Reaction products resulting from esterification of sucrose with saturated C16-18 (even numbered) fatty acids

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990-1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No test substance analytics

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

adopted 04.04.1984

Deviations:

yes

Remarks:

species (Leuciscus idus) different fron those recommended in OECD guideline 203

Principles of method if other than guideline:

It was used a fish species not recommended in OECD Guideline 203, but in comparable EU-method.

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- The test substance is slightly soluble in water. To set the appropriate test concentrations, the necessary substance amounts were weight in small plastic plates and put into aquariums filled with 10 L water. The substance is homogenized by stirring.
- The dilution water is mixture of Cologne drinking water and deionized water (1:1).
- The dilution water was areated up to oxygen saturation before use.

Test organisms (species):

Leuciscus idus

Details on test organisms:

TEST ORGANISM
- Scientific name: Leuciscus idus
- Common name: Golden Orfe
- Source: Fischzucht (fish farm) Eggers, Hohenwested, Germany
- Age at study initiation (mean and range, SD): ca. 4 weeks

ACCLIMATION
- Acclimation period: minimum 7 days
- Acclimation conditions: in tap water, temp.: 20 +/- 0.5 °C, oxygen content: min. 80 % of saturation value, photoperiod: 12 h/day, no of organisms per vessel: < 3.7
- Type and amount of food: Pitti Floxi-Flocken Alleinfutter für Zierfische
- Feeding frequency: twice per week
- Health during acclimation (any mortality observed): no data

Test type:

static

Water media type:

freshwater

Total exposure duration:

96 h

Hardness:

15 ± 3 ° dH

Test temperature:

20 ± 0.5 °C

pH:

7.8 - 8.1

Dissolved oxygen:

6.1 - 10.3 mg/L

Salinity:

-

Nominal and measured concentrations:

Nominal 2000, 3000, 4000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: glass (length: 28.5 cm x width: 21 cm x height: 24 cm)
- Aeration: slight aeration
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: mixture out of tap water (Köln, Germany) and deionized water
- Ca/Mg ratio: 4:1
- Intervals of water quality measurement: 24, 48, 72, 96 h

OTHER TEST CONDITIONS
- Adjustment of pH: no
During the test the fishes were not fed.

EFFECT PARAMETERS MEASURED: The mortality was observed after an exposure time of 96h.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

2 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

4 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality at 96 h:
0% with 2000 mg/l
20% with 3000 mg/l
100% with 4000 mg/l
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no

Sublethal observations / clinical signs:

During the test the following mortality rates were observed:

Exposition time (h)	Mortality abs./rel %	Control	Concentration (mg/L)
			2000	3000	4000
24	abs	0/10	0/10	0/10	0/10
	rel %	0	0	0	0
48	abs	0/10	0/10	0/10	0/10	10/10
	rel %	0	0	0	0	100
72	abs	0/10	0/10	1/10	5/10	---
	rel %	0	0	10	50
96	abs	0/10	0/10	2/10	10/10	---
	rel %	0	0	20	100

Oxygen Content

 

Concentration (mg/L)	Exposition time (h)
	24	48	72	96
2000	9,2	9,0	7,3	10,3
3000	9,5	7,8	8,1	6,1
4000	9,9	9,0	7,3	10,3

Validity criteria fulfilled:

yes

Conclusions:

In a 96-h acute toxicity study according to OECD TG 203, Leuciscus idus were exposed to Fatty acids, C16 and C18, esters with sucrose, glycerol and ethylene glycol at nominal concentrations of 0 (control) 2000, 3000 and 4000 mg/L under static conditions. The test substance was directly transferred into the dilution water. No analytical control was performed.
The 96-h LC50 is 3100 mg/L (arithmetical determination), the 96-h LC0 is 2000 mg/L, the 96-h LC100 is 4000 mg/L. The reliability was assessed 2 because no analytical control was performed.

Executive summary:

In a 96-h acute toxicity study, Leuciscus idus (Golden Orfe) were exposed to Fatty acids, C16 and C18, esters with sucrose, glycerol and ethylene glycol at nominal concentrations of 0 (control), 2000, 3000 and 4000 mg/L under static conditions. The 96-h LC50 was 3100 mg/L (arithmetical determination). The LC0 and LC100 values, based on mortality, were 2000 and 4000 mg/L, respectively.

This toxicity study is classified as reliable with restrictions as any analytical monitoring was performed.

The following validity criteria are fulfilled: the mortality in the controls did not exceed 10% and constant test conditions were maintained throughout the test.

 

Reference 2

Endpoint:

fish embryo acute toxicity (FET)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 236 (Fish embryo acute toxicity (FET) test)

Version / remarks:

2013

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 012661240
- Expiration date of the lot/batch: 01 Sep 2018
- Purity: 84.1% active matter

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: One water accommodated fraction (WAF) was prepared 24 ± 1 hour prior to the start of the exposure. The test item is sparingly soluble and a complex mixture (UVCB). The limit concentration was therefore prepared as loading. An appropriate amount of the test item was grounded, weighed and transferred into a flask with an appropriate amount of dilution water to obtain a concentration of 120 mg/L. The solution was stirred for 24 hours at approx. 1100 rpm at 30 °C. After stirring, undissolved particles were removed by membrane filtration (membrane filter 0.45 µm, RC, Macherey-Nagel). The filter was saturated in order to avoid adsorption during the filtration. The first 25 mL of the filtrate were discarded. The filtration was interrupted for 15 minutes to for allow adsorption and saturation of the filter material with dissolved test item. Thereafter, the filtration was continued. The next 25 mL were discarded. The following water soluble fraction (WSF) was used in the test. During filtration, the filter was always kept covered. The test solution was aerated for approx. 30 minutes before use. This preparation procedure was repeated prior to the start of every interval. The test vessels were saturated with test solutions of the control and limit loading for at least 24 hours before the start of the exposure.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: All fish used for brood fish were gained at the test facility from a single brood stock (supplier: Umweltbundesamt, Schichauweg 58, 12307 Berlin, Germany)
- Method of breeding and maintenance of the brood fish: A breeding stock of unexposed, mature zebrafish with an age of 9 months was used for egg production. Fish were free of macroscopically discernable symptoms of infection and disease. Spawners were maintained in aquaria with a loading capacity of a minimum of 1 L water per fish: temperature: 25 °C; dissolved oxygen concentration > 60 % of air saturation value; pH-value: 6.5 – 8.5; photoperiod: 16 h light / 8 h dark cycle; Diffuse light (7 – 750 Lux on water surface); Food: Artemia salina nauplii, 48 hours old, ad libitum; Daphnia magna, juvenile and adult daphnids, ad libitum; dry food sera vipan SERA, ad libitum; no disease treatments were administered.
Adult zebrafish were kept in separate aquaria. About 15 minutes before the start of artificial dawning, spawn traps (rectangular dishes (26 cm x 14 cm x 6 cm), covered with a stainless steel mesh and provided with artificial plants) were introduced into the aquaria. After the end of dawning (approximately 0.5 h), the glass dishes were gently removed. About 150 eggs were taken, washed gently with dilution water and immediately distributed to the test concentrations and the control (40 eggs for the control, the reference item and for the test item loading).
- Fertilization check: Approximately 2 h post fertilization, eggs were checked for fertilization. Every embryo was checked under a stereo microscope (40-fold magnification) for its blastomer phase. Eggs with only a 2 cell blastomer were regarded as not fertilised. These eggs as well as coagulated eggs were discarded.
- Fertilization rate: The overall fertilization rate of all collected eggs was ≥ 95 %.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

60-66 mg/L as CaCO3

Test temperature:

26 ± 1 °C

pH:

7.23 - 7.81

Dissolved oxygen:

Not less than 80% of air saturation value

Conductivity:

169 µS/cm

Nominal and measured concentrations:

Nominal: 120 mg/L as loading rate

Details on test conditions:

TEST SYSTEM
- Test vessel: 24 well micro-titre plates containing 2 mL test solution
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 20
- No. of vessels per control (replicates): 20

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Maintenance water was used. The water was filtered on activated charcoal and aerated for at least 30 minutes.:
- Intervals of water quality measurement: The water quality (pH-value, oxygen saturation, temperature) was measured daily in a surrogate chamber per limit loading and control from the freshly prepared and corresponding 24 hours aged test solutions, respectively. Total water hardness was measured before the beginning and after the end of exposure from dechlorinated water. The water temperature was measured continuously (once per hour) with a data logger in a separate glass beaker.

OTHER TEST CONDITIONS
- Photoperiod: The test was incubated in the dark, except for observation

EFFECT PARAMETERS MEASURED: Coagulation of embryos, absence of somite formation, non-detachment of the tail and lack of heart-beat are lethal endpoints and were recorded after 24, 48, 72 and 96 h.

Reference substance (positive control):

yes

Remarks:

3,4-Dichlororaniline

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(WAF)

Basis for effect:

coagulation of the embryo

Details on results:

The test substance did not cause any effects on embryonic stages of zebrafish.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: 96h-LC50 = 3.34 mg/L

Validity criteria fulfilled:

yes

Conclusions:

The test substance is not acutely harmful to embryonic stages of zebrafish.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No test substance analytics

Justification for type of information:

Both target and source substance are Sucrose ester with long-chain fatty acids. In addition, the source substance contains glycerol and ethylene glycol moieties. Since these substances, as the long-chain fatty acids, are not harmful to aquatic organisms, this structural difference is not considered to alter the ecotoxicity of the source substance in comparison to the target substance.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Conclusions:

In a 96-h acute toxicity study according to OECD TG 203, Leuciscus idus were exposed to Fatty acids, C16 and C18, esters with sucrose, glycerol and ethylene glycol at nominal concentrations of 0 (control) 2000, 3000 and 4000 mg/L under static conditions. The test substance was directly transferred into the dilution water. No analytical control was performed.
The 96-h LC50 is 3100 mg/L (arithmetical determination), the 96-h LC0 is 2000 mg/L, the 96-h LC100 is 4000 mg/L. The reliability was assessed 2 because no analytical control was performed.

Executive summary:

In a 96-h acute toxicity study, Leuciscus idus (Golden Orfe) were exposed to Fatty acids, C16 and C18, esters with sucrose, glycerol and ethylene glycol at nominal concentrations of 0 (control), 2000, 3000 and 4000 mg/L under static conditions. The 96-h LC50 was 3100 mg/L (arithmetical determination). The LC0 and LC100 values, based on mortality, were 2000 and 4000 mg/L, respectively.

This toxicity study is classified as reliable with restrictions as any analytical monitoring was performed.

The following validity criteria are fulfilled: the mortality in the controls did not exceed 10% and constant test conditions were maintained throughout the test.

 

Description of key information

LL50 (96h) > 120 mg/L (nominal) for Danio rerio (OECD 236)

Additional information

One study investigating the short-term toxicity of Reaction products resulting from esterification of sucrose with saturated C16-18 (even numbered) fatty acids to fish is available. The study was performed according to GLP and OECD guideline 236 using embryonic stages of zebrafish as test organisms (Herrmann 2018). A nominal test concentration of 120 mg/L was tested as loading rate in a limit test. No coagulation of embryos, absence of somite formation, non-detachment of the tail or lack of heart-beat were recorded throughout the exposure time in control and treatment. Thus, the 96h-LL50 is determined to be > 120 mg/L based on nominal concentrations.

The conclusion that the test substance is not harmful to fish is confirmed by an additional study performed with the structurally related source substance Fatty acids, C16 and C18, esters with sucrose, glycerol and ethylene glycol was conducted. Both substances are Sucrose ester with long-chain fatty acids. In addition, the source substance contains glycerol and ethylene glycol moieties. Since these substances, as the long-chain fatty acids, are not harmful to aquatic organisms, this structural difference is not considered to alter the ecotoxicity of the source substance in comparison to the target substance.

The study with the source substance was performed according to OECD guideline 203 using Leuciscus ideus as test organism (Evonik 1991). Nominal concentrations between 2000 and 4000 mg/L were tested. No analytical monitoring was performed. The 96h-LC50 was determined to be 3100 mg/L based on nominal concentrations.