Tetrasodium 7,7'-(carbonyldiimino)bis[4-hydroxy-3-[(2-methyl-4-sulphonatophenyl)azo]naphthalene-2-sulphonate]

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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Study period:

March from 13th to 15th, 2007

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Justification for type of information:

The read across approach is detailed into the document attached to the IUCLID section 13.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

adopted 13 April 2004

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

no

Analytical monitoring:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Test organism: Daphnia magna (Straus, 1820).
- Size: < 2mm

BREEDING SYSTEM
- Breeding: parental and young Daphnia were held in 250 I glass aquaria
- lllumination: 16 h per day
- Temperature: 20 ± 2 °C
- Feed: 15 g sera micropan (sera GmbH, D-52525 Heinsberg) homogenized in 1 L deionised water
- Frequency of feeding: once a day, except weekends
- Medium: continuously aerated reconstituted water according to ISO (3); prepared with distilled water (conductivity < 1.5 µS/cm).
The final test medium contained 25 ml/l of each of the following four stock solutions:
CaCl2 × 2 HzO 1 1.76 g/l
MgSO4 × 7 H2O 4.93 g/l
NaHCO3 2.59 g/l
KCl 0.23 g/l
The sum of the Ca and Mg ions in this solution is 2.5 mmol/l
The ratio of Ca:Mg ions is 4:1 and of Na:K ions is 10:1
The total alkalinity of this solution is 0.8 mmol/l
The bottom of the aquaria is covered with gravel (≤ 1.5 cm). Every month ,25 % of the total volume of the aquaria is replaced by fresh water.

ACCLIMATION
- Acclimation period: daphnia were acclimatized to the reconstituted water for 4 hours before introduction into the test media.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 ± 0.5 °C (temperature-controlled water bath).

Dissolved oxygen:

Range: 5.8 - 6.2 at 24 h; 6.0 - 6.3 at 48 h

Nominal and measured concentrations:

Nominal: 1, 10, 100 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 ml flasks, all-glass, with 50 ml of test medium.
- Aeration: not aerated.
- No. of organisms per vessel: 20 per vessel.
- No. of organisms per concentration: 40 individuals per test concentration.
- No. of vessels per concentration: 2 replicates.
- No. of vessels per control: 3 replicates.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water (lSO 6341).
- Aeration: prior to the test, a batch of reconstituted water was aerated vigorously over a period of 2 h. Final test concentrations were prepared with this aerated water by dilution of stock solutions of the test substance.

OTHER TEST CONDITIONS
- Adjustment of pH: the pH was not adjusted before the test.
- Light: 16 h photoperiod a day, supplied by overhead white fluorescent tubes.

OBSERVATION AND MEASUREMENTS
- Temperature: determinated in a control flask at the start and at the end of the test.
- pH: determined in all vessels prior to the addition of Daphnia to the test medium and at the end of the test.
- Dissolved carbon: determined in all vessels prior to the addition of Daphnia to the test medium and at the end of the test.
- Observation: observation of immobile Daphnia were made after 24 and 48 h of exposure.

VALIDITY OF THE TEST
The test is considered valid if not more than 10 % Daphnia in the control have been immobilized or trapped al the surface of the water. The dissolved oxygen concentration at the end of the test should not be < 60 % of lhe air saturation value at the temperature used.

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

At all nominal test concentrations no significant effects (<10 % immobilization) were observed after 24 and 48 h of exposure. No significant effects were also observed in the control after 24 and 48 h of exposure. The no-observed-effect concentration (EC0) for 24 and 48 h of incubation resulted to be higher than 100 mg/l.
100 % immobilization was not detected.

Validity criteria fulfilled:

yes

Conclusions:

EC50 (48h) > 100 mg/l (nominal)

Executive summary:

The acute toxicity of test item to Daphnia magna was investigated under static exposure conditions over a period of 48 h. A screening test with nominal concentrations of 100, 10 and 1 mg/l, respectively, was performed. 40 organisms, divided in 2 test vessels, were exposed to each concentration of test substance. The final test concentration was prepared by dilution of a stock solution of test substance. No analytical verification of test material was conducled. The evaluation of the test was based on the nominal concentrations of the test material, assuming the test compound to be stable in water over 48 h.

At all nominal test concentrations, no significant effects (<10 % immobilization) were observed after 24 and 48 h of exposure. No significant effects were also observed in the control after 24 and 48 h of exposure.

The No-Observed-Effect Concentration (NOEC) for 24 and 48 h of incubation resulted to be higher than 100 mg/l. 100 % immobilization was not detected.

Conclusion

EC50 (48h) > 100 mg/l (nominal)

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for type of information:

The read across approach is detailed into the document attached to the IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

At exposure initiation, eight samples of the test item concentrations and the control were collected. One of them was transferred for chemical analyses at exposure initiation, whereas the others were located under test conditions and transferred for chemical analyses at exposure termination.

Details on test solutions:

PREPARATION OF TEST SOLUTIONS:
The test item in the amount of 59.9 mg was weighed in a glass beaker and quantitatively transferred into flask by multiple washing the Elendt M7 medium and filled up to 500 ml. The test item concentration of 120 mg/l was visually heterogeneous. Therefore, the content of flask was sonicated for 5 minutes. Then, the test item concentration of 120 mg/l was visually homogeneous. From the test item concentration and the control, 50 ml were collected and transferred for chemical determinations.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Test organism: Daphnia Magna Straus.
- Source: daphnids were originated from the standard laboratory culture cultivated at the Institute of Industrial Organic Chemistry, Branch Pszczyna, Department of Ecotoxicology, Laboratory of Aquatic Toxiology.
- Age of organisms: only organisms up to 24 hours old (not first brood progeny) and in good physiological condition were used for exposure.
- Feeding during test: Daphnia magna were not fed.

CULTURING
- Vessels: Daphnia magna was cultured in glass beakers with a capacity of 150 ml (one parent per vessel).
- Medium: 100 ml of Elend M7 medium.
- Temperature: room temperature ranging from 19 to 22 °C.
- Photoperiod: daily cycle 16 h light : 8 h dark.
- Food type: Daphnia magna were fed with a suspension of algae; meixture of two species Pseudokirchneriella subcapitata: Desmodesmus subspicatus (in 2:1 ratio) originating from separate cultures in the Laboratory of Aquatic Toxicology. Group B vitamins and micronutrients necessary for proper growth were supplied with the lyphilixe suspension of Spirulina sp.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

20.2 - 20.9°C

pH:

Start of the test : 7.45
End of the test: 7.28

Dissolved oxygen:

Start of the test: 9.0-9.1 mg/l
End of the test: 8.8-8.9 mg/l

Nominal and measured concentrations:

Nominal concentration: 0 (control) and 120 mg/l (based on preliminary test)

Details on test conditions:

TEST SYSTEM
- Test vessel:glass beakers of 150 ml capacity.
- Type: closed; the beakers were covered with transparent lids in order to minimize evaporation and to prevent accidental contamination.
- No. of organisms per vessel: 5 organisms per replicate.
- No. of vessels per concentration: 8 replicates.
- No. of vessels per control: 8 replicates.

OTHER TEST CONDITIONS
- Photoperiod: each test was conducted with daily cycle 16 h light : 8 h dark.
- Light intensity: fluorescent lighting was used.

EFFECT PARAMETERS MEASURED:
Daphnia magna was observed for immobilization after 24 and 48 h of exposure.

MEASUREMENTS
During exposure, the temperature was continuously recorded using a sensor submerged in an additional test vessel. The pH values and the dissolved oxygen concentrations were measured at exposure initiation, i.e. before splitting up into replicates, and at exposure termination in pooled replicates.

VALIDITY CRITERIA
- The immobilization of Daphnia magna in the control should be not more than 10 %
- The dissolved oxygen concentrations in the test vessels should be not less than 3 mg/l

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No signs of immobilisation were observed throughout the test.

MEASURED CONCENTRATIONS
- Measured concentration at Exposure initiation: < LoD (control) and 106.00 mg/l
- Measured concentration at Exposure terminiation: < LoD (control) and 100.16 mg/l

Results with reference substance (positive control):

The EC50 (48h) of the reference substance potassium dichromate was 0.66 mg/l and within the range of 0.58 and 0.76 mg/l.

Reported statistics and error estimates:

Since the immobilization of Daphnia magna was less than 10 %, no statistical analysis was needed.

Validity criteria fulfilled:

yes

Remarks:

the immobilization in the control was 0 % and the dissolved oxygen concentrations in the test vessels were within the range of 8.8 -9.1 mg/l

Conclusions:

EC50 (48h) > 120 mg/l (nominal)

Executive summary:

The substance was assessed using Daphnia magna in a 48-hour Acute Immobilisation Test. Based on results obtained in a preliminary experiment, organisms were exposed to a single concentration of 120 mg/l (limit concentration), over a period of 48 hours. A concurrent control group was run. The analytically measured test item concentration remained within ± 20 % of the nominal during the test period of 48 hours; therefore, biological results are based on the nominal concentration.

There was no immobilisation observed in the daphnids exposed to either the test item or in the control group. The 48-h NOEC was determined to be equal/higher than 120 mg/l.

Conclusion

EC50 (48h) > 120 mg/l (nominal)

Description of key information

Not harmful/toxic to invertebrates for short-term exposure (EC50 (48h) > 120 mg/l, nominal).

Additional information

There are no information about the short-term toxicity potential to aquatic invertebrates of Direct Orange 118, thus the available data on structural analogues Similar Substance 01 and Similar Substance 02 was taken into account. The read across approach can be considered as reliable and suitable for the purpose; details are available in IUCLID section 13.

The Similar Substance 02 was assessed using Daphnia magna in a 48-hour Acute Immobilisation Test. Based on results obtained in a preliminary experiment, organisms were exposed to a single concentration of 120 mg/l. There was no immobilisation observed in the daphnids exposed to either the test item or in the control group, therefore, the 48-h NOEC was determined to be equal/higher than 120 mg/l. The analytically measured test item concentration remained within ± 20 % of the nominal during the test period of 48 hours; therefore, biological results are based on the nominal concentration (Institute of Industrial Organic Chemistry Branch Pszczyna, 2015).

In the supporting study, the acute toxicity of Similar Substance 01 was assayed under conditions comparable to those above described. A screening test with nominal concentrations of 100, 10 and 1 mg/l was performed. At all nominal test concentrations, no significant effects (less than 10 % immobilization) were observed after 24 and 48 h of exposure (Häner, 2002).