Disodium [4-hydroxy-3-[(2-hydroxy-4-nitrophenyl)azo]naphthalene-1-sulphonato(3-)][1-[(2-hydroxy-4-nitrophenyl)azo]-2-naphtholato(2-)]chromate(2-)

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:

1995

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: FAT20'011/D
Batch-Nr: 61
Purity: ca. 52.5 %
Appearance: solid
Solubility: miscible
Storage: room temperature
Expiration date: 07/99

Analytical monitoring:

yes

Details on sampling:

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

Vehicle:

yes

Details on test solutions:

Stock solution:
0.5 g test substance were mixed with 200 ml test medium in a beaker, was homogenized for 10 minutes by ultrasonic treatment and filled up to 500 ml with test medium.

Test concentrations:
The nominal test concentrations were chosen as 9.4, 21, 45, 100 and 220 mg test article/I. All test concentrations contained the vehicle Tween 80 in a concentration of 5 ppm.

In the nominal test concentrations of 9.4 - 45 mg/l after 48h no immobility of the Daphnia was observed. Therefore, the NOEC was determined to be 45 mg/l. The test concentrations below the NOEC (9.4 - 2 1 mg/l) have not been analyzed and were not used for the calculation of the EC50.

Test organisms (species):

Daphnia magna

Details on test organisms:

Test system / Test organism:
The organism used in this test system is the species Daphnia magna. The Daphnia were bred in the laboratories of the Product Ecology TF, Textile Dyes Division under standardized conditions.

Breeding:
- Cultures of Daphnia magna was held in glass vessels containing approx. 3.0 liter of reconstituted water at 18 - 22°C. The water was renewed partially three times per week. The Daphnia were fed with a suspension of green algae {Scenedesmus subspicatus) supplemented by a TETRAMIN-extract in such quantities that the food was consumed within 24h.

Pretreatment:
- 24 hours before the start of the exposure reproductive Daphnia were separated from the young Daphnia by sieving all individuals through a
1000 micrometer sieve. For each test concentration, 20 Daphnia divided in two groups of 10 animals were used.

Experimental conditions
48 hours at 21±1°C, 12 hours light/12 hours dark The Daphnia were exposed to the test substance in 250 ml beakers containing 100-200 ml test medium with 10 animals per beaker. All tests are running in duplicate.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

214 mg/l CaCO3

Test temperature:

21+/-1 °C

pH:

7.8 - 8.2

Salinity:

fresh water used

Nominal and measured concentrations:

9.4, 21, 45, 100 and 200 mg test articel/l.

- Measured concentrations:
*Start / Blank / 25.10.1994 / Content of test article in daphnia water: < 0.6 mg/l
*Ende / Blank / 27.10.1994 / Content of test article in daphnia water: < 0.6 mg/l

*Start / Vehicle / 25.10.1994 / Content of test article in daphnia water: < 0.6 mg/l
*Ende / Vehicle / 27.10.1994 / Content of test article in daphnia water: < 0.6 mg/l

*Start / 45 mg/l / 25.10.1994 / Content of test article in daphnia water: 39 mg/l
*Ende / 45 mg/l / 27.10.1994 / Content of test article in daphnia water: 39 mg/l

*Start / 100 mg/l / 25.10.1994 / Content of test article in daphnia water: 87 mg/l
*Ende / 100 mg/l / 27.10.1994 / Content of test article in daphnia water: 91 mg/l

*Start / 220 mg/l / 25.10.1994 / Content of test article in daphnia water: 197 mg/l
*Ende / 220 mg/l / 27.10.1994 / Content of test article in daphnia water: 208 mg/l
Detection limit: 0.6 mg/l (related to the nominal concentration of FAT 20011/D)

Details on test conditions:

The Daphnia were exposed to the test substance in 250 mL beakers containing 100 - 200 mL test medium with 10 animals per beaker. All tests are running in duplicate.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

39 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

146 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % confidence limit after 48 hours: 120-181 mg/l

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 203 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The test concentrations were prepared from a 1 g/L stock solution. The nominal concentrations tested were 45, 100 and 220 mg test article/I, a blank and a vehicle control without any additions. In all test concentrations non-alteration of the test substance in the test vessel was observed after 48 hours.

During the test the test article concentrations in the analysed test media were in the range of 87 - 95 % of the nominal values. Therefore, the reported results are related to the actual mean values of the analysed concentrations in the test media, measured at the beginning and at the end of the test.

No immobilization and abnormal behaviour of the Daphnia was observed in the controls. The NOEC and EC0 value after 48 hours were determined to be 39 mg/l, the EC100 value are > 203 mg/l.

Results with reference substance (positive control):

The quality of the Daphnia will be checked at least every 3 months by determining the EC50 value (24 h) for potassium dichromate (range: 0.8 -1.5 mg/l). Last reference control at September 09,1994; EC50: 1.16 mg/l.

Reported statistics and error estimates:

The EC50 values were calculated with the "Probit Model" Method (*).
(*)with the "Programme for the Computation of the ED50 "; revised November, 1988 ( V.1.2.), Ciba-Geigy AG ; W.Seewald, Mathematical Applications / FO 5.31

Validity criteria fulfilled:

yes

Conclusions:

The EC50 (48 hrs) value was found to be 146 mg/l.

Executive summary:

FAT 20011/D was tested for its acute toxicity to Daphnia magna in a 48 hours static test. The study was performed under Good Laboratory Practice (GLP) conditions in Switzerland. The experiment was done according to the OECD Guideline for Testing of Chemicals, Section 2, No. 202, adopted 4th April 1984, (issued 1993) and EU-Guideline 92/69, EEC Nr. L 383 A, C. 2, (29th Dec. 1992).

The nominal concentrations tested were 45,100 and 220 mg test article/l, a vehicle and a blank control without any additions. The test concentrations were prepared from a 1 g/L stock solution. In all test concentrations non-alteration of the test substance was observed after 48 hours.

During the test the test article concentrations in the analyzed test media were in the range of 87 - 95 % of the nominal values. Therefore, the reported results are related to the actual mean values of the analyzed concentrations of the test article in the test medium, measured at the beginning and at the end of the test.

No immobilization and abnormal behavior of the Daphnia was observed in the controls. The NOEC and EC0 value after 48 hours were determined to be 39 mg/l, the EC100 value are >203 mg/l. The EC50 (48 hrs.) value was found to be 146 mg/l. According to Directive 93/21/EEC (adaptation to Directive 67/548/EEC) the ecotoxicological classification of FAT 20011/D is not toxic to Daphnia magna after 48 hours.

Description of key information

FAT 20011/D was tested for its short term toxicity to Daphnia magna in a 48-hours static test. The study was performed under Good Laboratory Practice (GLP) conditions in Switzerland. The experiment was done according to the OECD Guideline for Testing of Chemicals, Section 2, No. 202, adopted 4th April 1984, (issued 1993) and EU-Guideline 92/69, EEC Nr. L 383 A, C. 2, (29th Dec. 1992).

The nominal concentrations tested were 45,100 and 220 mg test article/l, a vehicle and a blank control without any additions. The test concentrations were prepared from a 1 g/l stock solution. In all test concentrations non-alteration of the test substance was observed after 48 hours. During the test the test article concentrations in the analyzed test media were in the range of 87 - 95 % of the nominal values. Therefore, the reported results are related to the actual mean values of the analyzed concentrations of the test article in the test medium, measured at the beginning and at the end of the test. No immobilization and abnormal behavior of the Daphnia was observed in the controls. The NOEC and EC0 value after 48 hours were determined to be 39 mg/l, the EC100 value are > 203 mg/l. The EC50 (48 hrs) value was found to be 146 mg/l. According to Directive 93/21/EEC (adaptation to Directive 67/548/EEC) the ecotoxicological classification of FAT 20011/D is not toxic to Daphnia magna after 48 hours.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

146 mg/L