Reaction mass of 5-[{4-[Benzyl(methyl)amino]phenyl}diazenyl]-1,4-dimethyl-4H-1,2,4-triazol-1-ium bromide and 3-[{4-[Benzyl(methyl)amino]phenyl}diazenyl]-1,4-dimethyl-4H-1,2,4-triazol-1-ium bromide

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:

Study initiation date - 10 February 1994; Experiment start date - 16 February 1994; Experiment end date - 18 February 1994; Study completion date - 18 May 1994

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

Specific details on test material used for the study:

- Code number: FAT-31016/P
- EN-No.: 414154.32
- Purity: 95 %
- Appearance: solid
- Solubility: 40 g/L (in water)
- Storage: room temperature
- Expiration date: 07/98

Analytical monitoring:

yes

Details on sampling:

Sampling for analysis.
All test medium samples for the analytical determinations were taken after mixing of the test solutions from the approximate center of the test vessels and kept at -18 °C to -25 °C until analyses.
The concentrations of the test substance at the beginning and at the end of the test was analysed by the Analytical Services K, Textile Dyes Division.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- The test water medium was prepared with bidistilled water.
- Stock solution: 0.5 g test substance were mixed with 200 ml test medium and fill up to 500 ml with test medium. The stock solution was homogenized for 5 minutes in the ultrasonic bath.
- The test medium used had the following composition:
NaHCO3 65 mg/l
MgSO4 * 7 H2O 123.3 mg/l
KCl 5.8 mg/l
CaCI2 * 2 H2O 294 mg/l
Na2SiO3*9H2O 10 mg/l
NaNO3 274 µg/l
KH2PO4 143 µg/l
K2HPO4 184 µg/l

- The pH was in a range of 7.8 - 8.2.
- The medium was oxygen saturated at the beginning of the test.
- Theoretical water hardness: 136 mg/l CaCO3
- Ca / Mg ratio: 4:1
- Na / K ratio: 10:1
- alkalinity: 0.8 mmol/l

Test organisms (species):

Daphnia magna

Details on test organisms:

The Daphnia were bred in the testing facility of the Product Ecology TF, Textile Dyes Division under standardised conditions.

Breeding:
Cultures of Daphnia magna are maintained in glass vessels containing approx.. 3.0 litre of reconstituted water (test medium) at 18 - 22 °C. The water was renewed partially three times weekly. The daphnids were feed with a suspension of green algae (Scenedesmus subspicatus) supplemented by a suspension of Tetramin-extract in such quantities that the food was consumed within 24h.

Pretreatment:
24 hours before the start of the exposure reproductive daphnids were separated from the young daphnids by sieving all individuals through a 710 µm sieve. The quality of the daphnids is checked at least every 3 months by determining the EC50 values (24 h) for potassium dichromate (range: 0.8 -1.5 mg/l). For each concentration, of test substance, 20 daphnids divided in two groups of 10 animals are used.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

Theoretical water hardness: 136 mg/l CaCO3

Test temperature:

21 ± 1°C

pH:

7.7 - 8.2.

Dissolved oxygen:

92 - 99 %

Nominal and measured concentrations:

Based on a pretest, the test concentrations were chosen as 0.58, 1.0, 1.8, 3.2, 5.8 and 10.0 mg test article / I nominal.

Details on test conditions:

48 hours at 21 °C, 16 hours light / 8 hours dark
The Daphnia were exposed to the test article in 250 ml beakers containing 100 - 200 ml test medium with 10 animals per beaker. All tests were run in duplicate. During exposure the animals were kept at 21 ± 1 °C with 16 hours illumination.

- Determination of results
Test evaluations were done according to the guidelines by visual assessment of mobility of the Daphnia after 24 and 48 hours.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

3.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

In control and test concentrations 0.5 and 1.0 mg/l , no immobilisation was observed. At 1.8 mg/l, 80 % immobility (9/10) was observed at the end of 48 hour observation period. At test concentrations 3.3, 5.8 and 10.0 mg/l, 100 % immobility (10/10) was observed.

Results with reference substance (positive control):

Potassium dichromate (EC50: 1.01 mg/l)

Reported statistics and error estimates:

The EC50 - value are calculated with the " Spearman - Kaerber's " method.

The reported results are related to the analytical concentrations of the test article. During test duration the test article concentrations in the analysed test media were in the range of 80 - 120 % of the nominal values. Therefore, under test conditions the test article was sufficiently stable.

Validity criteria fulfilled:

yes

Conclusions:

The EC50 value was determined to be 1.5 mg/l.

Executive summary:

The acute toxicity of FAT - 31016/P to Daphnia magna was determined in a 48-hours static test according to the OECD Guideline 202 and EEC Directive C 2. The test was performed in compliance with the Good Laboratory Practice Regulations. The nominal concentrations tested were 0.58, 1.0, 1.8, 3.2, 5.8 and 10.0 mg test article / l, and a control without any additions. The reported results are related to the analytical concentrations of the test article. During test duration the test article concentrations in the analysed test media were in the range of 80 - 120 % of the nominal values. Therefore, under test conditions the test article was sufficiently stable. In all test concentrations non alteration of the test substance after 48 hours was observed. In the control non immobilization of the Daphnia magna was observed after 48 hours. At test concentrations 0.5 and 1.0 mg/l, no immobilisation was observed, while at 1.8 mg/l, 80 % immobility was observed at the end of 48 hour observation period. At test concentrations 3.3, 5.8 and 10.0 mg/l, 100 % immobility was observed. Hence, the EC0 value was at least 1.0 mg/l, but might even be higher than this concentration. The EC50 value was determined to be 1.5 mg/l, while the EC100 value was 3.3 mg/l after 48 hours.

Description of key information

The EC50 value to Daphnia magna was determined to be 1.5 mg/l, while the EC100 value was 3.3 mg/l after 48 hours.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1.5 mg/L

Additional information

The acute toxicity of FAT - 31016/P to Daphnia magna was determined in a 48-hours static test according to the OECD Guideline 202 and EEC Directive C 2. The test was performed in compliance with the Good Laboratory Practice Regulations. The nominal concentrations tested were 0.58, 1.0, 1.8, 3.2, 5.8 and 10.0 mg test article / l, and a control without any additions. The reported results are related to the analytical concentrations of the test article. During test duration the test article concentrations in the analysed test media were in the range of 80 - 120 % of the nominal values. Therefore, under test conditions the test article was sufficiently stable. In all test concentrations non alteration of the test substance after 48 hours was observed. In the control non immobilization of the Daphnia magna was observed after 48 hours. At test concentrations 0.5 and 1.0 mg/l, no immobilisation was observed, while at 1.8 mg/l, 80 % immobility was observed at the end of 48-hour observation period. At test concentrations 3.3, 5.8 and 10.0 mg/l, 100 % immobility was observed. Hence, the EC0 value was at least 1.0 mg/l, but might even be higher than this concentration. The EC50 value was determined to be 1.5 mg/l, while the EC100 value was 3.3 mg/l after 48 hours.