Rabitle FP-100

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: 16th July 2004 Experimental start date: 27th July 2004 Experimental end date: 29th July 2004 Study completion date: 19th August 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: - Study generated according to generally valid and/or internationally accepted testing guidelines - Performed according to GLP - Test parameters based on specific testing guideline - No GLP certificate provided

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

OECD Guideline No. 202 "Effects ob Biotic Systems: Acute Immobilisation Test and Reproduction Test: Daphnia sp" (1984 ).

Deviations:

no

Principles of method if other than guideline:

Also undertaken following procedures in:

Daphnia sp., Acute Immobilisation Test stipulated in the ‘Testing Methods for New Chemical Substances’ (November 21, 2003; No. 1121002, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare; November 13, 2003, No. 2, Manufacturing Industries Bureau, Ministry of the Environment)

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
N/A

Analytical monitoring:

not specified

Details on sampling:

See report

Vehicle:

yes

Details on test solutions:

Identity and concentration of auxiliary solvent for dispersal: N,N-Dimethylfomamide (DMF).
A stock solution was prepared with correcting by the purity (99.3%). Desired amount of test item supplied by sponsor was weighed and dissolved in DMF to prepare a stock solution of 1000 mg/L. A required quantity of the stock solution was added to the dilution water stirring in a container for preparation. The prepared test solution was divided into each test vessel.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (Clone A)
- Strain: No details provided in report
- Source: Young daphnids produced by parents which were cultured in the Kurume Laboratory were used. Daphnids originally came from the University of Sheffield (Sheffield S10 2UQ, UK).
- Age at study initiation (mean and range, SD): Less than 24 hours
- Method of breeding: The parents to obtain young daphnia were bred in the same quality of water (dechlorinated tap water), water temperature (20±1°C), and photoperiod as used in the test. Daphnia which age and survival rate in the lot were respectively 21-days old and greater than 100% was used for the test. Chlorella vulgaris of 0.1 to 0.2 mgC/day per daphnia was fed to the parents once a day. A 48-hour acute immobilisation test of potassium dichromate (Regent chemical, Wako Pure Chemical Industries, Ltd.) with the test organisms was conducted to confirm the reproducibility of the test system. The 48-hour EC50 of the reference substance was 0.283 mg/L. This value was within the normal range of reference substance in Kurume Laboratory (mean ± 2 SD,: 0.110 to 0.352 mg/L) [mean ± SD,: 0.231 ± 0.061 mg/L (n=45)].
- Feeding during test: Test organisms were not fed during the exposure
- Aeration: No aeration was used for the test during the exposure

ACCLIMATION – No details provided in report

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

N/A

Hardness:

CA 12 mg CaCO3/L

Test temperature:

20±1°C (19.8 – 19.9°C)

pH:

7.4 to 7.7

Dissolved oxygen:

60% or above of the air saturation value at the water temperature used (8.6 – 8.9 mg/L). Measured DO values met the criterion for the validity of the test (ie. not less than 60% of air saturation vale at the water temperature – saturated DO concentration (19-21°C) = 9.01 – 8.68 mg/L (JIS K 0102))

Salinity:

N/A

Nominal and measured concentrations:

One exposure level (nominal concentration: 0.100 mg/L)

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL beaker
- Type (delete if not applicable): open / closed – Test vessels were covered with lids in order to prevent dust, and evaporation of the test solutions
- Renewal rate of test solution (frequency/flow rate): Test solutions were renewed 24 hours after preparation, as a semi-static method
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated tap water, with sufficient aeration and controlled temperature, was used. It was confirmed before use that the concentration of residual chlorine was less than 0.02 mg/L. See Appendix 1 of report for chemical characteristics of dilution water.
- Intervals of water quality measurement: DO, pH and water temperature of test solution were measured at the start of exposure (at the preparation), before (after 24h) and after the renewal (at the preparation), and at the end of the exposure.

OTHER TEST CONDITIONS
- Adjustment of pH: No adjustment of pH was applied for the test solution
- Photoperiod: 16-hour light/8-hour dark photoperiod daily with a room lamp

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): EC50 (24 and 48h) and NOEC (48h)

TEST CONCENTRATIONS
- The test concentration was decided based on results of preliminary tests and solubility test. The test concentrations were expressed as value corrected by the purity (99.3%) of the test item. See report for results of preliminary tests.

- Reason for setting test concentration: The solubility of test item to dilution water was not reproducible and it was not able to determine a constant solubility. The concentration in dispersed suspension containing test item was not detected. So the test solution was prepared using a solvent. The 0.100 mg/L nominal concentration which was expected to maintain the concentration in test solution prepared with N, N-Dimethylfomamide (DMF) during a period and be more than the solubility was used in the test.

Reference substance (positive control):

no

Remarks:

water

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 0.05 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.05 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.05 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Immobility – no abnormal responses were observed in the present test concentration during the exposure. Immobility in control was 0%, which meets the criterion for the validity of the test (ie. less than 10%), and no floating daphnids were observed.

Abnormal response – no abnormal response were obtained in the present test concentration during the exposure compared with the control. Also no abnormal response observed during the exposure in the control.

Results with reference substance (positive control):

N/A

Reported statistics and error estimates:

Values were rounded off in accordance with JIS Z 8401 rule B (JIS = Japanese Industrial Standards)

See Reprt

Validity criteria fulfilled:

yes

Executive summary:

A 48-hour acute Immobilization test of FP-100 with Daphnia magna was conducted.   Twenty test organisms were used in one exposure level (nominal concentration: 0.100 mg/L), a vehicle control and a control. The water temperature was maintained within the range of 20±1°C. The semi-static system, renewal at 24 hours for the test. In the preliminary tests the solubility of the test item to dilution water was not reproducible, and the concentration in dispersed suspension containing the test item was not detected. So the test concentration was decided from the concentration which was expected to maintain the concentration in test solution prepared with N,N-Dimethylfomamide (DMF) during exposure and be more than the solubility.   The measured concentrations of the test item in the test solution were 66.5 and 79.9% of the nominal concentration at the preparation and after the renewal, and 23.5 and 40.0% before the renewal and at the end of exposure, respectively. The results were estimated based on the time-weighted mean of the measured concentrations, 0.0495 mg/L, because the measured values of the test item concentration in the test solution were not kept within ±20% of the nominal concentration.   Both the 24-hour and 48-hour EC50 (Median Effective Concentration) of FP-100 were >0.0495 mg/L. No abnormal effects on the test organisms were observed at the exposure concentration.   Consequently, both the highest test concentration which caused 0% immobility and the NOEC (No Observed Effect Concentration) after 48-hours exposure were 0.0495 mg/L. The measured concentration of the test item of the test solution was not kept within ±20% of the nominal concentration during the test. However, it was considered that the concentration of the test item in the test solution was maintained higher than the solubility to the test medium expected from the measured values at 24 hours after preparation of the test solution. Therefore, it is concluded that the test item has no potential for adverse effects on the test organisms near the solubility of the test item to the test medium.

Description of key information

A 48-hour acute Immobilization test of FP-100 with Daphnia magna was conducted.
The results were estimated based on the time-weighted mean of the measured concentrations, 0.0495 mg/L, because the measured values of the test item concentration in the test solution were not kept within ±20% of the nominal concentration.
Both the 24-hour and 48-hour EC50 (Median Effective Concentration) of FP-100 were >0.0495 mg/L.
No abnormal effects on the test organisms were observed at the exposure concentration.
Consequently, both the highest test concentration which caused 0% immobility and the NOEC (No Observed Effect Concentration) after 48-hours exposure were 0.0495 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

0.05 mg/L

Additional information

A 48-hour acute Immobilization test of FP-100 with Daphnia magna was conducted.   Twenty test organisms were used in one exposure level (nominal concentration: 0.100 mg/L), a vehicle control and a control. The water temperature was maintained within the range of 20±1°C. The semi-static system, renewal at 24 hours for the test. In the preliminary tests the solubility of the test item to dilution water was not reproducible, and the concentration in dispersed suspension containing the test item was not detected. So the test concentration was decided from the concentration which was expected to maintain the concentration in test solution prepared with N,N-Dimethylfomamide (DMF) during exposure and be more than the solubility.   The measured concentrations of the test item in the test solution were 66.5 and 79.9% of the nominal concentration at the preparation and after the renewal, and 23.5 and 40.0% before the renewal and at the end of exposure, respectively. The results were estimated based on the time-weighted mean of the measured concentrations, 0.0495 mg/L, because the measured values of the test item concentration in the test solution were not kept within ±20% of the nominal concentration.   Both the 24-hour and 48-hour EC50 (Median Effective Concentration) of FP-100 were >0.0495 mg/L. No abnormal effects on the test organisms were observed at the exposure concentration.   Consequently, both the highest test concentration which caused 0% immobility and the NOEC (No Observed Effect Concentration) after 48-hours exposure were 0.0495 mg/L. The measured concentration of the test item of the test solution was not kept within ±20% of the nominal concentration during the test. However, it was considered that the concentration of the test item in the test solution was maintained higher than the solubility to the test medium expected from the measured values at 24 hours after preparation of the test solution. Therefore, it is concluded that the test item has no potential for adverse effects on the test organisms near the solubility of the test item to the test medium.