4,4'-isopropylidenedicyclohexanol

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 March 2007 to 23 March 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: Joint Notification of Heisei 15/11/21-No.1121002 of the Pharmaceutical and Food Safety Bureau

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Ministry of Health, Labour and Welfare, Heisei 15/11/13-No.2 of the Manufacturing Industries Bureau

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Ministry of Economy, Trade and Industry and No.031121002 of the Environmental Policy Bureau, Ministry of the Environment, Japan

Deviations:

no

GLP compliance:

yes

Remarks:

Note that this test was conducted in conformity to “The Standards Concerning Test Facilities Conducting the Tests Relating to New Chemical Substances”, Heisei 15/11/21-No.1121003, Heisei 15/11/17-No.3 and No.031121004.

Analytical monitoring:

yes

Details on sampling:

For all concentration groups, total four times, i.e. at the start of the exposure, at water change after 24 hours exposure, at water change after 72 hours exposure and at the end of the exposure, concentrations of the test substance were analyzed. Note that 100 mL of the test solution were taken from the middle layer of the test vessel and were applied as 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 24 °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 24 °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):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Medaka
- Source: Japanese fish farm (Kounosu City, Saitama, Japan)
- Age at study initiation: 2 months
- Length at study initiation: 1.9 - 2.0 cm
- Weight at study initiation: 0.07 - 0.08 g
- Feeding during test: no

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions: Same as test
- Type and amount of food: Commercially-supplied blend feed "Sodateeru" from Chubushiryo CO., LTD, about 3 % of body weight per day
- Feeding frequency: Twice a day, every day. For 24 hours before the exposure start, no feed was given to the fish.
- Health during acclimation: no mortalities

Test type:

semi-static

Water media type:

other: Dechlorinated water

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Observation of life/death and disease presentation, etc. of test organism was conducted at time elapse of 3, 6, 24, 48, 72 and 96 hours after the start of exposure

Hardness:

54 - 60 mg CaCO3/ L

Test temperature:

23.1 - 23.9°C

pH:

7.6 - 8.0

Dissolved oxygen:

7.5 - 8.6 mg/L (During exposure period, dissolved oxygen concentration was kept at 60 % or more of the saturated dissolved oxygen concentration. Aeration was not applied)

Nominal and measured concentrations:

Nominal: 10, 13, 18, 24 and 32 mg/L
Measured: 8.9, 12.4, 15.5, 21.1 and 29.5 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 5L glass
- Type: Loosely covered to reduce the loss of water due to evaporation and to avoid the entry of dust into the solutions.
- Fill volume: 4L
- Aeration: no
- Type of flow-through: Semi-static
- Renewal rate of test solution: Every 24 hour
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

OTHER TEST CONDITIONS
- Photoperiod: Room light, 16 hours light, 8 hours dark
- Light intensity: no data
- pH: 7.6 - 8. 0 (pH conditioning was not applied

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.3
- Range finding study: yes
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

copper sulfate [II] pentahydrate,

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 19 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL (17 - 21 mg/L)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

12.4 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

29.5 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

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 8.9 mg/l and 12.4 mg/l groups. Abdominal swimming, abdominal distention, immobility, overturn and surfacing were observed in 15.5 mg/l and 21.1 mg/l groups and abdominal distention, overturn and surfacing were observed in 29.5 mg/l group.
- Observations on body length and weight: None after test.
- Mortality of control: No such behaviour was observed in the control group.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no
- Concentration of the test substance in the test solution: Ratios of measured concentrations in each concentration group to the nominal concentration were 85 - 108 % in the test solution of at the start of the exposure and at water change after 72 hours exposure and 81 - 92 % in the test solution of at water change after 24 hours exposure and at the end of the exposure. Ratios of measured concentrations in each concentration group to the initial concentration were 75 - 99 % in the test solution of at water change after 24 hours exposure and at the end of the exposure.
Average concentrations calculated by geometric average in each concentration groups were 8.9 mg/l for 10 mg/l group, 12.4 mg/l for 13 mg/l group, 15.5 mg/l for 18 mg/l group, 21.1 mg/l for 24 mg/l group and 29.5 mg/l for 32 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, immediately 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 lethal ratio was 0 % at the end of the exposure and dissolved oxygen concentration for each concentration group were 60 % or more of the saturated dissolved oxygen concentration at test water temperature, the validity of the test was confirmed.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- LC50: 1.1 mg/L

Reported statistics and error estimates:

- LC50 was calculated by using Binominal method and Probit method. To a maximum extent, 95 % confidence limits were also calculated and gradients of regression line were obtained. Further, a graph of concentration-mortality was also made.
- For calculation of results, statistical software "TOXDAT MULTI-METHOD PROGRAM (EPA/600/4-85/013)"

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this study it was found that HBPA has a LC50 19 mg/L.

Description of key information

96 hour Acute Fish:

LC₅₀= 19 mg/L

LC₀ = 12.4 mg/L

LC₁₀₀ = 29.5 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

19 mg/L

Additional information

A determination of the acute toxicity to fish of HBPA was conducted (Ministry of the Environment Government of Japan, 2007, Study 18014). The study was performed in accordance with Joint Notification of Heisei 15/11/21-No.1121002 of the Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare, Heisei 15/11/13-No.2 of the Manufacturing Industries Bureau and Ministry of Economy, Trade and Industry and No.031121002 of the Environmental Policy Bureau, Ministry of the Environment, Japan, and in compliance with GLP.

Groups of 10 Medaka (Oryzias latipes) were exposed to aqueous solutions of HBPA at nominal concentrations of 10, 13, 18, 24 and 32mg/L (calculated geometric average 8.9, 12.4, 15.5, 21.1 and 29.5 mg/L) for 96 hours at temperature of 23 - 24 °C, under semi-static test conditions.

Analysis of the fresh and old test preparations showed the measured concentrations to be within the range of 75 – 108 %; hence, the results are expressed in terms of calculated geometric average concentrations.

HBPA was found to be toxic to Oryzias latipes under the conditions. The highest concentration at which no mortality occurred was 12.4 mg/L, whereas the lowest concentration at which 100% mortality occurred was 29.5 mg/L. It was concluded that the 96-hour EC50 in Oryzias latipes were 19.0 mg/L.