A mixture of: esters of C14-C15 branched alcohols with 3,5-di-t-butyl-4-hydroxyphenyl propionic acid; C15 branched and linear alkyl 3,5-bis(1,1-dimethylethyl)-4-hydroxybenzenepropanoate; C13 branched and linear alkyl 3,5-bis(1,1-dimethylethyl)-4-hydroxybenzenepropanoate

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to fish, other

Type of information:

experimental study

Adequacy of study:

key study

Study period:

4 June 1997 - 25 June 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

not specified

Details on test solutions:

Identity and concentration of auxiliary solvent for dispersal: Acetone: 33 µl/l

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Brachydanio rerio.
Justification for the selection of the test system: the Brachydanio rerio was chosen as a Cyprinidae species, since it is widely accepted by Health Authorities as an experimental model with documented susceptibility to a wide range of toxic substances.
Number: 10 animals/concentration
Body weight and length (at the beginning of the study): 0. 330 g and about 2.7 cm (average values).
Supplier: F.lli Richiardi, Gaglianico (BI) - Shipping slip no. 04/A dated March 27, 1997
Water: reconstituted water with a hardness of about 236 mg/l of CaCO3, a Ca/Mg rate of 4:1 and a Na/K of 10:1.

ANIMAL HUSBANDRY
Keeping of fish
All fish are kept (in a glass aquarium) in water having the following characteristics:
- Temperature: 23˚C
- pH: about 7.8
- Hardness: about 236 mg/l of CaCO3
- Dissolved O2 concentration: 8.1 mg/l
The fish are exposed to artificial lighting with a light period of 16 hours daily and are fed once a day with Mangime SERA (Aquaristik GmbH).

Supply and acceptance of fish
The fish were purchased in excess of the number foreseen for the study. On arrival all fish were kept in a glass aquarium and following a 48-hour setting in period, no mortality was recorded after 7 days of acclimation.

Study acclimation
After acceptance selected fish are kept for a further period of 5-7 days before use, to complete the acclimation.

Identification of the experimental groups
During the experiment the fish are kept in silylated glass beakers containing 5 liters of test article solution or control. Beaker labels gave experiment number and concentration group and color tag. Each beaker was replaced by a new one each time the test article solution and control were changed.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

236 mg CaCO3/L

Test temperature:

22˚C

pH:

7 - 8.3

Dissolved oxygen:

8.1 mg/L

Salinity:

Not specified

Conductivity:

Not specified

Nominal and measured concentrations:

Nominal concentrations of 0.033, 0.104, 0.33 mg/l/day

Key result

Duration:

21 d

Dose descriptor:

LC50

Effect conc.:

> 0.33 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.33 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality

Details on results:

OBSERVATION OF VISIBLE ABNORMALITIES
No animals of the control and the treated group showed visible signs of abnormalities.

MORTALITY
No animals of the controls and the treated groups died.

BODY WEIGHT
No effects

PARAMETER CHECKING
The values of dissolved oxygen, pH and temperature always remained within the acceptable limits.

LC50 AND ITS STATISTICAL LIMITS, LOEC AND NOEC
LC50 was not calculable at any time.
LOEC: was not calculable
NOEC: for lethal and other statistically significant effects was 0. 33 mg/l.

Comments: Concentration analyses of the test solutions were performed and samples withdrawn at 0 and 24 h of the first and the last day of treatment with the HPLC method (see 1.4.10). The test substance disappeared, therefore all calculations are based on nominal concentrations. A later stability test on aqueous solutions of Anox BF stated that the most reasonable explanation for the loss of substance in aqueous solution is the hydrolysis of the esteric function. This hydrolysis process would lead to the formation of a mixture of the C-14 and C-15 alcohols and 3-(3'-5' di t- butyl-4' hydroxylphenyl)propionic acid. However due to the low concentrations of these decomposition products it has not been possible to confirm their presence in solution.

Validity criteria fulfilled:

yes

Conclusions:

Experimental data from a prolonged toxicity study in which fish were treated with the test article ANOX BF at the nominal concentrations of 0, 0.033, 0.104 and 0.33 mg/l/day.
The animals were treated and observed for 21 (1 4+7) days.
No animals in the control and in the treated groups died or showed signs of visible abnormalities.
The LC50 value was not calculable at any time.
LOEC was not calculable at any time.
NOEC for lethal and other than lethal effects was always 0.33 mg/l (nominal concentration).

Executive summary:

The purpose of this study was to determine at selected intervals the threshold levels of lethal and other observable effects and the NOEC in fish after prolonged exposure to the test article ANOX BF.

The test method is in accordance with Organisation for Economic Cooperation and Development Guidelines (section 2, subpart 204, Paris 1984).

Experimental data from a prolonged toxicity study in which fish were treated with the test article ANOX BF at the nominal concentrations of 0, 0.033, 0.104 and 0.33 mg/l/day.
The animals were treated and observed for 21 (1 4+7) days.
No animals in the control and in the treated groups died or showed signs of visible abnormalities.
The LC50 value was not calculable at any time.
LOEC was not calculable at any time.
NOEC for lethal and other than lethal effects was always 0.33 mg/l (nominal concentration).

Description of key information

The LC50 value and LOEC was not calculable at any time. NOEC for lethal and other than lethal effects was 0.33 mg/l (nominal concentration).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The purpose of this study was to determine at selected intervals the threshold levels of lethal and other observable effects and the NOEC in fish after prolonged exposure to the test article ANOX BF.

The test method is in accordance with Organisation for Economic Cooperation and Development Guidelines (section 2, subpart 204, Paris 1984).

Experimental data from a prolonged toxicity study in which fish were treated with the test article ANOX BF at the nominal concentrations of 0, 0.033, 0.104 and 0.33 mg/l/day.
The animals were treated and observed for 21 (1 4+7) days.
No animals in the control and in the treated groups died or showed signs of visible abnormalities.
The LC50 value was not calculable at any time.
LOEC was not calculable at any time.
NOEC for lethal and other than lethal effects was always 0.33 mg/l (nominal concentration).