Reaction mass of (4R)-isopropenyl-1-methylcyclohexene and (4S)-isopropenyl-1-methylcyclohexene and 1-isopropyl-4-methylcyclohexa-1,3-diene and 1-isopropyl-4-methylcyclohexa-1,4-diene and 4-isopropylidene-1-methylcyclohexene

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

calculation (if not (Q)SAR)

Remarks:

estimated by calculation

Adequacy of study:

key study

Study period:

2014-05-26 to 2014-05-28

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

accepted calculation method

Remarks:

Calculation method is used; calculation method applicable for the endpoint.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Calculation method

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

Calculation method

Principles of method if other than guideline:

The acute toxicity to fish was determined using a calculation method for the Mode of Action in question. This algorithm is based on a QSAR model which has been validated to be compliant with the OECD recommendations for QSAR modeling (OECD, 2004). Further to this the lethal loading rate of the WAF is determined by using a series of calculation steps using phase equilibrium thermodynamics and excluding the non-bioavailable fraction, this approach is based on validated data derived from 96-hour LL50 tests on fish, for which the concentrations of the test item had been determined by chemical analyses over the test period.

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

Not applicable

Vehicle:

no

Details on test solutions:

Not applicable

Test organisms (species):

other: Results from the following species were used in the regression: Danio rerio, Oncorhynchus mykiss, Lepomis macrochirus, Pimephales promelas, Oryzias latipes, Leuciscus idus

Details on test organisms:

Not applicable

Test type:

other: calculation method

Water media type:

not specified

Total exposure duration:

96 h

Remarks on exposure duration:

LL50 (lethal loading rate of WAF)

Post exposure observation period:

Not applicable

Hardness:

Hardness is not a necessary component of the WAF calculation

Test temperature:

The Temperature is not a necessary component of the WAF calculation but extremely low or high temperatures could influence the solubility of certain constituents. Therefore, the calculation method is considered acceptable to determine LL50s for fish between 12 and 28 °C.

pH:

The pH is not a necessary component of the WAF calculation

Dissolved oxygen:

The oxygen concentration is not a necessary component of the WAF calculation

Salinity:

Salinity is not a necessary component of the WAF calculation. However as the fish QSAR for the constituents calculation was based on data from freshwater studies, the resulting calculation is considered valid for freshwater fish

Nominal and measured concentrations:

The calculation determines measured concentrations

Details on test conditions:

Calculation method

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

6.7 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Not applicable

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not applicable

Sublethal observations / clinical signs:

Table 6.1.1/2: Expected concentrations at this 96 h-LL50

Constituents	concentration in the WAF (mg/L)
limonene	2.41
α-terpinene	1.20
γ-terpinene	0.70
terpinolene	0.84
isoterpinolene	0.30
para-cymene	0.14
camphene	0.31
α-phellandrene	0.04
α-pinene	0.03

Validity criteria fulfilled:

yes

Conclusions:

The 96 h-LL50 for dipentene multiconstituent is 6.7 mg/L.

Executive summary:

A calculation method prediction was performed to assess the acute toxicity of test item dipentene multiconstituent, a multiconstituent substance, to fish using the WAF method. This calculation method predicts the endpoint value which would be expected when testing the substance under experimental conditions in a laboratory following Guideline OECD 203 and EU method C.1 adapted for testing as a mixture using the WAF method. The criterion measured was the lethal loading rate of the mixture LL₅₀ (Median Lethal Loading), a statistically derived loading rate which is expected to cause mortality in 50 % of test animals within a period of 96 h.

The acute toxicity to fish was determined using a calculation method for the Mode of Action in question. This algorithm is based on a QSAR model which has been validated to be compliant with the OECD recommendations for QSAR modeling (OECD, 2004). Further to this the lethal loading rate of the WAF is determined by using a series of calculation steps using phase equilibrium thermodynamics and excluding the non-bioavailable fraction, this approach is based on validated data derived from 96 h LL50 tests on fish, for which the concentrations of the test item had been determined by chemical analyses over the test period.

The iSafeRat® mixture module first uses Phase Equilibrium Thermodynamics to determine the analytically measurable concentrations in aqueous solutions expected from known WAF loading rates of characterised mixtures for fish, daphnids or algae. These loading rates are then divided into two parts: the bioavailable and non-bioavailable fractions per constituent. Existing experimental ecotoxicity endpoint values or predictions of these (from the single constituent ecotoxicity modules) are then used to determine the mixture ecotoxicity based on the sum of the chemical activities of the remaining bioavailable fractions of the individual constituents. The resulting value is converted back to the equivalent loading rate providing a LL50 value.

The LC50s of each constituent are already known from literature or predicted using the iSafeRat QSAR model. QMRF and QPRF are available and attached in the endpoint study record. An additivity approach (based on Chemical Activity of each constituent) is used in order to calculate the Lethal Loading of the WAF.

Using this approach, the 96-h LL₅₀ for dipentene multiconstituent to fish is 6.7 mg/L. Based on the results of this study, dipentene multiconstituent would not be classified as acute 1 to aquatic organisms in accordance with CLP regulation (EC) No. 1272/2008.

This toxicity study is acceptable and can be used for that endpoint.

Description of key information

Using a calculation method to predict the acute toxicity of the registered substance, REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENE, to fish, the 96h-LL50 was found to be 6.7 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

6.7 mg/L

Additional information

A calculation method prediction was available and it has been performed to assess the acute toxicity of test item dipentene multiconstitent, a multiconstituent substance, to fish, using the WAF method. This calculation method predicts the endpoint value which would be expected when testing the substance under experimental conditions in a laboratory following Guideline OECD 203 and EU method C.1 adapted for testing as a mixture using the WAF method. The criterion measured was the lethal loading rate of the mixture LL50 (Median Lethal Loading), a statistically derived loading rate which is expected to cause mortality in 50% of test animals within a period of 96 h.

The algorithm used for the purpose of this study is based on a QSAR model which has been validated to be compliant with the OECD recommendations for QSAR modeling (OECD, 2004).

The 96h-LC50s of each constituent have been predicted using the iSafeRat QSAR model and the QMRF/QPRF have been attached to the dossier:

- p-cymene = 1.63 mg test item/L with 95 % - Confidence Limit of 1.21-2.20 mg test item/L.

- γ-terpinene = 1.09 mg test item/L with 95 % - Confidence Limit of 0.91 - 2.07 mg test item/L.

- α-phellandrene = 0.63 mg test item/L with 95 % - Confidence Limit of 0.43 - 0.92 mg test item/L.

- α-terpinene = 0.63 mg test item/L with 95 % - Confidence Limit of 0.43 - 0.92 mg test item/L.

- Isoterpinolene = 0.63 mg test item/L with 95 % - Confidence Limit of 0.43 - 0.92 mg test item/L.

- α-pinene = 0.44 mg test item/L with 95 % - Confidence Limit of 0.29 - 0.66 mg test item/L.

- Terpinolene = 1.09 mg test item/L with 95 % - Confidence Limit of 0.78 - 1.51 mg test item/L.

- Camphene = 0.44 mg test item/L with 95 % - Confidence Limit of 0.29 - 0.66 mg test item/L.

- Limonene = 1.09 mg test item/L with 95 % - Confidence Limit of 0.78 - 1.51 mg test item/L.

An additivity approach (based on Chemical Activity of each constituent) was used in order to calculate the Lethal Loading of the WAF. Using this approach, the 96-h LL50 for dipentene multiconstituent to fish is 6.7 mg/L. Based on the results of this study, dipentene multiconstituent would not be classified as acute 1 to aquatic organisms in accordance with CLP regulation (EC) No. 1272/2008. This toxicity study is acceptable and can be used for that endpoint.