Trisodium 2,4-diamino-3,5-bis-[4-(2-sulfonatoethoxy)sulfonyl)phenylazo]benzenesulfonate

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:

May 20, 1997 to July 10, 1997

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)

GLP compliance:

yes (incl. QA statement)

Remarks:

Swiss GLP

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: WP2/96
- Expiration date of the lot/batch: November 30,2004

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature, in the dark
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: for at least 48 h and approximately 50 g/1 (at 20 °C) in water

Analytical monitoring:

yes

Details on sampling:

For the analytical measurements of the real test substance concentrations, duplicate samples from the freshly prepared test medium and from the control were taken just before the start of the test and on Day 1 of exposure.
For the determination of the stability of the test substance under the test conditions respectively the maintenance of the test substance concentrations during the test medium renewal period, duplicate samples from the test medium of the single test concentration and the control were collected after 24 hours (Day 1) by pouring together the contents of the test beakers of each treatment.

Vehicle:

no

Details on test solutions:

no data

Test organisms (species):

Daphnia magna

Details on test organisms:

The study was performed with young test animals of a clone of the species Daphnia magna Straus. The clone was originally supplied by the University of Sheffield/UK in 1992, defined from the supplier as clone 5. Since this date the clone is bred in the laboratories of RCC under similar temperature and light conditions as in the test, and in reconstituted water of a similar quality regarding to pH, components of the main ions and total hardness as the test water used in the test.
At the start of the test the used test animals were 6-24 h old and were not first brood progeny.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

2.5 mmol/l (= 250.0 mg/l) as CaCO3

Test temperature:

20-21 °C

pH:

7.6-8.0

Dissolved oxygen:

8.4-8.8 mg/l

Salinity:

no data

Nominal and measured concentrations:

nominal: 300 mg/l

Details on test conditions:

In each treatment, the single test concentration and the control, 20 Daphnia were tested, divided into two groups of ten animals, each group in 50 ml test medium in a 100-ml glass beaker. The test vessels were labelled with the RCC Project number and all necessary additional information to assure unmistakable identification. The test was performed in an air-temperated room.
Light conditions: a 16-h light to 8-h darkness photoperiod, light intensity at light period between 200 and 1200 Lux
A semi-static test with test medium renewal after 24 h was chosen to keep the concentration of the test substance in the test medium as constant as possible during the test period of 48 h. After 24 h the test animals were placed in a clean test vessel with freshly prepared test medium.
Each day, the pH-values and the dissolved oxygen concentrations were determined in the freshly prepared and old test medium of the single test concentration and the control. The water temperature was determined in the test medium of one control beaker at the same dates.

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

>= 259 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 259 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 259 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 259 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Reported statistics and error estimates:

The NOEC and the EC 0 were determined directly from the raw data. The EC 50 and the EC 100 could not be quantified due to the absence of toxicity of the test substance.

A limit test was performed in accordance with the Commission Directive 92/69/EEC to demonstrate that the test substance has no toxic effect on Daphnia up to the concentration of at least 100 mg test substance/l. The analytically determined test substance concentrations in the freshly prepared test media

ranged from 99 % to 101 % of the nominal value (see analytical results, Attachment Table 2). During the renewal period of 24 hours the test substance concentration decreased to 72 % of nominal, if the quantification is based on main compound. If the quantification is based on the sum of main compound and three main degradation products, the mean recoveries in the non-aged samples were 100 % and 98 %. In the 24 h aged samples the mean recovery was 93 %.

All reported results are related to the mean measured test substance concentration of 259 mg/l (calculated as the average over the test concentrations of the main compound measured at the start of the test and after 24 h). In the control and in the test concentration of 259 mg test substance/1 no significant number of immobilized or dead test animals was observed during the test period of 48 h.

Validity criteria fulfilled:

yes

Conclusions:

The 48-h NOEC and EC50 of FAT 45’176/A to Daphnia magna were determined to be at least 259 mg test substance/l.

Executive summary:

The acute toxicity of FAT45'176/A to Daphnia magna was determined in a 48-h semi-static test according to OECD 202 and EU C.2 guideline.

A limit test was performed in accordance with the Commission Directive 92/69/EEC to demonstrate that the test substance has no toxic effect on the test animals up to a concentration of at least 100 mg test substance/l. Since the test substance is not stable in test water a concentration of 300 mg/l was tested to make sure that the real test substance concentration in test water was at least 100 mg/l. Additionally, a control was tested in parallel.

The analytically determined test substance concentrations in the freshly prepared test media ranged from 99 % to 101 %of the nominal value. During the renewal period of 24 h the test substance concentration decreased to 72 % of nominal, if the quantification is based on main compound. All reported results are related to the mean measured test substance concentration of 259 mg/l (calculated as the average over the test concentrations measured at the start of the test and after 24 h

The 48-h NOEC (highest concentration tested without toxic effects after the exposure period of 48 h), respectively the 48-h EC₀of FAT 45’176/A to Daphnia magnawere determined to be at least 259 mg test substance/l. The NOEC and EC0 might even be higher than this concentration, but concentrations in excess of 259 mg test substance/l have not been tested. The 48-h LOEC (lowest concentration with toxic effects) and the 48- h EC₅₀were clearly higher than 259 mg test substance/l.

Description of key information

The EC50value of Reactive Orange 136 after 48 h was 259 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

259 mg/L

Additional information

The acute toxicity of FAT45'176/A to Daphnia magna was determined in a 48-h semi-static test according to OECD 202 and EU C.2 guideline.

A limit test was performed in accordance with the Commission Directive 92/69/EEC to demonstrate that the test substance has no toxic effect on the test animals up to a concentration of at least 100 mg test substance/l. Since the test substance is not stable in test water a concentration of 300 mg/l was tested to make sure that the real test substance concentration in test water was at least 100 mg/l. Additionally, a control was tested in parallel.

The analytically determined test substance concentrations in the freshly prepared test media ranged from 99 % to 101 %of the nominal value. During the renewal period of 24 h the test substance concentration decreased to 72 % of nominal, if the quantification is based on main compound. All reported results are related to the mean measured test substance concentration of 259 mg/l (calculated as the average over the test concentrations measured at the start of the test and after 24 h

The 48-h NOEC (highest concentration tested without toxic effects after the exposure period of 48 h), respectively the 48-h EC₀of FAT 45’176/A to Daphnia magnawere determined to be at least 259 mg test substance/l. The NOEC and EC0 might even be higher than this concentration, but concentrations in excess of 259 mg test substance/l have not been tested. The 48-h LOEC (lowest concentration with toxic effects) and the 48- h EC₅₀were clearly higher than 259 mg test substance/l.

In another supporting study, the acute toxicity of FAT45'176/A to Daphnia magna was determined in a 48 h semi-static test according to the OECD 202 guideline. The 48-h NOEC and EC₅₀ were clearly higher than 63 mg/l.