Reaction mass of 2,6-dimethylpyridinium chloride and 3-methylpyridinium chloride and 4-methylpyridinium chloride

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 2,6-dimethylpyridinium chloride and 3-methylpyridinium chloride and 4-methylpyridinium chloride

EC Number:

941-122-2

Molecular formula:

not yet assigned

IUPAC Name:

Reaction mass of 2,6-dimethylpyridinium chloride and 3-methylpyridinium chloride and 4-methylpyridinium chloride

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: LC-MS/MS

Test solutions

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The stock solution (100 mg/L of the test item were weighed out) was freshly prepared in a measuring flask with dilution water before the start of the exposure (at 0 h).
- Controls: Dilution water without test item tested under the same conditions as the test groups. 4 replicates, each with 5 daphnids.
- Evidence of undissolved material: The stock solution was prepared in a measuring flask and mixed thoroughly by inverting the measuring flask several times until the test item was completely dissolved.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (Straus)
- Method of breeding: In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20  2°C, in an incubator, 16 h illumination, light intensity of max. 20 µExm-2 x s-1
- Feeding during test
- Food type: mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus
- Amount: algae cell density of > 10^6 cells/mL
- Frequency: 5 times per week ad libitum

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

NaHCO3 64.8 mg/L

Test temperature:

18 - 19 °C

pH:

test start: 6.2-7.61
test end: 6.48-6.98

Dissolved oxygen:

test start: 8.8-9.08 mg/L
test end: 7.82-8.62 mg/L

Nominal and measured concentrations:

nominal: 100 mg/L
The measured concentration of the test item at the start of the exposure (0 h) was 100 % of the nominal value. At the end of the exposure (48 h), the measured concentration of the test item was 106 % of the nominal value.

Details on test conditions:

TEST SYSTEM
- Test vessel:
-Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity, loosely covered with laboratory film (PARAFILM®)
- Test volume: 20 mL in each test replicate
- Aeration: no
- No. of organisms per vessel: 5 daphnids
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: double distilled water

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 20 µE x m^-2 x s^-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): immobilisation was monitored one and two days after exposure

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:

- Mortality of control: all daphnids were mobile in the control during the 2 days of exposure.
- Any observations that might cause a difference between measured and nominal values: the test substance was soluble in medium, the test concentrations were present as a clear solution.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50/LC50: EC50 (24 h): 2.01 mg/L (CI 1.71 - 2.26 mg/L) with 95 % confidence limits (CI)

Any other information on results incl. tables

Table3:      Immobilisation Rates after 24 and 48 h ofExposure in the Definitive Test (n = 20, divided into 4 replicates with 5 daphnids each)

Pyridinderivate 1_HCl Nominal test item concentration [mg/L]	IMMOBILISATION [%]
	24 h					48 h
	Replicates					Replicates
	1	2	3	4	MV	1	2	3	4	MV
100	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

MV   = mean value

The validation criteria were fulfilled as there were no immobilised daphnids in the control group and oxygen saturation, temperature, pH value as well as measured values for the test medium were in the required ranges.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

At the nominal concentration of 100 mg/L of the test item Pyridinderivate 1_HCl no effects were observed on Daphnia magna.