COASOL 290 PLUS

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:

October 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name: COASOL 290 PLUS
Chemical name: Reaction mass of dibutyl adipate and dibutyl glutarate
Batch/Lot Number: 6H04163A01
Description: Clear colourless liquid
Purity: 99.3 %
Expiry date: 04 August 2019
Storage condition: Room temperature (15-25 °C, ≤ 70 RH%)
Safety precautions: Routine safety precautions (gloves, goggles, face mask, lab coat) for unknown materials were applied to assure personnel health and safety.

Analytical monitoring:

yes

Details on sampling:

Samples were taken at all concentrations as well as from the control, at the beginning and at the end of the first and at the last renewal periods.

Vehicle:

no

Details on test solutions:

Because the test item is a reaction mass and poorly soluble in test media, test solutions were prepared using a saturated solution method (water accommodated fraction, WAF) according to the Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, OECD No. 23 (2000). Test item solutions (nominal loading rates of 6.25, 12.5, 25.0, 50.0 and 100.0 mg/L) were prepared individually by dispersing/dissolving the amount of test item into the test medium (aquarium water) two days before the start of the experiment. These solutions were shaken for about 24 hours at approximately 30°C and then equilibrated for about 24 hours at approximately 20°C. The non-dissolved test material was removed by filtration through a fine (0.22 µm) filter to give the appropriate WAF solutions. The test solutions were prepared just before introduction of the fish (start of the treatments). Prior to treatment of each renewal period, test item solution was prepared by the method described above.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Zebrafish (Brachydanio rerio)
Source: István Szent University, 2100 Gödöllő, Páter Károly u. 1, Hungary
Justification of species: Zebrafish (Brachydanio rerio) is one of the convenient species for acute fish toxicity test.
Number of animals: seven fish per test group
Body length of animals: 2.5 – 2.9 cm
Food and Feeding: The fish were not fed during the test
Acclimatisation: more than 12 days
Animal health: Fish were bred in a well-known fish farm, under disease- and parasite-controlled conditions. Fish were observed during the acclimatisation and test period; the fish were healthy. There was no mortality of the population for seven days before the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

The hardness of the dilution water was determined as 187 mg/L (as CaCO3).

Test temperature:

The test temperature was between 20.0 and 22.0 °C.

pH:

The pH was in the range of 7.75 – 8.34.

Dissolved oxygen:

The dissolved oxygen concentration was in the range of 62 – 100 % of the air saturation value at the temperature used. Test solutions were aerated during the test to avoid decreasing of dissolved oxygen concentration below the accepted level.

Nominal and measured concentrations:

The nominal concentrations of test item used in the main experiment were: 6.25, 12.5, 25.0, 50.0 and 100.0 mg/L nominal loading rates WAFs.

Details on test conditions:

The test duration was 96 hours. One aquarium was used for test groups and for the control group, respectively. Each aquarium comprised 7 fish and 5 L test solution. The animals were not fed during the test. The loading of the test aquaria was less than 1.0 g fish/L test solution at the start of the experiment. The test was performed under semi-static conditions. The frequency of the water renewal periods was 24 hours. The choice of the test concentrations was made on the basis of the results of the preliminary range-finding test. In the range-finding test, four fish were exposed to each concentration of the test item plus a control, for 96 hours under semi-static conditions. The frequency of the water renewal periods was 24 hours. The concentration levels used and results (96 h) of the preliminary range-finding test are summarized in the following table:

Nominal concentrations
[mg/L nominal loading rate WAF] Untreated control 0.1 1 10 100
Number of treated animals 4 4 4 4 4
Number of dead animals 0 0 0 0 4

Because a toxic effect was observed at the highest concentration level during the preliminary range-finding test, five test concentrations and one untreated control were tested in the main experiment under semi-static conditions. The nominal concentrations of test item used in the main experiment were: 6.25, 12.5, 25.0, 50.0 and 100.0 mg/L nominal loading rates WAFs.

Reference substance (positive control):

not required

Key result

Duration:

24 h

Dose descriptor:

LL50

Effect conc.:

35.01 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

48 h

Dose descriptor:

LL50

Effect conc.:

35.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

35.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LL100

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOELR

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

yes

Executive summary:

The acute toxicity of COASOL 290 PLUS was assessed in an acute fish toxicity test on Zebrafish (Brachydanio rerio), over an exposure period of 96 hours in a semi-static system. Because a toxic effect was observed at the highest concentration level during the preliminary range-finding test, five test concentrations one untreated control were tested in the main experiment under semi-static conditions. The nominal concentrations of test item used in the main experiment were: 6.25, 12.5, 25.0, 50.0 and 100.0 mg/L nominal loading rates WAFs. The test item was supplied as a reaction mass of dibutyl adipate and dibutyl glutarate. The test item was not fully soluble at all tested concentrations, hence a WAF method was used to produce a range of concentrations at nominal loading rates from 6.25 to 100.0 mg/L. As is typical for WAF studies with substances described as a ‘reaction mass’ it was not known in advance what fractions of the supplied substance may be soluble or may be toxic, hence a general analytical method was used to measure the total organic carbon per mL aquatic media (TOC). The biological results are reported in units of the nominal loading rates; the TOC results are considered to be supporting data, confirming exposure. The biological results are based on the nominal loading rates WAFs. One aquarium was used in each test group and one in the control group. Each aquarium comprised seven fish and five litre test solution. The 24, 48, 72 and 96 hours LL50 values of the test item were calculated using Probit analysis by SPSS PC+ software. The NOELR and LOELR and LL100 values were determined from the raw data. All achievable validity criteria were met during this study.

 

Under the conditions of this acute fish toxicity studyon Zebrafish (Brachydanio rerio) the observed and calculated endpoints for the effect of COASOL 290 PLUS were the following:

Based on the nominal loading rates (WAFs):

The 24, 48, 72 and 96h LL₅₀value:                       35.01 mg/L (95 % conf. limits: 25.25 – 52.05 mg/L)

The 96h LL₁₀₀value:                                             50.0 mg/L

The 96h No-Observed Effect Loading Rate (NOELR):    25.0 mg/L

The 96h Lowest Observed Effect Loading Rate (LOELR):  50.0 mg/L

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

supporting study

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:

Marine fish species used

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
None

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1.6 mg/L WAF loading rate
- Sampling method: Samples were collected at 0 (fresh media), 24 (old media), 72 (fresh media) and 96 hours (old media) for analysis
- Sample storage conditions before analysis: Duplicate samples stored at -20 degrees Celsius for furhter analysis if necessary.

Vehicle:

no

Details on test solutions:

The 1.6 mg/L loading rate WAF was prepared by adding 33.6 mg of test material to the surface of 21 litres of synthetic sea water. This was stirred for 23 hours and then allowed to stand for 1 hour. A wide bore glass tube covered at one end with Nescofilm was then submerged in the vessel, sealed end down, to a depth of approximately 5cm from the bottom. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. The aqueous phase of the WAF was then removed by mid-depth syphoning (the firs 75 -100ml discarded). Microscopic inspection of the WAF showed no microdispersions or undissolved test material to be present.

Test organisms (species):

other: Scophthalmus maximus (juvenile turbot)

Details on test organisms:

Fish (Scopthalmus maximus) supplied by Viking Fish Farms Ltd, Ardtoe Marine laboratory (UK) were maintained in a glass tank with an activated carbon recirculation filter system. They were acclimatised to test conditions for 12 days. The lighting cycle was 16 hours of light and 8 hours of darkness with a 20 minute dawn/dusk transition. Water temperature was controlled at 14 degrees Celcius with a dissolved O2 content of greater than or equal to 8.0 mg/L (recorded daily).

The fish were fed commercial turbot pellets until 24 hours before the start of the test. There was no mortality during the acclimatisation period in the 7 days prior to the test. Fish length and weight were 5.1cm (SD 0.2) and 3.65g (SD0.35) respectively. The loading rate was therefore 1.3g bodyweight per litre.

Test type:

semi-static

Water media type:

saltwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

Not applicable

Hardness:

no data

Test temperature:

13-14 degrees Celsius

pH:

7.9-8.7

Dissolved oxygen:

7.5-8.7

Salinity:

31 o/oo

Nominal and measured concentrations:

Nominal: 1.6 mg/L loading rate water-accommodated fraction (WAF).

Chemical analysis of the fresh media at 0 and 72 hours showed measured concentrations of 0.391 and 1.23 mg/L, respectively. Analysis of the old media at 24 and 96 hours showed measured concentrations to be less than the limit of quantitation of 0.052 mg/L. The decline in measured concentrations was in line with stability analysis conducted over a 24-hour period. However, given that the test material was prepared as a WAF and that toxicity cannot be attributed to a single component or a mixture of components but to the test material as a whole, the results were based on nominal loading rates.

Details on test conditions:

In the definitive study, 20 L glass vessels were used for the control and test concentration. 7 fish were allocated at random to each vessel. The vessels were covered to reduce evaporation and maintained at a temperature of 14 degrees Celcius in a room with a 16/8 h light dark cycle for a period of 96 hours. Aeration of test vessels was acheived using a narrow bore glass tube inserted into each vessel. Fish were not fed during the study.

The test preparations were renewed daily (semi static) to ensure the test material remained near the nominal level and to prevent build up of nitrogenous waste products. Mortalities and sub lethal effects were recorded at 3, 6, 24, 48, 72 and 96 hours. Death was considered to be the absence of both respiratory movement and response to physical stimulation. Water temperature, pH and dissolved O2 were recorded daily.

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1.6 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Nominal WAF Loading Rate

Basis for effect:

other: mortality and sublethal effects

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 1.6 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Nominal WAF Loading Rate

Basis for effect:

mortality (fish)

Details on results:

During the study there were no mortalities and no sub-lethal effects.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not applicable

Sublethal observations / clinical signs:

The 23 hour mixing period for preparation of the WAF was considered sufficient and thus used in the preparation of the definitive test concentrations.

During the study there were no mortalities and no sub-lethal effects.

Validity criteria fulfilled:

yes

Conclusions:

The LL50 for the test material was greater than 1.6 mg/L WAF.

Executive summary:

A study was performed to assess the acute toxicity of COASOL to juvenile turbot (Scophthalmus maximus). The method followed OECD Test Guideline No. 203, EC Method C.1, and OSLO Paris Commission (OSPARCOM) Guidelines 1995.

 

Following an acute toxicity to Acartia tonsat est and marine algal inhibition test for the same test material, which showed the lowest EC50 values to be 1.6 mg/L loading rate water-accommodated fraction (WAF), a single group of seven fish were exposed to WAFs of the test material at a single nominal loading rate of 1.6 mg/L for a period of 96 hours at a temperature of ~14°C under semi-static (daily renewal) conditions.

 

Chemical analysis of the fresh media at 0 and 72 hours showed measured concentrations of 0.391 and 1.23 mg/L, respectively. Analysis of the old media at 24 and 96 hours showed measured concentrations to be less than the limit of quantitation of 0.052 mg/L. The decline in measured concentrations was in line with stability analysis conducted over a 24-hour period. However, given that the test material was prepared as a WAF and that toxicity cannot be attributed to a single component or a mixture of components but to the test material as a whole, the results were based on nominal loading rates.

 

The 96-hour LL50 was greater than 1.6 mg/L and correspondingly, the NOELR was 1.6 mg/L.

Description of key information

COASOL 290 PLUS is a reaction mass of dibutyl glutarate and dibutyl adipate. The acute toxicity of COASOL 290 PLUS was assessed in an acute fish toxicity test with Zebrafish (Brachydanio rerio), over an exposure period of 96 hours in a semi-static system. The test item was not fully soluble at all tested concentrations, hence a WAF method was used to produce a range of concentrations at nominal loading rates from 6.25 to 100.0 mg/L. The 24, 48, 72 and 96h LL₅₀values were 35.01 mg/L (95 % conf. limits: 25.25 – 52.05 mg/L).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

35.01 mg/L

Marine water fish

Marine water fish

Effect concentration:

1.6 mg/L

Additional information

One short-term marine fish study was available for the analogue substance COASOL. The study was conducted with the juvenile turbot (Scopthalmus maximus) and found to be of good quality and assigned a Klimisch score of one. Due to the low water solubility of the test substance, fish were exposed to the water-ammodated fraction resulting from a solution prepared at a nominal loading rate of 1.6 mg/L. The study was conducted for 96-hours, under semi-static conditions, following OECD Test Guideline 203, and under GLPs. No fish mortality or sublethal effects were observed during the exposure and the study concluded the 96-hour LL50 value was >1.6 mg/L.