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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

Link to relevant study record(s)

Referenceopen allclose all

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:
other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000.
Qualifier:
according to guideline
Guideline:
ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
Preparation of solutions
Stock and spiking solutions
Stock solutions of the test substance were prepared in acetonitrile at concentrations of 2000 and 3001 mg/l.
Spiking solutions were made up from a stock solution and/or dilutions of this solution. The solvent of the spiking solutions was acetonitrile.
Calibration solutions
Calibration solutions in the concentration range of 0.04 – 10 mg/l were prepared from two stock solutions. The end solution of the calibration solutions was 50/50 (v/v) acetonitrile/water.
Procedural recovery samples
2 ml blank medium was spiked with the test substance at a target concentration of 0.1 or 100 mg/l.
The accuracy samples were treated similarly as the test samples

Blank procedural recovery samples were prepared and treated similarly to the test samples.

Sample injections
Calibration solutions were injected in duplicate. Test samples and procedural recovery samples were analysed by single injection.
The test samples were stored in the freezer (≤ -15°C).
On the day of analysis, the test samples were defrosted at room temperature. The samples were diluted in a 1:1 (v:v) ratio with acetonitrile and analysed. If necessary, the samples were further diluted with 50/50 (v/v) acetonintrile/ISO-medium to obtain concentrations within the calibration range.
Vehicle:
no
Details on test solutions:
The standard test procedures required generation of test solutions, which should contain completely dissolved test substance concentrations or stable and homogeneous mixtures or dispersions. The testing of concentrations that disturb the test system should be prevented (e.g. film of the test substance on the water surface). No correction was made for the purity/composition of the test substance.
The batch of ANOX® 1315 tested was a clear yellow viscous liquid with a purity of 93.5% by GC and not completely soluble in test medium at the loading rates initially prepared.
Preparation of test solutions started with individual loading rates of 1.0, 10 and 100 mg/l. A two-day period of magnetic stirring was applied and was followed by a one-hour settlement period. Thereafter, the aqueous Water Accommodated Fractions (WAFs) were siphoned off and used as test solutions. The final test solutions were all clear and colourless.
Test organisms (species):
Daphnia magna
Details on test organisms:
Test system
Species Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions.
Source In-house laboratory culture with a known history.
Reason for selection This system has been selected as an internationally accepted invertebrate species.
Validity of batch Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.
Characteristics For the test selection of young daphnids with an age of < 24 hours, from parental daphnids of more than two weeks old.

Breeding
Start of each batch With newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel.
Maximum age of the cultures 4 weeks
Renewal of the cultures After 7 days of cultivation half of the medium twice a week.
Temperature of medium 18-22°C
Feeding Daily, a suspension of fresh water algae.
Medium M7, as prescribed by Dr. Elendt-Schneider
(Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33).
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
180 mg/l CaCO3
pH:
7.7 ± 0.3
Dissolved oxygen:
≥3 mg/l
Salinity:
Not specified
Conductivity:
Not specified
Nominal and measured concentrations:
Limit test: Nominal concentrations of 1.0, 10 and 100 mg/l
Main test: Nominal concentration of 1.0 mg/l
Details on test conditions:
Test concentrations
ANOX® 1315: WAF prepared at a loading rate of 1.0 mg/l.
Controls: Test medium without test substance or other additives

Test procedure and conditions
Test duration: 48 hours
Test type: Static
Test vessels: 100 ml, all-glass
Medium: Adjusted ISO medium
Number of daphnids: 20 per group
Loading: 5 per vessel containing 80 ml of test solution
Light: 16 hours photoperiod daily
Feeding: No feeding
Aeration: No aeration of the test solutions.
Introduction of daphnids: Within 45 minutes after preparation of the test solutions.
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 0.17 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
Combined limit/range-finding test
No immobility was observed in the control and the lowest concentration during the exposure.
Effects observed in the two highest WAFs were caused by undissolved test material as a floating layer was observed at the surface of the test solutions and daphnids were stuck with each other.
This floating layer was not present in vessels at the start of the test but was observed after 2½ hour.
Samples taken from WAFs prepared at 1.0 and 100 mg/l were analysed. The initially measured concentrations were 0.28 and 14 mg/l, respectively. At the end of the test measured concentrations were at the level of 80-137% of initial.
Because the solubility of the test substance in water was 0.33 mg/l it was decided, in consultation with the sponsor, to perform the final test as a limit with a WAF prepared at a loading rate of 1.0 mg/l to avoid effects caused by undissolved test material.

Limit test
The concentration measured in the WAF prepared at 1.0 mg/l was 0.17 mg/l at the start of the test and remained stable during the exposure period (87% of initial at the end of the test). Therefore, the effect parameters were expressed in terms of initially measured concentrations.

Immobility
The responses recorded in this test allowed for reliable determination of an EC50. The responses recorded at the limit concentration were in agreement with the results of the combined limit/range-finding test. No significant immobilisation (i.e. >10%) was observed during this test.

Experimental conditions
These test conditions remained within the limits prescribed by the protocol (pH: 6.0-8.5, not varying by more than 1.5 units; oxygen: ≥3 mg/l at the end of the test).
The temperature of the test medium was 20°C at the start of the test. The temperature continuously measured in a temperature control vessel varied between 21 and 22°C during the test, and complied with the requirements as laid down in the protocol (18-22°C, constant within 2°C).
Results with reference substance (positive control):
The actual responses in this reference test with K2Cr2O7 are within the ranges of the expected responses at the different concentrations, i.e. the 48h-EC50 was between 0.3 and 1.0 mg/l. Hence, the sensitivity of the daphnia was within the range determined with the historical data.
The 24h-EC50 was 0.70 mg/l with a 95% confidence interval between 0.62 and 0.80 mg/l.
The 48h-EC50 was 0.41 mg/l with a 95% confidence interval between 0.36 and 0.47 mg/l.

Number of introduced daphnids and incidence of immobility in the combined limit/range-finding test

























































































































































Time (h)


 



Replicate


 



ANOX® 1315, WAF prepared at mg/l


 



Control


 



1.0



10



100



0



A



5



5



5



5



 



B



5



5



5



5



 



C



5



 



 



5



 



D



5



 



 



5



 



Total introduced


 



20



10



10



20



24



A



0 (1)


 



0



0 (5)



3 (4)



 



B



0



0 (2)



0 (4)



4 (5)



 



C



0



 



 



2 (5)



 



D



0



 



 



1 (3)



 



Total immobilised


 



0



0



0



10



 



Effect %



0



0



0



50



48



A



0



0



4



5 (3)



 



B



0



0 (1)



3 (1)



5 (1)



 



C



0



 



 



4



 



D



0



 



 



5



 



Total immobilised


 



0



0



7



19



 



Effect %



0



0



70



95



( ) between brackets: number of daphnia observed trapped at the surface of the test solutions. These organisms were reimmersed into the respective solutions before recording of mobility.


 


Number of introduced daphnids and incidence of immobility in the limit test





















































































































Time (h)


 



Replicate


 



ANOX® 1315, WAF prepared at mg/l


 



Control


 



1.0



0



A



5



5



 



B



5



5



 



C



5



5



 



D



5



5



 



Total introduced


 



20



20



24



A



0


 



0 (5)



 



B



0



0 (2)



 



C



0



0



 



D



0



0



 



Total immobilised


 



0



0



 



Effect %



0



0



48



A



0



0



 



B



1



0



 



C



0



0 (1)



 



D



0



0



 



Total immobilised


 



1



0



 



Effect %



5



0



( ) between brackets: number of daphnia observed trapped at the surface of the test solutions. These organisms were reimmersed into the respective solutions before recording of mobility.


 


Determination of effect concentrations














Parameter


 



Initial concentration ANOX® 1315


(mg/l)



24 and 48h-EC50


 



0.17



 


pH and oxygen concentrations during the limit test































Initial concentration


ANOX® 1315


(mg/l)



Start (t=0 h)


 



End (t=48 h)


 



pH



O2



pH



O2



control


 



7.9


 



8.7


 



8.1


 



9.3


 



0.17


 



8.0


 



8.8


 



8.1


 



9.2


 



 


Acute immobilization of daphnia after 24 and 48 hours in the reference test with potassium dichromate:










































































Concentration


K2Cr2O7


(mg/l)


 



Number


Exposed



% immobile


 



Expected response (%)


After 48 hours



24h


 



48h


 



Minimal


 



Maximal


 



control


 



20


 



0


 



0


 



0



102


 



0.10


 



20


 



0


 



0


 



0



10



0.18


 



20


 



0


 



0


 



0



10



0.32


 



20


 



0


 



15



0



30



0.56


 



20


 



15



90



0



100



1.0


 



20


 



95



100



40



100



1.8


 



20


 



100



100



100



100



1 Based on historical data of the previous years (n>60).


2 A maximum response of 10% does not invalidate the results of the test.

Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of the present study the 48h-EC50 was beyond the range tested, i.e. exceeded an initially measured concentration of 0.17 mg/l being considered the maximum soluble in the medium.
Executive summary:

Acute Toxicity Study in Daphnia magna with ANOX® 1315.



The study procedures described in this report were based on the OECD guideline No. 202, 2004. In addition, the procedures were designed to meet the test methods of the Commission Regulation (EC) No 440/2008, Part C.2, 2008, the ISO International Standard 6341, 1996 and the OECD series on testing and assessment number 23, 2000.


The batch of ANOX® 1315 tested was a clear yellow viscous liquid with a purity of 93.5% by GC and not completely soluble in test medium at the loading rates initially prepared.


A limit test was performed based on the results of a preceding combined limit/range-finding test. A Water Accommodated Fraction (WAF) was prepared at a loading rate of 1.0 mg/l. A two-day period of magnetic stirring was applied and was followed by a one-hour settlement period. Thereafter, the aqueous WAF was siphoned off and used as test solution. The final test solution was clear and colourless.


Twenty daphnids per group (5 per replicate, quadruplicate) were exposed to an untreated control and the limit concentration, i.e. WAF prepared at a loading rate of 1.0 mg/l. The total exposure period was 48 hours and samples for analytical confirmation of exposure concentrations were taken at the start and at the end of the test.


The actual concentration measured in the WAF prepared at 1.0 mg/l was 0.17 mg/l at the start of the test and remained stable during the exposure period (87% of initial at the end of the test). Therefore, the effect parameters were expressed in terms of initially measured concentrations.


The study met the acceptability criteria prescribed by the protocol and was considered valid.


The 48h-EC50 was beyond the range tested, i.e. exceeded an initially measured concentration of 0.17 mg/l being considered the maximum soluble in the medium.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
23 September 1992 - 25 September 1992
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)
GLP compliance:
yes
Analytical monitoring:
not specified
Test organisms (species):
Daphnia magna
Details on test organisms:
Species: Daphnia Magna
Justification for the section of the test sysfem: Daphnia Magna was chosen as a Crustacean species since it is widely accepted by Health Authorities as an experimental model with documented susceptibility to a wide range of toxic substances.
Number: 20 animals/concentration subdivided into batches of 5 animals each
Age at the beginning of test: not more than 24 hours.
Supplier: laboratory breeding
Housing: the animals were bred in glass containers. To avoid the necessity of adaptation prior to the test, the water used in the test was similar to the culture water.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
Not specified
Test temperature:
20
pH:
7.9
Dissolved oxygen:
3.5 mg/l
Salinity:
Not specified
Conductivity:
Not specified
Nominal and measured concentrations:
Nominal concentration of 0.33 mg/l
Details on test conditions:
Test system
Light: 12-16 hours of photoperiod daily.
During the test the animals were kept in the dark.
Feeding: the animals were fed with (laboratory grown) green algae (Selenastrum capricornutum) three times a week.
During the treatment period the animals were not fed.
Contaminants that might interfere with the objectives of the study were not expected to be present in feed or water.
Housing during the treatment: 20 animals/stoppered glass beaker containing 40 milliliters of treatment solution.
Vessel identification gave experimental number and group number.

Test description
Experimental design: on request of the Sponsor the test
article was tested at the nominal concentration of 0.33 mg/l that is the maximum solubility of the test article in water.
One group of 20 animals was treated only with water (control group).
Treatment frequency: single
Treatment period: 48 hours
Observation period: 48 hours
Observation of immobilization at 24 and 48 hours. Daphniae were considered immobile if not able to swim within 15 seconds after gentle agitation of the test container.
Parameter checking: at 0 and 48 hours of treatment the following parameters were checked in the treatment solutions:
- dissolved oxygen by means of the oxymeter YIS model 54 RL
- pH by means of the pH meter Radiometer PHM SO standard
- temperature by means of a mercuric thermometer
EC50 at 24 and 48 hours and its statistical limits: were not calculable.
Loading: 1 animal/2 ml

Formulation of test solution
Preparation: a solution was prepared by dissolving a weighed amount of test article directly in the treatment water in order to obtain the requested concentration.
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 0.33 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
> 0.33 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
Immobilization
No animals in the treated group nor in the control group were immobilized.

Parameter checking
The values of dissolved oxygen, pH and temperature always remained between the acceptable limits.

EC50 and its statistical limits
The values of EC50 at 24 and 48 hours were not calculable
Validity criteria fulfilled:
yes
Conclusions:
Experimental data from an acute immobilization test in which Daphnia magna were treated with the test article ANOX BF at the nominal concentration of 0.33 mg/l (maximum solubility in water).
The animals were treated and observed for 48 hours.
No animals in the treated group nor in the control group were immobilized.
The EC50 at 24 and 48 hours were not calculable.
Executive summary:

The purpose of rhe study was to evaluate the ecotoxicity of the test article ANOX BF to determine the EC50 at 24 and 48 hours, its statistical limits and slope, if possible.

Experimental data from an acute immobilization test in which Daphnia magna were treated with the test article ANOX BF at the nominal concentration of 0.33 mg/l (maximum solubility in water).
The animals were treated and observed for 48 hours.
No animals in the treated group nor in the control group were immobilized.
The EC50 at 24 and 48 hours were not calculable.

Description of key information

ANOX BF Acute immobilization study in Daphnia Magna

The purpose of the study was to evaluate the ecotoxicity of the test article ANOX BF to determine the EC50 at 24 and 48 hours, its statistical limits and slope, if possible.

Experimental data from an acute immobilization test in which Daphnia magna were treated with the test article ANOX BF at the nominal concentration of 0.33 mg/l (maximum solubility in water).
The animals were treated and observed for 48 hours.
No animals in the treated group nor in the control group were immobilized.
The EC50 at 24 and 48 hours were not calculable.

 

Acute Toxicity Study in Daphnia magna with ANOX® 1315

Under the conditions of the present study the 48h-EC50 was beyond the range tested, i.e. exceeded an initially measured concentration of 0.17 mg/l being considered the maximum soluble in the medium.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
100 mg/L

Additional information

Acute Toxicity Study in Daphnia magna with ANOX® 1315.

The study procedures described in this report were based on the OECD guideline No. 202, 2004. In addition, the procedures were designed to meet the test methods of the Commission Regulation (EC) No 440/2008, Part C.2, 2008, the ISO International Standard 6341, 1996 and the OECD series on testing and assessment number 23, 2000.
The batch of ANOX® 1315 tested was a clear yellow viscous liquid with a purity of 93.5% by GC and not completely soluble in test medium at the loading rates initially prepared.
A limit test was performed based on the results of a preceding combined limit/range-finding test. A Water Accommodated Fraction (WAF) was prepared at a loading rate of 1.0 mg/l. A two-day period of magnetic stirring was applied and was followed by a one-hour settlement period. Thereafter, the aqueous WAF was siphoned off and used as test solution. The final test solution was clear and colourless.
Twenty daphnids per group (5 per replicate, quadruplicate) were exposed to an untreated control and the limit concentration, i.e. WAF prepared at a loading rate of 1.0 mg/l. The total exposure period was 48 hours and samples for analytical confirmation of exposure concentrations were taken at the start and at the end of the test.
The actual concentration measured in the WAF prepared at 1.0 mg/l was 0.17 mg/l at the start of the test and remained stable during the exposure period (87% of initial at the end of the test). Therefore, the effect parameters were expressed in terms of initially measured concentrations.
The study met the acceptability criteria prescribed by the protocol and was considered valid.
The 48h-EC50 was beyond the range tested, i.e. exceeded an initially measured concentration of 0.17 mg/l being considered the maximum soluble in the medium.