Reaction products of 2,4,6-trichloro-1,3,5-triazine copuled with 3-aminopropyldiethylamine, coupled with 3-acetamidoanilinium chloride, diazotised, coupled with 2,5-Xylidine, coupled with 3-amino-4-hydroxybenzenesulphonamide, further coupled with resorcinol and metallized with iron chloride.

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From July 18th to August 26th, 1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

September 1982

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on test solutions:

The test solutions were prepared by adding appropriate amounts of a stock solution of the test material to test water, keeping the volume of the test solution at 4 litres in each test vessel.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish.
- Source: Stubenvoll, Allschwil, Switzerland.
- Length at study initiation: 2.75 cm
- Weight at study initiation: 0.62 g

ACCLIMATION
- Acclimation period: for a minimum of 12 days
- Acclimation conditions: 100 litres glass tanks in aerated test water of 23 ± 2 °C, under flow-through conditions.
- Feeding frequency during acclimation: fed daily util 24 hours before starting the test.

Test type:

not specified

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

Test vessels: 23 - 25 °C
Control: 23 - 25 °C

pH:

Test vessels: 7.9 - 8.2
Control: 8.0 - 8.1

Dissolved oxygen:

Test vessels: 66 - 92 %; 5.4 - 7.7 mg/l
Control: 77 - 95 %; 6.4 - 7.8 mg/l

Nominal and measured concentrations:

Nominal 1, 1.8, 3.2, 5.6 and 10 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: glass tanks (150 × 200 × 200 mm) with 4 l test solution.
- Aeration: none.
- No. of organisms per concentration: 12 fish.
- No. of vessels per concentration: 2 replicates.
- No. of vessels per control: 2 replicates.
- Biomass loading rate: 6 fish per 4 l test solution, i.e. 0.93 g fish/l.

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: carbon filtered drinking water with a total hardness of about 180 mg CaCO3/l and pH 7.6 to 8.0.

OTHER TEST CONDITIONS
- Photoperiod: fluorescent lighting.

PARAMETERS MEASURED
Dissolved oxygen, pH and temperature were measured daily in each test vessel.

EFFECT PARAMETERS MEASURED
The cumulative percentage mortality was plotted against the concentration on logarithmic-probability paper. A line was fitted, by eye, and the concentration corresponding to the 50 per cent response read off.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.5 mg/L

Nominal / measured:

estimated

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The maximum concentration producing no mortality is 1.65 mg/l
The minimum concentration producing 100 % mortality is 3.5 mg/l

Sublethal observations / clinical signs:

Rate of deaths

Test concentration (mg/l)	No of fish tested	No of deaths after				Mortality in % after 96 hrs
		24 hrs	48 hrs	72 hrs	96 hrs
Control	12	0	0	0	0	0
1	12	0	0	0	0	0
1.8	12	0	0	0	0	0
3.2	12	0	0	0	0	0
5.6	12	11	12	12	12	100
10	12	12	12	12	12	100

TEST CONDITIONS

Maintenance of test material

Nominal concentration (mg/l)	At beginning of test		At end of test		Geometrical mean
	mg/l measured via exstinction	% of nominal	mg/l measured via exstinction	% of nominal	mg/l measured via exstinction	% of nominal
Control	-	-	-	-	-	-
1	0.85	85.0	0.59	59.0	0.7	70.8
1.8	1.6	88.9	0.86	47.8	1.2	65.2
3.2	2.7	84.4	1.65	51.6	2.1	66.0
5.6	4.75	84.8	3.51	62.7	4.1	72.9
10	8.35	83.5	8.10	81.0	8.2	82.2

Test concentration (mg/l)	O2 (mg/l)					% oxygen saturation
	0 hrs	24 hrs	48 hrs	72 hrs	96 hrs	0 hrs	24 hrs	48 hrs	72 hrs	96 hrs
Control	7.8	6.6	6.4	6.5	6.8	95.0	78.0	77.0	79.0	80.0
1	7.7	5.9	6.0	6.0	6.2	92.0	76.0	72.0	72.0	73.0
1.8	7.7	5.9	5.8	5.4	5.9	93.0	68.0	68.0	66.0	70.0
3.2	7.6	6.4	6.2	5.5	5.6	91.0	76.0	73.0	67.0	68.0
5.6	7.7	6.4	6.8			92.0	79.0	81.0
10	7.6	6.8				92.0	82.0
	pH					Temperature (°C)
Control	8.1	8.1	8.1	8.1	8.0	25	24	24	24	23
1	7.9	8.2	8.1	8.1	8.0	25	24	24	24	23
1.8	8.0	8.2	8.1	8.0	8.0	25	24	24	24	24
3.2	8.0	8.2	8.1	8.0	8.0	25	24	24	24	23
5.6	8.1	8.2	8.1			25	24	23
10	8.1	8.0				25	24

Conclusions:

LC50 (96h): 2.5 mg/l (graphically determined)

Executive summary:

The acute toxicity potential on fish was investigated following the procedures outlined into the OECD guideline 203. The test solutions were prepared by adding appropriate amounts of a stock solution of the test material to test water. The experiment was performed at the following nominal concentrations: 1, 1.8, 3.2, 5.6 and 10 mg/l. During the exposure period of 96 hours, the test item concentrations were measured at the start and the end of the exposure period and the geometrical mean vales resulted to be 0.7, 1.2, 2.1, 4.1 and 8.2 mg/l.

The maximum concentration producing no mortality resulted to be 1.65 mg/l, while the minimum concentration producing 100 % mortality was 3.5 mg/l. The cumulative percentage mortality was plotted against the concetration on logarithmic-probability paper. A line was fitted and the concentration corresponding to the 50 per cent response read off; the LD50 value resulted to be 2.5 mg/l.

Conclusion

LC50 (96h): 2.5 mg/l (graphically determined)

Description of key information

LC50 (96h): 2.5 mg/l (graphically determined)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

2.5 mg/L

Additional information

The acute toxicity potential on fish was investigated following the procedures outlined into the OECD guideline 203. Two studies were available, on different test samples.

In the key study, a test sample of high dyestuff content (ca. 94 %) was used. The experiment was run at the following nominal concentrations: 1, 1.8, 3.2, 5.6 and 10 mg/l. During the exposure period of 96 hours, the test item concentrations were measured at the start and the end of the exposure period and the geometrical mean vales resulted to be 0.7, 1.2, 2.1, 4.1 and 8.2 mg/l.

The maximum concentration producing no mortality resulted to be 1.65 mg/l, while the minimum concentration producing 100 % mortality was 3.5 mg/l. The LC50 value was determined to be 2.5 mg/l.

In the supporting study, a liquid test sample was used, with a dyestuff content of 17 %. Test concentrations were 10,18, 32, 56, 100 mg/l. The maximum concentration producing no mortality resulted to be 10 mg/l, while the minimum concentration producing 100 % mortality was 32 mg/l. The LC50 value was determined to be 22 mg/l, equivalent to 3.74 mg/l as active ingredient.

In a Japanese study from 1991, acute toxicity to fish (Oryzias Latipes) was investigated in a preliminary test, before a bioaccumulation test. After a 48 -hour semistatic exposure to test substance, a LC50 value of 6.81 mg/l was found. No details on test method and results are available; however, such result is consistent with other experimental evidences in fish.

Overall, all available data indicates toxicity of test substance to fish.