Phosphonium, tetraphenyl-, salt with 4,4'-sulfonylbis[phenol], compd. with 4,4'-sulfonylbis[phenol] (2:2:1)

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:

15 Feb 2022 to 18 Feb 2022

Reliability:

1 (reliable without restriction)

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:

Lot No.: 108404E
Appearance: White powder
Expiry date: August 03, 2022
Storage: Room temperature, protected from light

Analytical monitoring:

yes

Details on sampling:

SAMPLING
For determination of the test item concentrations in the test solutions, all four test vessels were
analysed at the start and at the end of the test. In case of the control group two samples were
analysed at the start and at the end of the test.

Vehicle:

no

Details on test solutions:

The test item was completely and evenly suspended in ISO medium by mechanical dispersion without the use of any solubilising agent in the testing laboratory (Validation of the Analytical Method for the Determination of SBCAT-03 for the Eco-toxicological Studies performed by TOXI-COOP ZRT., Study No.: 950-100-6322).
0.0400 g of test item was suspended in 800 mL of the dilution water (ISO Medium; see section 5.4) in order to give the nominal loading rate of 50 mg/L of the test item and handled in ultrasonic bath for approximately 60 minutes. Thereafter, this stock solution was filtrated through a membrane filter (0.45 µm)* to separate the possible non-dissolved test material. The further test item concentrations were prepared from this test item stock solution (TISS) as shown in any other information on materials
*Type: Thermo NALGENE® Lot number: 1298115
Expiration date: September 2025

Test organisms (species):

Daphnia magna

Details on test organisms:

Species and strain: Daphnia magna (Straus)
Origin: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection)
2100 Gödöllő, Kotlán S. u. 3. Hungary
Breeding: The Daphnia are bred in the Laboratory of TOXI-COOP ZRT.
Number of animals: Twenty animals in every groups (test concentrations and untreated control), divided into 4 replicates (5 animals per replicates).
Age of animals: Less than 24 h old at the start of exposure period. Sex: Female
Animal health: Apparently healthy animals were used in this test with a known history Acclimatization: Test animals were bred under similar (or the same) conditions as that used during the exposure period (holding water, temperature,
background colour etc.), therefore additional acclimatisation before the test was not necessary.
Food and Feeding: The Daphnia were fed with concentrated algal suspension of Raphidocelis subcapitata during the holding. Test animals were not fed during the exposure.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

The measured hardness of the ISO Medium used in the test was determined to be
246.174 mg/L (as CaCO3).

Test temperature:

The test temperature was in the range of 19.8 – 20.9 °C measured in the test vessels during the test. The additionally measured ambient temperature within the climate chamber was between 19.8 – 20.8 °C.

pH:

The pH of the test solutions was not adjusted and did not vary by more than 1.5 units in any one test. The pH was in the range of 7.68 - 8.12 during the test.

Dissolved oxygen:

The dissolved oxygen concentration was in the range of 8.01 –8.28 mg/L during the test.

Nominal and measured concentrations:

See any other information on materials

Details on test conditions:

The test vessels were kept in a climate chamber under controlled environmental conditions during the test and test solutions were not aerated. The water temperature, dissolved oxygen concentration and pH were measured in each test group at the start (before test solutions had been distributed into the test vessels) and in each test vessel at the end of the experiment. Additionally, the ambient temperature was measured continuously using a min/max thermometer in the climate chamber.
Test type: Static test. Based on the results obtained during analytical method validation (Study number: 950-100-6322) the test item is stable for the duration of 48 hours in ISO Medium.
Temperature: The test temperature was in the range of 19.8 – 20.9 °C measured in the test vessels during the test. The additionally measured ambient temperature within the climate chamber was between 19.8 – 20.8 °C.
Oxygen Concentration: The dissolved oxygen concentration was in the range of 8.01 –
8.28 mg/L during the test.
pH: The pH of the test solutions was not adjusted and did not vary by more than 1.5 units in any one test. The pH was in the range of 7.68 - 8.12 during the test.
The data of pH, dissolved oxygen concentration and water temperature measured during the test are detailed in Appendix I; Table 6.
Light-Dark Cycle: The light-dark cycle during the test was 16 hours light (artificial illumination) and 8 hours darkness. The light intensity measured at the position occupied by the test beakers was 516 lux at the start of the test.
Hardness: The measured hardness of the ISO Medium used in the test was determined to be
246.174 mg/L (as CaCO3).

Reference substance (positive control):

yes

Remarks:

For the evaluation of the quality of the Daphnia clone and the experimental conditions, Potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1.563 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

3.125 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

2.268 mg/L

95% CI:

> 1.536 - < 2.781

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

3.708 mg/L

95% CI:

> 3.085 - < 4.451

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Immobilisation was 5 % (< 10 %) in the control group and none of the daphnids showed signs of disease or stress. The dissolved oxygen concentration during the test in control and test vessels was in the range of 8.01 – 8.28 mg/L (more than 3 mg/L in all cases). All validity criteria were within acceptable limits and therefore the study is considered as valid.

10.2 Analytical Results

For determination of the test item concentrations in the test solutions, all four test vessels were analysed at the start and at the end of the test. In case of the control group two samples were analysed at the start and at the end of the test.
Signal intensities measured for the untreated control samples were below the quantification limit (i.e. no interfering component was detected in the untreated control samples).
The mean of the measured concentrations of the test item were in the range of 93 – 98 % of the nominal concentrations at the start and between 98 – 105 % at the end of the experiment. Therefore, the biological results are based on the nominal test item concentrations.
Detailed description of the analytical part of the study (method, determination, results) is attached to the Report as Appendix III.

10.3 Biological Results

After 48 hours exposure one immobile animal was observed in the control group and in the lowest test item group of 1.563 mg/L (5 % immobilisation). 40 % immobilisation was detected at the test concentration of 3.125 mg/L, while at 6.25 mg/L the immobilisation was 90 %. In the test groups of 12.5, 25 and 50 mg/L 100 % of the test animals was immobilized
All biological results are based on the nominal test item concentrations.

Results with reference substance (positive control):

The 24h EC50 value was determined to be 0.92 mg/L (95% conf. limits: not determined).

Reported statistics and error estimates:

The 48-h ECx values and their 95 %-confidence limits were calculated by Probit analysis (statistical software program ToxRatPro Version 3.3.0). NOEC and LOEC values were determined on the basis of Step-down Cochran-Armitage Test (α = 0.05; statistical software program ToxRatPro Version 3.3.0). See "illustration" for details.

Summary of the Biological Endpoints

Endpoint	Concentration (mg/ml) based on nominal concentrations
48 h-EC10 95 % confidence limits	2.268 1.536 – 2.781
48 h-EC20 95 % confidence limits	2.685 1.988 – 3.208
48 h-EC50 95 % confidence limits	3.708 3.085 - 4.451
48 h-NOEC	1.563
48 h-LOEC	3.125

Validity criteria fulfilled:

yes

Conclusions:

All validity criteria were met and therefore the study is considered as valid. Biological endpoints of the study are summarised in any other information on results
In this 48-hour acute toxicity test on Daphnia magna, the obtained results showed that the test item SBCAT-03 had significant toxic effects on the mobility of Daphnia.
The 48-h EC50 value was determined to be 3.708 mg/L. The 48-h NOEC was determined to be 1.563 mg/L. The 48-h LOEC was determined to be 3.125 mg/L.
All validity criteria were met. The results are based on nominal test item concentrations.

Executive summary:

Acute toxicity of the test item SBCAT-03 was assessed on Daphnia magna in a 48-hour acute immobilisation test. Young Daphnia were exposed to aqueous test media containing the test item for 48 hours in a static test, as the analytical method validation (950-100-6322) demonstrates a sufficient stability of the test item in the test medium.

The test animals were exposed to aqueous test media containing the test item for 48 hours at a range of concentrations (1.563, 3.125, 6.25, 12.5, 25 and 50 mg/L) plus a control. The concentration range was chosen based on results obtained in a non-GLP pre-test.

 

The quantification of the test item SBCAT-03 was performed by a previously validated analytical method by the analytical laboratory of TOXI-COOP ZRT. Samples were taken from the test concentrations and from the control at the start and at the end of the experiment and analysed by HPLC method. Mean of the measured test item concentrations remained within

± 20 % of the nominal over the test period of 48 hours, therefore the biological results are based on the nominal test concentrations.

 

Healthy, young female daphnids < 24 hours of age and with known origin, history and acclimatisation conditions were used. Twenty Daphnia (divided into 4 replicates) were tested each, exposed to the test concentrations and to the control medium. Each test vessel contained approximately 40 mL test medium. Fully characterized (content, physico-chemical characteristics) ISO Medium was used as test medium.

 

The immobilisation of the test animals was observed 24 and 48 hours after test start. Environmental conditions were recorded at the start and at the end of the experiment. All measured values remained within the acceptable ranges.

 

After 48 hours exposure one immobile animal was observed in the control group and in the lowest test item group of 1.563 mg/L (5 % immobilisation). 40 % immobilisation was noted at the test concentration of 3.125 mg/L, while at 6.25 mg/L the immobilisation was 90 %. In the test groups of 12.5, 25 and 50 mg/L 100 % of the test animals was immobilized. Accordingly, the 48h NOEC was determined to be 1.563 mg/L, while the 48-h LOEC was determined to be 3.125 mg test item/L. The 48-h-EC50 was calculated to be 3.708 mg/L.

 

The 48-h ECx values were calculated by Probit analysis (statistical software program ToxRatPro Version 3.3.0). NOEC and LOEC values were determined on the basis of Step- down Cochran-Armitage Test (α = 0.05; statistical software program ToxRatPro Version 3.3.0).

All validity criteria were met and therefore the study is considered as valid (see section 10.1) The biological endpoints are summarized below

Endpoint	Concentration (mg/ml) based on nominal concentrations
48 h-EC10 95 % confidence limits	2.268 1.536 – 2.781
48 h-EC20 95 % confidence limits	2.685 1.988 – 3.208
48 h-EC50 95 % confidence limits	3.708 3.085 - 4.451
48 h-NOEC	1.563
48 h-LOEC	3.125

Description of key information

In this 48-hour acute toxicity test on Daphnia magna, the obtained results showed that
the test item SBCAT-03 had significant toxic effects on the mobility of Daphnia.
The 48-h EC50 value was determined to be 3.708 mg/L.
The 48-h NOEC was determined to be 1.563 mg/L.
The 48-h LOEC was determined to be 3.125 mg/L.
All validity criteria were met. The results are based on nominal test item concentrations.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

3.708 mg/L