Disodium 2-(11-oxido-3-oxo-3H-dibenzo[c,h]xanthen-7-yl)benzoate

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:

supporting study

Study period:

2010-03-15 to 2010-03-24

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992-07-17

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008-05-30

Deviations:

no

GLP compliance:

not specified

Remarks:

The study was conducted in accordance with ISO/IEC 17025.

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature, protected from light.

Analytical monitoring:

no

Remarks:

No analyses of test concentrations were conducted. For the evaluation of the test the nominal concentration of the test substance in the test media was used.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: As a stock solution the nominal test concentrations of 100 mg/L was prepared. Test concentrations were prepared by dilution of this stock solution.
- Controls: Untreated test medium
- Evidence of undissolved material: No

Test organisms (species):

Poecilia reticulata

Details on test organisms:

TEST ORGANISM
- Common name: Guppy
- Length at study initiation: 1 - 3 cm
- Method of breeding: Population of parental and young fish was held in either 200 L or 40 L glass aquaria.
- Maintenance of the brood fish:
Illumination: 16 h per day
Temperature: 21 - 25 °C (thermostatic heating element)
Medium: Continuously aerated and filtered tap water. The bottom of the aquaria was covered with gravel (<1.5 cm).
Feed: Sera micropan, Sera vipan (Sera apanristic GmbH, D-52525 Heinsberg, Germany), daily. The fish were last fed 72 h before the start of the test series.
Validity: During 7 days before the test series, mortality in the batch of which fish were used for the test was less than 5 %.

ACCLIMATION
- Acclimation conditions: same as test
- Health during acclimation: During 7 days before the range finding test, no mortality in the batch of fish, which have been bred in the laboratory, was observed.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

36.6 - 37.5 °f H

Test temperature:

21 - 24 °C,

pH:

7.2 - 7.4

Dissolved oxygen:

Start:
Control: 8.0 mg/L O2
1.0 mg/L: 7.9 mg/L O2
10 mg/L: 8.2 mg/L O2
100 mg/L : 8.0 mg/L O2

End:
Control: 7.0 mg/L O2
1.0 mg/L: 6.9 mg/L O2
10 mg/L: 7.0 mg/L O2
100 mg/L: 6.6 mg/L O2

Conductivity:

691 - 731 µS/cm

Nominal and measured concentrations:

Nominal: 0, 1.0, 10, and 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: aquaria
- Type: open
- Material, fill volume: glass, 1 L
- Aeration: No
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Tap water
- Chlorine: 11.5 - 19.0 mg/L
- Culture medium different from test medium: No


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

EFFECT PARAMETERS MEASURED: Observations of the fish were made after 2, 24, 48, 72 and 96 h of exposure. Mortality of the fish was recorded at the same time intervals. Dead fish were removed immediately from the test medium and weight and length recorded.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10

Reference substance (positive control):

no

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)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 6.3 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: No
- Observations on body length and weight: No
- Other biological observations: No
- Mortality of control: 1 fish died in the blank control after 96 h of exposure.
- Other adverse effects control: No
- Abnormal responses: No
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

Sublethal observations / clinical signs:

Table 1: Incidence of Mortality

Nominal concentration (mg/L)	Number of dead fish observed at different points in time after start of exposure
	2 h	24 h	48 h	72 h	96 h
Control	0	0	0	0	1
1.0	0	2	0	0	1
10.0	0	0	0	0	0
100.0	0	0	0	1	0

Table 2: Data of surviving fish after termination of the test

Nominal concentration (mg/L)
1.0	Length (mm)	19	25	17	24	-	-	-
	Weight (mg)	39	81	35	116	-	-	-
10.0	Length (mm)	20	20	22	18	17	21	20
	Weight (mg)	49	58	96	44	32	63	55
100.0	Length (mm)	18	22	20	19	18	19	-
	Weight (mg)	47	86	48	51	42	45	-

Validity criteria fulfilled:

yes

Conclusions:

In an 96 h acute toxicity test according to OECD Guideline 203 in Poecilia reticulata (Guppy), a LC50 of > 100 mg/L was determined.

Executive summary:

In an acute toxicity test according to OECD Guideline 203, the acute lethal toxicity of an aqueous formulation containing 6.3 % test item to Poecilia reticulata (Guppy) was investigated under static exposure conditions over a period of 96 h. The nominal concentrations of Security Indicator BX Liquid were 1,10 and 100 mg/L, respectively. Therefore, this study represents a screening test. The individual test concentrations were prepared by dilution of a stock solution. No analytical verification of the test substance concentration was conducted. The evaluation of the test was based on the nominal concentrations of the test material, assuming the test compound to be stable in water over 96 h. After 96 h of exposure a total of 3 and 1 of seven fish were dead at 1 and 100 mg/L. No mortality was observed at 10 mg/L. 1 fish died in the blank control after 96 h of exposure. Toxic effects other than mortality, e.g. loss of coordination, hypo- or hyperactivity and swimming on the back, were not observed. The no-observed-effect concentration (NOEC, LC0) was >100 mg/L since the one fish that died is considered not to represent a significant effect as defined by the validity criteria of the test. Since minor effects were observed at 100 mg/L, the three dead fish observed at 1 mg/L were rated to be not a significant effect. 100 % mortality (LC100) could not be detected. Based on the binominal theory, the absence of mortality provides at least 99 % confidence that the LC50 is greater than the highest applied nominal test concentration of 100 mg/L (LC50 recalculated for active ingredient = 6.3 mg/L).

Description of key information

In an 96 h acute toxicity test according to OECD Guideline 203 in Poecilia reticulata (Guppy), using an aqueous formulation containing 6.3 % of the test item, an LC50 of > 100 mg/L was determined (LC50 recalculated for active ingredient = 6.3 mg/L).

Additional information

In an acute toxicity test according to OECD Guideline 203, the acute lethal toxicity of an aqueous formulation containing 6.3 % test item to Poecilia reticulata (Guppy) was investigated under static exposure conditions over a period of 96 h. The nominal concentrations of the formulation were 1, 10 and 100 mg/L, respectively. Therefore, this study represents a screening test. The individual test concentrations were prepared by dilution of a stock solution. No analytical verification of the test substance concentration was conducted. The evaluation of the test was based on the nominal concentrations of the test material, assuming the test compound to be stable in water over 96 h. After 96 h of exposure a total of 3 and 1 of seven fish were dead at 1 and 100 mg/L, respectively. No mortality was observed at 10 mg/L. 1 fish died in the blank control after 96 h of exposure. Toxic effects other than mortality, e.g. loss of coordination, hypo- or hyperactivity and swimming on the back, were not observed. The no-observed-effect concentration (NOEC, LC0) was >100 mg/L since the one fish that died is considered not to represent a significant effect as defined by the validity criteria of the test. Since minor effects were observed at 100 mg/L, the three dead fish observed at 1 mg/L were rated to be not a substance related effect. 100 % mortality (LC100) could not be detected. Based on the binominal theory, the absence of mortality provides at least 99 % confidence that the LC50 of the formulation is greater than the highest applied nominal test concentration of 100 mg/L (LC50 recalculated for active ingredient = 6.3 mg/L).