[No public or meaningful name is available]

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

December 14 2020 to December 18 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

copper(II) 3,3',3'',3'''-(29H,31H-phthalocyanine-2(or 3),9(or 10),16(or 17),23(or 24)-tetrayltetrasulfonyl)tetrapropane-1-sulfonate and its amides with (2-hydroxypropyl)amine, sodium salts

Molecular formula:

Cannot be adequately assigned

IUPAC Name:

copper(II) 3,3',3'',3'''-(29H,31H-phthalocyanine-2(or 3),9(or 10),16(or 17),23(or 24)-tetrayltetrasulfonyl)tetrapropane-1-sulfonate and its amides with (2-hydroxypropyl)amine, sodium salts

Test material form:

solid: particulate/powder

Details on test material:

Molecular formula: C50H52CuN10Na2O20S8 (main component)
Molecular weight: 1479.02 (main component)
Purity of test item: 93.1%
Impurity
Water: 6.5%
EDTA-4Na: 0.4%
Supplier: FUJIFILM Corporation
Lot number: M-002P
Storage conditions: Store in the dark at room temperature.

Specific details on test material used for the study:

Not specified

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The test solution for analysis was taken out from the middle layer of the test vessel.
Volume of sample: Approximately 10 mL (all test levels)

Test solutions

Vehicle:

yes

Details on test solutions:

The weighed test sample (0.301 g) was mixed with dilution water (2.8 L) in the test vessel, stirred, and dissolved to prepare the test solution of 100 mg/L (upper limit concentration of the test guideline) as a limit test and a control.

Test organisms

Test organisms (species):

Oryzias latipes

Details on test organisms:

Test organism: Medaka ( Oryzias latipes)
Reason for selection of species: Species recommended in the test guideline.
Supplier: CERI Kurume (in-test facility production).
Size: Total length 1-2 cm
Allocation: Random sampling
Hatching date: October 20, 2020 (age at the start of exposure; one-month-old)
Water : Dechlorinated tap water
Acclimation type: Flow-through regime.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

Mortality and performance status were observed at 2, 5 and 24 hours after the start of exposure at 0-1 days after, twice a day ( a.m. and p.m.: 48, 54, 72 and 78 hours after the start of exposure) at 2-3 days after.

Post exposure observation period:

Not specified.

Test conditions

Hardness:

44.1 mg CaCO3/L

Test temperature:

23.9 - 24.1°C

pH:

7.8 - 7.9

Dissolved oxygen:

8.5 - 8.7 mg/L

Nominal and measured concentrations:

The measured concentration of the test item in the test solution at the start of exposure was 102 mg/L.
The measured concentrations of the test item were maintained within the range of 80-120% of the nominal concentration.

Details on test conditions:

Dilution water
Dechlorinated tap water, aerated sufficiently and temperature-controlled, was used. It was confirmed that the concentration of residual chlorine in dilution water was ,:SO. 02 mg/L at the use. The result of chemical characteristics of dilution water met the standard that was provided with the standard operating procedure of this test facility.

Test apparatus and equipment
Test vessel : 3 L glass tank ( diameter: 16 cm, depth: 17 cm)
Cover on test vessel: Transparent plastic lid
Water bath: Plastic tank
Warming/cooling unit RX-401HPN-Z (IW AKI)

Preparation of test solution
The weighed test sample (0.301 g) was mixed with dilution water (2.8 L) in the test vessel, stirred, and dissolved to prepare the test solution of 100 mg/L.

Test condition
Type of test: Static regime (no renewal of test solution)
Exposure duration: 96 hours
Test concentration: 100 mg/L as nominal concentration (limit test as upper limit concentration of the test guideline)
The test concentration was decided from the results of the preliminary study.
The results of the preliminary study are shown in Additional data.
Control: Dilution water without the test item
Replicate: 1 replicate/test level
Number of organism: 7 fish/test level
Volume of test solution: Approximately 2. 8 L/test level
Temperature of test solution: 24±1°C
Aeration: Conducted gently
pH adjustment: No adjustment
Lighting condition: Room light, 16-hour light/8-hour dark (illuminance: 540-1000 lx)
Feeding : No feeding

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The 96-hour LC50 of the test item for Medaka was > 100 mg/L.
Mortality
Number of dead fish in the control at the end of exposure was 0, which met the criterion for the validity of the test (i.e. not exceed one fish).

Observed performance status etc.
No abnormal response was observed in the control.
The following result of observation was based on the comparison with the control organisms. No abnormal response in the exposure level during exposure was observed.

Size of test organism
Total length 1.9±0.1 cm (1.8-2.0 cm)
Body weight 0.063±0.009 g (0.049-0.080 g)

Observation and measurement of test solution
Appearance of test solution
The test solution in the exposure level was blue and clear at the start and end of exposure. The test solution in the control was colourless and clear at the start and end of exposure.

Condition of test solution
Dissolved oxygen concentration, pH and temperature
The measured values of dissolved oxygen concentration, pH and temperature (measured by thermometer of glass stick type) during exposure were 8.5-8.7 mg/L, 7.8-7.9 and 23.9- 24.1 °C, respectively. The measured values of dissolved oxygen concentration met the criterion for the validity of the test ( at least 60% of saturate concentration* at the water temperature).

Total hardness
The measured total hardness in dilution water at the start of exposure was 44.1 mg CaCO3/L.

Illurninance
The illuminance at the top of each test vessel measured at the start of exposure 63 7 and 830 lx, which was within the range specified by the test method (540-1000 lx).

Concentration of test item in test solution
The measured concentration of the test item in the test solution at the start of exposure was 102 mg/L (102% of the nominal concentration), and those at the end of exposure was 106 mg/L (106% of the nominal concentration). The measured concentrations of the test item were maintained within the range of 80-120% of the nominal concentration.

LC50
The 96-hour LC50 of the test item for Medaka was > 100 mg/L.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

This study was conducted as a limit test in order to confirm the effect of the test item on the test organisms at the upper limit concentration of the test guideline.
As a result, no adverse effect on the test organisms was found in the definitive study, and the 96-hour LC50 was > 100 mg/L. The test item concentration in the test solution was maintained within 80-120% of the nominal concentration.
The environmental conditions were within the suitable range. Therefore, it is concluded that this study complied with the applied test guideline.

Executive summary:

The objective of this study is to detemune the 96-hour median lethal concentration (LC50) by conducting an acute toxicity study of the Dye-2019 in Medaka.
Test guideline
OECD Guidelines for Testing of Chemicals, No. 203, June 18, 2019, "Fish, Acute Toxicity Testing"
Test condition: Test organism:Medaka ( Oryzias latipes), Dilution water: Dechlorinated tap water, Test level:  100 mg/L (upper limit concentration of the test guideline) as limit test and a control. The test sample and dilution water were mixed, stirred and dissolved to prepare the test solution. The test concentration was 100 mg/L as nominal concentration (limit test as upper limit concentration of the test guideline).The test concentration was decided from the results of the preliminary study. The test system was a static regime for a 96 hour duration. 7 fish/test level were assessed, the fish were not feed 24 hours prior to the begining of the assessment.

Mortality and performance status were observed at 2, 5 and 24 hours after the start of exposure at 0-1 days after, twice a day ( a.m. and p.m.: 48, 54, 72 and 78 hours after the start of exposure) at 2-3 days after. Since the exposure started in a.m., the observation at 4 days after was only observed at 96 hours after the start of exposure. A fish was considered as dead when the observable motion (motion of mouth and opercula etc.) was not observed and touching of the caudal peduncle with a glass rod caused no reaction. In addition, surviving organisms at the end of exposure were euthanized with anesthesia by using F Al 00 which was an anesthetic for fish (DS Pharma Animal Health).

Analysis of the concentration of the test solution was carried out by HPLC analysis both ath the start and end of the exposure period.

The 96 hour LC50 of the test item was determined to be >100 mg/L, this concentration is based on the nominal concentration.