Oct-2-ene

Developmental toxicity / teratogenicity

Administrative data

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.

Cross-reference

Reason / purpose for cross-reference:

read-across: supporting information

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 001

Cas Number:

111-66-0

Molecular formula:

C8H16

Test material form:

other: clear colorless liquid

Details on test material:

- 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

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.

Test animals

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

Administration / exposure

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

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

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

Remarks:

Control

Doses / concentrations 2

Dose / conc.:

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

Doses / concentrations 3

Dose / conc.:

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

Doses / concentrations 4

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.

Examinations

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).

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

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

Maternal developmental toxicity

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.

Effect levels (maternal animals)

1

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

2

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

3

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

Maternal abnormalities

Key result

Abnormalities:

no effects observed

Results (fetuses)

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.

Effect levels (fetuses)

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

Fetal abnormalities

Key result

Abnormalities:

no effects observed

Overall developmental toxicity

Key result

Developmental effects observed:

no

Any other information on results incl. tables

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

* Significantly different from control group p<0.05

Table 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

Applicant's summary and conclusion

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.