Soybean oil, epoxidized, Me ester, reaction products with propylene glycol

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:

12 November 2012 - 26 November 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

17th July 1992

Qualifier:

according to guideline

Guideline:

other: OECD Guidance Document 126: Short Guidance on the Threshold Approach for Acute Fish Toxicity

Version / remarks:

Adopted 31 May 2010

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Test substance: Sovermol 1055
Chemical name: Fatty acids, tallow, Me esters, epoxidized
BMG Sample no: M1204-03199-01
BASF project no: 17F0029/12X046
BASF substance no: 12/0029-1
Batch no: CE80580015
CAS no: 158318-67-3
Physical form: Liquid
Water solubility: insoluble
Purity: >90%, treated as 100% pure (the exact value is currently being analyzed, BASF study code 12L00069)
Test substance storage:At room temperature, protected from light
Expiry date:27 February 2013

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100 mg/L
- Sampling method: 0, 24, 48, 72 and 96 hour

Vehicle:

no

Details on test solutions:

Since Fatty acids, C16-18 and C18-unsatd., Me esters, epoxidized is a multi-component substance that is only partially soluble, the water accommodated fraction (WAF) approach, based on OECD guidance document N0.23 (2000), was used. The loading of the test material was prepared by adding the respective amount of an acetonic stock solution to an empty glass vessel. After complete evaporation of the solvent, the natural water was added, moderately stirred for 72 h, followed by filtration (MILLIPORE AP15 glass fiber filter). The resulting water soluble fraction (WSF) was used in the test.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: not specified
- Age at study initiation: Not specified
- Length at study initiation: total length of 12-17 mm
- Weight at study initiation: 31 - 76 mg
- Culturing: Population of young fish are held in 40 l glass aquaria. The bottom of the aquaria is covered with gravel (≤1.5 cm).

ACCLIMATION
- Acclimation period: Fish were obtained from a reliable commercial supplier. Fish were gradually acclimated to the test conditions, and were held for at least an additional 12 days prior to testing in water of similar temperature (23°C ± 2.0 °C) and quality used in the test.
- Acclimation conditions (same as test or not): yes
- Type and amount of food during acclimation: The fish were fed commercially prepared diet Sera micropan and Sera vipan (Sera apanristic GmbH, D- 52525 Heinsberg, Germany) until 24 hours prior to the exposure.
- Health during acclimation (any mortality observed): During the present acclimatization period, no mortality was observed.

OTHER
- Handling: The exposure began, when all the fish were randomly placed in the test vessels. The transfer was performed in an impartial manner to eliminate bias. Fine-mesh dip net was used to transfer fish, taking care to minimise possible stress due to handling. Fish which were damaged or dropped during the transfer were not used.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

Test start (t=0h): 22.5 °C
Test end (t=96h): 23.0 °C

pH:

Test start (t=0h): 8.0 - 8.2
Test end (t=96h): 8.6 - 8.7

The pH in the blank control was in the range of 8.0-8.6. Although the pH was not in the recommended range of 6.0-8.5, the deviation had no adverse effect as no mortality was observed in the blank control.

Dissolved oxygen:

Test start (t=0h): 6.2 - 6.3 mg/L
Test end (t=96h): 7.0 mg/L

The dissolved oxygen concentration was ≥6.2 mg O2/l corresponding to an air saturation of ≥75% at 24°C; a concentration of 8.25 mg/L corresponds to 100% saturation at 24°C (OECD Guideline 203: The dissolved oxygen concentration must have been at least 60% of the air saturation value throughout the test).

Nominal and measured concentrations:

Nominal loading rate: Blank control and 100 mg/L

Measured concentrations: HPLC analysis of the test substance concentration was conducted at the beginning and after 24, 48, 72 and 96 h of exposure. The measured concentrations were 4.39, 4.42, 4.34, 4.38 and 4.44 mg/l, respectively. The geometric mean concentration over a period of 0-96 h (GM0-96 h) was 4.39 mg/l (100% of initial dissolved concentration).

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria of 2 l (14.5 x 14.0 x 9.5 cm) filled with 1.0 liter of water.
- Aeration: Continuously aerated tap water, the total dissolved oxygen concentration was kept >60% of the saturation value with aeration via Pasteur pipettes.
- Number of fish: 7 fish per test concentration and control

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Continuously aerated tap water of the town of Schlieren

Parameter a) Unit Minimum Maximum
pH 7.0 b) 7.5 b)
Conductivity (25°C) µS/cm2 400 b) 700 b)
Total Hardness °f H 16.1 20.9
mmol/l 1.61 2.09
mg/l 161 209
CO3 – Hardness °f H 14.6 18.8
mg/l 146 188
Rest – Hardness °f H 1.5 2.1
mg/l 15 21
Acid consumption mmol/l 2.92 3.76
E. Coli per 100 ml 0 0
Colony counts: (20°C) per ml 0 3
(30°C) 3 14
Ammonium mg/l <0.01 <0.01
Nitrite mg/l <0.002 <0.002
Nitrate mg/l 4.0 6.0
Chloride mg/l 7.0 11.5
Phosphate as PO4-P mg/l 0.019 0.037
Sulfate mg/l 15.5 18.5

a) Analysis by Kantonales Labor Zürich, CH-8032 Zurich, 8 September 2011
b) Bimonthly analysis by BMG Engineering Ltd, starting date: September 2012
The parameters analyzed by the Kantonales Labor Zürich are based on the values obtained from three groundwater wells (aquifer of the river Limmat). These three wells represent the major fraction (90%) of the drinking water of the town of Schlieren. The remaining 10% derive from spring water. The water quality fulfills all legal requirements. Therefore, no chlorination is needed and the water can be used in the tests without any treatment.

- Intervals of water quality measurement: Measurements for oxygen, pH and temperature were made prior to the addition of the fish to the water soluble fraction (WSF) and after 24, 48, 72 and 96 h of exposure

OTHER TEST CONDITIONS
- Adjustment of pH: not adjusted
- Photoperiod: 16 h photoperiod a day, supplied by overhead white fluorescent tubes, no transition period.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Fish were considered to be dead if they showed no reaction after touching the caudal fin and no visible movement (e.g. gill movements). The median lethal concentration (LC50 value) is the concentration killing 50% of the fish after a certain period of exposure.

TEST CONCENTRATIONS
This study was performed as a limit test at the loading rate of 100 mg/l. The selected limit loading rate was based on the results of preceding non-GLP-tests with Daphnia (in the course of study no. A12-00820) and algae (in the course of study no. A12- 00819) yielding EC50 values of >100 mg/l loading rates.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Based on the binominal theory, the absence of mortality provides at least 99% confidence that the LC50 is greater than the applied loading rate of 100 mg/l

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 4.39 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Based on the binominal theory, the absence of mortality provides at least 99% confidence that the LC50 is greater than 4.39 mg/l geometric mean concentration (GM0-96 h) of Fatty acids, C16-18 and C18-unsatd., Me esters, epoxidized.

Duration:

96 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 4.39 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Mortality and other symptoms:
At the loading rate of 100 mg/l Fatty acids, C16-18 and C18-unsatd., Me esters, epoxidized, none of a total of seven fish died after 96 h of exposure. No mortality was observed in the blank control.
Toxic effects other than mortality, such as loss of coordination, hypo or hyperactivity and swimming on the back, were not observed.

Reported statistics and error estimates:

No statistical analysis was performed. Instead, the effect concentration (EC) values were estimated to be lower or higher than the tested loading rate based on the observed mortality.

Estimation of LC50: The absence of mortality provides at least 99% confidence that the LC50 is greater than the concentration used in the limit test, based on the OECD Guidance Document 126: Short Guidance on the Threshold Approach for Acute Fish Toxicity, adopted 31 May, 2010.

Determination of no observed effect concentration The no observed effect concentration (NOEC) is the highest test concentration for which the test item is observed to have no biologically significant effects when compared to the controls. Mortality rates of =<10% (1 fish) are regarded as not biologically significant.

Validity criteria fulfilled:

yes

Remarks:

Mortality in the control did not exceed 10%, dissolved oxygen concentration must have been at least 60% of the air saturation value throughout the test

Conclusions:

The 96-h LC50 value was determined to be above 100 mg/L (> 4.39 mg/l geometric mean concentration) in Danio rerio (Zebrafish) when exposed to Fatty acids, C16-18 and C18-unsatd., Me esters, epoxidized.

Executive summary:

The acute lethal toxicity of Fatty acids, C16-18 and C18-unsatd., Me esters, epoxidized to Danio rerio (Zebrafish) was investigated in a guideline OECD TG203 study under GLP and static exposure conditions over a period of 96 h. The singly applied loading rate was 100 mg/l. HPLC analysis of the test substance concentration was conducted in the definitive test at the beginning and after 24, 48, 72 and 96 h of exposure. The measured concentrations were 4.39, 4.42, 4.34, 4.38 and 4.44 mg/l, respectively. The geometric mean concentration over a period of 0-96 h (GM0-96 h) was 4.39 mg/l (100% of initial dissolved concentration). At the loading rate of 100 mg/l, none of a total of seven fish died after 96 h of exposure. No mortality was observed in the blank control. Toxic effects other than mortality, e.g. loss of coordination, hypo- or hyperactivity and swimming on the back, were not observed. The no-observed-effect concentration (NOEC) was >=100 mg/l loading rate and >=4.4 mg/l geometric mean concentration. Based on the binominal theory, the absence of mortality provides at least 99% confidence that the LL50 is greater than the applied loading rate of 100 mg/l and > 4.4 mg/l when based on geometric mean concentration.

Reference 2

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:

other: The result derived from read across is sufficiently reliable because all Annex XI criteria are met.

Justification for type of information:

A read-across approach appears appropriate to predict the endpoint “Fish acute toxicity” for the substance Soybean oil, epoxidized, methyl ester, reaction products with propylene glycol because:

 A fish 96-hour toxicity study, according to OECD test guideline 203, is available for the source substance: Fatty acids, C16-C18 and C18-unst, Me esters, epoxidized [EC 605-143-8, CAS 158318-67-3]
 Both the source and target substances are part of a chemical group consisting of Epoxidized Oils and Derivatives (EOD). The composition of the source chemical: Fatty acids, C16-C18 and C18-unst, Me esters, epoxidized, closely resembles the target substance.
 The epoxide value for the source substance is higher than for the target substance indicating that this substance is potentially more reactive and thus an acceptable (worst-case) candidate regarding the prediction of possible ecotoxicological effects.
 This prediction intends to provide acute fish toxicity information to assess the classification for aquatic environmental hazards according to the CLP Regulation EC/1272/2008.
 By means of this read-across approach, vertebrate animal testing is avoided.

This report follows the RAAF method and so presents:
1) The hypothesis: analogue read-across approach, based on the similarity of the substances and the absence of significant difference for these types of structures regarding the considered endpoint.
2) The scientific justifications (“Assessment Elements”) and their evaluation (“Assessment Options”); which demonstrate the confidence that can be put in this prediction.
3) The conclusions, usable for classification assessment or risk assessment, which are summarised.

Reason / purpose for cross-reference:

read-across source

Total exposure duration:

96 h

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Based on the binominal theory, the absence of mortality provides at least 99% confidence that the LC50 is greater than the applied loading rate of 100 mg/l

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 4.39 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Based on the binominal theory, the absence of mortality provides at least 99% confidence that the LC50 is greater than 4.39 mg/l geometric mean concentration (GM0-96 h) of Sovermol 1055.

Duration:

96 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 4.39 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Mortality and other symptoms:
At the loading rate of 100 mg/l Sovermol 1055, none of a total of seven fish died after 96 h of exposure. No mortality was observed in the blank control.
Toxic effects other than mortality, such as loss of coordination, hypo or hyperactivity and swimming on the back, were not observed.

Conclusions:

The 96-h LL50 value for Nourypol was based on the read across substance Sovermol 1055 and was determined to be above 100 mg/L based on loading (> 4.39 mg/l geometric mean concentration) in Danio rerio (Zebrafish) .

Executive summary:

The acute lethal toxicity of Nourypol to Danio rerio (Zebrafish) was read across from the substance Sovermol 1055. Sovermol 1055 was investigated in a guideline OECD TG203 study under GLP and static exposure conditions over a period of 96 h. The singly applied loading rate was 100 mg/l. HPLC analysis of the test substance concentration was conducted in the definitive test at the beginning and after 24, 48, 72 and 96 h of exposure. The measured concentrations were 4.39, 4.42, 4.34, 4.38 and 4.44 mg/l, respectively. The geometric mean concentration over a period of 0-96 h (GM0-96 h) was 4.39 mg/l (100% of initial dissolved concentration). At the loading rate of 100 mg/l, none of a total of seven fish died after 96 h of exposure. No mortality was observed in the blank control. Toxic effects other than mortality, e.g. loss of coordination, hypo- or hyperactivity and swimming on the back, were not observed. The no-observed-effect concentration (NOEC) was >=100 mg/l loading rate and >=4.4 mg/l geometric mean concentration. Based on the binominal theory, the absence of mortality provides at least 99% confidence that the LL50 is greater than the applied loading rate of 100 mg/l and > 4.4 mg/l when based on geometric mean concentration.

Description of key information

The acute lethal toxicity of Soybean oil, epoxidized, methyl ester, reaction products with propylene glycol to Danio rerio (Zebrafish) was read across from the substance Fatty acids, C16-18 and C18-unsatd., Me esters, epoxidized. Fatty acids, C16-18 and C18-unsatd., Me esters, epoxidized was investigated in a guideline OECD TG203 study under GLP and static exposure conditions over a period of 96 h. The singly applied loading rate was 100 mg/l. HPLC analysis of the test substance concentration was conducted in the definitive test at the beginning and after 24, 48, 72 and 96 h of exposure. The measured concentrations were 4.39, 4.42, 4.34, 4.38 and 4.44 mg/l, respectively. The geometric mean concentration over a period of 0-96 h (GM0-96 h) was 4.39 mg/l (100% of initial dissolved concentration). At the loading rate of 100 mg/l, none of a total of seven fish died after 96 h of exposure. No mortality was observed in the blank control. Toxic effects other than mortality, e.g. loss of coordination, hypo- or hyperactivity and swimming on the back, were not observed. The no-observed-effect concentration (NOEC) was >=100 mg/l loading rate and >=4.4 mg/l geometric mean concentration. Based on the binominal theory, the absence of mortality provides at least 99% confidence that the LL50 is greater than the applied loading rate of 100 mg/l and > 4.4 mg/l when based on geometric mean concentration.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The LL50 was > 100 mg/L, the max loading tested.