Registration Dossier

Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11-27-2013 to 03-09-2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: test substance was well characterized and the study was conducted according to guidelines without significant deviation

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
Additional histopathological assessment of the reproductive organs
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Density 0.8581 g/cm3
Storage Conditions: At ambient temperature, protected from light

Test animals

Species:
rat
Strain:
other: Han Wistar (CRL:WI(Han))
Sex:
male/female
Details on test animals and environmental conditions:
At the initiation of dosing the animals were approximately 7-8 weeks old and weighed between 193-254 g (males) and 139-196 g (females).

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5% HPMC (Methocel E4M), 0.1% Tween 80 in Milli-Q water
Details on oral exposure:
The test and control items were administered to the appropriate animals by once daily oral
gavage for at least 90 days, using a syringe with attached gavage cannula.
The dose volume (at 10 mL/kg bw) for each animal was based on the most recent body weight measurement.
The first day of dosing for each animal was designated as Day 1. The dosing formulations were stirred for at least 30 minutes before and continuousyduring dose administration. On completion of the dosing period the designated recovery animals were retained for
28 days for assessment of recovery.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analyses were performed using a validated analytical procedure (A.P. No.3113.02, validated under Charles River Study No. 431130). Analytical variation of dose formulations was within the accepted range.

Concentration and Homogeneity Analysis
Duplicate 1 mL top, middle and bottom samples (middle only for the control group) were
taken into Falcon tubes, see Appendix 1, for each sampling time point and sent to the
analytical laboratory; triplicate top, middle and bottom samples (1 mL in Falcon tubes) were
retained at the Test Facility as backup samples. Concentration results were considered
acceptable if sample concentration results within or equal to ± 15% of theoretical
concentration and each individual sample concentration result within or equal to ± 20%.
For homogeneity, the criteria for acceptability were a relative standard deviation (RSD) of
concentrations of ≤ 10% for each group.

Stability Analysis
Stability analyses performed previously in conjunction with Charles River Study No. 431130
demonstrated that the test item is stable in the vehicle when prepared and stored under the
same conditions at concentrations bracketing those used in the present study. Stability data
have been retained in the study records for Charles River Study No. 431130.





Duration of treatment / exposure:
90 days
Frequency of treatment:
once daily
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
100 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
300 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10

5 males and 5 females for each of the recovery groups
Control animals:
yes, concurrent vehicle
Details on study design:

The dose levels were agreed with the Sponsor after evaluation of a preliminary study carried out by Charles River (Charles River Study No. 525506).

- Post-exposure recovery period in satellite groups: two groups (control and 1000 mg/kg bw/day) for a 28d recovery period
Positive control:
none

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
Mortality / Moribundity check: twice daily (in the morning and as late as practical each day)
Additional clinical observations: examination for reaction to treatment particular during and for the first hour after dosing.

Detailed Clinical Observations: Yes
All animals were removed from their cages and subjected to a detailed clinical examination
once a week from Week -1 and on the first day of scheduled necropsy.

Postdose Observations
All animals were observed regularly throughout the day during the dosing period for reaction
to treatment. Particular attention was paid to the animals during and for the first hour after
dosing and the onset, intensity and duration of any observations were recorded.
In addition, all animals were observed at least once daily during the recovery period.

Body Weights: yes
Body weights were recorded for each animal twice during the pretreatment period, daily
during the dosing period and twice weekly during the recovery period and on the first day of
scheduled necropsy.
Body weights recorded immediately before dosing were classified on the in-life data capture
system as Day 0 body weights, relating to the number of days of treatment completed.
Subsequent body weights also followed this pattern.

Food Consumption: Yes
Food consumption was quantitatively measured once during the pretreatment period, weekly
during the dosing period (over 7 days in Weeks 1 to 12 and over 6 days in Week 13) and
recovery period.
Food consumption recorded immediately before dosing were classified on the in-life data
capture system as Day 0 and represents food consumption measured from previous data
collection time point (Day -7) to Day 0. Subsequent food consumption also followed this
pattern.

Ophthalmic Examinations: yes
All animals were subject to an ophthalmic examination during the pretreatment period,
Control and High dose animals were examined in Week13 of the dosing period, and all
recovery animals were examined during Week 17.
The anterior, lenticular and fundic areas of the eyes were examined using an indirect
ophthalmoscope, after the application of a mydriatic agent (1% Tropicamide, Mydriacyl®).

Detailed Functional Observations: Yes
Detailed functional observations were performed for all animals once during the pretreatment
period and during Week 12 of the dosing period and on all recovery animals during Week 17.
Examinations were performed at an approximately standard time of day and were conducted
by a technician who was not involved in the dosing procedures or in the collection of body
weight and food consumption data, see Appendix 1. The cage cards showing treatment group
were removed from each cage before the independent technician entered the room to perform
the examinations, the second pre-prepared card were left as the functional observation animal
identifier. In each cage one or two animals had their tails marked in order to allow the
independent technician to identify each animal.

Cageside Observations
On first approach to the cage, the technician checked the posture/condition of the animal for
signs of prostration, lethargy, writhing, circling, breathing abnormalities, gait abnormalities,
tremors, fasciculation, convulsions, biting (of cage components or self-mutilation),
vocalisations and piloerection.
Body temperature was recorded from the electronically implanted microchip. Where the
microchip failed to record an animal’s temperature onto the data capture system, a rectal
temperature was recorded.
The technician also checked the ease at which the animal was removed from the cage, the
condition of its eyes (for pupillary function, miosis, mydriasis, exophthalmos, encrustation
and lacrimation), the condition of the coat, the presence of salivation and the overall ease of
handling.

Observations in a standardised arena (2 min observation period):
Animals were checked for latency (time to first locomotory movement), level of mobility,
rearing, grooming, urination/defecation, arousal (level of alertness), posture,
tremor/convulsions, vocalisation, piloerection, palpebral closure, gait abnormalities and
stereotypy and/or unusual behaviours.

Functional Tests: yes
The technician observed the reactions to a sudden sound (click above the head) and the
reaction to touch on the rump with a blunt probe.

Grip strength:
This was measured using a Dual/Single Channel Meter (Linton Instruments), to which was
attached a wire screen assembly. Once the animal had gripped the screen, the body was
pulled until its grasp was broken; the strain gauge recorded the force required. The procedure
was repeated 3 times for the forelimbs and 3 times for the hindlimbs, and the mean fore and
hind grip strengths calculated.

Pain perception:
This was assessed by measurement of the tail flick response, using a technique based on the
method devised by D`Amour and Smith (1941). The apparatus used shone a calibrated infrared
heat source onto the tail and automatically measured the reaction time of the animal
(accurate to 0.1 s). It was ensured that no visible injury to the tail was caused during this test.

Landing Foot Splay:
Maize oil was applied to the hind paws of each animal. The animal was then held in a
horizontal, prone position with the nose ca 30 cm above a bench surface covered with
absorbent paper. When the animal was calm, it was dropped. The distance between the
prints of the central footpads was measured and the average measurement recorded. The
procedure was repeated 3 times. If the rat did not land properly on its feet, this was recorded.

Motor activity:
Each animal was placed in an individual cage held within a Smartfame utilising infra-red
pyroelectric detectors. Movement was detected in 2 dimensions anywhere in the cage, and
was differentiated into basic and fine movements, and X and Y ambulation. Each animal was
monitored for one session of 1 h, activity counts being recorded over successive period of 5
min each.
Any other abnormality not already recorded in the above screening battery.

Haematology: yes
Blood samples (0.5 mL) were taken in to EDTA and analysed for the parameters specified below:

Haematology Parameters
Red blood cell count
Haemoglobin
Haematocrit
Mean cell volume
Mean cell haemoglobin concentration
Mean cell haemoglobin
Reticulocytes
Reticulocyte count (absolute)
Red blood cell distribution width
Platelets
White blood cell count
Neutrophils
Lymphocytes
Monocytes
Eosinophils
Basophils
Large unstained cells
Other cells (as appropriate)

A blood smear was prepared from each haematology sample, labelled, stained, and stored
without evaluation.

Coagulation
Blood samples (0.9 mL) were taken in to 3.8% (w/v) trisodium citrate and processed for
plasma, which was analysed for the parameters listed below
Coagulation
Parameters
Activated partial thromboplastin time
Fibrinogen
Prothrombin time

Clinical Chemistry
Blood samples (1.0 mL) were taken in to lithium heparin and processed for plasma, which
was analysed for the parameters specified below:

Clinical Chemistry Parameters
Urea
Glucose
Aspartate aminotransferase
Alanine aminotransferase
Alkaline phosphatase
Creatine phosphokinase
Lactate dehydrogenase
Sodium
Potassium
Chloride
Total protein
Albumin
Globulin
Albumin/globulin ratio
Cholesterol
Creatinine
Total bilirubin
Calcium
Inorganic phosphate

Urinalysis
Urine samples were processed and analysed for the parameters listed in Text Table 8.
Text Table below:
Urinalysis Parameters
Microscopic evaluation of spun deposit
Colour
Turbidity
Specific gravity
Volume
pH
Protein
Glucose
Bilirubin
Ketones
Leukocytes
Blood Pigments
Urobilinogen
Sacrifice and pathology:
All main study and recovery animals were subjected to a complete necropsy examination,
which included evaluation of the carcass and musculoskeletal system; all external surfaces
and orifices; cranial cavity and external surfaces of the brain; and thoracic, abdominal, and
pelvic cavities with their associated organs and tissues. A veterinary pathologist was
available for consultation during scheduled necropsy examinations.

The organs (see list below) were weighed at necropsy for all scheduled euthanasia
animals. Organ weights were not recorded for animals euthanised in poor condition or in
extremis. Paired organs were weighed separately but are reported together. Terminal body
weights were used for organ weight analysis.

Organs weighted at necropsy:
Epididymis
Gland, adrenal
Gland, pituitary
Gland, prostate
Gland, thyroid
Heart
Kidney
Liver
Lung
Ovary
Spleen
Testis
Thymus
Uterus

HISTOPATHOLOGY: Yes. Tissues identified in Table 1, with the exception of testes, were processed at the Test Site. Testes samples were fixed in Modified Davidson’s solution for up to 72 hours and then trimmed and processed to wax impregnation at the Test Facility. Further processing of testes and entire processing of all other tissue samples required was carried out at the Test Site. All other tissues were embedded in paraffin, sectioned, mounted on glass slides, and stained with haematoxylin and eosin.
Histopathological evaluation was performed by a board-certified veterinary pathologist with training and experience in laboratory animal pathology.


Other examinations:
Two bone marrow smears were prepared from each euthanised animal, air dried, fixed in methanol, stained
with May-Grunwald-Giemsa stain, and cover slipped. Bone marrow smears were not evaluated.
Statistics:
Unless otherwise stated, all statistical tests were two-sided and performed at the 5%
significance level using in-house software. Males and females were analysed separately.
The following pairwise comparisons were performed:
Control Group vs Group 2
Control Group vs Group 3
Control Group vs Group 4
Body weight, food consumption (only for main study animals), selected functional
observational battery and motor activity data, haematology, coagulation, clinical chemistry
and selected urinalysis data were analysed for homogeneity of variance using the ‘F-Max'
test. Where the group variances appeared homogeneous, a parametric ANOVA was used and
pairwise comparisons were made using Fisher’s F protected LSD method via Student's t test
i.e. pairwise comparisons were made only if the overall F-test was significant. Where the
variances were heterogeneous, log or square root transformations were used in an attempt to
stabilise the variances. Where the variances remained heterogeneous, then a Kruskal-Wallis
non-parametric ANOVA was used and pairwise comparisons were made using chi squared
protection (via z tests, the non-parametric equivalent of Student's t test).
In circumstances where it was not possible to perform the F Max test due to zero standard
deviation in at least one group, the non-parametric ANOVA results are reported.
Organ weights were analysed using ANOVA as above and by analysis of covariance
(ANCOVA) using terminal kill body weight as covariate. In addition, organ weights as a
percentage of terminal body weight were analysed using ANOVA as above as an exploratory
analysis.
In circumstances where the variances in the ANCOVA remained heterogeneous following
log or square root transformations, the data were subjected to a rank transformation prior to
analysis.

In the ANOVA and ANCOVA summary tables, the results of the analysis are reported
indicating the level of statistical significance (p<0.05, p<0.01 and p<0.001) of each pairwise
comparison.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
reversible light increase liver weight values in females given 1000 mg/kg/day which did not show a dose-related trend. These changes were considered incidental; as there were no patterns, trends or or correlating data to suggest toxicological relevance.
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
There was one unscheduled death during the study that was not related to administration of the test item.
Animal 155F (0 mg/kg/day) was euthanised at Day 64 due to the presence of a ruptured subcutaneous mass on the ventral thorax. Microscopically, the ulcerated mass presented as an extensive area of chronic active inflammation in the subcutis, which was considered secondary to a gavage dosing accident. Increased extramedullary haemopoiesis in the spleen and an increased number of myeloid cells in the bone marrow of the sternum and the femur were secondary to the increased requirement for inflammatory cells. Microgranulomata were also seen in the liver of this animal.
There were no clinical signs that were considered to be attributable to administration of the test item, Alkenes, C11/C12, hydroformylation products, distillation residues (CAS/NR. 90622-27-8).
Animal 472F (Group 4, 1000 mg/kg/day) was noted with damaged/missing teeth from Day 68 to 74. Body weight loss, piloerection and hunched body posture were observed in this period. To avoid further body weight loss soft diet pallets were given to this animal.
These findings were considered unrelated to administration of Alkenes, C11/C12, hydroformylation products, distillation residues (CAS/NR. 90622-27-8). An excess salivation was recorded up to 15 minutes following dosing, in four males and two females receiving Alkenes, C11/C12, hydroformylation products, distillation residues (CAS/NR. 90622-27-8) at 300 mg/kg/day and all animals at 1000 mg/kg/day. There were incidences of ploughing recorded in two males and two females at 300 mg/kg/day or two males and four females at 1000 mg/kg/day. Incidences of stained fur were also recorded in a few animals from all treated groups. All these clinical observations were considered not to be adverse. An excess salivation and ploughing may have been observed due to the taste of the test item.
The other clinical observations recorded were considered incidental, of the nature commonly observed in this strain and age of rats, therefore, were considered unrelated to administration of Alkenes, C11/C12, hydroformylation products, distillation residues (CAS/NR. 90622-27-8).

BODY WEIGHT AND WEIGHT GAIN
No test item related changes in body weight occurred during the study.

OPHTHALMOSCOPIC EXAMINATION
All findings and incidences were considered to be representative of the normal background findings observed at the testing laboratory.

HAEMATOLOGY
There were no differences in haematology and coagulation parameters that were considered to be related to treatment.
Monocyte count was statistically significantly higher than control values in Week 13 in males at 300 and 1000 mg/kg/day and females at 1000 mg/kg/day. However, the differences were small, there was no dose relationship observed in males and the differences were no longer evident following recovery period. Consequently, these differences are not considered to be toxicologically significant. Activated partial thromboplastin time was statistically significantly shorter than control values in Week 13 in females at 100 and 300 mg/kg/day. Due to lack of changes in the related parameters and lack of clear dose relationship and no changes observed in animals at 1000 mg/kg/day, these differences are not considered to be toxicologically significant. Haematocrit, mean cell haemoglobin and mean cell volume were lower in females treated with Alkenes, C11/C12, hydroformylation products, distillation residues (CAS/NR. 90622-27-8) at 1000 mg/kg/day in Week 17, following 4 weeks recovery period. As these differences were not observed at the end of the treatment period and due to the small magnitude of changes these differences are not considered to be toxicologically significant.

CLINICAL CHEMISTRY
There were no differences in clinical chemistry parameters that were considered to be related to treatment.
Alkaline phosphatase was noted to be slightly higher in males at 100 and 1000 mg/kg/day in Week 13 and slightly lower in males at 1000 mg/kg/day in Week 17. Due to the small magnitude of change and lack of a dose relationship in Week 13 these differences are not considered to be toxicologically significant.
Creatinine level was noted to be slightly higher in males at 300 mg/kg/day in Week 13 when compared to the Controls. Due to the small magnitude of change and lack of a dose relationship this difference is not considered to be toxicologically significant.
Glucose level was noted to be slightly higher in females at 1000 mg/kg/day in Week 17 when compared to the Controls. As these differences were not observed on the end of the treatment period and due to the small magnitude of change and lack of a dose relationship this difference is not considered to be toxicologically significant.
Sodium level was noted to be slightly lower in females at 1000 mg/kg/day in Week 17 when compared to the Controls. As these differences were not observed on the end of the treatment period due to the small magnitude of change and lack of a dose relationship this difference is not considered to be toxicologically significant.

URINALYSIS
There were no differences in urinalysis parameters in either Week 13 or 17.

NEUROBEHAVIOUR
There were no differences between treated groups and controls in clinical signs seen during the functional observation battery recordings, or in the values recorded for foot splay, grip strength, tail flick and body temperature at either the pretrial or the Weeks 12 and 17 observations.
Although there were apparent separated occasions when differences were noted between motor activity of control and treated animals in Week 12 and Week 17, in the absence of changes in any other functional observation battery parameters and endpoints and clear dose relationship, such differences are not considered to be of toxicological significance and related to treatment.

ORGAN WEIGHTS
There was an slight increase in group mean liver weight values in rats receiving alkenes, C11/C12, hydroformylation products, distillation residues, compared to control values (Day 91/92); this increase only reached statistical significance in females given 1000 mg/kg/day and did not show a dose-related trend.

Absolute Organ Weights (g): Group Mean Values: Main Study

Group : 1 2 3 4
Test Item : Control ---- Test item ----
Dosage (mg/kg/day) : 0 100 300 1000

See Table 1 "Main Study" in "Any other information on results incl. tables"

There were no other test item-related organ weight changes. Occasional organ weight differences between treated and control groups were considered incidental; as there were no patterns, trends or correlating data to suggest these differences were toxicologically relevant, they were considered unrelated to administration of alkenes, C11/C12, hydroformylation products, distillation residues.

Test item-related liver weight changes noted at the terminal euthanasia were not observed at the end of the recovery period (Day 119). Occasional organ weight differences between treated and control groups were considered incidental; as there were no patterns, trends or correlating data to suggest these differences were toxicologically relevant, they were considered unrelated to administration of alkenes, C11/C12, hydroformylation products,
distillation residues.

See Table 2 "Recovery Study" in "Any other information on results incl. tables"

GROSS PATHOLOGY
No test item-related gross findings were noted among either main study or recovery animals. The gross findings observed were considered to be incidental, of the nature commonly observed in this strain and age of rats, and/or were of similar incidence in control and treated animals and, therefore, were considered to be unrelated to administration of the test item.

HISTOPATHOLOGY: NON-NEOPLASTIC
No test item-related microscopic findings were noted among either main study or recovery animals. The microscopic findings observed were considered to be incidental, of the nature commonly observed in this strain and age of rats, and/or were of similar incidence and severity in control and treated animals and, therefore, were considered to be unrelated to administration of the test item.

OTHER FINDINGS
None

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
organ weights and organ / body weight ratios
other: All parameters studied.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 1: Absolute Organ Weights (g): Group Mean Values: "Main Study"         
                                     
Group / Sex   Body Weight (g) Adrenals Brain Heart Kidneys Liver Lung Ovaries Pituitary Spleen
1F Mean 232 0.0695 1.98 0.81 1.47 7.87 1.23 0.099 0.012 0.47
  SD 23 0.0099 0.07 0.08 0.14 0.84 0.15 0.014 0.002 0.1
  n 9 9 9 9 9 9 9 9 9 9
                       
2F Mean 233 0.0759 1.97 0.84 1.51 8.21 1.22 0.096 0.013 0.42
  SD 14 0.0102 0.07 0.05 0.12 0.71 0.13 0.015 0.002 0.08
  n 10 10 10 10 10 10 10 10 10 10
                       
3F Mean 237 0.0698 2 0.84 1.54 8.12 1.26 0.103 0.012 0.46
  SD 14 0.006 0.07 0.08 0.11 0.69 0.19 0.019 0.002 0.04
  n 10 10 10 10 10 10 10 10 10 10
                       
4F Mean 231 0.0745 2 0.8 1.54 8.6 1.19 0.103 0.013 0.44
  SD 15 0.0124 0.08 0.07 0.16 0.51 0.13 0.015 0.002 0.06
  n 10 9 10 10 10 10 10 10 10 10
Significantly different from Group 1: a=p<0.05, b=p<0.01, c=p<0.001                     
 Table 2: Absolute Organ Weights (g): Group Mean Values: "Recovery Study"
              Group / Sex   Body Weight (g) Adrenals Brain Heart Kidneys Liver Lung Ovaries Pituitary Spleen
1F Mean 251 0.0777 2.02 0.95 1.61 7.52 1.26 0.111 0.015 0.46
  SD 9 0.0114 0.05 0.12 0.11 0.89 0.08 0.014 0.004 0.07
  n 5 5 5 5 5 5 5 5 5 5
                       
4F Mean 253 0.0824 2.07 0.92 1.66 7.68 1.35 0.114 0.014 0.47
  SD 26 0.0099 0.07 0.1 0.11 0.96 0.29 0.01 0.004 0.11
  n 5 5 5 5 5 5 5 5 5 5
Significantly different from Group 1: a=p<0.05, b=p<0.01, c=p<0.001                     

Applicant's summary and conclusion

Conclusions:
Administration of Alkenes, C11/C12, hydroformylation products, distillation
residues by once daily oral gavage was well tolerated in rats at levels of 100, 300 and
1000 mg/kg/day. Based on the results, the no-observed-effect level (NOEL) for males was
considered to be 1000 mg/kg/day. Due to reversible changes in the liver weights in females
the no-observed-adverse-effect level (NOAEL) was considered to be 1000 mg/kg/day.
Executive summary:

The objective of this study was to determine the potential toxicity of Alkenes, C11/C12, hydroformylation products, distillation residues when given by oral gavage for 90 or 91 days to rats and to evaluate the potential reversibility of any findings.

The following parameters and end points were evaluated in this study: clinical signs, body weights, body weight changes, food consumption, ophthalmology, detailed functional observationsclinical pathology parameters (haematology, coagulation, clinical chemistry, and urinalysis), gross necropsy findings, organ weights, and histopathologic examinations.

There was one unscheduled death during the study that was not related to administration of the test item. There were no clinical signs that were considered to be attributable to treatment and there were no test item-related changes in body weight or food consumption.

There were no ophthalmoscopic findings that could be attributed to treatment.

There were no differences in clinical signs seen during the functional observation battery recordings considered to be related to treatment.

There were no differences in haematology, coagulation, clinical chemistry or urinalysis parameters that were considered to be related to treatment.

In organ weights, liver weight was slightly increases in females at1000 mg/kg/day at the end of treatment period, with no correlating gross, microscopic or clinical chemistry correlates.

Following a 28 day recovery period, this increase was no longer observed, indicating reversibility.

No test item-related macroscopic or microscopic findings were noted among either main study or recovery animals.

In conclusion, administration of Alkenes, C11/C12, hydroformylation products, distillation residues by once daily oral gavage was well tolerated in rats at levels of 100, 300 and 1000 mg/kg/day. Based on the results, the no-observed-effect level (NOEL) for males was considered to be 1000 mg/kg/day. Due to reversible changes in the liver weights in females the no-observed-adverse-effect level (NOAEL) was considered to be 1000 mg/kg/day.