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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report date:
2020

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Ethanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with N,N-dimethylcyclohexanamine
EC Number:
948-066-8
Molecular formula:
C16H36NO6P C24H52NO8P C24H53NO11P2 C32H68NO10P C8H17N H3O4P
IUPAC Name:
Ethanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with N,N-dimethylcyclohexanamine
Test material form:
liquid: viscous
Specific details on test material used for the study:
Batch/Lot number: 0002293298
Appearance: High-viscous, colourless to yellowish liquid
Purity: Considered as 100%
Expiry date: 22 November 2019
Storage conditions: Controlled room temperature (15-25°C, ≤70% relative humidity)
Safety precautions: Routine safety precautions (gloves, goggles, face mask, lab coat) for unknown materials will be applied to assure personnel health and safety.

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
The concentration of the test item was measured at the test concentration at the start and at the end of each renewal period. Sample from the control was taken for analysis at least at the beginning of each renewal period.

Test solutions

Vehicle:
no
Details on test solutions:

A stock solution with a nominal concentration of 20 mg/L was prepared with direct addition of the test item, mixed into the test medium (OECD medium) using ultrasonic bath (approximately 3 minutes). The test solutions were prepared by appropriate diluting of this stock solution just before treatment (see Table 2 below).

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Source: Szent István University, 2100 Gödöllő, Páter Károly u. 1. – Hungary
Breeding: The Daphnia are bred in Ecotoxicological Laboratory of Citoxlab Hungary Ltd.
Justification of species: Daphnia magna is the standard species of the acute immobilisation test.
Previous generation: Daphnia of the previous generation should be reared with conditions similarly to the test conditions as possible. Test daphnia from stage with high reproduction activity (normally 2-4 weeks old) are preferred.
Number of animals: There will be 20 animals for each group (per concentration and untreated control) divided into 4 replicates of 5 animals.
Age of animals: Less than 24 hours old at the beginning of the test. The first breeding of the previous generation should not be used.
Acclimatisation: To avoid the necessity of adaptation prior to the test, the water used in the test will be similar to the culture conditions (water temperature, background colour etc.).
Holding water: Same water is used as test water (see section 2.3.).
Food and feeding: The Daphnia are fed with concentrated algal suspension of Pseudokirchneriella subcapitata at least three times per week during the holding.

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
No post exposure observation

Test conditions

Hardness:
The reconstituted water (ISO medium) had a total hardness of 249 mg/L (as CaCO3).
Test temperature:
20.4 – 20.7°C
pH:
7.33 – 7.87.
Dissolved oxygen:
The dissolved oxygen concentration was measured in one test vessel per group at the start and at the end of the test and was in the range of 8.1 – 8.5 mg/L.
Nominal and measured concentrations:
Because toxic response was not observed during the preliminary concentration range-finding test, a Limit Test was carried out using only one test concentration (100.0 mg/L) and one control group in a semi-static system.
The test concentration was analytically determined at the beginning and at the end of each renewal periods. Measured test item concentration was 99.69 mg/L at the start and 102.87 mg/L at the end of the first renewal period, 99.97 mg/L at the start and 102.30 mg/L at the end of the second renewal period, consequently, the corresponding measured geometric mean test item concentration was 101.20 mg/L.
As the measured concentrations deviated not more than 20% from the loading rate, the biological results are based on the nominal loading rate.
Details on test conditions:
Light:

The test was carried out in 16-hour light and 8-hour dark cycle.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
LOEC
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Results with reference substance (positive control):
The experimental period of the last study (Citoxlab Study Code: 19/009-023DA) with reference item Potassium dichromate (batch no.: A0345704) was 23 - 24 January 2019.
The observed 24h EC50 value for the reference material was 0.71 mg/L, (95 % confidence limits: 0.66 – 0.75 mg/L) in that study.

Any other information on results incl. tables

Table 2:Number and percentage of immobilised animals

Test Group

Number of treated animals

Number of immobilised animals

24 hours

48 hours

number

percent

number

percent

Control

20

0

0

0

0

100.0 mg/L (nominal)

20

0

0

0

0

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Acute toxicity of Ethanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with N,N-dimethylcyclohexanamine was assessed with Acute immobilisation test on Daphnia magna, over an exposure period of 48 hours in a semi-static system.

All validity criteria were met during this study.

Under the conditions of this Daphnia magna acute immobilisation study the observed endpoints for the effect of Ethanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with
N,N-dimethylcyclohexanamine were the followings:

The 24h and 48h EC50 value: > 100.0 mg/L (nominal)
The 48h EC100 value: > 100.0 mg/L (nominal)
The 48h No-Observed Effect Concentration (NOEC): 100.0 mg/L (nominal)
The 48h Lowest Observed Effect Concentration (LOEC): > 100.0 mg/L (nominal)
Executive summary:

Acute toxicity ofEthanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with N,N-dimethylcyclohexanamineonDaphniamagnawas assessed in an Acute immobilisation test, over an exposure period of 48 hours in a semi-static test system.

Because no toxic response was observed during the preliminary range-finding test, a Limit Test was carried outusing onlyone test concentration (100.0 mg/L)and one control groupin the definitive test.

 

The test concentrationwasanalytically determined at the beginning and at the end of each renewal periods. Measured test item concentration was 99.69 mg/L at the start and 102.87 mg/L at the end of the first renewal period, 99.97 mg/L at the start and 102.30 mg/L at the end of the second renewal period, consequently, the corresponding measured geometric mean test item concentration was 101.20 mg/L.

As the measured concentration deviated not more than 20% from the nominal concentration, biological results are based on the test item nominal concentration.

Twenty animals, divided into four groups (in individual glass beakers) of five animals each were used at the test concentration and for the control.

All validity criteria were met during this study.

 

Under the conditions of thisDaphnia magnaacute immobilisation study the observed endpoints for the effect ofEthanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with
N,N-dimethylcyclohexanaminewere the followings:

 

The 24h and 48h EC50value:       >100.0 mg/L (nominal)

The 48h EC100value:                        >100.0 mg/L (nominal)

The 48h No-Observed Effect Concentration (NOEC):         100.0 mg/L (nominal)

The 48h Lowest Observed Effect Concentration (LOEC):       >100.0 mg/L (nominal)