Carbazamidine sulphate

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Test guideline 3

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 1: Static Method)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Guanidinium phosphate (1:1)

EC Number:

226-551-9

EC Name:

Guanidinium phosphate (1:1)

Cas Number:

5423-22-3

Molecular formula:

CH5N3.H3O4P

IUPAC Name:

Guanidium phosphate (1:1)

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Blank control and 100 mg/L
- Sampling
Frequency: at t=0 h and t=96 h
Volume: 1 mL
- Sample storage conditions before analysis: Samples were transported on dry ice to TNO Triskelion (P.O. Box 844, 3700 AV, Zeist, The Netherlands) and were stored in a freezer until analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Preparation of test solutions started with a nominal concentration of 100 mg/L applying a short period of vigorous shaking to accelerate the dissolving of the test substance in the test medium. The lower test concentrations for the range-finding test were prepared by subsequent dilutions of the 100 mg/L concentration in test medium.
- Controls: Blank control containing test medium without test substance or other additives
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The final test solutions were all clear and colourless.

Test organisms

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, the Netherlands
- Length at study initiation (mean and SD): 2.3±0.3 cm (final test)
- Weight at study initiation (mean and SD): 0.29±0.10 g (final test)
- Feeding during test: None

ACCLIMATION
- Acclimation period: At least 12 days after delivery
- Acclimation conditions (same as test or not): Yes
- Feeding frequency and type of food: Daily with pelleted fish food
- Health during acclimation (any mortality observed): In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

180 mg CaCO3 per litre

Test temperature:

Final test: 21.4 - 22.5°C

pH:

Final test
Blank control: 7.5-7.7
100 mg/L: 6.7-7.3

Dissolved oxygen:

Final test (O2 (mg/L)
Blank control: 6.6 - 9.2
100 mg/L: 7.2 - 9.2

Nominal and measured concentrations:

Nominal (final test): 0 (blank control); 100 mg/L.
Measured (final test)
t=0h: <0.08 mg/L* (blank control) and 85 mg/L (100 mg/L)
t=96h: <0.08 mg/L* (blank control) and 97 mg/L (100 mg/L)
*) LOD was 0.08 mg/L (lowest calibration point).

Details on test conditions:

TEST SYSTEM
- Test vessel: 10 litres, all-glass, containing 9 litres of test solution
- Aeration: None
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.23 g fish per litre

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap-water purified by reverse osmosis
- Culture medium different from test medium: No (adjusted ISO medium). However, the pH of culture medium was 7.9. Therefore fish to be exposed to the 100 mg/L concentration were acclimatized to a pH of 7.0 before the start of the test.
- Intervals of water quality measurement (dissolved oxygen content, pH and temperature): Daily in all vessels, beginning at the start of the test (day 0).

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light: 8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): At 3h:15min, 24, 48, 72 and 100 h following the start of exposure. In addition, every afternoon from day 0 and every morning from day 1 to observe for any dead or severely distressed fish.

TEST CONCENTRATIONS
- Spacing factor for test concentrations (final test): Limit test (100 mg/L)
- Range finding study
- Test concentrations: blank control, 0.1, 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 33 mg/L

Nominal / measured:

nominal

Conc. based on:

other: guanidine content (32.7% w/w)

Basis for effect:

mortality (fish)

Details on results:

- Measured concentrations: The measured concentration was in agreement with nominal and remained stable during the test period (85-97%). Given this result the LC50-value was based on the nominal test concentration.

- Biological results: No mortality or clinical effects were observed during the test period.

- Acceptability of the test:
1. No mortality was observed in the control at the end of the test.
2. Test conditions were maintained constant throughout the test.
3. The dissolved oxygen concentration has been at least 60% of the air saturation value throughout the test (>5 mg/L at 22°C).
4. The concentrations of the substance being tested have been at least 80% of the nominal concentration throughout the test.

Results with reference substance (positive control):

- Pentachlorophenol: The 96h-LC50 for carp was 0.17 mg/L (95% confidence interval: 0.15 and 0.27 mg/L).

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 96h-LC50 for carp exposed to Guanidine phosphate (1:1) was >100 mg/L.

Executive summary:

In a 96-h acute toxicity study conducted according to OECD guideline 203, carp (Cyprinus carpio) were exposed to Guanidine phosphate (1:1) under static conditions at the following nominal concentrations (limit test): 0 (blank control) and 100 mg/L. Test substance induced no visible or lethal effects in carp at a nominal exposure concentration of 100 mg/L. The 96h-LC50 was >100 mg/L (corresponding to >33 mg guanidine/L) based on analytically confirmed nominal concentrations. The study is considered to be reliable without restrictions.