4,4'-bis(chloromethyl)-1,1'-biphenyl

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 June 2016 to 01 July 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF 2-7-2-1 The guidelines related to the study reports for the registration application of pesticide (Ref. No. 12-Nousan-8147 on 24 November 2000) & Ref. No.13-Seisan-3986 on 10 October 2001.

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Guidelines for studies on the new chemical substance as required by the Law Concerning the Evaluation of Chemical Substances and Regulation of their Manufacture, etc. (Chemical Substance Control Law) 1973.

Version / remarks:

amended 2009 under the reference of YAKUSHOKHATSU No. 1121002, SEIKYOKU No.2 and KANPOKIHATSU No. 021121002 and partially amended 2006 as the joint ordinance of The Japanese Ministry of Economy Trade and Industry (METI), Ministry of Health, Labour and Welfare (MHLW) and Ministry of the Environment (MOE).

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source: provided by sponsor
- Lot/batch No.of test material: 150701

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Controlled room temperature (15-25 ºC, below 70 RH%), protected from humidity, under inert gas.

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

TEST MATERIAL FORMULATION
The test material was poorly soluble in water therfore a test solution was prepared using a saturated solution method. A saturated Test material stock solution (water) was prepared by dispersing/dissolving the amount of test material into the test medium (ISO Medium) one day before the treatment. This solution was shaken for about 24 hours at approximately 30 °C and then was equilibrated for about 6 hours at approximately 20 °C. The non-dissolved test material was removed by filtration through a fine (0.22 µm) filter to give the 100 % saturated solution. As Limit Test was carried out, further dilution of stock solution was not be performed.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Justification for species: Daphnia magna is the standard species of the acute immobilisation test.
- Age of animals: They were less than 24 h old at the beginning of the test.
- Feeding during test: The animals were not fed during the test.
- Food type: The animals were not fed during the test.
- Number of animals: There were 20 animals in the test group and control gorup respectively, divided into 4 replicates.
- Breeding: The health of stock animals is continuously monitored by visual daily checking. Abnormal behaviour or significant decrease of population is recorded.

ACCLIMATION
- Acclimation period: There was no acclimatization because the water used was similar to the culture water.
- Type and amount of food: The animals were not fed during the test.

Test type:

semi-static

Water media type:

other: dilution water (ISO-medium)

Limit test:

yes

Total exposure duration:

48 h

Hardness:

The reconstituted water (ISO medium) had a total hardness of 249 mg/L (as CaCO3).

Test temperature:

The water temperature was measured at the start and at the end of each renewal period in each test vessel. The test temperature was in the range of 20.1 – 20.3 °C measured in the test vessels. The additionally measured temperature in the climate chamber was between 19.7 and 20.7 °C.

pH:

The pH of the test solution was not adjusted and not varied by more than 1.5 units in any one test. The pH was measured at the start and at the end of each renewal period in each test vessel and was in the range of 7.45 – 8.07.

Dissolved oxygen:

The dissolved oxygen concentration was measured in each test vessel at the start and at the end of each renewal period and was in the range of 7.4 – 8.5 mg/L.

Salinity:

no data

Conductivity:

no data

Nominal and measured concentrations:

Only one test concentration at the solubility level of the test material in the test medium (100 % saturated solution) and one control group were tested in a Limit Test under semi-static conditions.
Nominal loading rate: 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beaker
- Material, size, headspace, fill volume: At least 5 mL test solution/animal
- Renewal rate of test solution (frequency/flow rate): The frequency of the water renewal periods were 24 hours.
- No. of organisms per vessel: Five animals per vessel
- No. of vessels per concentration (replicates): 1 replicates per concentration
- No. of vessels per control (replicates): 1 replicates per concentration

OTHER TEST CONDITIONS
- Adjustment of pH: No adjustment of pH was carried out.
- Photoperiod: The test was carried out in 16-hour light and 8-hour dark cycle.

EFFECT PARAMETERS MEASURED/ OBSERVATIONS:
- The immobility or mortality of the Daphnia was determined by visual observation 24 and 48 hours after the start of the test. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker are considered to be immobile.
- The number of immobilised animals and the percentage of immobility were determined at 24 and 48 hours.
- The water temperature, oxygen concentrations and pH of the control and the test solution were measured at the beginning and at the end of the renewal periods.


RANGE-FINDING STUDY
- Test concentrations: Nominal concentrations 0.1, 1.0, 10 and 100 mg/L
- No. of organisms per vessel: Five animals per vessel.
- No. of vessels per concentration (replicates): 2 replicates per concentration.
- No. of vessels per control (replicates): 2 replicates per concentration.
- Results used to determine the conditions for the definitive study: The choice of the test concentration was done on the basis of the results of a preliminary range-finding test. Because significantly toxic response was not observed during the preliminary concentration range-finding test, only one test concentration at the solubility level of the test material in the test medium (100 % saturated solution) and one control group were tested in a limit test under semi-static conditions.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 other: % saturated solution

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 other: % saturated solution

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 other: % saturated solution

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 other: % saturated solution

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 other: % saturated solution

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Immobilisation
The number of immobilised animals and the percentage of immobility were determined at the 24th and 48th hour. No animals treated with the test substance were immobile at the 24 h and 48 h time points.
In addition to immobility, no abnormal behaviour or appearance of test animals was detected.

Results with reference substance (positive control):

For the evaluation of the quality of the Daphnia clone and the experimental conditions, Potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.
The 24h EC50 of the positive control was 1.16 mg/L (95 % confidence limits: 1.07 – 1.25 mg/L).

Reported statistics and error estimates:

No statistical analysis was performed because of the lack of toxic effects. The EC50, NOEC, LOEC and EC100 values were determined directly from the raw data.

VALIDITY

There were no immobilized animals in the control group and the dissolved oxygen concentration at the end of the test in control and test vessels was more than 3 mg/L. All validity criteria were within acceptable limits and therefore the study can be considered as valid.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this Daphnia magna acute immobilisation study, the 24h and 48h EC50 value is >100 % saturated solution (nominal), the 48h NOEC is 100 % saturated solution (nominal), the 48h LOEC is >100 % saturated solution (nominal) and the 48h EC100 value is >100 % saturated solution (nominal).

Executive summary:

The acute toxicity of the test material to Daphnia magna was investigated in a GLP study which was conducted in accordance with standardised guidelines OECD 202, EU Method C.2 and the JMAFF guidelines under static conditions.

Acute toxicity of the test material on Daphnia magna was assessed in an Acute immobilisation test, over an exposure period of 48 hours in a semi-static test system. Toxic effects were not observed during the preliminary concentration range-finding test therefore a limit test was performed in the main experiment using only one test concentration at the limit of solubility of the test item in the test medium (100 % saturated solution) and one control group. The experiment was carried out without analytical support based on agreement with the Sponsor. Twenty animals, divided into four groups (glass beaker) of five animals each were used at the test concentration and for the control. No statistical analysis was performed because of the lack of toxic effects. The EC50, NOEC, LOEC and EC100 values were determined directly from the raw data. All validity criteria were met during this study therefore the study was valid.

Under the conditions of this Daphnia magna acute immobilisation study, the 24h and 48h EC50 value is >100 % saturated solution (nominal), the 48h NOEC is 100 % saturated solution (nominal), the 48h LOEC is >100 % saturated solution (nominal) and the 48h EC100 value is >100 % saturated solution (nominal).

Based on the results of this study, the test material had no toxic effect at saturation; the EC50 and the LOEC are higher than the solubility level of the test item in the test medium.

Description of key information

Under the conditions of this Daphnia magna acute immobilisation study, the 24h and 48h EC50 value is >100 % saturated solution (nominal), the 48h NOEC is 100 % saturated solution (nominal), the 48h LOEC is >100 % saturated solution (nominal) and the 48h EC100 value is >100 % saturated solution (nominal)

Additional information

The acute toxicity of the test material to Daphnia magna was investigated in a GLP study which was conducted in accordance with standardised guidelines OECD 202, EU Method C.2 and the JMAFF guidelines under static conditions. The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

Acute toxicity of 4,4’-bis(chloromethyl)-1,1’-biphenyl on Daphnia magna was assessed in an Acute immobilisation test, over an exposure period of 48 hours in a semi-static test system. Toxic effects were not observed during the preliminary concentration range-finding test therefore a limit test was performed in the main experiment using only one test concentration at the limit of solubility of the test item in the test medium (100 % saturated solution) and one control group. The experiment was carried out without analytical support based on agreement with the Sponsor. Twenty animals, divided into four groups (glass beaker) of five animals each were used at the test concentration and for the control. No statistical analysis was performed because of the lack of toxic effects. The EC50, NOEC, LOEC and EC100 values were determined directly from the raw data. All validity criteria were met during this study therefore the study was valid. Under the conditions of this Daphnia magna acute immobilisation study, the 24h and 48h EC50 value is >100 % saturated solution (nominal), the 48h NOEC is 100 % saturated solution (nominal), the 48h LOEC is >100 % saturated solution (nominal) and the 48h EC100 value is >100 % saturated solution (nominal).