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Ecotoxicological information

Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 Feb 2014 - 26 Feb 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
Feeding was discontinued approximately 23 hours prior testing. This was not considered to have had an effect on the outcome or integrity of the study.
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
yes
Remarks:
Feeding was discontinued approximately 23 hours prior testing. This was not considered to have had an effect on the outcome or integrity of the study.
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. certificate)
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:
- Concentrations: Samples were taken from the control and the 100 mg/L loading rate WAF test vessel at 0 and 0 and 72 hours (fresh media) and 24 and 96 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: Water Accommodated Fraction (WAF)
A nominal amount of test item (2200 mg) was weighted onto a glass slide and suspended in the water column of 22 liters 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 for 23 hours. Then the stirring was stopped and the mixture was allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Necsofilm 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 microdispersions or undissolved test item to be present. The aqueous phase of the WAF was removed by mid-depth siphoning (the first 75-100 mL discarded) to give the 100 mg/L loading rate WAF.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: rainbow trout
- Source: Brow Well Fisheries Limited, Hebden near Skipton, Yorkshire, UK
- Age at study initiation (mean and range, SD): juvenile fish
- Length at study initiation (length definition, mean, range and SD): not reported, 4.4 cm (SD = 0.2) at the end of the test
- Weight at study initiation (mean and range, SD): not reported, 1.01 g (SD = 0.15) at the end of the test
- Feeding during test: no


ACCLIMATION
- Acclimation period: from 05 February 2014 to 17 February 2014
- Acclimation conditions (same as test or not): the same as during the test
- Type and amount of food: commercial trout pellets
- Feeding frequency: not reported
- Health during acclimation (any mortality observed): no mortality in the 7 days prior to the start of the test
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
approximately 140 mg/L as CaCO3
Test temperature:
14 - 15°C
pH:
7.6 - 8.1
Dissolved oxygen:
9.5 - 10.5 mg O2/L
Salinity:
N/A
Nominal and measured concentrations:
A single nominal loading rate of 100 mg/L Water Accommodated Fraction was used. Measured concentrations were 0.0071 mg/L at 0 hours and less than the limit of quantification (LOQ) at 72 hours in fresh media. Measured concentrations were 0.055 mg/L and less than LOQ at 24 and 96 hours in old media.
Details on test conditions:
TEST SYSTEM
- Test vessel: 20 L glass exposure vessels
- Aeration: Yes, the test vessels were aerated via narrow bore glass tubes.
- Renewal rate of test solution (frequency/flow rate):daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.35 g bodyweight/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ?Laboratory tap water, dechlorinated by passage through an activated carbon filter and partly softened (approx. 140 mg/L as CaCO3), then passed through a series of computer controlled plate heat exchangers to achieve the required temperature.
- Culture medium different from test medium: no
- Intervals of water quality measurement: 24 hours

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition period
- Light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : any mortalities and sub-lethal effects were recorded at 3, 6, 24, 48, 72 and 96 hours after the start of exposure.

TEST CONCENTRATIONS
- Test concentrations: the test was conducted at a single loading rate of 100 mg/L loading rate WAF.
Key result
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 100 other: mg/L loading rate WAF
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
100 other: mg/L loading rate WAF
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
There were no mortalities in 7 fish exposed to a 100 mg/L loading rate WAF for a period of 96 hours.
Results with reference substance (positive control):
Yes

Specificity:

The biological control samples and an analised blank showed no significant interfering response at the retention time of the test item. The standard solutions contained a peak specific for the test item whose area changed accordingly with known concentration, hence the specificity of the method by retention time was confirmed.

Linearity

The data was found to have a quadratic correlation within the claibration range of 0 to 22 mg/L. The R2 fit of the calibration curve to the data was 0.9993 and was considered to be acceptable.

Accuracy and Precision:

A set of recovery samples accurately fortified at a relevant concentration of the test item was prepared five-fold and analized.

LOQ:

The limit of quatification (LOQ) was determined by calculating the sample concentration that gave a peak equivalent to ten times the baseline noise. Using this method the LOQ was determined to be 0.0027 mg/L.

Validity criteria fulfilled:
yes
Remarks:
None of the control fish died or showed signs of stress during the test. Oxygen concentration at the end of the test was ≥60% of ASV in the control and test vessles.
Conclusions:
The acute toxicity of the test item to the freshwater fish rainbow trout (Oncorynchus mykiss) has been investigated using the threshold approach and gave a 96-hour LL50 value of greater than 100 mg/L loading rate WAF. The NOEL rate was 100 mg/L loading rate WAF.
Executive summary:

A GLP-compliant study following OECD guideline 203 and EU Method C.1 was conducted using the substance 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). Seven fish were exposed to a Water Accommodated Fraction (WAF) of the test item at single nominal loading rate of 100 mg/L for 96 hours at a temperature of approximately 14 °C under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure were recorded after 3 and 6 hours and then daily throughout the test until termination after 96 hours.

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

Exposure of rainbow trout to the test item gave LL50 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 203 and EU Method C.1 was conducted using the substance 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). Seven fish were exposed to a Water Accommodated Fraction (WAF) of the test item at single nominal loading rate of 100 mg/L for 96 hours at a temperature of approximately 14 °C under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure were recorded after 3 and 6 hours and then daily throughout the test until termination after 96 hours.

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

Exposure of rainbow trout to the test item gave LL50 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

LC50 for freshwater fish:
100 mg/L

Additional information