AVITERA ROSE SE

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:

Study initiation date - 05 March 2021; Experiment start date - 16 April 2021; Experiment completion date - 07 July 2021; Study completion date - 30 July 2021.

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:

yes

Remarks:

Due to unavoidable circumstances, experiment is delayed Experimental Completion Date June 07, 2021

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification: FAT 40879/B TE
Appearance: Dark red powder
Batch No.: BOP 05-20 (MC-0065114700/800 1:1)
Purity: 80.5 % all colored organic constituents; main constituent 66.9 %
Manufacture Date: 26 June 2020.
Expiry Date: 08 September 2025
Storage Conditions: Freezer (-15 to -20 °C)

Analytical monitoring:

yes

Details on sampling:

Test item was checked for its stability in dilution water (M4 medium) at 0, 24 and 48 hour.

Vehicle:

yes

Remarks:

M4 Medium

Details on test solutions:

Main Experiment : 1, 2.20, 4.84, 10.65, 23.43 and 51.54 mg/L .

Test organisms (species):

Daphnia magna

Details on test organisms:

Test system (Species): Daphnia magna
Age: Young Daphnids, aged less than 24 hours old.
Justification: Recommended by the guideline.
Source: In - house culture, RCC Laboratories India Private Limited (Originally procured from Department of Advanced Zoology & Biotechnology, Ethiraj College for Women, Chennai)

Test type:

semi-static

Water media type:

freshwater

Remarks:

M4 Medium

Limit test:

no

Total exposure duration:

48 h

Hardness:

Main Experiment: 210 - 220 mg/L

Test temperature:

Main Experiment:
Control: 20.7 - 20.9 °C,
Treatment: 20.7 - 20.9 °C

pH:

Main Experiment: Control: 7.14 - 7.25; Treatment: 6.97 - 7.28

Dissolved oxygen:

Main Experiment: Control: 7.68 - 7.80 mg/L; Treatment: 7.54 - 7.67 mg/L

Nominal and measured concentrations:

1.0, 2.20, 4.84, 10.65, 23.43 and 51.54 mg/L. (Nominal concentrations)
0.85, 1.84 ,4.25, 9.99, 16.61 and 31.84 mg/L (Time weighted mean Concentrations).

Details on test conditions:

TEST SYSTEM
- Test vessel: Beakers
- Type: 100mL Glass beakers
- Material, size, headspace, fill volume: 100mL
- Volume of solution: 50mL
- Aeration: No
- No. of organisms per vessel: 10 Daphnids.
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: M4 medium

OTHER TEST CONDITIONS
- Photoperiod: 16 hour light and 8 hour darkness

EFFECT PARAMETERS MEASURED:
VEHICLE CONTROL PERFORMED: Yes

RANGE-FINDING STUDY
- Test concentrations: 0.1, 1, 10, 50 and 100 mg/L.
- Results used to determine the conditions for the definitive study: Immobilisation at 24 and 48 hours.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

>= 7.41 - < 8.71 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Remarks:

Time weighted mean Concentrations 0.85, 1.84, 4.25, 9.99, 16.61 & 31.84 mg/L.

Basis for effect:

mobility

Remarks:

Immobilisation

Details on results:

MAIN EXPERIMENT
No immobilisation was observed in control and daphnids exposed to control, 0.85 and 1.84 mg/L time weighted mean concentrations throughout the experimental period. At the end of 24-hour testing period, 2/40, 3/40, 12/40 and 20/40 immobility were observed in daphnids exposed to 4.25, 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations, respectively. At the end of 48-hour testing period, 10/40, 28/40, 33/40 and 40/40 cumulative immobility was recorded in the daphnids exposed to 4.25, 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations respectively. Therefore, the percent cumulative immobilisation at the end of 48 hours was recorded to be 0 % in control and daphnids exposed to 0.85 and 1.84 mg/L time weighted mean concentrations whereas, 25 %, 70 %, 82.5 % and 100 % was observed in daphnids exposed to 4.25, 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations, respectively.

- Behavioural abnormalities: Lethargy
- Other biological observations: No
- Mortality of control: No
- Other adverse effects control: No
- Immobilisation of control: No
- Abnormal responses: No
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Possibly Environmental parameters like temperature.
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

An acute immobilisation toxicity study (Study No. 10811) in Daphnia magna was performed with Potassium dichromate (Batch No. DK9D692695) in accordance with the guideline OECD 202. Daphnids were exposed to a range of five different concentrations such as 0.4, 0.72, 1.30, 2.33 and 4.20 mg/L. Four replicates (5 daphnids in each replicate) were maintained for each test concentration, concurrently control group was maintained along with the concentrations. The test item was formulated with M4 medium. No immobilisation was observed in control and daphnids exposed to 0.4 mg/L throughout the experimental period. At the end of 24 hour, 2/20, 6/20, 12/20 and 20/20 immobility was observed in daphnids exposed to 0.72, 1.30, 2.33 and 4.20 mg/L concentrations, respectively. Thus, the percent immobilisation at the end of 24 hour was recorded to be 0 % in control and daphnids exposed to 0.4 mg/L concentration whereas, 10 %, 30 %, 60 % and 100 % immobility was observed in daphnids exposed to 0.72, 1.30, 2.33 and 4.20 mg/L concentrations, respectively. At the end of 48 hours, 3/20, 10/20 and 17/20 cumulative immobility was observed in daphnids exposed to 0.72, 1.30 and 2.33 mg/L concentrations, respectively. Thus, the percent immobilisation at the end of ‘48’ hour was observed to be 0 % in control and daphnids exposed to 0.4 mg/L concentration whereas, 15 %, 50 %, 85 % and 100 % was observed in daphnids exposed to 0.72, 1.30, 2.33 and 4.20 mg/L concentrations, respectively. No toxicity signs were observed in control and daphnids exposed to 0.4 and 0.72 mg/L concentrations throughout the experiment. At the end of 24 and 48 hour, lethargy was observed in daphnids exposed to 1.30 and 2.33 mg/L concentrations.

- Results with reference substance valid? Yes
- Limit test: No

Reported statistics and error estimates:

At ‘48’ hour, the EC50 was calculated as 1.29 mg/L with upper confidence limits of 1.57 mg/L and lower confidence limits of 1.07 mg/L and regression equation Y = 4.4835 + 4.5697 log (x). Based on immobility, NOEC was determined to be 0.4 mg/L and LOEC was determined to be 0.72 mg/L.

Validity criteria fulfilled:

yes

Remarks:

In control, no immobilization was observed throughout the experimental period.

Conclusions:

The EC50 of FAT 40879/B in Daphnia magna observed for a period of 48 hour is calculated as 7.41 mg/L with upper confidence limits of 8.71 mg/L and lower confidence limits of 6.24 mg/L and regression equation Y = 2.0111 + 3.4362 log (x). Based on the immobility at 48 hours, NOEC is 1.84 mg/L and LOEC is 4.25 mg/L time weighted mean concentrations, respectively.

Executive summary:

In a GLP-study, Acute Immobilisation Toxicity Study in Daphnia magna was conducted in accordance with OECD guideline 202 with FAT 40879/B. Test item was found to be soluble in M4 medium. Stability of the test item in the M4 medium was performed with a nominal concentration of 100.03 mg/L at 0 hour, 24 hour and 48 hour and it was stable up to 24 hours in the M4 Medium. Hence, semi-static test with 24 hours medium renewal was performed. Initially, a range finding experiment was conducted with a range of concentrations such as 0.1, 1, 10, 50 and 100 mg/L. No immobilisation was observed in control and daphnids exposed to 0.1 and 1 mg/L test concentrations and 100 % immobilisation was observed in daphnids exposed to 50 and 100 mg/L test concentrations. Therefore, based on the results of range finding experiment, main experiment was conducted with six different test concentrations such as 1, 2.20, 4.84, 10.65, 23.43 and 51.54 mg/L. The daphnids with brood chambers were acclimatized one day prior to the exposure in M4 medium. The test item was formulated with M4 Medium. Forty daphnids with four replicates (10 daphnids in each replicate) for each test concentration and control group were maintained. After the exposure on day 0, daphnids were observed for immobilisation approximately at 24 and 48 hour and the respective data were recorded. Water Parameters such as pH, temperature and dissolved oxygen were recorded at the start and during test medium renewal (both old and fresh media) and at completion in control and all test concentrations. Hardness was analyzed in the fresh media of the control. Test solution was renewed at 24 hour and most of the measured concentrations of old samples were deviated more than 20 % from the nominal concentration of fresh samples. Hence, time weighted mean concentration is calculated and represented for the interpretation of the results of main experiment.

 

Immobilisation: 

No immobilisation was observed in control and daphnids exposed to control, 0.85 and 1.84 mg/L time weighted mean concentrations throughout the experimental period. At the end of the 24-hour incubation period, 2/40, 3/40, 12/40 and 20/40 immobility were observed in daphnids exposed to 4.25, 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations respectively. At the end of the 48-hour incubation period, 10/40, 28/40, 33/40 and 40/40 cumulative immobility was recorded in the daphnids exposed to 4.25, 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations respectively. Therefore, the percent cumulative immobilisation at the end of 48 hours was recorded to be 0 % in control and daphnids exposed to 0.85 and 1.84 mg/L time weighted mean concentrations whereas, 25 %, 70 %, 82.5 % and 100 % was observed in daphnids exposed to 4.25, 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations, respectively.

 

Toxicity Signs:

No toxicity signs were observed in control and daphnids exposed to 0.85 and 1.84 mg/L time weighted mean concentrations throughout the experimental period. At the end of 24 hours, lethargy was observed in daphnids exposed to 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations. At the end of 48 hours, lethargy was observed in daphnids exposed to 9.99 and 16.61 mg/L time weighted mean concentrations.

 

Conclusion: 

The EC₅₀ of FAT 40879/B in Daphnia magna observed for a period of 24 and 48 hour was calculated based on the time weighted mean concentration. The EC₅₀ of FAT 40879/B in Daphnia magna observed for a period of 24 hour is 31.88 mg/L with upper confidence limits of 51.74 mg/L and lower confidence limits of 24.04 mg/L and regression equation Y = 1.6662 + 2.2173 log (x). 

The EC₅₀ of FAT 40879/B in Daphnia magna observed for a period of 48 hour is calculated as 7.41 mg/L with upper confidence limits of 8.71 mg/L and lower confidence limits of 6.24 mg/L and regression equation Y = 2.0111 + 3.4362 log (x).

Description of key information

The EC₅₀ of FAT 40879/B in Daphnia magna observed for a period of 48 hour is calculated as 7.41 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

7.41 mg/L

Additional information

In a GLP-study, Acute Immobilisation Toxicity Study in Daphnia magna was conducted in accordance with OECD guideline 202 with FAT 40879/B. Test item was found to be soluble in M4 medium. Stability of the test item in the M4 medium was performed with a nominal concentration of 100.03 mg/L at 0 hour, 24 hour and 48 hour and it was stable upto 24 hours in the M4 Medium. Hence, semi-static test with 24 hours medium renewal was performed. Initially, a range finding experiment was conducted with a range of concentrations such as 0.1, 1, 10, 50 and 100 mg/L. No immobilisation was observed in control and daphnids exposed to 0.1 and 1 mg/L test concentrations and 100 % immobilisation was observed in daphnids exposed to 50 and 100 mg/L test concentrations. Therefore, based on the results of range finding experiment, main experiment was conducted with six different test concentrations such as 1, 2.20, 4.84, 10.65, 23.43 and 51.54 mg/L. The daphnids with brood chambers were acclimatized one day prior to the exposure in M4 medium. The test item was formulated with M4 Medium. Forty daphnids with four replicates (10 daphnids in each replicate) for each test concentration and control group were maintained. After the exposure on day 0, daphnids were observed for immobilisation approximately at 24 and 48 hour and the respective data were recorded. Water Parameters such as pH, temperature and dissolved oxygen were recorded at the start and during test medium renewal (both old and fresh media) and at completion in control and all test concentrations. Hardness was analyzed in the fresh media of the control. Test solution was renewed at 24 hour and most of the measured concentrations of old samples were deviated more than 20 % from the nominal concentration of fresh samples. Hence, time weighted mean concentration is calculated and represented for the interpretation of the results of main experiment.

 

Immobilisation: 

No immobilisation was observed in control and daphnids exposed to control, 0.85 and 1.84 mg/L time weighted mean concentrations throughout the experimental period. At the end of 24 hour, 2/40, 3/40, 12/40 and 20/40 immobility were observed in daphnids exposed to 4.25, 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations respectively. At the end of 48 hour, 10/40, 28/40, 33/40 and 40/40 cumulative immobility was recorded in the daphnids exposed to 4.25, 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations, respectively. Therefore, the percent cumulative immobilisation at the end of 48 hours was recorded to be 0 % in control and daphnids exposed to 0.85 and 1.84 mg/L time weighted mean concentrations whereas, 25 %, 70 %, 82.5 % and 100 % was observed in daphnids exposed to 4.25, 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations, respectively.

 

Toxicity Signs:

No toxicity signs were observed in control and daphnids exposed to 0.85 and 1.84 mg/L time weighted mean concentrations throughout the experimental period. At the end of 24 hours, lethargy was observed in daphnids exposed to 9.99, 16.61 and 31.84 mg/L time weighted mean concentrations. At the end of 48 hours, lethargy was observed in daphnids exposed to 9.99 and 16.61 mg/L time weighted mean concentrations.

 

Conclusion: 

The EC₅₀ of FAT 40879/B in Daphnia magna observed for a period of 24 and 48 hours was calculated based on the time weighted mean concentration. The EC₅₀ of FAT 40879/B in Daphnia magna observed for a period of 24 hours is 31.88 mg/L with upper confidence limits of 51.74 mg/L and lower confidence limits of 24.04 mg/L and regression equation Y = 1.6662 + 2.2173 log (x). 

The EC₅₀ of FAT 40879/B in Daphnia magna observed for a period of 48 hours is calculated as 7.41 mg/L with upper confidence limits of 8.71 mg/L and lower confidence limits of 6.24 mg/L and regression equation Y = 2.0111 + 3.4362 log (x).