2-[(1-amino-9,10-dihydro-4-hydroxy-9,10-dioxo-2-anthryl)oxy]ethyl phenyl carbonate

Toxicity to aquatic plants other than algae

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic plants other than algae

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-10-23 to 2014-11-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 221 (Lemna sp. Growth Inhibition Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Duplicate samples from the freshly prepared test media of the only test concentration and the control 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 concentration during the test period, duplicate samples were taken at the end of the test from all test media and the control of the actual test by pouring together the contents of each treatment. Additionally, duplicate samples of the only test concentration at test start and test end were taken and diluted by a factor of 2 with acetonitrile directly after sampling.

Vehicle:

no

Details on test solutions:

Test Concentrations: Filtrate of nominal 100 mg test item/L and a control.
Control: In the control, test medium was used without addition of the test item.
Dosage of Test Item: The test item was not well soluble in test water. In this study, only the inhibitory effects of dissolved test item was tested and thus no concentrations above the solubility limit of the test item in test water were tested. Therefore, a stock suspension of 100 mg test item/L was prepared by suspending 157.1 mg test item in 1571 mL test water for preparing the test concentrations. The stock suspension was stirred for 24 hours in the dark to dissolve as much test item as possible. Then, the undissolved test item was separated by filtration (cellulose nitrate filter). The test medium was prepared just before introduction of the Lemna (= start of the test).
Appearance of the Test Item in Test Medium:
There were no remarkable observations.

Test organisms (species):

Lemna gibba

Details on test organisms:

Species: Lemna gibba G 3
Origin: The fronds introduced in the test were taken from IBACON's in-house laboratory culture.
Breeding Conditions: The plants are cultured in the laboratories of IBACON under standardised conditions according to the test guidelines.
Pre-Culture: For at least 7 days under test conditions with weekly media exchange.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

7 d

Test temperature:

Water Temperature: The temperature was measured daily in a test vessel filled with water and incubated under the same conditions as the test vessels. 22 to 25 °C.

pH:

pH-Values: The pH-value of each treatment was measured at test start and at test end.
7.6 to 8.0 at test start.
8.8 at test end.

Nominal and measured concentrations:

Test Concentrations: Filtrate of nominal 100 mg test item/L and a control.
Control: In the control, test medium was used without addition of the test item.

Details on test conditions:

Test Environment: Controlled environment room
Water Temperature: The temperature was measured daily in a test vessel filled with water and incubated under the same conditions as the test vessels.
22 to 25 °C
pH-Values: The pH-value of each treatment was measured at test start and at test end.
7.6 to 8.0 at test start; 8.8 at test end
Light Regime: Continuous illumination
Light Intensity: 7997 Lux (mean value) with a range of 7630 to 8270 lux
The illumination was achieved by fluorescent tubes, installed above the test vessels. Measurements were performed once during the test at 6 places distributed over the experimental area at the surface of the test media. Differences in light intensity over the test area did not exceed ±15 %.

Reference substance (positive control):

yes

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Key result

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

7 d

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Duration:

7 d

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

7 d

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

7 d

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

7 d

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Duration:

7 d

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

7 d

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

7 d

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Details on results:

Biological Results
Growth Inhibition: The 7-day EyC50 was calculated to be > 100 mg test item/L for frond number and dry weight.
The 7-day ErC50 was calculated to be >100 mg test item/L for frond number and dry weight.
The 7-day EyC10 was calculated to be >100 mg test item/L for frond number and dry weight.
The 7-day ErC10 was calculated to be >100 mg test item/L for frond number and dry weight.
The 7-day NOEyC and the LOEyC were determined to be 100 and >100 mg test item/L for frond number and dry weight, respectively.
The 7-day NOErC and the LOErC were determined to be 100 and >100 mg test item/L for frond number and dry weight, respectively.
Shape of Fronds: The shape of fronds and colonies after the test period of 7 days in the nominal test concentration of 100 mg test item/L was not different to those in the control.

Analytical Results:
At the start of the test and after 7 days test duration all values were below the Limit of Detection.
Therefore, all reported results refer to nominal concentrations.

Results with reference substance (positive control):

Results of the most recent Test with the Reference Item 3,5 Dichlorophenol performed in August to September 2014 (IBACON Project No.: 88302240)
Parameter Yield
(frond number) Growth rate
(frond number) Yield
(dry weight) Growth rate
(dry weight)
[mg test item/L] [mg test item/L] [mg test item/L] [mg test item/L]
EC50 (7-day) 6.24 10.0 6.67 9.37
95 % conf. limits 4.87 - 7.99 7.82 - 12.8 5.55 - 8.06 8.43 - 10.4

EC20 (7-day) 3.43 6.68 4.48 6.46
95 % conf. limits 1.96 - 4.47 3.21 - 8.35 3.02 - 5.41 5.05 - 7.38

EC10 (7-day) 2.50 5.41 3.64 5.32
95 % conf. limits 1.15 - 3.51 1.88 - 7.18 2.10 - 4.60 3.77 - 6.34

7-day NOEC 0.625 1.25 2.5 2.5
7-day LOEC 1.25 2.5 5.0 5.0
Values refer to nominal test concentrations

Reported statistics and error estimates:

The ErC50 and the EyC50 (see Definitions), the corresponding EC20 and EC10 values could not be calculated due to absence of toxicity of the test item.
For the determination of the 7-day LOEyC and NOEyC values significant differences at the test concentrations compared to the control values were tested by the Student-t test (frond number and dry weight).
For the determination of the 7-day LOErC and NOErC values significant differences at the test concentrations compared to the control values were tested by the Student-t test (frond number and dry weight).
The software used to perform the statistical analysis was ToxRat Professional, Version 2.10.05, ToxRat Solutions GmbH.

Influence of FAT 93504/B TE on the Growth of Lemna gibba

Parameter	Yield (frond number) [mg test item/L]	Growth rate (frond number) [mg test item/L]	Yield (dry weight) [mg test item/L]	Growth rate (dry weight) [mg test item/L]
EC50 (7-day)	> 100	> 100	> 100	> 100
95 % conf. limits	n.d.	n.d.	n.d.	n.d.
EC10(7-day)	> 100	> 100	> 100	> 100
95 % conf. limits	n.d.	n.d.	n.d.	n.d.
7-day NOEC	100	100	100	100
7-day LOEC	> 100	> 100	> 100	> 100

n.d.: not determinable
Values refer to nominal test concentrations

Summary of Analytical Results

sample description	% of
[mg/L]	nominal^1	n
control	n.a.	4
Filtrate of 100 mg/L	< LOD	4

¹mean value of all measured samples per treatment group
RSD: relative standard deviation per treatment group
n: number of analysed samples
n.a.: not applicable

Validity criteria fulfilled:

yes

Conclusions:

The influence of FAT 93504/B TE on the growth of the freshwater plant Lemna gibba was assessed in a static dose-response test.
The 7-day EyC50 was calculated to be >100 mg test item/L for frond number and dry weight.
The 7-day ErC50 was calculated to be >100 mg test item/L for frond number and dry weight.
The 7-day NOEyC and the LOEyC were determined to be 100 and >100 mg test item/L for frond number and dry weight, respectively.
The 7-day NOErC and the LOErC were determined to be 100 and >100 mg test item/L for frond number and dry weight, respectively.

Executive summary:

The toxicity of the test item to the aquatic plant Lemna gibba in a growth inhibition test was determined based on the procedures indicated by the following internationally accepted methods:

- OECD 221 equivalent to EU C. 26

Biological Results:

Growth Inhibition:

The 7-day E_yC₅₀was calculated to be >100mg test item/Lfor frond number and dry weight.

The 7-day E_rC₅₀was calculated to be >100 mg test item/Lfor frond number and dry weight.

The 7-day E_yC₁₀was calculated to be >100 mg test item/Lfor frond number and dry weight.

The 7-day E_rC₁₀was calculated to be >100 mg test item/Lfor frond number and dry weight.

The 7-day NOE_yC and the LOE_yC were determined to be100 and >100 mg test item/Lfor frond number and dry weight, respectively.

The 7-day NOE_rC and the LOE_rC were determined to be100 and >100 mg test item/Lfor frond number and dry weight, respectively.

Shape of Fronds:

The shape of fronds and colonies after the test period of 7 days in the nominal test concentration of 100mg test item/Lwas not different to those in the control.

Analytical Results: At the start of the test and after 7 days test duration all values were below the Limit of Detection. Therefore, all reported results refer tonominal concentrations.

Conclusion:

The influence of FAT 93504/B TE on the growth of the freshwater plantLemnagibbawas assessed in a static dose-response test.

The 7-day E_yC₅₀was calculated to be >100mg test item/Lfor frond number and dry weight.

The 7-day E_rC₅₀was calculated to be> 100 mg test item/Lfor frond number and dry weight.

The 7-day NOE_yC and the LOE_yC were determined to be100 and >100 mg test item/Lfor frond number and dry weight, respectively.

The 7-day NOE_rC and the LOE_rC were determined to be100 and >100 mg test item/Lfor frond number and dry weight, respectively.

This study is classified acceptable and satisfies the guideline requirements for aquatic plant studies.

Description of key information

The influence of FAT 93504/B on the growth of the freshwater plant Lemna gibba was assessed in a static dose-response test, and the results are as follows:    

The 7-day EyC50 was calculated to be >100 mg test item/L for frond number and dry weight.

The 7-day ErC50 was calculated to be >100 mg test item/L for frond number and dry weight.

The 7-day NOEyC and the LOEyC were determined to be 100 and >100 mg test item/L for frond number and dry weight, respectively.

The 7-day NOErC and the LOErC were determined to be 100 and >100 mg test item/L for frond number and dry weight, respectively.

Key value for chemical safety assessment

EC50 for freshwater plants:

100 mg/L

EC10 or NOEC for freshwater plants:

100 mg/L

Additional information

The toxicity of the test item to the aquatic plant Lemna gibba in a growth inhibition test was determined based on the procedures indicated by the following - OECD 221 equivalent to EU C. 26

The influence of FAT 93504/B TE on the growth of the freshwater plant Lemna gibba was assessed in a static dose-response test.

The 7-day EyC50 was calculated to be >100 mg test item/L for frond number and dry weight.

The 7-day ErC50 was calculated to be >100 mg test item/L for frond number and dry weight.

The 7-day NOEyC and the LOEyC were determined to be 100 and >100 mg test item/L for frond number and dry weight, respectively.

The 7-day NOErC and the LOErC were determined to be 100 and >100 mg test item/L for frond number and dry weight, respectively.