Disodium 3,3'-[[6-[bis(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diyl]bis[imino(3-methoxy-4,1-phenylene)azo]]bis[benzenesulphonate]

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

key study

Study period:

From March 03 to April 15, 1989

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 has been detailed in the report attached to the IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

adopted April 4, 1984

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Analytical Chemistry: stability of the test substance under test conditions as determined by analysis of water samples taken from the test media during the range finding test. During the main study actual concentration of the test substance were determined by analysis of duplicate samples taken at t 0 hour.
- Sampling regime frequency during the range finding: three test concentrations: 0.1, 10 and 1000 mg/l in the range finding study at t 0, 24 and 96 hours.
- Sampling regime frequency during the main study: in duplicate from the test concentration of 1000 mg/l at t 0 hour.
- Sampling volume: 50 ml from the approximate centre of the test vessels.

Details on test solutions:

For formulation of the test concentrations used in the main study a stock soiution of 10 g test substance in 2 Iitres was used.
The stock solution was treated with ultra sonication for 10 minutes to accelerate the dissolving of the test substance.
Subsequentiy it was added to 8 liters of tap water.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: carp.
- Source: young Cyprinus carpio were supplied by Zodiac, Wageningen, The Netherlands.
- Length at study initiation: between 2 - 4 cm, deviation of 0.5 cm.

ACCLIMATION
- Acclimation period: the fish were adapted to the test medium without test substance for at least seven days following a 48 hour settling in period.
- Test vessel: carp held in a 100 l glass aquaria (bonded with silicone rubber).
- Medium: aquaria were continuously filtered (4 l/min) over synthetic wool fiber (Perion) and active carbon with washed fine gravel covering the bottom of the aquaria. Replacement of filters and water: once a month if necessary.
- Photoperiod: for 14 hours a day.
- Temperature: 23 ± 2 °C
- Health during acclimation: in the batch of which fish were used for the test, mortality during seven days prior to the start of the test was less than 5 %.

FEEDING DURING TEST
- Food type: Artemia or Trouvit OD.
- Frequency: once a day.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

20 ± 2 °C; during the main study room temperature dropped below 20 °C due to a technical failure. Hence, the temperature of the test medium varied between 18.5 and 19.5 °C. Although the temperature of the media was not optimal, no adverse effects were recorded in the blank control.

pH:

The pH ranged for all concentrations during the whole study from 7.9 to 8.6.

Dissolved oxygen:

Oxygen concentration in the test media was found to be higher than 5 mg/l for all measurements performed during the main study.

Details on test conditions:

TEST SYSTEM
- Test vessel: 10 litres, all-glass.
- Aeration: continuously aerated.
- No. of organisms per vessel: 10 fish per vessel.
- No. of vessels per concentration: 1 replicate.
- No. of vessels per control: 1 replicate.

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: tap water.

OBSERVATIONS
- Mortality: 5, 24, 48, 72 and 96 hours after start of exposure.
- Dissolved oxygen content and pH: at time 0, 24, 48, 72 and 96 hours starting from the introduction of the fish.
- Water temperature: in the blank control vessels at time 0, 24, 48, 72, 96 hours using a thermometer.

Reference substance (positive control):

yes

Remarks:

pentachlorophenol

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

After 96 hours of exposure no effects or mortality were observed.

ACTUAL VERSUS NOMINAL CONCENTRATIONS
Analysis of samples taken during the range finding study showed that the test substance was stable in water for minimal 96 hours. In samples taken at t 0 from the nominal concentration of 1000 mg/l the actual concentration was 90 %. At t 24 hours and 96 hours the actual concentrations measured were respectively 98 and 103 % of the nominal concentration.
Analysis carried out with the samples taken during the main study at t 0 hours from the nominal concentration of 1000 mg/l showed that the actual concentration was 91 %.

Sublethal observations / clinical signs:

Oxygen content and pH conditions

Vessel	Sampling time
	0 hr	24 hrs	24 hrs	72 hrs	96 hrs
pH
Control	7.9	8.5	8.3	8.6	8.4
1000 mg/l	8.6	8.5	8.3	8.6	8.3
Oxygen concentration (mg/l)
Control	7.6	6.8	8.8	9.4	8.8
1000 mg/l	7.6	7.0	8.3	9.4	8.6

Validity criteria fulfilled:

yes

Remarks:

Constant conditions were maintained throughout the test; mortality in the control did not exceed 10 % at the end of the test and dissolved oxygen has been at least 60 % of air saturation value

Conclusions:

LC50 (96h) > 1000 mg/l (nominal)

Executive summary:

The short-term toxicity to fish potential of the test substance was tested following the method and procedures outlined into the OECD guideline 203.

Based on the range-finding test results, in the main study ten fish per concentration were exposed for 96 hours in a static system to the limit concentration of 1000 mg/l. Stability of the test substance under test conditions was examined by analysis of samples taken during the range finding study; at the nominal concentration of 1000 mg/l, the actual concentration was 90 % to 103 % of the nominal concentration throughout the exposure period of 96 hours.

In the main experiment, after 96 hours of exposure, no effects or mortality were observed. Analysis of duplicate samples taken at the beginning of the main study showed that the actual concentration was 91 % of the nominal concentration.

Conclusion

The 96h-LC50 appears to be higher than 1000 mg/l (nominal).

Description of key information

Non harmful/toxic to fish for short-term exposure (LC50 (96h) > 1000 mg/l nominal).

Additional information

There is no information about the short-term toxicity to fish of Direct Yellow 142, thus the available data on structural analogous Similar Substance 01 has been taken into account. The read across approach can be considered as reliable and adequate for the purpose; details and explanations are detailed in the report attached to the IUCLID section 13.

The short-term toxicity to fish potential of the Similar Substance 01 was tested following the method and procedures outlined into the OECD guideline 203.

In the main study ten fish per concentration were exposed for 96 hours in a static system to the limit concentration of 1000 mg/l. After 96 hours of exposure no effects or mortality were observed. Analysis of duplicate samples taken at the beginning of the main study showed that the actual concentration was 91 % of the nominal concentration.