2-(1-(2-hydroxy-3,5-di-tert-pentyl-phenyl)ethyl)-4,6-di-tert-pentylphenyl acrylate

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26.02.2010-16.04.2010

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Remarks:

certificate included in the full study report

Test material

Test material information

Constituent 1

Reference substance name:

2-(1-(2-hydroxy-3,5-di-tert-pentyl-phenyl)ethyl)-4,6-di-tert-pentylphenyl acrylate

EC Number:

413-850-6

EC Name:

2-(1-(2-hydroxy-3,5-di-tert-pentyl-phenyl)ethyl)-4,6-di-tert-pentylphenyl acrylate

Cas Number:

123968-25-2

Molecular formula:

C37H56O3

IUPAC Name:

2-{1-[2-hydroxy-3,5-bis(2-methylbutan-2-yl)phenyl]ethyl}-4,6-bis(2-methylbutan-2-yl)phenyl prop-2-enoate

Constituent 2

Reference substance name:

Zikanox-549

IUPAC Name:

Zikanox-549

Details on test material:

Name: ZIKANOX-549
Manufacturer & Supplier: ZIKO Ltd.
Manufacture date: 2009-09-10
Receipt date: 2009-10-27
Receipt quantity: 248.988 g (gross weight)
Appearance: off white powder
Lot Number: 10909001-003
Purity: 99.5 %
Storage condition: Room temperature
Expiration date (after manufacture): 3 years
- The test substance was prepared by dispersing with acetone and M4 medium and sonicating for 30 minutes as the test substance was insoluble in water. The test substance was prepared without adjustment of pH.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The analyses of test substance in the test medium were performed at the test start (0 hour) and the test end (48 hours)

Test solutions

Vehicle:

yes

Details on test solutions:

- Preparation of test solution: 0.101 g of test substance was put into 1000 mL of volumetric flask and added 0.100 g of Acetone. Then test water was filled up to the 1000 mL marker to prepare stock solutions (stock solution, 100 mg/L). The stock solution was dispersed with ultrasonicator (1510E -DTH, Branson ultrasonic Co.) for 30 minutes and filtered through 0.45 μm membrane filter. The stock solution was divided 100 mL each to prepare test solution.
- Test concentration: According to the results of range-finding test ; 0.10, 1.00, 10.00 and 100.00 mg/L treatment, no immobilized daphnia was observed in the highest concentration, 100.0 mg/L. Therefore, the test was performed in 100.00 mg/L (nominal concentration) of Zikanox-549 as limit test.
- Control: Negative Control: M4 medium was used for control
- Solvent control: 0.100 g of acetone was put into 1000 mL of volumetric flask and test water was filled up to the 10 000 mL marker to prepare stock solution. The stock solution was divided 100 mL each to prepare solvent. The concentration of solvent was 100 mg/L in test water.
- Positive control: The test was performed in control, 0.20, 0.40, 0.80, 1.60 and 3.20 mg/L (geometric series 2.0, nominal concentration) as active ingredient of Potassium dichromate (Sigma-aldrich, Lot No. 09514PA) as positive (October 20, 2009 ~ October 22, 2009).

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

- Selection of the test species: The Daphnia magna are recommended species by OECD guideline for the testing of chemicals 202, "Daphnia sp., Acute Immobilisation Test" (Adopted: 13 April 2004) and "U.S. EPA Toxic Substances Control Act Test Guideline" (1985-09-27). And it was selected for this test because of a plenty of comparing toxicological data.
- Breeding condition: The daphnia were bred in 1 L water bath. The temperaure of breeding water was maintained between (19 ~ 21) °C and photo periods were 16 hours light and 8 hours dark. Feeds were supplied a time per day ; Chlorella vulgaris (ATCC, April 28, 2005) in the morning.
- Monitoring of breeding condition: The temperature of daphnia breeding room and breeding water were recorded automatically every 30 minutes. Photo periods were recorded every month. As the results of the monitoring, there was not observed an alteration affecting the test.
- Breeding water: The breeding water was prepared by OECD guidelines for the testing of chemicals 202, "Daphnia sp., Acute immobilisation test" Annex 3 Elendt M7 and M4 medium ( Adopted: 13 April 2004). The water quality was analyzed - water hardness by Inductively coupled plasma emission spectroscopy of Korea Standard M 0111 and alkalinity was assesed by titration method of Standard method (21th edition, APHA, AWWA, WEF, USA). As the results of the analysis, there was not observed the alteration affecting the test.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

no data

Test conditions

Hardness:

The total hardness of the test water: 187.3 mg/L CaCO3
The alkalinity: 55.1 mg/L

Test temperature:

Average temperature: 20.0 ±0.2 °C (19.8 °C - 20.6 °C)

pH:

Average pH: 8.13 (7.87 - 8.29)

Dissolved oxygen:

Average Dissolved Oxygen (DO): 7.91 mg/L (7.54 - 8.26 mg/L)
Percent of air saturation: 89.5 % (86.7 % ~ 92.6 %)

Salinity:

no data

Nominal and measured concentrations:

The toxicity test was performed in control and 100.00 mg/L (nominal concentration) of Zikanox-549 as limit test. And measured concentration is 0.008 mg/L

Details on test conditions:

Test is performed according to the OECD TG 202. Exposure condition: 20 daphnia less than 24 hours were divided into 4 groups of five animals each. 100 mL of M4 medium was filled up to the 135 mL cylindric glass bath (65 mm H. x 57 mm φ).

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.008 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Concentration of test substance in test solution: As the test result of analysis at 0 hours, the concentration of ZIKANOX-549 in test water was 0.009 mg/L and 0.0 % of nominal concentration. The concentration at 48 hours was 0.008 mg/L during the test periods.

- Immobility and abnormal behaviors: At the range-finding test, there was no immobility in any of daphnia exposed to (0.10 ~ 100.00) mg/L of test solution at 48 hours. The limit test was conducted in measured concentrations of 0.008 mg/L. After 48 hour exposure, no daphnia was immobilised in 0.008 mg/L of test solution. No abnormal behavior was observed in any of the test daphnia exposed to control, solvent control and treatment group during the test.

- Changes of water quality: During the definitive test period, the mean of pH was 8.13 (7.87 ~ 8.29), the mean of DO was 7.91 mg/L (7.54 mg/L ~ 8.26 mg/L) and per cent of air saturation value was 89.5 % (86.7 % - 92.6 %).

The mean of temperature was (20.0 ± 0.2) °C (19.8 ~ 20.6 °C). The total hardness of the test water 187.3 mg/L as CaCO3, the alkalinity was 55.1 mg/L.

Results with reference substance (positive control):

Reference substance : Potassium dichromate
24h-EC50 : 0.875mg/L (0.731-1.040)
48h-EC50 : 0.657mg/L(0.558-0.773)

Applicant's summary and conclusion

Conclusions:

The 24 hours and 48 hours-EC50 value of Zikanox-549 to Daphnia magna was investigated as higher than 0.008 mg/L. The NOEC of test substance to Daphnia magna was 0.008 mg/L at 24 hours and 48 hours. Determinations of EC50 and NOEC were based on measured concentration. There was not observed an alteration affecting the test in this test.

Executive summary:

The daphnia sp. Acute immobilisation test of Zikanox-549 was studied using Daphnia magna for 48 hours under the static system. Observations were made on the number of immobilized daphnia and abnormal behavior of Daphnia magna.

- This study was conducted in accordance with the OECD guidelines for the testing of chemicals 202, "Daphnia sp. Acute immobilisation test "(Adopted: 13 April, 2004).

- The toxicity test was performed in control and 100.00 mg/L (nominal concentration) of ZIKANOX-549 as limit test. The analyses of test substance in the test medium were performed at the test start (0 hour) and the test end (48 hours)

- As analysis results of ZIKANOX-549 in test solution, the mean concentrations of test substance were 0.008 mg/L.

- As the results of stability test of ZIKANOX-549 in the test water, the test substance was stable in the water during 48 hours. Therefore, the test was conducted under static conditions.

The test results are as follow:

Observation time	EC50 (mg/L)	95% confidence limits (mg/L)	NOEC (mg/L)
24 h	>0.008	-	0.008
48 h	>0.008	-	0.008

- No abnormal behavior was observed in any of the test daphnia exposed to control, solvent control and treatment group during the test.

- The mean of pH was 8.13 (7.87 ~ 8.29)

- The mean of Dissolved Oxygen concentration (DO) was 7.91 mg/L (7.54 mg/L ~ 8.26 mg/L) and percent of air saturation value was 89.5 % (86.7 % - 92.6 %)

- The mean of temperature was (20.0 ± 0.2) °C (19.8 °C ~ 20.6 °C)

Following the test results, 48 hours-EC50 value of the test substance was investigated at greater than 0.008 mg/L as measured concentration.