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REACH

3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid

EC number: 242-355-6 | CAS number: 18472-87-2

Life Cycle description
Uses advised against

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

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 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid
(CAS No. 18472-87-2) on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).

GLP compliance:

not specified

Analytical monitoring:

not required

Vehicle:

not specified

Details on test solutions:

The test substance was highly soluble in water,In house solubility was found to be 786 g/L. Therefore the stock solution prepared as 1g/1L, with the concentration of 1g/1L, and was kept for 5 min. stirring. After this as it is stock taken for experiment. From this stock solution further test concentrations were prepared for achieving 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:

TEST ORGANISM
- Common name: Zebra fish
- Source: Unique Aqua Fish Corner
- Length at study initiation (length definition, mean, range and SD): average 2.32cm
- Weight at study initiation (mean and range, SD): Average 0.3038 gm
- 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
- Physical conditions: No abnormalities
- Method of breeding: No data
- Maintenance of the brood fish:No data

ACCLIMATION
- Acclimation period:7 days
- Acclimation conditions: Photoperiod:12 hours light and 12hour darkness, Temperature range: 24.6°C, pH range: 7.315, DO range: 7.15mg/L.
- Type and amount of food during acclimation:Standard brand feed
- Feeding frequency during acclimation: Once in a day up to sixth day of acclimatization
- Health during acclimation (any mortality observed): During last 48 hours = 0%

FEEDING DURING TEST
- Food type:Standard brand feed
- Frequency:once daily
- Withdrawal of food: 1 day before and during exposure

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

151.2 mg of CaCO3

Test temperature:

23°C

pH:

7.02

Dissolved oxygen:

7.15 mg/L

Salinity:

No data available

Conductivity:

No data available

Nominal and measured concentrations:

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

Details on test conditions:

TEST SYSTEM
- Test vessel:The test vessels used were all plastic aquaria having the water capacity of 5liters.
- Material, size, headspace, fill volume: 2 liters
- Aeration: Continous (Aeration in test vessels was provided 1 day before the start of experiment.)
- No. of organisms per vessel:8 fishes
- No. of vessels per concentration (replicates): No data available
- No. of vessels per control (replicates): No data available
- No. of vessels per vehicle control (replicates): No data available
- Biomass loading rate: No data available

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod:12 hours light and 12 hours darkness
- Light intensity: No data available

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :Determination or calculation of the median lethal concentrations (LC50) and observations (mortality, visible symptoms, pH, Temperature, dissolved oxygen content) were recorded after 24, 48, 72 and 96 hours of the start of the experiment.

TEST CONCENTRATIONS
- Test concentrations: 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L, 100 mg/L .

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)

Sublethal observations / clinical signs:

Mortality and symptoms

NominalConcentration (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	-	-	-	-	-		-	-	-	-	-	-	-

Visible Symptoms:

NominalConcentration

(mg/l)

Number of fish

Symptoms after

3 h

6 h

24h

48h

72h

96h

0 (control)

NS4

6.25

NS4

12.5

NS4

100

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:Swimmingbehaviour

RF:Respiratory Function

P:Pigmentation

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	8.30		7.57	7.71	7.72	7.84	8.09	7.92	7.77
6.25	4	4	7.95		8.00	7.90	7.81	8.01	8.02	7.99	8.00
12.5	4	4	8.04		8.03	7.78	7.90	8.13	8.07	7.98	7.76
25	4	4	8.07		7.89	7.99	8.00	8.09	8.09	8.15	8.11
50	4	4	8.01		8.08	8.00	8.01	8.14	8.08	7.89	7.82
100	4	4	8.17		8.16	7.98	7.99	8.08	8.07	7.76	7.80

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	7.7		7.4	8.1	8.2	8.5	8.5	8.6	8.7
6.25	4	4	7.4		7.5	8.3	8.3	8.4	8.4	8.6	8.6
12.5	4	4	7.5		7.3	8.2	8.2	8.4	8.3	8.69	8.7
25	4	4	7.3		7.2	8.2	8.3	8.5	8.5	8.7	8.6
50	4	4	7.3		7.3	8.3	8.2	8.4	8.4	8.6	8.7
100	4	4	7.2		7.3	8.1	8.2	8.4	8.4	8.7	8.6

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	22.7		22.7	23.5	23.2	21.1	21.1	21.4	21.2
6.25	4	4	22.5		22.8	23.3	23.4	21.0	21.3	21.2	21.1
12.5	4	4	22.6		22.5	23.4	22.7	21.0	21.0	21.1	20.8
25	4	4	22.4		22.3	22.6	22.7	20.9	20.9	20.8	20.8
50	4	4	22.1		22.1	22.2	22.3	20.7	20.6	20.8	20.4
100	4	4	22.0		22.0	22.1	22.0	20.6	20.6	20.4	20.6

Validity criteria fulfilled:

yes

Conclusions:

Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for CAS No. 18472-87-2 on Zebra fish Danio rerio, was determined to be >100 mg/L

Executive summary:

The study was conducted to access the effect of test chemical 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen -9-yl)benzoic acid (CAS No. 18472) on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was highly soluble in water. The stock solution was prepared as 1g/1L with a concentration of 1g/1L and was kept for 5 min stirring. From this stock solution, further test concentrations were prepared for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L, and 100 mg/L respectively. This test solution was then added to the remaining two liters of water for achieving test concentrations of 100 mg/L and Zebra FishDanio reriowere exposed to these concentrations for 96 hours. Bowl aquaria containing 2 liters of potable water (passed through a reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with OECD guideline 203. After 96 hours of exposure to test substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen -9-yl)benzoic acid to various nominal test concentrations, LC50 was determine to be >100 mg/l . Based on the LC50 and because of the substance is readily biodegradable, it can be concluded that the chemical was non-hazardous and can be considered to be not classified as per the CLP classification criteria.

Description of key information

Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for CAS No. 18472-87-2 on Zebra fish Danio rerio, was determined to be >100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Effect concentration:: 100 mg/L

Additional information

Various studies including experimental data following OECD guideline for toxicity to aquatic fish with the target substance 3,4,5,6-tetrachloro-2-(1,4,5,8- tetra bromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2), were summarized as follows:

Study was conducted to access the effect of test chemical 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen -9-yl)benzoic acid on the growth of fish Danio rerio. Test

conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was highly soluble in water.In house solubility was found to be 786 g/L. Therefore the stock solution prepared as 1g/1L, with the concentration of 1g/1L, and was kept for 5 min. stirring. After this as it is stock taken for experiment. From this stock solution further test concentrations were prepared for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L, 100 mg/L respectively.This test solution was then added to the remaining two liters of water for achieving test concentrations of 100 mg/L and 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 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen -9-yl)benzoic acid to various nominal test concentrations, LC50 was determine to be >100 mg/l . Based on the LC50, it can be consider that the chemical was non hazardous and can be consider to be not classified as per the CLP classification criteria.

Moreover, the study was conducted by Pimprikar et al. (Southwest. Entomol.9 (2): 218-222 1984) to determine the toxic effects of 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472) to predatory mosquitofish. The toxicity was observed over a period of 4 days. The lethal concentrations (LC50) for the test substance, 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo- 6-hydroxy-3-oxoxanthen-9-yl)benzoic acid, in Gambusia affinis on a mortality effect was observed to be 97.7 mg/L. Also the substance was found to be not rapidly degradable. Thus according to CLP Criteria for aquatic classification of the substance, it is concluded that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6- hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS No. 18472-87-2) exhibits toxicity to aquatic fish in the chronic 3 classification category.

Futhermore,Short term toxicity study toOryzias latipeswas carried out for 48 hrs and survival rate (in %) & Median Tolerance Limit (TLm) 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 Phloxine B, ten fishes were kept in the tank without direct sunlight for 48 hrs and survival rate was determined. Water tem. was 20ᵒC and aeration was provided with bubbler.Median Tolerance Limit (TLm) was also determined after 24 and 48 hrs.After observation of dead fishes, coloration of gill, fin and mouth by the test substance Phloxine B was recognized and particularly deep coloration of gill was found. Though internal organs were not colored, the excrement was clearly colored. Significant binding of Phloxine B to bovine serum albumin was also noted.The TLm value was found to be 230 and 190 mg/l at 24 and 48 hrs.Based on mortality of all test fishes, the LC100 value was determined to be 230 and 190 mg/l at 24 and 48 hrs respectively. Thus, the survival rate of fish after 48 hrs was determined to be 0%.

So, based on the above studies for substance 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3-oxoxanthen-9-yl)benzoic acid (CAS Number: 18472-87-2), the lethal concentration LC50 was found to be in the range of 97.7 mg/L to >100.0 mg/L in a 96 hours of exposure. Although data from peer-reviewed journal suggests that the substance is classified in aquatic chronic 3 but as the data from experimental study has K1 reliability and the test was performed using OECD guidelines which has the value of LC50 as >100 mg/L in a 96 hours of study, indicating that 3,4,5,6-tetrachloro-2-(1,4,5,8-tetrabromo-6-hydroxy-3 -oxoxanthen -9-yl)benzoic acid (CAS No. 18472-87-2) does not exhibits toxicity to fish criteria of CLP, the same has been considered for the purpose of classification and chemical safety assessment.

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