Benzoic acid, 2-hydroxy-5-[[4-[[4-[[8-hydroxy-7-[[4-[(8-hydroxy-3,6-disulfo-1-naphthalenyl)azo]-2-methoxy-5-methylphenyl]azo]-3,6-disulfo-1-naphthalenyl]amino]-6-(phenylamino)-1,3,5-triazin-2-yl]amino]phenyl]azo]-, pentasodium salt

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From March 14 to May 5, 2007

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No analytical monitoring

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1984

GLP compliance:

no

Remarks:

in accordance with ISO/IEC 17025

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

Breeding system
- test organism: Daphnia magna (Straus 1820)
- breeding: parental and young Daphnia were held in 250 l glass aquaria
- illumination: 16 h per day
- temperature: 20 ± 2 °C
- medium: same as used for the test
- feeding: once a day, except weekends
- food type: 15 g sera micropan homogenised in 1 l deionised water

Acclimation
- acclimation period: 4 h
- acclimation conditions: yes

Control of sensitivity: periodically conducted with reference substance, potassium dichromate

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20 ± 0.5 °C

pH:

7.2 - 7.7

Dissolved oxygen:

6.0 - 6.2 mg/l

Nominal and measured concentrations:

Nominal concentrations: 1, 10, 100 mg/l
Measured concentrations not available

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 ml flasks, all-glass, with 50 ml of test medium
- Aeration: not aerated
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water according to ISO 6341 (1996). Final test medium contained 25 ml/l of each of the following stock solutions:
CaCl2 × 2H2O, 11.76 g/l
MgSO4 × 7H2O, 4.93 g/l
NaHCO3, 2.59 g/l
KCl, 0.23 g/l
- Alkalinity: 0.8 mmol/l
- Ca:Mg ratio: 4:1
- Na:K ratio: 10:1
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 16 hours a day, supplied by overhaed white fluorescent tubes

EFFECT PARAMETERS MEASURED: immobilisation, at 24 and 48 h after start of exposure

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Immobilisation of daphnia

nominal concentration (mg/l)	code	no. of individuals per test vessel	immobile daphnia (no./vessel) 24 h	immobile daphnia (no./vessel) 48 h	% immobile daphnia after 24 h	% immobile daphnia after 48 h
0 (control)	A	20	0	0	0	0
	B	20	0	1	0	5
	C	20	0	0	0	0
1	A	20	0	0	0	0
	B	20	0	1	0	5
10	A	20	0	2	0	10
	B	20	0	1	0	5
100	A	20	0	3	0	15
	B	20	0	2	0	10

nominal concentration (mg/l)	code	pH values - 0 h	pH values - 48 h	oxygen con. (mg/l) 0 h	oxygen conc. (mg/l) 48 h
0 (control)	A	7.3	7.5	6.0	6.1
	B
	C
1	A	7.3	7.7	6.1	6.1
	B
10	A	7.3	7.6	6.0	6.2
	B
100	A	7.2	7.7	6.1	6.1
	B

Validity criteria fulfilled:

yes

Conclusions:

EC50 (48h) > 100 mg/l.

Executive summary:

Method

Acute toxicity to daphnia magna was investigated under static exposure conditions over a period of 48 h. A screening test with nominal concentrations of 100, 10 and 1 mg/l was performed. Each concentration was tested in 2 replicates, each containing 20 organisms. The final test concentration was prepared by dilution of a stock solution.

No analytical verification was conducted and nominal concentrations were used, assuming the compound is stable in water over 48 h.

Results

At concentrations of 1 and 10 mg/l as well as in the control, no significant effects (> 10 % immobilisation) were noted after 24 and 48 h of exposure. At 100 mg/, 12.5 % effects were noted after 48 h.

Accordingly, EC50 values at 24 and 48 h were estimated to be > 100 mg/l, while EC0 was between 10 and 100 mg/l at 48 h.

Description of key information

EC50 (48h) > 100 mg/l

EC0 (48h) = 10 -100 mg/l

Additional information

The substance was tested for acute toxicity on aquatic invertebrates, according to the OECD guideline 202.

During the test, no analytical determination of concentration was done, as solutions were assumed to be sufficiently stable within the 48 -hour exposure period. Concentrations of 1, 10 and 100 mg/l were tested. No significant immobilisation was noted in controls as well as at 1 and 10 mg/l, while the highest oncentration showed 12.5 % effects after 48 h.

A further study (1982) was available using a test sample with highpurity. An EC50 value of 209 mg/l was reported, but no details on procedure and result were available. Therefore, this data was only taken as a qualitative indication of low toxicity to daphnia.