Calcium 3-hydroxy-4-[(1-sulphonato-2-naphthyl)azo]-2-naphthoate

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:

03 Nov 2017 - 02 Feb 2018

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Test item analysed but

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control and Saturated Solution prepared at 100 mg/L
- Sampling method: 3.0 mL at t=0 h, t=24 h, and t=96 h
- Sample storage conditions before analysis: Samples were stored in a freezer (≤-15°C) until analysis at the analytical laboratory of the Test Facility.
- The filter containing the undissolved residue after test solution preparation was kept for possible analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test item was not completely soluble in test medium at the loading rate initially prepared. Preparation of test solutions started with a loading rate of 100 mg/L applying a two-day period of magnetic stirring to reach the maximum dissolution of the test item in medium. The obtained aqueous mixture was filtered through a 0.45 µm membrane filter (RC55, Whatman) and the Saturated Solution (SS) was used as the highest test concentration.
- Controls: Test medium without test item or other additives.
- Evidence of undissolved material: no, all test solutions were clear and colorless at the end of the preparation procedure.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Strain: Cyprinus carpio (Teleostei, Cyprinidae) Linnaeus, 1758
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands.
- Length at study initiation: 3.1 ± 0.4 cm (final test); 2.7 ± 0.3 cm (range-finding test)
- Weight at study initiation: 0.51 ± 0.21 g (final test); 0.24 ± 0.08 g (range-finding test)
- Characteristics: F1 from a single parent-pair bred in UV-treated water.

ACCLIMATION
- Acclimation period: At least 12 days after delivery

FEEDING DURING TEST
- No feeding from 24 hours prior to the test and during the total test period

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

180 mg CaCO3 per litre

Test temperature:

21-22 °C

pH:

7.6-8.1

Dissolved oxygen:

5.6-9.9 mg/L

Nominal and measured concentrations:

Nominal loading rate: saturated solution at 100 mg/L
Measured concentration: <0.010 mg/L throughout the test, see Table 3 in "Any other information on results incl. tables"

Details on test conditions:

TEST SYSTEM
- Test vessel: 9-L, all-glass, containing 8 L of test solution
- Aeration: no
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.45 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium with a hardness of 180 mg CaCO3 per litre and a pH of 7.7 ± 0.3, formulated using RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands) with the following composition:
CaCl2.2H2O: 211.5 mg/L
MgSO4.7H2O: 88.8 mg/L
NaHCO3: 46.7 mg/L
KCl: 4.2 mg/L
- Intervals of water quality measurement: Dissolved oxygen, pH and temperature: daily in all vessels with surviving fish, beginning at the start of the test (day 0). In addition, pH and temperature were measured before transferring the fish to the test system.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED:
Mortality and other effects: At 2½, 24, 48, 72 and 96 hours following the start of exposure. In addition, every afternoon from day 0 to observe for any dead or severely distressed fish.

TEST CONCENTRATIONS
- Range finding study:
* Test concentrations: 1.0, 10 and 100% of a SS prepared at 100 mg/L (3 fish/concentration)
- Results used to determine the conditions for the definitive study: No mortality or clinical effects were observed at any of the test concentrations during the test period.

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol (Feb 2017)

Key result

Duration:

96 h

Dose descriptor:

LC50

Basis for effect:

mortality (fish)

Remarks on result:

other: LC50 is considered to be above the maximum solubility of the test item in test medium

Details on results:

No mortality or clinical effects were observed at both the SS and the control during the test period (see Table 2 in "Any other information on results incl. tables". The responses recorded in the SS were in agreement with the results of the range-finding test.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- The 96h-LC50 for carp exposed to PCP was 0.32 mg/L (95 % confidence interval between 0.22 and 0.46 mg/L). This is within the range of sensitivities generally observed during the past 30 years (0.10 - 0.46 mg/L)

Sublethal observations / clinical signs:

Table 3          
Final Test: Test Samples

Time of sampling [hours]	Date of sampling	Date of analysis^1	Percentage of SS^2 [%]	Analyzed concentration [mg/L]	Relative to initial [%]
0	22 Jan 2018	02 Feb 2018	0	<0.010
			100	<0.010
24	23 Jan 2018	02 Feb 2018	0	<0.010	n.a.
			100	<0.010	n.a.
96	26 Jan 2018	02 Feb 2018	0	<0.010	n.a.
			100	<0.010	n.a.

¹     Samples were stored in the freezer (≤ -15°C) until the day of analysis.

²     Percentage of a saturated solution (SS) prepared at a loading rate of 100 mg/L.

n.a.Not applicable.

Table2          
Incidence of Mortality and Total Mortality during the Limit Test

C.I. Pigment Red 63:1, %SS at 100 mg/L	Initial Number of fish	Cumulativemortality					Total Mortality (%)
		2½h	24h	48h	72h	96h
Control	7	0	0	0	0	0	0
100	7	0	0	0	0	0	0

Validity criteria fulfilled:

yes

Remarks:

See "Overall remarks" for details on the acceptability criteria

Conclusions:

An acute fish study was performed with Pigment Red 63:1 according to OECD guideline 203 and GLP principles. Actual test concentrations could not be measured during the test. Therefore, the LOQ of 0.010 mg/L was considered to be above the maximum solubility limit of Pigment Red 63:1 in test medium. Due to the low solubility of C.I. Pigment Red 63:1 in water, concentration levels that might be lethal to carp could not be reached. Therefore, the LC50 was considered to be above the maximum solubility of Pigment Red 63:1 in test medium at all time points during the 96h test period.

Executive summary:

In a 96 -h OECD 203 study according to GLP principles, the acute toxicity of the test substance to carp (Cyprinus carpio) was investigated. A final test was performed based on the results of a preceding range-finding test. Seven carp per test group were exposed to an untreated control and to a saturated solution prepared at 100 mg/L. No mortality or clinical effects were observed at both the SS and the control during the test period. Samples taken from the SS were analysed and showed that the measured concentrations were below the limit of quantification (LOQ was 0.010 mg/L) throughout the test period. Due to the low solubility of the test substance, the LC50 is considered to be above the maximum solubility of the test substance in test medium. The study is considered to be valid.

Description of key information

An acute fish study was performed according to OECD guideline 203 and GLP principles. The acute toxicity of Pigment Red 63:1 to carp (Cyprinus carpio) was found to be above the maximum solubility of the test substance in test medium.

Additional information

In a 96 -h OECD 203 study according to GLP principles, the acute toxicity of the test substance to carp (Cyprinuscarpio) was investigated. A final test was performed based on the results of a preceding range-finding test. Seven carp per test group were exposed to an untreated control and to a saturated solution prepared at 100 mg/L. No mortality or clinical effects were observed at both the SS and the control during the test period. Samples taken from the SS were analysed and showed that the measured concentrations were below the limit of quantification (LOQ was 0.010 mg/L) throughout the test period. Due to the low solubility of the test substance, the LC50 is considered to be above the maximum solubility of the test substance in test medium.The study is considered to be valid.