Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-[(2-cyanoethyl)[2-(2-hydroxyethoxy)ethyl]amino]-1,3,5-triazine-4,2-diyl]imino]]bis[benzene-1,4-disulphonate]

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 August the 28th to September the 08th, 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:

Justification for read across is detailed in 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 on 4 April, 1984

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Sampling frequency: in duplicate from the approximate centre of the vessel at t=0, t=2 and just before the first renewal (t=24).
- Sampling volume: 5 ml from the approximate centre of the test vessels.
- Storage: the samples were directly analysed.
- Stability: stability of the test substance under test conditions was determined also during the pilot test. Duplicate 5 ml samples were taken at t = 0, 24 and 96 hours.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp.
- Source: Zodiac, proeffacc, "De Haar Vissen", L.U.
- Length at study initiation: 2.78 ± 0.12 cm
- Weight at study initiation: 0.45 ± 0.05 g.
- Feed: Trouvite or Artemia.
- Frequency: once a day.

ACCLIMATION
- Acclimation period: after delivery, the fish will be held in 100-liter tanks for at leas 14 days.
- Adaption: prior to the testing, the fish will be adapted to the test medium without test substance for at leas 7 days following a 48 hours settling period.
- Medium: filtered and areated tap water is supplied continuosly. Evaluation of contaminant analysis was performed.
- Temperature: 23 ± 2 °C

VALIDITY OF BATCH
In the batch of which fish were used for the test, mortality during seven days prior to the start of the test has to be less than 5%. If the incidence of mortality is between 5 and 10 % acclimatization is continued for seven additional days. If mortality is higher than 10%, possible rejection of the entire batch will have to be considered.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

Ranged from 21 to 22 °C.

pH:

Ranged from 8.0 to 8.6.

Dissolved oxygen:

Ranged from 8.4 to 9.6 mg/l

Nominal and measured concentrations:

1000 mg/l nominal

Details on test conditions:

TEST SYSTEM
- Test vessel: 10 litres; all-glass.
- Renewal rate of test solution: every 24 h.
- No. of organisms per vessel: 10
- No. of vessels per concentration: 3 replicates.
- No. of vessels per control: 1 replicate.
- Biomass loading rate: 0.75 g fish per litre test medium.

OTHER TEST CONDITIONS
- Photoperiod: 14 hours light daily
- Feeding: no from 24 hours prior to the test and during the total test period.

RECORDINGS
- Mortality: checked at 5.5, 24, 48, 72 and 96 hours after treatment.
- Fish size and weight: 10 fish of the batch of fish, from which fish were used in the study, were weighted and measured prior to the start of the test.
- D.O. and pH: checked at 0, 24, 48, 72 and 96 hours startin from the introduction of the fish.
- Water temperature: checked at 0, 24, 48, 72 and 96 hours in the blank control vessel.

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:

No mortality or other effects were observed at the 1000 mg/l concentrations and the blank.

MEASURED CONCENTRATIONS
Analysis of the samples taken during the main test in the period before the first renewal revealed actual concentrations ± 10 % relative to the nominal concentrations of 1000 mg/l.

Sublethal observations / clinical signs:

Incidence of mortality observed in the main study

Nominal concentration [mg/l]	Number exposed	Number of dead fish recorded after
		5.5 h	24 h	48 h	72 h	96 h
Blank	10	0	0	0	0	0
1000 A	10	0	0	0	0	0
1000 B	10	0	0	0	0	0
1000 C	10	0	0	0	0	0

pH levels after various time intervals during the main study

Nominal concentration [mg/l]	pH values
	0 h	24 h		48 h		72 h		96 h
	fresh	old	fresh	old	fresh	old	fresh	old
Blank	8.6	8.1	8.0	8.4	8.4	8.4	8.4	8.5
1000 A	8.6	8.2	8.1	8.4	8.4	8.5	8.4	8.5
1000 B	8.6	8.2	8.1	8.4	8.4	8.4	8.5	8.5
1000 C	8.6	8.1	8.1	8.4	8.4	8.4	8.5	8.5

Oxygen concentrations after various time intervals during the main study

Nominal concentration [mg/l]	oxygen concentration [mg/l]
	0 h	24 h		48 h		72 h		96 h
	fresh	old	fresh	old	fresh	old	fresh	old
Blank	8.6	8.8	9.6	9.2	9.4	9.2	9.3	8.4
1000 A	8.7	8.8	9.3	9.1	9.4	9.2	9.3	8.4
1000 B	8.9	8.8	9.2	9.0	9.3	9.2	9.4	8.5
1000 C	8.8	8.8	9.4	9.1	9.4	9.0	9.4	8.6

Concentration of test substance in the test medium (stability assessment)

Time of analysis	Concentration [mg/l]
	prepared	replicate	analyzed	%
t = 0 h	1000	A	969	97
		B	983	98
		C	1010	101
t = 2 h	1000	A	940	94
		B	952	95
		C	972	97
t = 24 h	1000	A	919	92
		B	949	95
		C	962	96

Validity criteria fulfilled:

yes

Remarks:

mortality of the control < 10 % at the end of the test; conditions satisfactorily manteined troughout the test; test substance concentration satisfactorily mantained (± 20 % of nominal concentration)

Conclusions:

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

Executive summary:

A fish limited acute toxicity test was performed according to OECD 203. Test duration was 96 hours and consisted of a semi-static system with renewal time of 24 hours. Analysis of the samples taken during the test in the period before the first renewal revealed actual concentrations ± 10 % relative to the nominal concentrations of 1000 mg/l.

The test is considered valid since it met all the validity criteria of the followed guideline.

Under the test conditions, no adverse effect was recorded on the test organisms and therefore the test substance can be considered to be not acutely toxic against fish, i.e. NOEC (96 h) higher/equal to 1000 mg/l.

Conclusion

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

Description of key information

Not harmful/toxic for short-term exposure to fish

Additional information

The substance was tested for short-term toxicity potential to fish (rainbow trout) following internal method and procedures. 500 mg/l was indicated to be the maximum concentration at which all experimental animals survive, after an exposure period of 48 hours. Unfortunately, a complete test report is not available and only a sheet of test results can currently be retrieved; in addition, the lot tested was characterized by a limited content of Optical Brightener 380. Therefore, a reliability cannot be assigned and the test has been included just for completeness sake.

In order to investigate the short-term toxicity potential to fish of Optical Brightener 380, the available experiment conducted on the structural analogous Similar Substance 01 has been taken into consideration. The read across approach can be considered reliable and appropriate to investigate the property (details for the approach are included into the IUCLID section 13).

A fish acute toxicity limit-test was performed on Similar Substance 01, according to OECD 203. Test duration was 96 hours and consisted of a semi-static system with a renewal time of 24 hours. Analysis of the samples taken during the test in the period before the first renewal revealed actual concentrations ± 10 % relative to the nominal concentrations of 1000 mg/l.

Under the test conditions, no adverse effect was recorded on the test organisms, in none of the replicates assayed.