Oct-2-ene

Endpoint summary

Description of key information

Based on the results of a OECD 408 90-day study on the read-across source oct-1-ene in rats (Envigo 2015), 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.

In an Oral (Gavage) Combined Repeat Dose Toxicity Study with Reproduction/Developmental Toxicity Screening Test in the Rat (OECD 422) using oct-1-ene, the ‘No Observed Effect Level’ (NOEL) for systemic toxicity was considered to be 300 mg/kg bw/day, based on irritation in the forestomach at 1000 mg/kg/day. The NOAEL was however considered to be 1000 mg/kg bw/day because the findings were not evidence of true systemic toxicity.

In a 28-day dermal toxicity study, the read-across source alpha olefin 8 was applied to the shaved skin of six New Zealand white rabbits (3 intact and 3 abraded) at a dose level of 0.2 millilitres for 5 days/week during a 28-day period.

The skin was ranked using a graded system by the study authors ranging from 1 (none) to 6 (severe). The average score for hyperaemia, exfoliation, and scab formation was approximately 2 (considered questionable). There were no other toxicity results reported. Therefore, noLOAEL orNOAEL were identified.

Read-across to Oct-2-ene is claimed as valid basd on the justifications provided for both analogue and category approaches. Therefore the systemic NOAEL for oct-2-ene is considered to be 1000 mg/kg bw/day because the findings of both source studies did not produce evidence of true systemic toxicity.

Guideline repeat dose toxicity studies in rats have been conducted for fourteen members of the higher olefin category, covering C6 to C20-24.The majority of these investigations (27 studies) have used oral (gavage) exposure, with three sub-acute (28-day), nine screening (OECD 421/422), and seven sub-chronic (90-day) studies available for this route. Two sub-acute dermal, two sub-acute inhalation and one sub-chronic inhalation tests, are also available; eight short-term repeat dose range-finding studies are also available. For the oral studies, systemic toxicity findings were typically limited to body weight, liver changes, and effects on clinical chemistry parameters as well as organ weights. Some of the effects observed were adaptive rather than adverse. While most of the studies revealed no systemic toxicity at doses up to 1000 mg/Kg bw/day, a conservative NOAEL for this category was determined to be 100 mg/Kg bw/day, based on minor effects observed with some category members.  

Read-across of these results to Oct-2 -ene is claimed as valid based on the justifications provided for both analogue and category approaches

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

sub-chronic toxicity: oral

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

Read-across between the target substance Oct-2-ene (EC 203-894-2 / CAS 111-67-1) and source substances Octene (EC 246-920-8 / CAS 25377-83-7) and Oct-1-ene (EC 203-893-7 / CAS 111-66-0 / alpha-C8) is based upon the similarity of the chemical structures and their respective physico-chemical properties. The ECHA Read-Across Assessment Framework (RAAF) states that substances with qualitatively similar properties can form the basis of read-across in circumstances where the source and target substances share such similar characteristics.

Octene, Oct-1-ene and Oct-2-ene are linear olefins, each with a carbon chain length of C8. Structurally, the difference between source and target substances is the position of the carbon-carbon double bond. For Oct-1-ene the double bond is at the terminal C1 position (hence, an alpha-olefin), whereas for Oct-2-ene the double bond is at the non-terminal C2 position (hence, an internal olefin). A comparison of the target and source substance properties shows that all substances would be expected to exhibit similar environmental fate, ecotoxicological and mammalian toxicological behaviours. The justification for read-across from source substances Octene and Oct-1-ene to target substance Oct-2-ene is detailed in section 13 (Document name: “Oct-2-ene Read Across Document HOPA”).

Further, Oct-2-ene and Oct-1-ene both fit within the boundaries of the chemical category of higher olefins. Studies conducted by the HOPA consortium on a large range of higher olefin category members (including Oct-1-ene) demonstrated sufficiently similar physico-chemical, environmental fate and toxicological properties to substantiate the basis for read-across. Therefore Oct-2-ene is expected to behave similarly. Justification for inclusion of Oct-2-ene within the boundaries of the higher olefins category, and the relevance of each category member as an analogue substance to Oct-2-ene, is provided in Section 13 (Document name: “HOPA Higher Olefins CJD with Category Matrix Report [rev 1 Sept 2016]”).

Reason / purpose for cross-reference:

read-across: supporting information

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Species:

rat

Route of administration:

oral: gavage

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

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Animals of either sex treated with 1000 and 300 mg/kg bw/day showed episodes of increased salivation during the treatment period. One male treated with 1000 mg/kg bw/day showed incidences of noisy respiration and one female from this treatment group had noisy respiration, decreased respiratory rate and hunched posture on Day 68. No such effects were detected in animals of either sex treated with 100 mg/kg bw/day.

Mortality:

no mortality observed

Description (incidence):

There were no unscheduled deaths on the study.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of treatment on body weight performance for either sex at 100, 300 or 1000 mg/kg bw/day.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no treatment-related effects on food consumption at dosages of 100, 300 or 1000 mg/kg bw/day.

Food efficiency:

no effects observed

Description (incidence and severity):

There were no treatment-related effects on food conversion efficiency at dosages of 100, 300 or 1000 mg/kg bw/day.

Water consumption and compound intake (if drinking water study):

no effects observed

Description (incidence and severity):

There was no treatment-related effect detected on water consumption for either sex at dosages of 100, 300 or 1000 mg/kg bw/day.

Ophthalmological findings:

no effects observed

Description (incidence and severity):

There were no treatment-related ocular effects detected.

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

There were no toxicologically significant effects detected in the hematological parameters examined.

Males from all treatment groups showed a statistically significant increase in mean corpuscular hemoglobin concentration (p<0.05). The majority of individual values were within the normal background range and in the absence of a true dose related response the intergroup differences were considered to be of no toxicological significance.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

There were no toxicologically significant effects detected in the blood chemical parameters examined.

Males treated with 1000 mg/kg bw/day showed a statistically significant increase in alanine aminotransferase and females from this treatment group showed a statistically significant increase in urea (p<0.01). The majority of individual values for these parameters were within the normal background ranges and in the absence of any associated histopathological correlates the intergroup differences were considered to be of no toxicological significance.

Males from all treatment groups showed statistically significant reductions in sodium concentration (p<0.05), potassium concentration (p<0.05) and chloride concentration (p<0.05) and a statistically significant increase in inorganic phosphorus (p<0.05). Males treated with 100 mg/kg bw/day also showed a statistically significant increase in bilirubin (p<0.01). Females treated with 1000 and 300 mg/kg bw/day showed a statistically significant increase in glucose (p<0.05). The majority of individual values for these parameters were within the normal background ranges and in the absence of true dose related responses the intergroup differences were considered to be of no toxicological significance.

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

Behavioral Assessment
There were no treatment-related changes in behavioral parameters measured at dosages of 100, 300 or 1000 mg/kg bw/day.

Functional Performance Tests
There were no toxicologically significant changes in functional performance at dosages of 100, 300 or 1000 mg/kg bw/day.

Males from all treatment groups and females treated with 1000 and 300 mg/kg bw/day showed a statistically significant reduction in overall activity when compared with controls (p<0.05-0.01). Females treated with 1000 and 300 mg/kg bw/day also showed a statistically significant reduction in activity during the final 20% period of observation (p<0.05). In the absence of a true dose related response and/orany supporting clinical observations to suggest an effect of neurotoxicity, the intergroup differences were considered to be of no toxicological significance.

Males treated with 1000 mg/kg bw/day showed a statistically significant reduction (p<0.05) in the first fore limb and hind limb grip strength test and a statistically significant increase in the final hind limb grip strength test. In the absence of a consistent effect or any supporting clinical observations to suggest an effect of neurotoxicity, the intergroup differences were considered to be of no toxicological significance.

Sensory Reactivity Assessments
There were no treatment-related changes in sensory reactivity at dosages of 100, 300 or 1000 mg/kg bw/day.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

There were no toxicologically significant effects detected in the organ weights measured.

Males treated 1000 mg/kg bw/day and females from all treatment groups showed a statistically significant increase in liver weight both absolute and relative to terminal body weight (p<0.01; males, p<0.05; females). Although a number of the individual values were outside of the normal background range, no true dose related response for females was evident. In the absence of any associated histopathological correlates, the intergroup differences were therefore considered of no toxicological importance.

Males treated with 1000 mg/kg bw/day showed a statistically significant reduction in epididymides weight both absolute and relative to terminal body weight (p<0.05). All of the individual values were within the normal background range and in the absence of any associated histopathological correlates the intergroup difference was considered to be of no toxicological significance.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

There were no toxicologically significant macroscopic abnormalities detected.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

not specified

Key result

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.

Key result

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.

Key result

Critical effects observed:

no

Conclusions:

Based on the results of a OECD 408 90-day study on oct-1-ene in rats (Envigo 2015), 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.

In an Oral (Gavage) Combined Repeat Dose Toxicity Study with Reproduction/Developmental Toxicity Screening Test in the Rat (OECD 422) using oct-1-ene, the ‘No Observed Effect Level’ (NOEL) for systemic toxicity was considered to be 300 mg/kg bw/day, based on irritation in the forestomach at 1000 mg/kg/day. The NOAEL was however considered to be 1000 mg/kg bw/day because the findings were not evidence of true systemic toxicity.

Read-across to Oct-2-ene is claimed as valid based on the justifications provided for both analogue and category approaches. Therefore the systemic NOAEL for oct-2-ene is considered to be 1000 mg/kg bw/day because the findings of both source studies did not produce evidence of true systemic toxicity.

Reference 2

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

04 July 2013 - 06 March 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study follows the OECD 422 Guidelines and it is GLP compliant. It is classified as reliable without restriction.

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in Section 13.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

yes

Remarks:

The deviations have no adverse impact on the study

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification : Oct-1-ene CAS 111-66-0
Description : clear colorless liquid
Purity : >98%
Batch Number : 719238
Label : Alpha Olefin C8 Lot 719238
Date Received : 29 April 2013
Storage Conditions : room temperature in the dark under nitrogen
Expiry Date : 29 April 2014

No correction for purity was made.

Oct-1-ene, CAS# 111-66-0 used in this study was a typical production sample, according to the details included in the boundary composition in IUCLID section 1.2. Oct-1-ene was chosen in the Higher Olefins category testing strategy because it represents a substance with high alpha olefin content (category range 0 - 98%). Please see the testing strategy attached in section 13 for further details.

Species:

rat

Strain:

other: Wistar Han™:RccHan™:WIST

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories U.K. Ltd., Blackthorn, Bicester, Oxon, UK.
- Age at study initiation: 12 weeks old
- Weight at study initiation: males: 312 to 357g; females: 194 to 234g
- Fasting period before study:
- Housing:
Beginning: all animals were housed in groups of four in solid floor polypropylene cages with stainless steel mesh lids and softwood flake bedding (Datesand Ltd., Cheshire, UK).
Pairing phase: animals were transferred to polypropylene grid floor cages suspended over trays lined with absorbent paper on a one male: one female basis within each dose group.
Evidence of successful mating: the males were returned to their original cages and mated females were housed individually during gestation and lactation in solid floor polypropylene cages with stainless steel mesh lids and softwood flakes.
- Diet (e.g. ad libitum): Pelleted diet (Rodent 2018C Teklad Global Certified Diet, Harlan Laboratories U.K. Ltd., Oxon, UK.)
- Water (e.g. ad libitum): free access
- Acclimation period: eight days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2 °C
- Humidity (%): 55 ± 15% respectively
- Air changes (per hr): at least fifteen air changes per hour
The in-life phase of the study was conducted between 04 July 2013 (first day of treatment) and 21 August 2013 (final day of necropsy).
- Photoperiod (hrs dark / hrs light): 12 hr dark, 12 hr light

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Mixing appropriate amounts with (Type of food): the test item was prepared at the appropriate concentrations as a solution in Arachis oil BP.

VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 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.
Storage vehicle: approximately 4 °C

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Formulations were therefore prepared weekly and stored at approximately 4 °C in the dark under nitrogen. The analytical results indicate that the prepared formulations were within ± 4% of the nominal concentration.

Duration of treatment / exposure:

The test item was administered by gavage to three groups (12/sex) for up to eight weeks (including a two-week pre-pairing phase, pairing, gestation and early lactation for females).

Frequency of treatment:

Daily

Dose / conc.:

0 mg/kg bw/day (actual dose received)

Remarks:

Control

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

twelve male and twelve female Wistar Han™:RccHan™:WIST strain rats

Control animals:

yes, concurrent vehicle

Details on study design:

Twelve male and twelve female animals per dose group were treated daily (except for females during parturition where applicable).

The first day of dosing was designated as Day 1 of the study.

Prior to the start of treatment and once weekly thereafter, all animals were observed for signs of functional/behavioural toxicity.

On Day 15, animals were paired on a 1 male: 1 female basis within each dose group for a maximum of fourteen days.

Following evidence of mating (designated as Day 0 post coitum) the males were returned to their original cages and females were transferred to individual cages.

On completion of the pairing phase (during Week 6), five selected males per dose group were evaluated for functional/sensory responses to various stimuli.

Pregnant females were allowed to give birth and maintain their offspring until Day 5 post partum. Litter size, offspring weight and sex, surface righting and clinical signs were also recorded during this period.

At Day 4 post partum, five selected females per dose group were evaluated for functional/sensory responses to various stimuli.

Blood samples were taken from five males from each dose group for hematological and blood chemical assessments on Day 42. The male dose groups were killed and examined macroscopically on Day 43.

Blood samples were taken from five randomly selected females from each dose group for hematological and blood chemical assessment on Day 4 post partum. At Day 5 post partum, all females and surviving offspring were killed and examined macroscopically. Any female which did not produce a pregnancy was also killed and examined macroscopically.

Observations and examinations performed and frequency:

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: All animals were examined for overt signs of toxicity, ill-health and behavioural change
immediately before dosing, soon after dosing, and one hour after throughout the treatment period (except for females during parturition where applicable).

BODY WEIGHT: Yes
- Individual body weights were recorded on Day 1 (prior to dosing) and then weekly for males until termination and weekly for females until mating was evident. Body weights were then recorded for females on Days 0, 7, 14 and 20 post coitum, and on Days 1 and 4 post partum. Body weights were also recorded at terminal kill.

FOOD CONSUMPTION:
- Pre-pairing period, weekly food consumption was recorded for each cage of adults and males after the mating phase.
- Females showing evidence of mating: food consumption was recorded for the periods covering post coitum Days 0-7, 7-14 and 14-20.
- Females with live litters: food consumption was recorded on Days 1 and 4 post partum.
- Food efficiency (the ratio of body weight change/dietary intake) was calculated retrospectively for males throughout the study period (with the exception of the mating phase) and for females during the pre-pairing phase. Due to offspring growth and milk production, food efficiency
could not be accurately calculated for females during gestation and lactation.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Water intake was measured daily during the pre-pairing phase of the study

BEHAVIORAL, FUNCIONAL OBSERVATIONS and SENSORY REACTIVITIES were observed and recorded.

Sacrifice and pathology:

Pathology Females: Uterus (for signs of implantation and the number of uterine implantations in each horn), the corpora lutea.All adult animals and offspring: full external and internal examination. All macroscopic abnormalities were recorded. - Organ Weights- Histopathology: Tissues from 5 selected male and female animals/group were preserved. Histopathological examination was undertaken on tissues from control and high dose animals. Since there were indications of possible treatment-related changes in the forestomach, examination was subsequently extended to include similarly prepared sections of the stomach from animals in the low and intermediate groups.

Statistics:

Data were analysed using the decision tree from the Provantis Tables and Statistics Module. The homogeneity of variance from mean values was analysed using Bartlett’s test. Intergroup variance were assessed using suitable ANOVA, or if required, ANCOVA with appropriate covarities. Any transformed data were analysed to find the lowest treatment level that showed a significant effect using the Williams Test for parametric data or the Shirley Test for nonparametric data. If no dose response was found but the data shows non-homogeneity of means, the data were analysed by a stepwise Dunnett’s (parametric) or Steel (non-parametric) test to determine significant difference from the control group. Where the data were unsuitable for these analyses, pair-wise tests was performed using the Student t-test (parametric) or the MannWhitney U test (non-parametric).

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Animals of either sex treated with 1000 mg/kg bw/day showed increased salivation from Day 1 (males) and Day 3 (females) onwards. Males treated with 300 mg/kg bw/day also showed episodes of increased salivation between days 16 and 29. No such effects were detected infemales treated with 300 mg/kg bw/day or animals of either sex treated with 100 mg/kg bw/day. One male treated with 1000 mg/kg bw/day, one female treated with 1000 mg/kg bw/day and three control females had generalised fur loss during the treatment period. Observations of this nature are commonly observed in group housed animals and is considered not to be of toxicological significance.

Mortality:

no mortality observed

Description (incidence):

There were no unscheduled deaths.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no treatment related effects detected in body weight development. Statistical analysis of the data did not reveal any significant intergroup differences.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No adverse effects on dietary intake were noted for males during the study or for females during the pre-pairing, gestation or lactation phases of the study. Statistical analysis of female data during gestation and lactation did not reveal any significant intergroup differences.

Food efficiency:

no effects observed

Water consumption and compound intake (if drinking water study):

effects observed, treatment-related

Description (incidence and severity):

Animals of either sex treated with 1000 mg/kg bw/day showed an increase in overall water consumed when compared to the control group.

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No toxicologically significant effects were detected in the haematological parameters examined.

The following intergroup differences were detected however they were considered not to be toxicologically significant. Females treated with 1000 mg/kg bw/day showed an increase in hemoglobin levels whilst males from this treatment group showed a reduction in activated partial thromboplastin time. The majority of individual values were within the normal background range for these parameters. Females from all treatment groups showed an increase in erythrocyte count however a true dose
related response was not evident and all of the individual values were within the normal background range for this parameter.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

No toxicologically significant effects were detected in the blood chemical parameters examined.

The following intergroup differences were detected however they were considered not to be toxicologically significant. Males treated with 1000 and 300 mg/kg bw/day showed a reduction in alkaline phosphatase. Females treated with 1000 mg/kg bw/day showed an increase in albumin. All of the individual values were within the normal background range for these parameters. Males from all treatment groups showed an increase in creatinine however a true dose related response was not evident and the majority of individual values were within the normal background range for this parameter.

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

Behavioral Assessments
Weekly open field arena observations did not reveal any treatment-related effects for treated animals when compared to controls. All inter and intra group differences in behavioural scores were considered to be a result of normal variation for rats of the strain and age used, and the differences were of no toxicological importance.

Functional Performance Tests
There were no toxicologically significant changes in functional performance. Males from all treatment groups showed a statistically significant increase (p<0.05) in mean forelimb grip strength whilst females from all treatment groups showed a statistically significant (p<0.05) reduction in overall activity. The intergroup differences for males were confined to one out of the three tests and in the absence of a true dose related response in either sex was considered not to be of toxicological importance. Males treated with 1000 mg/kg bw/day showed a statistically significant reduction (p<0.05) in overall activity. In the absence of any associated clinical signs to suggest a neurotoxic effectthe intergroup difference was considered not to be of toxicological importance.

Sensory Reactivity Assessments
There were no treatment-related changes in sensory reactivity.

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

No toxicologically significant effects were detected in the organ weights measured.

Males treated with 1000 mg/kg bw/day showed an increase in liver and kidney weight both absolute and relative to terminal body weight. Males from all treatment groups also showed an increase in absolute and relative adrenal weight. Females treated with 1000 mg/kg bw/day showed an increase in absolute and relative thyroid weight. The majorityof individual values for adrenal, kidney and thyroid weights were within the normal background ranges. Although the majority of liver weights were outside the background range, in the absence of any associated histology correlates all the intergroup differenceswere considered not to be of toxicological significance.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

One female treated with 1000 mg/kg bw/day had a thickened stomach at necropsy. This female showed epithelial hyperplasia in the stomach at microscopic examination. No such effects were detected in males treated with 1000 mg/kg bw/day or animals of either sex treated with 300 or 100 mg/kg bw/day. One male treated with 1000 mg/kg bw/day had enlarged and mottled kidneys. One female treated with 300 mg/kg bw/day had reddened lungs at necropsy. In the absence of any histology correlates, the
intergroup differences were considered not to be of toxicological importance.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Stomach
Epithelial hyperplasia in the forestomach was evident in four out of the five males examined and in three out of the five females examined at 1000 mg/kg bw/day. Of the affected animals, two males and two females had minimal severity, two males had moderate severity and one female had slight severity. These findings detected in the forestomach were consistent with local irritation. Humans do not have a rat forestomach counterpart therefore the macroscopic stomach changes detected have limited relevance to human toxicity. No such effects were detected in animals of either sex treated with 300 or 100 mg/kg bw/day.

Other effects:

no effects observed

Details on results:

Clinical Signs
Animals of either sex treated with 1000 mg/kg bw/day showed increased salivation from Day 1 (males) and Day 3 (females) onwards. Males treated with 300 mg/kg bw/day also showed episodes of increased salivation between days 16 and 29. No such effects were detected infemales treated with 300 mg/kg bw/day or animals of either sex treated with 100 mg/kg bw/day. One male treated with 1000 mg/kg bw/day, one female treated with 1000 mg/kg bw/day and three control females had generalised fur loss during the treatment period. Observations of this nature are commonly observed in group housed animals and is considered not to be of toxicological significance.

Water Consumption
Animals of either sex treated with 1000 mg/kg bw/day showed an increase in overall water consumed when compared to the control group.

Hematology
No toxicologically significant effects were detected in the haematological parameters examined. The following intergroup differences were detected however they were considered not to be toxicologically significant. Females treated with 1000 mg/kg bw/day showed an increase in hemoglobin levels whilst males from this treatment group showed a reduction in activated partial thromboplastin time. The majority of individual values were within the normal background range for these parameters. Females from all treatment groups showed an increase in erythrocyte count however a true dose related response was not evident and all of the individual values were within the normal background range for this parameter.

Clinical Biochemistry
No toxicologically significant effects were detected in the blood chemical parameters examined.

The following intergroup differences were detected however they were considered not to be toxicologically significant. Males treated with 1000 and 300 mg/kg bw/day showed a reduction in alkaline phosphatase. Females treated with 1000 mg/kg bw/day showed an increase in albumin. All of the individual values werewithin the normal background range for these parameters. Males from all treatment groups showed an increase in creatinine however a true dose related response was not evident
and the majority of individual values were within the normal background range for this parameter.

Behavior: Functional Performance Tests
There were no toxicologically significant changes in functional performance. Males from all treatment groups showed a statistically significant increase (p<0.05) in mean forelimb grip strength whilst females from all treatment groups showed a statistically significant (p<0.05) reduction in overall activity. The intergroup differences for males were confined to one out of the three tests and in the absence of a true dose related response in either sex was considered not to be of toxicological importance. Males treated with 1000 mg/kg bw/day showed a statistically significant reduction (p<0.05) in overall activity. In the absence of any associated clinical signs to suggest a neurotoxic effectthe intergroup difference was considered not to be of toxicological importance.

Organ Weights
No toxicologically significant effects weredetected in the organ weights measured.

Males treated with 1000 mg/kg bw/day showed an increase in liver and kidney weight both absolute and relative to terminal body weight. Males from all treatment groups also showed an increase in absolute and relative adrenal weight. Females treated with 1000 mg/kg bw/day showed an increase in absolute and relative thyroid weight. The majorityof individual values for adrenal, kidney and thyroid weights were within the normal background ranges. Although the majority of liver weights were outside the background range, in the absence of any associated histology correlates all the intergroup differenceswere considered not to be of toxicological significance.

Necropsy
One female treated with 1000 mg/kg bw/day had a thickened stomach at necropsy. This female showed epithelial hyperplasia in the stomach at microscopic examination. No such effects were detected in males treated with 1000 mg/kg bw/day or animals of either sex treated with 300 or 100 mg/kg bw/day. One male treated with 1000 mg/kg bw/day had enlarged and mottled kidneys. One female treated with 300 mg/kg bw/day had reddened lungs atnecropsy. In the absence of any histology correlates,
the intergroup differences were considered not to beof toxicological importance.

Histopathology
Stomach - Epithelial hyperplasia in the forestomach was evident in four out of the five males examined and in three out of the five females examined at 1000 mg/kg bw/day. Of the affected animals, two males and two females had minimal severity, two males had moderate severity and one female had slight severity. These findings detected in the forestomach were consistent with local irritation. No such effects were detected in animals of either sex treated with 300 or 100 mg/kg bw/day.

Key result

Dose descriptor:

NOEL

Effect level:

300 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Based on irritant effects in the fore stomach

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: The findings were not considered to reflect true systemic toxicity

Key result

Critical effects observed:

no

Table 1. Group Mean Functional Test Values and Standard Deviations - Males
Group (sex)		Test 1 Forelimb (g)	Test 1 Hindlimb (g)	Test 2 Forelimb (g)	Test 2 Hindlimb (g)	Test 3 Forelimb (g)	Test 3 Hindlimb (g)	Overall Activity	Overall Mobile	Last 20% Activity	Last 20% Mobile
1 (M)	Mean	853.0	408.6	536.8	281.6	783.2	377.6	383.2	0.4	19.4	0.2n
	S.D.	168.8	91.6	191.1	86.5	264.2	116.1	157.6	0.5	24.5	0.4
	N	5	5	5	5	5	5	5	5	5	5
2 (M)	Mean	909.8	436.0	934.8*	505.2	925.8	515.8	367.2	0.8	2.4	0.0n
	S.D.	149.8	109.3	251.6	132.7	223.0	218.8	85.5	1.3	2.9	0.0
	N	5	5	5	5	5	5	5	5	5	5
3 (M)	Mean	964.0	349.8	824.6*	482.6	950.6	560.0	285.0	1.0	11.6	0.0n
	S.D.	227.4	102.7	198.9	223.2	154.9	145.4	94.1	1.4	24.8	0.0
	N	5	5	5	5	5	5	5	5	5	5
4 (M)	Mean	887.4	375.4	795.0*	425.4	815.4	442.0	220.6*	0.2	31.2	0.0n
	S.D.	176.1	101.9	162.0	137.3	143.9	54.6	106.1	0.4	57.7	0.0
	N	5	5	5	5	5	5	5	5	5	5

General Footnote: Unit = Time (seconds) for Motor Activity Assessments

Dose Levels: Group 1 - 0(Control); Group 2 - 100 mg/kg bw/day; Group 3 - 300 mg/kg bw/day; Group 4 - 1000 mg/kg bw/day

* Significantly different from control group p<0.05

n Data not appropriate for statistical analysis

 

Table 2. Group Mean Functional Test Values and Standard Deviations - Females
Group (sex)		Test 1 Forelimb (g)	Test 1 Hindlimb (g)	Test 2 Forelimb (g)	Test 2 Hindlimb (g)	Test 3 Forelimb (g)	Test 3 Hindlimb (g)	Overall Activity	Overall Mobile	Last 20% Activity	Last 20% Mobile
1 (F)	Mean	847.8	478.0	960.0	502.6	877.4	450.8	693.2	5.2	77.4	1.2n
	S.D.	163.5	131.8	87.0	139.4	132.6	87.1	149.6	4.3	49.3	1.8
	N	5	5	5	5	5	5	5	5	5	5
2 (F)	Mean	859.8	468.6	745.0	422.4	748.0	476.6	513.2*	5.2	87.2	1.8n
	S.D.	148.0	153.0	220.6	154.4	201.0	98.7	45.0	3.0	73.5	2.2
	N	5	5	5	5	5	5	5	5	5	5
3 (F)	Mean	803.4	383.4	679.2	309.2	827.8	369.0	586.2*	4.2	67.6	0.4n
	S.D.	220.7	126.6	99.9	73.8	262.4	99.3	77.7	3.8	62.1	0.9
	N	5	5	5	5	5	5	5	5	5	5
4 (F)	Mean	809.8	463.2	763.8	405.2	718.8	377.2	519.8*	3.4	69.0	0.0n
	S.D.	184.3	96.5	252.6	118.8	191.1	120.5	115.5	1.5	74.1	0.0
	N	5	5	5	5	5	5	5	5	5	5

General Footnote: Unit = Time (seconds) for Motor Activity Assessments

Dose Levels: Group 1 - 0(Control); Group 2 - 100 mg/kg bw/day; Group 3 - 300 mg/kg bw/day; Group 4 - 1000 mg/kg bw/day

* Significantly different from control group p<0.05

n Data not appropriate for statistical analysis

Table 3. Group Mean Hematological Values - Males
Group		APTT (Seconds)
Group 1 (0 – Control)	Mean	16.14
	S.D.	0.79
	N	10
Group 2 (100 mg/Kg bw/day)	Mean	15.50
	S.D.	2.21
	N	10
Group 3 (300 mg/Kg bw/day)	Mean	14.26
	S.D.	2.71
	N	10
Group 4 (1000 mg/Kg bw/day)	Mean	13.04*
	S.D.	1.38
	N	10

* Significantly different from control group p<0.05

 

Table 4. Group Mean Hematological Values - Females
Group		Hb (g/dL)	RBC (10^12/L)
Group 1 (0 – Control)	Mean	12.76	6.778
	S.D.	0.38	0.318
	N	5	5
Group 2 (100 mg/Kg bw/day)	Mean	13.26	7.344*
	S.D.	0.53	0.383
	N	5	5
Group 3 (300 mg/Kg bw/day)	Mean	13.28	7.212*
	S.D.	0.96	0.426
	N	5	5
Group 4 (1000 mg/Kg bw/day)	Mean	13.66*	7.278*
	S.D.	0.48	0.272
	N	5	5

* Significantly different from control group p<0.05

Table 5. Group Mean Blood Chemical Values - Males
Group (sex)		AP (IU/L)	Creat (mg/dL)
Group 1 (0 – Control)	Mean	244.2	0.660
	S.D.	77.3	0.046
	N	5	5
Group 2 (100 mg/Kg bw/day)	Mean	212.6	0.756*
	S.D.	48.5	0.071
	N	5	5
Group 3 (300 mg/Kg bw/day)	Mean	166.4*	0.888*
	S.D.	44.4	0.288
	N	5	5
Group 4 (1000 mg/Kg bw/day)	Mean	131.0**	0.714*
	S.D.	42.3	0.043
	N	5	5

* Significantly different from control group p<0.05

** Significantly different from control group p<0.01

 

Table 6. Group Mean Blood Chemical Values - Females
Group		Albumin (g/dL)
Group 1 (0 – Control)	Mean	3.38
	S.D.	0.19
	N	5
Group 2 (100 mg/Kg bw/day)	Mean	3.06
	S.D.	0.96
	N	5
Group 3 (300 mg/Kg bw/day)	Mean	3.66
	S.D.	0.15
	N	5
Group 4 (1000 mg/Kg bw/day)	Mean	3.76*
	S.D.	0.23
	N	5

* Significantly different from control group p<0.05

Table 7. Summary Incidence of Necropsy Findings - Males
	Males
	0 Control	100 mg/Kg bw/day	300 mg/Kg bw/day	1000 mg/Kg bw/day
Number of animals examined	12	12	12	12
Kidneys
Submitted	(12)	(12)	(12)	(12)
No Visible Lesions	12	12	12	11
Enlarged	0	0	0	1
Mottled Appearance	0	0	0	1

 

Table 8. Summary Incidence of Necropsy Findings - Females
	Females
	0 Control	100 mg/Kg bw/day	300 mg/Kg bw/day	1000 mg/Kg bw/day
Number of animals examined	12	12	12	12
Lungs (With Bronchi)
Submitted	(12)	(12)	(12)	(12)
No Visible Lesions	12	10	11	12
Reddened	0	2	1	0
Stomach
Submitted	(12)	(12)	(12)	(12)
No Visible Lesions	12	12	12	11
Thickened	0	0	0	1

Table 9. Group Mean Organ Weights with Corresponding Relative (% of Body Weight) Organ Weights
		Males				Females
		0 Control	100 mg/Kg bw/day	300 mg/Kg bw/day	1000 mg/Kg bw/day	0 Control	100 mg/Kg bw/day	300 mg/Kg bw/day	1000 mg/Kg bw/day
Adrenals	Mean (g)	0.05670	0.07550*	0.07714*	0.07736*
	S.D.	0.00630	0.01491	0.01091	0.01645
	N	5	5	5	5
	Mean (%)	0.014	0.018*	0.019*	0.019*
	S.D.	0.002	0.003	0.003	0.003
	N	5	5	5	5
Kidneys	Mean (g)	2.31488	2.37080	2.51686	2.69416*
	S.D.	0.26047	0.24924	0.17059	0.26830
	N	5	5	5	5
	Mean (%)	0.569	0.583	0.602	0.656*
	S.D.	0.031	0.088	0.041	0.074
	N	5	5	5	5
Liver	Mean (g)	13.0518	12.8470	13.4033	15.5767**
	S.D.	1.62501	0.47524	1.39447	0.31896
	N	5	5	5	5
	Mean (%)	3.205	3.151	3.196	3.792**
	S.D.	0.266	0.215	0.211	0.209
	N	5	5	5	5
Thyroid/Parathyroid	Mean (g)					0.01646	0.01892	0.01566	0.02124**
	S.D.					0.00178	0.00255	0.00217	0.00313
	N					5	5	5	5
	Mean (%)					0.006	0.007	0.006	0.008**
	S.D.					0.001	0.001	0.001	0.002
	N					5	5	5	5

* Significantly different from control group p<0.05

** Significantly different from control group p<0.01

Conclusions:

The ‘No Observed Effect Level’ (NOEL) for systemic toxicity was considered to be 300 mg/kg bw/day, based on irritation in the forestomach at 1000 mg/kg/day. The NOAEL was however considered to be 1000 mg/kg bw/day because the findings were not evidence of true systemic toxicity.

Executive summary:

An Oral (Gavage) Combined Repeat Dose Toxicity Study with Reproduction/Developmental Toxicity Screening Test in the Rat (OECD 422) was performed on the test material Oct-1 -ene CAS 111 -66 -0.

After eight weeks treatment clinical signs were detected in animals of either sex treated with 1000 mg/kg bw/day and in males treated with 300 mg/kg bw/day. Increased salivation was evident throughout the treatment period, but no alteration of the physical condition was observed. No difference between animals treated and controls were detected in body weight development and food consumption. In contrast, the water consumption was increased in animals of either sex treated with 1000 mg/kg bw/day. Increased salivation and increased water consumption might be related to unpalatability problems and irritancy of the stomach. Microscopic investigations of the stomachs showed epithelial hyperplasia in animals of either sex treated with 1000 mg/kg bw/day and thickening of the stomach in one female treated with 1000 mg/kg bw/day at necropsy. The findings are considered a result of local irritation rather than any adverse systemic toxicity of the test item. All parameters related to blood and chemistry examinations and reproductive system showed no treatment-related effects.

In view of these results, the NOAEL for systemic toxicity was considered to be 1000 mg/kg bw/day.

Reference 3

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.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Qualifier:

according to guideline

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. QA statement)

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

Oct-1-ene, CAS# 111-66-0 used in this study was a typical production sample, according to the details included in the boundary composition in IUCLID section 1.2. Oct-1-ene was chosen in the Higher Olefins category testing strategy because it represents a substance with high alpha olefin content (category range 0 - 98%). Please see the testing strategy attached in section 13 for further details.

Species:

rat

Strain:

other: Wistar Han™:RccHan™:WIST

Sex:

male/female

Details on test animals or test system 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

Description (incidence and severity):

Animals of either sex treated with 1000 and 300 mg/kg bw/day showed episodes of increased salivation during the treatment period. One male treated with 1000 mg/kg bw/day showed incidences of noisy respiration and one female from this treatment group had noisy respiration, decreased respiratory rate and hunched posture on Day 68. No such effects were detected in animals of either sex treated with 100 mg/kg bw/day.

Mortality:

no mortality observed

Description (incidence):

There were no unscheduled deaths on the study.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There was no effect of treatment on body weight performance for either sex at 100, 300 or 1000 mg/kg bw/day.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no treatment-related effects on food consumption at dosages of 100, 300 or 1000 mg/kg bw/day.

Food efficiency:

no effects observed

Description (incidence and severity):

There were no treatment-related effects on food conversion efficiency at dosages of 100, 300 or 1000 mg/kg bw/day.

Water consumption and compound intake (if drinking water study):

no effects observed

Description (incidence and severity):

There was no treatment-related effect detected on water consumption for either sex at dosages of 100, 300 or 1000 mg/kg bw/day.

Ophthalmological findings:

no effects observed

Description (incidence and severity):

There were no treatment-related ocular effects detected.

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

There were no toxicologically significant effects detected in the hematological parameters examined.

Males from all treatment groups showed a statistically significant increase in mean corpuscular hemoglobin concentration (p<0.05). The majority of individual values were within the normal background range and in the absence of a true dose related response the intergroup differences were considered to be of no toxicological significance.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

There were no toxicologically significant effects detected in the blood chemical parameters examined.

Males treated with 1000 mg/kg bw/day showed a statistically significant increase in alanine aminotransferase and females from this treatment group showed a statistically significant increase in urea (p<0.01). The majority of individual values for these parameters were within the normal background ranges and in the absence of any associated histopathological correlates the intergroup differences were considered to be of no toxicological significance.

Males from all treatment groups showed statistically significant reductions in sodium concentration (p<0.05), potassium concentration (p<0.05) and chloride concentration (p<0.05) and a statistically significant increase in inorganic phosphorus (p<0.05). Males treated with 100 mg/kg bw/day also showed a statistically significant increase in bilirubin (p<0.01). Females treated with 1000 and 300 mg/kg bw/day showed a statistically significant increase in glucose (p<0.05). The majority of individual values for these parameters were within the normal background ranges and in the absence of true dose related responses the intergroup differences were considered to be of no toxicological significance.

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

Behavioral Assessment
There were no treatment-related changes in behavioral parameters measured at dosages of 100, 300 or 1000 mg/kg bw/day.

Functional Performance Tests
There were no toxicologically significant changes in functional performance at dosages of 100, 300 or 1000 mg/kg bw/day.

Males from all treatment groups and females treated with 1000 and 300 mg/kg bw/day showed a statistically significant reduction in overall activity when compared with controls (p<0.05-0.01). Females treated with 1000 and 300 mg/kg bw/day also showed a statistically significant reduction in activity during the final 20% period of observation (p<0.05). In the absence of a true dose related response and/orany supporting clinical observations to suggest an effect of neurotoxicity, the intergroup differences were considered to be of no toxicological significance.

Males treated with 1000 mg/kg bw/day showed a statistically significant reduction (p<0.05) in the first fore limb and hind limb grip strength test and a statistically significant increase in the final hind limb grip strength test. In the absence of a consistent effect or any supporting clinical observations to suggest an effect of neurotoxicity, the intergroup differences were considered to be of no toxicological significance.

Sensory Reactivity Assessments
There were no treatment-related changes in sensory reactivity at dosages of 100, 300 or 1000 mg/kg bw/day.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

There were no toxicologically significant effects detected in the organ weights measured.

Males treated 1000 mg/kg bw/day and females from all treatment groups showed a statistically significant increase in liver weight both absolute and relative to terminal body weight (p<0.01; males, p<0.05; females). Although a number of the individual values were outside of the normal background range, no true dose related response for females was evident. In the absence of any associated histopathological correlates, the intergroup differences were therefore considered of no toxicological importance.

Males treated with 1000 mg/kg bw/day showed a statistically significant reduction in epididymides weight both absolute and relative to terminal body weight (p<0.05). All of the individual values were within the normal background range and in the absence of any associated histopathological correlates the intergroup difference was considered to be of no toxicological significance.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

There were no toxicologically significant macroscopic abnormalities detected.

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 at necropsy. At microscopic examination, the treated animals were shown to have congestion in the lungs. Although there was an increased incidence/severity of congestion in the lungs the intergroup differences were considered to be within the normal range of background alterations that are seen in untreated animals of this age and strain and therefore was considered an incidental finding, not related to the treatments.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

The following treatment related microscopic abnormalities were detected:

Kidneys:increased incidence and severity of hyaline droplets and granular casts (consistent with alpha-2u-globulin nephropathy) were evident in males from all treatment groups. Although the findings of granular casts in the male kidney may be considered to represent an adverse effect of the test item, immunohistochemical staining demonstrated that these changes were correlated to accumulation of alpha 2u-globulin and formation of hyaline droplets, which is unique to the male rat. This finding is commonly observed in male rats following treatment with some hydrocarbons and is not predictive of any adverse effects in humans.

Granular casts may be considered to represent an adverse effect of the test item, however immunohistochemical staining demonstrated that these changes within the kidneys was correlated to the same condition as hyaline droplets therefore the kidney changes were considered to be of limited toxicological significance.

Stomach: Acanthosis, hyperkeratosis, ulceration and submucosal inflammation was evident in the forestomach of animals of either sex treated with 1000 and 300 mg/kg bw/day. The stomach changes identified in 1000 and 300 mg/kg bw/day animals were considered to be a result of local irritation of the test item rather than a true effect of systemic toxicity.

Lungs:increased incidence and severity of alveolar macrophages were evident in animals of either sex treated with 1000 mg/kg bw/day and in females treated with 100 mg/kg bw/day. This observation is considered to be a result of local irritation following inhalation of microdroplets of the test compound as the gavage catheter was withdrawn. The absence of the finding at 300 mg/kg bw/day is further evidence that this is a procedure-related finding and as such, it is of limited toxicological significance.

Histopathological findings: neoplastic:

not specified

Details on results:

Necropsy and Gross Pathological Findings:
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 at necropsy. At microscopic examination, the treated animals were shown to have congestion in the lungs. Although there was an increased incidence/severity of congestion in the lungs the intergroup differences were considered to be within the normal range of background alterations that are seen in untreated animals of this age and strain and therefore was considered an incidental finding, not related to the treatments.

Histopathology:
The following treatment related microscopic abnormalities were detected:

Kidneys:increased incidence and severity of hyaline droplets and granular casts (consistent with alpha-2u-globulin nephropathy) were evident in males from all treatment groups. Although the findings of granular casts in the male kidney may be considered to represent an adverse effect of the test item, immunohistochemical staining demonstrated that these changes were correlated to accumulation of alpha 2u-globulin and formation of hyaline droplets, which is unique to the male rat. This finding is commonly observed in male rats following treatment with some hydrocarbons and is not predictive of any adverse effects in humans.

Granular casts may be considered to represent an adverse effect of the test item, however immunohistochemical staining demonstrated that these changes within the kidneys was correlated to the same condition as hyaline droplets therefore the kidney changes were considered to be of limited toxicological significance.

Stomach: Acanthosis, hyperkeratosis, ulceration and submucosal inflammation was evident in the forestomach of animals of either sex treated with 1000 and 300 mg/kg bw/day. The stomach changes identified in 1000 and 300 mg/kg bw/day animals were considered to be a result of local irritation of the test item rather than a true effect of systemic toxicity.

Lungs:increased incidence and severity of alveolar macrophages were evident in animals of either sex treated with 1000 mg/kg bw/day and in females treated with 100 mg/kg bw/day. This observation is considered to be a result of local irritation following inhalation of microdroplets of the test compound as the gavage catheter was withdrawn. The absence of the finding at 300 mg/kg bw/day is further evidence that this is a procedure-related finding and as such, it is of limited toxicological significance.

Organ Weights:
Males treated 1000 mg/kg bw/day and females from all treatment groups showed a statistically significant increase in liver weight both absolute and relative to terminal body weight (p<0.01; males, p<0.05; females). Although a number of the individual values were outside of the normal background range, no true dose related response for females was evident. In the absence of any associated histopathological correlates, the intergroup differences were therefore considered of no toxicological importance.

Males treated with 1000 mg/kg bw/day showed a statistically significant reduction in epididymides weight both absolute and relative to terminal body weight (p<0.05). All of the individual values were within the normal backgroundrange and in the absence of any associated histopathological correlates the intergroup difference was considered to be of no toxicological significance.

Clinical Biochemistry:
There were no toxicologically significant effects detected in the blood chemical parameters examined.

Males treated with 1000 mg/kg bw/day showed a statistically significant increase in alanine aminotransferase and females from this treatment group showed a statistically significant increase in urea (p<0.01). The majority of individual values
for these parameters were within the normal background ranges and in the absence of any associated histopathological correlates the intergroup differences were considered to be of no toxicological significance.

Males from all treatment groups showed statistically significant reductions in sodium concentration (p<0.05), potassium concentration (p<0.05) and chloride concentration (p<0.05) and a statistically significant increase in inorganic phosphorus
(p<0.05). Males treated with 100 mg/kg bw/day also showed a statistically significant increase in bilirubin (p<0.01). Females treated with 1000 and 300 mg/kg bw/day showed a statistically significantincrease in glucose (p<0.05). The majority of individual values for these parameters were within the normal background ranges and in the absence of true doserelated responses the intergroup differences were considered to be of no toxicological significance.

Hematology:
Males from all treatment groups showed a statistically significant increase in mean corpuscular hemoglobin concentration (p<0.05). The majority of individual values were within the normal background range and in the absence of a true dose related responsethe intergroup differences were considered to be of no toxicological significance.

Functional Performance Tests:
There were no toxicologically significant changes in functional performance at dosages of 100, 300 or 1000 mg/kg bw/day.

Males from all treatment groups and females treated with 1000 and 300 mg/kg bw/day showed a statistically significant reduction in overall activity when compared with controls (p<0.05-0.01). Females treated with 1000 and 300 mg/kg bw/day also showed a statistically significant reduction in activity during the final 20% period of observation (p<0.05). In the absence of a true dose related response and/orany supporting clinical observations to suggest an effect of neurotoxicity, the intergroup differences were considered to be of no toxicological significance.

Males treated with 1000 mg/kg bw/day showed a statistically significant reduction (p<0.05) in the first fore limb and hind limb grip strength test and a statistically significant increase in the final hind limb grip strength test. In the absence of a consistent effect or any supporting clinical observations to suggest an effect of neurotoxicity, the intergroup differences were considered to be of no toxicological significance.

Key result

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.

Key result

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.

Key result

Critical effects observed:

no

Table 1. Group Mean Functional Performance Test Values - Males
Group (sex)		Test 1 Forelimb (g)	Test 1 Hindlimb (g)	Test 2 Forelimb (g)	Test 2 Hindlimb (g)	Test 3 Forelimb (g)	Test 3 Hindlimb (g)	Overall Activity	Overall Mobile	Last 20% Activity	Last 20% Mobile
1 (M)	Mean	1234.3	432.8	1157.1	406.0	1022.3	347.1	1093.8	1.8	137.7	0.2n
	S.D.	240.7	107.7	285.2	62.1	162.9	99.5	300.2	1.9	118.4	0.4
	N	10	10	10	10	10	10	10	10	10	10
2 (M)	Mean	1067.0	385.9	1093.5	377.3	1060.0	425.2	797.6**	0.4	49.2	0.0n
	S.D.	276.5	116.5	300.8	98.2	243.1	95.1	225.2	0.5	62.4	0.0
	N	10	10	10	10	10	10	10	10	10	10
3 (M)	Mean	1217.5	425.3	1084.0	422.4	1069.8	405.6	862.5**	1.1	87.7	0.0n
	S.D.	237.5	104.6	276.5	134.2	195.5	74.4	149.0	1.6	81.4	0.0
	N	10	10	10	10	10	10	10	10	10	10
4 (M)	Mean	943.8*	325.8*	1101.5	446.0	990.0	442.2*	820.7*	1.0	72.4	0.0n
	S.D.	186.3	84.6	198.7	128.5	178.7	127.1	248.2	2.2	58.5	0.0
	N	10	10	10	10	10	10	10	10	10	10

General Footnote: Unit = Time (seconds) for Motor Activity Assessments

Dose Levels: Group 1 - 0(Control); Group 2 - 100 mg/kg bw/day; Group 3 - 300 mg/kg bw/day; Group 4 - 1000 mg/kg bw/day

 

Table 2. Group Mean Functional Performance Test Values - Females
Group (sex)		Test 1 Forelimb (g)	Test 1 Hindlimb (g)	Test 2 Forelimb (g)	Test 2 Hindlimb (g)	Test 3 Forelimb (g)	Test 3 Hindlimb (g)	Overall Activity	Overall Mobile	Last 20% Activity	Last 20% Mobile
1 (F)	Mean	966.3	309.4	866.0	293.4	904.5	291.9	803.1	5.3	122.1	1.2
	S.D.	255.2	73.7	185.3	97.7	174.1	66.4	266.0	9.9	113.4	2.6
	N	10	10	10	10	10	10	10	10	10	10
2 (F)	Mean	992.2	238.8	1004.0	232.4	963.2	308.4	724.2	6.2	89.7	2.0
	S.D.	224.9	68.5	191.6	95.7	174.6	124.4	619.7	17.9	168.6	6.3
	N	10	10	10	10	10	10	10	10	10	10
3 (F)	Mean	985.6	242.3	1011.2	288.3	857.3	255.6	498.4*	1.3	33.3*	0.1
	S.D.	147.0	93.5	182.1	74.4	183.1	115.0	256.0	2.1	50.5	0.3
	N	10	10	10	10	10	10	10	10	10	10
4 (F)	Mean	1024.5	302.9	936.2	296.9	1040.8	288.9	544.5*	1.8	22.0*	0.2
	S.D.	175.2	108.5	258.3	109.4	293.8	89.6	201.7	1.3	33.7	0.6
	N	10	10	10	10	10	10	10	10	10	10

General Footnote: Unit = Time (seconds) for Motor Activity Assessments

Dose Levels: Group 1 - 0(Control); Group 2 - 100 mg/kg bw/day; Group 3 - 300 mg/kg bw/day; Group 4 - 1000 mg/kg bw/day

Table 3. Group Mean Hematological Values - Males
Group (sex)		MCHC (g/dL)
Group 1 (0 – Control)	Mean	33.83
	S.D.	0.18
	N	10
Group 2 (100 mg/Kg bw/day)	Mean	34.38*
	S.D.	0.46
	N	10
Group 3 (300 mg/Kg bw/day)	Mean	34.02*
	S.D.	0.28
	N	10
Group 4 (1000 mg/Kg bw/day)	Mean	34.23*
	S.D.	0.4
	N	10

Table 4. Group Mean Blood Chemical Values - Males
Group (sex)		Na^+ mmol/L	K^+ mmol/L	Cl^- mmol/L	P mmol/L	ALAT IU/L	Bili mg/dL
Group 1 (0 – Control)	Mean	150.7	4.713	102.6	1.70	55.5	0.090
	S.D.	2.1	0.291	1.4	0.19	10.7	0.012
	N	10	10	10	10	10	10
Group 2 (100 mg/Kg bw/day)	Mean	147.6*	4.410*	100.6*	1.95*	58.6	0.165**
	S.D.	1.7	0.264	1.3	0.15	10.3	0.063
	N	10	10	10	10	10	10
Group 3 (300 mg/Kg bw/day)	Mean	148.6*	4.489*	101.4*	1.91*	92.7	0.099
	S.D.	3.9	0.373	1.9	0.17	111.6	0.014
	N	10	10	10	10	10	10
Group 4 (1000 mg/Kg bw/day)	Mean	148.7*	4.429*	101.4*	1.91*	66.7*	0.092
	S.D.	1.5	0.196	1.3	0.24	8.9	0.008
	N	10	10	10	10	10	10

 

Table 5. Group Mean Blood Chemical Values - Females
Group (sex)		Urea (mg/dL)	Glucose (mg/dL)
Group 1 (0 – Control)	Mean	38.1	123.7
	S.D.	5.7	18.4
	N	10	10
Group 2 (100 mg/Kg bw/day)	Mean	40.9	121.9
	S.D.	7.3	15.8
	N	10	10
Group 3 (300 mg/Kg bw/day)	Mean	39.6	145.4*
	S.D.	6.1	17.2
	N	10	10
Group 4 (1000 mg/Kg bw/day)	Mean	47.3**	137.0*
	S.D.	4.7	17.0
	N	10	10

Table 6. Group Mean Organ Weights with Corresponding Relative (% of Body Weight) Organ Weights
		Males				Females
		0 Control	100 mg/Kg bw/day	300 mg/Kg bw/day	1000 mg/Kg bw/day	0 Control	100 mg/Kg bw/day	300 mg/Kg bw/day	1000 mg/Kg bw/day
Epididymides	Mean (g)	1.73769	1.66163	1.64448	1.60713*
	S.D.	0.16852	0.16045	0.10688	0.12939
	N	10	10	10	10
	Mean (%)	0.430	0.395	0.392	0.387*
	S.D.	0.037	0.067	0.027	0.035
	N	10	10	10	10
Liver	Mean (g)	12.6953	13.8934	14.0717	15.1921**	8.37888	8.68144*	8.55061*	8.88699*
	S.D.	2.20822	2.07704	0.84560	1.33677	0.65274	0.87496	0.77236	0.95142
	N	10	10	10	10	10	10	10	10
	Mean (%)	3.120	3.246	3.347	3.652**	3.345	3.605*	3.468*	3.591*
	S.D.	0.314	0.244	0.102	0.298	0.286	0.080	0.224	0.214
	N	10	10	10	10	10	10	10	10

 

Table 7. Summary Incidence of Necropsy Findings - Males
	Males
	0 Control	100 mg/Kg bw/day	300 mg/Kg bw/day	1000 mg/Kg bw/day
Number of animals examined	10	10	10	10
Lungs (With Bronchi)
Submitted	(10)	(10)	(10)	(10)
No Visible Lesions	10	10	9	10
Reddened	0	0	1	0

 

Table 8. Summary Incidence of Necropsy Findings - Females
	Females
	0 Control	100 mg/Kg bw/day	300 mg/Kg bw/day	1000 mg/Kg bw/day
Number of animals examined	10	10	10	10
Heart
Submitted	(10)	(10)	(10)	(10)
No Visible Lesions	10	10	10	9
Mass	0	0	0	1
Lungs (With Bronchi)
Submitted	(10)	(10)	(10)	(10)
No Visible Lesions	9	9	8	5
Discolouration; Red	1	0	2	4
Reddened	0	1	0	0
Mass	0	0	0	1
Vagina
Submitted	(10)	(10)	(10)	(10)
No Visible Lesions	10	10	10	9
Fluid Filled	0	0	0	1

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: This study is classified as reliable with restrictions because there was no statement to indicate that it was conducted according to GLP standards, but it was generally conducted according to OECD 412 guidelines.

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in Section 13.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Deviations:

no

GLP compliance:

not specified

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

other: unchanged (no vehicle)

Duration of treatment / exposure:

6 hours a day

Frequency of treatment:

5 days a week for 2 weeks

Remarks:

Doses / Concentrations:
300, 1000, 3000, or 8000 ppm
Basis:
nominal conc.

No. of animals per sex per dose:

Ten rats per sex per dose

Control animals:

yes, sham-exposed

Dose descriptor:

LOAEC

Effect level:

8 000 ppm

Basis for effect level:

other: Based on decreased female body weight

Dose descriptor:

NOAEC

Effect level:

3 000 ppm

Critical effects observed:

not specified

Conclusions:

The study did not identify a NOAEC; a LOAEC of 8000 ppm could be selected based on body weight effects in females, resulting in a NOAEC of 3000 ppm.

Executive summary:

In a subacute inhalation toxicity study, Neodene-6 alpha olefin was administered to 10 Fischer 344 rats/sex/concentration by dynamic whole body exposure at concentrations of 0, 300, 1000, 3000 or 8000 ppm for 6 hours per day, 5 days/week for 2 weeks for a total of 11 exposures.

 

Females exposed to 8000 ppm exhibited slight (<10% change from controls), but significant, decreased body weight. There was a significant, dose-dependent decrease in alkaline phosphatase activity in all male treatment groups, but no changes in liver weight or histopathology. Alkaline phosphatase activity was decreased in high-dose females, but not significantly. absolute and relative liver and kidney weights were increased in 8000 -ppm females, likely due to the weight loss in this group as there were no histopathological findings in these organs. Neuromuscular coordination (Rotarod test) was significantly reduced in the 300 -, 3000 -, and 8000 -ppm females compared to the controls after 5 days, but not after 12 days; the authors interpreted the initial findings to stress and not an effect of the test compound. No neuromuscular deficits were noted in male rats. The LOAEC is 8000 ppm based on reduced body weight in females. The NOAEC is 3000 ppm.

 

This study received a Klimisch score of 2 and is classified as reliable with restrictions because there was no statement to indicate that the study was conducted according to GLP standards, but it was generally conducted according to OECD 412 guidelines.

This study was selected as a supportive study because of the short exposure period (i.e., 11 days vs. 28 days as recommended by OECD guidelines).

Reference 2

Endpoint:

sub-chronic toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1982-12-21 to 1983-04-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study is classified as reliable without restrictions because it was conducted according to OECD 413 guidelines and was GLP compliant.

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in Section 13.

Reason / purpose for cross-reference:

read-across: supporting information

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

Only a few major organs were examined histologically.

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Sprague-Dawley, Inc.
- Fasting period before study: No
- Housing: Individually
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: 19 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24±2°C
- Humidity (%): 40 to 70%
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

IN-LIFE DATES: From: 1983-01-07 To: 1983-04-22

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

other: unchanged (no vehicle)

Remarks on MMAD:

MMAD / GSD: None

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:7700 Litre King-Lar stainless steel/glass chambers housed in inhalation suites
- Method of holding animals in test chamber: None
- Source and rate of air: Main air supply line at an unspecified rate
- System of generating particulates/aerosols: A head space sweep technique using dry nitrogen through a mass flow controller into a stainless steel container supplied with a continuous feed of test material, then introduced into the main air supply line with a static mixing device, then fed into the exposure chamber
- Temperature, humidity, pressure in air chamber: 22 to 26 degrees Celsius and 30 to 63% humidity
- Air flow rate: 533 cubic meters to 578 cubic meters
- Air change rate: 1.3 to 1.6 cubic meters per minute

TEST ATMOSPHERE
- Brief description of analytical method used: The concentration of hex-1-ene was determined by gas chromatography using an 80/100 mesh Carbopak® column with 0.19% picric acid and flame ionization detection. Vapour concentration was determined about every ten minutes during exposure. Samples were taken near the centre of the chamber where the animals were positioned in the exposure chamber. Three Varian Vista Series 4600 Gas Chromatographs, calibrations performed daily, with one CDS 401 Chromatography Data System was used for the concentration analyses.
- Samples taken from breathing zone: Yes

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analytical concentrations for the low, middle, and high exposures ranged from 297 to 300 parts per million, 990 to 1002 parts per million, and 2958 to 3000 parts per million, respectively. The analytical concentrations were slightly (<10%) lower than the calculated nominal concentrations.

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

6 hours a day, 5 days a week

Remarks:

Doses / Concentrations:
300, 1000, or 3000 parts per million
Basis:
analytical conc.

No. of animals per sex per dose:

Forty animals/sex/dose; ten animals/sex/dose for neuromuscular testing, ten animals/sex/dose for interim sacrifice at 7 weeks, and twenty animals/sex/dose for terminal sacrifice

Control animals:

yes, sham-exposed

Details on study design:

- Dose selection rationale: Based on an 11-day study (WRC-TIR-733)

Positive control:

none

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes, mortality and clinical signs
- Time schedule: Before and after exposure and once on days with no exposure

DETAILED CLINICAL OBSERVATIONS: No data reported.

BODY WEIGHT: Yes
- Time schedule for examinations: Pretreatment and weekly during exposure

FOOD CONSUMPTION: No

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Pretest, week 7 and week 13
- Anaesthetic used for blood collection: No data reported
- Animals fasted: Yes
- How many animals: Ten animals per sex and dose at study initiation, ten animals per sex and dose at 7 weeks, and twenty animals per sex and dose at week 13
- Parameters checked in table 1 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Pretest, week 7, and week 13
- Animals fasted: Yes
- How many animals: Ten animals per sex and dose at study initiation, ten animals per sex and dose at 7 weeks, and twenty animals per sex and dose at week 13
- Parameters checked in table 2 were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: Weeks 7 and 13
- Metabolism cages used for collection of urine: No data reported.
- Animals fasted: Yes
- Parameters checked in table 3 were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Pretest and weekly during exposure
- Dose groups that were examined: All groups
- Battery of functions tested: Balance and neuromuscular coordination

OTHER: Sperm counts

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes (see table 4)

Other examinations:

The lungs, liver, kidney, brain, heart, spleen, and right testicle or uterus were weighed. Sperm counts, measured with a hemocytometer and phase-contrast microscope, were performed using the left testicle of each male rat after detunication, weighing, and homogenization.

Statistics:

Body weight differences were analyzed by analysis of covariance using initial weights of individual rats on day -2 as a covariate with the least squares difference test. Organ weight, clinical chemistry, haematology, urinalysis, and organ to body weight ratio data were analyzed by Dunnett’s t-test on the ranked data. Neuromuscular coordination data were adjusted by summing the time for the best 3 of 4 trials for each rat and analyzed by analysis of variance or analysis of covariance. All statistics were measured with p<0.05.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY: No deaths occurred. There were no clinical signs related to treatment.

BODY WEIGHT AND WEIGHT GAIN: Decreased body weight was observed in 3000-ppm females (statistically significant) and males (statistically significant only sporadically). The reported 4% reduction in body weight, while statistically significant, is of limited toxicological significance. (Table 5)

HAEMATOLOGY: There were no treatment-related findings at interim sacrifice. At terminal sacrifice, 1000-ppm females and 3000-ppm males and females had higher haematocrit and red blood cells, and in 1000-ppm and 3000-ppm females mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were lower. Although these findings appear treatment-related, they were not associated with any histopathology findings, results are not dose related, and changes were within 5% of the control. (Table 6)

CLINICAL CHEMISTRY: There was an increase in phosphorus in males at all treatment concentrations, which may be due to a low level in the control. There was a consistent increase in phosphorus in 1000-ppm and 3000-ppm females at interim and terminal sacrifice (Table 7). The toxicological significance of this finding is doubtful.

URINALYSIS: There were no treatment-related changes.

NEUROBEHAVIOUR: There were no treatment-related effects.

ORGAN WEIGHTS: There was a slight (5% or less) but significant decrease in absolute and relative testes weight in 3000-ppm males that was not associated with any change in sperm parameters or histopathology. This decrease, while statistically significant, is of limited toxicological relevance. Other changes in organ weights were related to the reduced body weight.

GROSS PATHOLOGY: There were no treatment-related findings.

HISTOPATHOLOGY: NON-NEOPLASTIC: There were no treatment-related findings.

HISTOPATHOLOGY: NEOPLASTIC (if applicable): There were no treatment-related findings.

OTHER FINDINGS: There were no treatment-related changes in sperm counts.

Dose descriptor:

NOAEC

Effect level:

3 000 ppm

Sex:

male/female

Basis for effect level:

other: Equivalent to 10,326 mg/m3; no toxicologically relevant findings at the highest concentration tested.

Critical effects observed:

not specified

Table 5. Body weight data

 

Body weight data expressed in grams (mean ± s.d.)
	Males - ppm
	0	300	1000	3000
Day 0	161.28 ± 2.61 n=40	159.84 ± 2.58 n=40	159.72 ± 2.93 n=40	161.58 ± 2.37 n=40
Day 33	250.1 ± 3.9 n=40	252.3 ± 3.1 n=40	246.2 ± 4.4 n=40	245.0 ± 3.2 n=40
Day 61	305.2 ± 4.3 n=30	305.2 ± 3.8 n=30	302.3 ± 5.3 n=30	295.1 ± 3.7 n=30
Day 89	338.9 ± 4.2 n=30	340.8 ± 4.0 n=30	339.7 ± 5.2 n=30	329.0 ± 3.6 n=30
	Females – ppm
	0	300	1000	3000
Day 0	126.04 ± 1.29 n=40	125.30 ± 1.28 n=40	125.69 ± 1.28 n=40	125.94 ± 1.31 n=40
Day 33	164.44 ± 1.90 n=40	162.93 ± 1.36 n=40	163.82 ± 1.47 n=40	160.18 ± 1.48 n=40
Day 61	183.80 ± 2.47 n=30	181.83 ± 1.61 n=30	181.23 ± 1.73 n=30	175.41 ± 1.75 * n=30
Day 89	194.87 ± 2.62 n=30	194.62 ± 1.68 n=30	192.93 ± 2.02 n=30	187.08 ± 1.85 * n=30

*Statistically significant at p<0.05.

Table 6. Selected Hematological Data

Hematological Data (mean ± s.d.)
	Males - ppm
	0	300	1000	3000
Haematocrit (%) (week 13)	44.4 ± 0.5 n=20	43.7 ± 0.7 n=20	45.0 ± 0.1 n=20	45.3 ± 0.3* n=20
RBC (10^6/ mm^3) (week 13)	6.58 ± 0.07 n=20	6.49 ± 0.12 n=20	6.65 ± 0.03 n=20	6.71 ± 0.05 * n=20
	Females – ppm
	0	300	1000	3000
Haematocrit (%) (week 13)	40.9 ± 0.3 n=20	42.5 ± 0.7 n=20	43.1 ± 0.8 * n=20	42.6 ± 0.05 * n=20
RBC (10^6/ mm^3) (week 13)	5.78 ± 0.05 n=20	6.01 ± 0.10 n=20	6.09 ± 0.11 * n=20	6.04 ± 0.07 * n=20

*Statistically significant at p<0.05

Table 7. Phosphorus levels in the blood

Clinical Chemistry Data (mean ± s.d.)
	Males - ppm
	0	300	1000	3000
Phosphorus mg/dL (week 7)	6.98 ± 0.23 n=10	7.21 ± 0.26 n=10	8.18 ± 0.29 * n=10	7.52 ± 0.23 n=10
Phosphorus mg/dL (week 13)	6.42 ± 0.15 n=20	7.27 ± 0.20 * n=20	7.22 ± 0.14 * n=20	7.72 ± 0.34 * n=20
	Females – ppm
	0	300	1000	3000
Phosphorus mg/dL (week 7)	6.28 ± 0.57 n=10	6.39 ± 0.42 n=10	7.59 ± 0.37 * n=10	7.78 ± 0.27 * n=10
Phosphorus mg/dL (week 13)	5.64 ± 0.19 n=20	6.07 ± 0.24 n=20	6.46 ± 0.24 * n=20	6.85 ± 0.15 * n=20

 *Statistically significant at p<0.05.

Conclusions:

The NOAEC for this study was 3000 ppm (10,326 mg/m3) based on a lack of toxicologically relevant findings at the highest concentration tested.

Executive summary:

In a 90-day inhalation toxicity study, Neodene 6 alpha olefin was administered to forty Fischer 344 rats/sex/concentration by dynamic whole body exposure at concentrations of 0, 300, 1000, or 3000 parts per million (corresponding to 0; 1033; 3442; or 10, 326 mg/m³) for 6 hours a day, 5 days a week, for 13 weeks. Ten of the animals/sex/concentration were used for neuromuscular testing, ten of the animals/sex/concentration were sacrificed after 7 weeks of exposure, and twenty animals/sex/concentration were sacrificed after 13 weeks of exposure.

Subchronic inhalation of Neodene 6 alpha olefin for 13 weeks did not produce any adverse respiratory, neuromuscular, or testicular effects in rats. Decreased body weight was observed in 3000 -ppm females (statistically significant) and males (statistically significant only sporadically). Decreased absolute liver and kidney weights were observed in 3000-ppm females; however, these findings were considered secondary to reduced body weight in the absence of histopathological findings in these organs. There were statistically significant differences in haematology and clinical chemistry values, but the changes were slight (generally within 5% of the control), were not dose related, and/or not associated with any histopathology findings.Increased phosphorus levels were reported in males at all treatment levels and females exposed to 1000 and 3000 ppm hex-1-ene. The toxicological significance of these findings is doubtful. The NOAEC is 3000 parts per million (10,326 mg/m³) based on a lack of toxicologically relevant findings at the highest concentration tested.

This study received a Klimisch score of 1 and is classified as reliable without restrictions because the study was conducted according to OECD 413 guidelines and was GLP compliant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: This study is classified as reliable not assignable because this study only examined one dose level, and systemic toxicity was not evaluated.

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in Section 13.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)

Deviations:

yes

GLP compliance:

no

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

not specified

Details on test animals or test system and environmental conditions:

No data reported with regard to test animals or environmental conditions.

IN-LIFE DATES: From: not reported To: 28 days later

Type of coverage:

not specified

Vehicle:

unchanged (no vehicle)

Details on exposure:

No data was reported with regard to area of exposure, percentage of coverage, type of wrap (if used), or time intervals for shavings/clippings.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.2 millilitres
- Constant volume or concentration used: Yes

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

No data reported.

Frequency of treatment:

5 days a week for 4 weeks for a total of 20 applications

Remarks:

Doses / Concentrations:
0.2 millilitres
Basis:
no data

No. of animals per sex per dose:

Six animals per dose (3 intact and 3 abraded)

Control animals:

no

Details on study design:

No data reported.

Positive control:

None

Observations and examinations performed and frequency:

There were no data reported regarding whether cageside observations or detailed clinical observations were performed. Additionally, there were no data reported with regard to the methods used (if any) for following examinations: body weight, food consumption, food efficiency, water consumption, ophthalmoscopic, haematology, clinical chemistry, urinalysis, and neurobehavioural.
DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: 24 hours after each application

Sacrifice and pathology:

Methods for gross pathology or histopathology examinations were not reported.

Statistics:

No data reported.

Clinical signs:

not specified

Dermal irritation:

effects observed, treatment-related

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Details on results:

OTHER FINDINGS: The study authors reported that animals exposed to 1-octene exhibited questionable hyperaemia, exfoliation, and scab formation.

Dose descriptor:

NOAEL

Sex:

not specified

Basis for effect level:

other: Report only specified dermal irritation and results indicated questionable irritation.

Remarks on result:

not determinable

Remarks:

no NOAEL identified

Critical effects observed:

not specified

The average results for the hyperaemia, exfoliation and scab formation were approximately 2, which fell into the study authors questionable grade. There was no difference in reaction between the abraded and intact animals.

Conclusions:

Using a graded scale of 1 to 6 (none to severe), treatment with 1-octene for 28 days (20 treatments) caused an average score of approximately 2 (considered questionable) for hyperaemia, exfoliation, and scab formation.

Executive summary:

In a 28-day dermal toxicity study, alpha olefin 8 was applied to the shaved skin of six New Zealand white rabbits (3 intact and 3 abraded) at a dose level of 0.2 millilitres for 5 days/week during a 28-day period.

 

The skin was ranked using a graded system by the study authors ranging from 1 (none) to 6 (severe). The average score for hyperaemia, exfoliation, and scab formation was approximately 2 (considered questionable). There were no other toxicity results reported. Therefore, no LOAEL or NOAEL were identified.

 

This study received a Klimisch score of 4 and is classified as not assignable because this study only examined one dose level, and systemic toxicity was not evaluted.

This study was selected as a supporting study because there was only was dose level examined and systemic toxicity was not evaluated; however, the study contains relevant and useful information, allowing this study to qualify as a supporting study.

Reference 2

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Read-across between the target substance Oct-2-ene (EC 203-894-2 / CAS 111-67-1) and source substances Octene (EC 246-920-8 / CAS 25377-83-7) and Oct-1-ene (EC 203-893-7 / CAS 111-66-0 / alpha-C8) is based upon the similarity of the chemical structures and their respective physico-chemical properties. The ECHA Read-Across Assessment Framework (RAAF) states that substances with qualitatively similar properties can form the basis of read-across in circumstances where the source and target substances share such similar characteristics.

Octene, Oct-1-ene and Oct-2-ene are linear olefins, each with a carbon chain length of C8. Structurally, the difference between source and target substances is the position of the carbon-carbon double bond. For Oct-1-ene the double bond is at the terminal C1 position (hence, an alpha-olefin), whereas for Oct-2-ene the double bond is at the non-terminal C2 position (hence, an internal olefin). A comparison of the target and source substance properties shows that all substances would be expected to exhibit similar environmental fate, ecotoxicological and mammalian toxicological behaviours. The justification for read-across from source substances Octene and Oct-1-ene to target substance Oct-2-ene is detailed in section 13 (Document name: “Oct-2-ene Read Across Document HOPA”).

Further, Oct-2-ene and Oct-1-ene both fit within the boundaries of the chemical category of higher olefins. Studies conducted by the HOPA consortium on a large range of higher olefin category members (including Oct-1-ene) demonstrated sufficiently similar physico-chemical, environmental fate and toxicological properties to substantiate the basis for read-across. Therefore Oct-2-ene is expected to behave similarly. Justification for inclusion of Oct-2-ene within the boundaries of the higher olefins category, and the relevance of each category member as an analogue substance to Oct-2-ene, is provided in Section 13 (Document name: “HOPA Higher Olefins CJD with Category Matrix Report [rev 1 Sept 2016]”).

Reason / purpose for cross-reference:

read-across: supporting information

Reason / purpose for cross-reference:

read-across source

GLP compliance:

no

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

No data reported.

Frequency of treatment:

5 days a week for 4 weeks for a total of 20 applications

Remarks:

Doses / Concentrations:
0.2 millilitres
Basis:
no data

Statistics:

No data reported.

Clinical signs:

not specified

Dermal irritation:

effects observed, treatment-related

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Details on results:

OTHER FINDINGS: The study authors reported that animals exposed to 1-octene exhibited questionable hyperaemia, exfoliation, and scab formation.

Dose descriptor:

NOAEL

Sex:

not specified

Basis for effect level:

other: Report only specified dermal irritation and results indicated questionable irritation.

Remarks on result:

not determinable

Remarks:

no NOAEL identified

Critical effects observed:

not specified

The average results for the hyperaemia, exfoliation and scab formation were approximately 2, which fell into the study authors questionable grade. There was no difference in reaction between the abraded and intact animals.

Conclusions:

Using a graded scale of 1 to 6 (none to severe), treatment with 1-octene for 28 days (20 treatments) caused an average score of approximately 2 (considered questionable) for hyperaemia, exfoliation, and scab formation.

Executive summary:

In a 28-day dermal toxicity study, the read-across source alpha olefin 8 was applied to the shaved skin of six New Zealand white rabbits (3 intact and 3 abraded) at a dose level of 0.2 millilitres for 5 days/week during a 28-day period.

The skin was ranked using a graded system by the study authors ranging from 1 (none) to 6 (severe). The average score for hyperaemia, exfoliation, and scab formation was approximately 2 (considered questionable). There were no other toxicity results reported. Therefore, no LOAEL or NOAEL were identified.

Read-across of this result to Oct-2 -ene is claimed as valid basd on the justifications provided for both analogue and category approaches.

 

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Repeated dose toxicity studies in rats have been conducted for 14 members of the higher olefin category, covering C6 to C20-24. The majority of substances in this category appear to possess limited potential to cause systemic toxicity following repeated oral exposure. While available sub-chronic toxicity data for hex-1-ene, tetradec-1-ene, and octadec-1-ene suggest some potential for systemic toxicity in rats, the effects observed were either minor or adaptive in nature and occurred at dose levels greater than 100 mg/Kg bw/day. Repeated oral exposure to the other substances in the category did not result in any systemic toxicity in rats at doses as high as 1000 mg/Kg bw/day. Therefore, based on the weight of evidence, substances in this category do not warrant classification for repeat dose oral toxicity under the current CLP regulation.   

Repeat dose inhalation toxicity data available from studies conducted using hex-1-ene indicate a NOAEC of 3000 ppm (10,326 mg/m³) in rats. Therefore, substances in this category do not warrant classification for repeat dose inhalation toxicity under the current CLP regulation.  

Limited data are available on the repeated dose dermal toxicity potential of higher olefin category substances. The physicochemical and toxicological properties of higher olefins (linear alpha; isomerised; alpha, internal, linear and branched – single and multiple carbon numbers) do not suggest potential to readily penetrate the skin or to be absorbed at a significant rate through the skin. Additionally, results from acute oral and dermal toxicity studies in animals affirm that these substances are not toxic by either route. Repeated oral administration of higher olefins for up to 90 days has not been associated with significant or long-lasting effects. Based on the weight of evidence provided by the physicochemical data, and acute as well as repeat dose toxicity data, substances in this category do not warrant classification for repeat dose dermal toxicity under the current CLP regulation.

Read-across of these results to Oct-2 -ene is claimed as valid based on the justifications provided for both analogue and category approaches