reaction product of: saturated, monounsaturated and multiple unsaturated long-chained partly estrified alcohols of vegetable origin (Brassica napus L., Brassica rapa L., Helianthus annuus L., Glycine hispida, Gossypium hirsutum L., Cocos nucifera L., Elaeis guineensis) with O,O-diisobutyldithiophosphate and 2-ethylhexylamine and hydrogen peroxide

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:

Jan 27/1998 to Jan 31/1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted in compliance with agreed test protocols following guidance.

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

428-630-5

EC Name:

-

Molecular formula:

Not applicable

IUPAC Name:

reaction product of Z-9-octadecen-1-ol and O,O-diisobutyl hydrogen dithiophosphate

Test material form:

other: liquid

Details on test material:

- Substance type: orange-yellow, clear liquid
- Physical state: orange-yellow, clear liquid
- Lot/batch No.: 12447
- Storage condition of test material: in a well closed container and protected from light at room temperature

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
None.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The amount of the test substance dissolved in water was checked by DOC analysis (Dissolved Organic Carbon) made at the beginning of the test, after 48 hours and after 96 hours. Samples taken fkom each test vessel were centrifkged at 3,500 rpm for 10 minutes in order to separate the substance unsolved. The concentration of DOC in the clear aqueous phase was determined.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The nominal concentration of the test group was 100 mg/l. 400 mg test substance and 4L test water were stirred for 24 hours. The mixture of water and test substance were used for the test

- Eluate: reconstituted water (Potassium chloride 5.75 mg/l; Calcium chloride Dihydrate 294.00 mg/l; Magnesium sulphate Heptahydrate 123.25 mg/l; and Sodium bicarbonate 64.75mg/l).
- Differential loading: not applicable.
- Controls: test medium.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no information is available.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: Labor- und Zierfischzuchtbetrieb Gunter Rosenhainer
- Age at study initiation (mean and range, SD): no data
- Length at study termination (length definition, mean, range and SD): 2.6 - 3.5 cm
- Weight at study termination (mean and range, SD): no data

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions (same as test or not): yes
- Type and amount of food: fiozen larvae of mosquitoes
- Feeding frequency: daily and discontinued 24 hours prior to the test period

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

none

Test conditions

Hardness:

none

Test temperature:

2 1.8-22.4 deg C

pH:

7.4-7.9

Dissolved oxygen:

8.0 - 8.5 mg O2/L.

Salinity:

not applicable.

Nominal and measured concentrations:

nominal concentrations: 0 and 100 mg/L

Details on test conditions:

TEST SYSTEM

- Type (delete if not applicable): static
- Aeration: at least 8.6 mg/l; the vessels were aerated
- No. of organisms: 1 fish /Liter
- Biomass loading rate: 0.39 g bw/L


OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: 15 hours light, 9 hours dark
- Light intensity: no data

TEST CONCENTRATIONS
- Range finding study: none.
- Test concentrations: 0 and 100 mg/L

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

There were no mortalities in fish exposed to a 100 mg/l test substance for a period of 96 hours.

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:

Table 1: Cumulative mortality data in the definitive study.

Conc. (mg/L)	Cumulative mortality						% mortality
	0 h		24 h	48 h	72 h	96 h	96 h
Control	0		0	0	0	0	0
100	0		0	0	0	0	0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 96-Hour LC50 zebrafish based on nominal concentration was greater than 100 mg/L.

Executive summary:

Introduction:A study was performed to assess the acute toxicity of the test material to Zebra-fish (Brachydanio rerio). The method followed that described in the guideline 92/69/EEC, part C. 1.

Methods:Zebra fish was exposed to the test material, at a single nominal concentration of 100 mg/L for a period of 96 hours under static test conditions. The number of mortalities and any sub-lethal effects of exposure in test and control vessel were determined 6 hours after the start of exposure and then daily throughout the study until termination after 96 hours.

The dissolved oxygen concentration, pH values and temperature were checked daily.

Results:

The concentration of the substance in test water was measured by determination of the Dissolved Organic Carbon (DOC). At the start of the test the concentration was 3.8 mg/l and at the end of the exposure period the value was 4.5 mg/l.

No mortalities occurred during the test in the control and the test group. The fish showed no visible abnormalities. The 96-Hour LC50 based on nominal concentration was greater than 100 mg/L. It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L. The substance is not acute toxic to fish.