Trisodium bis[3-hydroxy-4-[(2-hydroxy-1-naphthyl)azo]naphthalene-1-sulphonato(3-)]chromate(3-)

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:

September from 17 to 21, 1990

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:

other: DIN 38412

Version / remarks:

June 1982

GLP compliance:

no

Analytical monitoring:

no

Details on test solutions:

The product was added to the test water without any pre-treatment; subsequently the fish were placed into the aquaria.

Test organisms (species):

Leuciscus idus

Details on test organisms:

TEST ORGANISM
- Common name: golden orfe.
- Source: Fischzucht Paul Aggers, D-W2354 Hohenwestedt, FRG.
- Length at study initiation: 5.3 cm (4.2 - 6.0 cm).
- Weight at study initiation: 1.7g (0.8 - 2.4g).

ACCLIMATION and HOUSING
- Adaptation period: housing and adaptation last about 2 month
- Housing conditions: the fish were kept in a flow-through tank in tap water cleaned by active carbon and aerated with oil-free air.
Photoperiod: 16 hours light and 8 hours dark.
Total hardness: about 2.5 mmol/l
Acid capacity: about 5.5 mmol/l
Oxygen content: higher than 60 % of maximum saturation.
pH: about 8.0
Temperature: 20 - 24 °C
- Mortality during the last 2 weeks of housing: about 1.8 %
- Mortality during the last adaptation period: 1.7 %
- Medical treatment: twice with 0.05 mg/l malachite green chloride once with 10 mg/l tetracycline hydrochloride.
- Diet: growing feed F/B 50, Ssniff Spezialdiaeten GmbH, ad libitum.

ADAPTATION
- Adaptation: adaptation to test water and test temperature 3 days.

FEEDING DURING TEST
- Withdrawal of food: 1 day before and during exposure.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

Test vessels: 21 - 23 °C
Control: 22 - 23 °C

pH:

Test vessels: 7.8 - 8.0
Control: 7.8 - 8.1

Dissolved oxygen:

Test vessels: 7.7 - 8.7 mg/l
Control: 7.9 - 8.1 mg/l

Nominal and measured concentrations:

10.0, 21.5, 46.4 and 100 mg/l (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessels: all-glass aquarium (30 × 22 × 24 cm).
- Volume of water: 10 litres
- Aeration: slight; continuous with oil-free air.
- No. of organisms per concentration: 10 fish.
- Loading rate: 1.7 g fish/l test water.

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: reconstituted freshwater according to DIN 38 412, part 11, October 1982; preparation from fully demineralized tap water.
- Conductivity: max. 10 μƱ
- CaCl2 × 2 H2O: 294.0 mg/l
- MgSO4 × 7 H2O: 123.3 mg/l
- NaHCO3: 63.0 mg/l
- KCl: 5.5 mg/l
- Total hardness: 2.5 mmol/l
- Acid capacity: 0.8 mmol/l
- Ratio Ca/Mg: 4 : 1
- Ratio Na/K: 10 : 1
- pH: about 8.0

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light and 8 hours darkness.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Range finding study: LC50 (96h) resulted between 10 and 100 mg/l.

Reference substance (positive control):

yes

Remarks:

chloroacetamide

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

21.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No Observed Effect Concentration: 10.0 mg/l
Maximum concentration tested causing no mortality: 10.0 mg/l
Minimum concentration tested causing 100 % mortality: 46.4 mg/l

Results with reference substance (positive control):

Positive control of animals conducted with chloroacetamide LC50 (48h) about 32 mg/l; the letahl concentration corresponds to the normal sensitivity.

Sublethal observations / clinical signs:

Deaths and symptoms recorded

Nominal conc. (mg/l)	N. of fish	Dead fish after
		1 hr	24 hrs	48 hrs	72 hrs	96 hrs
10.5	10	0	0	0	0	0
21.5	10	0	5	5	5	6
46.4	10	1	10	10	10	10
100.0	10	10	10	10	10	10
0.0	10	0	0	0	0	0
Symptoms
10.5
21.5	U		(A)		T	T
46.4	A
100.0

A: apathy

T: tumbling

U: restlessness

(): slight to very slight

TEST CONDITIONS

Nominal conc. (mg/l)	pH
	1 hr	24 hrs	48 hrs	72 hrs	96 hrs
10.5	8.0	7.9	7.9	7.8	7.8
21.5	7.9	7.8	7.8	7.9	7.8
46.4	8.0	7.8
100.0	7.9
0.0	8.1	7.9	7.9	7.8	7.8
	Oxygen content (mg/l)
10.5	8.2	7.9	7.9	8.0	7.8
21.5	8.2	7.7	7.8	8.0	7.8
46.4	8.5	8.3
100.0	8.7
0.0	8.1	8.0	7.9	8.1	7.9
	Temperature (°C)
10.5	22	23	23	23	22
21.5	22	22	22	22	22
46.4	22	22
100.0	21
0.0	22	23	23	23	22

Conclusions:

LC50 (96h): 21.5 mg/l (nominal)

Executive summary:

The substance was tested for acute toxicity to fish following the testing procedures outlined into the DIN 38412. 10 fish were used for each concentration tested and exposed to the substance over a period of 96 hours. Based on a preliminary test, the following nominal concentrations were chosen for the main experiment: 0 (blank), 10.5, 21.5, 46.4 and 100.0 mg/l. No analytical determination of tested concentration was performed.

The No Observed Effect Concentration at 96 hours resulted to be 10.0 mg/l (nominal), as well as the maximum concentration tested causing no mortality. The minimum concentration tested causing 100 % mortality was 46.4 mg/l (nominal).

Conclusion

LC50 (96h): 21.5 mg/l (nominal)

Description of key information

LC50 (96h): 21.5 mg/l (nominal)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

21.5 mg/L

Additional information

Acid Blue 193 was tested for acute toxicity to fish following the testing procedures outlined into the DIN 38412. Based on a preliminary test, the following nominal concentrations were chosen for the main experiment: 0 (blank), 10.5, 21.5, 46.4 and 100.0 mg/l. No analytical determination of tested concentration was performed. The No Observed Effect Concentration at 96 hours resulted to be 10.0 mg/l (nominal), as well as the maximum concentration tested causing no mortality. The minimum concentration tested causing 100 % mortality was 46.4 mg/l (nominal).

It should be noted that, during housing and adaptation period lasted ca 2 months, fish were treated with a known toxic substance, i.e. Malachite Green. Fish were treated twice with 0.05 mg/l malachite green chloride once with 10 mg/l tetracycline hydrochloride. Malachite Green is listed in annex VI of the CLP Regulation (EC) 1272/2008 and classified as toxic for aquatic life: Aquatic Acute 1 (H400) and Aquatic Chronic 1 (H410).

Although a contribute on the long period cannot be excluded, the effects recorded were considered mainly due to the Acid Blue 193 treatment.

In addition, an sheet of testing results is availabe. No details about testing method, procedures and results are given, thus a reliability cannot be assigned. Anyway, results are consistent with those obtained into the key study. A LD50 (72h) of 31.6 mg/l was indicated for Brachydanio rerio (reference given in attachment).