Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 December 1991 to 13 December 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to EU recognised test guideline in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

84/449/EEC

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

EC Number:

413-550-5

EC Name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Cas Number:

142068-96-0

Molecular formula:

Hill formula: C53H28Cl2N14Na8O26S8

IUPAC Name:

octasodium 5-[(4-chloro-6-{[4-({[4-chloro-6-({7-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-8-hydroxy-3,6-disulfonatonaphthalen-1-yl}amino)-1,3,5-triazin-2-yl]amino}methyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-3-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-4-hydroxynaphthalene-2,7-disulfonate

Details on test material:

- Name of test material (as cited in study report): H112339 (reference as sample V723)
- Physical state: Dark brown powder
- Lot/batch No.: NBY 405/74
- Storage condition of test material: darkness at an ambient temperature

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The concentration of the sample V723 in the test solutions was measured at the start, and end of the 96 hour exposure period.

Test solutions

Vehicle:

no

Details on test solutions:

The test solution was prepared by dispensing an appropriate weight of sample in the 15 litres test volume of water in the test vessel.
The following nominal exposure concentration was employed: 112 mg V723/litre together with a freshwater control (this is to account for the water impurity in the test sample)

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

The test organism was the rainbow trout {Salmo gairdneri), a sensitive freshwater fish. The fish were obtained from Upwey Trout Farm, Upwey, Dorset on 8 October 1991 and held in stock tanks of the ICI Group Environmental Laboratory.

Immediately prior to the test the fish were acclimatised to the test temperature (15 ± 1°C) for a minimum period of 7 days.

During the holding and acclimation periods the fish were fed appropriate amounts of a commercial fish food.

The fish tested ranged in weight from 0.61 g to 1.24 g with a mean weight of 0.85 g. The range in length was 36 to 43 mm with a mean length of 39 mm (data as measured on the control fish at the end of the test).

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No post exposure observation

Test conditions

Hardness:

52 mg/l CaCO3,

Test temperature:

15°C (± 1)

pH:

7.6 - 7.8

Dissolved oxygen:

9.2 - 9.8 mg/l

Salinity:

Not applicable.

Nominal and measured concentrations:

>112 mg test material / l or >100 mg Substance HI 12339 mg/l

Details on test conditions:

The apparatus used in this study were glass vessels of nominal working capacity 15 hues. These vessels were aerated during the period of the test, and one vessel was used for the exposure concentration and one for the control.
At the start of the test the fish were transferred into the test vessels containing the test solution prepared as noted under 3.2 above. Ten fish were used for the test concentration and the control giving a fish loading of 0.57 g/1 based on the volume of 15 1 of solution used in the test and the mean weight of the fish .

The temperature was maintained at 15 ± 1°C and the light regime was fluorescent lighting for 16 hr and darkness for 8 hr.

The trout were not fed for 48 hours before and during the course of the test.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 4

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 5

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

As recorded no fish died during the study and the 24, 48, 72 and 96 hr LC50 values must all be >112 mg V723/1 or >100 mg Substance HI 12339 mg/1. Similarly no symptoms of toxicity were observed and the 96 hour NOEC (no observed effect concentration) is therefore > 100 mg/1 Substance HI 12339.

The 24, 48, 72 and 96 hour median lethal concentrations of Substance HI 12339 to the freshwater fish rainbow trout (Salmo gairdneri) were determined under static test conditions at a temperature of 15 ± 1°C.
The median lethal concentration (LC50) is defined as the concentration, calculated from the data obtained, resulting in death of 50% of the fish in the time period specified.
The values obtained were:
24 hour LC50 >100 mg/litre
48 hour LC50 >100 mg/litre
72 hour LC50 >100 mg/litre
96 hour LC50 >100 mg/litre

No mortalities or symptoms of toxicity were observed over the 96 hour period at the nominal test concentration of 100 mg Substance H112339/litre and the 96 hour NOEC (no observed effect concentration) is therefore >100 mg/1.
The values obtained were based on the nominal concentrations of Substance HI 12339. The mean measured concentration of the test material was 95% of the nominal value.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

None

Any other information on results incl. tables

Sublethal observations / clinical signs:

Observations of mortalities

 

Time	Nominal concen- tration  (mg/1)	Number tested	Number dead	% dead^1
(hours)
24	Control	10	0	0
	112	10	0	0
48	Control	10	0	0
	112	10	0	0
72	Control	10	0	0
	112	10	0	0
96	Control	10	0	0
	112	10	0	0

'No symptoms of toxicity were observed in any of the test fish at the daily observation times.

 

Analytical results

Nominal times (hours)	Nominal concen­tration V723 (mg/1)	Analysed concen­tration V723 (mg/1)	% nominal
0	112	106.7*	95
96	112	106.7*	95
	Mean	106.7	95

 

*Mean of 3 determinations

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 24, 48, 72 and 96 hour median lethal concentrations of Substance HI 12339 to the freshwater fish rainbow trout (Salmo gairdneri) were determined under static test conditions at a temperature of 15 ± 1°C.
The median lethal concentration (LC50) is defined as the concentration, calculated from the data obtained, resulting in death of 50% of the fish in the time period specified.
The values obtained were:
24 hour LC50 >100 mg/litre
48 hour LC50 >100 mg/litre
72 hour LC50 >100 mg/litre
96 hour LC50 >100 mg/litre

No mortalities or symptoms of toxicity were observed over the 96 hour period at the nominal test concentration of 100 mg Substance H112339/litre and the 96 hour NOEC (no observed effect concentration) is therefore >100 mg/1.

Executive summary:

Study condcuted to EU test guidance 84/449/EEC part C1 in compliance with GLP.

No mortalities or symptoms of toxicity were observed over the 96 hour period at the nominal test concentration of 100 mg/l and the 96 hour NOEC is therefore >100 mg/l. The substance is not classified on the basis of the effects noted.