Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Ecotoxicological information

Toxicity to aquatic algae and cyanobacteria

Currently viewing:

Administrative data

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April 06, 2018 to April 25, 2018
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
GLP compliance:
yes
Remarks:
The range-finding test was not performed in compliance with the GLP-Regulations and is excluded from the Statement of Compliance.

Test material

Constituent 1
Chemical structure
Reference substance name:
reaction mass of (1R,3R)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylic acid and (1S,3S)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylic acid
EC Number:
614-283-9
Cas Number:
68127-59-3
Molecular formula:
C9H10ClF3O2
IUPAC Name:
reaction mass of (1R,3R)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylic acid and (1S,3S)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylic acid
Test material form:
solid

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: The concentration of the test item in the treatement samples were determined as followed:
Quantification:Samples were quantified by measuring the peak area with reference to the calibration curve. The latter was obtained by correlation of peak area of the standard solutions to their corresponding concentration. The correlation was performed using a linear regression function given by equation (1):
y = a*x + b (1)
where
y: peak area
x: concentration of analyte
a: slope
b: y-axis intercept
Calculated Concentration: The concentration of the analyte in the treatment samples and in the control sample was calculated by equation (2):
c = x * d (2)
where
c: concentration of analyte in original sample
x: concentration of analyte in the analysed sample (calculated in equation (1))
d: dilution factor

- Sampling method: The samples were taken from the biological phase of the study. Duplicate samples from the freshly prepared test media (containing algae) of all test concentrations and from the controls were taken at the start of the test. For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the test period, duplicate samples from the test media of all test concentrations and the controls (containing algae) were taken at the end of the test (after the 96 hours test period) by pouring together the contents of the test beakers of each treatment. The samples were diluted by a factor of two with acetonitrile. One additional sample of the control and of the dilution solvent was taken at test start and test end without any sample treatment. The concentrations of the test item were analysed in one of the duplicate test media samples from all test concentrations and the control from both sampling times (0 and 96 hours).

- Sample storage conditions before analysis: All samples were stored in a freezer (≤ - 20 °C), protected from light until analysis was performed. Afterwards the samples were again stored deep frozen (≤ -20 °C) and will be kept stored up to the date of the final report.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 24 hours before test start a suspension of the highest nominal test concentration of 300 mg/L was prepared by suspending 199.5 mg test item in 665 mL. The suspension was adjusted to a pH of 8.1 using 1 M NaOH and was stirred for 24 hours in the dark to dissolve as much test item as possible. Then, non-dissolved fractions of the test item were separated from the test medium by membrane filtration (0.45 µm cellulose acetate filter). Adequate volumes of this filtrate were mixed into test water to prepare the test media for the desired 1:3.2, 1:10, 1:32, 1:100 dilutions.
The test media were prepared just before introduction of the algae (= start of the test).
- Controls: In the control, test water was used without addition of the test item.

Test organisms

Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Common name: Raphidocelis subcapitata (KORSHIKOV)
- Strain: Strain No. 61.81 SAG, formely know as Selenastrum capricornutum and Pseudokirchneriella subcapitata (KORSHIKOV)
- Source: The algae were supplied by the "Sammlung von Algenkulturen, Albrecht-von-Haller-Institut für Pflanzenwissenschaften, Universität Göttingen", 37073 Göttingen, Germany.
- Method of cultivation: The algae were cultivated in the laboratories of ibacon under standardised conditions of the test guidelines. For the evaluation of the quality of the algae and the experimental conditions the reference item potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions. Last test was perfomed in January 2018.

ACCLIMATION
- Acclimation period: Cells to start the test were taken from an exponentially growing pre-culture, which was set up 3 days prior to the test start under the same conditions as in the test.
- Culturing media and conditions:
Water Temperature: The temperature was measured daily in an Erlenmeyer flask filled with water and incubated under the same conditions as the test flasks. Water temperature was: 23.0 to 23.8 °C
pH-Values: The pH was measured in all test item concentrations and the control at the start and the end of the test. pH was: 8.0 to 8.2 at test start and 8.0 to 10.2 at test end
Light Regime: Continuous illumination
Light Intensity: The light intensity was measured once during the test at 6 positions distributed over the experimental area at the surface of the test media.
Mean light intensity: 7730 lux (range: 6870 to 8620 lux)
Recording: Test conditions were recorded with suitable instruments and documented in the raw data.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
0.24 mmol/L (= 24 mg/L) as CaCO3
Test temperature:
23.0 to 23.8°C
pH:
8.0 to 8.2 at test start and 8.0 to 10.2 at test end
Conductivity:
<5 µScm-1
Nominal and measured concentrations:
nominal concentration: 300 mg test item/L and dilutions of this filtrate of 1:3.2, 1:10, 1:32, 1:100 equivalent to the following mean measured concentrations: 189, 52.0, 17.7, 5.40 and 1.65 mg test item/L.
Details on test conditions:
TEST SYSTEM
- Test vessel: Erlenmeyer flasks of 50 mL volume
- Type: closed; the flasks were covered with glass disehs and incubated in a water batch
- Material: Erlenmeyer flasks of 50 mL volume with 50 mL of test medium
- Initial cells density: 5000 algal cells per mL test medium
- Control end cells density: 288.285*[10000/mL] algal cells per mL test medium after 96 hours
- No. of vessels per concentration (replicates): three replicates per test concentration
- No. of vessels per control (replicates): six replicates in the control

GROWTH MEDIUM
- Standard medium used: yes (reconstituted water, OECD medium)

TEST MEDIUM / WATER PARAMETERS
Macro-nutrients: NaHCO3 50.0 mg/L
CaCl2 x 2 H2O 18.0 mg/L
NH4Cl 15.0 mg/L
MgSO4 x 7 H2O 15.0 mg/L
MgCl2 x 6 H2O 12.0 mg/L
KH2PO4 1.6 mg/L
Trace Elements: Na2EDTA x 2 H2O 100.0 µg/L FeCl3 x 6 H2O 64.0 µg/L
MnCl2 x 4 H2O 415.0 µg/L
H3BO3 185.0 µg/L
Na2MoO4 x 2 H2O 7.0 µg/L
ZnCl2 3.0 µg/L
CoCl2 x 6 H2O 1.5 µg/L
CuCl2 x 2 H2O 0.01 µg/L

- Alkalinity: pH 8.1 ± 0.1
- Ca/mg ratio: 0.24 mmol/L (=24 mg/L) as CaCO3

OTHER TEST CONDITIONS
- Adjustment of pH: yes with 1M NaOH to pH 8.1 ± 0.1
- Light regime:Continuous illumiation
- Light intensity and quality: The light intensity was measured once during the test at 6 positions distributed over the experimental area at the surface of the test media.Mean light intensity: 7730 lux (range: 6870 to 8620 lux)

EFFECT PARAMETERS MEASURED
- Determination of cell concentrations: The cell density on each observation time was determined by spectrophotometric measurement.

TEST CONCENTRATIONS
- Range finding study: For the determination of the test concentrations a range-finding test was performed. The range-finding test was not performed in compliance with the GLP-Regulations and is excluded from the Statement of Compliance. The concentrations tested were filtrates of 300 mg test item/L and dilutions of 1:10, 1:100 and 1:1000.
- Test concentrations: The concentrations tested for the definitive test were a filtrate of 300 mg test item/L and dilutions of this filtrate of 1:3.2, 1:10, 1:32, 1:100 equivalent to the following mean measured concentrations: 189, 52.0, 17.7, 5.40 and 1.65 mg test item/L. Additionally a control was tested.
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
61.9 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
155 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EC20
Effect conc.:
28.8 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EC20
Effect conc.:
83 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
19.3 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
59.8 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
1.65 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
5.4 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
5.4 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
17.7 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Key result
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
72.9 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Key result
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
> 189 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Key result
Duration:
96 h
Dose descriptor:
EC20
Effect conc.:
40.8 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Key result
Duration:
96 h
Dose descriptor:
EC20
Effect conc.:
108 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Key result
Duration:
96 h
Dose descriptor:
EC10
Effect conc.:
30.2 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Key result
Duration:
96 h
Dose descriptor:
EC10
Effect conc.:
80.1 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
1.65 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
5.4 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Key result
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
5.4 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Key result
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
17.7 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Details on results:
- Exponential growth in the control: yes
The microscopic examination of the shape of the algal cells after 96 hours of test duration did not show any difference between the algae that had been growing at a nominal test concentration of 189 mg test item/L and the algal cells in the control. Thus, the shape of the algal cells was not obviously affected up to this test concentration, the highest concentration tested. Regarding the appearance of the test item in the test media, there were no remarkable observations at each concentration tested.
Reported statistics and error estimates:
Based on the calculated cell densities, the 72 and 96-hour ErC50, EbC50 and EyC50, the corresponding EC20 and EC10 values and where possible their 95 %-confidence limits were calculated by Probit analysis.
For the determination of the 72 and 96-hour LOEC and the 72 and 96-hour NOEC, the calculated growth rates, biomass and yields at each test concentration were tested for significant differences compared to the control values by the Williams t-test (growth rate and yield and area under the growth curve).
The software used to perform the statistical analysis was ToxRat Professional, Version 3.2.1, ToxRat Solutions GmbH.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The influence of the test item on the growth ofthe freshwater green algae Raphidocelis subcapitatawas assessed in a static concentration-response test.The 96-hour ErC50 value was calculated to be >189 mg test item/L, the 96-hour EbC50was calculated to be 72.9 mg test item/L and the 96-hour EyC50 was calculated to be 83.8 mg test item/L. The 96-hour NOErC was determined to be 5.40 and the associated 96-hour LOErC was 17.7 mg/L. The 96-hour NOEbC was determined to be 1.65 and the associated 96-hour LOEbC was 5.40 mg/L. The 96-hour NOEyC was determined to be 5.40 ml/L and the associated 96-hour LOEyC was 17.7 mg/L.
Executive summary:

The purpose of this test was to determine the inhibitory effect of the test item on the growth of the freshwater green algae Raphidocelis subcapitata. Exponentially growing cultures of this unicellular green algal species were exposed toa filtrate of 300 mg test item/L and dilutions of this filtrate of 1:3.2, 1:10, 1:32 and 1:100 equivalent to the following mean measured concentrations: 189, 52.0, 17.7, 5.40 and 1.65 mg test item/L, alongside a control under defined conditions. The inhibition of growth in relation to control cultures was determined over a test period of 96 hours, and thus over several algal generations.

The results are presented in the table here below:

 

72 hour

96 hours

Parameter

Biomass

Growth rate

Yield

Biomass

Growth rate

Yield

EC50[mg/L]

61.9

155

62.5

72.9

> 189

83.8

95 % CI

57.9 - 66.4

150 -161

57.5 -68.2

68.5 - 77.9

n.d.

78.3 - 90.8

 

 

 

 

 

 

 

EC20[mg/L]

28.8

83.0

29.0

40.8

108

56.0

95 % CI

25.8 - 31.6

77.2 -88.3

25.3 - 32.5

37.4 - 44.0

102 - 114

52.5 - 59.4

 

 

 

 

 

 

 

EC10[mg/L]

19.3

59.8

19.4

30.2

80.1

45.3

95 % CI

16.6 - 21.8

54.1 - 65.1

16.1 - 22.5

26.7 - 33.3

73.1 - 86.5

41.8 - 48.5

 

 

 

 

 

 

 

NOEC [mg/L]

1.65

5.40

5.40

1.65

5.40

5.40

LOEC[mg/L]

5.40

17.7

17.7

5.40

17.7

17.7

n.d. = not determinable
Values refer to mean measured test concentrations

The concentrations of the test item were determined in one of the duplicate test media samples from all test concentrations (filtrate of test item stock solution and its dilutions) and in one of the duplicate control samples from all sampling times. The mean measured values (rounded to three significant digits) are

189 mg test item/L in the filtrate,

52.0 mg test item/L in the 1:3.2 dilution of filtrate,

17.7 mg test item/L in the 1:10 dilution of filtrate,

5.40 mg test item/L in the 1:32 dilution of filtrate and

1.65 mg test item/L in the 1:100 dilution of filtrate.

The influence of the test item on the growth ofthe freshwater green algae Raphidocelis subcapitata was assessed in a static concentration-response test.The 96-hour ErC50 value was calculated to be >189 mg test item/L, the 96-hour EbC50 was calculated to be 72.9 mg test item/L and the 96-hour EyC50 was calculated to be 83.8 mg test item/L. The 96-hour NOErC was determined to be 5.40 and the associated 96-hour LOErC was 17.7 mg/L. The 96-hour NOEbC was determined to be 1.65 and the associated 96-hour LOEbC was 5.40 mg/L. The 96-hour NOEyC was determined to be 5.40 ml/L and the associated 96-hour LOEyC was 17.7 mg/L.