cis-2-(2,4-dichlorophenyl)-2-(1H-imidazol-1-ylmethyl)-1,3-dioxolane-4-ylmethyl methanesulphonate monohydrochloride

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:

2015-09-21 to 2015-11-13

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 substances 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
- Source and lot/batch No.of test material: I14KB4717
- Expiration date of the lot/batch: 2016-11-25 (retest date)
- Purity test date: 2014-12-13 (certificate of analysis release date)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Stability under test conditions: no data

Analytical monitoring:

yes

Details on sampling:

- Sampling method: samples for possible analysis were taken form both test groups at the start of the test and after 72 hours from the freshly prepared solutions. At the first renewal (t=24h) and at the end of the test from the 24h old solutions. 2.0 mL of volume 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:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
Range-finding test:
Based on the data provided by the Sponsor, the solubility of the test substance in water was 39.3 g/L. Therefore, the preparation of test solutions started with a loading rate of 100 mg/L (duplicate samples) applying a short period of magnetic stirring (47-51 minutes at room temperature) where after the solutions were hazy and contained flocks. These solutions were discarded. New solution was prepared in Milli-RO water (100 mg/L); after a 3-minute period of magnetic stirring the solution was clear and colourless confirming sufficient solubility of the test item in water. Subsequently salts were added to reach the concentrations described in paragraph 6.3 of the report (adjusted ISO medium) and used as the highest concentration. The lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium. The final test solutions were all clear and colourless at the start of the test.
Due to appearance of undissolved material during the study and the decrease of the measured concentration in the highest treatment, it was concluded that the test item reacts with salts present in the medium. Therefore, it was decided, in consultation with the sponsor, to change the preparation
procedure in the final test.

Final test:
Based on the observations made in the range-finding test and in agreement with the sponsor, the solutions for the final test were prepared differently. A Saturated Solution (SS) was prepared at a loading rate of 100 mg/L (in duplicate) applying a one-hour period of magnetic stirring. The obtained mixture was filtered through a preconditioned membrane filter (RC55, Whatman). Filtrates originating from both replicates were combined and used as the highest test concentration. The lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium. The final test solutions were all clear and colourless at the start of the test.
- 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: 2.0 ± 0.1 cm, final test: 2.1 ± 0.1 cm
- Weight at study initiation (mean and range, SD): range-finding test: 0.16 ± 0.04 g, final test 0.13 ± 0.03 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:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

not reported

Test temperature:

21.3-21.7°C

pH:

7.0-7.7

Dissolved oxygen:

7.0-9.3

Salinity:

not relevant

Nominal and measured concentrations:

Range-finder:
Nominal concentrations (mg/L): 0.1, 1, 10, 100 mg/L (samples from 10 and 100 mg/L were analysed)
Measured concentrations (mg/L) at t=0h: 11, 100
Measured concentrations (mg/L) at t=24h: 10, 81
Measured concentrations (mg/L) at t=96h: 10, 52

Final test:
Nominal concentrations (mg/L): 4.6, 10, 22, 46 and 100% of a SS prepared at 100 mg/L
Measured concentrations (mg/L) at t=0h: 0.0043, 3.96, 8.57, 18.8, 38.0, 85.1
Measured concentrations (mg/L) at t=24h: 37.9
Measured concentrations (mg/L) at t=96h: 0.0048, 3.94, 8.60, 18.8

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 6.5L, all-glass filled with 3L test solution
- 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: 0.30 g fish/L

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
- Range finding study: yes
- Test concentrations: 0.10, 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.:

31 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.L.: 25-39 mg/L

Details on results:

- Mortality of control: none
- Other adverse effects control: no
- Abnormal responses: Hypoactive, Discoloured and swimming at the surface, Loss of equilibrium

Results with reference substance (positive control):

- Results with reference substance valid? yes
- 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:

The LC50 was determined using the maximum likelihood estimation method with the probits of the percentages of dead fish as function of the logarithms of the corresponding concentrations. Calculations were performed with ToxRat Professional v. 3.2.0. (ToxRat Solutions®, GmbH, Germany).

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of T001202 to Carp was determined in a 96-hour static test according to the OECD guideline 203.
The 96-h LC50 was 31 mg/L based on analytically confirmed nominal concentrations (95% confidence interval between 25 and 39 mg/L) and was reached already after 24 hours of exposure. The results of the test can be considered reliable without restriction.

Description of key information

The study of Tobor-Kaplon (2016), investigating the acute toxicity of T001202 in carp according to OECD guideline 203, was considered as the key study for endpoint coverage. The 96-h LC50 was 31 mg/L based on analytically confirmed nominal concentrations (95% confidence interval between 25 and 39 mg/L) and was reached already after 24 hours of exposure. The results of the test can be considered reliable without restriction.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

31 mg/L