from the aspirator bottles through the outlet at the bottom of the vessels and placed into the respective test chambers. All test chambers contained no headspace. - Test organisms (species):
- Ceriodaphnia dubia
- Details on test organisms:
- C. dubia were cultured at the testing facility. The original culture was supplied by Environmental Consulting and Testing Inc. Superior, WI 54880 USA.
A total of 80 female <24-h neonates were used, all released within 5 hour period. Neonates used for this study were from adults that were 8 days old.
C. dubia were maintained in 20 mL glass scintillation vials with approximately 20 mL of reconstituted water (dilution water) supplemented with Na2SeO4 at 2μg/L Se and 1μg/L Vitamin B12 at 25 ± 2°C under a 16 hour light: 8 hour dark photoperiod. Stock cultures were transferred to fresh reconstituted water daily and fed a suspension of Pseudokirchneriella subcapitata and yeast-cereal leaves-trout mixture (YCT). The concentration of P. subcapitata and volume of feeds are documented in the culture log. Algae and YCT are supplied by Environmental Consulting and Testing Inc. Superior, WI 54880 USA.
Stock cultures of test organisms were started at least three weeks before the brood animals were needed. Documentation on survival of brood organisms and the number of offspring is maintained at the testing facility.
Suspension of P. subcapitata (190 μL of a 2.71x107 cells/mL) to provide approximately
2.6x105 cells/mL and YCT (100 μL) were added to each freshly prepared test chamber daily prior to organism transfer. - Test type:
- semi-static
- Water media type:
- other: Reconstituted Deionised
- Limit test:
- no
- Total exposure duration:
- 7 d
- Post exposure observation period:
- None
- Hardness:
- Reconstituted moderately-hard water (APHA, 2017) was prepared from UV-sterilized, deionized well water and reagent grade salts (NaHCO3, CaSO4, MgSO4, and KCl) and recommended micronutrients Na2SeO4 at 2μg/L Se and 1μg/L Vitamin B12. The hardness of the moderately-hard reconstituted water was 88 mg/L (as CaCO3). Batch water was aerated and covered to protect it from light.
Hardness was measured at beginning of the test in each concentration - Test temperature:
- Temperatures in the exposure chambers ranged from 22.3 – 25.3 ºC, and was measured at teh beginning and end of each 24h exposure.
Temperature was continuously monitored in the test area using the Watchdog V5 monitoring system. During the in-life phase temperature in the environmental chamber ranged from 24.2 to 25.4 ºC. - pH:
- pH was measured at the beginning and end of each 24h exposure. New solution pH measurements did not vary by more than 0.53 units from the old test solution.
The protocol required that the pH should not vary by more than 0.3 units between corresponding new and old solutions however the pH varied from 0.31-0.53 units on several occasions.
pH day 0 in control was 8.12, and in the treatment groups 8.27-8.34, and day 7 control pH was 7.66 and in treated groups 7.51-7.88. - Dissolved oxygen:
- Dissolved oxygen was measured at the beginning and end of each 24h exposure. Dissolved oxygen (DO) remained at or above 6.25 mg/L throughout the test.
- Salinity:
- Not measured
- Conductivity:
- Conductivity was measured at beginning of the test in each concentration. In control it was 385.4μs/cm, and in treated groups 379.6-382.7μs/cm.
- Nominal and measured concentrations:
- Nominal concentrations: 0 (control), 0.02, 0.05, 0.13, 0.32, 0.80, 2.0, 5.0 mg/L.
- Details on test conditions:
- Test Chamber / Organism Loading:
Test chambers were 20 mL glass scintillation vials containing one C. dubia and approximately 20 mL of test solution with no headspace. Each chamber was closed with PTFE-lined screw caps to minimize contamination, evaporation and/or volatilization.
Selection:
Test chamber position in the test area and organism assignment followed a randomized block design. The offspring from a single female were distributed evenly among the treatments and the control. Neonates were selected from third broods comprising at least eight neonates from adults that were 6 days old. The study director determined organism suitability. A printout of the randomization schedule generated using JMP v. 14.1 (JMP, 2018) was included in the raw data.
Individual C. dubia were not identified. Each test chamber was labeled with study number, treatment group, replicate, and randomization number.
Discrete Measurements:
Temperature, pH, and dissolved oxygen were measured at the beginning (before renewal) and end of each 24h exposure. Conductivity and hardness were measured at the beginning of the test in each concentration. All fresh test solutions were taken directly from the mixing vessels. Old test solutions from (2) individual replicates, if available, were composited for measurement following organism transfer and neonate counts.
Experimental Evaluation:
Observations for immobilization were performed and recorded daily at 24 ± 1h intervals after in-life initiation. Immobilization was considered the lack of swimming ability within 15 sec after gentle agitation of the test chamber. The adults were transferred via pipette to chambers containing fresh test solution daily. Neonate presence and enumeration from each adult was performed following the adult transfer on a daily basis. After completion of the study, the test organisms were discarded and monitoring of environmental conditions was discontinued.
Exposure Duration:
6 days - Key result
- Duration:
- 7 d
- Dose descriptor:
- EL10
- Effect conc.:
- 0.57 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Details on results:
- This study was considered acceptable as there was no mortality in the control C. dubia for the duration of the study. Additionally, 100% of control C. dubia produced three broods by Day 7 (study termination). The mean number of live offspring produced per adult in the control group surviving at the end of study was 29 neonates. Guideline acceptability criteria states that 80% or greater of control organisms must survive and have an average of 15 or more young per surviving female while 60% of surviving controls must produce three broods.
Analytical verification of the test solution concentrations for the 0.05, 0.32, 0.80 and 5.0 mg/L was performed at Day 0 and Day 5 representing “new” solutions and Day 1 and Day 6 representing “old” solutions using BE-SPME by GC-FID and reported as µmol as 2,3-dimethylnaphthalene/mL PDMS. Nominal loading rates were used in statistical endpoint evaluations.
New solution pH measurements did not vary by more than 0.53 units from the old test solution. Dissolved oxygen (DO) remained at or above 6.25 mg/L throughout the test. Temperatures in the exposure chambers ranged from 22.3 - 25.3 ºC.
No observation of test substance insolubility (surface slicks, precipitates, and adherence to the test chamber) was noted during the time of organism observations. No abnormal behavior or appearance was observed in the control or lowest five treatment groups. No observations of aborted eggs or immobilized neonates were noted in the control or treatment groups throughout the exposure. Immobilized ceriodaphnids were observed on Day 1 in 2.0 and 5.0 mg/L treatment groups. There was 20% immobilization in the 2.0 mg/L group by Day 4 and 100% immobilization in the 5 mg/L group on Day 2.
Chronic (EL10) endpoints were calculated for this study. The chronic endpoint was calculated using reproduction data gathered throughout the duration of the study
PROTOCOL DEVIATION
The protocol required that the pH should not vary by more than 0.3 units between corresponding new and old solutions however the pH varied from 0.34-0.57 units on several occasions.
No critical effects were observed in the control group and the validity of the study was maintained. - Results with reference substance (positive control):
- No chronic reference toxicity test was performed at the testing facility.
- Reported statistics and error estimates:
- The EL10 chronic endpoint was determined based on percent reproduction inhibition and nominal concentration relative to the control values. The EL10 value with associated confidence intervals was calculated based on a non-linear regression calculation, Logistic 4P (Ratkowsky, 1990).
All acute and chronic endpoints were evaluated using JMP v.14 (JMP, 2018). - Validity criteria fulfilled:
- yes
- Conclusions:
- Chronic (EL10) endpoints were calculated for this study.The chronic endpoints were calculated using reproduction data gathered throughout the 6 days of the study and were based on nominal rates:
EL10: 0.57 (95% confidence interval of 0.42-0.72) mg/L. - Executive summary:
This study was conducted for the Sponsor to evaluate the effects of the water accommodated fractions (WAFs) of Distillates (petroleum), cracked, ethylene manuf. by-product, C9-10 fraction (CAS: 94733-07-0) on survival and reproductive output of the cladoceran, Ceriodaphnia dubia, in a 3-brood semi-static dose-response test.
WAFs were prepared daily for each treatment group by adding the appropriate amount of the test substance using gas tight (glass and stainless steel) syringes to moderately hard reconstituted water with micronutrients in glass aspirator bottles sealed with PTFE screw caps. The WAFs were mixed in an environmental chamber at approximately at test temperature and 16:8 light cycle for 23.5h with Teflon-coated stir bars on magnetic stir plates. The control WAF was prepared in the same manner without test substance addition. At the end of mixing, the solutions were allowed to settle for approximately 1h. At the end of each settling period, approximately100 mL of the WAF solution was removed and discarded, then solutions were removed from the mixing vessels through the outlet at the bottom of the vessels and placed into the respective test chambers. All test chambers contained no headspace. The WAF loading rates tested werecontrol (0), 0.02, 0.05, 0.13, 0.32, 0.80, 2.0, and 5 mg/L.
Analytical verification of the test solution concentrations 0.05, 0.32. 0.80 and 5 mg/L and the control were performed on Day 0 and Day 5 “new” solutions and Day 1 and Day 6 “old” solutions using the automated biomimetic extraction technique employing solid phase micro-extraction (BE-SPME). Nominal loading rates were used in statistical endpoint evaluations.
Chronic (EL10) endpoints were calculated for this study.The chronic endpoints were calculated using reproduction data gathered throughout the 6 days of the study and were based on nominal rates:
EL10: 0.57 (95% confidence interval of 0.42-0.72) mg/L.
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 30th September 2019 - 10th October 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: US EPA 821-R-02-013
- Version / remarks:
- USEPA (2002) Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms. Fourth Edition. October 2002
- Deviations:
- yes
- Remarks:
- No chronic reference toxicity test was performed at the testing facility
- GLP compliance:
- yes
- Specific details on test material used for the study:
- Naphtha (petroleum), steam-cracked middle arom.
CAS: 68516-20-1
CRO ID: MRD-19-989
EC: 271-138-9
No test substance characterization was performed by the testing facility. The test item was fully characterised by the sponsor before submission to the laboratory. - Analytical monitoring:
- yes
- Details on sampling:
- On Day 0 (initiation) and Day 1 “old”, samples were collected from four treatment group WAFs (0.05, 0.32, 0.80 and 5.0 mg/L) and the control WAF. Day 5 “new” WAFs and Day 6 “old” were collected from three treatment group (0.05, 0.32, and 0.80 mg/L) and the control WAF. “Old” samples were collected from individual replicate test chambers. Duplicate samples were analyzed from the treatment groups collected; and a single sample was analyzed for the control group.
- Vehicle:
- no
- Details on test solutions:
- New WAFs were prepared daily by adding the appropriate volume of test substance using gas tight (glass and stainless steel) syringes to the surface of the dilution water in glass aspirator bottles. The 0.02 and the 0.05 mg/L treatment groups were prepared in 20L of dilution water in 21.5L glass aspirator bottles, the 0.13 mg/L treatment group was prepared in 12L of dilution water in a 13.5L glass aspirator bottle, the 0.32 and 0.80 mg/L treatment groups were prepared in 4L of dilution water in 4.3L glass aspirator bottles and the 2.0 and 5.0 mg/L treatment groups and control were prepared in 2L of dilution water in 2.2L glass aspirator bottles. All aspirator bottles were sealed with Teflon® screw caps and placed in an environmental chamber at test temperature with a 16:8 light cycle. The volume of test substance dispensed into each aspirator bottle was calculated using the nominal loading rate, the quantity of dilution water and the specific density of the test substance. The test substance specific density was 0.905 g/mL referenced in the Safety Data Sheet.
The WAFs were mixed with Teflon® coated stir bars on magnetic stir plates with a ≤10% vortex for 24 ± 1h. At the time of mixing, all batch solutions appeared clear and colorless.
At the end of mixing, the WAFs were allowed to settle for approximately 1h ± 30 mins. After the end of the settling period, approximately 100 mL of solution was removed and discarded, then WAFs were removed from the aspirator bottles through the outlet at the bottom of the vessels and placed into the respective test chambers. All test chambers contained no headspace.
A control was prepared in the same manner without test substance. Daily renewals consisted of fresh control and treatment solutions being distributed into new test vials with the appropriate volume of feed, and the C. dubia parent being transferred from the “old” treatment vials to the “new” treatment vials.
Individual C. dubia were not identified. Each test chamber was labeled with study number, treatment group, replicate, and randomization number. - Test organisms (species):
- Ceriodaphnia dubia
- Details on test organisms:
- C. dubia were cultured at the testing facility. The original culture was supplied by Environmental Consulting and Testing Inc. Superior, WI 54880 USA.
A total of 80 female <24-h neonates were used, all released within 5 hour period. Neonates used for this study were from adults that were 8 days old.
C. dubia were maintained in 20 mL glass scintillation vials with approximately 20 mL of reconstituted water (dilution water) supplemented with Na2SeO4 at 2μg/L Se and 1μg/L Vitamin B12 at 25 ± 2°C under a 16 hour light: 8 hour dark photoperiod. Stock cultures were transferred to fresh reconstituted water daily and fed a suspension of Pseudokirchneriella subcapitata and yeast-cereal leaves-trout mixture (YCT). The concentration of P. subcapitata and volume of feeds are documented in the culture log. Algae and YCT are supplied by Environmental Consulting and Testing Inc. Superior, WI 54880 USA.
Stock cultures of test organisms were started at least three weeks before the brood animals were needed. Documentation on survival of brood organisms and the number of offspring is maintained at the testing facility.
Suspension of P. subcapitata (190 μL of a 2.71x107 cells/mL) to provide approximately 2.6x105 cells/mL and YCT (100 μL) were added to each freshly prepared test chamber daily prior to organism transfer. - Test type:
- semi-static
- Water media type:
- other: Reconstituted Deionised
- Limit test:
- no
- Total exposure duration:
- 6 d
- Post exposure observation period:
- None
- Hardness:
- Reconstituted moderately-hard water (APHA, 2017) was prepared from UV-sterilized, deionized well water and reagent grade salts (NaHCO3, CaSO4, MgSO4, and KCl) and recommended micronutrients Na2SeO4 at 2μg/L Se and 1μg/L Vitamin B12. The hardness of the moderately-hard reconstituted water was 80 mg/L (as CaCO3). Batch water was aerated and covered to protect it from light.
Hardness was measured at beginning of the test in each concentration - Test temperature:
- Temperatures in the exposure chambers ranged from 24.0 – 26.0 ºC, and was measured at the beginning and end of each 24h exposure.
Temperature was continuously monitored in the test area using the Watchdog V5 monitoring system. During the in-life phase temperature in the environmental chamber ranged from 25.1 to 25.7 ºC. - pH:
- pH was measured at the beginning and end of each 24h exposure. New solution pH measurements did not vary by more than 0.57 units from the old test solution.
The protocol required that the pH should not vary by more than 0.3 units between corresponding new and old solutions however the pH varied from 0.34-0.57 units on several occasions.
pH day 0 in control was 8.34, and in the treatment groups 8.33-8.41, and day 6 control pH was 7.87 and in treated groups 7.81-8.05. - Dissolved oxygen:
- Dissolved oxygen was measured at the beginning and end of each 24h exposure. Dissolved oxygen (DO) remained at or above 6.50 mg/L throughout the test.
- Salinity:
- Not measured
- Conductivity:
- Conductivity was measured at beginning of the test in each concentration. In control it was 347.1μs/cm, and in treated groups 343.8-347.9μs/cm.
- Nominal and measured concentrations:
- Nominal concentrations: 0 (control), 0.05, 0.32, 0.8, and 5.0 mg/L.
- Details on test conditions:
- Test Chamber / Organism Loading:
Test chambers were 20 mL glass scintillation vials containing one C. dubia and approximately 20 mL of test solution with no headspace. Each chamber was closed with PTFE-lined screw caps to minimize contamination, evaporation and/or volatilization.
Selection:
Test chamber position in the test area and organism assignment followed a randomized block design. The offspring from a single female were distributed evenly among the treatments and the control. Neonates were selected from third broods comprising at least eight neonates from adults that were 6 days old. The study director determined organism suitability. A printout of the randomization schedule generated using JMP v. 14.1 (JMP, 2018) was included in the raw data.
Individual C. dubia were not identified. Each test chamber was labeled with study number, treatment group, replicate, and randomization number.
Discrete Measurements:
Temperature, pH, and dissolved oxygen were measured at the beginning (before renewal) and end of each 24h exposure. Conductivity and hardness were measured at the beginning of the test in each concentration. All fresh test solutions were taken directly from the mixing vessels. Old test solutions from (2) individual replicates, if available, were composited for measurement following organism transfer and neonate counts.
Experimental Evaluation:
Observations for immobilization were performed and recorded daily at 24 ± 1h intervals after in-life initiation. Immobilization was considered the lack of swimming ability within 15 sec after gentle agitation of the test chamber. The adults were transferred via pipette to chambers containing fresh test solution daily. Neonate presence and enumeration from each adult was performed following the adult transfer on a daily basis. After completion of the study, the test organisms were discarded and monitoring of environmental conditions was discontinued.
Exposure Duration:
6 days - Key result
- Duration:
- 6 d
- Dose descriptor:
- EL10
- Effect conc.:
- 0.49 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Key result
- Duration:
- 6 d
- Dose descriptor:
- other: EL20
- Effect conc.:
- 0.72 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Key result
- Duration:
- 6 d
- Dose descriptor:
- other: NOEL
- Effect conc.:
- 0.32 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Key result
- Duration:
- 6 d
- Dose descriptor:
- other: LOEL
- Effect conc.:
- 0.9 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Details on results:
- This study was considered acceptable as there was no mortality in the control C. dubia for the duration of the study. Additionally, 100% of control C. dubia produced three broods by Day 6 (study termination). The mean number of live offspring produced per adult in the control group surviving at the end of study was 27 neonates. Guideline acceptability criteria states that 80% or greater of control organisms must survive and have an average of 15 or more young per surviving female while 60% of surviving controls must produce three broods.
Analytical verification of the test solution concentrations for the 0.05, 0.32, 0.80 and 5.0 mg/L was performed at Day 0 and Day 5 representing “new” solutions and Day 1 and Day 6 representing “old” solutions using BE-SPME by GC-FID and reported as (μmol as 2,3-dimethylnaphthalene/mL PDMS). Nominal loading rates were used in statistical endpoint evaluations.
New solution pH measurements did not vary by more than 0.57 units from the old test solution. Dissolved oxygen (DO) remained at or above 6.50 mg/L throughout the test. Temperatures in the exposure chambers ranged from 24.0 - 26.0 ºC.
No observation of test substance insolubility (surface slicks, precipitates, and adherence to the test chamber) was noted during the time of organism observations. No abnormal behavior or appearance was observed in the control or lowest five treatment groups. No observations of aborted eggs or immobilized neonates were noted in the control or treatment groups throughout the exposure. Immobilized test organisms were observed on Day 1 in 0.80, 2.0 and 5.0 mg/L treatment groups. There was 10% immobilization in the 0.80 mg/L group by Day 1, 20% immobilization in the 2.0 mg/L group by Day 2 and 100% immobilization in the 5mg/L group by Day 3.
Both acute (LL50) and chronic (EL10, EL20, NOEL, LOEL) endpoints were calculated for this study. The acute endpoint was calculated using 48h immobilization data and the chronic endpoints were calculated using reproduction data gathered throughout the duration of the study.
PROTOCOL DEVIATION
The protocol required that the pH should not vary by more than 0.3 units between corresponding
new and old solutions however the pH varied from 0.34-0.57 units on several occasions.
No critical effects were observed in the control group and the validity of the study was maintained. - Results with reference substance (positive control):
- No chronic reference toxicity test was performed at the testing facility.
- Reported statistics and error estimates:
- The EL10 and EL20 chronic endpoints were determined based on neonate counts and nominal loading rates relative to the control value. The EL10 and EL20 values with associated confidence intervals were calculated based on a non-linear regression model, the Hill Model, using the Benchmark Dose method (USEPA, 2010).
The NOEL and LOEL chronic endpoint data was determined based on nominal loading rates and reproduction data using JMP v. 14.1 (JMP, 2018). The Shapiro-Wilk goodness of fit test was used to determine if the data was normally distributed (Shapiro and Wilk, 1965). An ANOVA (Analysis of Variance) was used to determine homogeneity of variance of reproduction data. Nonparametric comparison with control using the Steel method (Steel, 1959) was performed to determine the NOEL and LOEL of nominal loading rates vs. total number of neonates produced in three broods. This statistical analysis identified the 0.02 mg/L level as statistically significant, but due to no effect at the consecutive three higher levels, it may be interpreted that it is not justifiable as a LOEL. - Validity criteria fulfilled:
- yes
- Conclusions:
- Chronic (EL10, EL20, NOEL, LOEL) endpoints were calculated for this study. The chronic endpoints were calculated using reproduction data gathered throughout the 6 days of the study and were based on nominal rates:
EL10: 0.49 (97.5% confidence interval of 0.28-0.71) mg/L
EL20: 0.72 (97.5% confidence interval of 0.49 -0.94) mg/L
NOEL: 0.32 mg/L
LOEL: 0.80 mg/L - Executive summary:
This study was conducted for the Sponsor to evaluate the effects of the water accommodated fractions (WAFs) of Naphtha (petroleum), steam-cracked middle arom (CAS: 68477-54-3)on survival and reproductive output of the cladoceran, Ceriodaphnia dubia, in a 3-brood semi-static dose-response test.
WAFs were prepared daily for each treatment group by adding the appropriate amount of the test substance using gas tight (glass and stainless steel) syringes to moderately hard reconstituted water with micronutrients in glass aspirator bottles sealed with PTFE screw caps. The WAFs were held in an environmental chamber with a 16:8 light cycle at approximately test temperature, and were mixed with Teflon-coated stir bars on magnetic stir plates for approximately 24 ± 1h. The control WAF was prepared in the same manner without test substance addition. At the end of mixing, the WAFs were allowed to settle for approximately 1h ± 30 mins. At the end of each settling period, approximately 100 mL of the WAF solution was removed and discarded, then solutions were removed from the mixing vessels through the outlet at the bottom of the vessels and placed into the respective test chambers. All test chambers contained no headspace. The WAF loading rates tested were control (0), 0.02, 0.05, 0.13, 0.32, 0.80, 2.0, and 5 mg/L.
Analytical verification of the test solutions 0.05, 0.32. 0.80 and 5 mg/L and the control were performed on Day 0 and Day 5 “new” solutions and Day 1 and Day 6 “old” solutions using the automated biomimetic extraction technique employing solid phase micro-extraction (BE-SPME). Nominal loading rates were used in statistical endpoint evaluations.
Chronic (EL10, EL20, NOEL, LOEL) endpoints were calculated for this study. The chronic endpoints were calculated using reproduction data gathered throughout the 6 days of the study and were based on nominal rates:
EL10: 0.49 (97.5% confidence interval of 0.28-0.71) mg/L
EL20: 0.72 (97.5% confidence interval of 0.49 -0.94) mg/L
NOEL: 0.32 mg/L
LOEL: 0.80 mg/L
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
- Justification for type of information:
- See Attached Justification
- Qualifier:
- according to guideline
- Guideline:
- other: REACH Guidance on QSARs R.6
- Principles of method if other than guideline:
- - Software tool(s) used including version:
See Field 'Attached justification'
- Model(s) used: See Field 'Attached justification'
- Model description: See Field 'Attached justification'
- Justification of QSAR prediction: see field 'Attached justification' - GLP compliance:
- not specified
- Specific details on test material used for the study:
- HC Block Type Carbon Number Concentration (%w/w)
Mono-Aromatic (MoAr) 7 0.75
8 35.64
9 17.80
10 2.99
12 0.35
13 0.23
Di-Aromatic (DiAr) 10 0.22
n-Cyclohexane (n-CC6) 7 0.11
iso-Naphthenic (i-N) 8 0.10
n-Olefin 5 0.58
6 0.17
9 0.61
10 23.76
i-Olefin 6 0.21
n-Paraffin (n-P) 7 0.12
8 0.18
9 0.30
Total 84.13
The constituents that could fit in more than one HC block, or that did not fit in any specific block, were placed in a relevant block that presented the highest toxicity. - Analytical monitoring:
- not required
- Test organisms (species):
- Daphnia magna
- Test type:
- other: QSAR Calculation
- Total exposure duration:
- 21 d
- Reference substance (positive control):
- not required
- Key result
- Duration:
- 21 d
- Dose descriptor:
- EL10
- Effect conc.:
- 0.493 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- other: QSAR prediction
- Basis for effect:
- reproduction
- Remarks on result:
- other: QSAR predicted value
- Validity criteria fulfilled:
- yes
- Conclusions:
- The QSAR predicted 21-d EL10 (reproduction) for D. magna was 0.493 mg/L.
- Executive summary:
The QSAR predicted 21 -d EL10 (reproduction) of the UVCB substance for D. magna was 0.493 mg/L. The model used for the calculations was the PETROTOX v4.0 model. PETROTOX is a well-documented and peer reviewed model that is widely used across the chemicals industry. This substance fits within the criteria of the model and there are no reservations about the validity of the model runs. It is expected that this data is reliable with restrictions.
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
- Justification for type of information:
- See Attached Justification
- Qualifier:
- according to guideline
- Guideline:
- other: REACH Guidance on QSARs R.6
- Principles of method if other than guideline:
- - Software tool(s) used including version:
See Field 'Attached justification'
- Model(s) used: See Field 'Attached justification'
- Model description: See Field 'Attached justification'
- Justification of QSAR prediction: see field 'Attached justification' - GLP compliance:
- not specified
- Specific details on test material used for the study:
- HC Block Type Carbon Number Concentration (%w/w)
Mono-Aromatic (MoAr) 7 2.49
8 4.24
9 66.33
10 5.12
Naphthenic Mono-Aromatic (NMAr) 9 19.58
Total 97.76
The constituents that could fit in more than one HC block, or that did not fit in any specific block, were placed in a relevant block that presented the highest toxicity. - Analytical monitoring:
- not required
- Test organisms (species):
- Daphnia magna
- Test type:
- other: QSAR Calculation
- Total exposure duration:
- 21 d
- Reference substance (positive control):
- not required
- Key result
- Duration:
- 21 d
- Dose descriptor:
- EL10
- Effect conc.:
- 1.36 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- other: QSAR prediction
- Basis for effect:
- reproduction
- Remarks on result:
- other: QSAR predicted value
- Validity criteria fulfilled:
- yes
- Conclusions:
- The QSAR predicted 21-d EL10 (reproduction) for D. magna was 1.36 mg/L.
- Executive summary:
The QSAR predicted 21 -d EL10 (reproduction) of the UVCB substance for D. magna was 1.36 mg/L. The model used for the calculations was the PETROTOX v4.0 model. PETROTOX is a well-documented and peer reviewed model that is widely used across the chemicals industry. This substance fits within the criteria of the model and there are no reservations about the validity of the model runs. It is expected that this data is reliable with restrictions.
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
- Justification for type of information:
- See Attached Justification
- Qualifier:
- according to guideline
- Guideline:
- other: REACH Guidance on QSARs R.6
- Principles of method if other than guideline:
- - Software tool(s) used including version:
See Field 'Attached justification'
- Model(s) used: See Field 'Attached justification'
- Model description: See Field 'Attached justification'
- Justification of QSAR prediction: see field 'Attached justification' - GLP compliance:
- not specified
- Specific details on test material used for the study:
- HC Block Type Carbon Number Concentration (%w/w)
n-olefin 5 0.57
9 1.24
10 76.50
11 14.84
12 1.21
Total 94.37
The constituents that could fit in more than one HC block, or that did not fit in any specific block, were placed in a relevant block that presented the highest toxicity. - Analytical monitoring:
- not required
- Test organisms (species):
- Daphnia magna
- Test type:
- other: QSAR Calculation
- Total exposure duration:
- 21 d
- Reference substance (positive control):
- not required
- Key result
- Duration:
- 21 d
- Dose descriptor:
- EL10
- Effect conc.:
- 0.219 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- other: QSAR prediction
- Basis for effect:
- reproduction
- Remarks on result:
- other: QSAR predicted value
- Validity criteria fulfilled:
- yes
- Conclusions:
- The QSAR predicted 21-d EL10 (reproduction) for D. magna was 0.219 mg/L.
- Executive summary:
The QSAR predicted 21 -d EL10 (reproduction) of the UVCB substance for D. magna was 0.219 mg/L. The model used for the calculations was the PETROTOX v4.0 model. PETROTOX is a well-documented and peer reviewed model that is widely used across the chemicals industry. This substance fits within the criteria of the model and there are no reservations about the validity of the model runs. It is expected that this data is reliable with restrictions.
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
- Justification for type of information:
- See Attached Justification
- Qualifier:
- according to guideline
- Guideline:
- other: REACH Guidance on QSARs R.6
- Principles of method if other than guideline:
- - Software tool(s) used including version:
See Field 'Attached justification'
- Model(s) used: See Field 'Attached justification'
- Model description: See Field 'Attached justification'
- Justification of QSAR prediction: see field 'Attached justification' - GLP compliance:
- not specified
- Specific details on test material used for the study:
- HC Block Type Carbon Number Concentration (%w/w)
Mono-Aromatic (MoAr) 7 1.55
8 31.91
9 17.86
10 2.89
12 2.29
13 0.67
Di-Aromatic (DiAr) 10 1.03
13 0.28
14 0.24
15 0.28
n-Olefin 5 0.27
10 21.06
11 0.41
i-Olefin 9 0.30
Total 81.03
The constituents that could fit in more than one HC block, or that did not fit in any specific block, were placed in a relevant block that presented the highest toxicity. - Analytical monitoring:
- not required
- Test organisms (species):
- Daphnia magna
- Test type:
- other: QSAR Calculation
- Total exposure duration:
- 21 d
- Reference substance (positive control):
- not required
- Key result
- Duration:
- 21 d
- Dose descriptor:
- EL10
- Effect conc.:
- 0.347 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- other: QSAR prediction
- Basis for effect:
- reproduction
- Remarks on result:
- other: QSAR predicted value
- Validity criteria fulfilled:
- yes
- Conclusions:
- The QSAR predicted 21-d EL10 (reproduction) for D. magna was 0.347 mg/L.
- Executive summary:
The QSAR predicted 21 -d EL10 (reproduction) of the UVCB substance for D. magna was 0.347 mg/L. The model used for the calculations was the PETROTOX v4.0 model. PETROTOX is a well-documented and peer reviewed model that is widely used across the chemicals industry. This substance fits within the criteria of the model and there are no reservations about the validity of the model runs. It is expected that this data is reliable with restrictions.
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
- Justification for type of information:
- See Attached Justification
- Qualifier:
- according to guideline
- Guideline:
- other: REACH Guidance on QSARs R.6
- Principles of method if other than guideline:
- - Software tool(s) used including version:
See Field 'Attached justification'
- Model(s) used: See Field 'Attached justification'
- Model description: See Field 'Attached justification'
- Justification of QSAR prediction: see field 'Attached justification' - GLP compliance:
- not specified
- Specific details on test material used for the study:
- HC Block Type Carbon Number Concentration (%w/w)
Mono-Aromatic (MoAr) 8 1.51
9 42.57
10 7.04
Naphthenic Mono-Aromatic (NMAr) 9 0.98
10 0.36
Di-Aromatic (DiAr) 10 8.24
11 0.85
n-Olefin 5 0.22
9 0.13
10 26.65
11 4.67
Total 93.21
The constituents that could fit in more than one HC block, or that did not fit in any specific block, were placed in a relevant block that presented the highest toxicity. - Analytical monitoring:
- not required
- Test organisms (species):
- Daphnia magna
- Test type:
- other: QSAR Calculation
- Total exposure duration:
- 21 d
- Reference substance (positive control):
- not required
- Key result
- Duration:
- 21 d
- Dose descriptor:
- EL10
- Effect conc.:
- 0.447 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- other: QSAR prediction
- Basis for effect:
- reproduction
- Remarks on result:
- other: QSAR predicted value
- Validity criteria fulfilled:
- yes
- Conclusions:
- The QSAR predicted 21-d EL10 (reproduction) for D. magna was 0.447 mg/L.
- Executive summary:
The QSAR predicted 21 -d EL10 (reproduction) of the UVCB substance for D. magna was 0.447 mg/L. The model used for the calculations was the PETROTOX v4.0 model. PETROTOX is a well-documented and peer reviewed model that is widely used across the chemicals industry. This substance fits within the criteria of the model and there are no reservations about the validity of the model runs. It is expected that this data is reliable with restrictions.
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
- Justification for type of information:
- See Attached Justification
- Qualifier:
- according to guideline
- Guideline:
- other: REACH Guidance on QSARs R.6
- Principles of method if other than guideline:
- - Software tool(s) used including version:
See Field 'Attached justification'
- Model(s) used: See Field 'Attached justification'
- Model description: See Field 'Attached justification'
- Justification of QSAR prediction: see field 'Attached justification' - GLP compliance:
- not specified
- Specific details on test material used for the study:
- HC Block Type Carbon Number Concentration (%w/w)
Mono-Aromatic (MoAr) 6 0.57
7 0.19
9 0.50
12 0.98
13 0.31
Di-Aromatic (DiAr) 10 0.27
n-Olefin 5 0.63
10 62.38
11 23.68
Total 89.52
The constituents that could fit in more than one HC block, or that did not fit in any specific block, were placed in a relevant block that presented the highest toxicity. - Analytical monitoring:
- not required
- Test organisms (species):
- Daphnia magna
- Test type:
- other: QSAR Calculation
- Total exposure duration:
- 21 d
- Reference substance (positive control):
- not required
- Key result
- Duration:
- 21 d
- Dose descriptor:
- EL10
- Effect conc.:
- 0.218 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- other: QSAR prediction
- Basis for effect:
- reproduction
- Remarks on result:
- other: QSAR predicted value
- Validity criteria fulfilled:
- yes
- Conclusions:
- The QSAR predicted 21-d EL10 (reproduction) for D. magna was 0.218 mg/L.
- Executive summary:
The QSAR predicted 21 -d EL10 (reproduction) of the UVCB substance for D. magna was 0.218 mg/L. The model used for the calculations was the PETROTOX v4.0 model. PETROTOX is a well-documented and peer reviewed model that is widely used across the chemicals industry. This substance fits within the criteria of the model and there are no reservations about the validity of the model runs. It is expected that this data is reliable with restrictions.
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
- Justification for type of information:
- See Attached Justification
- Qualifier:
- according to guideline
- Guideline:
- other: REACH Guidance on QSARs R.6
- Principles of method if other than guideline:
- - Software tool(s) used including version:
See Field 'Attached justification'
- Model(s) used: See Field 'Attached justification'
- Model description: See Field 'Attached justification'
- Justification of QSAR prediction: see field 'Attached justification' - GLP compliance:
- not specified
- Specific details on test material used for the study:
- HC Block Type Carbon Number Concentration (%w/w)
Mono-Aromatic (MoAr) 6 0.44
7 2.99
8 5.10
9 83.11
10 0.95
Naphthenic Mono-Aromatic (NMAr) 9 2.40
Di-Aromatic (DiAr) 10 1.13
n-Olefin 10 0.54
n-Paraffin (n-P) 10 0.14
11 0.33
Total 97.12
The constituents that could fit in more than one HC block, or that did not fit in any specific block, were placed in a relevant block that presented the highest toxicity. - Analytical monitoring:
- not required
- Test organisms (species):
- Daphnia magna
- Test type:
- other: QSAR Calculation
- Total exposure duration:
- 21 d
- Reference substance (positive control):
- not required
- Key result
- Duration:
- 21 d
- Dose descriptor:
- EL10
- Effect conc.:
- 1.27 mg/L
- Nominal / measured:
- estimated
- Conc. based on:
- other: QSAR prediction
- Basis for effect:
- reproduction
- Remarks on result:
- other: QSAR predicted value
- Validity criteria fulfilled:
- yes
- Conclusions:
- The QSAR predicted 21-d EL10 (reproduction) for D. magna was 1.27 mg/L.
- Executive summary:
The QSAR predicted 21 -d EL10 (reproduction) of the UVCB substance for Daphnia magna was 0.1.27 mg/L. The model used for the calculations was the PETROTOX v4.0 model. PETROTOX is a well-documented and peer reviewed model that is widely used across the chemicals industry. This substance fits within the criteria of the model and there are no reservations about the validity of the model runs. It is expected that this data is reliable with restrictions.
Referenceopen allclose all
Results table:
Response variable |
Endpoints (based on nominal loading rates (mg/L)) |
||
Endpoint |
Mean |
Confidence interval results |
|
6 day reproduction |
EL10 |
0.85 |
0.50 -1.201 |
EL20 |
0.89 |
0.70 -1.071 |
|
NOEL |
0.41 |
|
|
LOEL |
0.91 |
|
1Values are 97.5% confidence intervals
2Values are 99% confidence intervals
Response Variable |
Endpoints |
|
Based on Nominal Loading Rates (mg/L) |
||
Reproduction |
EL10 |
0.81 (0.52–1.09) |
Values in parentheses ( ) are 95% confidence intervals.
Results table:
Response Variable |
Endpoints |
|
Based on Nominal Loading Rates (mg/L) |
||
Reproduction |
EL10 |
0.57 (0.42-0.72) |
Values in parentheses ( ) are 95% confidence intervals.
Results table
Response variable |
Endpoints (based on nominal loading rates (mg/L)) |
||
Endpoint |
Mean |
Confidence interval results |
|
6 day reproduction |
EL10 |
0.49 |
0.28-0.712 |
EL20 |
0.72 |
0.49-0.942 |
|
NOEL |
0.32 |
|
|
LOEL |
0.80 |
|
1Values are 95% confidence intervals
2Values are 97.5% confidence intervals
Description of key information
There are chronic daphnia studies available for 4 streams in this category. These were all performed in accordance with US EPA 821-R-02-13 using Ceriodaphnia dubia and exposures via WAFs. These studies reported an EL10 for reproduction ranging from 0.49 to 0.85 mg/L.
Key value for chemical safety assessment
Additional information
Experimental data:
EC 270-790-1 / CAS 68478-10-4 (EMBSI, 2020): This is a GLP compliant, guideline study considered adequate for assessment. The experimental EL10 value for Ceriodaphnia dubia 6 day reproduction was 0.85 mg/L.
EC 271-138-9 / CAS 68516-20-1 (EMBSI, 2020): This is a GLP compliant, guideline study considered adequate for assessment. The experimental EL10 value for Ceriodaphnia dubia 6 day reproduction was 0.49 mg/L.
EC 266-013-0 / CAS 65996-79-4 (EMBSI, 2019): This is a GLP compliant, guideline study considered adequate for assessment. The experimental EL10 value for Ceriodaphnia dubia 7 day reproduction was 0.81 mg/L.
EC 305-586-4 / CAS 94733-07-0 (EMBSI, 2019): This is a GLP compliant, guideline study considered adequate for assessment. The experimental EL10 value for Ceriodaphnia dubia 7 day reproduction was 0.57 mg/L.
QSAR predictions:
Species | Substance CAS No. | EL10 (mg/L) |
Daphnia magna | 68516-20-1 | 0.493 |
Daphnia magna | 101316-62-5 | 1.36 |
Daphnia magna | 68526-56-7 | 0.219 |
Daphnia magna | 68477-54-3 | 0.347 |
Daphnia magna | 94733-07-0 | 0.447 |
Daphnia magna | 68478-10-4 | 0.218 |
Daphnia magna | 65996-79-4 | 1.27 |
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.
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