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

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

5-10-2017 to 7-10-2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed using standard test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Principles of method if other than guideline:

Short term toxicity of test chemical to aquatic invertebrates was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system.

GLP compliance:

no

Analytical monitoring:

no

Details on test solutions:

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.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Straus
- Source: Own breeding at University of Chemistry and Technology, Prague
- Age at study initiation (mean and range, SD): The animals used for the test shall be less than 24 h old and should not be first brood progeny
- Feeding during test: No feeding

ACCLIMATION - No data available

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

± 1 hr

Test temperature:

20±1°C

pH:

sample at concentration 200 mg/l: pH = 7.6 did not change during the test,
control 1: pH = 7.6 changed to pH = 7.4 during the test,
control 2: pH = 7.7 did not change during the test,
control 3: pH = 7.7 changed to pH = 7.6 during the test

Dissolved oxygen:

higher than 7.9 mg/L at the end of test

Nominal and measured concentrations:

10, 25 , 50 , 100 , 200 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 ml glass vessel
- fill volume: 25 ml
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Natural water (surface or ground water), reconstituted water or dechlorinated tap water are acceptable as culturing and dilution water if D. magna survives in it for the duration of the culturing, acclimation and testing without showing signs of stress. Waters in the range pH 6 to pH 9, with hardness between 140 mg/l and 275 mg/l (as CaCO3) are recommended.
As an example, the preparation of dilution water meeting the requirements is described below.
Dissolve known quantities of reagents in water. The dilution water prepared shall have a pH of 7.8 ± 0.5, a hardness of (225 ± 50) mg/l (expressed as CaCO3), a molar Ca + Mg ratio close to 4 + 1 and a dissolved oxygen concentration above 7 mg/l.

Prepare the solutions specified below:
- Calcium chloride solution: Dissolve 117.6 g of calcium chloride dihydrate (CaCl2.2H2O) in water (4.2) and make up to 1 l with water (4.2).
- Magnesium sulfate solution: Dissolve 49.3 g of magnesium sulfate heptahydrate (MgSO4.7H2O) in water (4.2) and make up to 1 l with water (4.2).
- Sodium bicarbonate solution: Dissolve 25.9 g of sodium bicarbonate (NaHCO3) in water (4.2) and make up to 1 l with water (4.2).
- Potassium chloride solution: Dissolve 2.3 g of potassium chloride (KCI) in water (4.2) and make up to 1 l with water (4.2).

Mixing
Mix 2.5 ml of each of the four solutions and make up to 1 l with water.
The dilution water shall be aerated until the dissolved oxygen concentration has reached saturation and the pH has stabilized. If necessary, adjust the pH to 7.8 ± 0.5 by adding sodium hydroxide (NaOH) solution or hydrochloric acid (HCI). The dilution water prepared in this way shall not be further aerated before use.

- Sodium hydroxide solution, e.g. [NaOH] : 1 mol/l.
- Hydrochloric acid, e.g. [HCl] : 1 mol/l.

Reference substance:
Dissolve 600 mg of potassium dichromate (K2Cr2O7) in water and make up to 1 l with water (4.2).

OTHER TEST CONDITIONS
- Adjustment of pH: no adjustment done
- Photoperiod: No - Darkness
- Light intensity:

CALCULATION:
EC50 was calculated using non linear regression by the software Prism 4.0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7)

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI 213.7 - 1171 mg/l

Results with reference substance (positive control):

- Results with reference substance valid
- EC50: 0.79 mg/L (24 hours)

Reported statistics and error estimates:

EC50 was calculated using non linear regression by the software Prism 4.0 OR (The differences in means of control and sample were estimated by the t-test for independent groups at a 95 % confidence level, all individual replicates were used (STATISTICA CZ – data analysis software system, version 9.0, StatSoft, Inc.). Statistically significant differences are for p < 0.05.)

Validity criteria fulfilled:

yes

Conclusions:

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.

Executive summary:

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/l concentrations 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.

Description of key information

Short term toxicity to aquatic invertebrate:

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.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

500 mg/L

Additional information

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.