Isooctadecanoic acid, mixed esters with oxybis[propanediol]

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

refer to Analogue Justification provided in IUCLID section 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

93.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mortality (fish)

Remarks on result:

other: Source: CAS 73296-86-3, monoester

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Source: CAS 67938-21-0

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

25 July - 1 Aug 1980

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test follows accepted Guidelines available in 1980, no GLP, limited but sufficient reporting

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

Test concentrations nominal: 0-1-10-100-500 mg/L

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Dispersion of the test item in test medium
- Controls: Blank control

Test organisms (species):

Leuciscus idus melanotus

Details on test organisms:

TEST ORGANISM
- Common name: Golden orfe
- Source: Paul Eggers, 2345 (now 24594) Hohenweststedt
- Length at study initiation (length definition, mean, range and SD): range 5.5-6.1 cm, mean 5.9 cm, SD 0.2 cm
- Weight at study initiation (mean and range, SD): range 1.4-2.5 g, mean 2.1 g, SD 0.32 g

ACCLIMATION
- Acclimation period: 14 days before test start
- Acclimation conditions (same as test or not): not same; acclimatisation in dechlorinated tap water at 20°C
- Type of food during acclimation: Tetra Min (Tetra Werke, Melle)

FEEDING DURING TEST
not specified

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

total hardness 9.5 dH
carbonate hardness 6.4 dH

Test temperature:

20 ± 1 °C

pH:

8.0 - 8.3

Dissolved oxygen:

7 mg/L

Nominal and measured concentrations:

nominal: blank-1-10-100-500 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass basin (40x25x30 cm), filled with 20 L of test medium
- Aeration: aeration through glass capillars; flow rate: 100 mL air/min
- 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: deionisied tap water (CHRIST-Ministil-Ionenaustauschpatrone), conductivity < 5 µS/cm; preparation of the test medium by adding 192 mg NaHCO3, 120 mg CaSO4 x 2 H2O, 120 mg MgSO4 and 8 mg KCl
- Intervals of water quality measurement: at test start and after 2, 24, 48, 72 and 96 hrs

OTHER TEST CONDITIONS
- Photoperiod: 12/12 dark/light
- Light intensity: 700 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and abnormal behaviour

TEST CONCENTRATIONS
- Test concentrations: Blank control, 1, 10, 100 and 500 mg/L

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Fish of the highest concentration (500 mg/L) swimming at the surface at 24 hrs after test start. All other groups showed no behaviour different to the control group. No macroscopically visible effects on the fish were observed in any group after euthanasia.

Sublethal observations / clinical signs:

Table 1: Validity criteria

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	0%	yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	>60%	yes

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 May - 26 June 1998

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

No analytics performed. No dose-response related mortality. Test concentrations were turbid and all concentrations tended to dehomogenise during exposure.

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

no analytics performed to demonstrate stability and allow for quantification of the test item throughout the study period

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

17 July 1992

Deviations:

yes

Remarks:

no analytics performed to demonstrate stability and allow for quantification of the test item throughout the study period

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preparation of water accomodated fractions (WAFs)
Since the test substance was insoluble in water, the toxicity of water accomodated fractions (WAFs) was tested. Magnetic stirring was applied for 2 days to achieve the maximum soluble fraction of the test item in the test medium. This solution was then transferred to a separating funnel and left to stabilise for 1 day. The water phase in the middle of the funnel was used for testing and identified as WAF. The appearance of the final test solutions ranged from clear and colourless (1 mg/L WAF) to turbid at the higher test concentration.
- Controls: blank control consisting of test medium only without the test substance or other additives

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Source: Bio International, Roermond, Netherlands
- Length at study initiation (length definition, mean, range and SD): First test: mean length 2.1 cm (SD 0.17 cm); second test: 2.15 cm mean length (SD 0.38 cm)
- Weight at study initiation (mean and range, SD): First test: mean weight 0.17 g (SD 0.04 g); second test: 0.42 g mean weight (SD 0.10 g))
- Method of breeding: F1 from a single parent-pair bred in UV-treated water

ACCLIMATION
- Acclimation period: at least 12 days after delivery
- Acclimation conditions (same as test or not): same as test medium
- Type and amount of food during acclimation: feeding with Trouvit
- Feeding frequency during acclimation: daily
- Health during acclimation (any mortality observed): mortality during the 7 days prior to test start was less than 5%

FEEDING DURING TEST
no feeding from 48 hours prior to test start and during the total test period

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

250 mg CaCO3

Test temperature:

20.4 - 21.8 °C

pH:

Day 0: 7.7 - 8.0
Day 1: 7.8
Day 2: 7.5 - 7.7
Day 3: 7.3 - 7.7
Day 4: 7.4 - 7.6

Dissolved oxygen:

Day 0: 8.4 - 9.0 mg/L
Day 1: 8.3 - 8.7 mg/L
Day 2: 7.5 - 7.9 mg/L
Day 3: 7.1 - 7.2 mg/L
Day 4: 7.0 - 7.1 mg/L

Nominal and measured concentrations:

nominal: control, 10, 100, 1000 and 10,000 mg/L and 10% of the WAF prepared at 10 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: depending on fish weight; 2, 3, 4 or 8 L all-glass vessels
- Aeration: introduced after 24 hours of exposure in the second test. No aeration in the first test.
- No. of organisms per vessel: 3 fish at 10, 100 and 1000 mgL (WAF) and to 10% of the WAF prepared at 10 mg/L (nominal); 7 fish at 10,000 mg/L (WAF, nominal) and blank control
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: FIRST TEST: 0.48 g fish per L, corresponding to 7 fish per 2.5 L of test medium (3 L vessel); 0.34 g fish per L, corresponding to 3 fish per 1.5 L of test medium (2 L vessel); SECOND TEST: 0.59 g fish per L, corresponding to 7 fish per 5 L of test medium (8 L vessel); 0.63 g fish per L, corresponding to 3 fish per 2 L of test medium (4 L vessel)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium formulated using Milli-Ro water (i.e. tap water purified by reverse osmosis); the following ions were added: Ca2+ 80 mg/L, Mg2+ 12 mg/L, Na+ 15 mg/L, K+ 3 mg/L, Cl- 145 mg/L, SO4 2- 49 mg/L, HCO3- 47 mg/L;
- Intervals of water quality measurement: dissolved oxygen concentration, pH and temperature measured daily beginning at day 0 (test start)

OTHER TEST CONDITIONS
- Photoperiod: 16 hrs photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, fish length and fish weight

TEST CONCENTRATIONS
Water Accomodated Fractions (WAFs) prepared at nominal 10, 100, 1000 and 10,000 mg/L and 10% of the WAF prepared at 10 mg/L
Blank Control prepared without the test item or any other additives

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol (PCP)

Dose descriptor:

other: No dose descriptor value could be determined.

Remarks on result:

not determinable

Remarks:

Results of both tests showed that mortality occured at random instead of dose-related. Test concentrations were increasingly turbid and all dispersions tended to dehomogenise during exposure. Substance particles were observed sticking to some of the fish. Hence, instead of toxicological effects, mechanical effects such as clogging of the gills may have caused the observed non dose-related mortality.

Details on results:

Results of both tests showed that mortality occured at random instead of dose-related. Test concentrations were increasingly turbid and all dispersions tended to dehomogenise during exposure. Substance particles were observed sticking to some of the fish. Hence, instead of toxicological effects, mechanical effects such as clogging of the gills may have caused the observed non dose-related mortality.

Results with reference substance (positive control):

Concentrations: Blank, 0.10, 0.15, 0.22, 0.32 and 0.46 mg/L in ISO medium
5 fish per treatment were exposed to the reference substance Pentachlorophenol (PCP) over a period of 96 hours.
LC50 (24h) = 0.15 mg/L
LC50 (96h) = 0.10 mg/L
The 96h LC50 was already reached after 48 hours of exposure.

Sublethal observations / clinical signs:

Table 1: Incidence and Cumulative mortality

Test substance/WAF [mg/L]	Initial number of fish	Cumulative mortality					Total mortality [%]
		2 h	24 h	48 h	72 h	96 h
Blank control	7	0	0	0	0	0	0
10% of WAF prepared at nominal 10 mg/L	3	0	0	0	0	0	0
10	3	0	0	0	1	3	100
100	3	0	0	0	1	2	67
1000	3	0	0	0	0	0	0
10,000	7	0	0	0^1)	1^2)	4^3)	57

1) All fish were swimming hypoactive compared to the fish in the control.

2) Six fish were swimming hypoactive compared to the fish in the control.

3) Two fish were swimming hypoactive compared to the fish in the control, while one fish was immobile.

Table 2) Validity criteria

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	0%	yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	>60%	yes

Validity criteria fulfilled:

yes

Remarks:

No dose-response related mortality. Test concentrations were turbid and all concentrations tended to dehomogenise during exposure. For further details please refer to "Any other information on results incl. tables."

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

11 May - 6 June 1998

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

GLP guideline study without analytics

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

17 July 1992

Deviations:

yes

Remarks:

no analytics performed to demonstrate stability and allow for quantification of the test item throughout the study period

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

no analytics performed to demonstrate stability and allow for quantification of the test item throughout the study period

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preparation of water accomodated fractions (WAFs)
Since the test substance was insoluble in water, the toxicity of water accomodated fractions (WAFs) was tested. Magnetic stirring was applied for 2 days to achieve the maximum soluble fraction of the test item in the test medium. This solution was then transferred to a separating funnel and left to stabilise for 1 day. The water phase in the middle of the funnel was used for testing and identified as WAF. The appearance of the final test solutions ranged from clear and colourless to turbid at the highest test concentration.
- Controls: blank control consisting of test medium only without the test substance or other additives

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Source: Zodiac, proefacc, "De Haar Vissen", L.U., Wageningen , Netherlands
- Length at study initiation (length definition, mean, range and SD): Combined limit/range finding tests: mean length 2.1 cm (SD 0.17 cm); final test: 2.07 cm mean length (SD 0.07 cm)
- Weight at study initiation (mean and range, SD): Combined limit/range finding tests: mean weight 0.17 g (SD 0.04 g); final test: 0.18 g mean weight (SD 0.03 g))
- Method of breeding: F1 from a single parent-pair bred in UV-treated water

ACCLIMATION
- Acclimation period: at least 12 days after delivery
- Acclimation conditions (same as test or not): same as test medium
- Type and amount of food during acclimation: feeding with Trouvit
- Feeding frequency during acclimation: daily
- Health during acclimation (any mortality observed): mortality during the 7 days prior to test start was less than 5%

FEEDING DURING TEST
no feeding from 48 hours prior to test start and during the total test period

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

250 mg CaCO3 per L

Test temperature:

20.1 - 20.3 °C

pH:

Day 0: 7.9 - 8.0
Day 1: 7.6
Day 2: 7.7 - 7.8
Day 3: 7.7 - 7.8
Day 4: 7.6 - 7.7

Dissolved oxygen:

Day 0: 8.6 - 9.1 mg/L
Day 1: 6.5 - 7.0 mg/L
Day 2: 7.9 - 8.3 mg/L
Day 3: 8.0 - 8.3 mg/L
Day 4: 8.0 - 8.1 mg/L

Nominal and measured concentrations:

nominal: control, 4.6, 10, 22, 46 and 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 2 L all-glass vessels
- Aeration: introduced after 24 hours of exposure
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.74 g fish per L, corresponding to 7 fish per 1.7 L of test medium

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium formulated using Milli-Ro water (i.e. tap water purified by reverse osmosis)
- Intervals of water quality measurement: dissolved oxygen concentration, pH and temperature measured daily beginning at day 0 (test start)

OTHER TEST CONDITIONS
- Photoperiod: 16 hrs photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, fish length and fish weight

TEST CONCENTRATIONS
- Range finding study : combined limit/range-finding study; 3 fish per concentration were exposed to a range of WAFs prepared at nominal 10 to 1000 mg/L with an increasing factor of 10 and to a 10-fold dilution of the 10 mg/L WAF. 7 fish were exposed to a blank control and a WAF prepared at 10,000 mg/L.
- Results used to determine the conditions for the definitive study: at the end of the test, all fish exposed to the WAF prepared at the nominal 100 mg/L had died. One fish exposed to the WAF at 10 mg/L had died. Based on these results, the 96h LC50 was expected to correspond with a WAF prepared at a nominal concentration between 10 and 100mg/L.

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol (PCP)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

136.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Water Accomodated Fraction (WAF)

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

93.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Water Accomodated Fraction (WAF)

Basis for effect:

mortality (fish)

Details on results:

no mortality was observed in the blank control group

Results with reference substance (positive control):

Concentrations: Blank, 0.10, 0.15, 0.22, 0.32 and 0.46 mg/L in ISO medium
5 fish per treatment were exposed to the reference substance Pentachlorophenol (PCP) over a period of 96 hours.
LC50 (24h) = 0.15 mg/L
LC50 (96h) = 0.10 mg/L
The 96h LC50 was already reached after 48 hours of exposure.

Sublethal observations / clinical signs:

Table 1: Incidence and Cumulative mortality

Test substance/WAF [mg/L]	Initial number of fish	Cumulative mortality					Total mortality [%]
		2 h	24 h	48 h	72 h	96 h
Blank control	7	0	0	0	0	0	0
4.6	7	0	0	0	0	0	0
10	7	0	0	0	0	0	0
22	7	0	0	0	0^2)	0^2)	0
46	7	0	0	0	0	0	0
100	7	0	0	0^1)	2^1)	4^1)	57

1) Two fish suffered from a loss of equilibrium.

2) One fish showed a loss of righting reflex after 72 hours of exposure and this fish became immobile at the end of the test period.

Table 2: Validity criteria

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	0%	yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	> 60%	yes

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to "Any other information on results incl. tables."

Additional information

Since no studies investigating the short-term toxicity of Isooctadecanoic acid, mixed esters with oxybis[propanediol] to fish are available, in accordance with Regulation (EC) No 1907/2006 Annex XI, 1.5, a read-across to the structurally related source substance Isooctadecanoic acid, ester with oxybis[propanediol] (CAS 73296-86-3) and Di(isooctadecanoic) acid, diester with oxydi(propanediol) (CAS 67938-21-0) was conducted.

The source substances are representative to evaluate the short-term toxicity of the target substance to fish.This read-across is justified in detail within the analogue justification in IUCLID Section 13.

The target substance Isooctadecanoic acid, mixed esters with oxybis[propanediol] exists as two different grades. Grade One is characterized by mainly triesters of C18 iso fatty acids of the alcohol component diglycerol. Grade Two is characterized by mainly tetraesters of C18 iso fatty acids of the alcohol component diglycerol.

The source substance Isooctadecanoic acid, ester with oxybis[propanediol] (CAS 73296-86-3) also exists as two different grades. It is characterized as a UVCB substance with mainly monoesters (Grade One) and mainly diesters, respectively (Grade Two), of C18 iso fatty acids of the alcohol component polyglycerol.

The source substance Di(isooctadecanoic) acid, diester with oxydi(propanediol) (CAS 67938-21-0) is characterized by the alcohol component diglycerol which is esterified by C18 iso fatty acids and consists only of diesters.

One study per grade is available for the source substance Isooctadecanoic acid, ester with oxybis[propanediol] (CAS 73296-86-3) which were performed in identical test designs. Both GLP-studies were performed according to OECD 203 and EU method C.1. Due to the low water solubility of the test material, WAFs ("water accommodated fractions") prepared with the test medium were used to prepare the five test material loading rates. Cyprinus carpio were exposed for 96 h in a static freshwater system. After 96 h, the mortality of the fish was assessed.

In the first study with Isooctadecanoic acid, ester with oxybis[propanediol] (CAS 73296-86-3; Grade One), loading rates of 4.6, 10, 22, 46 and 100 mg test material/L were tested. No mortality was observed in the treatment or the control group. Only in the highest loading rate (100 mg/L) 57% mortality was reported. An EC₅₀(96 h) of 93.8 mg/L was determined.

In the second study with Isooctadecanoic acid, ester with oxybis[propanediol] (CAS 73296-86-3; Grade Two), loading rates of 0.10, 0.15, 0.22, 0.32 and 0.46 mg/L were tested. Results showed that mortality occured at random instead of dose-related. Test concentrations were increasingly turbid and all dispersions tended to dehomogenise during exposure. Substance particles were observed sticking to some of the fish. Hence, instead of toxicological effects, mechanical effects such as clogging of the gills may have caused the observed non dose-related mortality. The study was therefore disregarded for the evaluation of the short-term toxicity to fish.

The study with source substance Di(isooctadecanoic) acid, diester with oxydi(propanediol) (CAS 67938-21-0) was performed according to OECD 203. The fish (Leuciscus idus melanotus) were exposed to five treatment solutions with concentrations of 0, 1, 10, 100 and 500 mg/L over a period of 96 h in a static freshwater system. After 96 h, the mortality of the fish was assessed. No mortality was observed in the control or treatment groups. Therefore, an LC₅₀of > 500 mg/L was derived.

Based on the results for the structurally related read-across substances (in accordance with Regulation (EC) No 1907/2006 Annex XI, 1.5) which are characterized by a similar ecotoxicological profile and comparable structure, it can be concluded that no toxicity of the target substance Isooctadecanoic acid, mixed esters with oxybis[propanediol] to fish is expected up to the limit of the water solubility (< 0.15 mg/L).