Formamide, N-[(2-bromo-5-hydroxy-4-methoxyphenyl)methyl]-N-[2-(4-hydroxyphenyl)ethyl]-

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:

2017-02-17 to 2017-03-30

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: 98.9% (based on HPLC)
- Source and lot/batch No.of test material: I15EB1917
- Expiration date of the lot/batch: 2017-09-07 (retest date)
- Purity test date: 2017-06-07 (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: 0.01 g/L

Analytical monitoring:

yes

Details on sampling:

- Sampling method: singular samples for possible analysis were taken from the SS and the control at t=0 and t=96h. Volume was 2.0 mL.
- 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: Due to an expected low water solubility of the test item, 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 test medium. The obtained aqueous mixture was filtered through a 0.45 μm membrane filter (Whatman; RC55) to remove the undissolved fraction of the test item. 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 for the range-finding test were prepared by subsequent dilutions of the SS in test medium.
- Controls: yes
- Evidence of undissolved material (e.g. precipitate, surface film, etc): All final test 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: 3.4 ± 0.2 cm, limit test: 3.8 ± 0.1 cm
- Weight at study initiation (mean and range, SD): range-finding test: 0.59 ± 0.16 g, limit test 0.71 ± 0.12 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% seven days before test

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

not reported

Test temperature:

21 °C

pH:

7.3-8.0

Dissolved oxygen:

6.7-8.8

Salinity:

not relevant

Conductivity:

not indicated

Nominal and measured concentrations:

Range-finder:
Nominal concentrations: 1, 10 and 100% of SS (1 and 10% of SS not analysed)
Measured concentrations at t=0h: 7.34 mg/L
Measured concentrations at t=24h: 7.27 mg/L
Measured concentrations at t=96h: 7.17 mg/L

Final test:
Nominal concentrations: SS prepared at 100%
Measured concentrations at t=0h: n.d., 8.08 mg/L
Measured concentrations at t=96h: n.d., 7.93 mg/L

n.d. = not detected

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 10 L, all-glass filled with 9.8 L 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: 0.51 g fish/L

OTHER TEST CONDITIONS
- 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.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: x10 (limit test)
- Range finding study: yes
- Test concentrations: 1, 10 and 100% of a SS at 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.:

> 8.1 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: none
- Other adverse effects control: no
- Mortality observed at the limit test concentration: none
- Other effects observed at the limit test concentration: none

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50/LC50: 96-h LC50 = 0.32 mg/L.

Reported statistics and error estimates:

No LC50 could be calculated because the effects were below 50% (LC50 > maximum soluble concentration tested).

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of T002078 to carp was determined in a 96-hour static limit test according to OECD guideline 203. Under the conditions of the present study, the 96-h LC50 was >8.1 mg/L based on the initial exposure concentration. The test item had no mortal effect on Carp after an exposure of 96 hours at the maximum solubility of test item in test medium. The results of the test can be considered reliable without restriction.

Description of key information

The study of Bouwman (2017), investigating the acute toxicity of T002078 in carp according to OECD guideline 203, was considered as the key study for endpoint coverage. A LC50 > 8.1 mg/L (based on initial exposure concentration and being considered the maximum solubility of test item in test medium) was obtained after an exposure of 96 hours in a limit static test in freshwater. Based on the results it can be considered that T002078 is not harmful to fish.