Reaction products of stearic acid with 2-aminoethanol, maleic anhydride and sodium sulphite

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:

From December 13th, 2018 to March 08th, 2019

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The test item is a complex substance composed of various organic constituents having substantially different chemical structures. As the potential ecotoxicity of the test item cannot be assigned to a specific constituent, no appropriate analytical method for an accurate analysis could be developed and the biological endpoints were therefore expressed based on nominal test concentrations (loading rates).

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

The temperature exceeded the required range of 23±2°C for approximately 7 hours (max value: 25.3°C). Besides, the food was withheld from fish for three days prior start of the exposure (instead of one or two days).

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Remarks:

Analytical confirmation of the test concentrations were not performed due to technical reasons. As the test item is a complex substance composed of various organic constituents, no appropriate analytical method for an accurate analysis could be developed.

Vehicle:

no

Details on test solutions:

A stock solution at a nominal loading rate of 398.4 mg/L was prepared by adding 4185.0 mg test item to 10500 mL test medium. This stock solution was stirred for 22 h and 30 min at 700 rpm at ambient temperature in the dark. After 20 minutes without stirring, particles were observed. Thereafter, the stock solution was filtered, using a glass fibre filter with an average retention capacity of 0.4 pm (MN 85/220, diameter: 110 mm). The first 0.4 L of the filtrate were discarded, visual inspection of the filtrate showed no undissolved test item particles. The filtrate (stock solution, S2) was used to prepare the desired test item concentration and was used immediately after preparation.

To prepare the limit test concentration the stock solution S2 (filtrate) was directly provided in the test vessel. Test medium without test item was used for the control.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Source: ECT (stock culture).
- Spawning date: September 27th, 2018.
- Length at study initiation: range: 2.04-2.49 cm , mean: 2.21 cm (assessed from a representative sub-sample consisting of 10 fish).
- Weight at study initiation: range: 0.0634-0.0985 g/fish, mean: 0.0826 g/fish (assessed from a representative sub-sample consisting of 10 fish).

ACCLIMATION
- Type and amount of food during acclimation: TetraMin® and artemia nauplien.
- Feeding frequency during acclimation: daily ration fed in 3 portions on workdays, but not within three days before start of exposure.
- Health during acclimation: no mortalities observed during the 7-day period before the start of the test.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

169.67 mg CaCO3/L

Test temperature:

21.1-25.3°C

pH:

7.4-7.6

Dissolved oxygen:

7.2-8.1 mg/L (81-92% of air saturation value)

Conductivity:

803 µS/cm2

Nominal and measured concentrations:

Nominal concentration (loading rate): 398.4 mg test item/L, equivalent to 100 mg active ingredients/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: not specified.
- Type (open / closed ): not specified.
- Fill volume: 10 liters.
- Aeration: permanent.
- No. of organisms per vessel: 7.
- No. of vessels per concentration (replicates): 1.
- No. of vessels per control (replicates): 1.
- Biomass loading rate: 0.058 g/L.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water accoridng to OECD Guideline 203 (modified as described below).
The reconstituted water consisted of deionised water and salts in the following final concentrations:
A) CaCl2 x 2 H20 294.0 mg/L
B) MgSO4 x 7 H20 123.0 mg/L
C) NaHCO3 64.8 mg/L
D) KCI 5.75 mg/L
Prior to use, the reconstituted water was modified, i.e. diluted with deionised water (1:1; v/v), and supplemented with 1% of artificial seawater (Tropic Marin, Dr. Biener GmbH, Wartenburg, Germany; Atkinson MJ and C Bingman (1998)). The required amount of modified reconstituted water was prepared within one month before use. During storage, the water was aerated. After aeration of several hours, the physico-chemical characteristics were determined according to the requirements of the test guideline.

OTHER TEST CONDITIONS
- Adjustment of pH: none.
- Feeding: none.
- Photoperiod: 14h light / 10h dark.
- Light intensity: 100-1000 lx.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- All fish in the test were observed for mortality 3, 6, 24, 48, 72 and 96 hours after start of exposure.
- All fish in the test were observed daily for any effects other than lethality: effects on the appearance, size and behaviour of the fish that make them clearly distinguishable from the control animals, e.g. different swimming behaviour, changes in appearance of the fish were recorded.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 398.4 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:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

There was no mortality, neither in the control nor the test item treatment (limit concentration at 100% filtrate of a saturated solution at a loading rate of 398.4 mg test item/L equivalent to 100 mg active ingredients/L) during the 96-hour exposure period. There were no behavioural differences in fish in the limit concentration compared with the control fish.

Sublethal observations / clinical signs:

Up to and including the limit test concentration (100% filtrate of a saturated solution at a loading rate of 398.4 mg test item/L equivalent to 100 mg active ingredients/L), no mortalities were observed during the 96-hour exposure period. Therefore, it can be concluded that the LC50 (96 h) to fish (Danio rerio) based on nominal concentrations is higher than 398.4 mg test item/L, equivalent to 100 mg active ingredients/L.

Validity criteria fulfilled:

yes

Remarks:

All validity criteria were fulfilled as required by the study plan and the guideline (mortality in the control: ≤10%; dissolved oxygen concentration in control and test vessels: ≥60% of air saturation value).

Conclusions:

Up to and including the limit test concentration (100% filtrate of a saturated solution at a loading rate of 398.4 mg test item/L equivalent to 100 mg active ingredients/L), no mortalities were observed during the 96-hour exposure period.

Executive summary:

The acute toxicity to the zebra fish Danio rerio of the REACH substance "Stearic acid, reaction product with 2-aminoethanol, maleic anhydride and sodium sulfite" (UVCB) was investigated in a GLP-compliant study performed according to OECD Guideline 203. Up to and including the limit test concentration (100% filtrate of a saturated solution at a loading rate of 398.4 mg test item/L equivalent to 100 mg active ingredients/L), no mortalities were observed during the 96-hour exposure period. Therefore, it can be concluded that the LC50 (96 h) to fish based on nominal concentrations is higher than 398.4 mg test item/L, equivalent to 100 mg active ingredients/L.

Description of key information

The acute toxicity to the zebra fish Danio rerio of the REACH substance "Stearic acid, reaction product with 2-aminoethanol, maleic anhydride and sodium sulfite" (UVCB) was investigated in a GLP-compliant study performed according to OECD Guideline 203. Up to and including the limit test concentration (100% filtrate of a saturated solution at a loading rate of 398.4 mg test item/L equivalent to 100 mg active ingredients/L), no mortalities were observed during the 96-hour exposure period. Therefore, it can be concluded that the LC50 (96 h) to fish based on nominal concentrations is higher than 398.4 mg test item/L, equivalent to 100 mg active ingredients/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

One reliable experimental study is available and is selected as key study (ECT, 2019). It was performed according to OECD Guideline 203 and under GLP principles.