Registration Dossier

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 November 2014 - 20 November 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted according the OECD 408 guideline and GLP compliance. Fully adequate for assessment.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in Section 13.
Cross-reference
Reason / purpose:
read-across: supporting information
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
13 November 2014 - 20 November 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted according the OECD 408 guideline and GLP compliance. Fully adequate for assessment.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in Section 13.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Qualifier:
according to
Guideline:
other: Commission Regulation (EC) No 440/2008 of 30 May 2008, laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
Identification : Oct-1-ene (#2) CAS 111-66-0
Physical State/Appearance : clear colorless liquid
Purity : 99.9%
Lot Number : 748027
Label : Alpha Olefin C8 (1-Octene) 5L Lot 748027
Date Received : 03 March 2014
Storage Conditions : Room temperature in the dark under nitrogen
Expiry Date : 17 February 2015
Species:
rat
Strain:
other: Wistar Han™:RccHan™:WIST
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories U.K. Ltd., Oxon, UK
- Age at study initiation: six to eight weeks old
- Weight at study initiation: Males: 200 to 242g, Females: 161 to 193g
- Housing: The animals were housed in groups of three or four by sex in solid floor polypropylene cages with stainless steel mesh lids and softwood flake bedding
- Diet (e.g. ad libitum): A pelleted diet (Rodent 2014C Teklad Global Certified Diet, Harlan Laboratories U.K. Ltd., Oxon, UK - free access
- Water (e.g. ad libitum): free access
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 50 ± 20%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hr light/12 hr dark

Experimental Starting Date: 13 November 2014
Experimental Completition Date: 20 November 2015

Justification for specie selection: The rat was selected for this study as it is a readily available rodent species historically used in safety evaluation studies and is acceptable to appropriate regulatory authorities.

Route of administration:
oral: gavage
Vehicle:
other: arachis oil BP
Details on oral exposure:
The test item was administered daily, for ninety consecutive days, by gavage using a stainless steel cannula attached to a disposable plastic syringe. Control animals were treated in an identical manner with 4 mL/kg of Arachis oil BP.

The volume of test and control item administered to each animal was based on the most recent scheduled body weight and was adjusted at weekly intervals.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test item was prepared in Arachis oil BP solution. The stability and homogeneity of the test item formulations were determined by Harlan Laboratories Ltd., Shardlow, UK, Analytical Services. The results of the analytical determination showed that the formulations were stable for at least 20 days. Formulations were prepared and stored at approximately 4 °C in the dark.
The results indicate that the prepared formulations were in the range between 96% and 102% of the nominal concentration confirming the suitability and accuracy of the formulation procedure.
Duration of treatment / exposure:
90 days
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
0 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
100 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
300 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected based on a fourteen day dose range finding study in the rat (Harlan Study Number: 41301411). In this study, a high dosage of 1000 mg/kg bw/day was well tolerated, therefore, the dose 0 (Control), 100, 300 and 1000 mg/kg bw/day have been selected for the OECD 408 study.

Observations and examinations performed and frequency:
Clinical Observations:
- Signs of toxicity
- Ill-health or behavioural change immediately before dosing

Functional Observations:
- Prior to the start of treatment and at weekly intervals thereafter

Behavioural Assessment
- Functional Performance Tests: Motor Activity, Forelimb/Hindlimb Grip Strength
- Sensory Reactivity

Body Weight
- Individual body weights were recorded on Day 1 (prior to dosing), at weekly intervals thereafter and at terminal kill

Food Consumption
- Weekly intervals throughout the study

Water Consumption
- Daily by visual inspection of the water bottles

Ophthalmoscopic Examination
- Pre-treatment and before termination of treatment (during Week 12)

Laboratory Investigations
- End of the study
- Haematology
- Blood Chemistry

Sacrifice and pathology:
Sacrifice: All animals were terminated by intravenous overdose of a suitable barbiturate agent followed by exsanguination.
Pathology
- Necropsy: all animals were subjected to a full external and internal examination.
- Organ Weights (Adrenals, Ovaries, Brain Spleen, Epididymides, Testes, Heart, Thymus, Kidneys, Uterus, Liver)
- Histopathology: Samples of selected tissues were removed from all animals and preserved. All tissues from control and 1000 mg/kg group animals were examined microscopically.
Stomach, lungs and kidney (males only) from 100 and 300 mg/kg groups were examined to clarify potential treatment related findings. In addition the kidneys form male animals were subject to immunohistochemical examination to confirm the presence of alpha-2-microglobulin.
Statistics:
Bartlett’s test
ANOVA
ANCOVA
Williams Test
Student t-test (parametric)
MannWhitney U test
Shirley Test
Dunnett’s
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
not specified
Details on results:
Mortality
- No unscheduled death

Clinical Observations
- 1000 mg/kg bw/day: male and female rats showed increased salivation (males from Day 8 and females from Day 7)
- 1000 mg/kg bw/day: one male and one female show incidences of noisy respiration
- 300 mg/kg bw/day: Slight increased salivation was observed male and females
- 100 mg/kg bw/day: no effects
These findings are considered to be the result of local irritancy of the test item and therefore cannot be considered indicative of true systemic toxicity.

Functional Observations
- Males from all treatment groups and females from 1000 and 300 mg/kg bw group showed a statistically significant reduction in overall activity compared to the control.
- 1000 mg/kg bw/day: males showed a statistically significant reduction in the first fore limb and hind limb grip strength test and a statistically significant increase in the final hind limb grip strength test.
These observation are considered to be not related to the treatment.

- No treatment-related effects were observed in the sensory reactivity parameters.

Body Weight
- No treatment-related effects

Food Consumption
- No treatment-related effects

Water Consumption
- No treatment-related effects

Ophthalmoscopic Examination
- No treatment-related effects

Laboratory Investigations
- No treatment-related effects

Blood Chemistry
- No treatment-related effects

Pathology
Necropsy and Histopathology
A mass surrounding the heart and lungs and a fluid filled vagina was found in one female treated with 1000 mg/kg bw due to an abscess in the heart, granuloma in the lungs and a dilated lumen in the uterus. One control female, one female treated with 100 mg/kg bw/day, two females and one
male treated with 300 mg/kg bw/day and four females treated with 1000 mg/kg bw/day had reddened lungs due to. These findings were considered not related to the treatments.

Organ Weights
- Males (only top dose) and females from all treatment groups showed a statistically significant increase in liver weight both absolute and relative to terminal body weight. No histopathological correlation was observed. Males treated with 1000 mg/kg bw/day showed a statistically significant reduction in epididymides weight both absolute and relative to terminal body weight.
All these finding are not considered related to the treatment.

- Histopathology
Kidney: increased incidence and severity of hyaline droplets and granular casts (consistent with alpha-2u-globulin nephropathy) were evident in males from all treatment groups.
Stomach: Acanthosis, hyperkeratosis and submucosal inflammation was evident in the forestomach of animals of either sex treated with 1000 and 300 mg/kg bw/day.
Lungs: Alveolar macrophages were observed in animals of either sex treated with 1000 mg/kg bw/day and in females treated with 300 mg/kg bw/day. These findings are considered to be a result of local irritation following inhalation of micro-droplets of the test compound (gavage route exposure). All these finding are not considered related to the treatment.



Dose descriptor:
NOEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: Based on treatment related effects in males from all treatment groups and in females treated with 1000 and 300 mg/kg bw/day. The No Observed Effect Level (NOEL) was not established for males.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Based on the fact that the findings in lungs and stomach were not considered to reflect true systemic toxicity. The kidney findings in males are species and sex specific and are not relevant for human health.
Critical effects observed:
not specified
Conclusions:
Based on the results of the study, the No Observed Effect Level (NOEL) was considered to be 100 mg/kg bw/day for females and was not established for males. The No Observed Adverse Effect Level (NOAEL) was established at 1000 mg/kg bw/day for females and males because the findings were either not considered to reflect true systemic toxicity or were not relevant for human health.
Executive summary:

The test material Oct-1-ene CAS 111-66-0 was administrated orally to rats by gavage, at dose levels of 100, 300 and 1000 mg/kg bw/day for 90 consecutive days. The results showed treatment - related effects in males of all dose groups and females treated with 1000 and 300 mg/kg bw.

Pathology examination showed increased incidence and severity of hyaline droplets and granular casts (consistent with alpha-2u-globulin nephropathy) in kidneys of males from all treatment groups. The presence of alpha-2-microglobulin was confirmed histochemically. This finding is species and sex specific and is not considered relevant for human health.

Acanthosis, hyperkeratosis and submucosal inflammation was evident in the forestomach of animals of either sex treated with 1000 and 300 mg/kg bw/day. This finding is a result of local irritation.

Alveolar macrophages were observed in animals of either sex treated with 1000 mg/kg bw/day and in females treated with 300 mg/kg bw/day. This finding is considered to be a result of local irritation following inhalation of micro droplets of the test compound (gavage route exposure) and, therefore, are not considered related to the treatment.

Based on the results of the study, the No Observed Effect Level (NOEL) was considered to be 100 mg/kg bw/day for females and was not established for males. The No Observed Adverse Effect Level (NOAEL) was established at 1000 mg/kg bw/day for females and males because the findings were either not evidence of true systemic toxicity or were not relevant for human health.

Based on the results of this study, the No Observed Effect Level (NOEL) for systemic toxicity was considered to be 1000 mg/kg bw/day.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Qualifier:
according to
Guideline:
other: Commission Regulation (EC) No 440/2008 of 30 May 2008, laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Clear colourless liquid
Details on test material:
- Test Material : Oct-1-ene
- CAS Number: 111-66-0
- Physical State/Appearance : Clear colourless liquid
- Purity : 99.9%
- Lot Number : 748027
- Label: Alpha Olefin C8 (1-Octene) 5L Lot 748027
- Date Received : 03 March 2014
- Storage Conditions : Ambient temperature, in the dark and under nitrogen
- Expiry Date : 17 Feb 2016
Specific details on test material used for the study:
Identification : Oct-1-ene (#2) CAS 111-66-0
Physical State/Appearance : clear colorless liquid
Purity : 99.9%
Lot Number : 748027
Label : Alpha Olefin C8 (1-Octene) 5L Lot 748027
Date Received : 03 March 2014
Storage Conditions : Room temperature in the dark under nitrogen
Expiry Date : 17 February 2015

Test animals

Species:
rat
Strain:
other: Wistar Han™:RccHan™:WIST
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories U.K. Ltd., Oxon, UK
- Age at study initiation: six to eight weeks old
- Weight at study initiation: Males: 200 to 242g, Females: 161 to 193g
- Housing: The animals were housed in groups of three or four by sex in solid floor polypropylene cages with stainless steel mesh lids and softwood flake bedding
- Diet (e.g. ad libitum): A pelleted diet (Rodent 2014C Teklad Global Certified Diet, Harlan Laboratories U.K. Ltd., Oxon, UK - free access
- Water (e.g. ad libitum): free access
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 50 ± 20%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hr light/12 hr dark

Experimental Starting Date: 13 November 2014
Experimental Completition Date: 20 November 2015

Justification for specie selection: The rat was selected for this study as it is a readily available rodent species historically used in safety evaluation studies and is acceptable to appropriate regulatory authorities.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: arachis oil BP
Details on oral exposure:
The test item was administered daily, for ninety consecutive days, by gavage using a stainless steel cannula attached to a disposable plastic syringe. Control animals were treated in an identical manner with 4 mL/kg of Arachis oil BP.

The volume of test and control item administered to each animal was based on the most recent scheduled body weight and was adjusted at weekly intervals.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test item was prepared in Arachis oil BP solution. The stability and homogeneity of the test item formulations were determined by Harlan Laboratories Ltd., Shardlow, UK, Analytical Services. The results of the analytical determination showed that the formulations were stable for at least 20 days. Formulations were prepared and stored at approximately 4 °C in the dark.
The results indicate that the prepared formulations were in the range between 96% and 102% of the nominal concentration confirming the suitability and accuracy of the formulation procedure.
Duration of treatment / exposure:
90 days
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
100 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
300 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected based on a fourteen day dose range finding study in the rat (Harlan Study Number: 41301411). In this study, a high dosage of 1000 mg/kg bw/day was well tolerated, therefore, the dose 0 (Control), 100, 300 and 1000 mg/kg bw/day have been selected for the OECD 408 study.

Examinations

Observations and examinations performed and frequency:
Clinical Observations:
- Signs of toxicity
- Ill-health or behavioural change immediately before dosing

Functional Observations:
- Prior to the start of treatment and at weekly intervals thereafter

Behavioural Assessment
- Functional Performance Tests: Motor Activity, Forelimb/Hindlimb Grip Strength
- Sensory Reactivity

Body Weight
- Individual body weights were recorded on Day 1 (prior to dosing), at weekly intervals thereafter and at terminal kill

Food Consumption
- Weekly intervals throughout the study

Water Consumption
- Daily by visual inspection of the water bottles

Ophthalmoscopic Examination
- Pre-treatment and before termination of treatment (during Week 12)

Laboratory Investigations
- End of the study
- Haematology
- Blood Chemistry

Sacrifice and pathology:
Sacrifice: All animals were terminated by intravenous overdose of a suitable barbiturate agent followed by exsanguination.
Pathology
- Necropsy: all animals were subjected to a full external and internal examination.
- Organ Weights (Adrenals, Ovaries, Brain Spleen, Epididymides, Testes, Heart, Thymus, Kidneys, Uterus, Liver)
- Histopathology: Samples of selected tissues were removed from all animals and preserved. All tissues from control and 1000 mg/kg group animals were examined microscopically.
Stomach, lungs and kidney (males only) from 100 and 300 mg/kg groups were examined to clarify potential treatment related findings. In addition the kidneys form male animals were subject to immunohistochemical examination to confirm the presence of alpha-2-microglobulin.
Statistics:
Bartlett’s test
ANOVA
ANCOVA
Williams Test
Student t-test (parametric)
MannWhitney U test
Shirley Test
Dunnett’s

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
not specified
Details on results:
Mortality
- No unscheduled death

Clinical Observations
- 1000 mg/kg bw/day: male and female rats showed increased salivation (males from Day 8 and females from Day 7)
- 1000 mg/kg bw/day: one male and one female show incidences of noisy respiration
- 300 mg/kg bw/day: Slight increased salivation was observed male and females
- 100 mg/kg bw/day: no effects
These findings are considered to be the result of local irritancy of the test item and therefore cannot be considered indicative of true systemic toxicity.

Functional Observations
- Males from all treatment groups and females from 1000 and 300 mg/kg bw group showed a statistically significant reduction in overall activity compared to the control.
- 1000 mg/kg bw/day: males showed a statistically significant reduction in the first fore limb and hind limb grip strength test and a statistically significant increase in the final hind limb grip strength test.
These observation are considered to be not related to the treatment.

- No treatment-related effects were observed in the sensory reactivity parameters.

Body Weight
- No treatment-related effects

Food Consumption
- No treatment-related effects

Water Consumption
- No treatment-related effects

Ophthalmoscopic Examination
- No treatment-related effects

Laboratory Investigations
- No treatment-related effects

Blood Chemistry
- No treatment-related effects

Pathology
Necropsy and Histopathology
A mass surrounding the heart and lungs and a fluid filled vagina was found in one female treated with 1000 mg/kg bw due to an abscess in the heart, granuloma in the lungs and a dilated lumen in the uterus. One control female, one female treated with 100 mg/kg bw/day, two females and one
male treated with 300 mg/kg bw/day and four females treated with 1000 mg/kg bw/day had reddened lungs due to. These findings were considered not related to the treatments.

Organ Weights
- Males (only top dose) and females from all treatment groups showed a statistically significant increase in liver weight both absolute and relative to terminal body weight. No histopathological correlation was observed. Males treated with 1000 mg/kg bw/day showed a statistically significant reduction in epididymides weight both absolute and relative to terminal body weight.
All these finding are not considered related to the treatment.

- Histopathology
Kidney: increased incidence and severity of hyaline droplets and granular casts (consistent with alpha-2u-globulin nephropathy) were evident in males from all treatment groups.
Stomach: Acanthosis, hyperkeratosis and submucosal inflammation was evident in the forestomach of animals of either sex treated with 1000 and 300 mg/kg bw/day.
Lungs: Alveolar macrophages were observed in animals of either sex treated with 1000 mg/kg bw/day and in females treated with 300 mg/kg bw/day. These findings are considered to be a result of local irritation following inhalation of micro-droplets of the test compound (gavage route exposure). All these finding are not considered related to the treatment.



Effect levels

open allclose all
Dose descriptor:
NOEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: Based on treatment related effects in males from all treatment groups and in females treated with 1000 and 300 mg/kg bw/day. The No Observed Effect Level (NOEL) was not established for males.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Based on the fact that the findings in lungs and stomach were not considered to reflect true systemic toxicity. The kidney findings in males are species and sex specific and are not relevant for human health.

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Based on the results of the study, the No Observed Effect Level (NOEL) was considered to be 100 mg/kg bw/day for females and was not established for males. The No Observed Adverse Effect Level (NOAEL) was established at 1000 mg/kg bw/day for females and males because the findings were either not considered to reflect true systemic toxicity or were not relevant for human health.
Executive summary:

The test material Oct-1-ene CAS 111-66-0 was administrated orally to rats by gavage, at dose levels of 100, 300 and 1000 mg/kg bw/day for 90 consecutive days. The results showed treatment - related effects in males of all dose groups and females treated with 1000 and 300 mg/kg bw.

Pathology examination showed increased incidence and severity of hyaline droplets and granular casts (consistent with alpha-2u-globulin nephropathy) in kidneys of males from all treatment groups. The presence of alpha-2-microglobulin was confirmed histochemically. This finding is species and sex specific and is not considered relevant for human health.

Acanthosis, hyperkeratosis and submucosal inflammation was evident in the forestomach of animals of either sex treated with 1000 and 300 mg/kg bw/day. This finding is a result of local irritation.

Alveolar macrophages were observed in animals of either sex treated with 1000 mg/kg bw/day and in females treated with 300 mg/kg bw/day. This finding is considered to be a result of local irritation following inhalation of micro droplets of the test compound (gavage route exposure) and, therefore, are not considered related to the treatment.

Based on the results of the study, the No Observed Effect Level (NOEL) was considered to be 100 mg/kg bw/day for females and was not established for males. The No Observed Adverse Effect Level (NOAEL) was established at 1000 mg/kg bw/day for females and males because the findings were either not evidence of true systemic toxicity or were not relevant for human health.

Based on the results of this study, the No Observed Effect Level (NOEL) for systemic toxicity was considered to be 1000 mg/kg bw/day.