2-(4-methylcyclohex-3-en-1-yl)propan-2-ol

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006-02-27 to 2006-03-06

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

GLP study conducted in accordance with the OECD guideline 203. Specificity of Method of analysis is unclear but the exposure medium was renewed every 24h (semi-static). Therefore the expoure of test organisms is expected to be close to the nominal.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 62 mg/L and 80 mg/L (abiotic test medium solutions)
- Sampling method: Analysis of test element was performed in the test solutions "test medium control" at t= 0 and t=24h (before the first renewal).
- Sample storage conditions before analysis: at 4 +-2°C

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: direct introduction of the test substance in each aquarium and addition of test medium in order to get a 2.5 L test solution.
- Eluate: none
- Differential loading: 28 mg/L, 36 mg/L, 47 mg/L, 62 mg/L, 80 mg/L
- Controls: test medium control
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Danio rerio
- Source: HB developpement élevage, La Fond Garel, 69490 St Forgeux.
- Length at study initiation (length definition, mean, range and SD): 25 to 30 mm
- Feeding during test: none

ACCLIMATION
- Acclimation period:12 days
- Acclimation conditions (same as test or not): same
- Type and amount of food: no data
- Feeding frequency: no data
- Health during acclimation (any mortality observed): no data

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

none

Hardness:

270 mg/L CaCO3

Test temperature:

23.2 to 25 °C

pH:

7.2 to 7.9

Dissolved oxygen:

78 to 98%

Salinity:

not relevant

Nominal and measured concentrations:

Nominal: 28 mg/L, 36 mg/L, 47 mg/L, 62 mg/L, 80 mg/L
Measured:
- for the 62 mg/l solution: at t= 0, 44.0% which corresponds to 62.5 mg/L when compared to the 100 mg/L solution reference
at t = 24h, 41.3 % which corresponds to 58.66 mg/L
- for the 80 mg/Lsolution: at t= 0, 54.6% which corresponds to 77.55 mg/L when compared to the 100 mg/L solution reference
at t = 24h, 50.4 % which corresponds to 71.59 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: all-glass vessel
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass, 15 cm*15cm*20 cm, contains 2.5 liters
- Aeration: no data
- Renewal rate of test solution (frequency/flow rate): 24h
- No. of organisms per vessel: 7
- Biomass loading rate: 0.9 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: theoritical composition 294.0 mg/L CaCl2, 2 H20, 123.3 mg/L MgSO4, 7 H2O, 63.0 mg/L NaHCO3, 5.5 mg/L KCl
- Total organic carbon: 0.02 mg/l

- Intervals of water quality measurement: 24 h

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 14h light /10 h dark
- Light intensity:no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
every 24h , the following parameters were checked: pH, dissolved oxygen, and temperature

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

>= 62 - <= 80 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

>= 62 - <= 80 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

>= 62 - <= 80 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

>= 62 - <= 80 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: fish at concentration 62 mg/L showed abnormal behaviour , they stayed on the side, at the bottom of the aquarium and showed a shortness of breath.
- Observations on body length and weight: no data
- Other biological observations: no data
- Mortality of control: none
- Other adverse effects control: none reported
- Abnormal responses: none reported
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none
- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality:
- LC50: 227 mg/L

Sublethal observations / clinical signs:

Table 6.1.1 -01: Cumulated dead fish as a function of exposure time and concentration

Nominal concentration of test substance	Fish number at the beginning of the test	Cumulated number of dead fish at each observation time
		6h	24h	48h	72h	96h
Control	7	0	0	0	0	0
28 mg/L	7	0	0	0	0	0
36 mg/L	7	0	0	0	0	0
47 mg/L	7	0	0	0	0	0
62 mg/L	7	0	0	0	0	0
80 mg/L	7	6	7	7	7	7

Table 6.1.1 -02: TOC analysis in the control medium solutions, 62 mg/L and 80 mg/L at the beginning (T =0) and at the fisrt renewal ( T= 24h)

Test solution	TOC at T = 0h	TOC at T= 24h
Control fish test medium	0.02	0.09
62 mg/L	44.0	41.3
80 mg/L	54.6	50.4

Validity criteria fulfilled:

yes

Conclusions:

The 96h LC50 of Terpineol multiconstituent on Danio rerio fish was measured to be between 62 and 80 mg/L.

Executive summary:

In a 96-h acute toxicity study, Danio rerio fish were exposed to Terpineol multiconstituent at concentrations of 0 (test medium control), 28, 36, 47, 62 and 80 mg/L} under semi-static conditions.  The 96-h LC₅₀was between 62 and 80 mg/L.  The NOEC value, based on mortality effects, was 62 mg/L.  Sublethal effects were observed at the concentration of 62 mg terpineol multiconstituent /L were observed: abnormal behaviour , the fish stayed on the side, at the bottom of the aquarium and showed a shortness of breath.

 

This toxicity study is classified as acceptable and satisfies the guideline requirements for fish short term toxicity study.

 

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

calculation (if not (Q)SAR)

Remarks:

Migrated phrase: estimated by calculation

Adequacy of study:

key study

Study period:

2016-03-07 to 2016-03-08

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

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

Principles of method if other than guideline:

The acute toxicity to fish was determined using a validated QSAR for the Mode of Action in question. The first step of the iSafeRat mixture toxicity calculation employs phase equilibrium thermodynamics in order to determine the concentrations of each constituent within the WAF. This fraction equates to the analyzable fraction of a WAF study.
Within the WAF, the constituents also partition between themselves further reducing the bioavailable fraction and thus the toxicity of the mixture compared to the individual constituents. In the calculation the second step is to remove this non-bioavailable fraction.
The final step is to determine the truly bioavailable fraction of the WAF per constituent. The LC50s of each constituent are already known from literature or calculated using the iSafeRat QSAR model. Each value and calculation has been included as a supporting study in the IUCLID. An additivity approach (based on Chemical Activity of each constituent) is used in order to calculate the Lethal Loading of the WAF.
The method has been validated using data derived from 96-hour LC50 tests on fish, for which the concentrations of the test item had been determined by chemical analyses over the test period. 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.

GLP compliance:

no

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
not applicable

Analytical monitoring:

not required

Details on sampling:

not applicable

Vehicle:

no

Details on test solutions:

not applicable

Test organisms (species):

other: fish spp.

Details on test organisms:

not applicable

Test type:

other: calculation method

Water media type:

freshwater

Limit test:

no

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.:

ca. 12 mg/L

Nominal / measured:

estimated

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: typical composition (please refer to confidential section)

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 6.1.1/1: Short-term toxicity to fish values for components.

constituents	96h-LC50 (mg/L)
α-terpineol	69
γ-terpineol	69
β-terpineol	57
terpinolene	0.55
3-terpinen-1-ol	69
borneol	23
fenchol	23
terpinen-1-ol 4	69
limonene	0.46

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

constituents	concentration in the WAF (mg/L)
α-terpineol	7.6
γ-terpineol	1.2
β-terpineol	0.63
terpinolene	1.3
3-terpinen-1-ol	0.43
borneol	0.49
fenchol	0.29
terpinen-1-ol 4	0.20
limonene	0.07

Validity criteria fulfilled:

yes

Conclusions:

The 96h-LL50 for Terpineol multiconstituent is estimated at 12 mg test item/L.

Executive summary:

The registered substance is a multiconstituent substance with a well-defined composition for which the relative percentage and the reported variation of each constituent is known.

Its acute toxicity to fish property has been investigated using an in-house calculation method that replaces an OECD 203 study and guideline for Testing of Chemicals No. 203 (i.e. WAF conditions). The worst case composition has been investigated.

The first step of the iSafeRat mixture toxicity calculation employs phase equilibrium thermodynamics in order to determine the concentrations of each constituent within the WAF. This fraction equates to the analyzable fraction of a WAF study.

Within the WAF, the constituents also partition between themselves further reducing the bioavailable fraction and thus the toxicity of the mixture compared to the individual constituents. In the calculation the second step is to remove this non-bioavailable fraction.

The final step is to determine the truly bioavailable fraction of the WAF per constituent. The LC50s of each constituent are already known from literature or calculated using the iSafeRat QSAR model. Each value and calculation has been included as a supporting study in IUCLID. An additivity approach (based on Chemical Activity of each constituent) is used in order to calculate the Lethal Loading of the WAF.

The 96-h LL50 was 12 mg test material/L for the assessed composition. 96h-LL50 were based on mortality. 

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

 

Description of key information

The substance exhibits a LC50 for freshwater fish of 12 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

12 mg/L

Additional information

The registered substance is a multiconstituent substance with a well-defined composition for which the relative percentage of each constituent/impurity is known. The ecotoxicological properties of the registered substance may be derived from the aquatic toxicity values of the individual components using an additivity calculation approach.

It has been observed that CLP additivity calculations for mode of action 1 (MOA 1) compounds are unreasonably conservative when compared to classic experimental studies using Water Accommodated Fractions (WAF), and this has been proved in a number of cases with substances of natural origin. Indeed, substance of natural origin may be composed of hydrophilic alcohol molecules and hydrophobic hydrocarbon terpene molecules. Therefore, when a WAF is performed most of substances fully dissolved in the aqueous phase are the hydrophilic fraction while the hydrophobic fraction (the more toxic elements for MOA 1 substances) may be below their water solubility value. The first step of the iSafeRat mixture toxicity calculation employs phase equilibrium thermodynamics in order to determine the concentrations of each constituent within the WAF. This fraction equates to the analysable fraction of a WAF study. On the other hand, the CLP additivity approach is calculated on the basis that all the substances are at their maximum solubility. Within the WAF, the constituents also partition between themselves further reducing the bioavailable fraction and thus the toxicity of the mixture compared to the individual constituents. In the calculation the second step is to remove this non-bioavailable fraction. These two reasons explain why ecotoxicity values from WAF studies are always higher for non-polar narcotic mixtures than the calculated values from CLP additivity calculation. The final step is to determine the truly bioavailable fraction of the WAF per constituent. The LC₅₀s of each constituent are already known from literature or calculated using the iSafeRat QSAR model. Each value and calculation has been included as a supporting study in the IUCLID. An additivity approach (based on Chemical Activity of each constituent) is used in order to calculate the Lethal Loading of the WAF.

A predicted acute toxicity value to fish is available. The estimated LC₅₀ for freshwater fish is 12 mg/L. This prediction is obtained with QSAR for the Mode of Action 1, based on validated data derived from standard toxicity test, for which the concentrations of the test item had been determined by chemical analyses over the test period. The prediction is sufficiently robust, and is appropriate for the purposes of chemical safety assessment required for REACH.

Besides, one reliable experimental study is available for the registered substance. The toxicity to freshwater fish was studied according to OECD TG No. 203 and GLP. Fish (Danio rerio) were exposed to substance at nominal test concentrations of 0 (test medium control), 28, 36, 47, 62 and 40 mg/L over a test period of 96 hours (semi-static test, daily renewal). Concentrations were verified by TOC determinations at relevant concentrations (62 and 80 mg/L). The endpoint was based on the nominal concentrations. The 96-h LC₅₀ was approximately between 62 and 80 mg/L. Sublethal effects were observed at the concentration of 62 mg of terpineol multiconstituent /L: abnormal behaviour, the fish stayed on the side, at the bottom of the aquarium and showed a shortness of breath.