Reaction mass of (9-acetoxy-3,8,10-triethyl-7,8,10-trimethyl-1,5-dioxa-9-azaspiro[5.5]undec-3-yl)methyl palmitate and (9-acetoxy-3,8,10-triethyl-7,8,10-trimethyl-1,5-dioxa-9-azaspiro[5.5]undec-3-yl)methyl stearate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

The degradation products of the test item were analytically determined. The test item could not be analyzed due to the low solubility and the fast
degradafion of. Thus, the resulting degradation products will be more relevant for any toxic effects on the test organisms under natural conditions.

For the analysis of the actual test item concentrafions the following samples were taken:
just before test start:
- duplicate samples from the test medium
- duplicate samples from the control

after 48 and 96 hours:
- duplicate samples from the test medium
(stability samples) - duplicate samples from the control

All samples were taken from the approximate center of the aquaria without mixing the test media.

The concentration of the degradation products was analyzed in the duplicate test medium samples from all sampling fimes (0, 48 and 96 hours).
From the control samples only one of the duplicate samples was analyzed from the corresponding sampling fimes. The samples were analyzed
immediately after sampling without prior storage.

Vehicle:

no

Details on test solutions:

According to the results of a pre-experiment (without GLP) the test item was not soluble at a concentration of 100 mg/L in test water and no
homogeneous dispersion could be prepared. A filtrate of a supersaturated dispersion of the test item was prepared. The undiluted filtrate
was tested as the only test concentration. Additionally, a control was tested in parallel (test water without test item). A supersaturated dispersion with a nominal concentration of 100 mg/L (= loading rate) was prepared by weighing 700 mg of the test item into 7 liters test water. No auxiliary solvent
or emulsifier was used. The test item was mixed into the test water as homogeneously as possible by ultrasonic treatment for 15 minutes and by
intense stirring for 96 hours at room temperature in the dark to dissolve a maximum concentrafion of the the test item in the dispersion.
The long stirring period of 96 hours was chosen because the concentrafion of dissolved degradation product continuously increased over a stirring
time of about 96 hours (results of a pre-test; without GLP).

The supersaturated stock dispersion of the test item was filtered through a membrane
filter (Schleicher & Schuell, Type NC45, pore size 0.45 microm) after the 96 hours stirring period just before the preparafion of the test medium. The undiluted filtrate of the supersaturated dispersion with the maximum concentrafion of dissolved degradation product was used as the only test
medium.

The test medium was prepared just before introducfion of the fish (= start of the test).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Source: Zoohaus Schaub, CH-4410 Liestal, Switzerland
- Age at study initiation (mean and range, SD): 3.4 +- 0.1 cm
- Weight at study initiation (mean and range, SD): 0.41 +- 0.03 g
- Feeding during test: none


ACCLIMATION
- Acclimation period: 1 week
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: commercial fish diet (TETRA MIN Hauptfutter)
- Health during acclimation (any mortality observed): no mortality observed

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

2.5 mmol/L (=250 mg/L) as CaCO3

Test temperature:

22 deg. C

pH:

7.9

Dissolved oxygen:

> 8.3 mg/L

Nominal and measured concentrations:

loading rate of 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 5 L glass aquaria
- Aeration: slight aearation during test period
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: < 1 g fish wet wgt per liter test medium

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water
- Ca/mg ratio: 4:1
CaCl2 X 2H2O 2.0 mmol/L (= 294 mg/L)
MgSO4 X 7H2O 0.5 mmol/L (=123 mg/L)
NaHCO3 0.75 mmol/L (= 65 mg/L)
KCI 0.075 mmol/L (= 5.8 mg/L)
Water Hardness 2.5 mmol/L (= 250 mg/L) as CaCO2
Alkalinity 0.8 mmol/L
Na : K = 10 : 1 (based on molarity)





OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod:
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:
- Results used to determine the conditions for the definitive study:

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

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:

- Behavioural abnormalities: none
- Mortality of control: none
- Other adverse effects control: none

Sublethal observations / clinical signs:

In conclusion, the test substance and its degradation products had no toxic effect on

the fish up to their solubility limit in test water under the present test condifions.

Description of key information

With high probability acutely not harmful to aquatic organisms. No toxic effects were observed within the range of solubility.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

>= 100 mg/L

Additional information

According to regulation (EC) No 1907/2006, Annex XI, paragraph 1.5., substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group, or "category" of substances. Environmental effects or environmental fate may be predicted from data for reference substance(s) within the group by interpolation to other substances in the group (read-across approach). The substance consists of two main components, a component with a C18 chain (60%) and C16 chain (30%) component. The structure of both substances is the same except the chain length.

The substance with C18 is already notified under EC 445-990-9. It is expected that the structural difference between the octadecanoate and the hexadecanoate does not affect the physico-chemical properties, the toxicological and ecotoxicologocal profile as well as the environmental fate. Furthermore, it can be assumed that the degradation products of EC 445-990-9 are very similar to the degradation products of the mixture containing the octadecanoate and the hexadecanoate. Therefore read-across to EC 445-990-9 is scientifically justified for physico-chemical properties, the toxicological and ecotoxicologocal profile as well as environmental fate.