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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 May 2015 15 May 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
OGYI/38593-5/2012
Analytical monitoring:
yes
Details on sampling:
Measurement of the NPOC content of the samples:
1 M Hydrochloric acid: 2%
sparge time: 90 sec
injection volume: 50 μL
Vehicle:
not specified
Details on test solutions:
Because the test item was considered poorly soluble in water, a water soluble fraction (WSF) was prepared in accordance with the OECD guidance document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, OECD No. 23. A test item stock solution (nominal loading rate of 100 mg/L) was prepared by dispersing/dissolving the amount of test item into the test medium (ISO Medium). This solution was shaken for about 24 hours at approximately 30°C and then was equilibrated for about 24 hours at approximately 20°C. The non-dissolved test material was removed by filtration through a fine (0.22 μm) filter to give the 100 % saturated solution. The test solutions were prepared by appropriate dilution of this stock solution just before introduction of the Daphnia (prior to treatments).
The concentrations in the definitive test were based on the results of the preliminary range-finding test. The following nominal concentrations were tested 6.25, 12.5, 25, 50, and 100 % saturated solution based on a loading rate of 100 mg/L.
An analytical measurement of non-purgeable organic carbon was performed in acidified samples after purging with air.
Untreated Control: The dilution water (ISO-medium) was used without the test item.
Test organisms (species):
Daphnia magna
Details on test organisms:
Species and strain: Daphnia magna
Source: National Institute of Public Health, 1097 Budapest Gyáli u. 2-6. Hungary
Justification of strain: Daphnia magna is the standard species of the acute immobilisation test.
Number of animals: There were 20 animals in test and control groups respectively, divided into 4 replicates (5 animals/ replicate).
Age of the animals: They were less than 24 h old at the beginning of the test.
Acclimatization: There was no acclimatization because the water used was similar to the culture water.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
None specified
Hardness:
The reconstituted water (ISO medium) had a total hardness of 248 mg/L (as CaCO3).
Test temperature:
The water temperature was measured at the start and the end of each renewal period in each test vessels. The test temperature was in the range of
20.2 – 20.4°C measured in the test vessels. The additionally measured temperature in the climate chamber was between 19.5 and 20.6°C.
pH:
The pH of the test solution was not adjusted and not varied by more than 1.5 units in any one test. The pH was measured at the start and at the end of each renewal period in each test vessel and was in the range of 6.40 – 7.06.
Dissolved oxygen:
The dissolved oxygen concentration was measured in each test vessel at the start and at the end of each renewal period and was in the range of 8.1 – 8.9 mg/L.
Salinity:
Not applicable.
Nominal and measured concentrations:
Nominal concentrations were tested 6.25, 12.5, 25, 50, and 100 % saturated solution. The non-purgeable carbon content at a nominal loading rate of 100 mg/L was 36.02 mg/L
Details on test conditions:
The test was carried out in 16-hour light and 8-hour dark cycles. The immobility or mortality of the Daphnia was determined by visual observation 24 and 48 hours after the start of the test. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker are considered to be immobile.
The number of immobilized animals and the percentage of immobility were determined at 24 and 48 hours.
The water temperature, the oxygen concentrations and pH of the controls and the test solution were measured at the beginning and at the end of the renewal periods.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
NOELR
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
There were no immobilized animals in the control group and the dissolved oxygen concentration at the end of the test in control and test vessels was more than 3 mg/L. All validity criteria were within acceptable limits and therefore the study can be considered as valid.
The number of immobilised animals and the percentage of immobility were determined at the 24th and 48th hour. One immobilized animal (out of 20 in the whole group) was observed at a loading rate of 50 mg/L; this was considered to be due to natural causes rather than a toxic effect.
Results with reference substance (positive control):
For the evaluation of the quality of the Daphnia clone and the experimental conditions, Potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.
The date of the last study (Study Code: 15/081-023DA) with reference item Potassium dichromate (batch no.: A0345704) is: 03 - 04 March 2015.
The 24h EC50:1.05 mg/L, (95 % confidence limits: 0.89 – 1.23 mg/L)
Reported statistics and error estimates:
No statistical analysis was performed because of the lack of toxic effects.
The EL50 and NOELR values were determined directly from the raw data.

Table: Results of the Preliminary range-Finding Test

Nominal concentrations

[% saturated solution]

Untreated control

0.1

1

10

100

Number of treated / immobilized animals

10 / 0

10 / 0

10 / 0

10 / 2

10 / 2

Table: Number and percentage of immobilised animals in the definitive test

Nominal

Concentration

Number of treated animals

Immobilised animals

24 hours

48 hours

[% saturated solution]

number

[%]

number

[%]

Control

20

0

0

0

0

6.25

20

0

0

0

0

12.5

20

0

0

0

0

25

20

0

0

0

0

50

20

0

0

1

5

100

20

0

0

0

0

 

Immobilization of the test animals

Nominal Concentration [% saturated solution]

Replicate

 

Number of treated animals

 

Number of immobilised animals

24 h

48 h

Control

1

5

0

0

2

5

0

0

3

5

0

0

4

5

0

0

6.25

1

5

0

0

2

5

0

0

3

5

0

0

4

5

0

0

12.5

1

5

0

0

2

5

0

0

3

5

0

0

4

5

0

0

25

1

5

0

0

2

5

0

0

3

5

0

0

4

5

0

0

50

1

5

0

0

2

5

0

0

3

5

0

1

4

5

0

0

100

1

5

0

0

2

5

0

0

3

5

0

0

4

5

0

0

 

Validity criteria fulfilled:
yes
Conclusions:
The test item CUCURBIT[8]URIL had no toxic effect at a nominal loading rate of 100 mg/L [water-accommodated fraction (WAF); 36.02 mg/L measured non-purgeable organic carbon content]; the EL50 was >100 mg/L loading rate (WAF) and the NOELR was at 100 mg/L loading rate (WAF)
Executive summary:

The objective of the study was to determine the effect of CUCURBIT[8]URIL hydrochloride hydrate on Daphnia magna. Young Daphnia were exposed in a semi-static test to aqueous test medium containing the test item at different concentrations. The percentage of Daphnia being no longer capable of swimming at the end of the test or being dead was recorded for each test concentration. The GLP study was performed in accordance with OECD Guidelines for Testing of Chemicals, No.:202 (Adopted: 13 April 2004), the EPA Health Effects Test Guidelines, OCSPP 850.1010 and Commission Regulation (EC) No 761/2009 of 23 July 2009 amending, for the purpose of its adaptation to technical progress, Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACh), Annex IV Part C, C.2 (published in the Official Journal of the European Union L 220 of 24 August 2009).

The test item CUCURBIT[8]URIL had no toxic effect at a nominal loading rate of 100 mg/L [water-accommodated fraction (WAF); 36.02 mg/L measured non-purgeable organic carbon  content]; the EL50 was >100 mg/L loading rate (WAF) and the NOELR was at 100 mg/L loading rate (WAF).

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The source and target substances are structurally very similar with both consisting of the same monomeric units forming macrocylic compounds containing 6, 7 or 8 units (target substance) or 8 units (source substance). The functional groups are therefore identical. Source and target substance have similar physico-chemical properties and available toxicity data have revealed very similar toxicity profiles. Although not all starting materials for the synthesis are identical, the breakdown products - as far as known - do not differ. Both source and target form host guest complexes by a common mechanism of action. Therefore, read-across is justified.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The source substance is the hydrochloride hydrate of one of the constituents of the target substance. Neither source nor target substance contain toxicologically relevant impurities in relevant amounts.
3. ANALOGUE APPROACH JUSTIFICATION
All experimental data available to date are very similar for source and target substance (see data matrix) and confirm the analogue approach.
4. DATA MATRIX
see attachment
Reason / purpose:
read-across source
Key result
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 38.02 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat. (dissolved fraction)

Description of key information

Key values were read-across from an experimental study with an analogue substance which was performed in accordance with OECD Guideline 202, EU Method C2 and US EPA Procedure OCSPP 850.1010.

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
100 mg/L

Additional information

Under the conditions of this Daphnia magna acute immobilisation study the calculated and observed endpoints for the effect of CUCURBIT[8]URIL hydrochloride hydrate were the followings:

The 24h EC50 value: > 100 % saturated solution (nominal)

The 48h EC50 value: > 100 % saturated solution (nominal)

The 48h EC100 value: > 100 % saturated solution (nominal)

The 48h No-Observed Effect Concentration (NOEC):

> 100 % saturated solution (nominal).

The 48h Lowest Observed Effect Concentration (LOEC):

> 100 % saturated solution (nominal).

Based on the results of this study, the test item CUCURBIT[8]URIL had no toxic effect at a 100 mg/L

loading rate [36.02 mg/L measured NPOC (Non-Purgeable Organic Carbon) content, corresponding to 100 mg/L CUCURBIT[8]URIL]]