Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12-2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
other: TCCA(Toxic Chemical Control Act)- Test Guidelines described in Annex 5 of TCCA- Good Laboratory Practice Standards and Test Guidelines” Notification No. 2012-23, National Institute of Environmental Research, Korea (Aug. 22, 2012)
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
Test substance name: MI RAMER M1086
Appearance : Clear liquid
Manufacturing date: 2012-03-29
Supplier:Miwon Specialty Chemical Co., Ltd.
Receipt day: 2012-05-23
Deli very amount: 399.223 g(Gross)
Cas No.:117646-83-0
Lot No.:120329JY1
Purit y: 95.55 %
Expiration date: 2013-03-28 (Manufactured after 1 year)
Storage condition: Room temperature [(15 ~ 25)°C

Sampling and analysis

Analytical monitoring:
yes

Test organisms

Test organisms (species):
Oryzias latipes
Details on test organisms:
Ricefish (Oryzias latipes) obtained from National Institute of Environmental Research (Environmental Research Complex, Kyungseo-dong, Seo-gu, Incheon, Korea) on May 14, 2007.

Breeding method
Upon receipt, suitable male and female fish, at a ratio of 3 to 2 respectively, without any visible abnormalities and of similar size were selected and placed in a 50 L glass breeding chamber containing maintenance water. Approximately 25 fish (±10%) were bred.
Eggs were harvested from the breeding chambers and placed in hatching chambers. After hatching, the fry were placed in a breeding chamber at 23±2°C and bred. Approximately 30% of the holding water was exchanged once a week. The holding room was artificially illuminated; 16 hours light, 8 hours dark. Fish were fed Brine Shrimp (Ocean Star International, Inc., U.S.A.) in the morning and Top Meal (Jaeilfeed Co., Korea) in the afternoon, approximately 3% of their body weight daily, except from Saturday afternoon to Sunday, when they were fasted.

Acclimation
Prior to initiation, fish without any visible abnormalities and of similar size were acclimated for 10 days in maintenance water.
During the acclimation period, the water temperature and dissolved oxygen concentration in maintenance water were maintained at 22.5–23.2oC and 96.2–99.9% respectively. The room was artificially illuminated with 16 hours of light and 8 hours of dark. The fish were fed Brine Shrimp (Ocean Star International, Inc., U.S.A.) in the morning and Top meal (Jaeilfeed Co., Korea) in the afternoon, at an amount of approximately 3% of their body weight, once daily except from Saturday afternoon to Sunday when they were fasted.
Mortalities were observed from 48 hours after acclimation. During the seven days prior to exposure, the mortality of a batch of fish was less than 5% of the population, thus a batch of fish was used.

Selection
Following acclimation, 5 fish were randomly selected for a range finding study and 10 fish were selected for the definitive study. The fish were blotted dry with a soft tissue and their size was measured with a vernier caliper to confirm a size of 2.0±1.0 cm. As a result, the sizes of the fish were 2.72±0.14 cm and 2.65±0.11 cm (mean±SD) in the range finding study and definitive study, respectively. Weights were recorded (Sartorius, Germany). Consequently, the weights of fish were 0.31±0.01 g and 0.29±0.02 g (mean±SD) in the range finding study and in definitive study, respectively.

Study design

Test type:
semi-static
Water media type:
freshwater

Test conditions

Test temperature:
23±1°C
pH:
6.0–8.5
Dissolved oxygen:
dissolved oxygen level (at least 60% of the air-saturation value 8.53 mg/L (22°C)
Details on test conditions:
Materials and Apparatus
Glass breeding chamber (600W×300D×400H (mm))
Glass test chamber (5 L beaker) (called ‘test chamber’)
Vernier caliper (Digital type, Japan)
Digital thermometer and pH meter (Portable pH meter, Radiometer, France)
Dissolved oxygen meter (Portable DO meter, HACH, U.S.A.)
Electronic balance (BP310S, BP3100S and CP3202S, Sartorius, Germany)

In the range finding study, the mortality of ricefish was 100% at 10 mg/L, while it was 0% at 1 mg/L. Therefore, the highest concentration in the definitive study was determined as 10 mg/L and divided by a geometric ratio of 1.8 to produce additional 4 lower concentration levels including 1, 1.7, 3.1 and 5.6 mg/L. The control group (including the solvent control group) was set.

The volume of test solution was prepared at 3 L in each test chamber. The temperature (23±1°C), dissolved oxygen level (at least 60% of the air-saturation value 8.53 mg/L (22°C)) and pH (6.0–8.5) were checked. Then 10 fish were placed in each chamber for 96 hours of continuous exposure. Biomass to solution and loading ratios did not exceed 1.0 g/L of solution. The temperature of dilution water was maintained at 23±1oC and the room was artificially illuminated with fluorescent lighting; 16 hour light, 8 hour dark cycle. Fish were not fed 24 hours prior to exposure.
One replicate was prepared for the treatment groups and control group (including the solvent control group).
Reference substance (positive control):
yes
Remarks:
A positive control 96 hour study using Oryzias latipes to exposure in Copper (II) Sulfate (Wako Pure Chemical Industries, Ltd.) as a reference material was performed once every six months and an LC50 was determined.

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
> 0.065 mg/L
Nominal / measured:
meas. (geom. mean)
Key result
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 0.065 mg/L
Nominal / measured:
meas. (geom. mean)
Key result
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
0.059 mg/L
Nominal / measured:
meas. (geom. mean)
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.052 mg/L
Nominal / measured:
meas. (geom. mean)

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Based on the results of this study, the LC 50 obtained under the test conditions with the test substance, MIRAMER M1086, was determined to be >0.0651 mg/L (measured concentration) at 24 and 48 hours, 0.059 mg/L (measured concentration, 95% confidence limits: 0.052–0.070 mg/L) at 72 hours and 0.052 mg/L (measured concentration, 95% confidence limits: 0.048–0.057 mg/L) at 96 hours, respectively.
At 96 hours after exposure, the highest concentration causing no mortality and the lowest concentration causing 100% mortality were determined to be 0.0389 and >0.0651 mg/L (measured concentration), respectively.
Executive summary:

Based on the results of this study, the LC 50 obtained under the test conditions with the test substance, MIRAMER M1086, was determined to be >0.0651 mg/L (measured concentration) at 24 and 48 hours, 0.059 mg/L (measured concentration, 95% confidence limits: 0.052–0.070 mg/L) at 72 hours and 0.052 mg/L (measured concentration, 95% confidence limits: 0.048–0.057 mg/L) at 96 hours, respectively.

At 96 hours after exposure, the highest concentration causing no mortality and the lowest concentration causing 100% mortality were determined to be 0.0389 and >0.0651 mg/L (measured concentration), respectively.