Reaction products of 2,3-epoxypropyl phenyl ether and 2,2'-iminodi(ethylamine)

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:

22 May 2018 - 08 May 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:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of 2,3-epoxypropyl phenyl ether and 2,2'-iminodi(ethylamine)

EC Number:

948-518-4

Molecular formula:

(C13H23N3O2 . C31H43N3O6)x

IUPAC Name:

Reaction products of 2,3-epoxypropyl phenyl ether and 2,2'-iminodi(ethylamine)

Test material form:

liquid

Details on test material:

PGE-DETA adduct

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Olin Corporation Batch: 17-09-502-04 QM 14K17-54
- Expiration date of the lot/batch: 30 October 2019
- Purity test date: Approx. 97% (including the oligomeric isomers)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Concentrations: 0, 1.0, 3.2, 10, 32, and 100 mg/L
Samples were taken from the control and each test group from the bulk test preparation at 0 hours and from the pooled replicates (R1 to R4) at 48 hours for quantitative analysis. All samples were stored frozen prior to analysis.

Test solutions

Vehicle:

no

Details on test solutions:

Nominal amounts of test item (5, 16, 50, 160 and 500 mg) were each separately added to the surface of 5 liters of test water to give the 1.0, 3.2, 10, 32 and 100 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1-Hour. Visual observations made on the WAFs indicated that a significant amount of dispersed test item was present in the water column and hence it was considered justifiable to remove the WAFs by filtering through a glass wool plug (2 to 4 cm in length) A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. The aqueous phase or WAF was removed by mid-depth siphoning (the first 75 to 100 mL discarded) to give the 1.0, 3.2, 10, 32 and 100 mg/L loading rate WAFs. Microscopic observations of the WAFs were performed after filtering and showed that no undissolved test item was present.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Strain/Clone: first instar Daphnia magna derived from in-house laboratory cultures.
Adult daphnids were maintained in 150 mL glass beakers containing 100 mL Elendt M7 medium in a temperature controlled room maintaining the water temperature at 18 to 22 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and GEMMA micro 300. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Test temperature:

20 - 21 °C

pH:

7.9 - 9.4 at start; 8.0 - 8.4 at 48 hrs

Dissolved oxygen:

8.9 - 9.0 mg O2/L at start; 8.8 - 9.1 mg O2/L at 48 hrs

Nominal and measured concentrations:

Nominal Loading Rate:
Measured Conc (mg/L): 0 Hr and 48Hr
0 1.0 1.29 1.21
3.2 2.89 2.77
10 9.13 9.13
32 28.1 27.5
100 94.0 92.7

Details on test conditions:

Test System:
Test Vessel: 150 mL glass beakers containing 100 mL Elendt M7 medium
Aeration: no
# organisms/vessel: 5
# vessel per concentration: 4
# of vessels per control: 4

pH Adjustment: No
Photoperiod: 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods.
Light Intensity: between 200 and 1200 lux)

Effect parameters:
Immobilization or adverse reactions to exposure were recorded at 24 and 48 hrs.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

37 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

loading rate WAF

Basis for effect:

mobility

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

47 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

loading rate WAF

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities:
- Observations on body length and weight:
- Other biological observations:
- Mortality of control:
- Other adverse effects control:
- Immobilisation of control:
- Abnormal responses:
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values:
- Effect concentrations exceeding solubility of substance in test medium:

Results with reference substance (positive control):

- Results with reference substance valid? yes
A positive control (Envigo study number LK67NP) used potassium dichromate as the reference item at concentrations of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L.
Exposure conditions for the positive control were similar to those in the definitive test.
Analysis of the immobilization data was carried out using the Trimmed Spearman-Karber method at 24 hours and the Binomial Distribution method at 48 hours. All statistical analysis was carried out using the ToxRat Professional computer software package with results based on the nominal test concentrations and gave the following results:
24 Hours EC50: 0.79 mg/L; 95% confidence limits 0.73 to 0.86 mg/L
48 Hours EC50: 0.75 mg/L; 95% confidence limits 0.56 to 1.0 mg/L

No Observed Effect Concentration 24 Hours: 0.56 mg/L
No Observed Effect Concentration 48 Hours: 0.56 mg/L
Lowest Observed Effect Concentration 24 Hours: 1.0 mg/L
Lowest Observed Effect Concentration 48 Hours: 1.0 mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48 hour EL50 is 37 mg/L

Executive summary:

A study was performed to assess the acute toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

Following preliminary range-finding tests, 20 daphnids (4 replicates of 5 animals) were exposed to WAFs of the test item over a range of nominal loading rates of 1.0, 3.2, 10, 32 and 100 mg/L for 48 hours at a temperature of 20 to 22 °C under static test conditions. The number of immobilized daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours.

Chemical analysis of the fresh test preparations at 0 hours showed measured test concentrations to range from 1.3 to 94 mg/L. Chemical analysis of the aged test preparations at 48 hours showed measured test concentrations to range from 1.2 to 93 mg/L.

The dissolved test item may have been one or several components of the test item. Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

Exposure of Daphnia magna to the test item gave the following results:

Time Point (Hours)	EL50 (mg/L Loading Rate WAF)	95% Confidence Limits (mg/L Loading Rate WAF)	NOEL (mg/L)	LOEL (mg/L)
48	37	Not determined due to the nature of the data	10	32

NOEL=No Observed Effect Loading rate

LOEL=Lowest Observed Effect Loading rate