3-(3,5-di-tert-butyl-4-hydroxyphenyl)-N-octadecylpropanamide

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 November 2011 to 16 January 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study undertaken at GLP accredited laboratory using internationally recognised method.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3-(3,5-di-tert-butyl-4-hydroxyphenyl)-N-C16-18-linear alkyl-propionamide

IUPAC Name:

3-(3,5-di-tert-butyl-4-hydroxyphenyl)-N-C16-18-linear alkyl-propionamide

Constituent 2

Reference substance name:

1235487-96-3

Cas Number:

1235487-96-3

IUPAC Name:

1235487-96-3

Constituent 3

Reference substance name:

T-1215A

IUPAC Name:

T-1215A

Details on test material:

- Name of test material (as cited in study report): T-1215A
- Substance type: Antioxidant for plastics.
- Physical state: White powder
- Analytical purity: 100%
- Purity test date: 18 August 2012
- Lot/batch No.: 10180U
- Expiration date of the lot/batch: 17 March 2012
- Stability under test conditions: No data
- Storage condition of test material: Room temperature
- Other: Sample received 30 March 2011

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0.01 mg/l
- Sampling method: Four, mid-vessel samples (10 mL) of media were taken from the control and test vessels at 0 and 72 hours (fresh media) and at 24 and 96 hours (expired media) for analysis.
- Sample storage conditions before analysis: On each occasion, two of the samples were analysed and the two others were stored in a freezer in case further analysis was required.

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The method of preparation used during the definitive test was based on the results of a range finding test. To minimise any loss of test substance by adsorption, glassware used in the study was either silanised or pre-conditioned to the test substance at 0.01 mg/L overnight before use. The pre-conditioning media were discarded immediately before the freshly prepared test solution was prepared.
Each day of the definitive test, the test substance (10 mg) was dissolved in THF (100 mL). An aliquot (100 μL/L) of this stock solution was added to diluent water (20 L) in a test vessel.
As an intermediate vehicle was used to facilitate the preparation of the test medium, an additional control group containing THF and dilution water (100 μL/L) was included in the study. The solvent control vessel was filled with dilution medium containing the same concentration of auxiliary substance as present in the test concentration.
- Eluate: Mobile phase A: Water:methanol (90:10 v:v) + 0.01M ammonium formate + 0.1% formic acid
Mobile phase B: Methanol:formic acid (100:0.1 v:v)
- Controls:Yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): tetrahydrafuran
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 100 µl / l
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No data

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: The fish were supplied by a commercial fish farm in the UK and were reared at the farm from eggs that hatched in September 2011.
- Age at study initiation (mean and range, SD): ca 4 months
- Length at study initiation (length definition, mean, range and SD): 5.52 cm
- Weight at study initiation (mean and range, SD): 1.50 g
- Method of breeding:
- Feeding during test: No

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions: Same as test
- Type and amount of food: The fish were fed daily with commercial fish food (TROUW (UK) Ltd; Nutra Fry 02)/Nishikoi Pellets/Tetramin Flake) an amount equivalent to between 1.0 and 1.5% of the total wet-weight of fish in the holding tank.
- Feeding frequency: Daily
- Health during acclimation: mortality was recorded as <0.5% in the 14 days before the definitive test.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Test conditions

Hardness:

Control; 173 mg/l CaCO3
Study; 143 mg/l CaCO3

Test temperature:

13.4 to 14.6ºC

pH:

8.02 to 8.38

Dissolved oxygen:

99 to 109% (air saturation value)

Salinity:

no data

Nominal and measured concentrations:

Nominal; 0.01 mg/l
Measured; 0.00870 mg/l (overall mean)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass
- Type: Aquarium
- Fill volume: 20 litres to a depth of 18.5 cm
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution: Daily batch renewal
- No. of organisms per vessel: 10
- No. of vessels per concentration:1!
- No. of vessels per control: 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: initial static loading of 0.75 g bodyweight/litre.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The water used to hold the fish and for the study was laboratory tap water, dechlorinated and softened by passage through a water purification system. It was passed through a high grade activated carbon filter to remove chlorine and any organic contaminants. A proportion of the supply then passed through a water softener before final reverse osmosis treatment to produce a highly purified water supply. The two grades of dechlorinated water were then remixed to give a supply with the desired water hardness. This water was then held in an intermediate tank where it was equilibrated to the test temperature and gently aerated before being supplied to the holding and test areas.
- Intervals of water quality measurement: 0, 24, 72 and 96 hours

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: A photoperiod of 16 hours light: 8 hours dark was maintained, with periods of subdued lighting at the beginning and end of each light phase.
- Light intensity: No data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :absence of respiratory movement and absence of response to physical stimulation of the caudal peduncle.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: None due to low solubility of the test substance
- Justification for using less concentrations than requested by guideline: low solubility of the test substance
- Range finding study: No

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.009 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.009 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: None
- Observations on body length and weight: No data

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

After 96 hours of exposure to T-1215A at its level of aqueous solubility, no mortality or significant sub-lethal effects were observed. Therefore, the 96 hour LC50 value was >0.00870 mg/L and the “no observed effect concentration” (NOEC) was ≥0.00870 mg/L.