Reaction mass of benzyl 2-ethylhexyl adipate and bis(2-ethylhexyl) adipate and dibenzyl adipate

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:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Aliquots of the samples from the biological test were directly analysed by HPLC and UV/VIS-detection (range of the injection volume: 100 μL, depending on the expected concentration).
The test item consists of three components. Only two components can be detected.

Details on test solutions:

Reconstituted water prepared according to the recommendations of ISO 7346. This freshly prepared standard dilution water was used for the maintenance of the test animals under flow-through conditions and for the preparation of stock and test solutions of the test item.
The total hardness of the dilution water, measured at test start, was 13.5 °dH (= 241 mg/L CaCO3).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

- Source : Aqua KlöGer (Germany)
- Date supplied : 2011-10-26
- Acclimatisation : Stock held since 2011-10-26 and acclimatised to the test conditions since then.
- Temperature : 20-24 °C
- Dissolved oxygen : > 5 mg/L
- Feeding : Commercial fish food, daily. Feeding discontinued 24 h prior to test start.
- Mortalities during acclimatisation period : < 5 %
- Medication : none
- Mean standard length (n = 14): 3.42 cm (S.D. = 0.20 cm)

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

13.5°dH (= 241 mg/L CaCO3)

Test temperature:

21.8 to 22.7°C

pH:

7.7-7.8

Dissolved oxygen:

8.7 mg/L (=98.5 % saturation)

Nominal and measured concentrations:

Nominal concentration: 100 mg/L

Details on test conditions:

Pre-treatment of test item and preparation of test item concentrations:
To produce the only Lethal Loading rate a Water Accommodated Fraction (WAF) was prepared daily for the test concentration 100 mg/L. The test item was added to dilution water and treated with ultra turrax for 60 seconds at 8000 rpm and afterwards stirred for 24 hours on a magnetic stirrer. Finally undissolved particles of the test item were removed by filtration using a folded filter of pore size 7-12 μm and an aseptic filter Sartobran 150 Sterile Capsule with a pore size 0.45 + 0.2 μm. The pH of the loading was determined and ranged between 7.7 and 7.8.
Finally 7 fish were given to all test item concentrations and the control.
Exposure conditions:
Test vessels : glass aquaria holding 5 L of test media covered by glass plates
Experimental design : 1 test concentration plus 1 control, 7 fish per test concentration, no feeding during the expo-sure period, semi static
Method of initiation : fish were placed in prepared media
Loading : 3.07 g body weight (wet weight) per litre
Photoperiod : 16 h light: 8 h dark
Temperature : 21.8 to 22.7 °C
Aeration : gentle aeration via narrow glass tubes
Test item concentration/s : 100 mg/L
Method of administration : direct weighing
Medium renewal : daily
Duration of exposure: 96 hours
Criteria of effects:
The criterion of death used in this study was the absence of response to physical stimulation. In addition to observations on mortality at 2, 24, 48, 72 and 96 hours, type and incidence of sub lethal effects compared with control fish were observed.

Duration:

2 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LL50

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:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No toxic effects against fish were observed at a limit Lethal Loading of 100 mg/L, corresponding to a measured concentration of < 0.131 mg/L, which reflects the maximum water solubility under exposure conditions.

Sublethal observations / clinical signs:

Some test fish had a somewhat larger size than the one recommended in the test guideline for the zebra fish. This deviation is not regarded to be relevant to the results.

Validity criteria fulfilled:

yes

Remarks:

Mortality in the controls < 10% by the end of the test. Dissolved oxygen concentration above 60% of the air-saturation value throughout the exposure period. The pH did not vary by more than 1 unit.

Conclusions:

The acute toxicity of 'Reaction mass of benzyl 2-ethylhexyl adipate and bis(2-ethylhexyl) adipate and dibenzyl adipate' to fish (Danio rerio) was conducted in a semi-static test according to EU Method C.1 (Acute Toxicity for Fish). No toxic effects against fish were observed at a limit Lethal Loading (LL50) of > 100 mg/L, corresponding to a measured concentration of < 0.131 mg/L, which reflects the maximum water solubility under exposure conditions.

Executive summary:

A study was performed to assess the acute toxicity of 'Reaction mass of benzyl 2-ethylhexyl adipate and bis(2-ethylhexyl) adipate and dibenzyl adipate' to Danio rerio under semi static conditions.

The study was conducted in accordance with Council Regulation (EC) No 440/2008, Method C.1 'Acute toxicity for Fish' (2008) which is in most parts equivalent to the OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992).

A group of seven fish of the recommended size was exposed to a limit test concentration of nominally 100 mg/L of 'Reaction mass of benzyl 2-ethylhexyl adipate and bis(2-ethylhexyl) adipate and dibenzyl adipate' dissolved in dilution water. Auxiliaries used to prepare the test media were an ultra turrax, a magnetic stirrer, folded filters and aseptic filters. Observations were made on the number of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure.

No toxic effects against fish were observed at a limit Lethal Loading of 100 mg/L, corresponding to a measured concentration of < 0.131 mg/L, which reflects the maximum water solubility under exposure conditions.

All results are expressed in terms of Lethal Loadings (LL). In the freshly prepared test media and in the media after 24 hours of exposure the values were below the quantification limit of the HPLC analysis (0.131 mg/L).

'Reaction mass of benzyl 2-ethylhexyl adipate and bis(2-ethylhexyl) adipate and dibenzyl adipate' consists of three components, therefore a Water Accommodated Fraction (WAF) was used to test effects at a limit lethal loading of 100 mg/L.

Description of key information

The acute toxicity of 'Reaction mass of benzyl 2-ethylhexyl adipate and bis(2-ethylhexyl) adipate and dibenzyl adipate' to fish (Danio rerio) was conducted in a semi-static test. No toxic effects against fish were observed at a limit Lethal Loading (LL50) of > 100 mg/L, corresponding to a measured concentration of < 0.131 mg/L, which reflects the maximum water solubility under exposure conditions.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

A study was performed to assess the acute toxicity of 'Reaction mass of benzyl 2-ethylhexyl adipate and bis(2-ethylhexyl) adipate and dibenzyl adipate' to Danio rerio under semi static conditions.

The study was conducted in accordance with Council Regulation (EC) No 440/2008, Method C.1 'Acute toxicity for Fish' (2008) which is in most parts equivalent to the OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992).

A group of seven fish of the recommended size was exposed to a limit test concentration of nominally 100 mg/L of 'Reaction mass of benzyl 2-ethylhexyl adipate and bis(2-ethylhexyl) adipate and dibenzyl adipate' dissolved in dilution water. Auxiliaries used to prepare the test media were an ultra turrax, a magnetic stirrer, folded filters and aseptic filters. Observations were made on the number of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. No toxic effects against fish were observed at a limit Lethal Loading of 100 mg/L, corresponding to a measured concentration of < 0.131 mg/L, which reflects the maximum water solubility under exposure conditions. All results are expressed in terms of Lethal Loadings (LL). In the freshly prepared test media and in the media after 24 hours of exposure the values were below the quantification limit of the HPLC analysis (0.131 mg/L). 'Reaction mass of benzyl 2-ethylhexyl adipate and bis(2-ethylhexyl) adipate and dibenzyl adipate' consists of three components, therefore a Water Accommodated Fraction (WAF) was used to test effects at a limit lethal loading of 100 mg/L.