cis-2-(bromomethyl)-2-(2,4-dichlorophenyl)-1,3-dioxolane-4-ylmethyl benzoate

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:

2017-02-03 to 2017-04-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Guidance document on aquatic toxicity testing of difficult items and mixtures, OECD series on testing and assessment number 23, December 14, 2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Analytical purity: impurities: R036844 3.30%, unknown <0.10%
- Source and lot/batch No.of test material:
* batch: I15DB1648
* test item: 207556/A
- Expiration date of the lot/batch: 2020-04-19 (retest date)
- Purity test date: 2015-05-27 (certificate of analysis release date)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: solubility in water: <0.05 g/L

Analytical monitoring:

yes

Details on sampling:

- Sampling method: samples for possible analysis were taken form both test groups 24 hours before the test, at the start of the test, after 48 and after 96 hours from the freshly prepared solutions. A volume of 2.0 mL was taken
- Sample storage conditions before analysis: the samples were stored in a freezer. Additionally, reserve samples of 2.0 mL were taken for possible analysis

Vehicle:

yes

Remarks:

Acetone

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
Exact volumes of the test item stock solutions and the solvent were dosed with syringes via a computer-controlled system consisting of two dispensers (Gilson). The dosed volumes (100 μL each 5 minutes) entered a mixing flask separately from the dilution water supply (adjusted ISO-medium). The dilution water was supplied applying flow-meters at a constant rate of 12 litres per hour. In the mixing flask the dosed volume and the dilution water were mixed under continuous stirring. The flow meters were calibrated before the start of the exposure. The whole system was checked at least once a day.       
* Range-finding test:
Due to an expected low water solubility of the test substance, a saturated solution (SS) was prepared. The preparation of test solutions started with a loading rate of 100 mg/L applying three days of magnetic stirring at room temperature to ensure maximum dissolution of the test item in the test medium. This resulted in a clear and colourless dispersion that contained undissolved material. The obtained mixture was filtered through a 0.45 µm membrane filter (Whatman; RC55). The filter was pre-conditioned with a small volume of test solution that was discarded. The resulting SS was used as the highest test concentration. Lower test concentrations were prepared by subsequent dilutions in test medium. All final test solutions were clear and colourless.
* Limit test:
A 0.24 mg/mL stock was prepared in acetone. No other treatment than vigorous shaking was needed to completely dissolve the test item in the solvent.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): All aqueous solutions were clear and colourless

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands
- Length at study initiation (length definition, mean, range and SD): range-finding test: 3.5 ± 0.3 cm, final test: 3.8 ± 0.3 cm
- Weight at study initiation (mean and range, SD): range-finding test: 0.56 ± 0.20 g, final test 0.66 ± 0.24 g
- Feeding during test: no

ACCLIMATION
- Acclimation period: at least 12 days after delivery
- Acclimation conditions (same as test or not): yes
- Type and amount of food: pelleted fish food
- Feeding frequency: daily
- Health during acclimation (any mortality observed): mortality less than 5% 7 days before test

Test type:

flow-through

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

not reported

Test temperature:

limit test: 20.9-21.2 °C

pH:

limit test: 8.1-8.3

Dissolved oxygen:

limit test: 8.8-9.6

Salinity:

not relevant

Nominal and measured concentrations:

Range-finding test:
100% SS at t=0h: measured concentration (mg/L) = 0.0787
100% SS at t=24h: measured concentration (mg/L) = 0.0042 (estimated value, calculated by extrapolation of the calibration curve)
100% SS at t=96h: measured concentration (mg/L) = n.d. (Not detected. The limit of detection of the method was determined to be 0.0031 mg/L taking a dilution factor of two into account)

Limit test:
Nominal concentrations (mg/L): 0, 0.024: a limit concentration being two times higher than solubility of the test item in water
Measured concentrations (mg/L) at t=-24h: n.d., 0.025
Measured concentrations (mg/L) at t=0h: n.d., 0.0243
Measured concentrations (mg/L) at t=48h: n.d., 0.0222
Measured concentrations (mg/L) at t=96h: n.d., 0.0227
n.d.: not detected

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 35L, all-glass
- Aeration: not during the test
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): not reported
- No. of vessels per control (replicates): not reported
- Biomass loading rate: 0.016 g fish/L/day

OTHER TEST CONDITIONS
- Adjustment of pH: 7.7 ± 0.3
- Photoperiod: 16h light daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Fish were considered to be dead when no reaction was observed after touching the caudal peduncle and visible breathing movements were absent. In addition, fish that were convulsing or showing other severe forms of distress not considered transient in nature and likely to become more severe before the exposure is terminated, will be sacrificed for humane reasons. These fish will be treated as having died in the test, based on OECD guideline on humane endpoint (ENV/JM/MONO/(2000)7).

TEST CONCENTRATIONS
- Spacing factor for test concentrations: none (limit test)
- Range finding study: yes
- Test concentrations: 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.024 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: none
- Other adverse effects control: no
- Abnormal responses: none

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality:
- EC50/LC50: 96-h LC50 = 0.32 mg/L. the sensitivity of the fish was within the range determined with the historical data

Reported statistics and error estimates:

No LC50 could be calculated because test item appeared to be not toxic (LC50 > maximum concentration tested).

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of JNJ-16479671-AAA (T001095) to Carp was determined in a 96-hour flow-through test according to the OECD guideline 203.
JNJ-16479671-AAA (T001095) did not induce mortality or other clinical effects at the limit concentration. Therefore, it can be concluded that the 96-h LC50 exceeded an analytically confirmed nominal concentration of 0.024 mg/L. The results of the test can be considered reliable without restriction.

Description of key information

The study of Tobor-Kaplon (2017), investigating the acute toxicity of T001095 in carp according to OECD guideline 203, was considered as the key study for endpoint coverage. An LC50 > 0.024 mg/L (based on analytically confirmed nominal test concentrations) was obtained after an exposure of 96 hours in a flow-through test in freshwater. Based on the results it can be considered that T001095 is not harmful to fish. The results of the test can be considered reliable without restriction.