1-Propanol, 3-[[3-[[[3-(acetyloxy)-2,2-dimethylpropylidene]amino]methyl]-3,5,5-trimethylcyclohexyl]imino]-2,2-dimethyl-, 1-acetate

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:

from 2017-01-19 to 2017-03-09

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:

Batch No.: I1601867-MG-06-120.1
Purity: 95.7%

Analytical monitoring:

yes

Details on sampling:

- Sampling method: For determination of the test item concentration five replicate samples were taken from the test solution and one from the control at the start and at the end of the exposure phase of the test.

Vehicle:

no

Details on test solutions:

The test solution used in the test was prepared by mechanical dispersion without using any solubilising agent. 0.5 g of test item was dissolved in 5000 mL of the dilution water (ISO medium) in order to obtain the concentration of 100 mg/L, thereafter stirred for 20 hours at 300 rpm. The resulted solution was the test concentration.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: Akvárium Magazin Kft., 1222 Budapest Dévény u. 36., Hungary
- Age at study initiation: juveniles
- Length at study initiation: The body length of each fish were measured at the end of the test in order to check their compliance with the size range recommended for the species by the test guideline (OECD No. 203). The measured values were within the range of 2.25 -2.90 cm.
- Weight at study initiation: The weight of the group of fish was registered and the average body weight was calculated for 1 fish. It was in the range of 0.29 - 0.32 g. The loading of the test aquariums was also calculated and was 0.58 g/L in the control and 0.63 g/L in the test item treated group.
- Method of breeding: Fish are bred in a well-known fish farm, under disease- and parasite-controlled conditions.

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions: same as the test conditions during the exposure period
- Type and amount of food during acclimation: Fish were fed with appropriate, commercial diet for fish.
- Feeding frequency during acclimation: at least three times per week until 3 days before treatment
- Health during acclimation: No mortality occurred in seven days before the start of the experiment, therefore the batch was considered to be acceptable for testing.

FEEDING DURING TEST
- No feeding during the exposure period.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No post exposure observations were performed.

Hardness:

249 mg/L CaCO3

Test temperature:

22.4 - 23.2 °C

pH:

6.73 - 7.21

Dissolved oxygen:

60.9 - 103.8 % of the air saturation value

Salinity:

NA

Conductivity:

NA

Nominal and measured concentrations:

Nominal concentration: 100 mg/L
Measured concentration: 94.0 - 97.0 mg/L (at the start of the test), 89.9 - 96.0 mg/L (at the end of the test)

Details on test conditions:

TEST SYSTEM
- Test vessel: aquariums
- Type: open
- Material, size, headspace, fill volume: Glass, approx. 6 litres capacity
- Aeration: slight aeration in the test solution after oxygen concentration measurement on the 48-hour, till the end of the exposure period
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
The ISO medium was prepared by adding 25 mL from each of four stock solutions to one litre deionised water proportionally.
Stock solution 1: CaCl2 x 2H2O 11.76 g/L
Stock solution 2: MgSO4 x 7H2O 4.93 g/L
Stock solution 3: NaHCO3 2.59 g/L
Stock solution 4: KCl 0.23 g/L

OTHER TEST CONDITIONS
- Adjustment of pH: No adjustment of the pH was carried out.
- Photoperiod: 16 h light: 8 h dark cycle

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: Only one nominal concentration (100 mg/L) was used, based on the results prom a preliminary test, where no mortality was observed in both the control and the test solution (100 mg/L). Thus, as a main test, a limit test with one test group was performed.
- Test concentrations: 100 mg/L

Reference substance (positive control):

no

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:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

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:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: There were no mortalities or any sub-lethal effects observed in 10 fishes in the control group or in 10 fishes exposed to the 100 mg/L nominal concentration.
- Mortality of control: No mortality was observed in the control group during the test.
- Abnormal responses: None were observed.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None were observed.

Reported statistics and error estimates:

Since a limit test was performed (and mortality was not occurred), no statistical analysis was necessary. The LC0, LC50, NOEC, LOEC and the LC100 values were determined directly from the raw data.

Sublethal observations / clinical signs:

Table 1: Results of the PreliminaryRange-Finding Test

Nominal concentrations [mg/L]	Untreated control	0.1	1	10	50	100
Number of treated animals	3	3	3	3	3	3
Number of dead animals (at 96h)	0	0	0	0	0	0

 

 

Validity criteria fulfilled:

yes

Conclusions:

The test item SIKA Hardener AI had no toxic effect up to at least the limit concentration of 100 mg/L on Zebrafish. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L.

Executive summary:

The purpose of a 96-hour study was to evaluate the acute toxicity of the test item SIKA Hardener AI on Zebrafish (Brachydanio rerio). A limit test was performed in which the test animals were exposed to aqueous test media containing the test item for 96 hours at only one concentration (100 mg/L) plus a control in order to demonstrate that the test item is not toxic to Zebrafish up to at least this test concentration (i.e. limit concentration). Based on the results obtained during analytical method validation, the test item hydrolyzes very quickly, therefore, concentration of the hydrolysis product was determined. The hydrolysis product, 5-Amino-1,3,3-trimethylcyclohexanemethylamine is stable for 96 hours in ISO medium, therefore a static test was performed.

For this purpose, young fish were exposed in a static test to aqueous test media containing the test item for 96 hours. Based on the results of the preliminary test, the main test was a limit test performed using a treatment group at saturation (corresponding to 100 mg/L nominal concentration) and one concurrent control group. The test was run according to international test guidelines - OECD 203 "Fish, Acute Toxicity Test", EU Method C. 1 "Acute Toxicity for Fish" and EPA OPPTS 950.1075, using the recommended fish species Zebrafish (Brachydanio rerio). Mortality and any sub-lethal effects were not observed during the 96-h exposure period either in the treated or in the control group. Accordingly, the 96-h NOEC was determined as 100 mg/L and the LOEC and the LC₅₀values were determined to be higher than 100 mg/L.

Description of key information

The test item SIKA Hardener AI had no toxic effect up to at least the limit concentration of 100 mg/L on Zebrafish. Accordingly, the 96-h LC₅₀ value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L.

Additional information

The purpose of a 96-hour study was to evaluate the acute toxicity of the test item SIKA Hardener AI on Zebrafish (Brachydanio rerio). A limit test was performed in which the test animals were exposed to aqueous test media containing the test item for 96 hours at only one concentration (100 mg/L) plus a control in order to demonstrate that the test item is not toxic to Zebrafish up to at least this test concentration (i.e. limit concentration). Based on the results obtained during analytical method validation, the test item hydrolyzes very quickly, therefore, concentration of the hydrolysis product was determined. The hydrolysis product, 5-Amino-1,3,3-trimethylcyclohexanemethylamine is stable for 96 hours in ISO medium, therefore a static test was performed.

For this purpose, young fish were exposed in a static test to aqueous test media containing the test item for 96 hours. Based on the results of the preliminary test, the main test was a limit test performed using a treatment group at saturation (corresponding to 100 mg/L nominal concentration) and one concurrent control group. The test was run according to international test guidelines - OECD 203 "Fish, Acute Toxicity Test", EU Method C. 1 "Acute Toxicity for Fish" and EPA OPPTS 950.1075, using the recommended fish species Zebrafish (Brachydanio rerio). Mortality and any sub-lethal effects were not observed during the 96-h exposure period either in the treated or in the control group. Accordingly, the 96-h NOEC was determined as 100 mg/L and the LOEC and the LC₅₀ values were determined to be higher than 100 mg/L.