Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Qualifier:
according to
Guideline:
EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Test item name: Fatty acid, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine
Batch number : TEE3316/22
Molecular weight: > 700
Declared purity: 97 %
ChemService sample code: 2901348-001
Storage conditions: Room temperature/dark conditions

Sampling and analysis

Analytical monitoring:
yes

Test solutions

Vehicle:
no

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
Juvenile fish were used and all of them were approximately the same age and size. In accordance with the test guidelines, the fish were held in the ChemService laboratories for more than 12 days at the same conditions regarding water quality, temperature and illumination, as during the test. During the holding period (21 days), the fish were fed with commercial fish food (Tetramin Granules) until 48 hours before the test initiation.
The used batch of fishes was accepted since no mortality and no signs of distress were observed during the acclimation period.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h

Test conditions

Hardness:
10 - 250 mg/L as CaCO3
Test temperature:
The measured values ranged between 20.4 and 23.9 °C.
pH:
7.14 – 7.34
Dissolved oxygen:
During the test period, the dissolved oxygen concentrations in the test medium and the control were always above 60 per cent of the air saturation value .
Details on test conditions:
Photoperiod: 16 hours light and 8 hours darkness daily ; light intensity was 50 - 500 lux, with 2 daily 30-minutes transition periods

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality

Any other information on results incl. tables

A limit test under static conditions was performed to evaluate the effect of the test item on zebra fish (Danio rerio).

The fishes were checked for mortality and abnormalities in appearance and behaviour after 2, 24, 48, 72 and 96 hours.

In the control 0% mortality was observed during the test period. This value complies with the validity criteria of the test, according to the mentioned guidelines, that provide a maximum mortality of fishes exposed of 10% in the control medium at the end of the test.

 

The observed effects in the test concentration and in the control are reported in Table 1.

Table 1           

Lethal effect of Fatty acid, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine to zebra fish.

Nominal

test item concentration

[mg/L]

Replicate

n. of exposed

fish

n. dead fish

2 h

24 h

48 h

72 h

96 h

0.0

(Negative control)

A

10

0

0

0

0

0

0.0

(Negative control)

B

10

0

0

0

0

0

100.0

A

10

0

0

0

0

0

100.0

B

10

0

0

1

1

1

The NOEC and the LOEC were determined directly from the Raw Data by statistical comparison with the control (Fisher’s Exact test) at each observation time.

 

The NOEC (highest concentration tested without toxic effects after the exposure period) of test item to zebra fish was determined to be equal to the tested concentration, 100.0 mg/L.

Based on the observed effects, the LOEC and the 96-h LC50were determined to be greater than 100.0 mg/L.

As for the test medium appearance, no remarkable observation was made. The test medium was a clear solution throughout the whole test duration.

Light intensity during the test was measured at test initiation, after 48 and 96 hours and it was in the range 208 – 272 Lux, according to the mentioned guidelines (50 – 500 Lux).

The range of temperature in the control tanks was between 20.8 –with a mean value ofand a standard deviation of 0.9; while the temperature values in the test item solutions ranged between 20.4 andwith a mean value ofand a standard deviation of 0.8. The temperature values measured were slightly below the recommended range, however this deviation did not affect the outcome of the study.

During the test period, the dissolved oxygen concentrations in the test medium and the control were always above 60 per cent of the air saturation value as showed in Table 2.

The pH values were in the range 7.03 - 7.45 as required by the mentioned guidelines and did not vary more than 0.11 units (Table 2) between the start and the end of test period, thus meeting the validity criteria for pH (maximum difference = 1.5 pH unit). 

Table 2.          

Dissolved oxygen concentrations and pH values in the test media at test initiation and during the exposure time.

Nominal test item concentration (mg/L)

 

 

 

 

 

(Negative control)

A

 (Negative control)

B

100.0

A

100.0

B

 

Dissolved

 

oxygen

 

(%)

 

 

 

0 h

64.5

63.8

63.3

64.9

24 h

77.7

77.1

70.9

72.6

48 h

73.4

72.6

70.2

74.5

72 h

74.5

73.8

72.7

73.1

96 h

76.7

73.7

73.3

71.9

 

 

 

pH

 

 

 

 

 

 

0 h

7.14

7.19

7.28

7.34

24 h

7.00

7.13

7.24

7.28

48 h

7.31

7.42

7.45

7.42

72 h

7.28

7.32

7.38

7.36

96 h

7.03

7.23

7.30

7.32

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The test item “Fatty acid, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine” caused 0.5% mortality to zebra fish after 96-hours of exposure at the nominal product concentration of 100 mg/L in a static test.
The NOEC, LOEC and LC50 values after 96 h, calculated based on the nominal concentration, were assessed to be as follows:

96 h NOEC: 100.0 mg/L
96 h LOEC: > 100.0 mg/L
96 h LC50: > 100.0 mg/L
96 h LC50: > 100.0 mg/L

Executive summary:

The acute toxicity of the test item Fatty acid, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine to zebra fish (Danio rerio)was determined in a limit, 96‑hour static test according to:

- OECD Guideline for Testing of Chemicals, N. 203. “Fish, acute toxicity test”, 1992

- OPPTS 850.1075 guideline “Fish acute toxicity test, freshwater and marine”, 1996.

For this purpose, juvenile fish of the speciesDanio reriowere exposed, after an acclimation period (21 days), to an aqueous test medium containing the test item at the only nominal concentration of 100.0 mg/L of product. Two replicate for the test concentration and for negative control were prepared.

The test organisms were checked for mortality after 2, 24, 48, 72 and 96 hours from test initiation.

 

The pH values of the test medium after preparation was in the range 7.28 – 7.34 for test item and in the range 7.14 – 7.19 for negative control. Therefore, the pH of the test medium was not adjusted before the start of the test.

 

The dissolved oxygen was all the time above 60 per cent of the air saturation value both in each replicate of negative control and test solution.

 

The temperature during the test was recorded by the mean of PC system and it was in the range 20.7 -for controls replicate and 20.4 -for the test item solutions. The temperature was slightly below the OECD’s recommended range (23.0±2.0 °C), however it did not affect the results of the study.

 

Light intensity during the test was measured at test initiation, after 48 and 96 hours and it was in the range 208 – 272 Lux according to the mentioned guidelines that provided a range of light intensity of 50 – 500 Lux.

 

In the control 0% mortality was observed during the test period. This value complies with the validity criteria of the test, according to the mentioned guidelines, that provide a maximum mortality of fishes exposed of 10% in the control medium at the end of the test.

 

In the test item solution one fish was found to be dead at the end of the test period, therefore the NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours) of Fatty acid, tall-oil, reaction products with diethylenetriamine, maleic anhydride, tetraethylenepentamine and triethylenetetramine to zebra fish was determined to be equal to the tested concentration (nominal 100 mg/L).

Based on the observed effects, the LOEC and the 96-h LC50were determined to be greater than the 100.0 mg/L as product.

 

During the test period any behaviour and/or pathological signs on exposed fishes were observed.