Diesters of alcohols, C7-9-iso-, C8-rich, 2-ethylhexyl and nonanedioic acid

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:

04 - 08 May 1998

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study with acceptable restrictions: No analytics of the test concentrations performed. Analytical purity not stated.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

adopted in 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

adopted in 1992

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Preparation started with stock solutions prepared at desired nominal concentrations applying magnetic stirring for ~ 48 h to achieve the maximum soluble fraction of the test substance in the test medium. Subsequently, these stock solutions were transferred to a separating funnel and left to stabilize for ~ 26 h. The water phase in the middle of the separating funnel was used for testing.
- Evidence of undissolved material: The appearance of the final test solutions ranged from clear and colourless to turbid at the higher test concentrations.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: carp
- Source: Bio International, Roermond, The Netherlands
- Length at study initiation: 2.01 ± 0.11 cm
- Weight at study initiation: 0.18 ± 0.03 g
- Method of breeding: F1 from a single parent-pair bred in UV-treated water.
- Feeding during test: no

ACCLIMATION
- Acclimation period: at least 12 days after delivery
- Acclimation conditions: same as test
- Type and amount of food: Trouvit
- Feeding frequency: daily
- Health during acclimation: In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

250 mg/L as CaCO3

Test temperature:

20.0 - 20.6

pH:

7.3 - 8.1

Dissolved oxygen:

6.7 - 9.2 mg/L (in the 1000 mg/L treatment a much lower oxygen concentration of 3.2 mg/L was measured on day 2)

Nominal and measured concentrations:

Nominal: 0.1 x 10, 10, 100, 1000, 10000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 3 L vessels for 0.1 x 10, 10, 100 and 1000 mg/L treatments; 4L vessels for blank and 10000 mg/L treatment
- Material: glass
- Aeration: The medium was aerated until the dissolved oxygen concentration had reached saturation followed by aeration after 48 h of exposure
- No. of organisms per vessel: 3 in 3 L vessels and 7 in 4 L vessels
- No. of vessels per concentration: 1
- No. of vessels per control: 1
- Biomass loading rate: 0.5 g/L in 3 L vessel; 0.36 g/L in 4 L vessels

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water purified with reverse osmosis (Millipore Corp., Bedford, Mass., USA)
- Ca/mg ratio: 6.6
- Culture medium different from test medium: no
- Intervals of water quality measurement: The temperature was measured daily in the blank-control vessel. The pH was measured daily in the blank-control vessel and the highest concentration. Dissolved oxygen was measured daily in all vessels.

OTHER TEST CONDITIONS
- Photoperiod: 16 h photoperiod daily

EFFECT PARAMETERS MEASURED: Mortality was recorded 2, 24, 48, 72 and 96 hours following the start of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Range finding study: A range finding test was combined with a limit test as no lethal effects in fish were expected. However, the test substance was insoluble in water according to the information supplied by the sponsor. Therefore, a pretest was performed to examine the solubility of the test substance in Milli-Q water.
- Results used to determine the conditions for the definitive study: Since test substance was floating on the surface in a stock solution prepared at 12.5 g/L, it was decided to test toxicity of Water Accomodated Fractions (WAF).

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

other: siphoned test solution

Basis for effect:

mortality (fish)

Details on results:

- Abnormal responses: Any clinical abnormalities observed during the test are listed in Table 1.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: see Table 2

Results with reference substance (positive control):

The estimated LC50 of this test decreased from 0.15 mg/L after 24 h of exposure to 0.1 mg/L at the end of the test period. The 96 h LC50 was already reached after 48 h. The range of the 96 h LC50 for carp is generally between 0.1 and 0.46 mg/L based on historical data of reference tests conducted ~ every 3 months since April 1988. The carp originating from this batch can be considered as sensitive to toxic substances.

Sublethal observations / clinical signs:

Table 1: Clinical effects observed during the study

Treatment [mg/L]	Recording time [h]	Specification of effects	Relative Number
Control	0-48, 72-96	No abnormalities	7/7, 6/6
0.1x10, 10	0-96	No abnormalities	3/3
100	0-24	No abnormalities	3/3
	48	Hypoactive swimming	3/3
		Haemorrhage of the tail	1/3
	72	Loss of equilibrium and
		haemorrhage of the gill	1/3
	96	Immobile and haemorrhage
		of the gills	1/3
1000	0-24, 72	No abnormalities	3/3
	48	Hypoactive swimming	3/3
	96	Loss of equilibrium	1/3
10000	0-2	No abnormalities	7/7
	24-72, 96	Hypoactive swimming	7/7, 6/7
	24, 48	Swimming at the surface	7/7, 3/7
	48	Swimming at the bottom	4/7
	96	Loss of equilibrium	1/7

Table 2: Incidences and total mortality

Treatment [mg/L]	Initial number of fish	Cumulative mortality					Total Mortality [%]
		2 h	24 h	48 h	72 h	96 h
Control	7	0	0	0	1	1	14
0.1x10	3	0	0	0	0	0	0
10	3	0	0a	0a	0	0	0
100	3	0	0a	0a	0	0	0
1000	3	0	0a	0a	0	0	0
10000	7	0	0a	0a	0a	0a	0

a: Test substance film on the surface

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 - 22 Oct 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions: Analytical purity not stated.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test solution was prepared according to the national standard procedure (DEV S4): 10 g of the test substance was added to 1 L deionised water and shacked overhead for 24 h. The solution was filtered with a folded filter. The filtered solution was added directly to the test vessels.

Test organisms (species):

Leuciscus idus melanotus

Details on test organisms:

TEST ORGANISM
- Common name: golden orfe
- Source: Fischzucht Eggers, Hohenwestedt, Germany
- Age at study initiation: approx. 4 weeks
- Corpulence factor: 0.8 - 1.1 g/cm3


ACCLIMATION
- Acclimation conditions (same as test): yes
- Type and amount of food: food directly from the supplier of the animals
- Feeding frequency: 2 times a week

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

15 °dH

Test temperature:

20 ± 1 °C

pH:

8.3 - 8.6

Dissolved oxygen:

7.4 - 9.2 mg/L

Nominal and measured concentrations:

Nominal concentration: 10000 mg/L filtered test solution

Details on test conditions:

TEST SYSTEM
- Material, size, fill volume: glass, 28.5x21.0x24.0 cm, 10 L
- Aeration: slightly aerated
- No. of organisms per vessel: 10
- No. of vessels per concentration: 1
- No. of vessels per control: 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Drinking water from Cologne, Germany mixed with deionised water.

EFFECT PARAMETERS MEASURED: Mortality was recorded after 24, 48, 72, 96 h test duration.

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: Because the test substance is poorly water soluble the filtrate from a stock solution of 10 g/L was used to analyse if there are any effects in the range of water solubility.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

10 g/L

Nominal / measured:

nominal

Conc. based on:

other: filtered test solution

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 g/L

Nominal / measured:

nominal

Conc. based on:

other: filtered test solution

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%

Sublethal observations / clinical signs:

No effects of the test substance in the range of water solubility were observed testing a saturated test solution which was subsequently filtered. Thus, an LC50 (96h) greater then the highest tested concentration of 10 g/L is determined.

Description of key information

In a test in L. idus melatonus, no effects of C9-11 branched alcohols, C10 rich diesters with nonanedioic acid in the range of water solubility were observed testing a saturated test solution which was subsequently filtered. Thus, an LC50 (96h) greater than the highest tested concentration of 10 g/L is determined (Hansonis 1993)

In a test on bis(2-ethylhexyl) azelate, where carp were exposed to this substance during 96 hours, no mortality was observed at loading rates up to 10000 mg/L. Effects on swimming behaviour became apparent at concentrations of 100 mg/L and above. The LD50 based on loading rate is > 10000 mg/L (Bogers 1998)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

10 g/L

Additional information

No analytical measurements were included in the tests with both analogues. The results are, however, indicative for no mortality and very low toxicity at the water solubility level. As both substances are considered close analogues to diesters of alcohols, C7-9-iso-, C8-rich, 2-ethylhexyl and nonanedioic acid, no toxicity to fish at the water solubility level is expected for this substance. Therefore the LD50 is set at >10 g/L (see also document on read-across rationale)