Reaction mass of 7,7-dimethyl-2-methylidenebicyclo[2.2.1]heptane and (1R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane and (1S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane and (1S)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene

Toxicity to terrestrial arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to terrestrial arthropods: long-term

Data waiving:

exposure considerations

Justification for data waiving:

the study does not need to be conducted because direct and indirect exposure of the soil compartment is unlikely

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
In accordance with column 2 of REACH Annex IX, the study does not need to be conducted since direct and indirect exposure of the soil compartments is unlikely.

Reference 2

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

A reference substance (toxic standard) was not included in the experimental design. No data on GLP.

Qualifier:

no guideline followed

Principles of method if other than guideline:

The toxicity of alpha-pinene to Dendroctonus rufipennis (spruce beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays.

GLP compliance:

not specified

Application method:

spray

Analytical monitoring:

not specified

Vehicle:

no

Details on preparation and application of test substrate:

-Method of application: Adult spruce beetles were exposed to alpha pinene concentrations in the air
- Controls: Control group of water alone.

Test organisms (species):

other: Dendroctonus rufipennis (spruce beetle )

Animal group:

Coleoptera (soil-dwelling rove beetle)

Details on test organisms:

TEST ORGANISM
- Common name: Bettles
- Source: Beetles used in the bioassays were collected near Fairbanks, AK, from overwintering sites at the base of trees in early May, 2 weeks before the period of flight.
- Age at test initiation (mean and range, SD): Adults

Study type:

laboratory study

Limit test:

no

Total exposure duration:

24 h

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 250-mL jar with screened tops.
- No. of organisms per container (treatment): 1
- No. of replicates per treatment group: 5
- No. of replicates per control: 1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mortality. Beetles were considered dead if they did not move when probed with a forceps.

Nominal and measured concentrations:

Nominal concentrations: 20, 40, 60 and 80 ppm.

Reference substance (positive control):

no

Key result

Duration:

5.9 h

Dose descriptor:

other: LT50

Effect conc.:

80 ppm

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: (95 % CL: 4.5 - 18.8 h)

Details on results:

LT50 (Time to 50% population mortality): 5.9 h at a concentration of 80 ppm.
Mortality of 100 % of beetles at 80 ppm after 24 hours of exposure.

Reported statistics and error estimates:

Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. Analysis of variance (ANOVA) (SAS Institute 1982) was used for all statistical tests.

The lethal time (LT50, time to 50% population mortality) for Dendroctonus rufipennis (spruce beetle ) was 5.9 hours. Mortality of 100 % of beetles at 80 ppm was observed after 24 hours of exposure.

Validity criteria fulfilled:

no

Conclusions:

The lethal time (LT50, time to 50% population mortality) for Dendroctonus rufipennis (spruce beetle ) was 5.9 hours at 80 ppm, and a mortality of 100 % of beetles was observed after 24 hours of exposure.

Executive summary:

The toxicity of alpha pinene to Dendroctonus rufipennis (spruce beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus rufipennis (spruce beetle ) was 5.9 hours. Mortality of 100 % of bettles at 80 ppm was observed after 24 hours of exposure.

Reference 3

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

A reference substance (toxic standard) was not included in the experimental design. No data on GLP.

Qualifier:

no guideline followed

Principles of method if other than guideline:

The toxicity of Camphene to Dendroctonus rufipennis (spruce beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays.

GLP compliance:

not specified

Application method:

spray

Analytical monitoring:

not specified

Vehicle:

no

Details on preparation and application of test substrate:

-Method of application: Adult spruce beetles were exposed to camphene concentrations in the air
- Controls: Control group of water alone.

Test organisms (species):

other: Dendroctonus rufipennis (spruce beetle )

Animal group:

Coleoptera (soil-dwelling rove beetle)

Details on test organisms:

TEST ORGANISM
- Common name: Bettles
- Source: Beetles used in the bioassays were collected near Fairbanks, AK, from overwintering sites at the base of trees in early May, 2 weeks before the period of flight.
- Age at test initiation (mean and range, SD): Adults

Study type:

laboratory study

Limit test:

no

Total exposure duration:

24 h

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 250-mL jar with screened tops.
- No. of organisms per container (treatment): 1
- No. of replicates per treatment group: 5
- No. of replicates per control: 1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mortality. Beetles were considered dead if they did not move when probed with a forceps.

Nominal and measured concentrations:

Nominal concentrations: 20, 40, 60 and 80 ppm.

Reference substance (positive control):

no

Key result

Duration:

12.9 h

Dose descriptor:

other: LT50

Effect conc.:

80 ppm

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: (95 % CL: 9.9 - 39.4 h)

Details on results:

LT50 (Time to 50% population mortality): 12.9 h at a concentration of 80 ppm.
Mortality of 83 % of beetles at 80 ppm after 24 hours of exposure.

Reported statistics and error estimates:

Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. Analysis of variance (ANOVA) (SAS Institute 1982) was used for all statistical tests.

The lethal time (LT50, time to 50% population mortality) for Dendroctonus rufipennis (spruce beetle ) was 12.9 hours. Mortality of 83 % of beetles at 80 ppm was observed after 24 hours of exposure.

Validity criteria fulfilled:

no

Conclusions:

The lethal time (LT50, time to 50% population mortality) for Dendroctonus rufipennis (spruce beetle ) was 12.9 hours at 80 ppm, and a mortality of 83 % of beetles was observed after 24 hours of exposure.

Executive summary:

The toxicity of Camphene to Dendroctonus rufipennis (spruce beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus rufipennis (spruce beetle ) was 12.9 hours. Mortality of 83 % of bettles at 80 ppm was observed after 24 hours of exposure.

Reference 4

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

A reference substance (toxic standard) was not included in the experimental design. No data on GLP.

Qualifier:

no guideline followed

Principles of method if other than guideline:

The toxicity of Camphene to Dendroctonus simplex (eastern larch beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays.

GLP compliance:

not specified

Application method:

spray

Analytical monitoring:

not specified

Vehicle:

no

Details on preparation and application of test substrate:

Adult spruce beetles were exposed to camphene concentrations in the air
- Controls: Control group of water alone.

Test organisms (species):

other: Dendroctonus simplex (eastern larch beetle )

Animal group:

Coleoptera (soil-dwelling rove beetle)

Details on test organisms:

TEST ORGANISM
- Common name: Bettles
- Source: Beetles used in the bioassays were collected near Fairbanks, AK, from overwintering sites at the base of trees in early May, 2 weeks before the period of flight.
- Age at test initiation (mean and range, SD): Adults

Study type:

laboratory study

Limit test:

no

Total exposure duration:

24 h

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 250-mL jar with screened tops.
- No. of organisms per container (treatment): 1
- No. of replicates per treatment group: 5
- No. of replicates per control: 1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mortality. Beetles were considered dead if they did not move when probed with a forceps.

Nominal and measured concentrations:

Nominal concentrations: 20, 40, 60 and 80 ppm.

Reference substance (positive control):

no

Key result

Duration:

16.3 h

Dose descriptor:

other: LT50

Effect conc.:

80 ppm

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: (95 % CL: 11.2 - 23.2 h)

Details on results:

LT50 (Time to 50% population mortality): 16.3 h at a concentration of 80 ppm.
Mortality of 77 % of beetles at 80 ppm after 24 hours of exposure.

Reported statistics and error estimates:

Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. Analysis of variance (ANOVA) (SAS Institute 1982) was used for all statistical tests.

The lethal time (LT50, time to 50% population mortality) for Dendroctonus simplex (eastern larch beetle ) was 16.3 hours. Mortality of 77 % of beetles at 80 ppm was observed after 24 hours of exposure.

Validity criteria fulfilled:

no

Conclusions:

The lethal time (LT50, time to 50% population mortality) for Dendroctonus simplex (eastern larch beetle ) was 16.3 hours at 80 ppm and a mortality of 77 % of beetles was observed after 24 hours of exposure.

Executive summary:

The toxicity of Camphene to Dendroctonus simplex (eastern larch beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus simplex (eastern larch beetle ) was 16.3 hours. Mortality of 77 % of bettles at 80 ppm was observed after 24 hours of exposure.

Reference 5

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

A reference substance (toxic standard) was not included in the experimental design. No data on GLP.

Qualifier:

no guideline followed

Principles of method if other than guideline:

The toxicity of alpha pinene to Dendroctonus simplex (eastern larch beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays.

GLP compliance:

not specified

Application method:

spray

Analytical monitoring:

not specified

Vehicle:

no

Details on preparation and application of test substrate:

Adult spruce beetles were exposed to alpha pinene concentrations in the air
- Controls: Control group of water alone.

Test organisms (species):

other: Dendroctonus simplex (eastern larch beetle )

Animal group:

Coleoptera (soil-dwelling rove beetle)

Details on test organisms:

TEST ORGANISM
- Common name: Bettles
- Source: Beetles used in the bioassays were collected near Fairbanks, AK, from overwintering sites at the base of trees in early May, 2 weeks before the period of flight.
- Age at test initiation (mean and range, SD): Adults

Study type:

laboratory study

Limit test:

no

Total exposure duration:

24 h

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 250-mL jar with screened tops.
- No. of organisms per container (treatment): 1
- No. of replicates per treatment group: 5
- No. of replicates per control: 1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mortality. Beetles were considered dead if they did not move when probed with a forceps.

Nominal and measured concentrations:

Nominal concentrations: 20, 40, 60 and 80 ppm.

Reference substance (positive control):

no

Key result

Duration:

3.6 h

Dose descriptor:

other: LT50

Effect conc.:

80 ppm

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: (95 % CL: 2.1 - 6.3 h)

Details on results:

LT50 (Time to 50% population mortality): 3.6 h at a concentration of 80 ppm.
Mortality of 100 % of beetles at 80 ppm after 24 hours of exposure.

Reported statistics and error estimates:

Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. Analysis of variance (ANOVA) (SAS Institute 1982) was used for all statistical tests.

The lethal time (LT50, time to 50% population mortality) for Dendroctonus simplex (eastern larch beetle ) was 3.6 hours. Mortality of 100 % of beetles at 80 ppm was observed after 24 hours of exposure.

Validity criteria fulfilled:

no

Conclusions:

The lethal time (LT50, time to 50% population mortality) for Dendroctonus simplex (eastern larch beetle ) was 3.6 hours at 80 ppm, and a mortality of 100 % of beetles was observed after 24 hours of exposure.

Executive summary:

The toxicity of alpha pinene to Dendroctonus simplex (eastern larch beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus simplex (eastern larch beetle ) was 3.6 hours. Mortality of 100 % of bettles at 80 ppm was observed after 24 hours of exposure.

Reference 6

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

According to a test method suggested by the World Health Organization (WHO 2005) with modifications.

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: A larval mortality bioassay was carried out according to the test method of larval susceptibility as suggested by the World Health Organization (WHO 2005) with modifications.
- Short description of test conditions: see below
- Parameters analysed / observed: LC50 value

GLP compliance:

not specified

Application method:

contact

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of test material application: mosquito larvae were placed in 2 % (v/v) aqueous solution of dimethyl sulfoxide (DMSO) (98 ml of tap water plus 2 ml of DMSO), followed by addition of the tested material solution and gentle shaking to ensure homogeneous test solution.
- Controls: 2 % (v/v) aqueous solution of DMSO (98 ml of tap water plus 2 ml of DMSO)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dimethyl sulfoxide (DMSO)
- Stock solution (tested material solution): 10 % (w/v) in dimethyl sulfoxide (DMSO)

Test organisms (species):

other: Aedes albopictus

Animal group:

Diptera (mosquitoe)

Details on test organisms:

TEST ORGANISM
- Common name: Asian tiger mosquito
- Source: laboratory of Benaki Phytopathological Institute, Kifissia, Greece. Colony conditions maintained: 25±2 °C, 80 % relative humidity, and photoperiod of 16:8-h light/dark (L/D).
- Stage at test initiation: late third- to early fourth-instar mosquito larvae
- Kept according to standard practices: yes

ACCLIMATION
- Acclimation conditions: Larvae were reared in tap water-filled cylindrical enamel pans with diameter of 35 and 10 cm deep covered by fine muslin
- Feeding: Approximately 400 larvae were fed ad libitum with powdered fish food (JBL Novo Tom 10 % Artemia) in each pan until the adults emerged.

Study type:

laboratory study

Limit test:

yes

Total exposure duration:

24 h

Details on test conditions:

TEST SYSTEM
- Test container (material, size): vial
- No. of organisms per container (treatment): 20
- No. of replicates per treatment group: 4
- No. of replicates per control: 1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): total mortality of mosquito larvae (milligram per liter of concentration in water)

Reference substance (positive control):

no

Key result

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

72.3 ppm

Nominal / measured:

estimated

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: (95 % CL: 67.79 - 76.49 ppm)

Reported statistics and error estimates:

LC50 was determined by probit analysis in which probit transformed mortality was regressed against log10-transformed dose (Finney 1971). Analysis were conducted using the statistical package SPSS 14.0 (SPSS Inc., Chicago, IL, 2004).

Validity criteria fulfilled:

not applicable

Conclusions:

The 24 h larvicidal LC50 for the test item on Aedes albopictus (Asian tiger Mosquito) was estimated to be 72.3 mg/L (95 % CL: 67.79-76.49)

Executive summary:

A larvicidal bioassay on Aedes albopictus (Asian tiger Mosquito) was conducted for some citrus essential oils and in particular for one of their components L-alpha pinene in accordance to the test method of larval susceptibility as suggested by the World Health Organization (WHO 2005). Twenty late third-to early fourth-instar mosquito larvae were placed in 2 % v/v aqueous solution of DMSO (98 ml of tap water plus 2 ml of DMSO), followed by addition of the tested material solution (10% w/v in DMSO). Four replicates and one negative control (2% v/v in DMSO) were tested. After 24 h exposure the mortality was recorded and the LC50 of the test item, determined by probit analysis, was estimated to be 72.3 mg/L (95 % CL: 67.79-76.49).

Reference 7

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Assessment of repellent activity of the test compound based on the human landing counts (Coleman et al. 1993; Govere and Durrheim 2006).
- Short description of test conditions: see below
- Parameters analysed / observed: human landing counts of test organisms

GLP compliance:

not specified

Application method:

contact

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of test material application: The test material was applied on paper (Whatman chromatography paper) of 24 cm2 total area placed around the opening measuring of 5×5 cm made in a plastic glove.
- Body part: human hand
- Volume of test solution applied: 50 μl (“low”, ≈0.2 μlcm−2 of testing material) and 100 μl (“high”, ≈0.4 μlcm−2 of testing material) of 100 μgμl−1 stock solution
- Controls:
Negative control: test system without test material
Positive control: N,N-diethyl-3-methylbenzamide (DEET). Source: Bayer CropScience. Different doses (from 0.05 to 1 μlcm−2) for DEET were applied and found that the lowest dose, where zero landings were counted, was ≈0.2 μlcm−2.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dimethyl sulfoxide (DMSO)
- Stock solution (tested material solution): 10 % (w/v) in dimethyl sulfoxide (DMSO)
- Evaporation of vehicle before use: yes, during five minutes after application.

Test organisms (species):

other: Aedes albopictus

Animal group:

Diptera (mosquitoe)

Details on test organisms:

TEST ORGANISM
- Common name: Asian tiger mosquito
- Source: laboratory of Benaki Phytopathological Institute, Kifissia, Greece. Colony conditions maintained: 25±2 °C, 80 % relative humidity, and photoperiod of 16:8-h light/dark (L/D).
- Stage at test initiation: adult 5- to 10-day old
- Kept according to standard practices: yes

ACCLIMATION
- Acclimation conditions: Adult mosquitoes were kept in wooden framed cage (33×33×33 cm) with a 32×32 mesh, with easy access to 10 % sucrose solution on a cotton wick
- Feeding: Females were blood fed from primary author’s forearmonce a fortnight.

Study type:

laboratory study

Limit test:

no

Total exposure duration:

5 min

Test temperature:

25±2 °C

Humidity:

70–80 % relative humidity

Details on test conditions:

TEST SYSTEM
- Test container / cage (material, size): The study was conducted into a cage (33×33×33 cm) with a 32×32 mesh and with a 20-cm diameter circular opening fitted with cloth sleeve.
- No. of organisms per container (treatment): 100 adult mosquitoes (sex ratio, 1:1), starved for 12 h at 25±2 °C, and 70–80 % relative humidity.
- No. of replicates per treatment group: 8 (and 4 human volunteers)
- No. of replicates per control / vehicle control: 8 (and 4 human volunteers)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): human landing counts of test organisms

VEHICLE CONTROL PERFORMED: no

TEST CONCENTRATIONS
- Test concentrations: 50 μl (“low”, ≈0.2 μlcm−2 of testing material) and 100 μl (“high”, ≈0.4 μlcm−2 of testing material) of 100 μgμl−1 stock solution.

Nominal and measured concentrations:

Nominal: 0.2 μlcm−2 and 0.4 μlcm−2

Reference substance (positive control):

yes

Remarks:

N,N-diethyl-3-methylbenzamide (DEET)

Key result

Duration:

5 min

Dose descriptor:

other: NOEL

Effect conc.:

0.2 other: μlcm−2

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Average number of landings did not differ significantly (P≤0.05) from the negative

Key result

Duration:

5 min

Dose descriptor:

other: LOEL

Effect conc.:

0.4 other: μlcm−2

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Average number of landings did differ significantly (P≤0.05) from the negative

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: DEET offered full protection (zero landings) for both doses.

Reported statistics and error estimates:

Repellency (mosquito landings) was analyzed using Kruskal–Wallis test. When significant differences were detected, Mann–Whitney U tests were carried out for pair-wise comparison. Bonferroni correction was applied to correct for 66 pair-wise comparisons leading to an adjusted a=0.0006.

Validity criteria fulfilled:

not applicable

Conclusions:

After a study on repellent activity against Aedes albopictus (Asian tiger mosquito), the NOEL of the test item was determined to be 0.2 μlcm−2.

Executive summary:

A repellent activity assay against Aedes albopictus (Asian tiger mosquito) was conducted for some citrus essential oils and in particular for one of their components L-alpha pinene, based on the human landing counts (Coleman et al. 1993; Govere and Durrheim 2006). The test material was applied on filter paper of 24 cm2 total area placed around the opening measuring of 5×5 cm made in a plastic glove, which was inserted for 5 min into the cage containing 100 adult mosquitoes. Two doses of test item (0.2 μlcm−2 and 0.4 μlcm−2), negative control and positive control (N,N-diethyl-3-methylbenzamide (DEET)) were tested and repeated eight times. Based on average number of landings compared with the controls, the NOEL of the test item was determined to be 0.2 μlcm−2.

Reference 8

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
The analogue substance L-alpha pinene which shares the same functional groups with the main components of the multi-constituent substance reaction mass of fenchene and laevo camphene and dextro camphene and laevo alpha pinene also has comparable values for the relevant molecular properties.
See attached the reporting format.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

72.3 ppm

Nominal / measured:

estimated

Conc. based on:

other: Read-across from an analogue

Basis for effect:

mortality

Remarks on result:

other: read-across from an analogue for which LC50 (24 h) = 72.3 ppm (95 % CL: 67.79 - 76.49 ppm)

Validity criteria fulfilled:

not applicable

Conclusions:

Based on the read-across approach from the analogue L-alpha pinene, the 24 h larvicidal LC50 for the reaction mass on Aedes albopictus (Asian tiger Mosquito) was estimated to be 72.3 mg/L (95 % CL: 67.79-76.49)

Executive summary:

A larvicidal bioassay on Aedes albopictus (Asian tiger Mosquito) was conducted for some citrus essential oils and in particular for one of their components L-alpha pinene in accordance to the test method of larval susceptibility as suggested by the World Health Organization (WHO 2005). Twenty late third-to early fourth-instar mosquito larvae were placed in 2 % v/v aqueous solution of DMSO (98 ml of tap water plus 2 ml of DMSO), followed by addition of the tested material solution (10% w/v in DMSO). Four replicates and one negative control (2% v/v in DMSO) were tested. After 24 h exposure the mortality was recorded and the LC50 of the test item, determined by probit analysis, was estimated to be 72.3 mg/L (95 % CL: 67.79-76.49). Based on these results, the read-across approach was applied and the 24 h larvicidal LC50 for the reaction mass was estimated to be 72.3 mg/L (95 % CL: 67.79-76.49).

Reference 9

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
The analogue substance L-alpha pinene which shares the same functional groups with the main components of the multi-constituent substance reaction mass of fenchene and laevo camphene and dextro camphene and laevo alpha pinene also has comparable values for the relevant molecular properties.
See attached the reporting format.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

5 min

Dose descriptor:

other: NOEL

Effect conc.:

0.2 other: μlcm−2

Nominal / measured:

estimated

Conc. based on:

other: Read-across from an analogue

Basis for effect:

other: Average number of landings did not differ significantly (P≤0.05) from the negative

Remarks on result:

other: read-across from an analogue for which NOEL = 0.2 μlcm−2

Key result

Duration:

5 min

Dose descriptor:

other: LOEL

Effect conc.:

0.4 other: μlcm−2

Nominal / measured:

estimated

Conc. based on:

other: Read-across from an analogue

Basis for effect:

other: Average number of landings did differ significantly (P≤ 0.05) from the negative

Remarks on result:

other: read-across from an analogue for which LOEL = 0.4 μlcm−2

Validity criteria fulfilled:

not applicable

Conclusions:

Based on the read-across approach from the analogue L-alpha pinene, the NOEL of the reaction mass was determined to be 0.2 μlcm−2 after a study on repellent activity against Aedes albopictus (Asian tiger mosquito)

Executive summary:

A repellent activity assay against Aedes albopictus (Asian tiger mosquito) was conducted for some citrus essential oils and in particular for one of their components L-alpha pinene, based on the human landing counts (Coleman et al. 1993; Govere and Durrheim 2006). The test material was applied on filter paper of 24 cm2 total area placed around the opening measuring of 5×5 cm made in a plastic glove, which was inserted for 5 min into the cage containing 100 adult mosquitoes. Two doses of test item (0.2 μlcm−2 and 0.4 μlcm−2), negative control and positive control (N,N-diethyl-3-methylbenzamide (DEET)) were tested and repeated eight times. Based on average number of landings compared with the controls, the NOEL of the test item was determined to be 0.2 μlcm−2. Based on these results, the read-across approach was applied and the NOEL of the reaction mass was determined to be 0.2 μlcm−2.

Reference 10

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
The analogue substance alpha pinene which shares the same functional groups with the main components of the multi-constituent substance reaction mass of fenchene and laevo camphene and dextro camphene and laevo alpha pinene also has comparable values for the relevant molecular properties.
See attached the reporting format.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

5.9 h

Dose descriptor:

other: LT50

Effect conc.:

80 ppm

Nominal / measured:

estimated

Conc. based on:

other: Read-across from an analogue

Basis for effect:

mortality

Remarks on result:

other: read-across from an analogue for which LT50 = 5.9 h (95 % CL: 4.5 - 18.8 h)

Details on results:

Based on read across from the analogue alpha pinene, the following results were estimated for the reaction mass:
LT50 (Time to 50% population mortality): 5.9 h at a concentration of 80 ppm.
Mortality of 100 % of beetles at 80 ppm after 24 hours of exposure.

Validity criteria fulfilled:

not applicable

Conclusions:

Based on the read-across approach from the analogue alpha pinene, the lethal time LT50 of reaction mass for Dendroctonus rufipennis (spruce beetle ) was 5.9 hours at 80 ppm, and a mortality of 100 % after 24 hours of exposure was estimated.

Executive summary:

The toxicity of alpha pinene to Dendroctonus rufipennis (spruce beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus rufipennis (spruce beetle ) was 5.9 hours. Mortality of 100 % of bettles at 80 ppm was observed after 24 hours of exposure. Based on these results, the read-across approach was applied and the lethal time LT50 of reaction mass for Dendroctonus rufipennis (spruce beetle ) was determined to be 5.9 hours at 80 ppm, and a mortality of 100 % after 24 hours of exposure was estimated.

Reference 11

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
The analogue substance camphene which shares the same functional groups with the main components of the multi-constituent substance reaction mass of fenchene and laevo camphene and dextro camphene and laevo alpha pinene also has comparable values for the relevant molecular properties.
See attached the reporting format.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

12.9 h

Dose descriptor:

other: LT50

Effect conc.:

80 ppm

Nominal / measured:

estimated

Conc. based on:

other: Read-across from an analogue

Basis for effect:

mortality

Remarks on result:

other: read-across from an analogue for which LT50 = 12.9 h (95 % CL: 9.9 - 39.4 h)

Details on results:

Based on read across from the analogue camphene, the following results were estimated for the reaction mass:
LT50 (Time to 50% population mortality): 12.9 h at a concentration of 80 ppm.
Mortality of 83 % of beetles at 80 ppm after 24 hours of exposure.

Validity criteria fulfilled:

not applicable

Conclusions:

Based on the read-across approach from the analogue camphene, the lethal time LT50 of reaction mass for Dendroctonus rufipennis (spruce beetle ) was 12.9 hours at 80 ppm, and a mortality of 83 % after 24 hours of exposure was estimated.

Executive summary:

The toxicity of Camphene to Dendroctonus rufipennis (spruce beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus rufipennis (spruce beetle ) was 12.9 hours. Mortality of 83 % of bettles at 80 ppm was observed after 24 hours of exposure. Based on these results, the read-across approach was applied and the lethal time LT50 of reaction mass for Dendroctonus rufipennis (spruce beetle ) was determined to be 12.9 hours at 80 ppm, and a mortality of 83 % after 24 hours of exposure was estimated.

Reference 12

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
The analogue substance alpha pinene which shares the same functional groups with the main components of the multi-constituent substance reaction mass of fenchene and laevo camphene and dextro camphene and laevo alpha pinene also has comparable values for the relevant molecular properties.
See attached the reporting format.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

3.6 h

Dose descriptor:

other: LT50

Effect conc.:

80 ppm

Nominal / measured:

estimated

Conc. based on:

other: Read-across from an analogue

Basis for effect:

mortality

Remarks on result:

other: read-across from an analogue for which LT50= 3.6 h (95 % CL: 2.1 - 6.3 h)

Details on results:

Based on read across from the analogue alpha pinene, the following results were estimated for the reaction mass:
LT50 (Time to 50% population mortality): 3.6 h at a concentration of 80 ppm.
Mortality of 100 % of beetles at 80 ppm after 24 hours of exposure.

Validity criteria fulfilled:

not applicable

Conclusions:

Based on the read-across approach from the analogue alpha pinene, the lethal time LT50 of reaction mass for Dendroctonus simplex (eastern larch beetle ) was 3.6 hours at 80 ppm, and a mortality of 100 % after 24 hours of exposure was estimated.

Executive summary:

The toxicity of alpha pinene to Dendroctonus simplex (eastern larch beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus simplex (eastern larch beetle ) was 3.6 hours. Mortality of 100 % of bettles at 80 ppm was observed after 24 hours of exposure. Based on these results, the read-across approach was applied and the lethal time LT50 of reaction mass for Dendroctonus simplex (eastern larch beetle) was determined to be 3.6 hours at 80 ppm, and a mortality of 100 % after 24 hours of exposure was estimated.

Reference 13

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
The analogue substance camphene which shares the same functional groups with the main components of the multi-constituent substance reaction mass of fenchene and laevo camphene and dextro camphene and laevo alpha pinene also has comparable values for the relevant molecular properties.
See attached the reporting format.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

16.3 h

Dose descriptor:

other: LT50

Effect conc.:

80 ppm

Nominal / measured:

estimated

Conc. based on:

other: Read-across from an analogue

Basis for effect:

mortality

Remarks on result:

other: read-across from an analogue for which LT50 = 16.3 h (95 % CL: 11.2 - 23.2 h)

Details on results:

Based on read across from the analogue camphene, the following results were estimated for the reaction mass:
LT50 (Time to 50% population mortality): 16.3 h at a concentration of 80 ppm.
Mortality of 77 % of beetles at 80 ppm after 24 hours of exposure.

Validity criteria fulfilled:

not applicable

Conclusions:

Based on the read-across approach from the analogue camphene, the lethal time LT50 of reaction mass for Dendroctonus simplex (eastern larch beetle ) was 16.3 hours at 80 ppm, and a mortality of 77 % after 24 hours of exposure was estimated.

Executive summary:

The toxicity of Camphene to Dendroctonus simplex (eastern larch beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus simplex (eastern larch beetle ) was 16.3 hours. Mortality of 77 % of bettles at 80 ppm was observed after 24 hours of exposure. Based on these results, the read-across approach was applied and the lethal time LT50 of reaction mass for Dendroctonus simplex (eastern larch beetle) was determined to be 16.3 hours at 80 ppm, and a mortality of 77 % after 24 hours of exposure was estimated.

Description of key information

Data waiving (exposure considerations): According to the column 2 of REACH Annex IX, the study does not be conducted since direct and indirect exposure of the soil compartment is unlikely.

Supporting study: Read-across approach. camphene: Based on the read-across approach from the analogue camphene, the following results for the reaction mass are estimated: The lethal time (LT50, time to 50% population mortality) for Dendroctonus rufipennis (spruce beetle ) is 12.9 hours. Mortality of 83 % of beetles at 80 ppm is calculated after 24 hours of exposure. The lethal time (LT50, time to 50% population mortality) for Dendroctonus simplex (eastern larch beetle ) is 16.3 hours. Mortality of 77 % of beetles at 80 ppm is calculated after 24 hours of exposure.

Supporting study: Read-across approach. alpha-pinene: Based on the read-across approach from the analogue alpha pinene, the following results for the reaction mass are estimated: The lethal time (LT50, time to 50% population mortality) for Dendroctonus rufipennis (spruce beetle ) is 5.9 hours. Mortality of 100 % of beetles at 80 ppm is calculated after 24 hours of exposure. The lethal time (LT50, time to 50% population mortality) for Dendroctonus simplex (eastern larch beetle ) is 3.6 hours. Mortality of 100 % of beetles at 80 ppm is calculated after 24 hours of exposure.

Supporting study: Read-across approach. L-alpha-pinene (larvicidal assay): Based on the read-across approach from the analogue l-alpha pinene, the following result for the reaction mass is estimated: The 24 h larvicidal LC50 on Aedes albopictus (Asian tiger Mosquito) is estimated to be 72.3 mg/L (95 % CL: 67.79-76.49).

Supporting study: Read-across approach. L-alpha-pinene (repellency test): Based on the read-across approach from the analogue l-alpha pinene, the following result for the reaction mass is estimated: the NOEL for repellent activity against Aedes albopictus (Asian tiger mosquito) is determined to be 0.2 μlcm−2.

Additional information

Data waiving (exposure considerations): According to the column 2 of REACH Annex IX, the study does not be conducted since direct and indirect exposure of the soil compartment is unlikely.

Supporting study: Read-across approach. camphene: The toxicity of Camphene to Dendroctonus rufipennis (spruce beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus rufipennis (spruce beetle ) was 12.9 hours. Mortality of 83 % of bettles at 80 ppm was observed after 24 hours of exposure. Based on these results, the read-across approach was applied and the lethal time LT50 of reaction mass for Dendroctonus rufipennis (spruce beetle ) was determined to be 12.9 hours at 80 ppm, and a mortality of 83 % after 24 hours of exposure was estimated.

Supporting study: Read-across approach. camphene: The toxicity of Camphene to Dendroctonus simplex (eastern larch beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus simplex (eastern larch beetle ) was 16.3 hours. Mortality of 77 % of bettles at 80 ppm was observed after 24 hours of exposure. Based on these results, the read-across approach was applied and the lethal time LT50 of reaction mass for Dendroctonus simplex (eastern larch beetle) was determined to be 16.3 hours at 80 ppm, and a mortality of 77 % after 24 hours of exposure was estimated.

Supporting study: Read-across approach. alpha-pinene: The toxicity of alpha pinene to Dendroctonus rufipennis (spruce beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus rufipennis (spruce beetle ) was 5.9 hours. Mortality of 100 % of bettles at 80 ppm was observed after 24 hours of exposure. Based on these results, the read-across approach was applied and the lethal time LT50 of reaction mass for Dendroctonus rufipennis (spruce beetle ) was determined to be 5.9 hours at 80 ppm, and a mortality of 100 % after 24 hours of exposure was estimated.

Supporting study: Read-across approach. alpha-pinene: The toxicity of alpha pinene to Dendroctonus simplex (eastern larch beetle) was tested by exposing male and female beetles to vapors of test substance. Beetles were exposed to 20, 40, 60 and 80 ppm for 24 hours, using five replicates per dose and a control group of water alone. Mortality was observed and recorded 2, 6, 12, 18 and 24 hours after exposure. Lethal time (LT50, time to 50% population mortality) was determined by probit analysis for each treatment in the beetle assays. The lethal time for Dendroctonus simplex (eastern larch beetle ) was 3.6 hours. Mortality of 100 % of bettles at 80 ppm was observed after 24 hours of exposure. Based on these results, the read-across approach was applied and the lethal time LT50 of reaction mass for Dendroctonus simplex (eastern larch beetle) was determined to be 3.6 hours at 80 ppm, and a mortality of 100 % after 24 hours of exposure was estimated.

Supporting study: L-alpha-pinene (larvicidal assay): A larvicidal bioassay on Aedes albopictus (Asian tiger Mosquito) was conducted for some citrus essential oils and in particular for one of their components L-alpha pinene in accordance to the test method of larval susceptibility as suggested by the World Health Organization (WHO 2005). Twenty late third-to early fourth-instar mosquito larvae were placed in 2 % v/v aqueous solution of DMSO (98 ml of tap water plus 2 ml of DMSO), followed by addition of the tested material solution (10% w/v in DMSO). Four replicates and one negative control (2% v/v in DMSO) were tested. After 24 h exposure the mortality was recorded and the LC50 of the test item, determined by probit analysis, was estimated to be 72.3 mg/L (95 % CL: 67.79-76.49). Based on these results, the read-across approach was applied and the 24 h larvicidal LC50 for the reaction mass was estimated to be 72.3 mg/L (95 % CL: 67.79-76.49).

Supporting study: L-alpha-pinene (repellency test): A repellent activity assay against Aedes albopictus (Asian tiger mosquito) was conducted for some citrus essential oils and in particular for one of their components L-alpha pinene, based on the human landing counts (Coleman et al. 1993; Govere and Durrheim 2006). The test material was applied on filter paper of 24 cm2 total area placed around the opening measuring of 5×5 cm made in a plastic glove, which was inserted for 5 min into the cage containing 100 adult mosquitoes. Two doses of test item (0.2 μlcm−2 and 0.4 μlcm−2), negative control and positive control (N,N-diethyl-3-methylbenzamide (DEET)) were tested and repeated eight times. Based on average number of landings compared with the controls, the NOEL of the test item was determined to be 0.2 μlcm−2. Based on these results, the read-across approach was applied and the NOEL of the reaction mass was determined to be 0.2 μlcm−2.