Reaction mass of pentasodium 2-{[4-chloro-6-(ethyl{3-[(2-sulfonatoethyl)sulfonyl]phenyl}amino)-1,3,5-triazin-2-yl]amino}-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate and tetrasodium 2-[(4-chloro-6-{ethyl[3-(vinylsulfonyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 March 2006 to 18 April 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40824/A
Batch: Red ROE 805 BOP 04/05
Appearance: dark red powder
Purity: Organic part (Na-salt): approx. 82 %; Main component 1 : approx. 36.2 %; Main component 2: approx. 27.5 %; Oligomers: 10%
Expiration date: 01 October 2010
Stability in water: Max. 7 days at room temperature
Solubility in water: >100g/Lat20°C
Storage: At room temperature at about 20 °C, in a desiccator because test substance is hygroscopic, away from direct sunlight

Analytical monitoring:

yes

Details on sampling:

Duplicate samples from each test medium and the control (without daphnias) just before the start of the test.
Duplicate samples from each test medium and the control after 96 hours.
All samples were taken from the approximate center of the aquaria without mixing of the test media, and were deep-frozen (at about -20 °C) immediately after sampling.

Vehicle:

no

Details on test solutions:

The following nominal concentrations were tested: 4.6, 10, 22, 46 and 100 mg/L. Additionally, a control was tested in parallel.
The test medium of the highest nominal concentration of 100 mg/L was prepared by dissolving 50.0 mg of test item completely in 500 mL of test water using intense stirring for 15 minutes at room temperature. Adequate volumes of this test medium were diluted with test water to prepare the test media with the lower test item concentrations.
The test media were prepared just before introduction of the daphnids.

Test organisms (species):

Daphnia magna

Details on test organisms:

The study was performed with young daphnids of a clone of the species Daphnia magna Straus.
At the start of the test, the organisms used in the test were 6-24 hours old and were not first brood progeny.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

250 mg/L as CaCO3

Test temperature:

20 °C

pH:

7.7 - 7.8

Dissolved oxygen:

8.6 - 8.8 mg/L

Salinity:

No data

Nominal and measured concentrations:

Nominal: 4.6, 10, 22, 46 and 100 mg/L.
Measured: the nominal test item concentrations of 46 and 100mg/L were between 90 and 92 % of the nominal values at the start and the end of the test.

Details on test conditions:

The test was performed in 100 mL glass beakers filled with 50 mL of test medium.
The beakers were covered with glass plates to reduce the loss of water and to avoid the entry of dust into the solutions.
At each test concentration and for the control, 20 daphnids were used divided into four replicates of five daphnids each.
Light conditions: A 16-hour light to 8-hour dark photopehod (with a 30 minute transition period). Light intensity during the light period was between
approximately 470 and 640 Lux.
No feeding.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

46 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

46 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

46 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No immobility of the test organisms was observed in the control and up to and including the test item concentration of 46 mg/L after 24 and 48 hours.
At the highest test concentration of 100 mg/L, the immobilization rate was 25 % and 50 % respectively after 24 and 48 hours.

Results with reference substance (positive control):

The latest result of the positive control test in September 2005 (48-hour EC50: 0.81 mg/L, RCC Study No. A28102) showed that the toxic performance was valid and within the historical range of the RCC laboratory (from 1996 to 2005: 48-hour EC50: 0.53-1.1 mg/L).

Reported statistics and error estimates:

The 24-hour EC50 of the test item could not be calculated because the immobility rate was below 50 % up to the highest test concentration after 24 hours of exposure.
The 48-hour EC50 was determined directly from the raw data (the 95 % confidence limits could not be determined). The NOEC, EC0, and EC100 were also determined directly from the raw data.

During the first 24 hours of the test, no immobility of the test organisms was observed in the control and up to and including the test item concentration of 46 mg/L. At the highest test concentration of 100 mg/L, the immobilization rate was 25% after 24 hours.

The 24-hour EC0 was 46 mg/L. The 24-hour EC50 and EC100 of the test item and their 95% confidence limits could not be determined due to the low toxicity of the test item. However, these values were clearly above the test item concentration of 100 mg/L.

After 48 hours of exposure, no immobility of the test organisms was observed in the control and up to and including the test item concentration of 46 mg/L. At the highest test concentration of 100 mg/L, the immobilization rate was 50% at the end of the test after 48 hours.

The 48-hour EC50 was determined to be 100 mg/L as the immobilization rate was exactly 50% at the test item concentration of 100 mg/L (the 95% confidence limits could not be determined). The 48-hour ECO and the 48-hour NOEC (highest concentration tested without toxic effects after 48 hours) of FAT 40824/A were both 46 mg/L since no immobilization was observed in the test organisms exposed to the test item at levels of up to and including this test concentration. The 48-hour EC100 could not be determined due to the low toxicity of the test item. However, the 48-hour EC100 was clearly above 100 mg/L. Concentrations above 100 mg/L were not tested in accordance with the test guidelines.

No remarkable observations were made concerning the appearance of the test media. All test media were clear solutions colored by the test item throughout the entire test duration.

At the beginning and the end of the test period, the dissolved oxygen concentrations in the test media and the control were at least 8.6 mg/L. The pH values of the test media and the control were between 7.7 and 7.8 and the water temperature was 20 °C at the start and the end of the test.

Validity criteria fulfilled:

yes

Conclusions:

The 24-hour EC0 was 46 mg/L while EC50 and EC100 were above 100 mg/L. The 48-hour EC50 was determined to be 100 mg/L while EC0 and were both 46 mg/L. The 48-hour EC100 were above 100 mg/L.

Executive summary:

The acute toxicity of the test item FAT 40824/A to Daphnia magna was determined in a 48-hour static test according to the EU Commission Directive 92/69/EEC, Part C.2 (1992), and the OECD Guideline for Testing of Chemicals, No. 202 (2004) The nominal test item concentrations tested were 4.6, 10, 22, 46 and 100 mg/L. The test item FAT 40824/A was stable during the test period of 48 hours under the conditions of the test. The reported biological results were based on the nominal concentrations of the test item.

During the first 24 hours of the test, no immobility of the test organisms was observed in the control and up to and including the test item concentration of 46 mg/L. At the highest test concentration of 100 mg/L, the immobilization rate was 25% after 24 hours. The 24-hour EC0 was 46 mg/L. The 24-hour EC50 and EC100 of the test item and their 95% confidence limits could not be determined due to the low toxicity of the test item. However, these values were clearly above the test item concentration of 100 mg/L. After 48 hours of exposure, no immobility of the test organisms was observed in the control and up to and including the test item concentration of 46 mg/L. At the highest test concentration of 100 mg/L, the immobilization rate was 50% at the end of the test after 48 hours.

The 48-hour EC50 was determined to be 100 mg/L as the immobilization rate was exactly 50% at the test item concentration of 100 mg/L (the 95% confidence limits could not be determined). The 48-hour ECO and the 48-hour NOEC (highest concentration tested without toxic effects after 48 hours) of FAT 40824/A were both 46 mg/L since no immobilization was observed in the test organisms exposed to the test item at levels of up to and including this test concentration. The 48-hour EC100 could not be determined due to the low toxicity of the test item. However, the 48-hour EC100 was clearly above 100 mg/L.

Based on the test results, the 24-hour EC0 was 46 mg/L while EC50 and EC100 were above 100 mg/L. The 48-hour EC50 was determined to be 100 mg/L while EC0 and were both 46 mg/L. The 48-hour EC100 were above  100 mg/L.

Description of key information

The 24-hour EC₀ was 46 mg/L while EC₅₀ and EC₁₀₀ were above 100 mg/L. The 48-hour EC₅₀ was determined to be 100 mg/L while EC₀ and were both 46 mg/L. The 48-hour EC₁₀₀ were above 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

A GLP compliant acute toxicity of FAT 40824/A to Daphnia magna was determined in a 48-hour static test according to the EU Commission Directive 92/69/EEC, Part C.2 (1992), and the OECD Guideline for Testing of Chemicals, No. 202 (2004) The nominal test item concentrations tested were 4.6, 10, 22, 46 and 100 mg/L. The test item FAT 40824/A was stable during the test period of 48 hours under the conditions of the test. The reported biological results were based on the nominal concentrations of the test item.

During the first 24 hours of the test, no immobility of the test organisms was observed in the control and up to and including the test item concentration of 46 mg/L. At the highest test concentration of 100 mg/L, the immobilization rate was 25% after 24 hours. The 24-hour EC0 was 46 mg/L. The 24-hour EC50 and EC100 of the test item and their 95% confidence limits could not be determined due to the low toxicity of the test item. However, these values were clearly above the test item concentration of 100 mg/L. After 48 hours of exposure, no immobility of the test organisms was observed in the control and up to and including the test item concentration of 46 mg/L. At the highest test concentration of 100 mg/L, the immobilization rate was 50% at the end of the test after 48 hours.

The 48-hour EC50 was determined to be 100 mg/L as the immobilization rate was exactly 50% at the test item concentration of 100 mg/L (the 95% confidence limits could not be determined). The 48-hour EC0 and the 48-hour NOEC (highest concentration tested without toxic effects after 48 hours) of FAT 40824/A were both 46 mg/L since no immobilization was observed in the test organisms exposed to the test item at levels of up to and including this test concentration. The 48-hour EC100 could not be determined due to the low toxicity of the test item. However, the 48-hour EC100 was clearly above 100 mg/L.

 

Based on the test results, the 24-hour EC0 was 46 mg/L while EC50 and EC100 were above 100 mg/L. The 48-hour EC50 was determined to be 100 mg/L while EC0 and were both 46 mg/L. The 48-hour EC100 were above  100 mg/L.