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

Long-term toxicity to fish

Administrative data

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

July 9 2002 - August 10 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted to current recognised methodology

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)

Deviations:

yes

Principles of method if other than guideline:

The temperature fluctuation in the course of the study exceeded 1.5°C and did not remain in the temperature range from 23-27°C. The results obtained for the control (hatching success 88.3%; post hatch success 88.7%) demonstrate that this slight deviation from the demands of the guideline does not affect the validity of the study.

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): Procion Karminrot H112239
- Molecular formula (if other than submission substance): C53 H28 Cl2 N14 O26 S8 Na8
- Molecular weight (if other than submission substance): 1785.0 g/mol (according to data of the sponsor)
- Substance type: Organic
- Physical state: Solid
- Analytical purity: 63.8 % (according to data of the sponsor)
- Impurities (identity and concentrations): None identified
- Lot/batch No.: 5K/Brunsbuttel
- Stability under test conditions: Verified by chemical analysis (HPLC) during the study.

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:

- Concentrations: 1.0, 3.16 and 10 mg/l
- Sampling method: Direct sampling
- Sample storage conditions before analysis: None; analysis conducted post sampling.

Test solutions

Vehicle:

no

Details on test solutions:

Identity and concentration of auxiliary solvent for dispersal: None; a stock solution was prepared for the required concentrations.

Test organisms

Test organisms (species):

Pimephales promelas

Details on test organisms:

Type: Fathead minnow (Pimephales promelas RAFINESQUE)
Source: laboratory brood (strain: Bio International, Netherlands)
Date of Birth: March 9,2001
Acclimatisation: Stock acclimatised since January 31, 2002 to the test conditions.
Temperature: 23 - 27 °C
Dissolved Oxygen: >5mg/l
Feeding: Tetramin fish food, ground, 2 % of body weight per day, applicated in 3 portions daily (10.00 a.m., 13.00 a.m., 4.00 p.m.) Artemia salrna, Daphnids or Lavae of Chironomus. ripariits (frozen) ad libitum, daily. On weekends feeding only with Tetra Min fish food.
Mortalities during acclimatisation period: < 5 %

Medication: none

Holding and dilution water: Reconstituted water prepared according to the recommendations of ISO 7346. This freshly prepared standard dilution water is used for both, the maintenance of the test animals under flow-through conditions and the preparation of stock and test solutions of the test substance.

Study design

Test type:

flow-through

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

32 d

Post exposure observation period:

None

Test conditions

Hardness:

The total hardness of the dilution water, measured 5 times during the test, ranged from 9.8°dH- 125°dH(= 174.9 mg/l CaCO3-223.1 mg/l CaCO3).

Test temperature:

23 - 27°C

pH:

7.5-7.9

Dissolved oxygen:

7.3 - 8.4 mg/l

Salinity:

Not applicable

Nominal and measured concentrations:

1.0, 3.16 and 10 mg/l (nominal)

Details on test conditions:

Pretreatment of the test substance: A stock solution was prepared to give the desired series of test concentrations.

Exposure conditions:

Test Vessels: glass vessels, diameter 14 cm, height 7.5 cm glass aquaria holding 51 of test media

Experimental design:
3 test concentrations plus control
60 fertilised eggs per test concentration and control evenly distributed into two replicates
flow-through system

Method of initiation: fertilised eggs (16 - 256 cell stage) were placed hi prepared media

Photoperiod: 16h light:8h dark

Temperature: mean +/-1.5 °C (in the temperature range 23 - 27 °C)

Aeration: gentle aeration via narrow glass tubes after hatching of larvae (start of aeration: July 15,2002)

Test concentration/s (nominal): 1.0, 3.16 and 10 mg/l

Method of administration: stock solution

Medium renewal: continuously

Duration of exposure: 32 days

Criteria of effects:

- Hatching success;
- Mortality,
- Body abnormalities;
- Type and incidence of behavioural abnormalities;
- Length of juvenile fish at the end of the test period;
- Fresh-weight of juvenile fish at the end of the test period

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

32 d

Dose descriptor:

NOEC

Effect conc.:

3.61 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Total Survival

Effect concentrations 2

Duration:

32 d

Dose descriptor:

LOEC

Effect conc.:

10.43 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Total survival

Effect concentrations 3

Duration:

32 d

Dose descriptor:

NOEC

Effect conc.:

>= 10.34 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Hatching success, post hatch survival, length

Effect concentrations 4

Duration:

32 d

Dose descriptor:

LOEC

Effect conc.:

> 10.34 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Hatching success, post hatch survival, length

Details on results:

Signs of toxicity: None visible

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

The statistical analysis of the results was performed using the software „Toxrat (ToxRat Solutions GmbH, Alsdorf, Germany). The following parameters were statistically evaluated: post hatch survival (larval survival) after 32 d, total survival after 32d, weight of fish after 32d, length of fish after 32d.

As no dose response relationship is obvious for the parameter ..hatching success", and additionally the mean hatching success of the highest test substance concentration was only 5% below the observed value of the control, no statistical determination of the NOEC/LOEC'has been performed. As all hatching rates observed exceeded the demands of the OECD Guideline 210 the highest test substance concentration of nominally 10.0 mg/l was considered to be the No Observed Effect Concentration (NOEC)

Any other information on results incl. tables

The temperature fluctuation in the course of the study exceeded 1.5°C and did not remain in the temperature range from 23-27°C. The results obtained for the control (hatching success 88.3%; post hatch success 88.7%) demonstrate that this slight deviation from the demands of the guideline does not affect the validity of the study,

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The recognised NOEC is 3.61 mg/l after 32 days based on survival & weight of juveniles

Executive summary:

Based on different biological effects observed and endpoints assessed, the following results were obtained at the end of the 32 day exposure period of the Fish. Early-life Stage Toxicity Test:

 

Results [mg/l]:

NOEC(Total survival)                                              3.61 mg/l.

LOEC (Total survival)                                             10.43 mg/l

 

NOEC(Hatching success, post hatch survival, length)    ≥ 10.43 mg/l.

LOEC(Hatching success, post hatch survival, length)     > 10.43 mg/l                                  

 

All results are expressed in terms of mean measured concentrations during exposure period. Measured concentrations ranged from 69.0% -101% of nominal values of Procion Karminrot H-EXL - H112339 during acclimatisation phase and from 90.7% -152.8% of nominal values of Procion Karminrot H-BXL-H 112339 during exposure period.

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