Fatty acids C8-18 (even numbered), mono- and diesters with sucrose

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:

06 - 24 May 1996

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:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

1992

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids C8-18 (even numbered), mono- and diesters with sucrose

EC Number:

947-862-2

Molecular formula:

not applicable, substance is UVCB

IUPAC Name:

Fatty acids C8-18 (even numbered), mono- and diesters with sucrose

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control, 22 and 50 mg/L
- Sampling method: In order to determine the substance concentration, water samples were taken from the middle of the test chamber by pipetting approx. 20 mL into a screw-neck glass bottle at study start, and after 48 and 96 h.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test substance was weighed into a beaker. Water for dilution was added, the mixture was homogenized using an Ultra-Turrax and poured into the chamber under stirring with a glass rod.
- Eluate: no
- Differential loading: yes
- Controls: yes, test medium control

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Source: Charles River Aquatics, Someren, Netherlands (date of hatching: 31 Jan 1996; Delivery date: 22 Apr 1996)
- Length at study initiation (length definition, mean, range and SD): 2.5 - 3.2 cm (mean: 2.9 cm ± 0.20 cm)

ACCLIMATION
- Acclimation period: 12 d
- Acclimation conditions (same as test or not): same as test
- Type and amount of food during acclimation: ad libitum; Tetra Min, Tetra Werke, Melle, Germany
- Feeding frequency during acclimation: twice daily

FEEDING DURING TEST : none

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Test temperature:

21.2 - 22.2 °C

pH:

start: 7.6 - 8.2
end: 8.0 - 8.1

Dissolved oxygen:

7.2 - 10.0 mg O2/L

Nominal and measured concentrations:

Nominal: control, 10, 22 and 50 mg/L (Concentrations were tested separately and not in parallel)
Measured: < LOQ, not measured, 19.13, 45.15 mg/L (mean measured)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass test vessels (10 L (30 x 22 x 24 cm) in a water bath). The temperature of the water was regulated by a thermostat.
- Aeration: no
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water, composition according to ISO/DIS 7346/1
- Culture medium different from test medium: same as test

OTHER TEST CONDITIONS
- Photoperiod: 12 h light/12 h darkness
- Light intensity: approx. 700 lux

EFFECT PARAMETERS MEASURED
- lnspection of the fish took place after 3, 6, 24, 48, 72 and 96 h and involved recording the lethality and visible changes in appearance and behavior. Dead fish were removed from the chambers. Fish were considered dead when there was a lack of opercular movement and no response to slight mechanical stimulus. The water parameters were measured and recorded before study start and after 0, 24, 48, 72 and 96 h.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: approx. 2.3

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

33 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: geometric mean of LC0 and LC100

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

13.2 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: re-calculated to 100% a.i.

Effect concentrations 3

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 4

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: The fish of the 50 mg/L group showed changes in appearance, behavior and swimming behavior. The 22 mg/L group showed no effect in comparison with the negative control while in the 10 mg/L group slight effects were observed only on the first day of exposure.
- Mortality of control: 0%
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Effect concentrations were clear and colorless. However, substance deposits on water and surface and bottom were recoded in some exposure vessels.

Reported statistics and error estimates:

Probit analysis could not be carried out because of the lethality rate. As the factor for the concentration steps was 2.2 (third root of 10), no further concentrations were tested. The geometric mean of the LC0 and LC100 is given.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Table 1: Mortality after 0, 48, 72 and 96 h

Concentration [mg/L]	24 h		48 h		72 h		96 h
	Absolute	%	Absolute	%	Absolute	%	Absolute	%
10	0/7	0	0/7	0	0/7	0	0/7	0
22	0/7	0	0/7	0	0/7	0	0/7	0
50	7/7	100	7/7	100	7/7	100	7/7	100

The measured concentrations were within the range of +/- 20% of nominal. Thus, the effect concentrations were based on the nominal concentrations.

Table 2: Validity criteria for OECD 203.

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	0%	yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	> 80%	yes
There must be evidence that the concentration of the substance being tested has been satisfactorily maintained, and preferably it should be at least 80 per cent of the nominal concentration throughout the test. If the deviation from the nominal concentration is greater than 20 per cent, results should be based on the measured concentration.	87-90%	yes

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.