Reaction product of 3,5,5-trimethyl-hexanoic acid and 2-methylpropanoic acid and pentaerythritol

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 May - 07 July 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study was conducted in accordance with International guidelines and in accordance with GLP. All guideline validity criteria were met.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark.
- Stability under test conditions: Stability confirmed in saturated solution over 96 h.
- Solubility and stability of the test substance in the solvent/vehicle: No solvent used. Test solution prepared from a saturated solution (i.e. WAF preparation).
- Reactivity of the test substance with the test medium: Stability confirmed in saturated solution over 96 h.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: test sample and dilution water were mixed to prepare the nominal concentration of 110 mg/L. After stirring for 48 h, the middle layer was collected and filtered with a glass fiber filter by neutral filtration to prepare the test solution.
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: N/A
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material): Treatment solution prepared from a WAF.

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable): N/A

OTHER SPECIFICS: No

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1.8 mg/L
- Sampling method: Samples (50 mL) were taken from the test solutions and filtered through a pre-conditioned glass fibre filter (0.4 µm). The filtrate was partitioned against ethyl acetate (30 mL). the solvent fraction was concentrated to dryness using a rotary evaporator. The resulting residue was reconstituted in ethyl acetate (2 mO) prior to GC analysis.
- Sample storage conditions before analysis: Not reported

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A water accomodated fraction (WAF) was prepared by dispensing test item into dilution water at a nominal concnetration of 110 mg/L. The solution was stirred for 48 h after which time thesolution was left to stand/ settle before the middle fraction of the solution was decanted and filtered. The filtrate was used to prepare the treatment solutions.
- Eluate: Dechlorinated tap water
- Controls: Tap water only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): N/A
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): N/A
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Medaka (Oryzias latipes)
- Strain: -
- Source: Laboratory stock
- Age at study initiation (mean and range, SD): 6 months old
- Length at study initiation (length definition, mean, range and SD): 2 ± 1 cm
- Weight at study initiation (mean and range, SD): Not reported
- Method of breeding: Not reported
- Maintenance of the brood fish: Maintained in dechlorinated tap water under test conditions.

ACCLIMATION
- Acclimation period: 61 days
- Acclimation conditions (same as test or not): Yes
- Type and amount of food during acclimation: 2 % bodyweight/ fish/ day
- Feeding frequency during acclimation: Daily
- Health during acclimation (any mortality observed): 0 % mortality 7 days prior to test initiation.

QUARANTINE (wild caught)
- Duration: N/A
- Health/mortality: N/A

FEEDING DURING TEST
- Food type: No feeding 24 h before start of exposure period
- Amount: N/A
- Frequency: N/A

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

44 mg/L as CaCO3

Test temperature:

23.8 - 24.2 ºC

pH:

7.7 - 7.9

Dissolved oxygen:

8.2 - 8.5 mg/L

Salinity:

Not reported

Conductivity:

17 mS/m

Nominal and measured concentrations:

1.8 mg/L solution prepared from a 110 mg/L WAF

Details on test conditions:

TEST SYSTEM
- Test vessel: Fish tank
- Type (delete if not applicable): Closed
- Material, size, headspace, fill volume: Glass, 3 L capacity (diameter 16 cm x depth 17 cm), filled to 200 mL.
- Aeration: Gentle
- Type of flow-through (e.g. peristaltic or proportional diluter): N/A
- Renewal rate of test solution (frequency/flow rate): Test solution renewed in their entirety at 48 h timepoint (i.e. semi-static conditions).
- No. of organisms per vessel: 4
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): N/A

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated tap water
- Total organic carbon: 0.4 mg/L
- Particulate matter: < 1 mg/L
- Metals: Mercury <0.0005 mg/L; cadmium <0.001 mg/L; chromium <0.005 mg/L; lead <0.001 mg/L; iron <0.03 mg/L; copper <0.001 mg/L; cobalt < 0.001 mg/L; manganese <0.005 mg/L; zinc <0.1 mg/L; aluminium 0.02 mg/L; nickel < 0.001 mg/L; sodium 16 mg/L; magnesium 3.3 mg/L; silver <0.0001 mg/L
- Pesticides: 1,2-dichloropropane < 0.006 mg/L; chlorothalonil <0.005 mg/L; propyzamide <0.0008 mg/L; chloronitrofen <0.0001 mg/L; simazine <0.0003 mg/L; thiobencarb <0.002 mg/L; diazinon <0.0005 mg/L; isoxanthion < 0.0008 mg/L; fenitrothion <0.0003 mg/L; EPN <0.0006 mg/L; dichlorvos <0.001 mg/L; iprobenfos <0.0008 mg/L; PCB <0.0005 mg/L
- Chlorine: <0.02 mg/L
- Alkalinity: 35 mg/L
- Ca/mg ratio: Not reported
- Conductivity: 17 mS/m
- Salinity: Not reported
- Culture medium different from test medium: No
- Intervals of water quality measurement: At test start and end

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16:8 light: dark
- Light intensity: Not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality and sub-lethal effects were evaluated at 3, 24, 48, 72 and 96 h

VEHICLE CONTROL PERFORMED: No

RANGE-FINDING STUDY
- Test concentrations: Limit test
- Results used to determine the conditions for the definitive study: N/A, limit test

Reference substance (positive control):

yes

Remarks:

Copper sulfate tested periodically at the Test Facility (previous test was conducted 15 - 19 November 2010)

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 1.8 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: No effects observed
- Observations on body length and weight: No effects observed
- Other biological observations: No effects observed
- Mortality of control: 0 %
- Other adverse effects control: No effects observed
- Abnormal responses: No effects observed
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

- Results with reference substance valid: Yes
- Relevant effect levels: 96-h LC50 determined as 0.54 ± 0.20 mg/L. The value was within the normal historical range (n=46).

Reported statistics and error estimates:

No statistical analysis were performed as no effects were observed in the test.

Sublethal observations / clinical signs:

Table 1      Measured Test Item concentrations

 

Nominal Concentration (mg/L)	Measured Concentration (mg/L) (Percentage of initial concentration)
	0 h	48 Hours		96 h	Geometric mean
		Expired	Fresh
Control	nd	nd	nd	nd	-
110	2.2	1.6 (73)	1.7	1.8 (107)	1.8

nd: < 0.29 mg/L

 

Table 2      Cummulative mortality

 

Measured concentration (mg/L)	Cumulative mortality (%)
	3 h	24 h	48 h	72 h	96 h
Control	0	0	0	0	0
1.8	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

The 96 h LL50 to Medaka, under the conditions of this test was >1.8 mg/L. No sub-lethal effects were noted in any of the tested concentrations.

Executive summary:

OECD 203 (2011) - In a 96-h acute toxicity study, Medaka (Oryzias latipes) were exposed to Tetraesters of pentaerythritol with 2-methylpropanoic acid and 3,5,5-trimethyl-hexanoic acid at a measured concentration of 1.8 mg/L under semi-static conditions.  The test solutions were prepared from a water accommodated fraction (WAF) which contained the test item suspended in dilution water at a nominal loading rate of 110 mg/L. The WAF was stirred continuously for 48 h. The solubilized fraction was decanted and filtered to prepare the treatment solution for this limit test. The treatment solutions were replaced at the 48 h timepoint. The fish were checked for mortality and sub-lethal effects at 3, 24, 48, 72 and 96 hours after the start of the test item exposure.

 

No mortality or sub-lethal effects were observed during the test. The resulting 96-h LL₅₀ and NOEL were therefore determined to be >1.8 mg/L and 1.8 mg/L, respectively.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for acute toxicity in fish.

 

Description of key information

96 h LL50 = >1.8 mg/L; OECD 203; Matsuura, T. (2011)

Additional information

OECD 203 (2011) - In a 96-h acute toxicity study, Medaka (Oryzias latipes) were exposed toTetraesters of pentaerythritol with 2-methylpropanoic acid and 3,5,5-trimethyl-hexanoic acid at a measured concentration of 1.8 mg/L under semi-static conditions.  The test solutions were prepared from a water accommodated fraction (WAF) which contained the test item suspended in dilution water at a nominal loading rate of 110 mg/L. The WAF was stirred continuously for 48 h. The solubilized fraction was decanted and filtered to prepare the treatment solution for this limit test. The treatment solutions were replaced at the 48 h timepoint. The fish were checked for mortality and sub-lethal effects at 3, 24, 48, 72 and 96 hours after the start of the test item exposure.

 

No mortality or sub-lethal effects were observed during the test. The resulting 96-h LL₅₀ and NOEL were therefore determined to be >1.8 mg/L and 1.8 mg/L, respectively.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for acute toxicity in fish.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for acute toxicity in fish.