3,6-bis(ethylamino)-9-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthylium chloride

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:

23 April 1990 to 27 April 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: DIN 38 412 using a static procedure

Version / remarks:

“Testverfahren Mit Wasserorganismen (Gruppe L). Allgemeine Hinweise Zur Planung, Durchfuehrung Und Auswertung Biologischer Testverfahren (L1)” Und "Bestimmung Der Wirkung Von Wasserinhaltsstoffen Auf Fische - Fischtest (L15)”, June 1982

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

only four dose levels were examined

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: In the form of an aqueous solution (1.0 g/100 mL) the product was added to the test water

Test organisms (species):

Leuciscus idus

Details on test organisms:

TEST ORGANISM
- Common name: Golden Orfe
- Length at study initiation: 8.4 cm (range: 7.5 - 9.3 cm)
- Weight at study initiation: 6.0 g (range: 4.4 - 7.6 g)
- Corpulence factor (K) of the batch: 1.0 (k is calculated from the weight (g) and the length (cm) measured from the tip of the mouth to the distal end of the caudal fin according to the formula K = 100*W/L**3).

ACCLIMATION
- Acclimation period: About 5 months in the laboratory, adapted to test water and test temperature for 3 days.
- Acclimation conditions: The fish were kept in a flow-through tank in tap water cleaned by active carbon and aerated with oil-free air; photoperiod 16 hours of light and 8 hours of darkness; total hardness : about 2.5 mmol/; acid capacity about 5.5 mmol/L; oxygen content > 60 % of maximum saturation; pH about 8.0; and temperature 18 to 22 °C.
- Amount of food during acclimation: ad libitum
- Health during acclimation (any mortality observed): Mortality during the last 2 weeks of housing was 0 %; mortality during the adaptation period was 0%
- Medical treatment: Fish were treated twice with 0.05 mg/L malachite green chloride and once with 10 mg/L tetracycline hydrochloride

FEEDING DURING TEST: No
- Time schedule: Food was withdrawn 1 day before the test and was not provided throughout the exposure period.

Test type:

static

Water media type:

freshwater

Remarks:

Reconstituted freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

2.5 mmol/L

Test temperature:

19 - 20 °C

pH:

About 8.0

Dissolved oxygen:

7.8 - 8.7 mg/L

Conductivity:

max. 10 micro mho

Nominal and measured concentrations:

- Nominal concentrations: 1.0, 2.15, 4.64 and 10 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: All glass aquarium (30 x 22 x 24 cm)
- Material, size, headspace, fill volume: 10 L
- Aeration: Continuous aeration with oil-free air
- No. of organisms per vessel: 10 fish per vessel
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 6.0 g fish/L test water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted freshwater according to DIN 38 412, part 11, October 1982, preparation from fully demineralised tap water. The re-salting of test water is carried out by addition of 294.0 mg/L CaCl2.2H2O, 123.3 mg/L MgSO4.7H2O, 63.0 mg/L NaHCO3 and 5.5 mg/L KCl.
- Ca/Mg ratio: 4:1
- Na/K ratio: 10:1

OTHER TEST CONDITIONS
- Adjustment of pH: Not specified

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Determination of mortality and symptoms was carried out after 1, 4, 24, 48, 72 and 96 hours.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca.2.2
- Range finding study: Yes
- Results used to determine the conditions for the definitive study: Yes. 100 % mortality was seen at a concentration of 10 mg/L within 72 hours.

Reference substance (positive control):

yes

Remarks:

Chloroacetamide

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

9.54 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

7.49 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

6.85 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

2.15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

There were no mortalities in the 1.00 and 2.15 mg/L dose groups. In the 4.64 dose group, one fish was observed to be dead after 24 hours and a second fish died after 48 hours. In the highest dose group, 5 fish had died after 48 hours, 7 after 72 hours and 8 after 96 hours.
An observation of a narcotic-like state was made for the fish exposed to 10 mg/L after 24 hours. At 48 and 72 hours, apathy was observed in fish at the 4.64 and 10 mg/L dose levels, and was also observed at 96 hours in the top dose only.

Results with reference substance (positive control):

- Results with reference substance valid? Yes, the lethal concentration corresponds to the normal sensitivity.
- LC50: 48 H value ca. 31 mg/L

Reported statistics and error estimates:

Statistical analysis was carried out using probit analysis according to the method of Finney.

Sublethal observations / clinical signs:

Table 1: Summary of Mortality

Nominal Concentration (mg/L)	Number of Fish	Number of dead fish
		1 hour	4 hours	24 hours	48 hours	72 hours	96 hours
0.00	10	0	0	0	0	0	0
1.00	10	0	0	0	0	0	0
2.15	10	0	0	0	0	0	0
4.64	10	0	0	1	2	2	2
10.00	10	0	0	0	5	7	8

Table 2: Summary of Symptoms

Nominal Concentration (mg/L)	Number of Fish	Symptoms observed
		1 hour	4 hours	24 hours	48 hours	72 hours	96 hours
0.00	10	-	-	-	-	-	-
1.00	10	-	-	-	-	-	-
2.15	10	-	-	-	-	-	-
4.64	10	-	-	-	(A)	(A)	-
10.00	10	-	-	N	A	A	A

Explanation of symptoms:

A = Apathy

N = Narcotic-like state

- = No symptoms observed

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the study, the 96 hour LC50 was determined to be 6.85 mg/L and the NOEC was 2.15 mg/L.

Executive summary:

The acute toxicity of the test material to golden orfe (Leuciscus idus) was investigated in accordance with the guideline DIN 38 412 using methodology equivalent to that of the standardised guideline OECD 203.

Following a preliminary study, fish were exposed to the test material at nominal concentrations of 0, 1.0, 2.15, 4.64 and 10 mg/L under static conditions. The test material was dissolved in test water and fish were exposed in groups of ten per dose level for 96 hours. Determination of mortality and symptoms was carried out after 1, 4, 24, 48, 72 and 96 hours. 

There were no mortalities in the 1.00 and 2.15 mg/L dose groups. In the 4.64 dose group, one fish was observed to be dead after 24 hours and a second fish died after 48 hours. In the highest dose group, 5 fish had died after 48 hours, 7 after 72 hours and 8 after 96 hours.

An observation of a narcotic-like state was made for the fish exposed to 10 mg/L after 24 hours. At 48 and 72 hours, apathy was observed in fish at the 4.64 and 10 mg/L dose levels, and was also observed at 96 hours in the top dose only.

Under the conditions of the study, the 96 hour LC50 was determined to be 6.85 mg/L and the NOEC was 2.15 mg/L.

Description of key information

Under the conditions of the study, the 96 hour LC50 was determined to be 6.85 mg/L and the NOEC was 2.15 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

6.85 mg/L

Additional information

The acute toxicity of the test material to golden orfe (Leuciscus idus) was investigated in accordance with the guideline DIN 38 412 using methodology equivalent to that of the standardised guideline OECD 203. The study was assigned a reliability score of 2 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

Following a preliminary study, fish were exposed to the test material at nominal concentrations of 0, 1.0, 2.15, 4.64 and 10 mg/L under static conditions. The test material was dissolved in test water and fish were exposed in groups of ten per dose level for 96 hours. Determination of mortality and symptoms was carried out after 1, 4, 24, 48, 72 and 96 hours. 

There were no mortalities in the 1.00 and 2.15 mg/L dose groups. In the 4.64 dose group, one fish was observed to be dead after 24 hours and a second fish died after 48 hours. In the highest dose group, 5 fish had died after 48 hours, 7 after 72 hours and 8 after 96 hours.

An observation of a narcotic-like state was made for the fish exposed to 10 mg/L after 24 hours. At 48 and 72 hours, apathy was observed in fish at the 4.64 and 10 mg/L dose levels, and was also observed at 96 hours in the top dose only.

Under the conditions of the study, the 96 hour LC50 was determined to be 6.85 mg/L and the NOEC was 2.15 mg/L.