Diallyl hexahydrophthalate

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:

29th January 2018 - 12th September 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

Adopted July 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Source: Osaka Soda Co. Ltd. Batch/lot number: 40201
- Expiration date of the lot/batch: 26 January 2019
- Purity test date: 99.2%

RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: n/a
- Specific activity: n/a
- Locations of the label: n/a
- Expiration date of radiochemical substance: n/a

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, protected from light
- Stability under test conditions: Not specified
- Solubility and stability of the test substance in the solvent/vehicle: Not specified
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: n/a

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: A stock solution with a concentration of 100 mg/L (nominal) was prepared by dissolving an appropriate amount of Test Item in aquarium water.
- Preliminary purification step (if any): n/a
- Final dilution of a dissolved solid, stock liquid or gel: 100 mg/L stock solution from which test solutions were prepared by appropriate dilution of this stock solution just before introduction of the fish
- Final preparation of a solid: n/a

FORM AS APPLIED IN THE TEST (if different from that of starting material) : n/a

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable) : n/a

OTHER SPECIFICS: n/a

Analytical monitoring:

no

Remarks:

Test item concentration in the test solutions was determined at the beginning and at the end of the renewal periods. The samples were analysed by a HPLC-UV method.

Details on sampling:

n/a

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: n/a
- Eluate: n/a
- Differential loading: n/a
- Controls: n/a
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): n/a
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): n/a
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): n/a

Test organisms (species):

other: Brachydanio rerio (Zebrafish)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Strain: Brachydanio rerio
- Source: Szent István University, Department of Aquaculture, Hungary
- Age at study initiation (mean and range, SD): Not specified
- Length at study initiation (length definition, mean, range and SD): 2.5 - 2.9 cm
- Weight at study initiation (mean and range, SD): Not specified
- Method of breeding: Not specified
- Maintenance of the brood fish: Housed under disease- and parasite-controlled conditions,

ACCLIMATION
- Acclimation period: >12 days
- Acclimation conditions (same as test or not): Same
- Type and amount of food during acclimation: Not specified
- Feeding frequency during acclimation: Not specified
- Health during acclimation (any mortality observed): Mortality was less than 5% of the population for seven days before the test.

QUARANTINE (wild caught)
- Duration: n/a
- Health/mortality: n/a

FEEDING DURING TEST
- Food type: Not specified
- Amount: Not specified
- Frequency: Not specified

Test type:

semi-static

Water media type:

not specified

Remarks:

Described as being "aquarium water"

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

Exposure period of 96 hours. The frequency of the renewal periods were 48 hours. All fish were observed to be dead after 48 hours (first renewal) at the two highest concentrations of 5.0 and 10.0 mg/L (nominal).

Post exposure observation period:

96 hour exposure duration with one renewal at 48 hours and a second at 96 hours. No further observation period after the end of the 96 hour exposure duration.

Hardness:

191 mg/L (as CaCO3)

Test temperature:

20.2 - 21.1 °C

pH:

7.34 – 8.00

Dissolved oxygen:

71 – 100 % of the air saturation value at the temperature used

Salinity:

Not specified

Conductivity:

Not specified

Nominal and measured concentrations:

Nominal: 0.63, 1.25, 2.50, 5.0 and 10.0 mg/L
Measured: 0.59, 1.09, 2.07, 4.81 and 9.76 mg/L As the measured concentrations deviated more than 20% from the nominal in few cases, the biological results are based on the measured concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquarium
- Type (delete if not applicable): n/a
- Material, size, headspace, fill volume: 5L volume of test solution
- Aeration: Not specified
- Type of flow-through (e.g. peristaltic or proportional diluter): Not specified
- Renewal rate of test solution (frequency/flow rate): 48 hours
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): n/a
- Biomass loading rate: 1.0 g fish/L test solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Not specified
- Total organic carbon: Not specified
- Particulate matter: Not specified
- Metals: Not specified
- Pesticides: Not specified
- Chlorine: Not specified
- Alkalinity: Not specified
- Ca/mg ratio: Not specified
- Culture medium different from test medium: No
- Intervals of water quality measurement: Water temperature, oxygen concentration and pH were measured at least daily in each test aquarium.

OTHER TEST CONDITIONS
- Adjustment of pH: Not specified
- Photoperiod: 16 hours light and 8 hours darkness
- Light intensity: Not specified

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality, body length and body weight. The observations of fish were carried out in the following intervals: at 3, 6, 24, 48, 72 and 96 hours.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0
- Justification for using less concentrations than requested by guideline: Not specified
- Range finding study : Yes
- Test concentrations: Control, 0.1, 1.0, 10.0 and 100.0 mg/L
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.98 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

ca. 1.23 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

ca. 1.71 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

ca. 3.75 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

ca. 2.07 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 0.59 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

ca. 1.09 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities:
- Observations on body length and weight: The body weights of the fish were recorded before the introduction of fish. The body weight of 7 fish per aquarium was registered. There was no considerable difference observed concerning body weights between the groups.
- Other biological observations: Behaviour of fish was found as normal at the lowest concentration level and in the Control as well. Dark colour, localization close to the surface of the water were observed in the 1.09 mg/L (measured) concentration. Dark colour, localization close to the surface of the water and to the bottom of the aquarium, decreased activity, diagonal swimming, and slow motility of the operculum were observed in the 2.07 mg/L (measured) concentration. Localization close to the surface of the water and decreased activity were observed in the 4.81 mg/L (measured) concentration. Localization at the bottom of the aquarium, localization close to the surface of the water, decreased activity, unbalanced swimming, diagonal swimming, lateral position and slow motility of the operculum and swimming in the same place were observed at the highest concentration level during the experiment.
- Mortality of control: No
- Other adverse effects control: No
- Abnormal responses: No specified
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Not specified
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

n/a

Reported statistics and error estimates:

n/a

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this acute fish toxicity study on Zebrafish (Brachydanio rerio) the observed endpoints for the effect of the Test Item were the following:
24h LC50 value - 3.75 mg/L
48h LC50 value - 1.71 mg/L
72h LC50 value - 1.23 mg/L
96h LC50 value - 0.98 mg/L
96h LC100 value - 2.07 mg/L
96h NOEC - 0.59 mg/L
96h LOEC - 1.09 mg/L

Executive summary:

The acute toxicity of the Test Item MDAC was assessed with acute fish toxicity test on Zebrafish (Brachydanio rerio),over an exposure period of 96 hours in a semi-static system. The frequency of the renewal periods were 48 hours. Based on the preliminary range-finding test, 5 test concentrations in a geometric series with a separation factor of 2.0 and 1 untreated Control were tested in the main experiment, these being 0.59; 1.09; 2.07, 4.81 and 9.76 mg/L. As the measured concentrations deviated more than 20% from the nominal in few cases, the biological results are based on the measured concentrations. One aquarium was used in the test groups and one in the Control group. Each aquarium comprised 7 fish and 5 litre test solution. All achievable validity criteria were met during this study.

Under the conditions of this acute fish toxicity study on Zebrafish (Brachydanio rerio) the observed endpoints for the effect of the Test Item were the following:

24h LC50 value - 3.75 mg/L

48h LC50 value - 1.71 mg/L

72h LC50 value - 1.23 mg/L

96h LC50 value - 0.98 mg/L

96h LC100 value - 2.07 mg/L

96h NOEC - 0.59 mg/L

96h LOEC - 1.09 mg/L

Description of key information

This substance was found to have a 96h LC50 0.98 mg/l

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.98 mg/L