1-benzyl-3-carbamoylpyridinium chloride

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 - 29 September 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

13 April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

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

OTHER SPECIFICS: White Powder

Analytical monitoring:

yes

Details on sampling:

- Sampling method: Samples of 1 mL were taken from two replicates of treatment A, C, E and NC at 0 h, 24 h and 48 h, and filled separately into 1,5 mL Eppendorf vials.
- Sample storage conditions before analysis: The samples were stored in the freezer at ≤ -18 °C. Of each treatment and each sampling time one of the samples was sent to the analytical laboratory at the test site of biochemA GmbH. All other samples were stored as retain samples in the freezer (<-18°C) until finalization of the study.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The stock solution is prepared by adding 146.4 mg (0 h), respectively 146.5 mg (24 h) of the test item to 500 mL M4 medium by stirring with a magnetic stirrer for about 5 minutes. The further test concentrations are prepared by diluting the stock solution with M4 medium
- Eluate: M4-Medium
- Controls: M4-Medium

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna Straus
- Source: clone breeding of the Federal Environment Agency, department IV 2.4, Berlin, and has been cultivated at Hydrotox GmbH since October 2012
- Age at study initiation: 0.25 – 22 h
- Feeding during test: none


BREEDING
- Food type: Desmodesmus subspicatus; fed on workdays
- Amount: Daily 0.1 mg C per daphnid
- Light/dark cycle: 16/8 h
- Temperature: 20 +/- 2 °C
- Housing: 10 animals/ 200 mL
- Medium: M4 (ELENDT), transfer twice per week into fresh medium

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

19.3 – 20.6 °C

pH:

7.7 – 7.9 in the control and 7.5 – 7.9 in the test item treatments

Dissolved oxygen:

8.0 – 8.2 mg/L in the control and 8.0 – 8.3 mg/L in
the test item treatment

Nominal and measured concentrations:

0 (NC), 12.5 (A), 27.5 (B), 62.5 (C), 133.1 (D), 292.8 (E) mg/L.
The concentrations of 1-Benzyl-3-carbamoyl-pyridinium, chloride in the control were below the limit of quantification. The measured test item concentrations were within the range of ± 10% of the nominal concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers, 50 ml, Schott, Mainz
- Fill volume: 20 mL
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): once after 24h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

OTHER TEST CONDITIONS
- Photoperiod:16/8 h

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Mobility after 24 and 48 h

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: A preliminary test without GLP was performed before start of this GLP-study. Nominal concentrations of 10 and 100 mg/L test item were tested and showed 0 and 45 % immobility after 48 h exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Test concentrations: 0 (NC), 12.5 (A), 27.5 (B), 62.5 (C), 133.1 (D), 292.8 (E) mg/L
- Range finding study. yes

Reference substance (positive control):

no

Remarks:

Quality assurance takes place regularly by testing the sensitivity of the test organisms to potassium dichromate. The recent quality testing was performed in June 2016 with EC50 (24h) = 1.30 mg/L (CL 95 %: 1.20 - 1.42 mg/L) and EC50 (48h) 0.82 mg/L.

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

60.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

other: EC20

Effect conc.:

113.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

377 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

LOEC

Effect conc.:

292.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

133.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

42 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

other: EC20

Effect conc.:

69.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

181.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

292.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

133.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The EC10 and EC20 values for both evaluation dates (after 24 h and 48 h) are below the respective LOEC values for the same data. This can be explained by the differing statistical evaluation. The EC values are derived from a calculated concentration-effect curve. The LOECs are calculated as threshold concentration and have to differ significantly from the control group. Hence, though 15 % individuals are reported to be immobile at 133 mg/L after 48 h, this amount does not vary significantly from the control and is therefore classified by the statistical evaluation as NOEC.
The immobility in the control is reported as 0 % after 24 h and as 5 % after 48 h.

Results with reference substance (positive control):

n.a.

Reported statistics and error estimates:

The effect occurrence after 24 h and 48 h exposure was calculated. ECx and LOEC/NOEC were determined with the statistical software ToxRat Professional 3.2.1 (ToxRat Solutions GmbH, Alsdorf, Germany), as far as mathematically possible.

The reported immobility during the test is presented in Table 4. The values for the calculated ECx, LOEC and NOEC are displayed in Table 5. The results are also visualized in figure 1 and 2 in the illustration part of the present documentation part.

Table 4: Immobility [%] after 24 h and 48 h exposure

Nominal test item concentration [mg/L]	Immobility [%]
	24 h	48 h
NC	0	5
12.5	5	10
27.5	0	0
62.5	15	10
133.1	0	15
292.8	60	90

Table 5: Effect concentrations

(Lowest/No Observed) Effect concentrations	Nominal test item concentration [mg/L]
	24 h	48 h
EC10	60.5	42.0
EC 20	113.4	69.4
EC 50	377.0	181.7
LOEC	292.8	292.8
NOEC	133.1	133.1

The 95 % confidence limits could not be calculated by the statistical evaluation with ToxRat Professional 3.2.1.

Validity criteria fulfilled:

yes

Conclusions:

The 48-h EC50 of 1-Benzyl-3-carbamoyl-pyridnium, chloride to Daphnia magna is 181.7 mg/L.

Executive summary:

The 48-h-acute toxicity of 1-Benzyl-3-carbamoyl-pyridinium, chloride to Daphnia magna was studied under semi-static conditions in accordance with OECD Guideline 202. Daphnids were exposed to the test chemical at nominal concentrations of 0, 12.5, 27.5, 62.5, 133.1 and 292.8 mg/L for 48 h. Immobilisation was observed daily. The 48-h EC₁₀ was 42.0 mg/L. The 48-h EC₅₀ was 181.7 mg/L. The 48 h NOEC based on immobilisation exceeds or equals 133.1 mg/L and the LOEC equals 292.8 mg/L. The values of the EC₁₀ and NOEC rsp. LOEC vary, due to different statistical evaluation methods. As the 48-h EC₅₀ exceeds the threshold of 1 mg/L, the substance is not classified to be toxic according to the Regulation (EC) No 1272/2008, Annex 1, table 4.1.0.

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

Results Synopsis

Test organism age: Neonates

Test type: Semi-static

EC₅₀: 181.7 mg/L (95 % C.I.: n.d.)

NOEC ≥ 133.1 mg/L

Endpoint(s) effected: Immobilisation

Description of key information

The 48 h acute toxicity of 1-Benzyl-3-carbamoyl-pyridinium, chloride to aquatic invertebrates (Daphnia magna) was investigated in a study, according to GLP and OECD 202 under semi-static conditions. The measured test item concentrations were within the range of ± 10% of the nominal concentrations. The Effect concentrations are as follows: 24 h EC₁₀ (nominal): 60.5 mg/L (95%-CI: n.d.); 24 h EC₅₀ (nominal): 377.0 mg/L (95%-CI: n.d.) 24 h LOEC = 292.8 mg/L; 24 h NOEC ≥ 133.1 mg/L; 48 h EC₁₀ (nominal): 42.0 mg/L (95%-CI: n.d.); 48 h EC₅₀ (nominal): 181.7 mg/L (95%-CI: n.d.); 48 h LOEC: 292.8 mg/L; 48 h NOEC ≥ 133.1 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

181.7 mg/L