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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
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)
Version / remarks:
OECD Guidelines for Testing of Chemicals, No.202, April 13, 2004, "Daphnia sp., Acute immobilisation Test"
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Acute Immobilization Test stipulated in the "Testing Methods for New Chemical Substances" (March 31, 2011, No. 0331-7, Pharmaceutical and Food Safety Bureau
Version / remarks:
Acute Immobilization Test stipulated in the "Testing Methods for New Chemical Substances" (March 31, 2011, No. 0331-7, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare; March 29, 2011, No. 5, Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry; No. 110331009, Environmental Policy Bureau, Ministry of the Environment, Japan; April 2, 2012 partial revision, No. 0402-1, Pharmaceutical and Food Safety
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: OECD Guidance Document, No.23, September 2000, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures"
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Chemical name: (Z)-1-chloro-2,3,3,3-tetrafluoropropene
Other name: HCF0-1224yd(Z)
Molecular formula: C3HClF4
Molecular weight: 148.49
Purity of test item: >99% (GC area%)
Supplier: ASAHI GLASS CO., LTD.
Lot number: 151007
The test item was treated as 100% in purity.
Analytical monitoring:
yes
Details on sampling:
Frequency of measurement: At the start of exposure, before and after the renewal, and at the end of exposure.
Sample for measurement: Another solution sampled separately from the preparation container. Then one test vessel in each test level (before the renewal and at the end of exposure)
Volume of sample: Approximately 50 mL (all test levels)
Vehicle:
no
Details on test solutions:
Dilution water
Dechlorinated tap water, aerated sufficiently and temperature-controlled, was used. Some chemical characteristics of the dilution water measured regularly are listed in Appendix 1. The result of chemical characteristics of the dilution water met the standards that were provided with the standard operation procedure of this laboratory.

Test apparatus and equipment
Test vessel: 50 mL stoppered Erlenmeyer flask (closed vessel)
Water bath: Plastic tank. Warming/cooling unit; Type HCA 250 (Sato craft)

Preparation of test solution
After dilution water was added in sampling bag for gas analysis, gaseous test sample was enclosed in the bag. The bag was shaken about 2 hours. After that, the solution was settled more than 12 hours about 22°C to prepare the stock solution. The day that the test solution was prepared, test item concentration of stock solution was measured (Analytical method was shown in Appendix 2.). Based on the result of the measured concentration, required volumes of the stock solution and dilution water
were mixed and stirred in a preparation container to prepare the test solution which was divided into each test vessel.


Test organisms (species):
Daphnia magna
Details on test organisms:
Species: Daphnia magna Clone A
Reason for selection of species: Species recommended in the test guidelines
Source of supply: The University of Sheffield, UK
Date of supply: July 9, 1990
Acclimation: Young daphnids produced by parents that were cultured in this laboratory were used. The parents to obtain young daphnids were bred in the same quality of water ( dechlorinated tap water), water temperature (20± 1 °C) and photoperiod (16-hour light/8-hour dark) as used in the test. The parent animals used for the test were same lot and their age and survival rate were 14-day old and 100%, respectively. Chlorella vulgaris of 0.1-0.2 mgC (Organic carbon content)/day per Daphnia was fed to the parents once a day.
Selection of young daphnids: Less than 24-hour-dd daphnids.
Confirmation of reproducibility of test system: A 48-hour acute immobilization test of a reference substance with the test organisms was periodically conducted. The latest data is shown below.
Allocation to the test groups: Random sampling

Number of organism: 20 daphnids/test level (5 daphnids/test vessel)


Test type:
semi-static
Water media type:
freshwater
Total exposure duration:
48 h
Remarks on exposure duration:
Semi-static regime (renewal at 24 hours after), closed system without headspace
Post exposure observation period:
None
Hardness:
28 mg/l
Test temperature:
20 °C ± 1 °C
pH:
7.7
Dissolved oxygen:
>3 mg/l
Conductivity:
11
Nominal and measured concentrations:
Nominal concentrations :0, 4.27, 9.39, 20.7, 45.5, 100 mg/l
Measured concentrations: Geometric mean - 0, 3.97, 9.35, 20.1, 41.0, 96.9 mg/l
Details on test conditions:
Test conditions

Type of test: Semi-static regime (renewal at 24 hours after), closed system without headspace
Exposure duration: 48 hours
Test concentration: 100, 45.5, 20.7, 9.39, 4.27 mg/L (a geometric series with a factor of 2.2). The test concentrations and the geometric factor were decided from the results of preliminary study.
Control: Dilution water without the test item
Replicate: 4 replicates/test level
Volume of test solution: Approximately 200 mL/test level ( approximately 50 mL/test vessel)
Temperature of test solution: 20±1 °C
Dissolved oxygen concentration: More than 3 mg/L without aeration
pH adjustment: No adjustment
Lighting condition: Room light, 16-hour light/8-hour dark
Feeding: No feeding

Observation of test organisms

Observation about immobility and symptoms of test organism was conducted at 24 and 48 hours after exposure. Daphnids were considered immobile if they were not able to swim within 15 seconds after gentle agitation of the test vessel.

Appearance of test solution

Observation at the start of exposure and before the renewal (at 24 hours after)

Condition of test solutions

Item of measurement: Dissolved oxygen concentration, pH and temperature
Frequency of measurement: At the start of exposure, before and after the renewal, and at the end of exposure
Sample for measurement: Another solution sampled separately from the container for preparation ( at the preparation). One test vessel in each test level (before the renewal and at the end of exposure)
Dissolved oxygen meter: YSI MODEL 58 (YSI Nanotech Japan)
pH meter: HM-21P (DKK-TOA)
Thermometer: Glass stick type
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (JlS special grade, Wako Pure Chemical Industries, Ltd. Lot No. RCK7838)
Duration:
48 h
Dose descriptor:
EC100
Remarks:
immobilisation
Effect conc.:
96.7 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Remarks:
immobilisation
Effect conc.:
3.97 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
The minimum concentration causing 100% immobility at 48 hours was 96.9 mg/L. Maximum concentration causing no immobility at 48 hours was 3.97 mg/L. Immobility in the control during exposure was 0%, which meets the criterion for the validity of the test (i.e. not more than 10%).

The minimum concentration causing 100% immobility at 48 hours was 96.9 mg/L. Maximum concentration causing no immobility at 48 hours was 3.97 mg/L. Immobility in the control during exposure was 0%, which meets the criterion for the validity of the test (i.e. not more than 10% ).

Observation and measurement of test solution

a) Appearance of test solution
The test solutions were colourless and clear at the start and before the renewal.

b) Condition of test solution
The measured values of dissolved oxygen concentration, pH and temperature of the test solutions during exposure ranged from 8.6 to 8.9 mg/L, from 7.8 to 8.0 and from 19.7 to 20.0°C, respectively. The measured values of dissolved oxygen concentration met the criterion for the study validity (more than 3 mg/L at the end of exposure).

c) Concentration of test item in test solution
The measured concentrations of the test item in the test solutions at the start of exposure and after the renewal ranged from 3. 77 to 108 mg/L, and those before the renewal and at the end of exposure ranged from 3.80 to 99 .6 mg/L which were 78.9-103% of the concentrations at the preparation.

The 24-hour and 48-hour EC50s of the test item for Daphnia magna were 69.5 mg/L (95% confidence limits: 56.9-83.7 mg/L) and 18.9 mg/L (95% confidence limits: 14.9-24.0 mg/L), respectively.

This study was conducted in order to estimate the EC50 of the test item on the test organisms below the solubility of the test item in dilution water. As a result, 48-hour ECso was 18.9 mg/L (95% confidence limits: 14.9-24.0 mg/L). Since the test item has volatility, closed vessel was used in this study. In addition, the definitive study was selected semi-static regime renewal at 24 hours after to maintain the test item concentration during exposure. As the results, the test item concentrations in the test solutions were mostly
maintained. The environmental conditions were within the suitable range; therefore, it is concluded that this study complied with the applied test guidelines.
Results with reference substance (positive control):
Reference substance: Potassium dichromate (JlS special grade, Wako Pure Chemical Industries, Ltd. Lot No. RCK7838)
Period of study: October 14- October 16, 2015
48-hour EC50: 0.18mg/L
This value was within the stipulated range (mean± 2S.D.) [mean± S.D.: 0.23±0.06 mg/L (n=94)] to background data in this laboratory.

 Measured Concentration mg/l a

Immobility (%)

24 hours 

48 hours

Replicate

Test Level

Replicate

Test Level

Control

A

0

0

0

0

B

0

0

C

0

0

D

0

0

3.97

A

0

0

0

0

B

0

0

C

0

0

D

0

0

9.35

A

0

0

20

 20

B

0

20

C

0

20

D

0

20

20.1         

A

0

0

40

45

B

0

60

C

0

20

D

0

60

41.0

A

0

5

80

90

B

20

100

C

0

100

D

0

80

96.9

80

85

100

100

B

60

100

C

100 

100

D

100

100

a = Geometric mean of the measured concentrations ( also expressed as measured concentration in the following tables and figure)

Observed Abnormal Response

 Measured Concentration (mg/l)

Observed abnormal response (number of affected test organism)

24 hours

48 hours

Control

None

None

3.97

None

None

9.35

None

Immobilisation (4)

Lethargic (1)

Reduced activity (3)

20.1

None

Immobilisation (9) 

Reduced activity (2)

41.0

Immobilisation (1) 

Reduced activity (2)

Immobilisation (18)

Lethargic (8)

96.9

Immobilisation (17)

Reduced activity (3)

Immobilisation (20)

Lethargic (17)

EC50 to Daphnia magna

Exposure duration

EC50 (mg/L)

95% confidence limits (mg/L) (Slope of the concentration-response curve)

Statistical procedure used for determination of EC50. 

24-hour

69.5

59.9-83.7

Probit analysis

48-hour

18.9

14.9-24.0

Probit analysis

 

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of HCFO-1224yd(Z) to daphnia was determined in a 48-hour test according to OECD guideline 202. The EC50 (immobilisation) was determined to be 18.9 mg/l (Geometric mean measured concentration). There were no deviations from the guideline and the validation criteria were all met.
Executive summary:

The acute toxicity of HCFO-1224yd(Z) to daphnia was determined in a 48-hour test according to OECD guideline 202. The EC50 (immobilisation) was determined to be 18.9 mg/l (Geometric mean measured concentration). There were no deviations from the guideline and the validation criteria were all met.

Description of key information

The acute toxicity of HCFO-1224yd(Z) to daphnia was determined in a 48-hour test according to OECD guideline 202. The EC50 (immobilisation) was determined to be 18.9 mg/l (Geometric mean measured concentration). There were no deviations from the guideline and the validation criteria were all met.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
18.9 mg/L

Additional information