Source: https://www.echa.europa.eu/web/guest/registration-dossier/-/registered-dossier/25698/6/4/2
Timestamp: 2020-05-28 01:30:17
Document Index: 1086068

Matched Legal Cases: ['art.\n14', 'art: 6', 'art: 5', 'art: 5', 'art: 6', 'art: 5', 'art: 5', 'art: 5', 'art: 49', 'art: 49', 'art: 51', 'art: 54', 'art: 50', 'art: 53', 'art: 48', 'art: 21']

05 Dec 2014 - 19 Jun 2015
1) The temperature dropped below 18 °C and was above 22 °C for few and short time periods (14.6 - 23.5 °C). 2) In all treatments tested the soil moisture was more than 10% higher at the end of the test than the initial moisture (26.6% to 41.0%).
see deviations for OECD 222
Concentration was only measured in the stock solution before application. HPLC-UV analysis of the test substance in stock solution confirmed the nominal concentration (nominal: 3.56 mg/mL; measured: 3.70 mg/mL).
- Method of mixing into soil: The test item was dissolved in an amount of acetone sufficient to prepare a stock solution. This stock solution was used to produce the various dosage solutions of the test item. An appropriate amount of the stock and the dosage solutions respectively was used to soak a portion of the quartz sand of the artificial soil by the dosage solution. The solvent was allowed to evaporate at room temperature for 1 h and 15 min under a fume hood. The test item coated fraction of the quartz sand was then incorporated into the remainder of the artificial soil by thoroughly mixing it in for approx. 10 min. Finally, the contaminated artificial soil was filled into the test vessels.
- Source: originally obtained in 1994 from Co. Landenberger, Schömberg, Germany
- Age at test initiation: adults with clitellum; ; at least 2 months old but not older than 1 year (age did not differ by more than 4 weeks)
- Weight at test initiation (mean and range, SD): 250 - 600 mg freshweight
- Feeding during test: Fed with finely ground cow manure (free of pharmaceuticals, growth promotors, provided by Jesus Bruderschaft e.V. Gnadenthal, Hünfelden, Germany). Food was first provided one day after test start. Afterwards, worms were fed every week during the first 4 weeks of exposure. 5 g was moistened with 10 mL deionised water and spread on the soil surface. After removing the adults after 28 d a further 5 g of food was mixed with artificial soil.
- Acclimation period: Earthworms were acclimatised in artificial soil under test conditions for at least 24 h before test start.
14.6 - 23.5 °C (mean 19.9 °C)
Control - Start: 6.0, End: 6.8
Solvent control - Start: 5.9, End: 6.9
5 mg/kg dw - Start: 5.9, End: 6.8
10 mg/kg dw - Start: 6.0, End: 6.8
20 mg/kg dw - Start: 5.9, End: 6.8
40 mg/kg dw - Start: 5.8, End: 6.9
80 mg/kg dw - Start: 5.8, End: 6.8
Control - Start: 49.0% of WHCmax, End: 69.1% of WHCmax
Solvent control - Start: 49.2% of WHCmax, End: 67.7% of WHCmax
5 mg/kg dw - Start: 51.3% of WHCmax, End: 65.6% of WHCmax
10 mg/kg dw - Start: 54.3% of WHCmax, End: 74.1% of WHCmax
20 mg/kg dw - Start: 50.4% of WHCmax, End: 69.1% of WHCmax
40 mg/kg dw - Start: 53.8% of WHCmax, End: 68.1% of WHCmax
80 mg/kg dw - Start: 48.8% of WHCmax, End: 64.7% of WHCmax
- Test container (material, size): Bellaplast containers (Company Kastelplast, Mainz, Germany) consisting of inert (non-toxic) plastic (Polystyrol). The test vessels have a base area of 11 x 15.5 cm (=170.5 cm²) and a height of 6 cm. The test vessels were covered with a transparent and perforated lid.
- Amount of soil or substrate: 0.5 kg/container
- No. of organisms per container (treatment): 10 per container
- Composition (if artificial substrate): 5% sphagnum peat (5% instead of 10% peat was used considering the influence on bioavailability), 20% kaolin clay (kaolinite content > 30%), ~74% quartz sand (fine sand content with particles between 50 and 200 microns higher than 50%), 0.3 - 1% Calcium carbonate (CaCO3). The pH value was adjusted to 6 ± 0.5 using calcium carbonate. The moisture content was adjusted to 40-60% of WHCmax using deionised water.
- Photoperiod: 16 h light/8 h darkness
- Light intensity: 414-580 lux
- Mortality after 28 d
- Biomass after 0 and 28 d
- Reproduction after 56 d
- Range finding study: yes (no details reported)
Nominal: Water control, solvent control and Nominal concentrations 5, 10, 20, 40 and 80 mg/kg soil dw
>= 80 mg/kg soil dw
> 80 mg/kg soil dw
40.9 mg/kg soil dw
- Mortality at end of exposure period: 2.5% at 10, 40 and 80 mg/kg soil dw (after 28 d)
- Changes in body weight of live adults (% of initial weight) at end of exposure period: see "Any other informations on results incl. tables"
- No. of offspring produced: see "Any other informations on results incl. tables"
- Morphological abnormalities: No other effects on behavior or morphology of the adult earthworms were observed.
- Behavioural abnormalities: No other effects on behavior or morphology of the adult earthworms were observed.
- Other biological observations: Almost the whole food was consumed by the earthworms in the test vessels of the control and at the concentrations of 5, 10, 20 and 40 mg test item/kg soil (dw) after 7 and 14 d. After 21 and 28 d of the application the food consumption was reduced compared to the time points before. At the concentration of 80 mg/kg soil dw half or less than half of the food was consumed after 7, 14, 21 and 28 d.
- Relevant effect levels: 2.5% mortality was observed at the control, no mortality at the concentrations of 178, 267, 400 and 900 mg boric acid/kg soil (dw) and 12.5% mortality at a concentration of 600 mg boric acid/kg soil (dw). Concerning reproduction the statistical analysis (Dunnett’s t-test) showed a significant difference between the control and all concentrations of the reference item tested. The EC50 value was calculated by Probit analysis using Linear Max. Likelihood Regression as 259.3 mg boric Acid/kg artificial soil (dw) (95% confidence limits = 235.8 - 284.4 mg boric Acid/kg artificial soil (dw)). The estimated EC50 value in this study (259.3 mg boric acid /kg soil (dw)) is slightly lower than recommended by the guideline. However, since the calculated EC50 is lower by factor of 1.5 than the recommended value of 400 mg boric acid/ kg soil (dw) acceptable sensitivity of the test system is assumed.
Biomass and reproduction data were checked for normality by R/s test procedure and for homogeneity by Cochran’s test. Solvent control and water control data were compared by the pairwise Student-t test (p ≤ 0.05, two-sided). Concerning biomass and reproduction no statistically significant difference between the solvent control and the water control was determined. Therefore, for further statistical calculations of the biomass and the reproduction data, the water control data have been excluded. Treatment means were compared by ANOVA followed by Dunnett’s test (p ≤ 0.05, two-sided for biomass one-sided for reproduction) and tested for statistically significant differences compared to the solvent control (Sachs 1982). The EC50 was determined using Probit analysis (Finney, 1971). The statistical software package ToxRat Professional 2.10 was used for these calculations.
Table 1: Effect on mortality (28 d), biomass (28 d) and reproduction (56 d)
Concentration [mg/kg soil dw]
Mean change of initial weight per replicate [%]
* Statistically significant different to solvent control (Dunnett’s test, 2-sided, p ≤0.05)
** Statistically significant different to solvent control (Dunnett’s test, 1-sided, p ≤0.05)
0 - 2.5% mortality of adult earthworms was observed within the treatments after 28 d of exposure. Mortality of 2.5% was observed at nominal concentrations of 20, 40 and 80 mg/kg dw, respectively. No mortality was recorded at 5 and 20 mg/kg dw.
Biomass increase was between 43.1 - 55.8% at test concentrations of 5 - 40 mg/kg dw after 28 d. Statistical analysis showed a significant difference (only 2.8% increase; Dunnett’s test, 2-sided; p ≤ 0.05) concerning biomass development of individual adults over 28 d between the solvent control and the highest concentration of the test item tested (80 mg/kg soil dw).
Statistical analysis (Dunnett’s test; 1-sided, p ≤ 0.05) showed a significant difference concerning the number of juveniles between the solvent control and the concentrations of 20, 40 and 80 mg/kg dw (79.9, 61.1, 1.5% juveniles compared to the solvent control, respectively). All other concentrations tested did not differ statistically significant from the control.
Almost the whole food was consumed by the earthworms in the test vessels of the control and at the concentrations of 5, 10, 20 and 40 mg test item/kg soil (dw) after 7 and 14 d. After 21 and 28 d of the application the food consumption was reduced compared to the time points before. At the concentration of 80 mg/kg soil dw half or less than half of the food was consumed after 7, 14, 21 and 28 d.
Behavior/Morphology
No other effects on behavior or morphology of the adult earthworms were observed.
Each replicate (containing 10 adults) to have produced ≥ 30 juveniles by the end of the test.
The coefficient of variation of reproduction is ≤ 30%.
Adult mortality over the initial 4 weeks of the test is ≤ 10%.
NOEC (56 d): 10 mg/kg soil dw based on reproduction
The present study was performed in accordance with OECD 222 and GLP. Adult earthworms (Eisenia fetida) were exposed to the test substance (5, 10, 20, 40 and 80 mg/kg dw) in artificial soil for a total of 56 d and biomass, reproduction and mortality were assessed. Mortality was assessed additionally after 14 d. The test substance concentrations were prepared using acetone as solvent. However, the solvent was completely evaporated before test initiation. With respect to the properties of the test substance, 5% instead of 10% peat was used. No mortality was observed in the control and in the solvent control and at 0 to 2.5% in all concentrations of the test item tested. Therefore, no statistical analysis was performed and thus the NOEC (14 d) was considered to be ≥ 80 mg/kg soil dw. The NOEC (28 d) based on biomass was 40 mg/kg soil dw whereas the NOEC (56 d) for reproduction was 10 mg/kg soil dw.
06 Dec 2006 - 21 Dec 2006
- Method of mixing into soil (if used): A stock solution was prepared by dissolving 3900 mg of 4-Chloro-3-methylphenol in 20 mL acetone. A sequential 1 :2 dilution series was prepared, starting with the 195.0 mg/mL stock solution by adding 7 mL of acetone to 7 mL of the. stock solution or dilution. Then 5 mL of the stock solution or corresponding dilution were added to 20 g fine quartz sand. The treated saod was left for at least one hour in a fume hood until the solvent had evaporated and was then mixed with a spoon. The artificial soil was then added to test item blended sand.
- Controls: yes, blank control
- Source: Bred by IBACON in a breeding medium of cattle manure, peat, sand and straw, mainly fed with cattle manure, stored at room temperature.
- Age at test initiation (mean and range, SD): 11 to 12 months, with clitellum
- Acclimation period: 1 day, in artificial soil
Initial pH: 5.6 - 5.7
End of the study: 5.3 - 5.5
- Test container (material, size): Normal glass bottling jars (1 L)
- Amount of soil or substrate: Approximately 500 g (dry weight equivalent) artificial soil.
- Soil texture: artificial soil according to guideline
- % sand: pproximately 74.8% fine quartz-sand (F34) containing more than 50% by mass of particle size 0.05 mm to 0.2 mm; (Quarzwerke Frechen, Frechen, Germany)
- % chalk: approximately 0.2% chalk (CaCO3) added to adjust pH to 6.0 ± 0.5 (Merck, Darmstadt, Germany)
- % clay: 20% Kaolin clay (Erbslöh, Lohrheim, Germany)
- % peat: 5.0% Sphagnum-peat, air-dried and finely ground (2 mm) (Floragard, Vertriebs GmbH für Gartenbau, Oldenburg, Germany)
- water content: Water content was checked at the start and termination of the experiment in one test unit per treatment group, according to DIN 19683. At experimental start: 21.3 % to 23.0%, corresponding to 53.3% to 57.5% of the max. warer holding capacity. At experimental end: 20.7% to 22.1 % corresponding to 51.8 % to 55.3 % of the max. water holding capacity.
- Light intensity: 410 - 750 Lux
- Mortality was recorded after 7 and 14 d after application
- Behavioural abnormalities was recorded after 7 and 14 d
- Mean body weight was determined at day 0 and 14 d
VEHICLE CONTROL PERFORMED: no (solvent was evaporated before application)
- Test concentrations: control, 1, 10, 100, 500 and 1000 mg/kg artificial soil (dry weight)
- Results used to determine the conditions for the definitive study: 100 % mortality at a concentration of 500 mg/kg
nominal: control and 15, 30, 60, 120, 240 and 480 mg/kg dw soil
139.4 mg/kg soil dw
- Mortality at end of exposure period: After 14 days of exposure no mortality was observed up to and including the concentration of 60.0 mg test item/kg soil. At the concentration of 120 mg test item/kg soil mortality increased to 17.5%.
- Changes in body weight of live adults (% of initial weight) at end of exposure period: The body weight changes of the earthworms were not significantly different compared to the control up to and including the concentration of 60.0 mg test item/kg soil. At the concentration of 120 mg test item/kg soil a statistically significant decrease of body weight was observed.
- Behavioural abnormalities: Worms were fidgety wriggling about at the concentration of 120 mg/kg soil after 7 days of exposure. This effect was not observed after 14 days of exposure. No additional behavioural effects were observed in any other concentration.
- Relevant effect levels: LC50 after 14 days was determined to be 27.0 mg test item/kg soil dry weight (95% confidence limits of 22.6 and 31.1)
Fisher extract test (mortality), Dunnett test (weight change), Probit Analysis (determination of LC50). The software used to perform the statistical analysis was ToxRatPro, version 2.09 (2005).
Table 1: Results obtained in the study with p-chloro-m-cresol after 7 and 14 days
[mg a.i./kg d. wt.s.]
Mortality 7 days [%]
Mortality 14 days [%]
n.s. not significantly different compared to the control, Fischer exact test, α = 0.05
*significantly different compared to the control, Fischer exact test, α = 0.05
Table 2: Earthworm body weight changes
Mean [mg/worm]
Difference %1)
1) % mean of 4 replicates
n.s. not significantly different compared to the control, Dunnett test, α = 0.05
*significantly different compared to the control, Dunnett test, α = 0.05
Mortality of control animals
LC50 (14 d) = 139.4 mg/kg dw
NOEC (14 d) = 60 mg/kg dw
The effect of p-chloro-m-cresol on earthworms was studied on Eisenia fetida (Lührs, 2007). The test was carried out according to OECD Guideline 207 (1984). The test item was mixed with artificial soil and adult earthworms were exposed for 14 days to the test vessels filled with artificial soil of the nominal concentrations of 15.0, 30.0, 60.0, 120 and 480 mg test item/kg dry weight soil. An acetone treated control was run in parallel. Morphological and behavioural changes and the number of surviving worms in each test vessel were recorded after 7 and 14 days. The LC50 value was calculated by probit analysis as 139.4 mg test item/kg dw. The NOECmortality as well as the NOECbiomass was determined to be 60 mg test item/kg dw. Worms were fidgety wriggling about at the concentration of 120 mg/kg soil after 7 days of exposure. This effect was not observed after 14 days of exposure. No additional behavioural effects were observed in any other concentration. The test was performed according to guideline and is considered acceptable since validity criteria are fulfilled (control mortality < 10%).
other: source, CAS 59-50-7, key rel 1, Scheffczyk 2015
other: Biomasss
source, CAS 59-50-7, key rel 1, Scheffczyk 2015
other: source, CAS 59-50-7, rel 1, Lührs 2007
NOEC (56 d): 10 mg/kg soil dw based on reproduction, read-across
No study investigating the toxicity of sodium p-chloro-m-cresolate (CAS 15733-22-9) to soil macroorganisms is available. Therefore, in accordance to Regulation (EC) No. 1907/2006 Annex XI, 1.5 a read-across to the source substance (p-chloro-m-cresol, CAS 59-50-7) which is the main transformation product of the target substance sodium p-chloro-m-cresolate (CAS 15733-22-9) is applied.
One key (Scheffczyk 2015) and one supporting study (Lührs 2007) are available investigating the toxicity of the source substance p-chloro-m-cresol (CAS 59-50-7) to earthworms. The key study was performed in accordance with OECD 222 and GLP. Adult earthworms (Eisenia fetida) were exposed to the test substance (5, 10, 20, 40 and 80 mg/kg dw) in artificial soil for a total of 56 d and biomass, reproduction and mortality were assessed. Mortality was assessed additionally after 14 d. The test substance concentrations were prepared using acetone as solvent. However, the solvent was completely evaporated before test initiation. With respect to the properties of the test substance, 5% instead of 10% peat was used. No mortality was observed in the control and in the solvent control and at 0 to 2.5% in all concentrations of the test item tested. Therefore, no statistical analysis was performed and thus the NOEC (14 d) was considered to be ≥ 80 mg/kg soil dw. The NOEC (28 d) based on biomass was 40 mg/kg soil dw whereas the NOEC (56 d) for reproduction was 10 mg/kg soil dw.
The supporting study was carried out according to OECD Guideline 207. The test item was mixed with artificial soil and adult earthworms were exposed for 14 days to the test vessels filled with artificial soil of the nominal concentrations of 15.0, 30.0, 60.0, 120 and 480 mg/kg dry weight soil. An acetone treated control was run in parallel. Morphological and behavioral changes and the number of surviving worms in each test vessel were recorded after 7 and 14 days. The LC50 value was calculated by probit analysis as 139.4 mg/kg soil dw. The NOECMortality as well as the NOECbiomass was determined to be 60 mg/kg soil dw.