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EC number: 237-695-7 | CAS number: 13927-71-4
- 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

Long-term toxicity to fish
Administrative data
Link to relevant study record(s)
- Endpoint:
- fish early-life stage toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- Analytical Sampling
Water samples were collected from one test chamber of each treatment and control group four, three, two and one day prior to test initiation to confirm concentrations after conditioning the diluter system for three, four, five and six days, respectively. Test solution samples were collected from one replicate test chamber in each treatment and control group at the beginning of the test, at approximately weekly intervals during the test and at the end of the test to measure concentrations of the test substance. Additional samples were collected on Days 14, 15, 21, 22, 28, 29 and 32 at the Study Director’s request. Sampling alternated between the replicate test chambers in each group at each sampling interval. Test solution samples (10.0 mL) were collected from mid-depth of the test solutions, were placed in glass scintillation vials and were processed immediately for LSC analysis or were held refrigerated for possible future analysis.
Additional samples were collected from the 0.13 and 2.0 µg/L treatment groups at the beginning of the test, on Day 14 of the test and at the end of the test to measure concentrations of the non-radiolabelled test substance. Sampling alternated between the replicate test chambers in the 0.13 and 2.0 µg/L treatment groups at each sampling interval. Test solution samples (10.0 mL) were collected from mid-depth of the test solutions, were placed in glass scintillation vials containing 10.0 mL of methanol and were processed immediately for LC/MS/MS analysis. - Vehicle:
- yes
- Remarks:
- Methanol
- Details on test solutions:
- * Preparation of Test Concentrations
Stock solutions of 14C-CDBC were prepared in methanol at a nominal concentration of 50.0 mg/L weekly during the test. Before measurement, the radiolabeled test substance was removed from frozen storage and allowed to equilibrate to room temperature. A 100-mL primary stock solution was prepared by weighing out 0.00500 g of the radiolabeled test substance. The radiolabeled test substance was then transferred to a 100 mL glass volumetric flask and brought to volume with methanol. The primary stock solutions were sonicated and then mixed by inversion and ranged in appearance from clear and brown to clear and brownish-yellow.
Stock solutions of 12C-CDBC were also prepared weekly during the study by dissolving non radiolabelled CDBC in methanol at a nominal concentration of 1.0 mg/mL. A 20-mL primary stock solution was prepared by weighing out 0.0200 g of the non-radiolabelled test substance. The non-radiolabeled test substance was then rinsed into a 20 mL glass volumetric flask and brought to volume with methanol. The primary stock solutions were sonicated for approximately 15 minutes and then mixed by inversion. The primary stock solutions appeared clear and black.
Dispensing stock solutions at nominal concentrations of 0.0013, 0.0025, 0.0050, 0.010 and 0.020 mg/mL were prepared in methanol using calculated volumes of the radiolabelled and non-radiolabelled stock solutions to achieve nominal radioactivities of 1.92 x 105, 1.00 x 105, 5.00 x 104, 2.50 x 104 and 1.25 x 104 dpm/µg, respectively in the stock solutions. The dispensing stock solutions were prepared on Day -7, -1, 6, 13, 20 and 27. The five dispensing stock solutions were injected into the diluter mixing chambers (at a rate of 20.0 µL/minute) where they were mixed with well water (at a rate of 200 mL/minute) to achieve the targeted test concentrations. The solvent control was prepared by injecting methanol into the mixing chamber, using the same target rates as mentioned above, for the solvent control. The concentration of methanol in the solvent control and all 14C-CDBC treatment groups was 0.1 mL/L.
The concentrations of the test were selected in consultation with the Sponsor and were based on the solubility of the test substance in water. The highest concentration selected, 2 µg/L, was selected based on preliminary solubility work conducted at Eurofins-Easton in dilution water.
* Dilution Water
The water used for testing was freshwater obtained from a well approximately 40 meters deep located on the Eurofins-Easton site. The well water was passed through a sand filter to remove particles greater than approximately 25 ¿m and pumped into a 37,800 L storage tank where the water was aerated with spray nozzles. Prior to use, the water was filtered to 0.45 ¿m to remove fine particles and was passed through an ultraviolet (UV) sterilizer.
The well water is characterized as moderately-hard water. The specific conductance, hardness, alkalinity, pH and total organic carbon (TOC) of the well water during the approximate four-week period immediately preceding the test were measured. - Test organisms (species):
- Pimephales promelas
- Details on test organisms:
- * Test Organism
The fathead minnow, Pimephales promelas, was selected as the test species for this study. Fathead minnows are one of the preferred fish species to test the toxicity of test substances during the early life-stages of fish. This species was selected for use in the test based upon past use and ease of handling in the laboratory. Fathead minnow embryos used in the test were obtained from cultures maintained by Eurofins Easton. Identification of the species was verified by the supplier of the original culture. The brood fish cultured at Eurofins were acclimated for at least 14 days prior to the collection of embryos for the test. During the 7 days immediately preceding the test, the brood fish used for the test showed no signs of disease or stress and there was no mortality.
On the morning of study initiation, the embryos were removed from the spawning substrates and examined under a dissecting microscope to select healthy, viable specimens at approximately the same stage of development. Embryos collected for use in the test were from 22 individual spawns and were < 24 hours old when the test was initiated. To initiate the test, groups of 1 to 3 embryos were impartially distributed among incubation cups until each cup contained 20 embryos. One cup was placed in each treatment and control test chamber.
Newly-hatched larvae were fed live brine shrimp nauplii (Artemia sp.) three times per day during the first seven days of post-hatch. Thereafter, they were fed live brine shrimp nauplii three times per day on weekdays and at least two times per day on weekends. Brine shrimp nauplii were obtained by hatching cysts purchased from Brine Shrimp Direct of Ogden, Utah. The concentrations of selected pesticides and organic and inorganic constituents in the Artemia cysts are measured annually and the results from the most recent analysis are presented in Appendix 3. Fish were not fed for at least 24 hours prior to the termination of the test to allow for clearance of the digestive tracts before weight measurements were made. To ensure that the feeding rate per fish remained constant, rations were adjusted at least weekly to account for losses due to mortality.
Biomass loading at the end of the test, based on the mean wet weight of the negative control group, was 0.0238 g of fish per liter of test solution that passed through the test chamber during a 24-hour period. Instantaneous loading (the total wet weight of fish per liter of water in the tank) at the end of the test was 0.245 g fish/L. - Test type:
- flow-through
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 32 d
- Remarks on exposure duration:
- 32 Days (4-Day Hatch and 28-Day Post-Hatch)
- Hardness:
- 136 - 144 mg/L as CaCO3
- Test temperature:
- 24.7 - 25.0 °C
- pH:
- 7.9 - 8.1
- Dissolved oxygen:
- 7.5 - 8.2 mg/L
- Salinity:
- -
- Conductivity:
- -
- Nominal and measured concentrations:
- nominal concentrations: 0.13 µg/L, 0.25 µg/L, 0.50 µg/L, 1.0 µg/L, 2.0 µg/L (The highest concentration was selected based on preliminary solubility work n in dilution water)
measured concentrations: 0.14 µg/L, 0.23 µg/L, 0.44 µg/L, 1.0 µg/L, 1.6 µg/L - Details on test conditions:
- * Test Apparatus
A continuous-flow diluter was used to deliver each concentration of the test substance, a solvent (HPLC grade methanol) control, and a negative (dilution water) control. Syringe pumps (Harvard Apparatus, Holliston, Massachusetts) were used to deliver the five test substance stock solutions and HPLC-grade methanol for the solvent control into mixing chambers assigned to each treatment and the solvent control. The syringe pumps were calibrated prior to the test and verified at the end of the test. The stock solutions were diluted with well water in the mixing chambers in order to obtain the desired test concentrations. The flow of dilution water to the mixing chambers was controlled by rotameters, which were calibrated prior to test initiation and verified at approximately weekly intervals during the test. The flow of test water from each mixing chamber was split and allowed to flow into four replicate test chambers. The proportion of the test water that was split into each replicate was checked prior to the test and at approximately weekly intervals during the test to ensure that flow rates varied by no more than ±10% of the mean for the four replicates. The diluter flow rate was adjusted to provide approximately 10 volume additions of test water in each test chamber per day. The general operation of the diluter was checked visually at least two times per day during the test and at least once at the end of the test. Periodically during the test, all organisms were transferred to clean test chambers to prevent the buildup of bacterial/fungal growth.
The test was conducted in a temperature-controlled environmental chamber designed to maintain the target test temperature throughout the test period. The test chambers were 9-L glass aquaria filled with approximately 7 L of test solution. The depth of the test water in a representative test chamber was 16 cm. Test chambers were labeled with the study number, test concentration and replicate designation. Embryos were held in incubation cups constructed from glass cylinders approximately 50 mm in diameter with 425 µm nylon screen mesh attached to the bottom with silicone sealant. The cups were suspended in the water column of each test chamber and attached to a rocker arm. The reciprocating motion of the rocker arm (4 rpm) facilitated circulation of test water around the embryos during incubation.
* Environmental Conditions
Ambient laboratory light was used to illuminate the test systems. Fluorescent light bulbs that emit wavelengths similar to natural sunlight were controlled by an automatic timer to provide a photoperiod of 16 hours of light and 8 hours of darkness. A 30-minute transition period of low light intensity was provided when lights went on and off to avoid sudden changes in lighting. Light intensity was measured at the water surface of one representative test chamber at test initiation using a SPER Scientific Model 840006 light meter.
The target test temperature during the test was 25 ± 1°C. Temperature was measured in each test chamber at the beginning of the test, weekly during the test and at the end of the test using a digital thermometer. Water temperature also was monitored continuously during the test in one negative control test chamber using a validated environmental monitoring system (Pointview Real-time Data Reporting System). The system measurements were calibrated prior to exposure initiation and verified or recalibrated approximately weekly during the test with a digital thermometer.
Dissolved oxygen and pH were measured in alternating replicates of each treatment and control group at the beginning of the test, weekly during the test and at the end of the test. Measurements of dissolved oxygen were made using a Thermo Scientific Orion Star A213 Benchtop RDO/DO meter and pH was measured using a Thermo Scientific Orion DUAL STAR pH/ISE meter.
Hardness, alkalinity and specific conductance were measured in alternating replicates of the negative control (dilution water) and the highest concentration treatment group at the beginning of the test, weekly during the test and at the end of the test. Hardness and alkalinity were measured by titration based on procedures in Standard Methods for the Examination of Water and Wastewater. Specific conductance was measured using a Thermo Scientific Orion Star A122 Portable Conductivity meter. - Reference substance (positive control):
- no
- Key result
- Duration:
- 32 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1.6 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat. (total fraction)
- Basis for effect:
- mortality
- Key result
- Duration:
- 32 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1.6 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat. (total fraction)
- Basis for effect:
- time to hatch
- Key result
- Duration:
- 32 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1.6 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat. (total fraction)
- Basis for effect:
- weight
- Key result
- Duration:
- 32 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1.6 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat. (total fraction)
- Basis for effect:
- length
- Details on results:
- * RESULTS
Measurement of Test Concentrations
Nominal concentrations selected for use in the study were 0.13, 0.25, 0.50, 1.0 and 2.0 µg/L. The concentrations were selected in consultation with the Sponsor and were based on the solubility of the test substance in water. The highest concentration selected, 2 µg/L, was selected based on preliminary solubility work in dilution water.During the course of the test, the appearance of the test solutions at these nominal concentrations was observed in both the diluter mixing chambers, where test substance stocks and dilution water were combined prior to delivery to the test chambers, and in the test chambers. The test solutions in the test chambers appeared clear and colorless during the test, with no evidence of precipitation observed in any control or treatment solution. The test solutions in the 0.13, 0.25 and 0.50 µg/L mixing chambers appeared clear and colorless during the test, with no evidence of precipitation observed. At test initiation, the test solutions in the 1.0 and 2.0 µg/L mixing chambers were clear and colorless. However, the toxicant delivery lines in the 1.0 and 2.0 µg/L mixing chambers had brown material on them and the sides of the mixing chambers were brown where the delivery lines were touching. At test termination, the test solutions in the 1.0 and 2.0 µg/L treatment groups mixing chambers were clear and colorless but the ends of the toxicant delivery lines were stained brown.
The measured concentrations by LSC of samples collected to verify the diluter system prior to the test ranged from 54.0 to 122% of nominal concentrations. Samples of the test solution samples collected during the test had measured concentrations by LSC that ranged from 63.2 to 161% of nominal concentrations. Additional samples were collected from various treatment groups on Days 14, 15, 21, 22, 28, 29 and 32 when unusually high or low recoveries were measured. The mean from all analyzed samples from the respective treatment groups on those days were calculated for each interval and used to calculate the mean measured concentrations. When the measured concentrations by LSC of test solution samples collected during the test were averaged, the mean measured test concentrations were 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L, which represented 104, 93, 87, 100 and 82% of nominal concentrations, respectively. The results of the study were based on the mean measured concentrations by LSC.
Samples of the test solution samples from the 0.14 and 1.6 µg/L treatment groups collected during the test had measured concentrations by LC/MS/MS that ranged from
Physical and Chemical Measurements of Water
Water temperatures were within the 25 ± 1°C range established for the test. Dissolved oxygen concentrations remained = 91% of air saturation (7.5 mg/L). Measurements of pH ranged from 7.9 to 8.2 during the test. Measurements of specific conductance, hardness and alkalinity were comparable between the control and treatment group and did not appear to be influenced by CDBC concentration. Light intensity at test initiation was 602 lux at the surface of the water of one representative test chamber.
Time to Hatch
Fathead minnow embryos began hatching on Day 3 of the test. The majority of fathead minnow embryos in the control and treatment replicates hatched on Day 4 of the test. Hatching reached > 90% in the control groups on Day 4 of the test, at which time the larvae were released to their respective test chambers. The mean time to hatch in the negative and solvent control groups was 3.96 and 4.07 days, respectively. There were no statistically significant differences in mean time to hatch between the negative and solvent control groups (p > 0.05). Therefore, the control data were pooled for comparisons with the treatment groups. The mean time to hatch in the pooled control and the 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L treatment groups was study day 4.01, 4.01, 4.01, 4.08, 4.07 and 4.16, respectively. According to Dunnett’s test, there was a statistically significant increase in the 1.6 µg/L treatment groups in comparison to the pooled control group (p = 0.05). However, this increase was very slight (3.7%) in comparison to the pooled control and was not considered to be biologically meaningful. Therefore, the NOEC and LOEC for time to hatch were determined to be 1.6 µg/L and > 1.6 µg/L, respectively.
Hatching Success
Hatching success in the negative and solvent control groups was 97.5 and 96.3%, respectively. There were no statistically significant differences in hatching success between the negative and solvent control groups (p > 0.05). Therefore, the control data were pooled for comparisons with the treatment groups. Hatching success in the pooled control and the 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L treatment groups was 96.9, 98.8, 93.8, 96.3, 96.3 and 96.3, respectively. According to the Jonckheere Terpstra step-down trend test, a statistically significant decreasing trend was not evident in the hatching success data (p > 0.05). Consequently, the NOEC and LOEC for hatching success were determined to be 1.6 µg/L and > 1.6 µg/L, respectively. Since there was a less than a 10% reduction in hatching success between the treatment group means and the pooled control mean, the LC10 and LC20 values were empirically estimated to be greater than the highest treatment group.
Post-Hatch and Overall Survival
At test termination, post-hatch larval survival in the negative and solvent control groups was 92.4 and 97.4%, respectively. There were no statistically significant differences in post-hatch larval survival between the negative and solvent control groups (p > 0.05). Therefore, the control data were pooled for comparisons with the treatment groups. Post-hatch larval survival in the pooled control and the 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L treatment groups was 94.9, 92.3, 98.7, 97.4, 94.4 and 89.6%, respectively. Overall survival in the negative and solvent control groups was 90.0 and 93.8%, respectively. There were no statistically significant differences in overall survival between the negative and solvent control groups (p > 0.05). Therefore, the control data were pooled for comparisons with the treatment groups. Overall survival at test termination in the pooled control and the 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L treatment groups was 91.9, 91.3, 92.5, 93.8, 91.3 and 86.3%, respectively. According to the Jonckheere Terpstra step-down trend test, a statistically significant decreasing trend was not evident in either the post-hatch survival data or overall survival data (p > 0.05). Consequently, the NOEC and LOEC for both post-hatch survival and overall survival were determined to be 1.6 µg/L and > 1.6 µg/L, respectively. Since there was a less than a 10% reduction in post-hatch and overall survival between the treatment group means and the pooled control mean, the LC10 and LC20 values were empirically estimated to be greater than the highest treatment group.
Biological Observations
In general, the majority of the fish in the control groups and in all treatment groups appeared normal throughout the test. Observations of unusual behavior or appearance included appearing small, weak, discolored (pale), morphologically deformed (e.g. crooked spine, curled, protrusion on side of abdomen, and swollen abdomen) and lying on the bottom of the tank with little motion other than minor gill movement. However, these observations were infrequent, did not follow a dose-responsive pattern, and were comparable in the controls.
Total Length
Mean total length at test termination in the negative and solvent control groups was 22.6 and 22.4 mm, respectively. There were no statistically significant differences in mean total length between the negative and solvent control groups (p > 0.05). Therefore, the control data were pooled for comparisons with the treatment groups. Mean total length at test termination in the pooled control and the 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L treatment groups was 22.5, 22.1, 21.8, 22.0, 21.8 and 22.3 mm, respectively. According to Dunnett’s test, there were statistically significant decreases in the 0.14, 0.23, 0.44 and 1.0 µg/L treatment groups in comparison to the pooled control group (p = 0.05). However, the decreases in the 0.14, 0.23, 0.44 and 1.0 µg/L treatment groups were not considered to be treatment-related since the decrease was not dose responsive. Consequently, the NOEC and LOEC for total length were determined to be 1.6 µg/L and > 1.6 µg/L, respectively. Since there was a less than a 10% inhibition for total length between the treatment group means and the pooled control mean, the IC10 and IC20 values were empirically estimated to be greater than the highest treatment group.
Wet Weight
Mean wet weight at test termination in the negative and solvent control groups was 85.8 and 79.9 mg, respectively. There was a statistically significant difference between the negative and solvent control groups (p = 0.05), therefore, the treatment groups were compared to the solvent control group for analysis. Mean wet weight at test termination in the 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L treatment groups was 79.0, 77.2, 77.1, 72.2 and 79.9 mg, respectively. According to Dunnett’s test, there was a statistically significant decrease in the 1.0 µg/L treatment group in comparison to the solvent control group (p = 0.05). However, the decrease was not considered to be treatment-related since the decrease was not dose-responsive. Consequently, the NOEC and LOEC for wet weight were determined to be 1.6 µg/L and > 1.6 µg/L, respectively. Since there was a less than a 10% inhibition for wet weight between the treatment group means and the solvent control mean, the IC10 and IC20 values were empirically estimated to be greater than the highest treatment group.
Dry Weight
Mean dry weight at test termination in the negative and solvent control groups was 17.6 and 16.5 mg, respectively. There was a statistically significant difference between the negative and solvent control groups (p = 0.05), therefore, the treatment groups were compared to the solvent control group for analysis. Mean dry weight at test termination in the 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L treatment groups was 16.0, 16.2, 15.8, 15.0 and 16.1 mg, respectively. According to Dunnett’s test, there was a statistically significant decrease in the 1.0 µg/L treatment group in comparison to the solvent control group (p = 0.05). However, the decrease in the 1.0 µg/L treatment group was not considered to be treatment-related since the decrease was not dose responsive. Consequently, the NOEC and LOEC for dry weight were determined to be 1.6 µg/L and > 1.6 µg/L, respectively. Since there was a less than a 10% inhibition for dry weight between the treatment group means and the solvent control mean, the IC10 and IC20 values were empirically estimated to be greater than the highest treatment group. - Results with reference substance (positive control):
- -
- Reported statistics and error estimates:
- Test endpoints analyzed statistically included time to hatch, hatching success, post-hatch larval survival, overall survival and growth (total length, wet weight and dry weight). Data from the negative and solvent control groups for each parameter were compared using a t-test. Since no differences were detected between the two control groups (p > 0.05) for time to hatch, hatching success, post-hatch survival, overall survival and total length, the control data for these parameters were pooled for comparison among the treatment groups. There were statistically significant differences between the two control groups (p = 0.05) for wet weight and dry weight. Therefore, the treatment group data for dry weight were compared to the solvent control data.
The statistical tests were performed using a personal computer with SAS software.
The results of the statistical analyses were used to aid in the determination of the NOEC, LOEC and LC/ICx values. However, scientific judgement was used to determine if statistical differences were biologically meaningful, and if the data followed a concentration-dependent response. The LOEC was defined as the lowest tested concentration at which the test substance is observed to have had a statistically significant adverse effect (at p = 0.05) on time to hatch, hatching success, survival or growth when compared to the controls. However, all test concentrations above the LOEC should have a harmful effect equal to or greater than those observed at the LOEC. The NOEC was defined as the test concentration immediately below the LOEC, which when compared with the control groups, had no statistically significant adverse effect (at p = 0.05). - Validity criteria fulfilled:
- yes
- Conclusions:
- CONCLUSIONS
Fathead minnows (Pimephales promelas) were exposed to CDBC at mean measured concentrations of 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L under flow-through conditions for 32 days (a 4-day hatching period plus a 28-day post-hatch growth period). The highest concentration selected for the test was based on preliminary solubility work conducted in dilution water. There were no biologically meaningful, statistically significant treatment-related effects on time to hatch, hatching success, post-hatch larval survival, overall survival or growth at concentrations = 1.6 µg/L. Consequently, the NOEC and LOEC were 1.6 µg/L and > 1.6 µg/L, respectively.
Lethal concentrations (LC10 and LC20) for the hatching success, post-hatch larval survival and overall survival endpoints were empirically estimated to be greater than the highest test concentrations, since there was less than a 10% reduction in any treatment group when compared to the pooled control group. Inhibition concentrations (IC10 and IC20) for the total length, wet weight and dry weight endpoints could not be estimated, since there was a less than a 10% inhibition between the treatment group means and the pooled or solvent control.
In conclusion, no effect was measured up to the limit of solubility of the test item. - Executive summary:
The objective of this study, conducted according to the OECD guideline 210, was to determine the effects of copper bis(dibutyldithiocarbamate) (CDBC) on the time to hatch, hatching success, survival, and growth of fathead minnows,Pimephales promelas, during early life-stage development.
Fathead minnow embryos were exposed to a geometric series of five test concentrations, a negative (dilution water) control and a solvent control (0.1 mL/L HPLC-grade methanol) under flow-through conditions. The exposure period included a 4-day embryo hatching period, and a 28-day post-hatch juvenile growth period. Nominal test concentrations were 0.13, 0.25, 0.50, 1.0 and 2.0 µg/L. The concentrations were selected in consultation with the Sponsor and were based on the solubility of the test substance in water. The highest concentration selected, 2 µg/L, was selected based on preliminary solubility work conducted in dilution water.
There were no biologically meaningful, statistically significant treatment-related effects on time to hatch, hatching success, post-hatch larval survival, overall survival or growth at concentrations = 1.6 µg/L. Consequently, the NOEC and LOEC were 1.6 µg/L and > 1.6 µg/L, respectively.
Lethal concentrations (LC10 and LC20) for the hatching success, post-hatch larval survival and overall survival endpoints were empirically estimated to be greater than the highest test concentrations, since there was less than a 10% reduction in any treatment group when compared to the pooled control group. Inhibition concentrations (IC10 and IC20) for the total length, wet weight and dry weight endpoints could not be estimated, since there was a less than a 10% inhibition between the treatment group means and the pooled or solvent control.
LENGTH OF EXPOSURE: 32 Days (4-Day Hatch and 28-Day Post-Hatch)
TEST ORGANISMS: Fathead Minnow (Pimephales promelas)
AGE OF TEST ORGANISMS: Embryos (<24 hours old)
TEST CONCENTRATIONS: Nominal Mean Measured
Negative Control <LOQ
Solvent Control <LOQ
0.13 µg/L 0.14 µg/L
0.25 µg/L 0.23 µg/L
0.50 µg/L 0.44 µg/L
1.0 µg/L 1.0 µg/L
2.0 µg/L 1.6 µg/L
RESULTS: Based on mean measured test concentrations
EndpointLC10/IC10
(µg/L)
LC20/IC20
(µg/L)
NOEC
(µg/L)
LOEC
(µg/L)
Time to Hatch
--
--
1.6 µg/L
> 1.6 µg/L
Hatching Success
> 1.6 µg/L1
> 1.6 µg/L1
1.6 µg/L
> 1.6 µg/L
Post-Hatch Larval Survival
> 1.6 µg/L1
> 1.6 µg/L1
1.6 µg/L
> 1.6 µg/L
Overall Survival
> 1.6 µg/L1
> 1.6 µg/L1
1.6 µg/L
> 1.6 µg/L
Total Length
> 1.6 µg/L1
> 1.6 µg/L1
1.6 µg/L
> 1.6 µg/L
Wet Weight
> 1.6 µg/L1
> 1.6 µg/L1
1.6 µg/L
> 1.6 µg/L
Dry Weight
> 1.6 µg/L1
> 1.6 µg/L1
1.6 µg/L
> 1.6 µg/L
No-Observed-Effect Concentration:
1.6 µg/L
Lowest-Observed-Effect-Concentration:
> 1.6 µg/L
1 Empirically estimated to be greater than the highest test concentration, since there was less than a 10% decrease in any treatment group in comparison to the pooled or solvent control.
Reference
Measured Concentrations of14C-CDBC in the Primary Stock Solutions
Sample Type |
Sample Number (15175-) |
Nominal 14C-CDBC Concentration (mg/L) |
Specific Activity (dpm/mg) |
Total [14C] Found (dpm) |
14C-CDBC Equivalents Found1 (mg/L) |
Percent of Nominal |
Stock |
090419-1-A |
50.0 |
250072 |
586397 |
46.9 |
93.8 |
Stock |
090419-1-B |
50.0 |
250072 |
575431 |
46.0 |
92.0 |
Stock |
090419-1-C |
50.0 |
250072 |
580306 |
46.4 |
92.8 |
|
|
|
|
Mean |
46.4 |
92.9 |
|
|
|
|
|
|
|
Stock |
091119-2-A |
50.0 |
250072 |
617894 |
49.4 |
98.8 |
Stock |
091119-2-B |
50.0 |
250072 |
639329 |
51.1 |
102 |
Stock |
091119-2-C |
50.0 |
250072 |
623518 |
49.9 |
99.7 |
|
|
|
|
Mean |
50.1 |
100 |
|
|
|
|
|
|
|
Stock |
091819-3-A |
50.0 |
250072 |
572157 |
45.8 |
91.5 |
Stock |
091819-3-B |
50.0 |
250072 |
520621 |
41.6 |
83.3 |
Stock |
091819-3-C |
50.0 |
250072 |
589406 |
47.1 |
94.3 |
|
|
|
|
Mean |
44.8 |
89.7 |
|
|
|
|
|
|
|
Stock |
092619-5-A |
50.0 |
250072 |
585952 |
46.9 |
93.7 |
Stock |
092619-5-B |
50.0 |
250072 |
578658 |
46.3 |
92.6 |
Stock |
092619-5-C |
50.0 |
250072 |
581590 |
46.5 |
93.0 |
|
|
|
|
Mean |
46.6 |
93.1 |
|
|
|
|
|
|
|
Stock |
100219-6-A |
50.0 |
250072 |
579998 |
46.4 |
92.8 |
Stock |
100219-6-B |
50.0 |
250072 |
580930 |
46.5 |
92.9 |
Stock |
100219-6-C |
50.0 |
250072 |
588185 |
47.0 |
94.1 |
|
|
|
|
Mean |
46.6 |
93.3 |
|
|
|
|
|
|
|
Stock |
100919-7-A |
50.0 |
250072 |
673641 |
53.9 |
108 |
Stock |
100919-7-B |
50.0 |
250072 |
670814 |
53.6 |
107 |
Stock |
100919-7-C |
50.0 |
250072 |
673152 |
53.8 |
108 |
|
|
|
|
Mean |
53.8 |
108 |
1 14C-CDBC Equivalents = (Total [14C] found / Specific Activity/ Sample Volume (mL)) x Dilution Factor Sample Volume = 0.0500 mL; Dilution Factor = 1.00 |
Measured Concentrations of14C-CDBC in Dispensing Stock Solutions
Sample Type |
Sampling Interval (Day) |
Sample Number (524A-138-) |
Nominal CDBC Concentration (mg/mL) |
Specific Activity (dpm/mg) |
Total [14C] Found (dpm) |
14C-CDBCEquivalents[AG1] [YB2] Found1 (mg/mL) |
Percent Recovery |
Background |
-1 |
BKG |
-- |
-- |
0 |
-- |
-- |
Dosing Solution |
-1 |
S-1 |
0.0013 |
192350 |
49666 |
0.00129 |
99.3 |
Dosing Solution |
-1 |
S-2 |
0.0025 |
100011 |
50648 |
0.00253 |
101 |
Dosing Solution |
-1 |
S-3 |
0.0050 |
50003 |
50546 |
0.00505 |
101 |
Dosing Solution |
-1 |
S-4 |
0.010 |
25001 |
50955 |
0.0102 |
102 |
Dosing Solution |
-1 |
S-5 |
0.020 |
12500 |
51110 |
0.0204 |
102 |
1 14C-CDBC Equivalents = (Total [14C] found / Specific Activity / Sample Volume (mL)) x Dilution Factor Sample Volume = 0.200 mL; Dilution Factor = 1.00 |
Measured Concentrations of12C-CDBC in Dispensing Stock Solutions
Sample Number (524A-138-) |
Sampling Interval (Day) |
CDBC Nominal Concentration (mg/mL) |
12C-CDBC Nominal Concentration (mg/mL)1 |
12C-CDBC Measured Concentration (mg/mL)2 |
12C-CDBC Percent of Nominal1,3 |
S-1 |
-1 |
0.0013 |
0.00030 |
0.000471 |
157 |
S-2 |
-1 |
0.0025 |
0.0015 |
0.00147 |
98.0 |
S-3 |
-1 |
0.0050 |
0.0040 |
0.00410 |
103 |
S-4 |
-1 |
0.010 |
0.0090 |
0.00931 |
103 |
S-5 |
-1 |
0.020 |
0.019 |
0.0194 |
102 |
1 The stock solutions for the test system were prepared as a mixture of radiolabeled and non-labeled material. Nominal concentrations of CDBC in the stock solutions were corrected for the ratio of12C-CDBC in each test solution. The corrected nominal values were calculated as follows [Cold Material (mg)/(Hot Material (mg) + Cold Material (mg))] x CDBC Nominal Concentration. 2 Results were generated using Analyst Version 1.6.3. Manual calculations may differ slightly. 3 Calculated by Excel 2010, based upon measured values of12C-CDBC/12C-CDBC nominal concentration. |
Measured Concentrations of14C-CDBC in Pretest Verification Samples
|
Nominal CDBC Concentration (µg/L) |
Sample Number (524A-138-) |
Sampling Interval (Day) |
Specific Activity (dpm/µg) |
Total [14C] Found (dpm)1 |
14C-CDBC Equivalents Found1, 2 (µg/L) |
Percent of Nominal |
|
|
Negative Control |
PT-1-RE3 |
-4 |
NA |
< LOQ |
< LOQ |
-- |
|
|
(0.0) |
PT-11 |
-3 |
NA |
< LOQ |
< LOQ |
-- |
|
|
|
PT-21 |
-2 |
NA |
< LOQ |
< LOQ |
-- |
|
|
|
PT-31 |
-1 |
NA |
< LOQ |
< LOQ |
-- |
|
|
|
|
|
|
|
|
|
|
|
Solvent Control |
PT-2-RE3 |
-4 |
NA |
< LOQ |
< LOQ |
-- |
|
|
(0.0) |
PT-12 |
-3 |
NA |
< LOQ |
< LOQ |
-- |
|
|
|
PT-22 |
-2 |
NA |
< LOQ |
< LOQ |
-- |
|
|
|
PT-32 |
-1 |
NA |
< LOQ |
< LOQ |
-- |
|
|
|
|
|
|
|
|
|
|
|
0.13 |
PT-3-RE3 |
-4 |
192350 |
305 |
0.159 |
122 |
|
|
|
PT-13 |
-3 |
192350 |
221 |
0.115 |
88.4 |
|
|
|
PT-23 |
-2 |
192350 |
199 |
0.103 |
79.6 |
|
|
|
PT-33 |
-1 |
192350 |
226 |
0.117 |
90.4 |
|
|
|
|
|
|
|
|
|
|
|
0.25 |
PT-4-RE3 |
-4 |
100011 |
225 |
0.225 |
90.0 |
|
|
|
PT-14 |
-3 |
100011 |
210 |
0.210 |
84.0 |
|
|
|
PT-24 |
-2 |
100011 |
203 |
0.203 |
81.2 |
|
|
|
PT-34 |
-1 |
100011 |
225 |
0.225 |
90.0 |
|
|
|
|
|
|
|
|
|
|
|
0.50 |
PT-5-RE3 |
-4 |
50003 |
165 |
0.330 |
66.0 |
|
|
|
PT-15 |
-3 |
50003 |
210 |
0.420 |
84.0 |
|
|
|
PT-25 |
-2 |
50003 |
165 |
0.330 |
66.0 |
|
|
|
PT-35 |
-1 |
50003 |
209 |
0.418 |
83.6 |
|
|
|
|
|
|
|
|
|
|
|
1.0 |
PT-6-RE3 |
-4 |
25001 |
167 |
0.668 |
66.8 |
|
|
|
PT-16 |
-3 |
25001 |
176 |
0.704 |
70.4 |
|
|
|
PT-26 |
-2 |
25001 |
176 |
0.704 |
70.4 |
|
|
|
PT-36 |
-1 |
25001 |
201 |
0.804 |
80.4 |
|
|
|
|
|
|
|
|
|
|
|
2.0 |
PT-7-RE3 |
-4 |
12500 |
257 |
2.06 |
103 |
|
|
|
PT-17 |
-3 |
12500 |
202 |
1.62 |
80.8 |
|
|
|
PT-27 |
-2 |
12500 |
135 |
1.08 |
54.0 |
|
|
|
PT-37 |
-1 |
12500 |
201 |
1.61 |
80.4 |
|
1 The limit of quantification (LOQ) for14C-CDBC in water is 50 dpm, based upon the LOQ of the liquid scintillation counter (LSC). 2 14C-CDBC Equivalents = [(Total dpm found/Sample volume)/Specific activity] Sample volume = 0.0100 L 3 Sample was reanalyzed due to instrument contamination. The sample was analyzed on a different instrument and the result is presented here. |
|
Measured Concentrations of14C-CDBC in Test Solution Samples
|
Nominal CDBC Concentration (µg/L) |
Sample Number (524A-138-) |
Sampling Interval (Day) |
Specific Activity (dpm/µg) |
Total [14C] Found (dpm)1 |
14C-CDBC Equivalents Found1, 2, 3 (µg/L) |
Percent of Nominal |
Mean14C-CDBC Equivalents Found (µg/L)/Standard Deviation/ %CV1,2,3 |
Mean Percent of Nominal1 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Negative |
4 |
0 |
NA |
< LOQ |
< LOQ |
-- |
-- |
-- |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Control |
16 |
7 |
NA |
< LOQ |
< LOQ |
-- |
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
26 |
14 |
NA |
< LOQ |
< LOQ |
-- |
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
41 |
21 |
NA |
< LOQ |
< LOQ |
-- |
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
55 |
28 |
NA |
< LOQ |
< LOQ |
-- |
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
67 |
32 |
NA |
< LOQ |
< LOQ |
-- |
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
|
|
|
|
|
|
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Solvent |
5 |
0 |
NA |
< LOQ |
< LOQ |
-- |
-- |
-- |
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Control |
17 |
7 |
NA |
< LOQ |
< LOQ |
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27 |
14 |
NA |
< LOQ |
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42 |
21 |
NA |
< LOQ |
< LOQ |
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56 |
28 |
NA |
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68 |
32 |
NA |
< LOQ |
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0.13 |
6 |
0 |
192350 |
266 |
0.138 |
106 |
0.14 |
104 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
18 |
7 |
192350 |
277 |
0.144 |
111 |
SD = 0.0117 |
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
28 |
14 |
192350 |
248 |
0.129 |
99.2 |
%CV = 8.66 |
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
43 |
21 |
192350 |
257 |
0.134 |
103 |
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
57 |
28 |
192350 |
223 |
0.116 |
89.2 |
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
69 |
32 |
192350 |
286 |
0.149 |
114 |
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
Hatching Success of Fathead Minnow Embryos Exposed to CDBC
Mean Measured Concentration (µg/L) |
Rep |
Number of Embryos Exposed |
Cumulative No. of Dead Embryos |
Cumulative Number Hatched by Day of Test |
Replicate % Hatching Success |
Treatment % Hatching Success |
|||||
Day 1 |
Day 2 |
Day 3 |
Day 41 |
Day 5 |
|
||||||
0.44 |
A |
20 |
1 |
0 |
0 |
0 |
17 |
19 |
|
95 |
96.3 |
|
B |
20 |
0 |
0 |
0 |
0 |
19 |
20 |
|
100 |
|
|
C |
20 |
0 |
0 |
0 |
0 |
18 |
20 |
|
100 |
|
|
D |
20 |
2 |
0 |
0 |
0 |
17 |
18 |
|
90 |
|
|
|
|
|
|
|
|
|
|
|
|
|
1.0 |
A |
20 |
0 |
0 |
0 |
0 |
17 |
20 |
|
100 |
96.3 |
|
B |
20 |
0 |
0 |
0 |
0 |
20 |
20 |
|
100 |
|
|
C |
20 |
3 |
0 |
0 |
0 |
15 |
17 |
|
85 |
|
|
D |
20 |
0 |
0 |
0 |
0 |
20 |
20 |
|
100 |
|
|
|
|
|
|
|
|
|
|
|
|
|
1.6 |
A |
20 |
0 |
0 |
0 |
0 |
14 |
20 |
|
100 |
96.3 |
|
B |
20 |
0 |
0 |
0 |
1 |
16 |
20 |
|
100 |
|
|
C |
20 |
3 |
0 |
0 |
0 |
17 |
17 |
|
85 |
|
|
D |
20 |
0 |
0 |
0 |
0 |
16 |
20 |
|
100 |
|
1Day 4 of test = Day 0 post-hatch. Any embryos remaining after Day 4 of the test were kept in the egg cup until death occurred, or until hatching, at which time the newly hatched larva was added to the larval growth chamber. |
Survival to Day 28 Post-Hatch of Fathead Minnow Larvae Exposed toCDBC
Nominal Concentration (µg/L) |
Rep. |
Number of Larvae Hatched |
Live Counts by Day Post-Hatch1 |
Replicate Percent Post-Hatch Survival2 |
Treatment Percent Post-Hatch Survival2 |
Replicate Percent Overall Survival2 |
Treatment Percent Overall Survival2 |
||||||||
Day 0 |
Day 3 |
Day 7 |
Day 10 |
Day 14 |
Day 17 |
Day 21 |
Day 24 |
Day 28 |
|||||||
Negative Control |
A |
20 |
20 |
20 |
19 |
18 |
18 |
18 |
18 |
18 |
18 |
90.0 |
92.3 |
90.0 |
90.0 |
|
B |
18 |
173 |
18 |
17 |
17 |
17 |
17 |
17 |
17 |
17 |
94.4 |
|
85.0 |
|
|
C |
20 |
183 |
20 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
95.0 |
|
95.0 |
|
|
D |
20 |
20 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
90.0 |
|
90.0 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Solvent Control |
A |
19 |
143 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
94.7 |
97.4 |
90.0 |
93.8 |
|
B |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
100 |
|
90.0 |
|
|
C |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
100 |
|
100 |
|
|
D |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
19 |
19 |
19 |
95.0 |
|
95.0 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
0.14 |
A |
20 |
20 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
90.0 |
92.4 |
90.0 |
91.3 |
|
B |
19 |
19 |
19 |
18 |
16 |
16 |
16 |
16 |
16 |
16 |
84.2 |
|
80.0 |
|
|
C |
20 |
183 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
100 |
|
100 |
|
|
D |
20 |
20 |
20 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
95.0 |
|
95.0 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
0.23 |
A |
19 |
19 |
19 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
94.7 |
98.7 |
90.0 |
92.5 |
|
B |
17 |
17 |
17 |
17 |
17 |
17 |
17 |
17 |
17 |
17 |
100 |
|
85.0 |
|
|
C |
20 |
173 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
100 |
|
100 |
|
|
D |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
100 |
|
95.0 |
|
Survival to Day 28 Post-Hatch of Fathead Minnow Larvae Exposed toCDBC
Nominal Concentration (µg/L) |
Rep. |
Number of Larvae Hatched |
Live Counts by Day Post-Hatch1 |
Replicate Percent Post-Hatch Survival2 |
Treatment Percent Post-Hatch Survival2 |
Replicate Percent Overall Survival2 |
Treatment Percent Overall Survival2 |
||||||||
Day 0 |
Day 3 |
Day 7 |
Day 10 |
Day 14 |
Day 17 |
Day 21 |
Day 24 |
Day 28 |
|||||||
0.44 |
A |
19 |
173 |
19 |
19 |
19 |
19 |
19 |
19 |
18 |
18 |
94.7 |
97.4 |
90.0 |
93.8 |
|
B |
20 |
193 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
100 |
|
100 |
|
|
C |
20 |
183 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
95.0 |
|
95.0 |
|
|
D |
18 |
173 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
100 |
|
90.0 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
1.0 |
A |
20 |
173 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
100 |
94.8 |
100 |
91.3 |
|
B |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
100 |
|
100 |
|
|
C |
17 |
153 |
14 |
14 |
14 |
14 |
14 |
14 |
14 |
14 |
82.4 |
|
70.0 |
|
|
D |
20 |
20 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
95.0 |
|
95.0 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
1.6 |
A |
20 |
143 |
20 |
19 |
19 |
19 |
19 |
19 |
19 |
19 |
95.0 |
89.6 |
95.0 |
86.3 |
|
B |
20 |
163 |
18 |
17 |
17 |
17 |
17 |
17 |
17 |
17 |
85.0 |
|
85.0 |
|
|
C |
17 |
17 |
16 |
16 |
16 |
16 |
16 |
15 |
15 |
15 |
88.2 |
|
75.0 |
|
|
D |
20 |
163 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
18 |
90.0 |
|
90.0 |
|
1 Day 0 post-hatch = Day 4 of test. Live counts made on Days 0 through 28 post-hatch were actual counts made at transfer to clean test chambers and at test termination. 2 Replicate and treatment group means were calculated using Excel in full-precision. Manual calculations may differ slightly. 3 Additional larvae hatched on Day 1 post-hatch and were released into the larval growth chamber. |
Summary of Clinical Observations of Fathead Minnow Larvae Exposed toCDBCDuring the 28-Day Post-Hatch Period
Mean Measured Concentration (µg/L) |
Rep. |
Clinical Observations by Day Post-Hatch1 |
|
|||
Day 0 |
Day 3 |
Day 7 |
Day 10 |
|||
Negative Control |
A |
AN |
20 AN |
1 S,MC,R,D; 18 AN |
1 M; 18 AN |
|
B |
AN |
18 AN |
17 AN |
17 AN |
||
C |
AN |
20 AN |
1 M; 19 AN |
19 AN |
||
D |
AN |
1 X; 1 M; 18 AN |
18 AN |
18 AN |
||
Solvent Control |
A |
AN |
1 M; 18 AN |
18 AN |
18 AN |
|
B |
AN |
18 AN |
18 AN |
18 AN |
||
C |
AN |
20 AN |
1 S,W; 19 AN |
1 S,W; 19 AN |
||
D |
AN |
20 AN |
1 S; 19 AN |
1 S; 19 AN |
||
0.14 |
A |
1 CU,W; rest AN |
1 X; 1 M; 18 AN |
18 AN |
18 AN |
|
|
B |
1 W; rest AN |
19 AN |
1 X; 1 S,W; 17 AN |
2 M; 16 AN |
|
|
C |
AN |
20 AN |
20 AN |
20 AN |
|
|
D |
1 CU,R; rest AN |
1 CU,W; 19 AN |
19 AN |
19 AN |
|
|
||||||
0.23 |
A |
AN |
19 AN |
1 M; 18 AN |
18 AN |
|
|
B |
AN |
17 AN |
17 AN |
17 AN |
|
|
C |
AN |
20 AN |
20 AN |
20 AN |
|
|
D |
AN |
19 AN |
19 AN |
19 AN |
Summary of Clinical Observations of Fathead Minnow Larvae Exposed to CDBC During the 28-Day Post-Hatch Period
Mean Measured Concentration (µg/L) |
Rep. |
Clinical Observations by Day Post-Hatch1 |
|
||||
Day 0 |
Day 3 |
Day 7 |
Day 10 |
||||
0.44 |
A |
AN |
19 AN |
19 AN |
19 AN |
||
B |
AN |
20 AN |
20 AN |
20 AN |
|||
C |
AN |
1 M; 19 AN |
19 AN |
19 AN |
|||
D |
AN |
18 AN |
1 S; 17 AN |
1 S; 17 AN |
|||
1.0 |
A |
AN |
20 AN |
20 AN |
20 AN |
||
|
B |
AN |
20 AN |
20 AN |
20 AN |
||
|
C |
AN |
3 M; 14 AN |
14 AN |
14 AN |
||
|
D |
AN |
1 M; 19 AN |
19 AN |
19 AN |
||
|
|||||||
1.6 |
A |
AN |
20 AN |
1 M; 19 AN |
19 AN |
||
|
B |
1 W; rest AN |
2 M; 18 AN |
1 M; 17 AN |
17 AN |
||
|
C |
1 CU,R; rest AN |
1 M; 16 AN |
16 AN |
16 AN |
||
|
D |
AN |
1 M; 18 AN |
18 AN |
18 AN |
||
|
Summary of Clinical Observations of Fathead Minnow Larvae Exposed to CDBC During the 28-Day Post-Hatch Period
Mean Measured Concentration (µg/L) |
Rep. |
Clinical Observations by Day Post-Hatch1 |
|
||||
Day 14 |
Day 17 |
Day 21 |
Day 24 |
Day 28 |
|||
Negative Control |
A |
18 AN |
18 AN |
18 AN |
18 AN |
18 AN |
|
B |
17 AN |
17 AN |
17 AN |
17 AN |
17 AN |
||
C |
19 AN |
19 AN |
19 AN |
19 AN |
19 AN |
||
D |
18 AN |
18 AN |
18 AN |
18 AN |
18 AN |
||
Solvent Control |
A |
18 AN |
18 AN |
18 AN |
18 AN |
18 AN |
|
B |
18 AN |
18 AN |
18 AN |
18 AN |
18 AN |
||
C |
1 S; 19 AN |
1 S,P; 19 AN |
1 S,P; 19 AN |
1 SA; 19 AN |
1 S,MC,SA; 19 AN |
||
D |
1 S; 19 AN |
1 S,W; 19 AN |
1 M; 19 AN |
19 AN |
19 AN |
||
0.14 |
A |
18 AN |
1 S; 17 AN |
1 S; 17 AN |
1 S; 17 AN |
1 S; 17 AN |
|
|
B |
16 AN |
2 S; 14 AN |
1 S; 15 AN |
16 AN |
2 S; 14 AN |
|
|
C |
20 AN |
1 MC; 19 AN |
1 MC; 19 AN |
20 AN |
1 S; 19 AN |
|
|
D |
19 AN |
19 AN |
19 AN |
19 AN |
19 AN |
|
|
|||||||
0.23 |
A |
18 AN |
18 AN |
18 AN |
18 AN |
18 AN |
|
|
B |
17 AN |
17 AN |
17 AN |
17 AN |
17 AN |
|
|
C |
20 AN |
20 AN |
20 AN |
20 AN |
20 AN |
|
|
D |
19 AN |
19 AN |
19 AN |
19 AN |
19 AN |
Summary of Clinical Observations of Fathead Minnow Larvae Exposed to CDBC During the 28-Day Post-Hatch Period
Mean Measured Concentration (µg/L) |
Rep. |
Clinical Observations by Day Post-Hatch1 |
|
|||||
Day 14 |
Day 17 |
Day 21 |
Day 24 |
Day 28 |
||||
0.44 |
A |
19 AN |
19 AN |
19 AN |
18 AN |
18 AN |
||
B |
20 AN |
20 AN |
20 AN |
20 AN |
20 AN |
|||
C |
19 AN |
19 AN |
19 AN |
19 AN |
19 AN |
|||
D |
1 S; 17 AN |
1 S; 17 AN |
1 S; 17 AN |
1 S; 17 AN |
1 S; 17 AN |
|||
1.0 |
A |
20 AN |
20 AN |
20 AN |
20 AN |
20 AN |
||
|
B |
20 AN |
20 AN |
20 AN |
20 AN |
20 AN |
||
|
C |
14 AN |
14 AN |
14 AN |
14 AN |
14 AN |
||
|
D |
19 AN |
19 AN |
19 AN |
19 AN |
19 AN |
||
|
||||||||
1.6 |
A |
19 AN |
19 AN |
19 AN |
19 AN |
19 AN |
||
|
B |
17 AN |
17 AN |
17 AN |
17 AN |
17 AN |
||
|
C |
16 AN |
16 AN |
1 X; 15 AN |
15 AN |
15 AN |
||
|
D |
18 AN |
18 AN |
18 AN |
18 AN |
18 AN |
||
1 Day 0 post-hatch = Day 4 of test. Observations made on Days 3 through 28 post-hatch were made at transfer to clean test chambers and at test termination. Observations: AN = appear normal; S = small; X = dead; P = protrusion on left side of abdomen; W = weak; M = missing and assumed dead; SA = swollen abdomen; MC= crooked spine; CU = curled; R = lying on bottom with little motion other than gill or appendage movement; D = discoloration (pale). |
|
Description of key information
Fathead minnows (Pimephales promelas) were exposed to CDBC at mean measured concentrations of 0.14, 0.23, 0.44, 1.0 and 1.6 µg/L under flow-through conditions for 32 days (a 4-day hatching period plus a 28-day post-hatch growth period). The highest concentration selected for the test was based on preliminary solubility work conducted in dilution water. There were no biologically meaningful, statistically significant treatment-related effects on time to hatch, hatching success, post-hatch larval survival, overall survival or growth at concentrations = 1.6 µg/L. Consequently, the NOEC and LOEC were 1.6 µg/L and > 1.6 µg/L, respectively.
In conclusion, no effect was measured up to the limit of solubility of the test item.
Key value for chemical safety assessment
Fresh water fish
Fresh water fish
- Effect concentration:
- 1.6 µg/L
Additional information
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.
