Disodium 2-(2,4,5,7-tetrabromo-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)

Principles of method if other than guideline:

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).

GLP compliance:

no

Analytical monitoring:

not required

Remarks:

According to test (If substance is stable and if not analytical done by spectrophotometer)

Vehicle:

not specified

Details on test solutions:

The test substance was soluble in water. Therefore, the stock solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous 1 hour stirring for achieving test concentrations of 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/L ,respectively.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name:Zebra Fish
- Strain:Danio rerio
- Source: Unique Aqua Fish Corner
- Length at study initiation (length definition, mean, range and SD): average 1.78 cm
- Weight at study initiation (mean and range, SD): average 0.080g
-Housing:The fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration.
-Duration of housing: 15 days
-Photoperiod: 16 hours light and 8 hours dark
-Diet : Standard brand feed, once daily
-Acclimatization: seven days
-feeding frequency: Once in a day

Test type:

static

Water media type:

freshwater

Total exposure duration:

96 h

Hardness:

150.5 mg of CaCO3

Test temperature:

23 °C

pH:

7.1

Dissolved oxygen:

7.0 mg/l

Nominal and measured concentrations:

6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Plastic 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: 16 hours light and 8 hours dark

Reference substance (positive control):

not specified

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:

Sublethal observations / clinical signs:

8.1. Mortality and symptoms

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

Visible Symptoms:

Nominal Concentration (mg/l)

Number of fish

symptom (cumulated) after

3h

6h

24 h

48 h

72 h

96 h

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

2. pH and oxygen content:

		Number of	pH readings after
Nominal	fish		24h		48h		72h		96h
Concentration			V1	V2	V1	V2	V1	V2	V1	V2
(mg/l)
Control	4	4	7.3	7.3	7.4	7.4	7.5	7.5	7.7	7.7
6.25	4	4	7.5	7.5	7.6	7.6	7.7	7.7	8.0	8.0
12.5	4	4	7.5	7.5	7.6	7.6	7.9	7.9	8.0	8.0
25	4	4	7.6	7.6	7.7	7.7	7.9	7.9	8.0	8.0
50	4	4	7.7	7.7	7.8	7.8	8.1	8.1	8.2	8.2
100	4	4	7.9	7.9	8.0	8.0	8.1	8.1	8.2	8.2

Nominal	Number of fish		Oxygen content (mg/L) after
Concentration			24h		48h		72h		96h
(mg/l)			V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	6.9	6.9	6.8	6.8	6.7	6.7	6.6	6.6
6.25	4	4	6.8	6.8	6.7	6.7	6.6	6.6	6.5	6.5
12.5	4	4	6.8	6.8	6.7	6.7	6.6	6.6	6.5	6.5
25	4	4	6.8	6.8	6.7	6.7	6.5	6.5	6.4	6.4
50	4	4	6.8	6.8	6.6	6.6	6.5	6.5	6.4	6.4
100	4	4	6.8	6.8	6.6	6.6	6.5	6.5	6.4	6.4

8.3. Test temperature

Nominal	Number of fish		Temperature (°C) after
Concentration			24h		48h		72h		96h
(mg/l)			V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	23.9	23.9	24.1	24.1	24.2	24.2	24.1	24.1
6.25	4	4	23.9	23.9	24.1	24.1	24.2	24.2	24.1	24.1
12.5	4	4	23.9	23.9	24.1	24.1	24.2	24.2	24.1	24.1
25	4	4	23.9	23.9	24.1	24.1	24.2	24.2	24.1	24.1
50	4	4	23.9	23.9	24.1	24.1	24.2	24.2	24.1	24.1
100	4	4	23.9	23.9	24.1	24.1	24.2	24.2	24.1	24.1

Validity criteria fulfilled:

yes

Conclusions:

Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for test material on Zebra Fish Danio rerio was determine to be >100 mg/l.

Executive summary:

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was soluble in water. Therefore, the stock solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous 1 hour stirring for achieving test concentrations of 6.25mg/L,12.5mg/L, 25mg/L,50mg/L, 100mg/L , respectively and Zebra FishDanio rerio were exposed to these concentration for 96 hours.

Bowl 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 to test item to nominal test concentrations, LC50 was determine to be >100 mg/l . Based on the LC50, it can be consider that the chemical was not toxic and can be consider to be not classified as per the CLP classification criteria.

Description of key information

Short term toxicity to fish:

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).The test substance was soluble in water. Therefore, the stock solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous 1 hour stirring for achieving test concentrations of 6.25mg/L,12.5mg/L, 25mg/L,50mg/L, 100mg/L , respectivelyand Zebra FishDanio reriowere exposed to these concentration for 96 hours.

Bowl 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 to test item to nominal test concentrations, LC50 was determine to be >100 mg/l . Based on the LC50, it can be consider that the chemical was not toxic 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

Short term toxicity to fish:

The short term toxicity to fish for the test material was determined based on the data derived from experimental report which was further supported by data from peer reviewed journal.

The experimental study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).The test substance was soluble in water. Therefore, the stock solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous 1 hour stirring for achieving test concentrations of 6.25mg/L,12.5mg/L, 25mg/L,50mg/L, 100mg/L , respectivelyand Zebra FishDanio reriowere exposed to these concentration for 96 hours.

Bowl 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 to test item to nominal test concentrations, LC50 was determine to be >100 mg/l . Based on the LC50, it can be consider that the chemical was not toxic and can be consider to be not classified as per the CLP classification criteria. 

The above experimental data was supported by data in peer reviewed journal In which short term toxicity study to Gambusia Affiniswas carried out for 96 hrs. The concentration of test chemical used for the study was 1, 10, 20, 50, 70, 100, 150 and 200 mg/l, respectively. The fish Gambusia Affinis Baird and Girard, were collected from a pond at Mississippi State University.

 

The test was performed under static conditions for 96 hrs. The fishes were kept in the laboratory for at least 72 hrs for stabilization prior to their use in the test. 20 fishes were then transferred into a 5 gallon glass aquarium after the appropriate amount of dyes was added. The aquarium were illuminated with Cool white Rflorescent lamps giving a surface light intensity of 3800 µW/cm2.Mortality was recorded as a function of illumination time and dye concentration.

 

The criterion for the death of fish was lack of movement immediately after touching with a camel hair brush. The light intensity was measured with an EG & GRphotometer/radiometer, Model 450-1. Abbott’s formula was used to correct for mortality in controls. Probit analysis was used to determine the log-time mortality regression lines and to calculate LC50 values.As no mortality of test organism was observed, the 96 hr LC50 value was found to be > 200 mg/l, respectively.

The above experimental data was further supported by Short term toxicity study to Oryzias latipes was carried out for 48 hrs and survival rate (in %) was determined. Himedaka (Oryzias latipes) of same age (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 litre solution of pH 7.0 containing 3,000 mg/l of test substance Eosine, ten fishes were kept in the tank without direct sunlight for 48 hrs and survival rate was determined. Water temp. was 20ᵒC and aeration was provided with bubbler.

 Median Tolerance Limit (TLm) was also determined after 24 and 48 hrs.After observation of dead fish, coloration of gill, fin and mouth by the test substance Eosine was recognized and particularly deep coloration of gill was found. Though internal organs were not colored, the excrement was clearly colored .Significant binding of Eosine to bovine serum albumin was also noted.The TLm value was found to be 2,700 and 2,200 mg/l at 24 and 48 hrs.Based on mortality of test fishes, the LC50 value was found to be 2,700 and 2,200 mg/l at 24 and 48 hrs, respectively and the 48 hr LC100 value was found to be 3,000 mg/l.

In another peer reviewed journal , Short term toxicity study to Oryzias latipes was carried out for 48 hrs and lethal concentration of 50% fish was determined. Himedaka (Oryzias latipes) of same age (2 cm long and 0.2 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.

Median Tolerance Limit (TLm) was determined after 24 and 48 hrs. The TLm value was found to be 1,000 and 620 mg/l at 24 and 48 hrs. Based on mortality of test fishes, the LC50 value was found to be 1,000 and 620 mg/l at 24 and 48 hrs, respectively.

In another journal , In a short-term toxicity to fish study,test material was exposed toOryzias latipes  for 48 hours with or without oxygen. Results shows that number of dead fish increased remarkably by irradiation of test material and all fish died after 7-711 hours. While under the bubbling of oxygen gas, the time of death was delayed and all the fish died after 16-24 hours. Therefore, LC100 was considered to be 3000 mg/l for test material when Oryzias latipes were exposed for 48 hours.

One of the peer reviewed journal short term toxicity study to Oryzias latipes was carried out for 48 hrs and TLm value was determined. Himedaka (Oryzias latipes) of same age (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 litre solution of pH 7.0 containing 3,000 mg/l of test substance test material, ten fishes were kept in the tank without direct sunlight for 48 hrs and survival rate was determined. Water temp. was 25ᵒC and aeration was provided with bubbler. Median Tolerance Limit (TLm) was determined after 48 hrs. The TLm value was found to be 1,800 mg/lat 48 hrs. Based on mortality of test fishes, the 48 hr LC50 value was found to be 1,800 mg/l.

Based on the above effect concentrations determined from experimental data and data from all the peer reviewed journals , it can be concluded that test material is not toxic to fish and can be considered to be not classified as per CLP criteria.