Reaction products of Hydrogen amino-4-hydroxynaphthalene-2-sulfonate, Potassium substituted-5-{[2-(sulfonatooxy)ethyl]sulfonyl}benzenesulfonate and Sodium substituted-({4-[(2-chloroethyl)sulfonyl]butanoyl}amino)benzenesulfonate

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

Study period:

2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Test item: FAT 40875/A TE
Physical appearance: Dark red powder
Purity: 86.4% all organic components
Batch No: BOP 05-17 (BS-ROE 1550 NIM 25+26+27)
Manufactured date: 20 December 2017
Expiry date: 11 December 2022
pH: 4.3 (aq. soln. 2 %w/w at room-temperature)
Recommended storage condition: Refrigeration (+2 to +8 °C) and protection from light.

Analytical monitoring:

yes

Details on sampling:

Sampling was done at the start and end of the test.

Vehicle:

yes

Details on test solutions:

Range Finding Test:
To prepare test concentrations, the test item (after grinding with mortar and pestle) was mixed with test medium (with portion of test medium from this group) along with sonication for 15 minutes followed by introduction into test aquarium containing 10 L of test medium. Stock (Stock-I - 1 mg/mL) solution was used for preparation of G2 and G3 test medium.

G1 (Control): 10 L of test medium in a test aquarium.
G2 (0.01 mg/L) : 100 μl of stock-I was added to test aquarium containing 10 L test medium.
G3 (0.1 mg/L): 1 mL of stock-I was added to test aquarium containing 10 L test medium.
G4 (1 mg/L): 10 mg test item was added to test aquarium containing 10 L test medium (15 mL of Stock-II – 0.67 mg/mL was used on day 0).
G5 (10 mg/L): 100 mg test item was added to test aquarium containing 10 L test medium.
G6 (50 mg/L): 500 mg test item was added to test aquarium containing 10 L test medium.
G7 (100 mg/L): 1000 mg test item was added to test aquarium containing 10 L test medium.

Definitive Test:
To prepare test concentration of 100 mg/L, 1000 mg of test item (after grinding with mortar and pestle) was mixed with test medium (with portion of test medium from this group) along with sonication for 15 minutes followed by introduction into test aquarium containing 10 L of test medium.
For control (G1), test aquarium containing 10 L of test medium was used.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: Tropical Fish, Shivajinagar, Bengaluru, India.
- Length at study initiation: 1 to 2 cm
- Weight at study initiation: 1.1023 g (Total body weight of ten fish)

ACCLIMATION
- Acclimation period: 7 days.
- Health during acclimation (any mortality observed): No mortality.
- Feeding: 3 times per week

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

250 as mg CaCO3/L

Test temperature:

22.2 to 22.7 °C

pH:

7.25 to 7.79

Dissolved oxygen:

87 to 96 %

Conductivity:

Less than 10 μS/cm.

Nominal and measured concentrations:

Definitive test: 0 (control), 100 mg/L

Details on test conditions:

TEST SYSTEM
- Renewal rate of test solution: test medium renewal at 24 h.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Water Deionizing unit (Milli-Q), Millipore India Pvt Ltd, Bengaluru, India.
- Total organic carbon: 0.8 mg/L

OTHER TEST CONDITIONS
- Photoperiod: 16 h photoperiod daily
- Light intensity: Range finding test - 612 to 640 lux, Definitive test - 590 lux.

EFFECT PARAMETERS MEASURED: Mortality.

TEST CONCENTRATIONS
- Range finding study : Yes
- Test concentrations: 0 (control), 0.01, 0.1, 1, 10, 50 and 100 mg/L
- Results used to determine the conditions for the definitive study: No mortality was observed at 100 mg/L in range finding test.

Reference substance (positive control):

no

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)

Details on results:

- Behavioural abnormalities: none observed.
- Observations on body length and weight: No abnormality detected in body weight and length.
- Other biological observations: none
- Mortality of control: no mortality observed in control.
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The test medium was found to be solution (red in colour) with the absence of settable solids at the test concentration of 100 mg/L.
- Effect concentrations exceeding solubility of substance in test medium: none.

Reported statistics and error estimates:

There was no mortality of fish observed in the tested concentration. Hence, no statistical analysis was employed to determine the LC50 of test item.

Sublethal observations / clinical signs:

Test Item Solubility: Test item was tested for its solubility in the test medium and formed red solution (along with sonication) with the test medium at 0.67 and 1 mg/mL (performed as a non-GLP test).

Test Item Stability in Test Medium and Analysis of Test Concentrations: The stability test results concluded that the test item was found to be stable in the test medium at room temperature up to 48 h at 2.02 and 101.0 mg/L of nominal concentrations.

The results of active ingredient analysis of 100 mg/L showed that overall percent agreement was 103.4% at the start of test and 100.40% at the end of test (48 h) indicating that the results were within the acceptable limit (80 to 120 % of the nominal concentration with an RSD of ≤ 20%). No measurable concentration of test item was recorded in the control samples.

Validity criteria fulfilled:

yes

Conclusions:

The NOEC (No Observed Effect Concentration) and LC50 for FAT 40875/A TE at 96 hours was higher than 100 mg test item/L based on nominal concentration.

Executive summary:

The acute toxicity of FAT 40875/A TE was tested on the freshwater fish, Danio rerio (Zebrafish). This test was conducted in accordance with OECD test guideline 203 in a GLP certified laboratory.

Fish were held in the laboratory at least for 12 days before using it for test. Healthy fish of length ranging from 1 to 2 cm were used in the test.

The range finding test was carried with the test concentrations of 0.01, 0.1, 1, 10, 50 and 100 mg/L along with control in a semi-static mode. Five fish per test solution were used in the test. No mortality or adverse effects were observed in the range finding test.

Based on the results of the range finding test, the definitive test was carried out using semi-static method at the test concentration of 100 mg/L along with a control. Seven fish were used for each test concentration.

Mortalities and visible abnormalities in regard to equilibrium, behavior and swimming behaviour were recorded. Test item stability in test medium and analysis of test concentration was also carried out similtaneously.

The stability test results concluded that the test item was found to be stable in the test medium at room temperature up to 48 h at 2.02 and 101.0 mg/L of nominal concentrations.

The results of active ingredient analysis of 100 mg/L showed that overall percent agreement was 103.4% at the start of test and 100.40% at the end of test (48 h) indicating that the results were within the acceptable limit (80 to 120 % of the nominal concentration with an RSD of ≤ 20%). No measurable concentration of test item was recorded in the control samples.

There were no mortality and toxic signs observed in control group as well as test item treated group of 100 mg/L up to 96 h exposure. The test medium was found to be solution (red in colour) with the absence of settable solids at the test concentration of 100 mg/L.

In conclusion, the NOEC (No Observed Effect Concentration) and LC50 for FAT 40875/A TE at 96 hours was higher than 100 mg test item/L based on nominal concentration.

Description of key information

The NOEC (No Observed Effect Concentration) and LC50 for FAT 40875/A TE at 96 hours was higher than 100 mg test item/L based on nominal concentration.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The acute toxicity of FAT 40875/A TE was tested on the freshwater fish, Danio rerio (Zebrafish). This test was conducted in accordance with OECD test guideline 203 in a GLP certified laboratory.

Based on the results of the range finding test, the definitive test was carried out using semi-static method at the test concentration of 100 mg/L along with a control. Seven fish were used for each test concentration.

There were no mortality and toxic signs observed in control group as well as test item treated group of 100 mg/L up to 96 h exposure. The test medium was found to be solution (red in colour) with the absence of settable solids at the test concentration of 100 mg/L.

In conclusion, the NOEC (No Observed Effect Concentration) and LC50 for FAT 40875/A TE at 96 hours was higher than 100 mg test item/L based on nominal concentration.