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

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-02-22 to 2016-03-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Analytical samples were taken in 24 h intervals from fresh and aged test solutions.
For each sampling also a retain sample was taken.
100 mL samples were taken in 250 ml HDPE bottles without stabiliser and stored deep frozen until they were transferred to the analytical laboratory.

Analytical samples taken at t = 0 h and t = 96 h were analysed from control, solvent control, 0.107, 0.235 and 0.517 mg/L to determine the total organic carbon.
Additionally, samples taken at t = 0 h and t = 24 h were analysed from 1.14 and 2.50 mg/L.

Vehicle:

yes

Details on test solutions:

Based on the results of the range-finding test, the following nominal concentrations were tested in the main test: 0.107, 0.235, 0.517, 1.14 and
2.50 mg/L, solvent control and control.
The necessary amount of test item for preparing the stock solution was weighed in a 20 mL glas vial. 5 mL of Dimethylformamide (DMF) was added
and care was taken that the stock solution was well homogenized by shaking and ultrasonification for 20 minutes.
The lower test solutions were prepared by dilution of the stock solution with DMF.
A defined volume of the prepared solutions were transferred into each test vessels containing 15 L of test medium and stirred.
Application volume of DMF per test vessel 1.5 mL.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow Trout
- Strain: Salmonidae
- Source: Forellenzucht Peter Störk, D-88348 Bad Saulgau, Germany
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): 4 - 6 cm
- Weight at study initiation (mean and range, SD):
- Method of breeding:
- Feeding during test
none

ACCLIMATION
- Acclimation period: > 12 days
- Acclimation conditions (same as test or not): yes
- Type and amount of food: granular rearing food with approx. 2 % of their body-weight
- Feeding frequency: each day
- Health during acclimation (any mortality observed): No mortality above 5 %

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

12°dH corresponding to 214 mg CaCO3/L

Test temperature:

15.6 - 16.0 °C

pH:

7.78 - 8.50

Dissolved oxygen:

≥ 85 % of air saturation

Conductivity:

529 μS/cm

Nominal and measured concentrations:

0.107, 0.235, 0.517, 1.14 and 2.50 mg/l, (nominal) solvent control and control.

Details on test conditions:

TEST SYSTEM
- Test vessel: 25 L glass aquaria filled with 15 L of test solution
- Aeration:
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: max. loading of 1 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated drinking water and deionised water
- Total organic carbon:
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:
- Ca/mg ratio:
- Conductivity: 529 μS/cm.
-
- Intervals of water quality measurement:

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 16 hours daily
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Fish were observed at 0, 4, 24, 48, 72 and 96 hours after test start.
Visible abnormalities as: loss of equilibrium, swimming behaviour, respiratory function, pigmentation and all other observed events were recorded.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Justification for using less concentrations than requested by guideline: Number of concentrations and spacing factor according to guideline.
-
Range finding study
- Test concentrations: 0.045, 0.45, 4.5 mg/L
- Results used to determine the conditions for the definitive study: 0.045 mg/l no mortality; 0.45 mg/L 33 % and 4.5 mg/L 100 % mortality

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.768 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.517 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.235 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: sublethal effects

Details on results:

No sublethal effects were observed in the control, solvent control and at the test item concentrations up to and including 0.235 mg/L.

At 0.517 mg/L one fish had difficulties with maintenance of equilibrium and one fish was upside down with loss of equilibrium, showing only movement of gills as a sign of life after 24 h. After 48 h three fish had difficulties with maintenance of equilibrium. After 72 h two fish had difficulties with maintenance of equilibrium and one fish was upside down with loss of equilibrium, showing only movement of gills as a sign of life.
After 96 h one fish was upside down with loss of equilibrium, showing only movement of gills as a sign of life.

At 1.14 mg/L three fish were upside down with loss of equilibrium, showing only movement of gills as a sign of life after 4 h. Four fish were found dead. After 24 h all fish were found dead.

At the highest test item concentration of 2.50 mg/L all fish were found dead after 4 h.

The average weight of the test organisms was 1.22 ± 0.30 g; the average length was 53 ± 4 mm.

Reported statistics and error estimates:

The LC50-value after 4 h was calculated using Trimmed Spearman-Kaerber procedure (ANELLIS & WERKOWSKY, 1968). Only concentrations within a clear dose response were used for calculations. For data evaluation the statistical programme ToxRat Professional 3.2.1 was used.
The level of significance for all statistical procedures was set to α = 0.05.
The LC50-values after 24, 48, 72 and 96 h were calculated as geometric mean between the highest concentration without any mortality and the lowest concentration producing 100 % mortality.

The NOEC (mortality) was established based on the highest test concentration at which no mortality above the allowed control mortality was observed.

Validity criteria fulfilled:

yes

Conclusions:

According to the results of the test, the LC50 (96 h) of the test item was determined to be 0.768 mg/L test item (nominal).
The corresponding NOEC (mortality) (96 h) was 0.517 mg/L test item (nominal).

Executive summary:

In a 96-h acute toxicity study according to OECD guideline 203, Rainbow Trout Oncorhynchus Mykiss were exposed to Caprinoguanamine at nominal concentrations of 0, solvent control (DMF), 0.107, 0.235, 0.517, 1.14 and 2.50 mg/L under static conditions. 

The 96-h LC50 was 0.768 mg /L (nominal).  The NOEC values, based on mortality/sublethal effects, were 0.517 and 0.235 mg/L (nominal), respectively. 

No sublethal effects were observed in the control, solvent control and at the test item concentrations up to and including 0.235 mg/L. At 0.517 mg/L one fish had difficulties with maintenance of equilibrium and one fish was upside down with loss of equilibrium, showing only movement of gills as a sign of life after 24 h. After 48 h three fish had difficulties with maintenance of equilibrium. After 72 h two fish had difficulties with maintenance of equilibrium and one fish was upside down with loss of equilibrium, showing only movement of gills as a sign of life. After 96 h one fish was upside down with loss of equilibrium, showing only movement of gills as a sign of life.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for fish acute toxicity study.

 

Results Synopsis

 

LC50:  0.768 mg /L (nominal)

NOEC (mortality): 0.517 mg/L (nominal)

NOEC (sublethal effects): 0.235 mg/L (nominal)

 

 

Description of key information

The LC50 (96 h) for Caprinoguanamine in Rainbow Trout Oncorhynchus Mykiss was determined to be 0.768 mg/L test item (nominal).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

0.768 mg/L

Additional information

In a 96-h acute toxicity study according to OECD guideline 203, Rainbow Trout Oncorhynchus Mykiss were exposed to Caprinoguanamine at nominal concentrations of 0, solvent control (DMF), 0.107, 0.235, 0.517, 1.14 and 2.50 mg/L under static conditions. 

The 96-h LC50 was 0.768 mg /L (nominal).  The NOEC values, based on mortality/sublethal effects, were 0.517 and 0.235 mg/L (nominal), respectively. 

No sublethal effects were observed in the control, solvent control and at the test item concentrations up to and including 0.235 mg/L. At 0.517 mg/L one fish had difficulties with maintenance of equilibrium and one fish was upside down with loss of equilibrium, showing only movement of gills as a sign of life after 24 h. After 48 h three fish had difficulties with maintenance of equilibrium. After 72 h two fish had difficulties with maintenance of equilibrium and one fish was upside down with loss of equilibrium, showing only movement of gills as a sign of life. After 96 h one fish was upside down with loss of equilibrium, showing only movement of gills as a sign of life.