Reaction mass of [5-methyl-2-[[4-(4-methylanilino)-9,10-dioxo-1-anthryl]amino]phenyl]sulfonyloxypotassium and [5-methyl-2-[[4-(4-methyl-2-potassiooxysulfonyl-anilino)-9,10-dioxo-1-anthryl]amino]phenyl]sulfonyloxypotassium

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:

From 18 to 22 November (preliminary); from 2 to 6 December, 1996 (main)

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

adopted 17 July, 1992

GLP compliance:

yes

Analytical monitoring:

no

Test organisms (species):

Poecilia reticulata

Details on test organisms:

TEST ORGANISM
- Common name: Guppy
- Length at study initiation: 1 - 3 cm
- Fasting before test: fish were last fed 24 hours before the start of the definitive test.

BREEDING
- Breeding: parental and young fish were held in two 250 l glass aquaria.
- Illumination: 14 h a day
- temperature: 23 ± 2 °C
- Medium: continuosly aerated and filtered tap water from Schlleren.The bottom of the aquaria was covered with gravel (less than 1.5 cm). Every month, one third of the total volume of the aquaria was replaced by fresh tap water.
- Feeding frequency during acclimation: Sera micropam, Sera vipan, daily. The


ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions: fish were held in the batch used for the test.
- Health: mortality was less than 0.1 %.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

Test vessels: 21 - 22
Control:: 21 - 22

pH:

Test vessels: 8.0 - 8.8
Control: 8.0 - 8.6

Dissolved oxygen:

Test vessels: 6.2 - 8.6 mg/l
Control: 6.3 - 8.3 mg/l

Nominal and measured concentrations:

4, 8, 16, 32 and 64 mg/l, nominal

Details on test conditions:

TEST SYSTEM
- Test vessel: 1 l aquaria, all-glass.
- Aeration: aerated.
- No. of organisms per concentration: 7 fish.
- No. of vessels per concentration: one replicate.
- No. of vessels per control: one replicate.

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: tap water from Zürich-Schkkeren.

OTHER TEST CONDITIONS
- Photoperiod: 14 hours a day, supplied by overhead white fluorescent tubes.

EFFECT MEASURED
Fish were considered to be dead when they showed no reaction after touching the caudal fin or no visible breathing movements.
Observations: were made after 2, 24, 48, 72 and 96 hours of exposure.

EFFECT PARAMETERS MEASURED
Oxygen, pH, temperature: prior to the addition of fish to the test medium and after 2, 24, 48, 72 and 96 hours of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0
- Range-finding: prior to the definitive test a range-finding test with concentrations of 1, 10 and 100 mg/l and 3 fish per test vessel was conducted.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

14.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The no-observed effect concentration (NOEC) was 8 mg/l since the one fish that died was considered not to represent a significant effect as defined by the validity criteria of the test.
100 % mortality (LC100) was seen at the nominal concentration of 32 mg/l.
The following mortalities (of a total of 7 fish) were observed in the definitive test at the different test substance concentrations after 96 hours of exposure: 64 mg/l (7), 32 rng/l (7),16 mg/l (4), 8 mg/l (1) and g mg/l (1).Non mortality was observed in the control after 96 h of expesure.
Other toxic effects than mortality, e.g. loss of oordination, hypoactivity and swimming on the back, were not observed.

RANGE FINDING
100 % mortality was observed at 100 mg/l after 4 hours of exposure. 33 % mortality was observed at 10 mg/l after 48 hours of exposure. No mortality was observed at 1 mg/l and in the control after 96 hours of exposure.

Sublethal observations / clinical signs:

Incidence of dead fish observed at different points in time after start of exposure.

Nominal concentration (mg/l)	Number of dead fish observed at different points in time after start of exposure
	2 hrs	24 hrs	48 hrs	72 hrs	96 hrs
Control 0	0	0	0	0	0
Test item 4	0	1	0	0	0
Test item 8	0	1	0	0	0
Test item 16	0	1	2	1	0
Test item 32	3	4	-a)	-a)	-a)
Test item 64	6	1	-a)	-a)	-a)

a) All fish died

Conclusions:

LC50 (96h): 14.3 mg/l (nominal)

Executive summary:

The acute lethal toxicity (LC50) of test item to Poecilla reticulata (Guppy) was investigated under static exposure conditions over a period of 96 hours. 7 fish per concentration were exposed to the test substance. The nominal concentrations at test item as selected by a range finding test were 4, 8, 16, 32 and 64 mg/l, respectivety. The individual test concentrations were prepared by dilution of a stock solution. No chemical analyses of the test concentrations were conducted.

The following mortalities (of a total of 7 fish) were observed in the definitive test at the different test substance concentration after 96 h of exposure: 64 mg/l (7), 32 rng/l (7),16 mg/l (4), 8 mg/l (1) and g mg/l (1). Non mortality was observed in the control after 96 h of exposure.

Other toxic effects than mortality, e.g. loss of coordination, hypoactivity and swimming on the bank, were not observed.

Based on these data the noninal lethal concentration (LC50) of test item to Poecilla reticulata was calculated to be 14.3 mg/l.

The no-observed effect concentration (NOEC) was 8 mg/l since the one fish that died was considered not to represent a significant effect as defined by the validity criteria of the test.

100 % mortality (LC100) was seen at the nominal concentration of 32 mg/l.

Conclusion

LC50 (96h): 14.3 mg/l (nominal)

Description of key information

LC50 (96h): 14.3 mg/l (nominal)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

14.3 mg/L

Additional information

The acute lethal toxicity of test item to Poecilla reticulata was investigated under static exposure conditions over a period of 96 hours. The individual test concentrations were prepared by dilution of a stock solution. No chemical analyses of the test concentrations were conducted. The noninal lethal concentration (LC50) was calculated to be 14.3 mg/l; the no-observed effect concentration (NOEC) was 8 mg/l, while the 100 % mortality (LC100) was seen at the nominal concentration of 32 mg/l. Other toxic effects than mortality, e.g. loss of coordination, hypoactivity and swimming on the bank, were not observed.

An old test performed on the substance under registration is also available and indicate a LC50 value at 48 hours of 2 mg/l. The LC0 was indicated to be 1 mg/l, while the LC100 was idicated at 5 mg/l (reference, 1977). There are no details on testing method and procedures followed; only the sheet of results is available, thus reliability cannot be assigned and the data has not been used for classification.

Data on the structural analogous Similar Substance 02 have also been taken into consideration, in order to clarify the potential toxicity of the substance to fish; the read across approach can be considered as appropriate and suitable to assess the property under investigation (details about the approach are reported into the IUCLID section 13).

The acute lethal toxicity (LC50) of Similar Substance 02 to rainbow trout was investigated under semi-static exposure conditions, over a period of 96 hours. The nominal concentration for the definitive test was established on the basis of the range-finding study outcomes: the substance was tested at 1.0, 10 and 100 mg/l, under static conditions; 100 % of mortality was recorded at the highested tested concentration. Thus, the limit test was conducted at 10 mg/l. During the definitive test, no deaths occurred either in the test vessels, neither in the control one.

REFERENCE

See attachment