1H-Imidazole, reaction products with epichlorohydrin

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:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Principles of method if other than guideline:

The study was conducted in accordance with the following guidelines:
• OECD Guideline for Testing of Chemicals No. 202, adopted 13. Apr. 2004:
”Daphnia sp., Acute Immobilisation Test“
• Council Regulation (EC) No. 440/2008, Method C.2. “Daphnia sp. Acute Immobilisa-tion Test”, adopted 30. May 2008
• OECD guidance document no. 23, GUIDANCE DOCUMENT ON AQUATIC TOXIC-ITY TESTING OF DIFFICULT SUBSTANCES AND MIXTURES, adopted 14. Dec. 2000

GLP compliance:

yes (incl. QA statement)

Remarks:

GLP-Conformity All procedures will be according to the principles of GLP (Chemikaliengesetz §19a and §19b and annexes 1 and 2 from 28. Aug. 2013, published in Federal Law Gazette, Germany (BGBl) No. 55/2013 as of 06. Sep. 2013, and further revisions).

Test material

Test material information

Constituent 1

additive 1

Test material form:

liquid

Details on test material:

- State of aggregation: not applicable
- Activation: not required
water as a additive

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: The test material is representative of the registered substance
- Expiration date of the lot/batch: not relevant
- Purity test date: not relevant

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature in the dark
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: soluble

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none
- Preliminary purification step (if any): none

Sampling and analysis

Analytical monitoring:

yes

Remarks:

TC

Details on sampling:

The content of the test item in the test solutions was estimated by calculation based on the organic carbon content of the test item and DOC measurement. DOC (dissolved organic carbon) was determined as TC (total carbon) minus IC (inorganic carbon).
The carbon content of the test item was determined in a non-GLP pre-test by DOC measurement of a 1 g/L stock solution of the test item.

Analyses of the dissolved organic carbon were made by TC and IC measurement using the carbon analyser TOC multi N/C 2100S, Analytik Jena. Each sample was measured in duplicate or triplicate, respectively (depending on the variation between the measured values). The carbon analyser was calibrated with freshly prepared reference solutions containing potassium hydrogen phthalate (TC), sodium hydrogen carbonate and sodium carbonate (IC) once a month. After every start, quality control samples were measured.

Reference Items for Carbon Determination
Potassium hydrogenphthalate for TC (Batch no. MKBS1485V, p.A., content 99.99 %), Na2CO3 (Batch no. 247259189 , p.A., content 99.8 %) and NaHCO3 (Batch no. 217156106 , p.A., content 99.9 %) for IC.

Test solutions

Vehicle:

no

Details on test solutions:

Positive Control
Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9) was used as positive control in a current reference study (201701R201). The 24h-EC50i value was determined as 1.6 mg/L. The value was within the requested range for the 24h-EC50 between 0.6 - 2.1 mg/L of po-tassium dichromate K2Cr2O7 as specified in the OECD guideline. Since the fits showed sufficient statistical correspondence of the data with the dose-response-equation, this reference study can be considered valid
Dilution water (Daphnia medium) with the following specification was used (stated in SOP 11800201)
Dilution water specification
Parameter Concentration in mg/L
CaCl2*2H2O 293.80
MgSO4*7H2O 123.30
NaHCO3 64.80
KCl 5.80
Resulting hardness in mmol/L: 2.502
Resulting hardness in mg CaCO3/L: 250
Deviations from the nominal weighted loads were less than 5%. Exact values were recorded in the raw data. After preparation, the dilution water was aerated and the pH was measured. The pH was 7.8.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Specification
Species/Strain                              Daphnia magna Straus / Origin Umweltbundesamt Berlin, In-house breeding since 27. September 2007
Sex                                                 female
Age                                                 between 0 and 24 hours
Animal Husbandry
Daphnia magnais bred in the LAUS GmbH throughout the year. The animals are kept for the use in toxicity tests. They multiply by parthenogenesis, thus being genetically identical.
Vessels                                          preserving glasses, nominal volume 2 L
Medium                                         M4-Medium (recipe of Elendt)
Food                                               green algae (Desmodesmus subspicatus)
Medium renewal                          twice a week
Photo period                                 16/8 hours, using neon tubes
Temperature                                 20±2 °C
Selection of Animals
20 ± 4 hours before the start of the test, the adult animals are separated from the young. 1 ± 0.5 hours before the start of the test, the adults are caught with the help of a glass tube, and the newborn daphnia, aged between 0 and 23 hours, are sieved from the medium and immediately placed into a glass beaker containing dilution water.
Switching from M4-medium (husbandry) to dilution water (test) has been shown not to cause any detrimental effects for test daphnia.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

Duration 48 ± 1 hours

Post exposure observation period:

Observation times 24 ± 1 and 48 ± 1 hours

Test conditions

Hardness:

Resulting hardness in mmol/L: 2.502
Resulting hardness in mg CaCO3/L: 250

Test temperature:

Temperature 20.4 - 21.8 °C. range 18.0 – 22.0 °C, within one test, temperature may not vary more than 2 °C e.g. 19.0 – 21.0 °C

pH:

The dilution water will be prepared using demineralised water. After preparation, the dilution water is aerated, the pH will be measured and if necessary adjusted to 7.8 ± 0.2. in Experiment the pH was 7.8 - 8.0

Dissolved oxygen:

Concentration of dissolved oxygen at the end of the test must be at least 3 mg/L.
O2-Concentration in mg/L:
At start: 9.1, at end: 8.9- 9.0

Nominal and measured concentrations:

measured as DOC

Details on test conditions:

The study is conducted in accordance with the following guidelines:
• OECD Guideline for Testing of Chemicals No. 202, adopted 13. April 2004: ”Daphnia sp., Acute Immobilisation Test“
• Council Regulation (EC) No. 440/2008, Method C.2. “Daphnia sp. Acute Immobilisa-tion Test”, adopted 30. May 2008
• OECD guidance document no. 23, GUIDANCE DOCUMENT ON AQUATIC TOXIC-ITY TESTING OF DIFFICULT SUBSTANCES AND

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7, CAS No. 7778-50-9)

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.046 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.089 - <= 0.148 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

>= 0.197 - <= 0.306 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

• Immobilisation in the controls may not exceed 10 %.
Immobilisation in the controls was 0 %.
• The concentration of dissolved oxygen at the end of the test must be at least 3 mg/L. The lowest concentration of dissolved oxygen at the end of the test was 8.9 mg/L.

Results with reference substance (positive control):

Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9) was used as positive control in a current reference study (201701R201). The 24h-EC50i value was determined as 1.6 mg/L. The value was within the requested range for the 24h-EC50 between 0.6 - 2.1 mg/L of po-tassium dichromate K2Cr2O7 as specified in the OECD guideline. Since the fits showed sufficient statistical correspondence of the data with the dose-response-equation, this reference study can be considered valid.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Biological Results Test Item
Parameter Value 95%-confidence interval
24h EC50 0.246 mg/L 0.197 - 0.306 mg/L
48h EC50 0.116 mg/L 0.089 - 0.148 mg/L
48h NOEC 0.046 mg/L --
48h LOEC 0.1 mg/L --

Executive summary:

The study was performed according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) using a concentration  0.046 to 1 mg/L (nominal). with 20 Daphnia magna for test concentration and blank controls in a static test system. After 24 and 48 hours, the immobilised Daphnia were counted.

All concentrations showed toxicity between 10 and 100% immobilisation. None of the animals was immobilised in the blank control.

NOEC = 0.046 mg/L

LOEC = 0.1 mg/L

EC50 = 0.116 mg/L

The results of the test are considered valid.