1-Naphthalenesulfonic acid, 6-diazo-5,6-dihydro-5-oxo-, ester with phenyl(2,3,4-trihydroxyphenyl)methanone

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:

18 Apr - 12 May 1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

04 Apr 1984

Deviations:

yes

Remarks:

Only four test concentrations instead of the recommended five concentrations.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

Directive 84/449/EWG
25 Apr 1984

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: "Letale Wirkung beim Zebrabärbling Brachydanio rerio", Empfehlung des Umweltbundesamtest zur Durchführung der Prüfung nach §4 Abs. 1 Nr. 6 der Verordnung über Anmeldeunterlagen und Prüfnachweise nach dem Chemikaliengesetz Texte 16/84, Umweltbundesamt

Version / remarks:

01 May 1984

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1-Naphthalenesulfonic acid, 6-diazo-5,6-dihydro-5-oxo-, ester with phenyl(2,3,4-trihydroxyphenyl)methanone

EC Number:

270-931-7

EC Name:

1-Naphthalenesulfonic acid, 6-diazo-5,6-dihydro-5-oxo-, ester with phenyl(2,3,4-trihydroxyphenyl)methanone

Cas Number:

68510-93-0

Molecular formula:

all potential esters of C10HH5O3N2SCl and C13H10O4

IUPAC Name:

3-benzoyl-2-hydroxy-6-({[5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalen-1-yl]sulfonyl}oxy)phenyl 5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalene-1-sulfonate; 4-benzoyl-2,3-bis({[5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalen-1-yl]sulfonyl}oxy)phenyl 5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalene-1-sulfonate; 4-benzoyl-2,3-dihydroxyphenyl 5-oxo-6-(λ⁵-diazynylidene)-5,6-dihydronaphthalene-1-sulfonate

Test material form:

solid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 10, 22, 50, and 100 mg/L
- Sampling method: At test begin and after 48 and 96 h, 100 mL were sampled from the center of the tank using a glass screw cap bottle.
- Sample storage conditions before analysis: All analyses were performed on the same day samples were taken.

Test solutions

Vehicle:

yes

Remarks:

Tween 80

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A paste of the test substance was prepared using 0.095 mL Tween 80/L dilution water, which was homogenized with an Ultra-Turrax and ultrasonic bath and added to the test tanks under constant stirring with a glass rod. Then the entire tank content was stirred for ca. 2 h with a KPG-stirrer and glass rod. The test medium presented itself as an orange to red solution or turbid dispersion with substance deposition at the bottom of the tank.
- Controls: Yes, 0 mg/L test substance in test medium (reconstituted water).
- Chemical name of vehicle: Tween 80
- Concentration of vehicle in test medium:0.095 mL Tween 80/L
- Evidence of undissolved material: The test preparation was turbid and substance deposition was observed at the bottom of the test tank.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: West Aquarium, Bad Lauterberg, Germany
- Age at study initiation: ca. 4 months (Hatching date: 25 Dec 1988; Delivery date: 22 Mar 1989).
- Length at study initiation: mean over all test groups = 2.7 cm (SD = 0.23 cm; Min = 2.3, Max = 3.2 cm)
- Maintenance of the brood fish: The fish were held in dilution water for at least 14 d before test start under the following conditions: Temperature: 22 ± 1 °C; Oxygen: ≥ 80% saturation value; Photoperiod: 12 h; Biomass loading: ≤ 1 g fish/L water; Feeding: twice a day, ad libitum; Food type: Tetra Min, Tetra Werke, Melle, Germany

FEEDING DURING TEST
No feeding during test.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Test temperature:

control: 21.8 - 22.9
treatment: 21.8 - 22.9

pH:

control: 7.4 - 8.2
treatment: 7.5 - 8.1

Dissolved oxygen:

control: 6.8 - 9.5
treatment: 5.3 - 8.6

Nominal and measured concentrations:

Control, vehicle control, 10, 22, 50, and 100 mg/L (nominal)
NA, NA, 9.4, 22.3, 50.2, and 95.9 mg/L (mean measured)

Details on test conditions:

TEST SYSTEM
- Test vessel: test tank
- Material, size, headspace, fill volume: Glass test tanks (Length = 30 cm, Width = 22 cm; Height = 22 cm) filled with 10 L test solution.
- Aeration: Before the test the dilution water (reconstituted water) was aerated until oxygen saturation was reached.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 3
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: ≤ 1 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water, according to ISO/DIS 7346/1
- Culture medium different from test medium: Same as test.
- Intervals of water quality measurement: The pH value, oxygen concentration and temperature were measured after 2 - 4, 24, 48, 72, and 96 h. Test item concentrations were measured after 0, 48, and 96 h.

OTHER TEST CONDITIONS
- Photoperiod: 12 h light/12 h dark
- Light intensity: 700 Lux
- Other: The test tanks were placed in a water bath made of Hostalit Z with a transparent Plexiglass screen. The water bath was adjusted to 22 ± 1 °C. No aeration of tanks during test.

EFFECT PARAMETERS MEASURED:
- Mortality: after 2-4, 24, 48, 72, and 96 h
- Aspect & behaviour: after 2-4, 24, 48, 72, and 96 h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Range finding study: Yes.
- Results used to determine the conditions for the definitive study: Yes.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

>= 22 - <= 50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: Behavioural abnormalities were observed at the lowest concentration and higher and after 2-4 h and later. These included: increased activity, braced gills, irregular breathing, disequilibrium, heightened startle response, cramps, lethargy, trembling, spinning, etc.
- Mortality of control: 0 % (control), 0% (vehicle control)
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The substance deposited at the bottom of the tank.

Reported statistics and error estimates:

A Probit analysis was not possible with the available data. Therefore, a range was provided as LC50 value.

Any other information on results incl. tables

Sublethal observations / clinical signs:

ANALYTICAL RESULTS

The measured substance concentrations were > 80% of the nominal concentrations (Table 1). All results correspond to the degradation product. Degradation was complete after 2 h.

Table 1. Measured concentrations in test solutions.

Nominal	Mean measured concentration
[mg/L]	[mg/L]	%
10	9.4	94.0
22	22.3	101.4
50	50.2	100.4
100	95.9*	95.6

* mean of 0 and 48 h

 

BIOLOGICAL RESULTS

Due to the deep orange colour of the test solutions the observation of symptoms was limited in the 22 – 100 mg/L treatments. At the tested concentrations, altered swimming behavior and impaired breathing was observed. Some fish also showed alterations in behavior and physical aspect. Dead fish paled, had reddened gills and substance deposition on their body surface.

Table 2. Mortality after 96 h.

Concentration	Mortality
[mg/L]	Absolute	Relative [%]
0	0/30	0
0*	0/10	0
10	0/10	0
22	0/10	0
50	8/10	80
100	10/10	100

0* = vehicle control with 0.095 mL Tween 80/L