6-nonyl-1,3,5-triazine-2,4-diamine

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:

2016-02-24 to 2016-03-14

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Analytical samples were taken from all test item concentrations and controls at test start and after 48 hours from aged solutions.
For each sampling also a retain sample was taken.
100 mL samples were taken in 250 mL HDPE bottles without stabilisation.
All samples were stored deep frozen until they were transferred to the analytical laboratory.

Vehicle:

yes

Details on test solutions:

Based on the results of a non GLP range-finding test, the following nominal concentrations were tested in the main test: 4.50, 2.25, 1.13, 0.563 and 0.281 mg/L, solvent control and control.
The necessary amount of test item for preparing the stock solution S1 was weighed on a weighing scoop and transferred to a volumetric flask.
Dimethylformamide was added up to the bench mark and the solution was homogenised by shaking and treated with 10 minutes of ultrasonification.
Lower test solutions were prepared by dilution of the stock solution with DMF.
Final test solution contains 0.1 mL DMF/L

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water Flea
- Strain: Daphnia magna Straus, Clone V
- Source: Eurofins Agroscience Services EcoChem GmbH
- Age at study initiation: max. 24 hours old
- Method of breeding: single culture (1 daphnid per 100 mL) in Elendt M4 medium
- Type and amount of food: single cell green algae (Desmodesmus subspicatus)
- Feeding frequency: at least three times a week
- Feeding during test: none

ACCLIMATION
- Acclimation period: not applicable, freshly hatched daphnids

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

12°dH (214 mg/L CaCO3)

Test temperature:

19.1 ± 0.1 °C

pH:

7.74 ± 0.03

Dissolved oxygen:

9.0 ± 0.1 mg/L

Salinity:

Elendt M4

Nominal and measured concentrations:

The analytical verification of test item concentrations in fish test medium was done by analysing the content of TOC (total organic carbon) in the samples during the test.
Every effort was made to develop an analytical method of the test item. Caprinoguanamine can be determined with isocratic HPLC and detected at
210 nm. But because of the low water solubility and UV-absorption of the test item the determination is not possible in aquatic solutions < 10 mg/L.
Because of missing chromophores Caprinoguanamine shows a too low UV absorption. Therefore, a qualitative detection is not possible at low
concentrations.

Since the stock solution was prepared in DMF the blank value of the solvent control was subtracted from the measured value for the calculation of the content of test item in the test solutions. This resulted partly in negative values, hence no reliable concentration values could be estimated.

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 ml glas vessel, covered with a glass plate
- Material, size, headspace, fill volume: 50 ml
- Aeration: none
- Test procedure: static
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 5
- No. of vessels per vehicle control (replicates): 4


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium


OTHER TEST CONDITIONS
- Photoperiod: 16 hours photoperiod /8 hours darkness daily

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: max. 2.2 requested by the guideline

- Range finding study
- Test concentrations: 4.50, 0.45, 0.045 mg/L
- Results used to determine the conditions for the definitive study:
After 48 hours of exposure no immobilisation was observed in the control, solvent control and up to and including 0.45 mg/L. 100 % of immobilisation was observed at the highest test itemconcentration of 4.50 mg/L.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

2.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % confidence limit 2.47 - 3.87 mg/L

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1.13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

After 24 hours of exposure no immobilisation was observed in the control, solvent control and up to and including 1.13 mg/L. 35 % immobilisation was observed in the test item concentration of 2.25 mg/L and 75 % of immobilisation was observed at the highest test item concentration of 4.50 mg/L.
After 48 hours of exposure no immobilisation was observed in the control, solvent control and up to and including 1.13 mg/L. 60 % immobilisation was observed in the test item concentration of 2.25 mg/L and 100 % of immobilisation was observed at the highest test item concentration of 4.50 mg/L.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: 18 of 20 at 2.0 mg/L after 48 h
- EC50 48h: < 2.0 mg/l

Reported statistics and error estimates:

Probit analysis using linear max. likelihood regression.

Validity criteria fulfilled:

yes

Conclusions:

According to the results of the test, the EC50 (48 h) for immobilisation was determined to be 2.18 mg/L (nominal).
The corresponding NOEC (48 h) was 1.13 mg/L (nominal).

Executive summary:

The 48 hour acute toxicity of Caprinoguanamine to Daphnia magna Straus was studied under static conditions.  Daphnids were exposed to control, solvent control, and test chemical at nominal concentration of 0.281, 0.563, 1.13, 2.25 and 4.50 mg/L for 48 hours.  Mortality/immobilization were observed at 24 and 48 hours. 

The 48 – hour EC₅₀ was 2.18 mg/L (nominal).  The 48 – hour NOEC based on immobilization was 1.13 mg/L (nominal). 

 

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

 

Results Synopsis

Test Organism Age: Daphnia magna Straus > 24 hours

Test Type: Static

EC₅₀:  2.18 mg/L (nominal) 95% C.I.:  2.47 – 3.87 mg/L

Endpoint(s) Effected: immobilisation

Description of key information

The EC50 (48 h) for Caprinoguanamine in Daphnia magna Straus was determined to be 2.18 mg/L test item (nominal).

The corresponding NOEC (immobilization) (48 h) was 1.13 mg/L test item (nominal).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

2.18 mg/L

Additional information

The 48 hour acute toxicity of Caprinoguanamine to Daphnia magna Straus was studied under static conditions.  Daphnids were exposed to control, solvent control, and test chemical at nominal concentration of 0.281, 0.563, 1.13, 2.25 and 4.50 mg/L for 48 hours.  Mortality/immobilization were observed at 24 and 48 hours. 

The 48 – hour EC50 was 2.18 mg/L (nominal).  The 48 – hour NOEC based on immobilization was 1.13 mg/L (nominal). 

 

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