4,4'-isopropylidenedicyclohexanol

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:

19 March 2007 to 11 April 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study undertaken by Japanese accredited laboratory to internationally recognised guidelines.

Qualifier:

according to guideline

Guideline:

other: Heisei 15/11/21-No.1121002 of the Pharmaceutical and Food Safety Bureau

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Heisei 15/11/13-No.2 of the Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Heisei No.031121002 of the Environmental Policy Bureau, Ministry of the Environment, Japan.

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Measurement of concentration of the test substance
Concentrations of the test substance were analyzed by using high-performance liquid chromatograph-mass spectrometer. For all concentration groups, total four times, i.e. at the start of the exposure, just before and after of water change and at the end of the exposure, concentrations of the test substance were analyzed. Note that at the start of the exposure and after water change, from the test vessel in which the test solution for 4 replicates were prepared at the same time, 100 ml of the test solution were taken and applied as the test solution for analysis. Further, before water change and at the end of the exposure, from the test vessel of 4 replicates for each concentration group, even amount of the test solution were taken and mixed to 100 ml of the test solution for the test solution for analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The test substance was added to the water for the test solution. Through 48 hours mixing by magnetic stirrer, in the dark place, at 20 °C ± 1 °C, original solution of the test substance (100 mg/L) was prepared. However, there were small amount of undissolved test substance in the original solution of the test substance. These were removed by filtration with glass fiber filter (GF/F [Whatman International Ltd.]). The test solutions for each concentration groups were prepared by adding the original solution of the test substance to the test water. Note that the test water at the time of preparation of the test solutions was used at 20 °C ± 1 °C by adjusting beforehand.
Note that, since the purity (95.9 %) was not considered for amount of the test substance, the nominal concentrations for each group were indicated in the concentration of the supplied substance. Further, the original solution of the test substance was prepared at use.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: National Institute for Environmental Studies
- Age at study initiation: < 24h
- Feeding during test: no

Test type:

semi-static

Water media type:

other: Dechlorinated tap water

Limit test:

no

Total exposure duration:

48 h

Hardness:

71 mg CaCO3/ L

Test temperature:

19.6 - 20.9 °C

pH:

7.9 - 8.1 (pH conditioning was not applied)

Dissolved oxygen:

7.4 - 9.0 mg/L

Nominal and measured concentrations:

Nominal: 6.5, 10, 16, 25, 40, 65 and 100 mg/L
Measured: 6.2, 9.3, 15.0, 24.6, 39.2, 63.0 and 97.7 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass beaker
- Type: loosely covered to reduce the loss of water due to evaporation and to avoid the entry of dust into the solutions.
- Material: glass
- Aeration: none
- Type of flow-through: semi-static
- Renewal rate of test solution: Every 24 hours
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod: 16 hours light, 8 hours dark

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.6
- Range finding study: yes
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 31 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 95% CL 26 - 37 mg/L

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

9.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

63 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: Behavior and appearance of test organism were observed in comparison with the control group. No abnormal behavior or appearance were observed in 6.2 mg/L and 9.3 mg/L groups. Abdominal swimming, lowering of mobility and immobilization were observed in 15.0 mg/L 24.6 mg/L, 39.2 mg/L, 63.0 mg/L and 97.7 mg/L groups.
- Mortality of control: none
- Concentration of the test substance in the test solution
For all concentration groups, concentrations of the test substance in the test solution were analyzed at the start of the exposure, at before and after of water change at 24 hours exposure and at the end of the exposure. Ratios of measured concentrations in each concentration group to the nominal concentration were 86 - 106 % in the test solution of at the start of the exposure and at after water change at 24 hours exposure and 90 - 112 % in the test solution of before water change at 24 hours exposure and at the end of the exposure. Ratios of measured concentrations in each concentration group to the initial concentration were 87 - 121 % in the test solution of before water change at 24 hours exposure and at the end of the exposure. Further average concentrations calculated by geometric average in each concentration groups were 6.2 mg/l for 6.5 mg/l group, 9.3 mg/l for 10 mg/l group, 15.0 mg/l for 16 mg/l group, 24.6 mg/l for 25 mg/l group, 39.2 mg/l for 40 mg/l group, 63.0 mg/l for 65 mg/l group and 97.7 mg/l for 100 mg/l group.
- Observation of condition of test solution: At the start of the exposure, it was confirmed by visual observation that the test solutions for all of the concentration groups were clear and colorless and the test substance was dissolved in the test solution. Further, at just before the water change after 24 hours exposure, there was no change in condition of the test solution.
- Validity of the test: On the basis of the fact that immobilization ratio was 0 % at the end of the exposure and no surfacing of Daphnia was observed during exposure for the control group, and dissolved oxygen concentration for each concentration group at the end of exposure were 3 mg/l or higher, the validity of the test was confirmed.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50: 0.43 mg/L

Reported statistics and error estimates:

EC50 were calculated by using Probit method. Further, 95 % confidence limits were also calculated and slope of regression line were obtained.
For calculation of results, statistical software "TOXDAT MULTI-METHOD PROGRAM (EPA/600/4-85/013)" was used.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this study it was found that HBPA has a EC50 31 mg/lL.

Description of key information

48 hour Daphnia Immobilisation:

EC₅₀ = 31 mg/L

EC₀ = 9.3 mg/L

EC₁₀₀= 63 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

31 mg/L

Additional information

A determination of the acute toxicity to aquatic invertebrates of HBPA was conducted (Ministry of the Environment Government of Japan, 2007, Study 18012 ). The study was performed in accordance with the Heisei 15/11/21-No.1121002 of the Pharmaceutical and Food Safety Bureau, Heisei 15/11/13-No.2 of the Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry and Heisei No.031121002 of the Environmental Policy Bureau, Ministry of the Environment, Japan, and in compliance with GLP.

20 less than 24 hours old Daphnia magna (4 replicates of 5 animals) were exposed to aqueous solutions of HBPA at nominal concentrations of 6.5 10, 16, 25, 40, 65 and 100 mg/L (calculated by geometric average 6.2, 9.3, 15.0, 24.6, 39.2, 63.0 and 97.7 mg/L) for 48 hours under semi-static test conditions.

Analysis of the fresh and old test preparations showed the measured concentrations to be within the range of 86 – 121 %; hence, the results are expressed in terms of calculated geometric average concentrations.

HBPA was found to be toxic to Daphnia magna under the conditions. The highest concentration at which no immobilisation occurred was 9.3 mg/L, whereas the lowest concentration at which 100% immobilisation occurred was 63.0 mg/L. It was concluded that the 48-hour EC50 in Daphnia magna was 31 mg/L.