Octasodium 2,2'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino(1-hydroxy-3,6-disulphonatonaphthalene-2,8-diyl)azo]]bisnaphthalene-1,5-disulphonate

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

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)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

STORAGE:
Routinely, the samples were analysed immediately. Only in exceptional cases, they were stored overnight deep frozen and protected from light.

ANALYSIS OF SAMPLES:
Aliquots of the samples from the biological test were directly analysed by HPLC and UV/VIS-detection.

Vehicle:

no

Details on test solutions:

CULTURE AND DILUTION WATER
Reconstituted water (so-called 'M4 medium' according to OECD 202 and EC Method C.2, annex 1) was used for the maintenance of the test animals and the preparation of stock and test solutions of the test item.

PRE_TREATMENT OF TEST ITEM AND PREPARATION OF TEST ITEM CONCENTRATION
A stock solution was prepared to give the desired series of test concentrations. 129.9 mg of the test item (= 100 mg/L active ingredient) were added to 1 litre of dilution water and stirred for 1 h on a magnetic stirrer. The pH was measured to be 7.9.
To produce the different test item concentrations appropriate amounts of the stock solution were diluted with dilution water to a volume of 500 mL. 50 mL of the prepared solutions were taken per replicate and 0.5 mL of dilution water containing 5 daphnids was given to all replicates resulting in the final nominal concentrations. For each test item concentration and the control 4 replicates were prepared.

STANDARD SOLUTIONS USED FOR CALIBRATION OF THE TEST ITEM (HPLC ANALYTIC):
42.0 mg of the test item were dissolved and made up with Millipore water to the mark in a 100 mL volumetric flask to prepare a stock solution of 325.50 mg/L calculated as active ingredient. Defined volumes of this stock solution were diluted with Millipore water to obtain standard solutions in the range of 3.255 to 65.10 mg/L. The concentration of the lowest used calibration solution was employed as the limit of quantification. For the control, the fivefold amount was injected. Therefore, the limit of quantification can be specified as 0.651 mg/L. These solutions were used to calibrate the HPLC-system.

STANDARD SOLUTIONS USED FOR VERIFICATION OF THE CALIBRATION OF THE TEST ITEM AND BLANK TEST (HPLC ANALYTIC):
31.0 mg of the test item were dissolved and made up with Millipore water to the mark in a 100 mL volumetric flask to prepare a stock solution of 240.25 mg/L calculated as active ingredient. This solution was diluted with Millipore water to obtain a concentration of 12.01 mg/L. The daily verification of the calibration was performed with the diluted Standard Solution of Verification. The mean recovery rate was calculated.
Millipore water without the test item was analysed (analytical blank).

Test organisms (species):

Daphnia magna

Details on test organisms:

Name: Daphnia magna STRAUS, parthenogenetic females
Source: Strain of Bundesgesundheitsamt Berlin
Maintenance and Acclimatisation: A population of parthenogenetic females of synchronized age structure has been maintained for more than 20 years in the test facility under constant temperature conditions (20 +/- 2 °C) at a 16 : 8 hour light-dark photoperiod (light intensity: < 20 μE x m-2 x s-1). The culture water (so-called 'M4 medium') was partly renewed once a week. The Daphnia were exclusively fed unicellular green algae (Desmodesmus subspicatus) 'ad libitum'. Mortalities of parent Daphnia during the culture period were recorded daily. The neonates were separated from their parent Daphnia by filtration prior to the acute test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 d

Test temperature:

19.6-20.7 °C

pH:

7.9-8.0

Dissolved oxygen:

99-102 %
8.9-9.2 mg/L

Nominal and measured concentrations:

Nominal: 0.88, 1.9, 4.3, 9.4, 20.7, 45.5, 100 mg/L
Measured at 24 h: 1.003, 10.675, 52.651 mg/L
Measured at 48 h: 0.927, 10.413, 51.798 mg/L

Details on test conditions:

EXPOSURE CONDITIONS
-Test vessels: 100 mL glass beakers covered with watch glasses holding 5 neonates in 50 mL of test medium
-Experimental design:
-7 test concentrations plus 1 control,
-5 neonates per vessel,
-4 replicates per concentration/control,
-no feeding during the exposure period
-static system
-Method of initiation: neonates were placed in prepared media
-Photoperiod : 16 h light : 8 h dark
-Temperature of incubation unit: 19.2 to 20.0 °C
-Aeration: none
-Test item concentration/s: 0.88, 1.9, 4.3, 9.4, 20.7, 45.5 and 100 mg/L (active ingredient)
-Method of administration: stock solution
-Medium renewal: none
-Duration of exposure: 48 hours
-Criteria of effects: The criterion of adverse effects used in this study was the item-induced alteration of the normal mobility behaviour and the loss of locomotory actions of the neonates, observed at 24 and 48 hours.

RANGE FINDING TEST
A range finding test preceded the main test. It provided information about the range of concentrations which were used in the main test. The following nominal concentrations of the test item were tested in the range finding test: 0.1, 1, 10 and 100 mg/L.

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

34.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

behaviour

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

behaviour

Details on results:

The results are expressed in terms of nominal concentrations (active ingredient).
Effective concentrations ranged from 113.6 % to 115.7 % of nominal values at 0 hours and from 105.3 % to 113.8 % of nominal values at 48 hours.

Validity criteria fulfilled:

yes

Remarks:

immobilisation and other abnormalities in the controls did not exceed 10% by the end of the test; dissolved oxygen concentration remained above 3 mg/L throughout the exposure period

Conclusions:

Acute toxicity to daphnia (daphnia magna) of Reactive Red 141 was tested in a static test. After after 48 hours an EC50 of 34.7 mg/L was determined. The results are expressed in terms of nominal concentrations.

Executive summary:

A study was performed to assess the acute toxicity of Reactive Red 141 to Daphnia magna STRAUS under static conditions.

The study was conducted in accordance with the Council Regulation (EC) No 440/2008, Method C.2 ‘Acute toxicity for Daphnia’ (2008) which is equivalent to OECD Guideline for Testing of Chemicals No. 202 'Daphnia sp., Acute Immobilisation Test' (adopted April 13, 2004).

The Daphnia were exposed to a range of concentrations, nominally 0.88, 1.9, 4.3, 9.4, 20.7, 45.5 and 100 mg/L of the active ingredient of Reactive Red 141 dissolved in dilution water. Auxiliary used to prepare the test media was a magnetic stirrer.

Observations were made on the swimming ability and the immobilisation rate, respectively, after 24 and 48 hours of exposure. A 48hr-EC50 value of 34.7 mg/L was determined.

The results are expressed in terms of nominal concentrations (active ingredient). Effective concentrations ranged from 113.6 % to 115.7 % of nominal values at 0 hours and from 105.3 % to 113.8 % of nominal values at 48 hours.

This toxicity study is classified as acceptable and satisfies the guideline requirements for the acute daphnia study.

Description of key information

Acute toxicity to daphnia (daphnia magna) of Reactive Red 141 was tested in a static test according to OECD Guideline 202. After 48 hours an EC50 of 34.7 mg/L was determined. The results are expressed in terms of nominal concentrations.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

34.7 mg/L