Registration Dossier
Registration Dossier
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EC number: 203-894-2 | CAS number: 111-67-1
Endpoint summary
Administrative data
Key value for chemical safety assessment
Effects on fertility
Description of key information
Guideline (OECD 421 and 422) reproductive toxicity screening studies have been conducted in rats, for nine members of the higher olefin category, covering C6 to C18. These investigations have all used oral (gavage) exposure. Oral toxicity data is supported by results from a 90-day sub-chronic inhalation study in rats with a C8 olefin.
Except for one study with Nonene, branched, where the NOAEL for reproductive toxicity was considered to be 300 mg/Kg bw/day based on reduced post-natal offspring viability, offspring body weight gain, and litter size at 1000 mg/Kg bw/day, no effects on reproduction were observed in any other study at oral dose levels up to 1000 mg/Kg bw/day.
Inhalation exposure up to 10,326 mg/m3 did not reveal any effects on reproductive organs.
Read-across of these results to Oct-2 -ene is claimed as valid based on the justifications provided for both analogue and category approaches.
Link to relevant study records
- Endpoint:
- screening for reproductive / developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key 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:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Oct-1-ene
- Cs No.: 111-66-0
- Purity test date: >98%
- Lot/batch No.: 719238
- Expiration date of the lot/batch: 29 April 2014
- Label: Alpha Olefin C8 Lot 719238
- Data received: 29 April 2013
- Storage condition of test material: room temperature in the dark under nitrogen
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 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 - Details on mating procedure:
- - M/F ratio per cage: 1/1 (pairing period)
- Length of cohabitation: minimum 14 days
- Proof of pregnancy: The presence of sperm within the vaginal smear and/or vaginal plug in situ was taken as positive evidence of mating (Day 0 of gestation)
- After successful mating each pregnant female was caged: males were returned to their original holding cages (unless required for additional pairing). Mated females were housed individually during the period of gestation and lactation. - 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
- Details on study schedule:
- 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. - 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
- Parental animals: Observations and examinations:
- 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.
DOOD CONSUMPTION:Yes
Pre-pairing period and after (male): weekly
Females showing evidence of mating: post coitum Days 0-7, 7-14 and 14-20.
Females with live litters: Days 1 and 4 post partum.
FOOD EFFICIENCY:Yes
Males (throughout the study period-with the exception of the mating phase, and females during the pre-pairing phase
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Water intake was measured daily during the pre-pairing phase of the
BEHAVIORAL, FUNCTIONAL OBSERVATIONS and SENSORY REACTIVITIES were observed and recorded. - Litter observations:
- Number of offspring born
Number of offspring alive recorded daily and reported on Days 1 and 4 post partum
Sex of offspring on Days 1 and 4 post partum
Clinical condition of offspring from birth to Day 5 post partum
Individual offspring weights on Days 1 and 4 post partum (litter weights were calculated retrospectively from this data)
- Postmortem examinations (parental animals):
- 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.
- Postmortem examinations (offspring):
- All adult animals and offspring, including those dying during the study, were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.
- 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).
- Reproductive indices:
- Mating Performance and Fertility
i. Pre-coital Interval
ii. Fertility Indices (Mating Index and Pregnancy Index)
Gestation and Parturition Data
i. Gestation Length
ii. Parturition Index - Offspring viability indices:
- Parturition Index (%)
Litter Responses
i. Implantation Losses (%)
ii. Live Birth and Viability Indices
iii. Sex Ratio (% males) - 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 ingroup 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
- 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
- Reproductive function: oestrous cycle:
- not examined
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- Mating
There were no treatment-related effects on mating performance.
Fertility
No treatment-related effects on fertility were detected for treated animals, when compared to controls. One control female and one female treated with 100 mg/kg bw/day did not achieve pregnancy following evidence of mating. No histopathological correlates were evident in the reproductive organs for these animals and in the absence of any similar infertility effects detected at 1000 mg/kg bw/day the intergroup differences were considered not to be related to test item toxicity.
Gestation Length
There were no differences in gestation lengths. The distribution for treated females was comparable to controls. The gestation lengths were between 22 and 23½ days. - Key result
- Dose descriptor:
- NOEL
- Remarks:
- systemic toxicity
- 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
- Remarks:
- systemic toxicity
- 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
- Dose descriptor:
- NOEL
- Remarks:
- reproductive toxicity
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No reproductive effects at 1000 mg/kg/day
- Key result
- Critical effects observed:
- no
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No obvious clinical signs of toxicity were detected for offspring from treated females when compared to controls. The incidental clinicalsigns detected throughout the control and treated groups, consisting of small size, no milk in stomach, a physical injury, found dead or missing, were considered to be low incidence findings observed in offspring in studies of this type and were considered unrelated to test item toxicity.
- Mortality / viability:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Sexual maturation:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Histopathological findings:
- not examined
- Key result
- Dose descriptor:
- NOEL
- Generation:
- F1
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No Developmental effects seen at 1000 mg/Kg/day
- Key result
- Critical effects observed:
- no
- Key result
- Reproductive effects observed:
- no
- 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. The ‘No Observed Effect Level’ (NOEL) for reproductive toxicity was considered to be 1000 mg/kg bw/day.
- 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.
No significant differences were detected for corpora lutea, implantation counts, implantation losses, litter size or litter viability for treated animals when compared to controls. Statistical analysis of the data did not reveal any significant intergroup differences
In view of these results, the NOAEL for systemic toxicity was considered to be 1000 mg/Kg bw/day. The 'No Observed Effect Level' (NOEL) for reproductive toxicity was considered to be 1000 mg/Kg bw/day.
- Endpoint:
- screening for reproductive / developmental toxicity
- 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
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Oct-1-ene
- Cs No.: 111-66-0
- Purity test date: >98%
- Lot/batch No.: 719238
- Expiration date of the lot/batch: 29 April 2014
- Label: Alpha Olefin C8 Lot 719238
- Data received: 29 April 2013
- Storage condition of test material: room temperature in the dark under nitrogen
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
- Vehicle:
- arachis oil
- 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 ingroup 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
- 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
- Reproductive function: oestrous cycle:
- not examined
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- Mating
There were no treatment-related effects on mating performance.
Fertility
No treatment-related effects on fertility were detected for treated animals, when compared to controls. One control female and one female treated with 100 mg/kg bw/day did not achieve pregnancy following evidence of mating. No histopathological correlates were evident in the reproductive organs for these animals and in the absence of any similar infertility effects detected at 1000 mg/kg bw/day the intergroup differences were considered not to be related to test item toxicity.
Gestation Length
There were no differences in gestation lengths. The distribution for treated females was comparable to controls. The gestation lengths were between 22 and 23½ days. - Key result
- Dose descriptor:
- NOEL
- Remarks:
- systemic toxicity
- 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
- Remarks:
- systemic toxicity
- 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
- Dose descriptor:
- NOEL
- Remarks:
- reproductive toxicity
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No reproductive effects at 1000 mg/kg/day
- Key result
- Critical effects observed:
- no
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No obvious clinical signs of toxicity were detected for offspring from treated females when compared to controls. The incidental clinicalsigns detected throughout the control and treated groups, consisting of small size, no milk in stomach, a physical injury, found dead or missing, were considered to be low incidence findings observed in offspring in studies of this type and were considered unrelated to test item toxicity.
- Mortality / viability:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Sexual maturation:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Histopathological findings:
- not examined
- Key result
- Dose descriptor:
- NOEL
- Generation:
- F1
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No Developmental effects seen at 1000 mg/Kg/day
- Key result
- Critical effects observed:
- no
- Key result
- Reproductive effects observed:
- no
- Conclusions:
- An OECD 422 study was conducted using the read-across source 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. The ‘No Observed Effect Level’ (NOEL) for reproductive toxicity was considered to be 1000 mg/kg bw/day.
Read-across of this result to Oct-2 -ene is claimed as valid based on the justifications provided for both analogue and category approaches - 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 read across osurce 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.
No significant differences were detected for corpora lutea, implantation counts, implantation losses, litter size or litter viability for treated animals when compared to controls. Statistical analysis of the data did not reveal any significant intergroup differences
In view of these results, the NOAEL for systemic toxicity was considered to be 1000 mg/Kg bw/day. The 'No Observed Effect Level' (NOEL) for reproductive toxicity was considered to be 1000 mg/Kg bw/day.
Read-across of this result to Oct-2 -ene is claimed as valid based on the justifications provided for both analogue and category approaches
- Endpoint:
- screening for reproductive / developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 1993-12-03 to 1994-08-26
- 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 421 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 421 (Reproduction / Developmental Toxicity Screening Test)
- Deviations:
- no
- GLP compliance:
- yes
- Limit test:
- no
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): The test material was composed of equal amounts of Neodene 6 alpha olefin (component A), hexene-1 Gulftene 6 (component B), and alpha olefin C6 1-hexene (component C).
- Substance type: C6 alpha olefin
- Physical state: Clear, colourless liquid
- Analytical purity: Approximately 99%
- Lot/batch No.: 20202-45-1049 (component A), CBN0044 (component B), and 300-974 (component C)
- Expiration date of the lot/batch: December, 1994 (component A), others not provided
- Storage condition of test material: Under nitrogen at room temperature - Species:
- rat
- Strain:
- Sprague-Dawley
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Laboratories, Inc., Portage, Michigan
- Age at study initiation: (P) x males were 6 weeks old and females were 8 weeks old; F1 not treated
- Weight at study initiation: (P) Males: 195 to 242 grams; Females: 163 to 219 grams; F1 not treated
- Fasting period before study: No
- Housing: Individually except during mating
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: 11 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 23 °C
- Humidity (%): 35 to 65%
- Air changes (per hr): 10 to 12
- Photoperiod (hrs dark / hrs light): 12 hours dark/ 12 hours light
IN-LIFE DATES: From: 1994-02-11 To: 1994-05 - Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: After stirring the blended test material for 15 minutes, a specified amount of test material was added to a flask with half of the corn oil. The flasks were capped and inverted several times, then the remaining corn oil was added and the procedure repeated. The solution was then stirred for 15 minutes. Fresh preparations were made bi-weekly.
VEHICLE
- Concentration in vehicle: 0, 20, 100, or 200 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg
- Lot/batch no. (if required): APR0695A, Oct0494A, Jun2695A, and Dec2194A - Details on mating procedure:
- - M/F ratio per cage: 1 to 1
- Length of cohabitation: 15 days
- Proof of pregnancy: Vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After 10 days of unsuccessful pairing replacement of first male by another male with proven fertility for a maximum of 5 days.
- Further matings after two unsuccessful attempts: No
- After successful mating each pregnant female was caged (how): Individual boxes containing nesting materials
- Any other deviations from standard protocol: None - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Homogeneity of the dose formulation was checked from upper, middle, and lower layers of 20 and 200 mg/mL samples. The dose formulation was found to be homogeneous. The stability of the dose formulation was tested on 20 and 200 mg/mL samples stored in the fridge for 3, 8, or 15 days. The samples were found to be stable for at least 15 days. Verification of the 0, 20, 100, and 200 mg/mL dose formulations prepared during week 1, 3, 5, 7, and 9 were conducted. All dose formulations were found to be within 10% of the nominal concentration.
- Duration of treatment / exposure:
- Males: 44 days starting 28 days prior to mating
Females: 41 to 55 days starting 14 days prior to mating through lactation day 4 - Frequency of treatment:
- Daily
- Details on study schedule:
- F1 animals were not mated
- Dose / conc.:
- 100 mg/kg bw/day (actual dose received)
- Remarks:
- Basis:
actual ingested
5.0 mL/kg of dose formulations of 20 mg/mL were used - Dose / conc.:
- 500 mg/kg bw/day (actual dose received)
- Remarks:
- Basis:
actual ingested
5.0 mL/kg of dose formulations of 100 mg/mL were used - Dose / conc.:
- 1 000 mg/kg bw/day (actual dose received)
- Remarks:
- Basis:
actual ingested
5.0 mL/kg of dose formulations of 200 mg/mL were used - No. of animals per sex per dose:
- twelve animals per sex per dose
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: A non-reproductive dose range finding study
- Positive control:
- None
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: No
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once daily, mortality was checked twice daily
BODY WEIGHT: Yes
- Time schedule for examinations: Weekly in males; weekly in females prior to mating, then on gestational days 0, 7, 14, and 20 and lactation days 1 and 4
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
- Oestrous cyclicity (parental animals):
- Not examined
- Sperm parameters (parental animals):
- Parameters examined in P male parental generations: Testis weight and epididymis weight and histopathology
- Litter observations:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: No
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities
GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead. - Postmortem examinations (parental animals):
- SACRIFICE
- Male animals: All surviving animals after 43 days of dosing.
- Maternal animals: All surviving animals on day 4 of lactation or in females that failed to deliver, 25 days after evidence of mating detected.
GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.
HISTOPATHOLOGY / ORGAN WEIGHTS: The brain and ovaries were weighed in females. The ovaries, lungs, liver, kidneys, sciatic nerve, and gross lesions were examined histologically. The brain, testes, and epididymides were weighed in males. The testes, epididymides, accessory sex organs, liver, kidneys, sciatic nerve, and gross lesions were examined histologically. - Postmortem examinations (offspring):
- SACRIFICE: All F1 animals were sacrificed on lactation day 4.
- These animals were subjected to postmortem examinations macroscopic examination as follows:
GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.
- Statistics:
- Continuous data, including body weights, body weight gain, food consumption, organ weights, pup body weights, gestation length, mean live litter size and implantation scare counts were analyzed by One Way Analysis of Variance (ANOVA). If significance was detected, group by group comparisons were performed using Dunnett's Test. Count data were analyzed utilizing Chi-Square Test for copulation and fertility indices, pup sex ratios, the number of live and dead pups per group on lactation Day 0 and pup survival after lactation Day 0. All analyses utilized two-tailed tests for a minimum significance level of 5% comparing the control group to the treated groups.
- Reproductive indices:
- Fertility, gestation, parturition, and lactation
- Offspring viability indices:
- Live and dead pups, number of litters with live offspring, mean live litter size, male to female ratio, and the number of survivors
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No overt clinical signs of toxicity were noted in males or females from any of the study groups. Incidental findings were observed sporadically throughout the treated and control groups, and included dark material around the nose, scabbing, and hairloss (primarily on the forelimbs).
- Mortality:
- no mortality observed
- Description (incidence):
- All males survived to scheduled euthanasia on study Day 44. One female in the 500 mg/Kg/day group failed to deliver and was euthanized 25 days after evidence of mating was detected. The female was found to be gravid, and contained one autolyzed pup. All other females survived to study termination on lactation Day 4.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- There were no statistically significant or biologically meaningful differences noted in the mean body weights or body weight gains. Mean weights and weight gains for males and females were comparable amongst groups throughout the study.
- Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Male food consumtion, calculated as grams/animal/day and grams/Kg/day, was similar among the groups throughout the study; no statistical differences were noted.
Food consumption calculated as grams/Kg/day was statistically increased for females in the 100, 500, and 1000 mg/Kg/day groups during Gestation days 7-14, and for females in the 500 mg/Kg/day during Gestation days 14-20. These increases were not considered meaningful since similar statistical differences were not observed in grams/animal/day food consumption during these intervals. There were no statistical differences in female food consumption (grams/animal/day or grams/Kg/day) prior to mating or during lactation. - Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Test article-related microscopic changes were limited to the kidneys of males rats from the 100, 500, and 1000 mg/Kg/day groups. Most males rats in these groups had accumulations of hyaline droplets in the epithelial cells of the proximal convoluted tubules of the kidneys. There was a positive doseresponse relationship in the severity of this condition in the three treated groups. However, this condition is specific to young adult male rats and is of questionable toxicological significance.
Other microscopic lesions in the tissues and organs examined were considered spontaneous and not related to treatment with the test article. - Histopathological findings: neoplastic:
- not specified
- Other effects:
- not specified
- Reproductive function: oestrous cycle:
- not examined
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- Male and female copulation and fertility indices (i.e. pregnancy vs. succesful copulation), precoital intervals, amd gestation lengths were comparable among the groups; and the pregnancy rate was 100% in each group. There were no signs of prolonged delivery or unusual nesting behaviors noted in any of the study groups. However, one female in the 500 mg/Kg/day group was euthanized on post-coital day 25 and was found to contain one autolyzed pup.
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 1 000 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: Systemic and Reproductive Toxicity
- Key result
- Critical effects observed:
- no
- Clinical signs:
- no effects observed
- Mortality / viability:
- no mortality observed
- Description (incidence and severity):
- The number of live and dead pups, the number of litters with live offspring, the mean live litter size and the male to female pup ratio were comparable among the groups on lactation Day 0. Pup survival was comparable among the groups on lactation days 1 and 4.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- Mean pup weights were comparable among the groups on lactation days 0 and 4.
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- No remarkable findings were observed at necropsy in pups found dead prior to lactation day 4, or pups euthanized at study termination on lactation day 4. Incidental findings common to developing rat offspring were noted in each group, including controls.
- Other effects:
- no effects observed
- Description (incidence and severity):
- External Observations: There were no unusual external observations noted in the pups during lactation days 0 to 4. Incidental findings commonly observed in young offspring occurred in each group, including controls.
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 1 000 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: Developmental Toxicity
- Key result
- Critical effects observed:
- no
- Key result
- Reproductive effects observed:
- no
- Conclusions:
- The NOAEL for systemic, reproductive, and developmental toxicity was reported as 1000 mg/Kg/day, which excluded the hydrocarbon nephropathy noted in males.
- Executive summary:
In this screening for reproductive/developmental toxicity study, 1 -hexene was administered via gavage to twelve Sprague-Dawley rats/sex/dose at doses of 0, 100, 500, or 1000 mg/kg/day. The test material was composed of equal amounts of Neodene 6 alpha olefin, hexene-1 Gulftene 6, and alpha olefin C6 1 -hexene, which were obtained from different sources. Males were treated for 44 days beginning 28 days prior to mating and females were treated for 41 to 55 days beginning 14 days prior to mating through lactation day 4.
No reproductive or developmental effects were observed. There was a slight, but significant, decrease in absolute epididymal weight at all concentrations. The relative epididymal to brain weight was only significantly decreased in the low-dose group. Although the absolute epididymides weight was significantly decreased in parental males; the change was within 10% of the control, there was no dose response, there was no effects noted microscopically, and there were no effects on fertility. Therefore, this is not considered to be toxicologically significant. Pitted kidneys were observed at necropsy for 2 of 12 mid-dose males and 3 of 12 high-dose males. The predominant microscopic finding in males was the presence of large hyaline droplets in the proximal convoluted tubule that was dose related. These findings suggest hydrocarbon nephropathy, which is a toxicological effect specific to male rats and is not considered relevant to humans. There was no LOAEL for this study. The NOAEL for systemic, reproductive, and developmental toxicity was 1000 mg/kg/day, which excluded the hydrocarbon nephropathy in males.
This study received a Klimisch score of 1 and is classified as reliable without restrictions because it was conducted according to OECD 421 guidelines and was GLP compliant.
Referenceopen allclose all
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 ingroup 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.
No significant differences were detected for corpora lutea, implantation counts, implantation losses, litter size or litter viability for treated animals when compared to controls. Statistical analysis of the data did not reveal any significant intergroup differences.
OFFSPRING GROWTH and DEVELOPMENT
There were no treatment related effects detected. No obvious clinical signs of toxicity were detected for offspring from treated females when compared to controls. The incidental clinical signs detected throughout the control and treated groups, consisting of small size, no milk in stomach, a physical injury, found dead or missing, were considered to be low incidence findings observed in offspring in studies of this type and were considered unrelated to test item toxicity.
NECROPSY
No treatment-related macroscopic abnormalities were detected for interim death or terminal kill offspring. The incidental findings observed were those occasionally observed in reproductive studies of this type and were considered to be unrelated to toxicity of the test item.
Table 1. Summary of Reproductive Performance - Groups Values |
||||
|
Dose Group (mg/kg bw/day |
|||
0 (Control) |
100 |
300 |
1000 |
|
Males |
|
|||
Initial group size |
12 |
12 |
12 |
12 |
Paired |
12 |
12 |
12 |
12 |
Failed to induce pregnancy in female partner |
1 |
1 |
0 |
0 |
Induced pregnancy in female partner |
12 |
12 |
12 |
12 |
Surviving to terminal necropsy |
12 |
12 |
12 |
12 |
|
||||
Females |
|
|||
Initial group size |
12 |
12 |
12 |
12 |
Paired |
12 |
12 |
12 |
12 |
Total litter loss |
0 |
0 |
0 |
2 |
Non-pregnant |
1 |
1 |
0 |
0 |
Rearing young to Day 5 of age |
11 |
11 |
12 |
10 |
Table 2. 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 3. 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 4. 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 5. Group Mean Hematological Values - Females |
|||
Group |
|
Hb (g/dL) |
RBC (1012/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 6. 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 7. 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 8. 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 9. 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 10. 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
Table 11. Group Mean Implantation Losses and Survival Indices Values |
||||||
Group |
|
Pre-Implantation Loss (%) |
Post-Implantation Loss (%) |
Live Birth Index (%) |
Viability Index (%) |
|
Control (0 mg/Kg bw/day) |
Mean |
2.7 |
11.7 |
100.0 |
97.5 |
|
S.D. |
5.0 |
11.9 |
0.0 |
8.2 |
||
N |
11 |
11 |
11 |
11 |
||
|
||||||
100 mg/Kg bw/day |
Mean |
6.7 |
9.0 |
98.5 |
99.1 |
|
S.D. |
11.9 |
9.9 |
5.0 |
3.0 |
||
N |
11 |
11 |
11 |
11 |
||
|
||||||
300 mg/Kg bw/day |
Mean |
2.3 |
13.1 |
100.0 |
98.0 |
|
S.D. |
4.6 |
15.0 |
0.0 |
5.1 |
||
N |
12 |
12 |
12 |
12 |
||
|
||||||
1000 mg/Kg bw/day |
Mean |
5.5 |
6.5 |
99.3 |
95.4 |
|
S.D. |
17.2 |
7.7 |
2.1 |
7.1 |
||
N |
10 |
10 |
10 |
10 |
||
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 ingroup 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.
Male food consumtion, calculated as grams/animal/day and grams/Kg/day, was similar among the groups throughout the study; no statistical differences were noted. Food consumption calculated as grams/Kg/day was statistically increased for females in the 100, 500, and 1000 mg/Kg/day groups during Gestation days 7-14, and for females in the 500 mg/Kg/day during Gestation days 14-20. These increases were not considered meaningful since similar statistical differences were not observed in grams/animal/day food consumption during these intervals. There were no statistical differences in female food consumption (grams/animal/day or grams/Kg/day) prior to mating or during lactation.
Organ Weights:
Absolute epididymide weights for males in the 100, 500, 1000 mg/Kg/day groups were slightly, but statistically lower than the control group. Epididymide weights relative to brain weights were also slightly lower than the controls for males in each treated group, however, the difference was only statistically
significant at the 100 mg/Kg/day level.
Absolute and relative testes weights were comparable between the groups.
Absolute ovary weights and ovary weights relative to brain weight were comparable between the groups.
Necropsy:
Pitted kidneys were observed at necropsy in 2 of 12 males in the 500 mg/Kg/day group, and 3 of 12 males in the 1000 mg/Kg/day group. The pitted kidneys were possibly related to the hydrocarbon nephropathy observed micoscopically in the male rats. Other gross necropsy findings for males were generally unremarkable.
Gross necropsy findings for females that delivered were generally unremarkable, and the mean implantation scar counts for these females were comparable among the groups. For one 500 mg/Kg/day female euthanized post-breeding Day 25 (#3267), gross necropsy findings included dark red mucoid contents in the uterus, with one elongated and partially autolyzed pup located in the left uterine horn extending to the left cervix.
Histopatholoy:
Test article-related microscopic changes were limited to the kidneys of males rats from the 100, 500, and 1000 mg/Kg/day groups. Most males rats in these groups had accumulations of hyaline droplets in the epithelial cells of the proximal convoluted tubules of the kidneys. There was a positive dose-response relationship in the severity of this condition in the three treated groups. However, this condition is specific to young adult male rats and is of questionable toxicological significance.
Other microscopic lesions in the tissues and organs examined were considered spontaneous and not related to treatment with the test article.
CLINICAL SIGNS (OFFSPRING): There were no clinical signs of toxicity in the offspring.
BODY WEIGHT (OFFSPRING): There were no changes in body weight.
GROSS PATHOLOGY (OFFSPRING): There were no treatment-related changes in gross pathology.
The presence of large hyaline drops noted in the kidneys of paternal males suggests hydrocarbon nephropathy, which is a toxicological effect specific to male rats and is not considered a biologically relevant endpoint in humans. Therefore, this endpoint is excluded in determining the NOAEL. Although the absolute epididymides weight was significantly decreased in parental males; the change was within 10% of the control, there was no dose response, there was no effects noted microscopically, and there were no effects on fertility. Therefore, this is not considered to be toxicologically significant.
Table 2. Summary of F0 Gestation Food Consumption Data |
|||||
|
Group |
1 |
2 |
3 |
4 |
Level |
0 mg/Kg/day |
100 mg/Kg/day |
500 mg/Kg/day |
1000 mg/Kg/day |
|
Day 0-7 |
Mean |
76 |
81 |
77 |
80 |
S.D. |
8.1 |
8.2 |
7.7 |
7.8 |
|
N |
12 |
12 |
12 |
12 |
|
|
|||||
Day 7-14 |
Mean |
70 |
78* |
79* |
77* |
S.D. |
9.3 |
6.1 |
5.0 |
6.4 |
|
N |
12 |
12 |
12 |
12 |
|
|
|||||
Day 14-20 |
Mean |
65 |
67 |
72** |
68 |
S.D. |
5.9 |
5.8 |
5.3 |
4.4 |
|
N |
12 |
12 |
12 |
12 |
|
*p<0.05
**p<0.01
Table 3. Summary of F0 Male Gross Necropsy Observations |
||||
Group |
1 |
2 |
3 |
4 |
Level |
0 mg/Kg/day |
100 mg/Kg/day |
500 mg/Kg/day |
1000 mg/Kg/day |
No. of animals in Each dose group |
12 |
12 |
12 |
12 |
No. of animals Examined at Scheduled Euthanasia |
12 |
12 |
12 |
12 |
No Remarkable Findings |
9 |
10 |
7 |
5 |
Kidneys |
|
|||
- Pitted |
0 |
0 |
2 |
3 |
Table 4. Summary of F0 Female Gross Necropsy Observations |
||||
Group |
1 |
2 |
3 |
4 |
Level |
0 mg/Kg/day |
100 mg/Kg/day |
500 mg/Kg/day |
1000 mg/Kg/day |
No. of animals in Each dose group |
12 |
12 |
12 |
12 |
No. of animals Examined at Scheduled Euthanasia |
12 |
12 |
11 |
12 |
No Remarkable Findings |
9 |
10 |
7 |
5 |
Uterus |
|
|||
- Implantation Scar (s) Present Mean |
15.7 |
15.3 |
15.4 |
14.4 |
S.D. |
1.9 |
1.6 |
1.2 |
2.9 |
Table 5. Summary of F0 Male Organ Weight Data (grams) |
|||||
|
Group |
1 |
2 |
3 |
4 |
Level |
0 mg/Kg/day |
100 mg/Kg/day |
500 mg/Kg/day |
1000 mg/Kg/day |
|
Epididymides |
Mean |
1.36 |
1.22** |
1.26* |
1.24** |
S.D. |
0.110 |
0.094 |
0.083 |
0.078 |
|
N |
12 |
12 |
12 |
12 |
|
*p<0.05
**p<0.01
Table 6. Summary of F0 Male Organ Weight to Brain Weight Data (grams / 100 grams) |
|||||
|
Group |
1 |
2 |
3 |
4 |
Level |
0 mg/Kg/day |
100 mg/Kg/day |
500 mg/Kg/day |
1000 mg/Kg/day |
|
Epididymides |
Mean |
60.048 |
54.943* |
56.268 |
56.266 |
S.D. |
4.3247 |
4.553 |
3.9084 |
3.4050 |
|
N |
12 |
12 |
12 |
12 |
|
*p<0.05
Table 7. Histopathology Results |
||||||||
Tissue |
0 mg/Kg/day |
100 mg/Kg/day |
500 mg/Kg/day |
1000 mg/Kg/day |
0 mg/Kg/day |
100 mg/Kg/day |
500 mg/Kg/day |
1000 mg/Kg/day |
Male |
Female |
Male |
Female |
Male |
Female |
Male |
Female |
|
Kidney |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
12 |
Within normal limits |
4 |
6 |
2 |
7 |
3 |
7 |
0 |
6 |
Hyaline droplets, proximal convoluted tubules |
0 |
0 |
7 |
0 |
8 |
0 |
9 |
0 |
- minimal |
0 |
0 |
4 |
0 |
2 |
0 |
4 |
0 |
- mild |
0 |
0 |
1 |
0 |
6 |
0 |
1 |
0 |
- moderate |
0 |
0 |
2 |
0 |
0 |
0 |
3 |
0 |
- marked |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
Basophilic tubules, cortex |
2 |
2 |
0 |
0 |
0 |
0 |
3 |
0 |
- minimal |
2 |
2 |
0 |
0 |
0 |
0 |
3 |
0 |
Cyst |
0 |
0 |
0 |
1 |
0 |
0 |
2 |
1 |
- mild |
0 |
0 |
0 |
1 |
0 |
0 |
2 |
1 |
Hydronephrosis |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
- moderate |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
Mineralization |
0 |
4 |
0 |
3 |
0 |
4 |
0 |
4 |
- minimal |
0 |
3 |
0 |
2 |
0 |
4 |
0 |
4 |
- mild |
0 |
1 |
0 |
1 |
0 |
0 |
0 |
0 |
Nephritis, interstitial, chronic |
5 |
2 |
9 |
2 |
4 |
1 |
9 |
3 |
- minimal |
5 |
1 |
9 |
2 |
3 |
1 |
5 |
3 |
- mild |
0 |
1 |
0 |
0 |
1 |
0 |
4 |
0 |
Pyelitis, chronic |
1 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
- moderate |
1 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
Pyelonephritis, acute |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
- moderate |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
Effect on fertility: via oral route
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- NOAEL
- 300 mg/kg bw/day
- Study duration:
- subchronic
- Species:
- rat
- Quality of whole database:
- In total, nine good quality screening studies are available on nine category members, covering C6 to C18 higher olefins. The study on nonene, branched, in common with all other studies, showed no effects on fertility at levels up tp 1000 mg/Kg. There was however, a slight effect on post-natal pup viability, litter size and weight at 1000 mg/Kg, with a clear NOAEL of 300 mg/Kg. This study was selected as being the 'worst case', although it should be recognized that none of the other eight studies showed any reproductive effects on post-natal pup viability.
Effect on fertility: via inhalation route
- Endpoint conclusion:
- no study available
Effect on fertility: via dermal route
- Endpoint conclusion:
- no study available
Additional information
A guideline Oral (Gavage) Combined Repeat Dose Toxicity Study with Reproduction/Developmental Toxicity Screening Test in the Rat (OECD 422) was performed on Oct-1-ene (CAS # 111 -66 -0; Harlan Laboratories, 2014).
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 differences between treated and control animal were detected in body weight development and food consumption. In contrast, 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, and the ‘No Observed Effect Level’ (NOEL) for reproductive toxicity was considered to be 1000 mg/Kg bw/day.
In a supporting Guideline (OECD 422) combined repeated dose, reproductive/developmental toxicity screening test (Harlan Laboratories, 2014), the test material (Decene; CAS# 25339-53-1) was administered by gavage to three groups, each of twelve male and twelve female Wistar Han™:RccHan™:WIST strain rats, for up to eight weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females), at dose levels of 100, 300 and 1000 mg/Kg bw/day. A control group of twelve males and twelve females was dosed with vehicle alone (Arachis oil BP).
Clinical signs, behavioural assessments, body weight change and food and water consumption were monitored during the study.
Pairing of animals within each dose group was undertaken on a one male: one female basis within each treatment group on Day 15 of the study, with females subsequently being allowed to litter and rear their offspring to Day 5 of lactation.
During the lactation phase, daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of surface righting reflex. Extensive functional observations were performed on five selected males from each dose group after the completion of the pairing phase, and for five selected parental females from each dose group on Day 4 post-partum. Haematology and blood chemistry were evaluated prior to termination on five selected males and females from each dose group.
Surviving adult males were terminated on Day 43, followed by the termination of all females and surviving offspring on Day 5 post-partum. Any female which did not produce a pregnancy was terminated on or after Day 25 post coitum. Two females showing no visible vaginal opening were terminated on Day 42. All animals were subjected to a gross necropsy examination and histopathological evaluation of selected tissues was performed.
No test material related mortality was observed through the study period. Oral gavage administration of the test material at dosages of 100, 300 and 1000 mg/Kg bw/day for approximately six weeks resulted in minimal to moderate microscopic changes in the fore stomach of one male at 300 mg/Kg bw/day and both sexes at 1000 mg/Kg bw/day. These minimal to moderate histopathological changes in the fore stomach were considered to have occurred due to a local irritant effect, rather than true systemic toxicity. As a consequence, the NOAEL for adult systemic toxicity was considered to be 1000 mg/Kg bw/day.
There were no treatment-related effects on mating performance; fertility; or gestation length apparent at 100, 300 or 1000 mg/Kg bw/day. There was no effect of treatment on corpora lutea count, pre-implantation loss, numbers of implantations, post-implantation loss, litter size, sex ratio and subsequent offspring survival to Day 4 of age at 100, 300 or 1000 mg/Kg bw/day. Offspring body weight or body weight gain and litter weights, surface righting ability on Day 1 or clinical signs to Day 5 of age remained unaffected post exposure at 100, 300 or 1000 mg/Kg bw/day. Additionally, gross necropsy findings for offspring did not indicate any adverse effect of maternal treatment on offspring development at 100, 300 or 1000 mg/Kg bw/day.
Based on the results observed, the No Observed Effect Level (NOEL) for reproduction, including the survival, growth and development of the offspring, is considered to be 1000 mg/Kg bw/day.
Effects on developmental toxicity
Description of key information
Guideline (OECD 414) developmental toxicity studies have been conducted for five members of the higher olefins category.
No effects of the development of the off-spring were observed in any study at dose levels up to 1000 mg/Kg bw/day.
Read-across of these results to Oct-2 -ene is claimed as valid based on the justifications provided for both analogue and category approaches
Link to relevant study records
- Endpoint:
- developmental toxicity
- 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:
- other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Oct-1-ene
- Cs No.: 111-66-0
- Purity test date: >98%
- Lot/batch No.: 719238
- Expiration date of the lot/batch: 29 April 2014
- Label: Alpha Olefin C8 Lot 719238
- Data received: 29 April 2013
- Storage condition of test material: room temperature in the dark under nitrogen
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
- 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 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.
- Details on mating procedure:
- - M/F ratio per cage: 1/1 (pairing period)
- Length of cohabitation: minimum 14 days
- Proof of pregnancy: The presence of sperm within the vaginal smear and/or vaginal plug in situ was taken as positive evidence of mating (Day 0 of gestation)
- After successful mating each pregnant female was caged: males were returned to their original holding cages (unless required for additional pairing). Mated females were housed individually during the period of gestation and lactation. - 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. - Maternal examinations:
- 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 AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Water intake was measured daily during the pre-pairing phase of the
BEHAVIORAL, FUNCTIONAL OBSERVATIONS and SENSORY REACTIVITIES were observed and recorded.
Postmortem Parental:
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. - Fetal examinations:
- Number of offspring born
Number of offspring alive recorded daily and reported on Days 1 and 4 post partum
Sex of offspring on Days 1 and 4 post partum
Clinical condition of offspring from birth to Day 5 post partum
Individual offspring weights on Days 1 and 4 post partum (litter weights were calculated retrospectively from this data)
Postmortem Offspring:
All adult animals and offspring, including those dying during the study, were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded. - 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 differences were 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
- Other effects:
- no effects observed
- Description (incidence and severity):
- Reproductive performance
Mating: There were no treatment-related effects on mating performance.
Fertility: No treatment-related effects on fertility were detected for treated animals, when compared to controls. One control female and one female treated with 100 mg/kg bw/day did not achieve pregnancy following evidence of mating. No histopathological correlates were evident in the reproductive organs for these animals and in the absence of any similar infertility effects detected at 1000 mg/kg bw/day the intergroup differences were considered not to be related to test item toxicity.
Gestation Length: There were no differences in gestation lengths. The distribution for treated females was comparable to controls. The gestation lengths were between 22 and 23½ days. - Details on maternal toxic effects:
- 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 outs ide the background range, in the absence of any associated histology correlates all the intergroup differences were 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.
- 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.
- Basis for effect level:
- other: Systemic toxicity
- Key result
- Dose descriptor:
- NOEL
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: Reproductive Toxicity
- Key result
- Abnormalities:
- no effects observed
- Fetal body weight changes:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Reduction in number of live offspring:
- no effects observed
- External malformations:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Other effects:
- no effects observed
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:no effects
Details on embryotoxic / teratogenic effects:
OFFSPRING LITTER SIZE, SEX RATIO and VIABILITY
No significant differences were detected for corpora lutea, implantation counts, implantation losses, litter size or litter viability for treated animals when compared to controls. Statistical analysis of the data did not reveal any significant intergroup differences.
OFFSPRING GROWTH and DEVELOPMENT
There were no treatment related effects detected. No obvious clinical signs of toxicity were detected for offspring from treated females when compared to controls. The incidental clinical signs detected throughout the control and treated groups, consisting of small size, no milk in stomach, a physical injury, found dead or missing, were considered to be low incidence findings observed in offspring in studies of this type and were considered unrelated to test item toxicity.
NECROPSY
No treatment-related macroscopic abnormalities were detected for interim death or terminal kill offspring. The incidental findings observed were those occasionally observed in reproductive studies of this type and were considered to be unrelated to toxicity of the test item. - Key result
- Dose descriptor:
- NOEL
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: Developmental Toxicity
- Key result
- Abnormalities:
- no effects observed
- Key result
- Developmental effects observed:
- no
- Conclusions:
- The ‘No Observed Effect Level’ (NOEL) for reproductive/developmental toxicity was considered to be 1000 mg/Kg bw/day.
- 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.
No significant differences were detected for corpora lutea, implantation counts, implantation losses, litter size or litter viability for treated animals when compared to controls. Statistical analysis of the data did not reveal any significant intergroup differences. No obvious clinical signs of toxicity were detected for offspring from treated females when compared to controls. The incidental clinical signs detected throughout the control and treated groups, consisting of small size, no milk in stomach, a physical injury, found dead or missing, were considered to be low incidence findings observed in offspring in studies of this type and were considered unrelated to test item toxicity.
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 The ‘No Observed Effect Level’ (NOEL) for reproductive/developmental toxicity was considered to be 1000 mg/Kg bw/day.
- Endpoint:
- developmental toxicity
- 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
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Species:
- rat
- Analytical verification of doses or concentrations:
- yes
- Frequency of treatment:
- Daily
- Clinical signs:
- effects observed, treatment-related
- 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 differences were 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
- Other effects:
- no effects observed
- Description (incidence and severity):
- Reproductive performance
Mating: There were no treatment-related effects on mating performance.
Fertility: No treatment-related effects on fertility were detected for treated animals, when compared to controls. One control female and one female treated with 100 mg/kg bw/day did not achieve pregnancy following evidence of mating. No histopathological correlates were evident in the reproductive organs for these animals and in the absence of any similar infertility effects detected at 1000 mg/kg bw/day the intergroup differences were considered not to be related to test item toxicity.
Gestation Length: There were no differences in gestation lengths. The distribution for treated females was comparable to controls. The gestation lengths were between 22 and 23½ days. - Details on maternal toxic effects:
- 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 outs ide the background range, in the absence of any associated histology correlates all the intergroup differences were 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.
- 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.
- Basis for effect level:
- other: Systemic toxicity
- Key result
- Dose descriptor:
- NOEL
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: Reproductive Toxicity
- Key result
- Abnormalities:
- no effects observed
- Fetal body weight changes:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Reduction in number of live offspring:
- no effects observed
- External malformations:
- no effects observed
- Description (incidence and severity):
- There were no treatment related effects detected.
- Other effects:
- no effects observed
- Details on embryotoxic / teratogenic effects:
- Embryotoxic / teratogenic effects:no effects
Details on embryotoxic / teratogenic effects:
OFFSPRING LITTER SIZE, SEX RATIO and VIABILITY
No significant differences were detected for corpora lutea, implantation counts, implantation losses, litter size or litter viability for treated animals when compared to controls. Statistical analysis of the data did not reveal any significant intergroup differences.
OFFSPRING GROWTH and DEVELOPMENT
There were no treatment related effects detected. No obvious clinical signs of toxicity were detected for offspring from treated females when compared to controls. The incidental clinical signs detected throughout the control and treated groups, consisting of small size, no milk in stomach, a physical injury, found dead or missing, were considered to be low incidence findings observed in offspring in studies of this type and were considered unrelated to test item toxicity.
NECROPSY
No treatment-related macroscopic abnormalities were detected for interim death or terminal kill offspring. The incidental findings observed were those occasionally observed in reproductive studies of this type and were considered to be unrelated to toxicity of the test item. - Key result
- Dose descriptor:
- NOEL
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: Developmental Toxicity
- Key result
- Abnormalities:
- no effects observed
- Key result
- Developmental effects observed:
- no
- Conclusions:
- An Oral (Gavage) Combined Repeat Dose Toxicity Study with Reproduction/Developmental Toxicity Screening Test in the Rat (OECD 422) was performed on the read-across source material Oct-1-ene CAS 111-66-0. The ‘No Observed Effect Level’ (NOEL) for reproductive/developmental toxicity was considered to be 1000 mg/Kg bw/day.
Read-across of this result to Oct-2 -ene is claimed as valid based on the justifications provided for both analogue and category approaches - 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 read-across source 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.
No significant differences were detected for corpora lutea, implantation counts, implantation losses, litter size or litter viability for treated animals when compared to controls. Statistical analysis of the data did not reveal any significant intergroup differences. No obvious clinical signs of toxicity were detected for offspring from treated females when compared to controls. The incidental clinical signs detected throughout the control and treated groups, consisting of small size, no milk in stomach, a physical injury, found dead or missing, were considered to be low incidence findings observed in offspring in studies of this type and were considered unrelated to test item toxicity.
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 The ‘No Observed Effect Level’ (NOEL) for reproductive/developmental toxicity was considered to be 1000 mg/Kg bw/day.
Read-across of this result to Oct-2 -ene is claimed as valid based on the justifications provided for both analogue and category approaches.
Referenceopen allclose all
Table 1. Summary of Reproductive Performance - Groups Values |
||||
|
Dose Group (mg/kg bw/day |
|||
0 (Control) |
100 |
300 |
1000 |
|
Males |
|
|||
Initial group size |
12 |
12 |
12 |
12 |
Paired |
12 |
12 |
12 |
12 |
Failed to induce pregnancy in female partner |
1 |
1 |
0 |
0 |
Induced pregnancy in female partner |
12 |
12 |
12 |
12 |
Surviving to terminal necropsy |
12 |
12 |
12 |
12 |
|
||||
Females |
|
|||
Initial group size |
12 |
12 |
12 |
12 |
Paired |
12 |
12 |
12 |
12 |
Total litter loss |
0 |
0 |
0 |
2 |
Non-pregnant |
1 |
1 |
0 |
0 |
Rearing young to Day 5 of age |
11 |
11 |
12 |
10 |
Table 2. 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 3. 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 4. 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 5. Group Mean Hematological Values - Females |
|||
Group |
|
Hb (g/dL) |
RBC (1012/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 6. 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 7. 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 8. 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 9. 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 10. 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
Table 11. Group Mean Implantation Losses and Survival Indices Values |
||||||
Group |
|
Pre-Implantation Loss (%) |
Post-Implantation Loss (%) |
Live Birth Index (%) |
Viability Index (%) |
|
Control (0 mg/Kg bw/day) |
Mean |
2.7 |
11.7 |
100.0 |
97.5 |
|
S.D. |
5.0 |
11.9 |
0.0 |
8.2 |
||
N |
11 |
11 |
11 |
11 |
||
|
||||||
100 mg/Kg bw/day |
Mean |
6.7 |
9.0 |
98.5 |
99.1 |
|
S.D. |
11.9 |
9.9 |
5.0 |
3.0 |
||
N |
11 |
11 |
11 |
11 |
||
|
||||||
300 mg/Kg bw/day |
Mean |
2.3 |
13.1 |
100.0 |
98.0 |
|
S.D. |
4.6 |
15.0 |
0.0 |
5.1 |
||
N |
12 |
12 |
12 |
12 |
||
|
||||||
1000 mg/Kg bw/day |
Mean |
5.5 |
6.5 |
99.3 |
95.4 |
|
S.D. |
17.2 |
7.7 |
2.1 |
7.1 |
||
N |
10 |
10 |
10 |
10 |
||
Effect on developmental toxicity: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 1 000 mg/kg bw/day
- Study duration:
- subchronic
- Species:
- rat
- Quality of whole database:
- In total, fourteen good quality guideline and screening studies are available for ten category members, covering C6 to C30 higher olefins. The study on Octadecene is one of five good quality, guideline reproductive toxicity studies conducted on members of the higher olefins category. In addition, nine supporting toxicity screening studies are available for category members. None of the studies showed any evidence of effects on development of the rat foetus at levels up to 1000 mg/Kg.
Effect on developmental toxicity: via inhalation route
- Endpoint conclusion:
- no study available
Effect on developmental toxicity: via dermal route
- Endpoint conclusion:
- no study available
Additional information
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 (Harlan Laboratories Ltd., 2014).
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.
No significant differences were detected for corpora lutea, implantation counts, implantation losses, litter size or litter viability for treated animals when compared to controls. Statistical analysis of the data did not reveal any significant intergroup differences. No obvious clinical signs of toxicity were detected for offspring from treated females when compared to controls. The incidental clinical signs detected throughout the control and treated groups, consisting of small size, no milk in stomach, a physical injury, found dead or missing, were considered to be low incidence findings observed in offspring in studies of this type and were considered unrelated to test item toxicity.
In view of these results, the NOAEL ‘for systemic toxicity was considered to be 1000 mg/Kg bw/day The ‘No Observed Effect Level’ (NOEL) for reproductive/developmental toxicity was considered to be 1000 mg/Kg bw/day.
Justification for classification or non-classification
Guideline screening reproductive toxicity (OECD 421 and 422) and developmental toxicity (OECD 414) studies have been conducted for members of the Higher Olefins category covering C6 to C18.
The weight of evidence from oral reproductive and developmental toxicity studies, accompanied with data from oral and inhalation sub-chronic toxicity studies in rats indicate that category members have little or no potential to be considered reproductive/developmental toxicants.
Therefore, no classification is considered necessary under the current CLP regulation.
Read-across of this result to Oct-2 -ene is claimed as valid based on the justifications provided for both analogue and category approaches.
Additional information
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
