Dibutyl fumarate

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

According to OECD 210, : “Fish, Early-life Stage Toxicity Test”, Adopted July 17, 1992. In addition, the procedures were designed to meet the test methods of the following guideline and guidance document:• EPA Ecological Effects Test Guidelines, OPPTS 850.1400, Fish Early-Life Stage Test 'Public Draft', EPA 712-C-96-121, April 1996.• Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000.

Qualifier:

according to guideline

Guideline:

OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)

Deviations:

yes

Remarks:

1. Semi-static range-finding test: The temperature of the old medium at day 1 was 28°C. Evaluation: This did not affect the embryos. 2. Final test: Illumination has been continuous. Evaluation: Embryos and larvae were not affected.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Samples were taken on day 0, day 1, day 7, day 14, day 21, day 28 and day 35. Measured concentrations were almost always lower than target and not stable. It was therefore considered appropriate to calculate the mean measured concentrations in order to determine the effect parameters. The mean measured concentrations were calculated to be: 0.0074, 0.062, 0.23, 1.2 and 5.7 mg/l.

Vehicle:

no

Details on test solutions:

0.050, 0.16, 0.50, 1.6 and 5.0 mg/l Test medium without test substance or other additives (blank-control) and one control containing test medium with the additives used in the treatment of the stock solutions (DMF-control).

Test organisms (species):

Pimephales promelas

Details on test organisms:

Fathead minnow (Pimephales promelas, Teleostei Cyprinidae) Rafinesque, In house culture.

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

35 d

Hardness:

Total hardness ranged between 196 and 250 mg calcium carbonate per litre. This was according to the value described in the protocol, i.e. >140 mg/l as CaCO3.

Test temperature:

The average temperature in the control and the treatment groups varied between 23.1 and 25.4°C. The temperature continuously measured in the control vessels ranged between 23.3 and 25.7°C. This was within the range described in the Protocol: 25 ± 2°C.

pH:

Oxygen concentrations varied between 6.8 and 8.6 mg/l during the test period, which is >60% of oxygen saturation at 25°C as prescribed by the Protocol. .

Dissolved oxygen:

Oxygen concentrations varied between 6.8 and 8.6 mg/l during the test period, which is >60% of oxygen saturation at 25°C as prescribed by the Protocol.

Salinity:

Adjusted ISO medium, formulated using RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands) with the following composition:CaCl2.2H2O211.5 mg/lMgSO4.7H2O 88.8 mg/lNaHCO3 46.7 mg/lKCl 4.2 mg/l

Nominal and measured concentrations:

Nominal: 0.050, 0.16, 0.50, 1.6 and 5.0 mg/l Measured: 0.0074, 0.062, 0.23, 1.2, 5.7 mg/l

Details on test conditions:

The spawning tank is equipped with a substrate (pvc-tube), which enables collection of the fertilised eggs.Feeding brood stock: Frozen brine shrimp Nauplii and pelleted fish food (Cyprico Crumble Excellent (300-500 um), Coppens International bv, Helmond, The Netherlands).Time of fertilisation: Males and females were put together in spawning tanks and spawning started the following day approximately 1 to 2 hours after lights were switched on.

Reference substance (positive control):

yes

Remarks:

DMF

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

ca. 0.23 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

other: embryonic survival

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

ca. 0.062 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

other: larval survival

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

0.007 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

other: larval growth

Details on results:

Overall survival of fertilised eggs in the DMF-control was ≥66% until hatching was complete (i.e. 100% after 6 days of exposure). No significant effects on mortality of fish embryo’s and juvenile fish were observed at test concentrations up to and including 0.50 mg/l during the 7-day range-finding test. At the highest target concentration of 5.0 mg/l all embryos died within the first two days of exposure.Samples taken from all test concentrations before the start of the test showed that the analytical results were variable and could not be improved before the start of the test. Samples taken from the stock solutions at day 1 (fresh) and at days 2 and 4 (old) showed that the concentrations in the four highest stocks were in agreement with target (90-116%), while the concentration in the lowest stock was slightly to high at the start (123% of target) and decreased to 80% of target after 72 hours.

The pH varied between 7.4 and 7.8 during the test period, which was within the range described in the Protocol

Validity criteria fulfilled:

yes

Conclusions:

The results of the ELS test led to the following conclusions:1. Fumaric acid di-n-butyl ester did not induce any statistically significant effects on embryonic survival at concentrations up to and including 0.23 mg/l. Hence, the NOEC and LOEC for embryonic survival were 0.23 and 1.2 mg/l, respectively;2. Fumaric acid di-n-butyl ester did not induce any statistically significant effects on larval survival at concentrations up to and including 0.062 mg/l. Hence, the NOEC and LOEC for larval survival were 0.062 and 0.23 mg/l, respectively;3. Fumaric acid di-n-butyl ester did not induce any statistically significant effects on larval growth at concentrations up to and including 0.0074 mg/l. Hence, the NOEC and LOEC for larval growth were 0.0074 and 0.062 mg/l, respectively

Executive summary:

Fish early-life stage toxicity test with Fumaric acid di-n-butyl ester.

 

The study procedures described in this report were based on theOECD guidelines for Testing of Chemicals: Guideline No. 210, 1992. In addition,the procedures were designed to meet the test methods and validity criteria of theEPA Ecological Effects Test Guidelines, 'Public Draft', EPA 712-C-96-121, 1996 andthe OECD series on testing and assessment number 23, 2000.

 

The reproduction test was performed using a flow-through system with target concentrations of 0.050, 0.16, 0.50, 1.6 and 5.0 mg/l, which were based on the results of two preceding range-finding tests. A blank- and solvent-control (DMF) were also included.

 

In the flow-through system the dilution water was dosed separately from the test substance stock solutions into mixing vessels. Stock solutions in DMF were a factor of 10,000 higher than in the target concentrations. The dosed volumes and the dilution water were mixed under continuous stirring in the mixing vessels before entering the test vessels. The dosing was computer controlled, and the system was checked daily. The ELS testwas performed with three replicates containing 20 eggs per replicate for each concentration and controls.The test started by placing fertilised eggs in stainless steel test vessels. During the embryonic and larval phases, the fish were observed for effects on development, appearance and swimming behaviour. At the end of the test (day 35), the surviving fish were measured and weighed.

 

During the study, samples for analyses were taken at the start of exposure and on days 7, 14, 21, 28 and 35. Measured concentrations were almost always lower than target and not stable. It was therefore considered appropriate to calculate the mean measured concentrations in order to determine the effect parameters. The mean measured concentrations were calculated to be: 0.0074, 0.062, 0.23, 1.2 and 5.7 mg/l.

 

The study met the acceptability criteria prescribed by the protocol and was considered valid.

 

The results of the ELS test led to the following conclusions:

 

1.    Fumaric acid di-n-butyl ester did not induce any statistically significant effects on embryonic survival at concentrations up to and including 0.23 mg/l. Hence, the NOEC and LOEC for embryonic survival were 0.23 and 1.2 mg/l, respectively;

2.    Fumaric acid di-n-butyl ester did not induce any statistically significant effects on larval survival at concentrations up to and including 0.062 mg/l. Hence, the NOEC and LOEC for larval survival were 0.062 and 0.23 mg/l, respectively;

3.    Fumaric acid di-n-butyl ester did not induce any statistically significant effects on larval growth at concentrations up to and including 0.0074 mg/l. Hence, the NOEC and LOEC for larval growth were 0.0074 and 0.062 mg/l, respectively.

Description of key information

The results of the ELS test led to the following conclusions:

1. Fumaric acid di-n-butyl ester did not induce any statistically significant effects on embryonic survival at concentrations up to and including 0.23 mg/l. Hence, the NOEC and LOEC for embryonic survival were 0.23 and 1.2 mg/l, respectively; 2. Fumaric acid di-n-butyl ester did not induce any statistically significant effects on larval survival at concentrations up to and including 0.062 mg/l. Hence, the NOEC and LOEC for larval survival were 0.062 and 0.23 mg/l, respectively;

3. Fumaric acid di-n-butyl ester did not induce any statistically significant effects on larval growth at concentrations up to and including 0.0074 mg/l. Hence, the NOEC and LOEC for larval growth were 0.0074 and 0.062 mg/l, respectively

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

7.4 µg/L