Reaction product of: 2,4-diamino-6-[2-(2-methyl-1H-imidazol-1-yl)ethyl]-1,3,5-triazine and cyanuric acid

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:

07 - 19 Mar 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesanstalt für Umwelt, Messungen und Naturschutz, Karlsruhe, Germany (10 Dec 2015)

Test material

Test material information

Constituent 1

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control and highest test item concentration at test start, after 24 h (fresh and aged solutions) and after 48 h (aged solutions).
- Sampling method: Two samples were taken and a retain sample was also taken for each sampling.
- Sample storage conditions before analysis: Deep frozen (≤ 18 °C)

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A stock solution of 100 mg/L test item was prepared by adding the necessary amount of test item to the test medium and treating the solution with 10 min ultrasonication. Lower test solutions were prepared by dilution of the stock solution or appropriate solution with test medium. The preparation procedure was repeated after 24 h.
- Differential loading: No
- Controls: Same test medium and exposure conditions but without test item.
- Evidence of undissolved material: The stock solution was clear and transparent.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Strain/clone: Clone V
- Source: Originally purchased from the Federal Environment Agency (Berlin, Germany) and bred in the in-house laboratory.
- Breeding conditions: D. magna was bred as single culture (1 daphnid per 100 mL) in Elendt M4 medium. The pH-value of the aerated water was within a range of 6.0 - 9.0. The dissolved oxygen was above 60% saturation and the total hardness 140 - 250 mg/L (CaCO3), corresponding to 7.8 - 14 °dH. The animals were fed with single cell green alga (D. subspicatus) at least 3 times a week. The daphnids were reared at a temperature of 20 ± 2 °C in a climatic chamber with 16 h illumination and 8 h darkness. The medium was changed 3 times per week. A pipette was used to separate the young daphnids from the adults.
- Age of daphnids at test start: freshly hatched, < 24 h old
- Feeding during test: No feeding during test

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

140 - 250 mg/L = 7.8 - 14 °dH (Elendt M4 medium)

Test temperature:

18.8 - 18.9 °C (mean)

pH:

7.73 - 7.90 (mean)

Dissolved oxygen:

9.1 - 9.2 mg/L (mean)

Nominal and measured concentrations:

Control, and 100 mg/L (nominal)
< LOQ, and 102 mg/L (mean measured)

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass vessels filled with ~ 50 mL test solution and covered with a glass plate to reduce evaporation.
- Renewal rate of test solution: 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 1 daphnid per ~10 mL test solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium
- Culture medium different from test medium: Culture medium same as test medium
- Intervals of water quality measurement: The test temperature and the pH values as well as the oxygen concentration of the test solutions were measured at all concentration levels at 0 h (fresh medium), 24 h (fresh and aged medium) and at 48 h (aged medium) in one separate replicate per test item concentration without test organisms.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h dark

EFFECT PARAMETERS MEASURED:
- Immobilization: after 0, 24, and 48 h

RANGE-FINDING STUDY
- Test concentrations: Control, 0.01, 0.1, 1.0, 10.0, and 100 mg/L
- Results used to determine the conditions for the definitive study: Results are not reported.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: No
- Mortality of control: 0%
- Any observations that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes, the test was performed in accordance with OECD guideline and the results fell within the historical data generated with the reference item potassium dichromate at the testing facility.
- Relevant effect levels: EC50 (24 h) was between 1.00 and 2.00 mg/L
- Other: 100% immobilization after 48 h (2.0 mg/L), 65% immobilization after 48 h (1.0 mg/L)

Reported statistics and error estimates:

The NOEC was established based on the highest concentration at which the immobilisation is not higher than the allowed control immobilisation (≤ 10%). Due to a lack of immobilisation, the EC50 could not be statistically determined.

Any other information on results incl. tables

VALIDITY CRITERIA

The study met the validity criteria laid down by the guideline (Table 1).

Table 1: Validity criteria for OECD 202.

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	The control immobilization was 0%.	Yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	The dissolved oxygen concentration in this test was ≥ 9.0 mg/L at the end of the test.	Yes

 

ANALYTICAL RESULTS

The measured test item concentration was between 92 and 101% of nominal in fresh samples with a mean initial concentration of 97% of nominal. In aged samples, the measured test item content ranged from 101 – 103% of nominal with a mean measured concentration of 102% of nominal (Table 2). Therefore, the toxicological assessment was based on the nominal concentrations.

Table 2. Measured test item concentrations.

nominal concentration	sampling	test item
[mg/L]	[h]	[mg/L]	% of nominal
Control	0  (fresh medium)	n.d.	-
	24 (aged medium)	n.d.	-
	24 (fresh medium)	n.d.	-
	48 (aged medium)	n.d.	-
100	0  (fresh medium)	101	101
	24 (aged medium)	101	101
	24 (fresh medium)	92.4	92
	48 (aged medium)	103	103

- = not calculated; n.d. = not detectable; LOQ = 1.00 mg/L

 

BIOLOGICAL RESULTS

After 48 h no immobilization was observed up to and including 10.0 mg/L. No immobilization higher than the allowed control immobilization was observed at 100 mg/L (Table 3).

 

Table 3. Immobilization after 48 h

	nominal test item concentration [mg/L]
	control	0.01	0.10	1.00	10.0	100
rep 1	0	0	0	0	0	1
rep 2	0	0	0	0	0	0
rep 3	0	0	0	0	0	1
rep 4	0	0	0	0	0	0
Σ	0	0	0	0	0	2
%	0	0	0	0	0	10

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.