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Diss Factsheets

Ecotoxicological information

Toxicity to aquatic plants other than algae

Administrative data

Endpoint:
toxicity to aquatic plants other than algae
Remarks:
Toxicity to the Aquatic Plant Lemna gibba
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-04-10 to 2017-05-08
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
OECD, EU guideline, GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 221 (Lemna sp. Growth Inhibition Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Commission Regulation (EC) No 761/2009, C.26.
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", December 15, 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

1
Chemical structure
Reference substance name:
disodium 2,4-dihydroxy-5-[(2-hydroxy-3-nitro-5-sulphonato-phenyl)azo]benzoate, reaction products with chromium triacetate
Molecular formula:
not applicable
IUPAC Name:
disodium 2,4-dihydroxy-5-[(2-hydroxy-3-nitro-5-sulphonato-phenyl)azo]benzoate, reaction products with chromium triacetate

Sampling and analysis

Analytical monitoring:
no
Remarks:
No analytical dose verification was performed since the test substance (type UVCB) cannot be analysed by standard methods in such low test concentrations used in this study.

Test solutions

Vehicle:
no
Details on test solutions:
Test Concentrations: 100, 32, 10, 3.2 and 1.0 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 concentrations of 100, 32, 10, 3.2 and 1.0 mg test item/L were prepared separately by dissolving 94.3, 31.5, 9.8, 3.12 and 2.01 mg test item into 943, 984.4, 980, 975 and 2010 mL test water by intense stirring for approximately 24 hours and ultrasonic treatment for 10 minutes.
The test media were prepared just before introduction of the Lemna (= start of the test).

Test organisms

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.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
7 d

Test conditions

Test temperature:
22.2 to 24.9 °C
pH:
7.4 to 7.6 at test start;
8.4 to 8.5 at test end
Nominal and measured concentrations:
nominal: 100, 32, 10, 3.2 and 1.0 mg test item/L and a control.
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.
22.2 to 24.9 °C
pH-Values: The pH-value of each treatment was measured at test start and at test end.
7.4 to 7.6 at test start;
8.4 to 8.5 at test end
Light Regime: Continuous illumination
Light Intensity: 8050 Lux (mean value) with a range of 7130 to 8670 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
Remarks:
For the evaluation of the quality of the Lemna gibba and the experimental conditions the substance 3,5-dichlorophenol is tested at least twice a year to demonstrate satisfactory test conditions.

Results and discussion

Effect concentrationsopen allclose all
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
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:
EC20
Effect conc.:
7.59 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
frond number
Remarks on result:
other: 95 % CL
Duration:
7 d
Dose descriptor:
EC20
Effect conc.:
12.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: 95 % CL
Duration:
7 d
Dose descriptor:
EC20
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: 95 % CL
Duration:
7 d
Dose descriptor:
EC20
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.:
1.13 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
frond number
Remarks on result:
other: 95 % CL
Duration:
7 d
Dose descriptor:
EC10
Effect conc.:
11.56 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: 95 % CL
Duration:
7 d
Dose descriptor:
EC10
Effect conc.:
11.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: 95 % CL
Duration:
7 d
Dose descriptor:
EC10
Effect conc.:
30.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: 95 % CL
Duration:
7 d
Dose descriptor:
NOEC
Effect conc.:
3.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
frond number
Duration:
7 d
Dose descriptor:
NOEC
Effect conc.:
3.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
7 d
Dose descriptor:
NOEC
Effect conc.:
3.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
7 d
Dose descriptor:
NOEC
Effect conc.:
3.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
7 d
Dose descriptor:
LOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
frond number
Duration:
7 d
Dose descriptor:
LOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
7 d
Dose descriptor:
LOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
7 d
Dose descriptor:
LOEC
Effect conc.:
10 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, respectively.
The 7-day ErC50 was calculated to be > 100 mg test item/L for frond number and dry weight, respectively.
The 7-day EyC10 was calculated to be 1.13 and 1.56 mg test item/L for frond number and dry weight, respectively.
The 7-day ErC10 was calculated to be 11.7 and 30.6 mg test item/L for frond number and dry weight, respectively.
The 7-day NOEyC and the LOEyC were determined to be 3.2 and 10 mg test item/L for frond number and dry weight, respectively.
The 7-day NOErC and the LOErC were determined to be 3.2 and 10 mg test item/L for frond number and dry weight, respectively.
Results with reference substance (positive control):
Results of the most recent Test with the Reference Item 3,5 Dichlorophenol performed in January 2017
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) 4.63 6.24 4.77 6.00
95 % conf. limits 4.30 - 4.98 6.07 - 6.42 4.47 - 5.08 5.77 - 6.36

EC20 (7-day) 3.11 4.79 3.41 4.92
95 % conf. limits 2.68 - 3.45 4.64 - 4.93 2.19 - 3.76 4.79 - 5.02

EC10 (7-day) 2.53 4.17 2.87 4.43
95 % conf. limits 2.06 - 2.89 4.00 - 4.32 2.28 - 3.27 4.22 - 4.57

7-day NOEC 2.5 2.5 2.5 2.5
7-day LOEC 5 5 5 5
Values refer to nominal test concentrations

Reported statistics and error estimates:
The ErC50 and the EyC50 (see Definitions), the corresponding EC20 and EC10 values and where possible their 95 %-confidence limits were calculated by Probit analysis.
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 Williams t-test.
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 Williams t-test.
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
Remarks:
Doubling Time of Frond Number in Control: 2.1 days, validity criterion was met.
Conclusions:
The influence of the test item on the growth of the freshwater plant Lemna gibba was assessed in a static concentration-response test.
The 7-day EyC50 was calculated to be > 100 mg test item/L for frond number and dry weight, respectively.
The 7-day ErC50 was calculated to be > 100 mg test item/L for frond number and dry weight, respectively.
The 7-day NOEyC and the LOEyC were determined to be 3.2 and 10 mg test item/L for frond number and dry weight, respectively.
The 7-day NOErC and the LOErC were determined to be 3.2 and 10 mg test item/L for frond number and dry weight, respectively.
No analytical dose verification was performed.
Executive summary:

Title:     Toxicity to the Aquatic Plant Lemna gibba in a Static Growth Inhibition Test

Purpose:             The purpose of this study was to determine the inhibitory effect of the test item on the growth of the freshwater aquatic plant Lemna gibba.

For this purpose, cultures of Lemna gibba were exposed in a static test to various concentrations under defined conditions. The inhibition of growth in relation to control cultures was determined over a test period of 7 days.

The test method of application and the test system are recommended by the test guidelines and Lemna gibba is the recommended test species.

No analytical dose verification was performed since the test substance (type UVCB) cannot be analysed by standard methods in such low test concentrations used in this study.

Guidelines/Recommendations:     Commission Regulation (EC) No 761/2009, Annex, Part C, C.26.: "Lemna sp. Growth Inhibition Test", Official Journal of the European Union (EN), dated August 24, 2009

     OECD Guideline 221: "Lemna sp. Growth Inhibition Test", adopted March 23, 2006

  OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", December 15, 2000

               

Material and Methods  

Test Item:          batch no.: Op 01/15; type: UVCB substance, according to certificate of analysis

Test Species:     Lemna gibba G 3

Test Design:     This study encompassed 6 treatment groups (5 dose rates of the test item and a control) with three replicates per test concentration and control.

At test start 12 fronds were introduced in each replicate and incubated for 7 days under static conditions. The frond numbers were determined on day 2, 5 and 7. The dry weight of each replicate was determined at test termination.

Endpoints:         Yield and growth rate based on frond number and dry weight.

Test Concentrations:     100, 32, 10, 3.2 and 1.0 mg test item/L and a control.

Test Conditions:              Water temperature: 22.2 to 24.9 °C; pH values at test start 7.4 to 7.6, at the end of the test 8.4 to 8.5; continuous illumination; mean light intensity: 8050 lux (7130 to 8670 lux).

Conclusion:        The influence of the test item on the growth of the freshwater plant Lemna gibba was assessed in a static concentration-response test.

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

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

The 7-day NOEyC and the LOEyC were determined to be 3.2 and 10 mg test item/L for frond number and dry weight, respectively.

The 7-day NOErC and the LOErC were determined to be 3.2 and 10 mg test item/L for frond number and dry weight, respectively.

No analytical dose verification was performed.

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