Trisodium 8-hydroxypyrene-1,3,6-trisulphonate

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 test solution was prepared by dissolving 400 mg of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous stirring. This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L.

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 3.18 cm
- Weight at study initiation (mean and range, SD): average 0.180 g
-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 hour darkness
-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:

142.5 mg of CaCO3

Test temperature:

23 °C

pH:

7.98

Dissolved oxygen:

7.2 mg/l

Nominal and measured concentrations:

100 mg/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 darkness

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: No mortality was observed

Sublethal observations / clinical signs:

8.1. Mortality and symptoms:

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

Visible Symptoms:

Nominal Concentration (mg/l)

Number of fish

symptom (cumulated) after

3h

6h

24h

48h

72h

96h

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

Control

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

pH and oxygen content:

Nominal Concentration (mg/l)	Number of fish		pH readings after
			24h		48h		72h		96h
			V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	8.21	8.22	7.57	7.60	7.40	7.47	7.51	7.53
100	4	4	7.65	7.64	7.40	8.21	8.21	8.17	8.11	8.13

Nominal Concentration (mg/l)	Number of fish		Temperature (°C) after
			24h		48h		72h		96h
			V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	21	21	25	25	24	24	23	23
100	4	4	21	21	25	25	24	24	23	23

Validity criteria fulfilled:

yes

Conclusions:

Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for CAS No. 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 test solution was prepared by dissolving 400 mg of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous stirring. This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L. This test solution was then added to the remaining three 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 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 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 test solution was prepared by dissolving 400 mg of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous stirring. This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L.This test solution was then added to the remaining three 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 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 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:

Short term toxicity to fish was evaluate based on the experimental report and other peer reviewed journal.

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 test solution was prepared by dissolving 400 mg of the test substance in 1 liters of potable water (passed through reverse osmosis system) with continuous stirring. This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L.This test solution was then added to the remaining three 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 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 not toxic and can be consider to be not classified as per the CLP classification criteria.

In a data from peer reviewed journal,to study the effects of test material on aquatic invertebrate acute toxicity tofish test was carried out for 96.0 hr under static condition. These values are toxicity values, not exposure values. SAR results are generally regarded by EPA as being reliable. Dyes with aquatic toxicity values below 1mg/l received a HIGH concern rating, those with toxicity values ranging from 1-100mg/l received a MODERATE concern rating and those with toxicity values greater than 100mg/l received a LOW concern rating. The Median Lethal Concentration which is estimated to be lethal to 50% of fish in 96 hours exposure to the test chemical (LC50) was found to be greater than 500mg/l. Since the LC50 for the test chemical is greater than 100 mg/l , it falls in the LOW concern category in the aquatic toxicity ratings. It can be concluded from the value that the test material is not toxic to the aquatic invertebrate and can be considered as “not classified” as per the classification criteria for aquatic environment.

In another peer reviewed journal , ecotoxicological assessments were based on the determination of acute toxicity to daphniae according to DIN 38412, Part 11 (Deutsches Institut für Normung e.V. 1982), and acute toxicity to fish according to DIN 38412, Part 31 (Deutsches Institut für Normung e.V. 1989). For the fish test, zebra fish were used as a modification of the guideline.

The Median Lethal Concentration which is found to be lethal to 0% of zebrafish in a 96 hours exposure to the test chemical (LC0) was found to be greater than 10mg/l

The fluorescent dyes that showed no effect upon either the genotoxicity or the ecotoxicity tests were classified by the Working Group as SAFE for use in water tracing.Test materialis classified as safe for use in water tracing.It can be concluded that the test substance is not toxic to the aquatic invertebrateand can be considered as “not classified” as per the classification criteria for aquatic environment.

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.

Short term toxicity to aquatic invertebrate:

The short term toxicity to aquatic invertebrate was evalauted based on experimental report and peer reviewed journal.

Aim of this study was to assess the short term toxicity of test material to aquatic invertebrates daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system for the total exposure period of 48 hrs.

 The stock solution 200 mg/l was prepared by dissolving yellow green powder reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water.10, 25 , 50 , 100 , 200 mg/lconcentrations were used in the study. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0.

 The median effective concentration (EC50) for the test substance , in Daphnia magna was determined to be 500 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be non hazardous to aquatic invertebrate and cannot be classified as aquatic as per the CLP classification criteria.

In a peer reviewed journal,to study the efffects of test material on aquatic invertebrate test was carried for 48 hr.The concern levels identified by use of the SAR technique are expressed ad HIGH, MODERATE, LOW. These values are toxicity values, not exposure values. SAR results are generally regarded by EPA as being reliable.

Dyes with aquatic toxicity values below 1mg/l received a HIGH concern rating, those with toxicity values ranging from 1-100mg/l received a MODERATE concern rating and those with toxicity values greater than 100mg/l received a LOW concern rating. The Median Lethal Concentration which is estimated to be lethal to 50% of Cladocera (water flea) in 48 hours exposure to the test chemical (LC50) was found to be greater than 100mg/l.

Since the LC50 for the test materialis greater than 100 mg/l, it falls in the LOW concern category in the aquatic toxicity ratings. Based on the result, thetest substancewas considered as "not classified" according to the CLP regulations.

In another peer reviewed journal,toxicity tests on the dyes were carried out using standard bioassay using the copepod acute toxicity test (Tisbe battagliai 48 hr LC50).Stock solutions of test dyes were prepared using filtered aerated seawater (0.2 μm). Serial dilutions were then prepared from the main stock to produce the test concentration range. Initial tests, or range finders, were carried out on bothTisbebattagliai. The range finder tests covered a concentration range of 0.001 to 1000 mg/L, where needed this was increased to 4000mg/L for tracer dye test material . The observations allowed for a narrower range to be achieved for more focused definitive studies.The T.battagliai bioassay followed the guidelines in ISO (1999) with no further modifications. These bioassays were carried out in 12 well polystyrene plates, each well contained 5mls of test solution and 5T.battagliaicopepodites (approximately 4-6 days old). There were 4 replicates per concentration and the test was carried out over a period of 48 hours. The Tisbe were observed using a binocular microscope and mortality recorded. Water quality checks for temperature, salinity, dissolved oxygen and pH were carried out at the start and the end of each test to ensure that physical parameters remained within the acceptable range, as set out in the test protocols, for each bioassay. All parameters were within acceptable limits, e.g. temperature (21°C ± 3°C), salinity (32 ± 2 %), dissolved oxygen (³80%) and pH (7.8 - 8.2) forT.battagliai bioassay.

 A 48 h Zinc reference study was carried for quality assurance purposes alongside the tracer dyes to test the sensitivity of the species population for validity of the study. This test uses several concentrations of Zinc sulphate 0 (control), 0.1, 0.18, 0.32, 0.56, 1.0 and 1.8 mg/L forT.battagliai bioassay. Atest is considered valid if the mortality ranges fall with the expected limits, i.e no more than 10% mortality forT.battagliai.

 The results showed lower levels of toxicity for test material for the test species, however, it is difficult to rule out whether these are considered to be safe for the marine environment.The LC50 for test material was 2703.1 mg/l. Test material was the least toxic tracer dye among the tracers tested. Based on the result, the chemical was considered as not classified for short term toxicity to aquatic invertebrates according to the CLP regulation.

Based on the EC50 value, substance is likely to be non hazardous to aquatic invertebrate and cannot be classified as aquatic as per the CLP classification criteria.