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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
12-05-2017 to 23-08-2017
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Guideline study performed under GLP. All relevant validity criteria were met. The source and target substances are related by virtue of the source substance being a ‘common breakdown product’ of the target substance. The source and target demonstrate ‘chemical similarity’ : since the source is the structurally-similar degradation product (i.e. similarity through (bio) transformation) from the target. The target substance hydrolytically degrades in water in a matter of seconds/minutes at all relevant pH 4, 7 and 9 forming the source substance. See related endpoint information in IUCLID: section 5.1.2: hydrolysis.
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included in attachment to IUCLID section 13.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The read-across is based on the hypothesis that the source and target substances are related by virtue of the source substance being a ‘common breakdown product’ of the target substance. The source and target demonstrate ‘chemical similarity’ : since the source is the structurally-similar degradation product (i.e. similarity through (bio) transformation) from the target. The target substance hydrolytically degrades in water in a matter of seconds/minutes at all relevant pH 4, 7 and 9 forming the source substance. See related endpoint information in IUCLID: section 5.1.2: hydrolysis.
Further information is included in attachment to IUCLID section 13.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The source and target chemicals have comparable chemical similarity. Further information is included in attachment to IUCLID section 13

3. ANALOGUE APPROACH JUSTIFICATION
The source substance is a the ‘common breakdown product’ of the target substance. The source and target demonstrate ‘chemical similarity’ : since the source is the structurally-similar degradation product (i.e. similarity through (bio) transformation) from the target. The target substance hydrolytically degrades in water in a matter of seconds/minutes at all relevant pH 4, 7 and 9 forming the source substance. As a consequence they should be considered chemically similar substances due to routes of common (a)biotic degradation and/or metabolism and common or similar degradants. See related endpoint information in IUCLID: section 5.1.2: hydrolysis. Further information is included in attachment to IUCLID section 13

4. DATA MATRIX
Further information is included in attachment to IUCLID section 13.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across: supporting information
Remarks:
Reporting format for the analogue approach
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
2004
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
All concentration levels and the control were analytically verified via HPLC-DAD at the start (0 hours) and at the end of the exposure (48 hours) in all concentration levels and in the control. At the start of the exposure (0 hours), sampling was carried out after preparation of the test item concentrations. At the end of the exposure (48 hours), samples of the old media were taken from test vessels.
Vehicle:
no
Details on test solutions:
The stock solution (100 mg/L of the test item were weighed out) was prepared with dilution water (see Table 1) one day prior to the start of the exposure (at -24hours). The stock solution was stirred with a magnetic stirrer approximately 1100 rpm for 24 hours at room temperature. 5 test item concentrations in a geometric series with a separation factor of 2, prepared by dilution of the stock solution of 100 mg/L with dilution water (see Table 1), were tested as follows: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
The test concentrations were selected based on the results of a non-GLP preliminary range finding test.
Control: Dilution water without test item incubated under the same conditions as the test groups
Test organisms (species):
Daphnia magna
Details on test organisms:
Less than 24 hours old daphnids from a healthy stock were used for the study. Juvenile daphnids were removed from the culture vessels at the latest 24 hours before the start of the exposure and discarded. The juveniles born within the following period of max. 24 hours preceding the exposure were used for the test. No first brood progeny was used for the test. Acclimatization was not necessary, because the dilution water was equivalent to the culture medium.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Test temperature:
18 - 22 °C, constant within ± 1 °C
pH:
Control: 7.59
Test solutions: 7.20 to 7.54
Dissolved oxygen:
Control: 9.07 mg/L
Test solutions: 8.48 to 8.96 mg/L
Nominal and measured concentrations:
nominal : 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
measured: 6.56- 13.0- 25.3- 52.1- 104 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers, 50 mL
- Type (delete if not applicable): loosely covered with watch glasses
- Material, size, headspace, fill volume: Glass beakers (4 (10) x 7 (H) cm), 50 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: see above
- Conductivity: Dilution water at test start: 448 µS/cm
- Culture medium different from test medium: Elendt M4 acc. to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: At test start and test end

OTHER TEST CONDITIONS
- Adjustment of pH: yes in the highest concentration of 100 mg/L by addition of 1 M NaOH prior to start of the exposure.
- Photoperiod: 16:8 light : dark cycle
- Light intensity: max. 1500 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility was determined in all groups after 24 h and 48 h.

TEST CONCENTRATIONS
- Range finding study:
Immobilization Rates in the non GLP Preliminary Range Finding Test
(n = 20, divided into 2 replicates with 10 daphnids each)

Nominal
test item concentration [mg/L] / IMMOBILIZATION [%]
24 hours Replicates and 48 hours Replicates
1 / 2 / MV : 1 / 2 / MV
100 : 100 / 100 / 100 : 100 / 100 / 100 : % mortality after 24 hours
10 : 0 / 0 / 0 : 0 / 0 / 0
1 : 0 / 0 / 0 : 0 / 0 / 0
Control : 0 / 0 / 0 : 0 / 0 / 0
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
70.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: pH not adjusted
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: pH adjusted for neutralisation
Results with reference substance (positive control):
Test concentrations
1.00 – 2.00 – 4.00 mg/L
Current Study Valid Range
EC50 1.60 mg/L 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000); clone 5
0.6 - 2.1 mg/L, acc. to OECD 202 (2004); clone A
95% confidence limits 1.43- 1.79mg/L
Test duration: 04-05-2017 to 05-05-2017

Measured Concentrations and Percent of Nominal Concentration of the Test ltem: benzene-1,3,5-tricarboxylicacid during the DefinitiveTest


 









































































Samplingdate



2017-06-07


Start of the exposure, 0hours



2017-06-09


End of the exposure, 48hours



Start ofanalysis



2017-06-07



2017-06-09



 


Nominal


test itemconcentration [mg/L]



benzene-1,3,5-tricarboxylic acid



Meas. conc.rmCl/Ll



 


%



Meas. conc.[mC1/Ll



 


%



100*



107



107



104



104



100



106



106



105



105



50.0



51.8



104



52.1



104



25.0



26.0



104



25.3



101



12.5



13.2



105



13.0



104



6.25



6.57



105



6.56



105



Control



<LOQ



<LOQ



 Meas.conc.=measuredconcentrationofthetestitem,meanvalueof2injections,dilutionfactorstakenintoaccount


% = percent of the nominal concentration of the test item


LOQ = limit of quantification of the analytical method (0.300 mg/L of the test   item)


* =pH-neutralised


 


Absolute Numbers of immobile Daphnids after 24 and 48h of Exposure in the DefinitiveTest (n=20,divided into 4 replicates with 5daphnids each)












































































































































benzene-1,3,5-



NUMBER



OF IMMOBILEDAPHNIDS



ITOTAL NUMBEROF



DAPHNIDS



tricarboxylicacid



24h



 


48h



Nominal



Replicates



Replicates



testitem



 


1



 


2



 


3



 


4



 


MV



 


1



 


2



 


3



 


4



 


MV



concentration



rmg/LJ



100*



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



100



5/



5



5 /5



5 /5



5 /5



20/20



5 /5



5 /5



5 /5



5 /5



20/20



50.0



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



25.0



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



12.5



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



6.25



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



Control



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



* =pH-neutralised


 

Validity criteria fulfilled:
yes
Conclusions:
For the target substance: the 48 hour-EC50 (invertebrates) is considered to be 70.7 mg/L based on analytically confirmed nominal concentrations.
Executive summary:

The acute immobilisation toxicity of a source substance to Daphnia magna (STRAUS) was tested according to the principles of OECD TG 202 and EU Method C.2 in accordance with GLP. The study was conducted under static conditions over a period of 48 hours. Five test item concentrations in a geometric series with a separation factor of 2, prepared by dilution of the stock solution of 100.0 mg/L with dilution water, were tested as follows: 0 (control), 6.25, 12.5, 25.0, 50.0 and 100 mg/L. Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control. The concentrations of the test item were analytically verified via HPLC-DAD at the start (0 hours) and at the end of the exposure (48 hours) in all concentration levels and in the control. A further group of twenty daphnids (divided into 4 replicates with 5 daphnids each) were tested in the highest concentration of 100 mg/L by addition of 1 M NaOH prior to start of the exposure under the same test conditions as the test vessels as a 'pH control'. The measured concentrations of the test item were in the range of 104 to 107% of the nominal values at the start of the exposure (0 hours) and 101 to 105% of the nominal values at the end of the exposure (48 hours). All validity criteria were considered to be met. Based on analytically confirmed nominal concentrations of the test item the 48 hour-EC50 for Daphnia magna > 100 mg/L for adjusted pH and 70.7 mg/L for non-adjusted pH
For the target substance: the 48 hour-EC50 (invertebrates) is considered to be 70.7 mg/L based on analytically confirmed nominal concentrations.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12-05-2017 to 23-08-2017
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:
2004
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
All concentration levels and the control were analytically verified via HPLC-DAD at the start (0 hours) and at the end of the exposure (48 hours) in all concentration levels and in the control. At the start of the exposure (0 hours), sampling was carried out after preparation of the test item concentrations. At the end of the exposure (48 hours), samples of the old media were taken from test vessels.
Vehicle:
no
Details on test solutions:
The stock solution (100 mg/L of the test item were weighed out) was prepared with dilution water (see Table 1) one day prior to the start of the exposure (at -24hours). The stock solution was stirred with a magnetic stirrer approximately 1100 rpm for 24 hours at room temperature. 5 test item concentrations in a geometric series with a separation factor of 2, prepared by dilution of the stock solution of 100 mg/L with dilution water (see Table 1), were tested as follows: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
The test concentrations were selected based on the results of a non-GLP preliminary range finding test.
Control: Dilution water without test item incubated under the same conditions as the test groups
Test organisms (species):
Daphnia magna
Details on test organisms:
Less than 24 hours old daphnids from a healthy stock were used for the study. Juvenile daphnids were removed from the culture vessels at the latest 24 hours before the start of the exposure and discarded. The juveniles born within the following period of max. 24 hours preceding the exposure were used for the test. No first brood progeny was used for the test. Acclimatization was not necessary, because the dilution water was equivalent to the culture medium.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Test temperature:
18 - 22 °C, constant within ± 1 °C
pH:
Control: 7.59
Test solutions: 7.20 to 7.54
Dissolved oxygen:
Control: 9.07 mg/L
Test solutions: 8.48 to 8.96 mg/L
Nominal and measured concentrations:
nominal : 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
measured: 6.56- 13.0- 25.3- 52.1- 104 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers, 50 mL
- Type (delete if not applicable): loosely covered with watch glasses
- Material, size, headspace, fill volume: Glass beakers (4 (10) x 7 (H) cm), 50 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: see above
- Conductivity: Dilution water at test start: 448 µS/cm
- Culture medium different from test medium: Elendt M4 acc. to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: At test start and test end

OTHER TEST CONDITIONS
- Adjustment of pH: yes in the highest concentration of 100 mg/L by addition of 1 M NaOH prior to start of the exposure.
- Photoperiod: 16:8 light : dark cycle
- Light intensity: max. 1500 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility was determined in all groups after 24 h and 48 h.

TEST CONCENTRATIONS
- Range finding study:
Immobilization Rates in the non GLP Preliminary Range Finding Test
(n = 20, divided into 2 replicates with 10 daphnids each)

Nominal
test item concentration [mg/L] / IMMOBILIZATION [%]
24 hours Replicates and 48 hours Replicates
1 / 2 / MV : 1 / 2 / MV
100 : 100 / 100 / 100 : 100 / 100 / 100 : % mortality after 24 hours
10 : 0 / 0 / 0 : 0 / 0 / 0
1 : 0 / 0 / 0 : 0 / 0 / 0
Control : 0 / 0 / 0 : 0 / 0 / 0
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
70.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: pH not adjusted
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: pH adjusted for neutralisation
Results with reference substance (positive control):
Test concentrations
1.00 – 2.00 – 4.00 mg/L
Current Study Valid Range
EC50 1.60 mg/L 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000); clone 5
0.6 - 2.1 mg/L, acc. to OECD 202 (2004); clone A
95% confidence limits 1.43- 1.79mg/L
Test duration: 04-05-2017 to 05-05-2017

Measured Concentrations and Percent of Nominal Concentration of the Test ltem: benzene-1,3,5-tricarboxylicacid during the DefinitiveTest


 









































































Samplingdate



2017-06-07


Start of the exposure, 0hours



2017-06-09


End of the exposure, 48hours



Start ofanalysis



2017-06-07



2017-06-09



 


Nominal


test itemconcentration [mg/L]



benzene-1,3,5-tricarboxylic acid



Meas. conc.rmCl/Ll



 


%



Meas. conc.[mC1/Ll



 


%



100*



107



107



104



104



100



106



106



105



105



50.0



51.8



104



52.1



104



25.0



26.0



104



25.3



101



12.5



13.2



105



13.0



104



6.25



6.57



105



6.56



105



Control



<LOQ



<LOQ



 Meas.conc.=measuredconcentrationofthetestitem,meanvalueof2injections,dilutionfactorstakenintoaccount


% = percent of the nominal concentration of the test item


LOQ = limit of quantification of the analytical method (0.300 mg/L of the test   item)


* =pH-neutralised


 


Absolute Numbers of immobile Daphnids after 24 and 48h of Exposure in the DefinitiveTest (n=20,divided into 4 replicates with 5daphnids each)












































































































































benzene-1,3,5-



NUMBER



OF IMMOBILEDAPHNIDS



ITOTAL NUMBEROF



DAPHNIDS



tricarboxylicacid



24h



 


48h



Nominal



Replicates



Replicates



testitem



 


1



 


2



 


3



 


4



 


MV



 


1



 


2



 


3



 


4



 


MV



concentration



rmg/LJ



100*



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



100



5/



5



5 /5



5 /5



5 /5



20/20



5 /5



5 /5



5 /5



5 /5



20/20



50.0



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



25.0



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



12.5



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



6.25



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



Control



0/



5



0 /5



0 /5



0 /5



0/20



0 /5



0 /5



0 /5



0 /5



0/20



* =pH-neutralised


 

Validity criteria fulfilled:
yes
Conclusions:
In this study, based on analytically confirmed nominal concentrations of the test item the 48 hour-EC50 for Daphnia magna >100 mg/L for adjusted pH and 70.7 mg/L for non-adjusted pH
The validity criteria of the test guidelines were fulfilled.
Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS), the effects of the test item were determined according to OECD 202 (2004), which is equivalent to the Council Regulation (EC) No.440/2008 Method C.2 (2008). The study was conducted under static conditions over a period of 48 hours. Five test item concentrations in a geometric series with a separation factor of 2, prepared by dilution of the stock solution of 100.0 mg/L with dilution water, were tested as follows: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L.


Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control. The concentrations of the test item were analytically verified via HPLC-DAD at the start (0 hours) and at the end of the exposure (48 hours) in all concentration levels and in the control.


The measured concentrations of the test item were in the range of 104 to 107% of the nominal values at the start of the exposure (0 hours) and 101 to 105% of the nominal values at the end of the exposure (48 hours).


Based on analytically confirmed nominal concentrations of the test item the 48 hour-EC50 for Daphnia magna >100 mg/L for adjusted pH and 70.7 mg/L for non-adjusted pH

Description of key information

Short-term toxicity to aquatic invertebrates: target : benzene-1,3,5-tricarbonyl trichloride : 48h-EC50 (invertebrates) = 70.7 (C.I. -) mg/L based on analytically confirmed concentrations, 48-hour, freshwater, OECD TG 202, 2017
Read-Across: source data : benzene-1,3,5-tricarboxylic acid : OECD TG 201, 2017


Note: The source and target substances are related by virtue of the source substance being a ‘common breakdown product’ of the target substance. The target substance hydrolytically degrades in water in a matter of seconds/minutes at all relevant pH 4, 7 and 9 forming the source substance. The read-across hypothesis has been demonstrated experimentally.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Dose descriptor:
EC50
Effect concentration:
70.7 mg/L

Additional information

Key study : Read-Across: TARGET (benzene-1,3,5-tricarbonyl trichloride) from SOURCE (benzene-1,3,5-tricarboxylic acid) : OECD TG 201, 2017 : The acute immobilisation toxicity of a source substance to Daphnia magna (STRAUS) was tested according to the principles of OECD TG 202 and EU Method C.2 in accordance with GLP. The study was conducted under static conditions over a period of 48 hours. Five test item concentrations in a geometric series with a separation factor of 2, prepared by dilution of the stock solution of 100.0 mg/L with dilution water, were tested as follows: 0 (control), 6.25, 12.5, 25.0, 50.0 and 100 mg/L. Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control. The concentrations of the test item were analytically verified via HPLC-DAD at the start (0 hours) and at the end of the exposure (48 hours) in all concentration levels and in the control. A further group of twenty daphnids (divided into 4 replicates with 5 daphnids each) were tested in the highest concentration of 100 mg/L by addition of 1 M NaOH prior to start of the exposure under the same test conditions as the test vessels as a 'pH control'. The measured concentrations of the test item were in the range of 104 to 107% of the nominal values at the start of the exposure (0 hours) and 101 to 105% of the nominal values at the end of the exposure (48 hours). All validity criteria were considered to be met. Based on analytically confirmed nominal concentrations of the test item the 48 hour-EC50 for Daphnia magna > 100 mg/L for adjusted pH and 70.7 mg/L for non-adjusted pH
For the target substance: the 48 hour-EC50 (invertebrates) is considered to be 70.7 mg/L based on analytically confirmed nominal concentrations.


Note: The source (benzene-1,3,5-tricarboxylic acid) and target (benzene-1,3,5-tricarbonyl trichloride) substances are related by virtue of the source substance being a ‘common breakdown product’ of the target substance. The target substance hydrolytically degrades in water in a matter of seconds/minutes at all relevant pH 4, 7 and 9 forming the source substance. The read-across hypothesis has been demonstrated experimentally in other endpoints. Refer to 'Biodegradation in water: screening tests - AE 1' section for further information.