Disodium 2-(2,4,5,7-tetraiodo-6-oxido-3-oxoxanthen-9-yl)benzoate

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed according to the guideline.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

The stock solution was prepared by dissolving 1 gm of the test substance in 1 liters of potable water (passed through reverse osmosis system). This stock solution was used for preparing test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L, respectively.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Animal species: Zebra Fish (Danio rerio)
Body length: Average 2.33 cm
Body weight: 0.127g (average)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

24, 48, 72 and 96 hours

Test temperature:

23.7-24.7°C

pH:

7.1

Dissolved oxygen:

8.3mg/l

Nominal and measured concentrations:

Nominal test concentrations: 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L, respectively

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquaria
- Material, size, headspace, fill volume: 5 liters of plastic aquaria filled with 2 liter of water
- Aeration: Aeration in test vessels was provided 1 day before the start of experiment
- No. of organisms per vessel: 8 fishes

OTHER TEST CONDITIONS
- Photoperiod: 12 hours light and 12-hour darkness

Water: Potable water (passed through reverse osmosis system) was used
pH: 7.1
Hardness: 53.75 (mg of CaCO3)
TDS: 382 ppm
Conductivity: 1072 (µScm-1)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No mortality were found

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: As no effects were observed on the fishes, thus LC50 value consider to be > 100 mg/l

Sublethal observations / clinical signs:

Table 1: Mortality and symptoms

Nominal Concentration (mg/l)	Number of fish		Mortality (cumulated) after
			3 h		6 h		24h			48h		72h		96h
			V1	V2	V1	V2	V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	-	-	-	-	-		-	-	-	-	-	-	-
6.25	4	4	-	-	-	-	-		-	-	-	-	-	-	-
12.5	4	4	-	-	-	-	-		-	-	-	-	-	-	-
25	4	4	-	-	-	-	-		-	-	-	-	-	-	-
50	4	4	-	-	-	-	-		-	-	-	-	-	-	-
100	4	4	-	-	-	-	-		-	-	-	-	-	-	-

Table 2: Symptoms

Nominal Concentration (mg/l)

Number of fish

Symptoms after

3 h

6 h

24h

48h

72h

96h

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

0 (control)

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

6.25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

12.5

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

50

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

100

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

Explanation of symptoms:

A:apathy

N:Narcotic-like state

C:Swimming near water surface

M: Slow movements compared to control

NS:No abnormal symptoms 

S:Surfacing behaviour

L:Loss of equilibrium

SM:Shimming movement

H1:Hyperactivity

H2:Hypo activity

B:Bending behavior

F:Fin rot

IS:Invert Swimming

SB:Swimming behaviour

RF:Respiratory Function

P:Pigmentation

Table 3: pH and oxygen content:

NominalConcentration (mg/l)	Number of fish		pH readings after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	8.5		8.5	8.3	8.4	8.2	8.3	8.3	8.4
6.25	4	4	8.5		8.4	8.5	8.5	8.4	8.4	8.5	8.4
12.5	4	4	8.5		8.4	8.4	8.4	8.4	8.4	8.4	8.5
25	4	4	8.5		8.5	8.5	8.3	8.5	8.3	8.5	8.4
50	4	4	8.5		8.5	8.3	8.4	8.3	8.3	8.3	8.3
100	4	4	8.4		8.6	8.3	8.5	8.3	8.5	8.3	8.5

NominalConcentration (mg/l)	Number of fish		Oxygen content (mg/L) after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	8.2		8.2	7.9	8.1	8.2	8.4	7.9	7.9
6.25	4	4	8.2		8.1	8.1	8.1	7.9	8.1	7.9	7.9
12.5	4	4	8.2		8.2	7.9	7.8	7.8	8.2	7.9	7.9
25	4	4	8.2		8.1	7.9	7.8	7.8	7.8	7.8	7.8
50	4	4	8.2		8.1	7.9	7.8	7.8	7.8	7.8	7.8
100	4	4	8.1		7.9	7.9	7.8	7.9	8.2	7.8	7.8

Table 4: Test temperature

NominalConcentration (mg/l)	Number of fish		Temperature (°C) after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	22.6		22.5	22.8	22.7	22.6	22.6	22.8	22.5
6.25	4	4	22.5		22.2	22.7	22.6	22.4	22.3	22.4	22.2
12.5	4	4	22.4		22.4	22.6	22.6	22.4	22.3	22.3	22.3
25	4	4	22.4		22.3	22.6	22.3	22.1	22.1	22.2	22.0
50	4	4	22.4		22.3	22.3	22.4	22.3	22.1	22.1	22.1
100	4	4	22.2		22.4	22.3	22.4	22.1	22.3	22.0	22.2

Validity criteria fulfilled:

yes

Conclusions:

Based on nominal concentrations, experimental median lethal Concentrations [LC-50] of test chemical on Zebra Fish (Denio rario) after exposing for 96 hours was determined to be > 100 mg/L, as no mortality were observed at 100 mg/l.

Executive summary:

Study was conducted to assess the effect of test chemical on the mortality of fishes. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). Test conducted on Danio rerio (previous name: Brachydanio rerio). The stock solution was prepared by dissolving 1 gm of the test substance in 1 liters of potable water (passed through reverse osmosis system). This stock solution was used for preparing test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L, respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure of test chemical to various nominal test concentrations, LC50 was calculated and the mortality was used as a key parameter for the determination of exact effect on fishes. Based on nominal concentrations, experimental median lethal Concentrations [LC-50] of test chemical on Zebra Fish (Denio rario) after exposing for 96 hours was determined to be > 100 mg/L, as no mortality were observed at 100 mg/l. Thus, it can be consider that the chemical was non hazardous and can be consider to be not classified as per the CLP classification criteria.

Description of key information

Study was conducted to assess the effect of test chemical on the mortality of fishes. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). Test conducted on Danio rerio (previous name: Brachydanio rerio). The stock solution was prepared by dissolving 1 gm of the test substance in 1 liters of potable water (passed through reverse osmosis system). This stock solution was used for preparing test concentrations of 6.25 mg/L, 12.5 mg/L, and 25 mg/L, 50 mg/L & 100 mg/L, respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure of test chemical to various nominal test concentrations, LC50 was calculated and the mortality was used as a key parameter for the determination of exact effect on fishes. Based on nominal concentrations, experimental median lethal Concentrations [LC-50] of test chemical on Zebra Fish (Denio rario) after exposing for 96 hours was determined to be > 100 mg/L, as no mortality were observed at 100 mg/l. Thus, it can be consider that the chemical was nonhazardous and can be consider to be not classified as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Summarized result from the various experimental sources for the determination of effect of test chemical on the mortality of fishes are as mentioned below:

 

Study was conducted to assess the effect of test chemical on the mortality of fishes. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). Test conducted on Danio rerio (previous name: Brachydanio rerio). The stock solution was prepared by dissolving 1 gm of the test substance in 1 liters of potable water (passed through reverse osmosis system). This stock solution was used for preparing test concentrations of 6.25 mg/L, 12.5 mg/L, and 25 mg/L, 50 mg/L & 100 mg/L, respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure of test chemical to various nominal test concentrations, LC50 was calculated and the mortality was used as a key parameter for the determination of exact effect on fishes. Based on nominal concentrations, experimental median lethal Concentrations [LC-50] of test chemical on Zebra Fish (Denio rario) after exposing for 96 hours was determined to be > 100 mg/L, as no mortality were observed at 100 mg/l. Thus, it can be consider that the chemical was nonhazardous and can be consider to be not classified as per the CLP classification criteria.

 

Above study further supported by the second study from peer reviewed journal. Principle of this study was to determine the effect of test chemical on the mortality rate of fishes. Gambusia affinis was used as a test organism and the study performed under the static system for 96 hours. Different test concentrations 1, 10,20,50,70,100,150 and 200 mg/l respectively were used. Fishes were collected from the pound and kept in the laboratory for at least 72 hr for the stabilization prior to their use in the tests. Fishes were starved for 24hr before test. 20 fishes were transferred to the 5 gallon glass aquaria were illuminated with fluorescent lamps giving surface light intensity of 3800 µw/cm2. Probit analysis was used to determine the effect of test chemical, whereas mortality were used as a key parameter for the LC50 calculation. After the exposure of test chemical for 96 hrs with fishes Gambusia affinis, the LC50 value was determine to be > 200 mg/l on the basis of mortality observations.

 

Short term toxicity test was performed on Himedaka (Oryzias latipes). All fishes acclimated for 10 days in tap water before experiment. In lab, 1 liter solution having pH 7 containing 3,000 mg/liter of test chemical to be tested. 10 tested fish were kept in the tank without direct sunlight for 48 hrs and survival rate was determined. It was observed that at the 3000 mg/liter concentration, the survival rate of fish was observed to be 0 % and thus the LC100 was consider to be at 3000 mg/l.

Similarly an acute study was conducted to determine the effect of test chemical on the fishes. Test conducted in accordance with the Japanese Industrial Standards Committee. Himedaka commonly known as Oryzias latipes of same age i.e. 2 cm long and 0.2 gram in weight was used for the study. The test fishes were acclimatized for 10 days in tap water. Ten fish of Himedaka per one trial were kept in 2 liter of deionized water at 25°C and, after 24 and 48 hrs, lethal concentration of 50% fish was determined. Based on the mortality of fishes Oryzias latipes due to the test chemical exposure for 24 and 48 hours, the LC50 value was determine to be 710 and 340 mg/l respectively.

 

The fifth study from peer reviewed journal was conducted on Oryzias latipes was carried out for 48 hrs and TLm value was determined. Himedaka (Oryzias latipes) of same age i.e. 2 cm long and 0.2 in weight was used for the study. The test fishes were acclimatized for 10 days in tap water. In 1 liter solution of pH 7.0 containing test substance, ten fishes were added in the tank without direct sunlight for 48 hrs and survival rate was determined. Water temperature was 25°C and aeration was provided with bubbler. Median Tolerance Limit (TLm) was determined after 48 hrs. Based on the mortality of test fishes, the 48 hrs LC50 value was determine to be 900 mg/l.

 

Thus, on the basis of above all studies and effects observations on fishes, test chemical consider to be nontoxic and not classified as per the CLP classification criteria.