Reaction mass of 2,2,4-trimethylhexane-1,6-diol and 2,4,4-trimethylhexane-1,6-diol

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:

2016-03-14 to 2016-03-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: (7) Series on Testing and Assessment, No. 126, Short Guidance on the Threshold Approach for Acute Fish Toxicity

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

The limit concentration and the control were analytically verified the test item via LC-MS/MS from freshly prepared media at the start and the end of the exposure.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
An appropriate volume of the test item was pipetted directly into aquaria filled with an appropriate volume of the dilution water. The test media was mixed with an ultraturrax (1 min, 17000 rpm).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

- Supplier: Umweltbundesamt, Schichauweg 58, D-12307 Berlinj
- holding at the test facility at 23 ± 2°C and diffuse light (0.1 - 10 µmol photons · m-2 · s-1, natural photoperiod). The water was changed at least once per week.
- at least 12 days of acclimatisation and mortality < 5 % within these days before the start of exposure
- Feeding: food was provided 3 times per week (Sera Vipan; SERA GMBH). The amount of food was 4 % of the fish body weight per feeding day. The test fish were not be fed 24 h before the test start.
- Water: Tap water of local origin. The water was filtered on activated charcoal and aerated for at least 24 hours to remove chlorine.
Nominal water parameters: Hardness: 10 - 250 mg CaCO3/L; pH-value: 6.0 - 8.5

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

23 ± 2°C

Dissolved oxygen:

not less than 60% of air saturation

Nominal and measured concentrations:

Nominal: 26 mg/L, tested as a threshold concentration

Details on test conditions:

- Preparation of the test medium: an appropriate volume of the test item was pipetted directly into aquaria filled with an appropriate volume of the dilution water. The test media was mixed with an ultraturrax (1 min, 17000 rpm).
- Control: 7 fish in dilution water (without test item) were tested under the same test conditions as the test replicates.
- Application: one application at the start of the study was performed.
- Test vessel : Glass aquaria of 13 L, filled with approximately 10 L (dimensions: 25.5/18/28 cm) , covered with glass plates.
- Test volume: about 10 L per vessel
- Aeration: gentle aeration
_ Dilution water: same as used for holding
- Number of fish: 7 zebrafish were used for each test concentration and the control (14 zebrafish for the whole study).
- Loading: fish density = 0.132 g fish per litre test solution.
- Average body length at the test end: 2.54 cm
- Average body weight at the test end: 0.189 g
- Replicates: one replicate per test concentration and control
- Introduction of fish: fish were introduced randomly to individual replicates.
- Feeding of test fish: the fish were not fed during the test.
- Light intensity: 0.1 – 10 µmol photons · m-2 · s-1 (corresponding to approximately 7 – 750 Lux)
- Photoperiod daily 12 hours photoperiod, which corresponded to the natural daylight hours, was maintained throughout exposure.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Observation intervalls: after 2, 24, 48, 72 and 96 h.
- Criteria of effects: fish were considered dead if there was no visible movement (e.g. gill covers movement) and if touching of the caudal peduncle produced no reaction. Records were kept of visible abnormalities (e.g. loss of equilibrium, swimming behaviour, respiratory function, pigmentation, etc.).
- Measurement of fish size ofcontrol fish at the end of exposure.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 26 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Results with reference substance (positive control):

No reference substance

Sublethal observations / clinical signs:

Measured Exposure Concentrations during the Definitive Test

The measured concentration of the test item at the start of the exposure (0 hours) was 28.8 mg/L. At the end of the exposure (96 hours), the measured concentration of the test item was 28.8 mg/L.

 

Table6:   Measured Concentrations and Percentage of the nominal Concentration of the Test Item

Sampling date		2016-03-14 (0 hours)			2016-03-18 (96 hours)
Date of Analysis		2016-03-14			2016-03-18
		2,2,4‑(or 2,4,4)‑Trimethylhexane‑1,6‑diol
Nominal test item concentration [mg/L]		Measured conc. [mg/L]		[%]		Measured conc. [mg/L]		[%]
26.0		28.8		111		28.8		111
Control		< LOQ			< LOQ

Measured conc.= Measured concentration of the test item, dilution factor taken into account

% = Percentage of nominal concentration of the test item

LOQ = Limit of quantification (60.0 µg/L test item)

Validity criteria fulfilled:

yes

Conclusions:

In this study, the test substance was found to have no effect on the vitality of zebrafish after 96 hours at a nominal test item concentration of 26.0 mg/L.
In conclusion, the LC0 corresponds to the nominal test item concentration ≥ 26.0 mg/L. The LC50 and the LC100 correspond to the nominal test item concentration > 26.0 mg/L.

Executive summary:

The acute toxicity of the test substance to fish (zebrafish) was determined according to the principles of the OECD-Guideline for Testing of Chemicals No. 203 (1992) and EU Council Regulation (EC) No. 440/2008/ Method C.1 (2008) from 2016-03-14 to 2016-03-18 at the test facility.

An acute toxicity test under static conditions with the nominal threshold concentration of 26 mg/L was carried out. The selection of the test concentration was based on the derivation of a threshold concentration (TC) from the results of a daphnia toxicity test (ErC50 (0-48 h)=80 mg/L (provided by the sponsor) and an algae acute toxicity test (EyC50 (0 – 72 h)= 26.1 mg/L. The duration of the test was 96 hours. Seven test organisms were exposed to each test concentration and the control. The water quality parameters (temperature, pH-value and oxygen saturation) measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits.

The determination of the test concentrations was carried out via LC-MS/MS. Evaluation was done from freshly prepared media after 0 h and from the corresponding aged test media after 96 h.

The measured concentration of the test substance in freshly prepared media at test start was 111% of the nominal value. The measured concentration at the end of exposure in the corresponding aged test media was 111% of the nominal value. Therefore, all effect values were based on nominal test item concentrations. The LC50 and the LC100 correspond to the nominal test item concentration > 26.0 mg/L.

Description of key information

The acute toxicity of the test substance to fish (zebrafish) was determined according to the principles of the OECD-Guideline for Testing of Chemicals No. 203 (1992) and EU Council Regulation (EC) No. 440/2008/ Method C.1 (2008) from 2016-03-14 to 2016-03-18 at the test facility.

An acute toxicity test under static conditions with the nominal threshold concentration of 26 mg/L was carried out. The selection of the test concentration was based on the derivation of a threshold concentration (TC) from the results of a daphnia toxicity test (EC50 (0-48 h)=80 mg/L (provided by the sponsor) and an algae acute toxicity test (EYC50 (0 – 72 h)= 26.1 mg/L. The duration of the test was 96 hours. Seven test organisms were exposed to each test concentration and the control.

The test substance was found to have no effect on the vitality of zebrafish after 96 hours at a nominal test item concentration of 26.0 mg/L.

In conclusion, the LC0 corresponds to the nominal test item concentration ≥ 26.0 mg/L. The LC50 and the LC100 correspond to the nominal test item concentration > 26.0 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

26 mg/L