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 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:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation 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 as per Certificate of Analysis (Content): 86.4% all organic components
Batch No: BOP 05-17 (BS-ROE 1550 NIM 25+26+27)
Manufactured Date: December 20, 2017
Expiry Date: December 11, 2022
Recommended Storage Condition : Refrigeration (+2 to +8°C)
pH: 4.3 of 2% (w/w) aq. Solution.
Solubility: 499.9 g/L

Analytical monitoring:

yes

Details on sampling:

The prepared test concentration along with the negative control was analyzed for the test item concentration at the beginning and end of test. For analysis, single composite sample was drawn from prepared test concentration and negative control.

Vehicle:

yes

Remarks:

reconstituted water

Details on test solutions:

To prepare test concentrations of 6.25, 12.5, 25, 50 and 100 mg/L, 6.25, 12.5, 25, 50 and 100 mg of test item was added and made up to 1 L in a volumetric flask using test medium separately.

For negative control (G1), 1 L test medium was used.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Age at study initiation (mean and range, SD): Less than 24 hours (Neonates)
- Source: Ecotoxicology lab, Eurofins Advinus Ltd, Bengaluru, India.
- Feeding during test: Chlorella vulgaris.
- Food type: Unicellular green alga.

ACCLIMATION
- Acclimation conditions (same as test or not): Yes
- Health during acclimation (any mortality observed): No

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

Total hardness as Calcium carbonate: 2.35 m moles/L

Test temperature:

Range finding : 20.3 to 20.5 °C
Definitive test : 21.1 to 22.2 °C

pH:

Range finding : 7.69 to 7.81
Definitive test : 7.71 to 7.82

Dissolved oxygen:

Range finding : 7.0 to 7.4
Definitive test : 7.0 to 7.3

Conductivity:

The conductivity of the de-ionized water used for the preparation of test medium was <5 μS/cm.

Nominal and measured concentrations:

Definitive test: 0 (negative control), 6.25, 12.5, 25, 50 and 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: beakers
- Material, size, headspace, fill volume: 250mL, glass,
- Volume of solution: Fill volume - 15 mL in range finding test and 30 mL in definitive test.
- Aeration: No
- No. of organisms per vessel:
Range finding test: Ten daphnia per group divided into two replicates of five daphnia each.
Definitive test: Twenty daphnia per group divided into four replicates of five daphnia each.
- No. of vessels per control / vehicle control (replicates): 150 mL of prepared negative control, test item concentrations and reference substance was dispensed separately into 4 test containers.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Using water deionizing unit - Milli-Q.
- Alkalinity: 0.78 m moles/L
- Ca/mg ratio: 5.3 : 1
- Conductivity: The conductivity of the de-ionized water used for the preparation of test medium was <5 μS/cm.

OTHER TEST CONDITIONS
- Light intensity: complete darkness

VEHICLE CONTROL PERFORMED: yes

RANGE-FINDING STUDY
- Test concentrations: 0.00001, 0.0001, 0.001, 0.01, 0.1, 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: No immobility of the daphnia was observed in the negative control. The
observed percent immobilization of daphnia at 48 hour was 10 % at 100 mg/L, no immobilization was observed at any other concentrations at 24 and 48 hours of exposure.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Range finding study : No immobility of daphnia was observed in negative control and the tested concentrations.

Results with reference substance (positive control):

At 0.32 mg/L, the immobilization of daphnia was 40 and 50 % at 24 and 48 hours exposure, respectively.

Reported statistics and error estimates:

Since, the observed immobilization of daphnia was less than 50% in the highest test concentration used in the study no statistical analysis was
applied.

Definitive Test – Results on Daphnia Immobilization

Group	Treatment (mg/L)	No of Daphnia immobilized (5 daphnia per replicate)								% Immobilization
		24 h				48 h				24 h	48 h
		R1	R2	R3	R4	R1	R2	R3	R4
G1	Negative control	0	0	0	0	0	0	0	0	0	0
G2	6.25	0	0	0	0	0	0	0	0	0	0
G3	12.5	0	0	0	0	0	0	0	0	0	0
G4	25	0	0	0	0	0	0	0	0	0	0
G5	50	0	0	0	0	0	0	0	0	0	0
G6	100	1	1	0	1	1	1	2	1	15	25
G7	Reference Substance 0.32	2	3	2	1	3	3	2	2	40	50

Validity criteria fulfilled:

yes

Conclusions:

The EC50 value for FAT 40875/A TE at 48 hours was found to be greater than 100 mg/L.

Executive summary:

The acute immobilization effect of the test item FAT 40875/A TE was studied on Daphnia magna according to OECD test guideline 202, in a GLP certified laboratory.

A range finding test was carried out to determine the test concentrations for the definitive test, where Daphnia magna less than 24 hours old were exposed to the test item concentrations of 0.00001, 0.0001, 0.001, 0.01, 0.1, 1.0, 10 and 100 mg/L along with a negative control in test medium (reconstituted water) at 20.3 to 20.5°C.

The observed percent immobilization of daphnia at 48 hour was 0, 0, 0, 0, 0, 0, 0 and 10 % at 0.00001, 0.0001, 0.001, 0.01, 0.1, 1.0, 10 and 100 mg/L, respectively.

Based on the results of range finding test, in definitive test, Daphnia magna less than 24 hours old were exposed to test concentrations of 6.25, 12.5, 25, 50 and 100 mg/L along with a negative control and a reference substance, potassium dichromate at 0.32 mg/L in test medium (reconstituted water) at 21.1 to 22.2 °C. The number of daphnia immobilized was recorded at 24 and 48 hours exposure.

There was no immobilization of daphnia in the negative control. The observed per cent immobilization of daphnia was 0, 0, 0, 0 and 15 % on at 24h and 0, 0, 0, 0 and 25 % at 48 h exposure at the test concentration of 6.25, 12.5, 25, 50 and 100 mg/L, respectively.

At 0.32 mg/L reference substance, the immobilization of daphnia was 40 and 50 % at 24 and 48 hours exposure, respectively.

In conclusion, the EC50 value for FAT 40875/A TE at 48 hours was established to be greater than 100 mg/L.

Description of key information

The EC50 value for FAT 40875/A TE at 48 hours was established to be greater than 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The acute immobilization effect of the test item FAT 40875/A TE was studied onDaphnia magnaaccording to OECD test guideline 202, in a GLP certified laboratory.

A range finding test was carried out to determine the test concentrations for the definitive test, whereDaphnia magnaless than 24 hours old were exposed to the test item concentrations of 0.00001, 0.0001, 0.001, 0.01, 0.1, 1.0, 10 and 100 mg/L along with a negative control in test medium (reconstituted water) at 20.3 to 20.5°C.

The observed percent immobilization ofdaphniaat 48 hour was 0, 0, 0, 0, 0, 0, 0 and 10 % at 0.00001, 0.0001, 0.001, 0.01, 0.1, 1.0, 10 and 100 mg/L, respectively.

Based on the results of range finding test, in definitive test,Daphnia magnaless than 24 hours old were exposed to test concentrations of 6.25, 12.5, 25, 50 and 100 mg/L along with a negative control and a reference substance, potassium dichromate at 0.32 mg/L in test medium (reconstituted water) at 21.1 to 22.2 °C. The number of daphnia immobilized was recorded at 24 and 48 hours exposure.

There was no immobilization of daphnia in the negative control. The observed per cent immobilization of daphnia was 0, 0, 0, 0 and 15 % on at 24h and 0, 0, 0, 0 and 25 % at 48 h exposure at the test concentration of 6.25, 12.5, 25, 50 and 100 mg/L, respectively.

At 0.32 mg/L reference substance, the immobilization of daphnia was 40 and 50 % at 24 and 48 hours exposure, respectively.

In conclusion, the EC50 value for FAT 40875/A TE at 48 hours was established to be greater than 100 mg/L.