Reaction mass of Terpenes and terpenoids, turpentine-oil, alpha-pinene fraction, dimers and Terpenes and terpenoids, turpentine-oil, alpha-pinene fraction, trimers

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

On one occasion the temperature was above than the range given in the guideline, but this was not considered to have affected the results of the test

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

yes

Remarks:

On one occasion the temperature was above than the range given in the guideline, but this was not considered to have affected the results of the test

Principles of method if other than guideline:

Due to the low aqueous solubility and complex nature of the test item, the test medium was prepared as a Water Accommodated Fraction (WAF) of the test item as recommended by regulatory authorities in the EU and elsewhere (ECETOC 1996, OECD 2000 and Singer et al. 2000).

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Test Material: Terpenes and terpenoids, turpentine oil, alpha-pinene fraction oligomers
CAS: 70750-57-1
EC: 500-245-8
Description: Extremely viscous pale amber colored liquid
Batch: 0910002624
Expiry date: Indefinite
Storage: Room temperature

Analytical monitoring:

yes

Details on sampling:

- Concentration: 100 mg/L loading rate WAF
- Sampling: water samples were taken from the control and the 100 mg/L loading rate WAF test group (replicates R1-R4 pooled) at 0 (fresh media), 24 (old and fresh media) and 48 hours (old media)
- Sample storage conditions before analysis: samples were stored frozen prior to analysis

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: the test medium was prepared as a Water Accommodated Fraction (WAF) of the test item. An amount of test item (2000 mg) was added to a glass slide and suspended in the water column of 20 L of test water to give the 100 mg/L loading rate. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 h and the mixture allowed to stand for 1 h. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the 100 mg/L loading rate WAF.

- Controls: negative control (ISO medium without test item), positive control (potassium dichromate)
- Evidence of undissolved material: Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present. After siphoning and for the duration of the test, the 100 mg/L loading rate was observed to be a clear, colorless solution.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Strain: Not reported
- Source: in-house laboratory culture
- Age at study initiation (mean and range, SD): <24 hours old
- Method of breeding: Parthenogenesis
- Feeding during test: None

ACCLIMATION
- Acclimation period: Yes
- Acclimation conditions (same as test or not): Adult Daphnia were kept in 150 mL glass beakers containing Elendt M7 medium in a temperature controlled room at approximately 20 °C.
- Type and amount of food: Daphnids were fed with mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension
- Feeding frequency: Daily

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

250 mg/L as CaCO3

Test temperature:

22 - 24°C

pH:

7.6 - 8.0

Dissolved oxygen:

7.6 - 8.7 mg O2/L

Nominal and measured concentrations:

A single nominal loading rate of 100 mg/L Water Accomodated Fraction was used. Measured concentrations were 0.0489 mg/L at 0 hours and less than the limit of quantification (LOQ) at 24 and 48 hours (in fresh and old media).

Details on test conditions:

TEST SYSTEM
- Test vessel: glass jar, covered to reduce evaporation
- Material, size, headspace, fill volume: 250 mL glass jar filled with approximately 200 mL of test preparation
- Aeration: no
- Renewal rate of test solution: every 24 hours
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water (ISO medium)
- pH: 7.8± 0.2
- Hardness: 250 mg/L as CaCO3
- Culture medium different from test medium: yes, Daphnia were cultured in Elendt M7 medium
- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS
- Photoperiod: 16 h of light and 8 h of darkness with 20 min dawn and dusk transition period
- Light intensity: 628 to 640 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): immobilization and adverse reaction to exposure were recorded at 24 and 48 hours after the start of exposure

TEST CONCENTRATIONS
- Range finding study: yes
- Test concentrations: 1.0, 10 and 100 mg/L (nominal loading rates WAF)
- Results used to determine the conditions for the definitive study: no immobilization was observed in the range finding test up to 100 mg/L loading rate WAF.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 other: mg/L loading rate WAF

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 other: mg/L loading rate WAF

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

There was no immobilization in 20 daphnids exposed to a 100 mg/L loading rate WAF for a period of 48 hours.

Results with reference substance (positive control):

- Results with reference substance valid? yes, the results from the positive control with potassium dichromate were within the normal range for the reference item.
- 48-h EC50: 0.71 mg/L (95% CL: 0.65-0.76)
- Other: NOEC = 0.32 mg/L, LOEC = 0.56 mg/L

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test item to the freshwater invertebrate Daphnia magna has been investigated and gave a 48-Hour EL50 value of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading rate was 100 mg/L loading rate WAF.

Executive summary:

A GLP-compliant study following OECD guideline 202 and EU Method C.2 was conducted using the test material Terpenes and terpenoids, turpentine oil, alpha-pinene fraction oligomers

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test item was prepared as a Water Accommodated Fraction (WAF). Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to a Water Accommodated Fraction (WAF) of the test item at single nominal loading rate of 100 mg/L for 48 hours at a temperature of approximately 22 °C under semi-static test conditions. The number of immobilized Daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours.

There were minor deviations from the guidelines but these are not considered to have affected the study results.

Exposure of Daphnia magna to the test item gave EL50 values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.

Description of key information

A GLP-compliant study following OECD guideline 202 and EU Method C.2 was conducted using the test material Terpenes and terpenoids, turpentine oil, alpha-pinene fraction oligomers

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test item was prepared as a Water Accommodated Fraction (WAF). Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to a Water Accommodated Fraction (WAF) of the test item at single nominal loading rate of 100 mg/L for 48 hours at a temperature of approximately 22 °C under semi-static test conditions. The number of immobilized Daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours.

There were minor deviations from the guidelines but these are not considered to have affected the study results.

Exposure of Daphnia magnato the test item gave EL50 values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L