N,N'-(ethoxymethylsilylene)bis[N-methylbenzamide]

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:

September 4, 2017 - September 8, 2017

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:

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:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature.

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 (control) and 100 mg/L.
- Sampling method: The test concentration was analytically determined at the start and at the end of the renewal periods (0, 48, 96h) by HPLC-UV method.
- Sample storage conditions before analysis: Formulation samples were stored at test conditions for different time intervals. At the end of the storage period formulations were analysed.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: a stock solution with a nominal concentration of 100 mg/L was prepared with direct addition of the test item, mixed into the test medium (ISO medium) using ultrasonic bath (aprox. 30 minutes). As a Limit test was carried out, further dilution of stock solution was not performed.
- Controls: The dilution water (ISO medium) was used without addition of the test item
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): no.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish.
- Strain: Brachydanio rerio.
- Source: István Károly u. 1, Hungary.
- Age at study initiation (mean and range, SD): not specified.
- Length at study initiation (length definition, mean, range and SD): 2.6 - 2.9 cm.
- Weight at study initiation (mean and range, SD): 1.77 - 1.80 g.

ACCLIMATION
- Acclimation period: more than 12 days.
- Acclimation conditions (same as test or not): same as test.
- Health during acclimation (any mortality observed): Fish were bred in a well-known fish farm, under desease- and parasite-controlled conditions. Fish were observed during the acclimatisation and test periods, the fish were healthy. There was no mortality of the population for seven days before the test.

FEEDING DURING TEST
- The fish were not fed during the test.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

187 mg/L (as CaCO3)

Test temperature:

20.4 - 24.5 ºC

pH:

7.75 - 8.11

Dissolved oxygen:

86 - 97% of the air saturation value at the temperature used.

Nominal and measured concentrations:

- Nominal concentrations: 0 (control) and 100 mg/L.
- Measured concentrations: 0 (control), 103 ± 1.92 mg/mL (at the beginning of thet est) and 110 ± 1.38 mg/L (at the end of the test). As the measured concentration deviated not more than 20 per cent from the nominal, the biological results are based on the nominal concentration.

Details on test conditions:

TEST SYSTEM
- Test vessel: one aquarium was used in the test group and one in the control group.
- Material, size, headspace, fill volume: fill volume 5 L test solution.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: The loading of the test aquaria was less than 1.0 g fish/L test solution at the start of the experiment.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: aquarium water (circulated and filtered).
- Culture medium different from test medium: no.

OTHER TEST CONDITIONS
- Adjustment of pH: no.
- Photoperiod: 16 hours light and 8 hours darkness.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The observations of fish were carried out in the following intervals. at 3,6, 24, 48, 72 and 96 hours. Mortality or any sub-lethal effects were not observed durint the experiment. The test conditions (pH, temperature, oxygen saturation) were measured daily during the test. The body weights of the fish were recorded before the introduction of fish. The body weight of 7 fish per aquarium was registered. The loading of the aquaria was calculated on the basis of these body weights (1.0 g fish/liter testing liquid).

TEST CONCENTRATIONS
- Range finding study : Four fish were exposed to each concentration of the test item plus a control, for 96 hours under semi-static conditions (with 24-hour renewal in absence of stability data).
- Test concentrations: 0 (control), 0.1, 1, 10 and 100 mg/L test item.
- Results used to determine the conditions for the definitive study: Because no toxic response was observed during the preliminary range-finding test, only one test concentration (100 mg/L) and one control group was tested in the definitive study in a static system.

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: No abnormal behaviour, appearance of any other effect on test animals were detected.
- Observations on body length and weight: There was no considerable difference observed concerning body weights between the groups. Body lengths were within the acceptable ranges.
- Mortality of control: 0/7.
- Other adverse effects control: no.
- Abnormal responses: no.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no.
- Effect concentrations exceeding solubility of substance in test medium: no.

Reported statistics and error estimates:

No statistical analysis was performed because of the lack of toxic effects.

Sublethal observations / clinical signs:

Table 2:Cumulative mortality data in the Limit Test

Test Group	Cumulative Mortality (number of dead fish) [initial population = 7 fish]
	3h	6h	24h	48h	72h	96h
Control	0	0	0	0	0	0
100 mg/L (nominal)	0	0	0	0	0	0

 

Table 3:Measured and calculated data of bodyweight

Test Group	Measured weight of 7 fish (g)	Calculated mean weight of 1 fish (g)	Loading of testing aquarium (g fish/L testing liquid)
Control	1.77	0.25	0.35
100 mg/L (nominal)	1.80	0.26	0.36

Table 4:Temperature

Test Group	Temperature (°C)
	Measuring
	0h	24	48	72	96
Control	21.3	20.6	20.4	20.6	20.5
100 mg/L (nominal)	24.5	20.6	20.5	20.7	20.6

 

 

Table 5:Dissolved oxygen concentration

Test Group	Dissolved oxygen concentration (%)
	Measuring
	0h	24	48	72	96
Control	97	92	94	94	93
100 mg/L (nominal)	90	86	93	92	89

 

 

Table 6:pH

Test Group	pH
	Measuring
	0h	24	48	72	96
Control	7.98	7.85	7.75	7.83	7.68
100 mg/L (nominal)	8.11	8.00	7.99	8.05	7.86

Table 7:Body length of test fish at the end of the test

Test Group	Body length [cm]
	1	2	3	4	5	6	7
Control	2.7	2.6	2.9	2.8	2.7	2.8	2.7
100 mg/L (nominal)	2.9	2.8	2.7	2.7	2.8	2.7	2.9

 Remark: At the end of the test,body length of test fish were measured in order to check their compliance with the size range recommended for the species by the test guideline [Commission Regulation (EC) No 440/2008;Annex Part C, C.1]

Validity criteria fulfilled:

yes

Remarks:

All validity criteria were fulfilled, see "Overall remarks".

Conclusions:

The 96h-EC50 of the test item in Zebrafish (Brachydanio rerio) was determined to be higher than 100 mg/L and the NOEC was 100 mg/L, both based on mortality.

Executive summary:

An acute toxicity test on zebrafish (Brachydanio rerio) was performed in accordance with OECD Guideline 203 and EU method C.1, following GLP. Based on the results obtained in a preliminary range-finding test where no toxic response was observed, a Limit Test was carried out using only one test concentration (100 mg/L) and one control group in a static system. The test concentrations were analytically determined (validated method) at the beginning and at the end of each renewal period. As the measured concentration did not deviate more than 20 percent from the nominal, the biological results are based on the nominal concentration. Seven fish per group were exposed to the test item or the control for 96 hours. No mortality or other abnormalities were observed and all validity criteria were met. Consequently, the 96h-EC50 of the test item in zebrafish (Brachydanio rerio) was determined to be higher than 100 mg/L and the NOEC was 100 mg/L, both based on mortality.

Description of key information

Key study: OECD 203 and EU method C.1. GLP study. The 96h-EC50 of the test item in zebrafish (Brachydanio rerio) was determined to be higher than 100 mg/L and the NOEC was 100 mg/L, both based on mortality.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Key study: An acute toxicity test on zebrafish (Brachydanio rerio) was performed in accordance with OECD Guideline 203 and EU method C.1, following GLP. Based on the results obtained in a preliminary range-finding test where no toxic response was observed, a Limit Test was carried out using only one test concentration (100 mg/L) and one control group in a static system. The test concentrations were analytically determined (validated method) at the beginning and at the end of each renewal period. As the measured concentration did not deviate more than 20 percent from the nominal, the biological results are based on the nominal concentration. Seven fish per group were exposed to the test item or the control for 96 hours. All validity criteria were met. The 96h-EC50 of the test item in zebrafish (Brachydanio rerio) was determined to be higher than 100 mg/L and the NOEC was 100 mg/L, both based on mortality.