4,4'-[(6-chloro-1,3,5-triazine-2,4-diyl)diimino]bis[5-hydroxy-6-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonic] acid, sodium salt

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

10 December 1991 to 12 December 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

84/449/EEC

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

The concentration of V723 in the test solutions was measured at the start (excess solution) and end of the test. At the end of the test one replicate of each test concentration was sampled.

Details on test solutions:

A solution of 112 mg V723/litre was prepared by addition of V723 to the dilution medium.

Test organisms (species):

Daphnia magna

Details on test organisms:

The test organism was the freshwater crustacean Daphnia magna Straus, obtained from continuous laboratory cultures.
The stock cultures of Daphnia were maintained in a reconstituted water medium, identical to the test dilution water, at a temperature of 20 ± 2°C, and a photoperiod of 16 hours light: 8 hours dark.
The cultures were fed a defined diet of algae Chlorella vulgaris and yeast.
Culture conditions were such that the Daphnia reproduction was by diploid parthenogenesis.
Daphnia <24 hours old were used for testing.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No post exposure observation

Hardness:

Hardness (total as CaCO3) 179 mg/l

Test temperature:

The temperatures recorded at 0, 24 and 48 hours were 19.8, 19.5 and 19.8°C respectively

pH:

8.4-8.5

Dissolved oxygen:

9.0 - 9.5 mg/l

Salinity:

Not applicable.

Nominal and measured concentrations:

A solution of 112 mg V723/litre was prepared by addition of V723 to the dilution medium. Information supplied with the test sample V723 indicated that it contained 10.5% of water and test concentrations were adjusted accordingly to account for a nominal test concentration of 100 mg/l.

Details on test conditions:

Borosilicate glass beakers were used as test vessels, with four replicates per exposure concentration, each containing 200 ml of test solution.
When the test solutions were at test temperature, five Daphnia were added to each vessel in <2.0 ml of dilution water, giving a total of 20 Daphnia per concentration.
The temperature was maintained at 20 ± 1°C and a photoperiod of 16 hours light: 8 hours dark was provided.
The Daphnia were not fed during the course of the test.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The 24 and 48 hour median effective concentrations of the test substance to the freshwater crustacean Daphnia magna (<24 hours old) were determined under static test conditions at a temperature of 20 ± 1°C.
The median effective concentration (EC50) is defined as the concentration, calculated from the data obtained, resulting in immobilisation of 50% of the Daphnia in the time period specified.
The values obtained were:

EC50 (mg/l)
24 hour >100
48 hour >100

The values obtained were based on nominal concentrations
The overall mean of the measured concentrations of the test material as determined at the start and finish of the test was 90% of the nominal value.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

Not reported.

Daphnia response

Time	Nominal	Number affected per				Total	Total	%
(hours)	conc		replicate			number	number	affected
	(mg/1)	A	B	C	D	tested	affected
24	Control	0	0	0	0	20	0	0
	112	0	0	0	0	20	0	0
48	Control	0	0	0	0	20	0	0
	112	0	0	0	0	20	0	0

Validity criteria fulfilled:

yes

Conclusions:

As no effect was found, the 24 and 48 hr EC50 values were >112 mg/L

Executive summary:

Study conducted to EU test guidance 84/449/EEC part C2 in compliance with GLP.

No effect was found at a concentration of 112 mg/L after 48 hours. The EC50 is > 112 mg/L. The substance is not considered to pose a hazard to Daphnia.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 October 1989 to 18 January 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study conducted to EU test guidance in compliance with GLP.

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Test concentrations: 0 / 560 / 1000 mg/L

Vehicle:

no

Details on test solutions:

The relevant amount of the test substance was weighed into a graduated flask (200 mL), filled up with test water and stirred for about 5 minutes with a magnetic stirrer. The test batches were present as red solutions. No segregation or formation of precipitates could be observed.
In order to determine the test substance concentration, water samples were taken at study end.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna (STRAUS)
Origin: Stock culture of Department of Toxicology (Hoechst AG)
Breeding conditions: beakers (height 235 mm, diameter 128 mm), filled with 2L test water (according ISO/DIS 7346/1) served as culture vessels. 20 adult daphnids were used per culture vessel. Animals were transferred to fresh prepared vessels once a week. The room temperature was at 20 ± 1 °C. The breeding room was illuminated by diffuse light in a day/night cycle of 12 hours. The animals were fed with monocellular green algae (Scenedemus subspicatus) and a fine suspension of Tetra min dry food (Tetra Werke, Melle).

To collect test animals with an age of at least 6 hours and a maximum of 24 hours, approx. 3 weeks old parent animals were separated in a cristallising dish (3L volume) filled with test medium for 18 hours. After 18 hours the adults were separted from the young daphnids by sieving through sieves of different mesh size. Adults remain on the upper crude sieve while the young daphnids are found on the fine sieve. These animals were used for the study after further 6 hours.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No post exposure observation period

Hardness:

No data

Test temperature:

20.4 - 21.3 °C

pH:

7.7 - 8.3

Dissolved oxygen:

8.6 - 9.5 mg/L

Salinity:

Not applicable.

Nominal and measured concentrations:

0, 560 and 1000 mg/L.

Details on test conditions:

The study was conducted in 2 stages. 20 daphnids, divided into two parallel groups of 10 animals, were used for each concentration.

Water for dilution
Reconstituted water, composition according to ISO/DIS 7346/1 was used as water for dilution.
Preparation was carried out in a unit consisting of two Hostalen®- lined steel vessels with a capacity of 1700 litres each.

The dilution water was prepared as described and was aerated until oxygen saturation. The pH was measured before the use of each study batch. It was at 8.3.

Test conditions
The study was conducted in a static system. As test chambers 300 mL cristallising dishes were used (height 55 mm, diameter 95 mm, water level about 32 mm), each filled with 200 mL test water. The room temperature was about 21 °C. The room was illuminated by fluorescent tubes for 12 hours each day.

Test procedure
After the test concentrations had been prepared and water parameters were recorded, 10 daphnids were used in each test chamber. The daphnids were not fed during the entire study period. Inspection of the daphnids took place after 24 and 48 hours and involved recording the immobility and further possible deviations to the control group. Immobile daphnids were removed from the chambers. Those animals which were not able to swim within 15 seconds after gentle agitation of the test container were considered to be immobile. The water parameters were measured after adding the substance and at the end of the test.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Probit analysis could not be carried out due to the lethality rate. As the factor for the concentration steps was 1.8 no further concentrations were tested. The EC50 range was given.

Under the conditions of this test with Reaktiv-Rot F-66 813 FW the following immobility rates were observed after 24 and 48 hours:

Concentration (mg/L) 24 hours 48 hours
Absolute % Absolute %
0 0/40 0 0/40 0
560 0/20 0 0/20 0
1000 0/20 0 2/20 10
EC0 (mg/l)
highest tested conc. With no immobility 1000 560
EC50 (mg/L) > 1000 > 1000

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

No data

Immobility – individual values

Concentration (mg/L)	Test-chamber No.	Study-start (date)	Immobility hours after study start
			24	48
0	80a	10-Jan-90	0/10	0/10
0	80b	10-Jan-90	0/10	0/10
1000	36a	10-Jan-90	0/10	0/10
1000	36b	10-Jan-90	0/10	2/10
0	90a	16-Jan-90	0/10	0/10
0	90b	16-Jan-90	0/10	0/10
560	1a	16-Jan-90	0/10	0/10
560	1b	16-Jan-90	0/10	0/10

 

Validity criteria fulfilled:

yes

Conclusions:

EC50 > 1000 mg/L

Executive summary:

The study on acute toxicity to daphnids with the structural analogue (SA01) meets the requirements of EEC Directive 79/831 Annex V Methods for the determination of ecotoxicity 5.1.2 Acute toxicity for daphnia and the OECD Guidelines for Testing of Chemicals. The study was conducted in compliance with GLP.

Testing of SA01 for acute toxicity in Daphnia magna (water flea) over a total of 48 hours yielded the following immobility values:

 

after 24 hours (mg/L)    after 48 hours (mg/L)

EC_o      1000                                     560

EC₅₀       > 1000                                 > 1000

 

The concentrations tested were 0 / 560 / 1000 mg/L. Immobility was observed at 1000 mg/L.

The test batches were present as red solutions. No segregation or formation of precipitates could be observed.

 

In order to determine the substance concentration, water samples were taken at study end. All mean values were above 80 % of the nominal concentration.

Description of key information

EC50 > 1000 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

> 1 000 mg/L

Additional information

The study on acute toxicity to daphnids with the structural analogue (SA01) meets the requirements of EEC Directive 79/831 Annex V Methods for the determination of ecotoxicity 5.1.2 Acute toxicity for daphnia and the OECD Guidelines for Testing of Chemicals, Section 2 - Effects on biotic systems 202 Daphnia sp., 14-day reproduction test (including an acute immobilisation test). The study was conducted in compliance with GLP.

The concentrations tested were 0 / 560 / 1000 mg/L. Immobility was observed at 1000 mg/L.

The test batches were present as red solutions. No segregation or formation of precipitates could be observed.

In order to determine the substance concentration, water samples were taken at study end. All mean values were above 80 % of the nominal concentration.

Testing of SA01 for acute toxicity in Daphnia magna (water flea) over a total of 48 hours yielded the following immobility values:

 

	after 24 hours (mg/L)	after 48 hours (mg/L)
EC0	1000	560
EC50	> 1000	> 1000