Reaction mass of Fatty acids, montan-wax and Fatty acids, montan-wax, calcium salts and Montan wax

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

From FEB 13 to FEB 27 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to relevant guideline and compliant to GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

As the substance was practically insoluble in water, instead of 5 concentrations as recommended by the guideline only 4 concentrations were tested.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Montan waxes, type WE 4

IUPAC Name:

Montan waxes, type WE 4

Constituent 2

Reference substance name:

Reaction mass of Fatty acids, montan-wax and Glycerides, montan-wax and Montan wax

EC Number:

914-469-2

Molecular formula:

not applicable

IUPAC Name:

Reaction mass of Fatty acids, montan-wax and Glycerides, montan-wax and Montan wax

Details on test material:

"Licolub WE 4", ester of montanacids;
active content: 100 %

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

Not applicable

Test solutions

Vehicle:

no

Details on test solutions:

According to "Guidance Document On Aquatic Toxicity Testing Of Difficult Substances And Mixtures" of the OECD, Environmental Health and Safety Publications, No. 23 - 3.1 Poorly water-soluble substances.

A definite amount of the test substance (10 mg; 100 mg; 1 g etc.) was filled into measuring flask and the total volume made up with water at 21 +/- 1 deg C.
The solution was shaken during 24 hours. The test substance was not soluble in water. Other solvents couldn´t be used because of its toxic properties.
Shaking produced a dispersion of the test substance. Then the solution was filtrated. The hydrophobic properties of "Licolub WE 4" didn't allow an investigation of higher concentrations (more than 10 g/L).
The test bottles were conditioned about 15 minutes before the test was started.
The pH value and oxygen concentration were measured at the beginning and the end of the test.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Brachydanio rerio
Average length of the fishes: 2.0 cm;
Feeding: daily, day/night change, circulation filters;
Temperature of water approximately 21 deg C;
A minimum of 14 days the fishes were kept under these conditions before test.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

Dilution water: ISO 6341-1982 as specified in Annex II of OECD 203 (25 ml each of solutions CaCl2, MgSO4, Na2CO3 and KCI were mixed and the total volume made up to 1 litre with deionised water.

Test temperature:

The temperature conditions were kept constant at 21 +-1 deg C

pH:

For details, see table section "Any other information on results incl. tables"
Control
test start: 7.8; test stop: 7.3;

Treatment groups:
test start: 7.3 (11.8 mg/L) - 7.7 (1000 mg/L); test stop: 7.4 (10000 mg/L) - 7.9 (1000 mg/L);

Dissolved oxygen:

For details, see table section "Any other information on results incl. tables"
Control
test start: 6.5 mg/L; test stop: 6.4 mg/L;

Treatment groups:
test start: 6.2 mg/L (11.8 mg/L test item) - 6.5 mg/L (100 mg/L test item); test stop: 5.9 mg/L (10000 mg/L test item) - 6.4 mg/L (100 mg/L test item);

Salinity:

Dilution water: ISO 6341-1982 as specified in Annex II of OECD 203 (25 ml each of solutions CaCl2, MgSO4, Na2CO3 and KCI were mixed and the total volume made up to 1 litre with deionised water.

Nominal and measured concentrations:

The following nominal concentrations of the submission substance were applied for the treatment groups:
[mg/L] 11.8, 100, 1000, 10000.
The test substance was not soluble in water. Therefore, no concentration measurements were performed with the filtrated solutions. Nominal concentrations were applied and filtered solutions were used in the test

Details on test conditions:

The determination of the acute toxicity was performed as described in the OECD 203 method. The species Branchydanio rerio was used as test fish according to OECD 203.
These fishes were put into the test solution of chosen concentrations. 7 fishes were used for each test concentration at a test volume of 7 litre. The fishes were inspected after 24, 48, 72 and 96 hours. The temperature conditions were kept constant at 21 +/-1 deg C. The test vessels were exposed to indirect daysimilar light (light-dark cycle for 12 hours).

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

10 g/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 g/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

During the test period (96 hours) no toxic effects of the fishes were observed. All tested animals were still alive. No mortalities occurred with the control.

Reported statistics and error estimates:

A Probit analysis was not possible as no effect was observed.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Details on measurements:

Test item concentration	pH 0 h	pH 96 h	O2 [mg/L] 0 h	O2 [mg/L] 96 h
control solution	7.8	7.3	6.5	6.4
11.8	7.3	7.6	6.2	6.1
100	7.4	7.7	6.5	6.4
1000	7.7	7.9	6.3	6.2
10000	7.6	7.4	6.4	5.9

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In an acute toxicity test with the submission substance on Brachydanio rerio according to OECD 203 the following effect values were determined:
NOEC (96h, mortality and behaviour) = 10 g/L (nominal); LC50 (96h) > 10 g/L (nominal).

Executive summary:

In an acute toxicity test with the submission substance on Brachydanio rerio according to OECD 203 and compliant to GLP (reliability catogory 1), no effects neither regarding mortality nor behavioural abnormalities were observed. Due to the very poor solubility of the test item in water, nominal concentrations were applied and filtered solutions used in the test. The following effect values were determined:

NOEC (96h, mortality and behaviour) = 10 g/L

LC50 (96h) > 10 g/L.