Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
6 November 2007 - 27 August 2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: compliant to GLP and testing guideline
Cross-reference
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Except for absence of chemical analysis of the dosage forms
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): LCE07051
- Substance type: UVCB
- Physical state: white powder
- Analytical purity: not indicated
- Impurities (identity and concentrations): not indicated
- Composition of test material, percentage of components: not indicated
- Purity test date: not indicated
- Lot/batch No.: T70615
- Expiration date of the lot/batch: 7 February 2009
- Stability under test conditions: not indicated
- Storage condition of test material: at room temperature, protected from sunlight

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: breeder
- Age at first treatment: approx. 7 weeks
- Weight at first treatment (mean): M=304 g, F=199 g
- Housing: individually
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Relative humidity: 50 ± 20%
- Light/dark cycle: 12h/12h
- Ventilation: 12 cycles/hour of filtered, non-recycled air.

IN-LIFE DATES: beginning: 19 November 2007 / end: 20 December 2007

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
test item heated to 80°C, mixed with vehicle heated to 80°C, forming a solution.
The test item dosage forms were prepared daily

VEHICLE
- Justification for use and choice of vehicle (if other than water): lipophilicity of the substance
- Concentration in vehicle: 10, 30 and 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg/day
- Lot/batch no. (if required): 057K6093
- Purity: not indicated
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Absence of a practical method of analysis
Duration of treatment / exposure:
28 days
Frequency of treatment:
once daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 100, 300, 1000 mg/kg/day (m/f)
Basis:
other: nominal per gavage
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: no serious effects at up to 1000 mg/kg/day in a 7-day range-finding study, see 7.5.1
- Rationale for animal assignment: computerized stratification procedure (average body weight of each group is similar)
- Post-exposure recovery period, satellite groups: not performed
Positive control:
None

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule: day 1 and then weekly

FOOD CONSUMPTION for each animal: Yes
- Time schedule: weekly (last 7-day consumption period)

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY and CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: day 23 to 26
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: all

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule: day 22
- Dose groups that were examined: all
- Battery of functions tested: behavior / reflexes / sensory activity / grip strength / motor activity / rectal temperature

REPRODUCTION/FERTILITY: Yes
- Vaginal smears for oestrous cycles: each morning from day 22
- Seminology: at sacrifice, on all males : epididymal sperm motility and morphology ; epididymal and testicular sperm counts
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, all animals
ORGAN WEIGHTS: Yes, all animals
HISTOPATHOLOGY: Yes, all animals

all : see Table 1

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:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL CHEMISTRY
(Study director's opinion, see CSR author's opinion under "Effect levels") blood biochemistry analysis revealed higher triglycerides levels for male and female groups treated at 300 or 1000 mg/kg/day and higher urea and cholesterol levels for male and female groups treated at 1000 mg/kg/day.

SEMINOLOGY:
Epididymal sperm count and the percentage of morphologically normal sperm were lower for all groups treated with the test item and a slight effect was also observed on testicular sperm count. However since there were no treatment-related macroscopic or microscopic findings, and no dose-relationship, this was not considered unlikely to be test item-related by the study pathologist.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Remarks:
set by study director
Effect level:
ca. 100 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Dose descriptor:
NOAEL
Remarks:
revised by CSR authors
Effect level:
ca. 1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 2: Effects at blood biochemistry

Sex

Male

Female

Dose-level (mg/kg/day)

0

100

300

1000

0

100

300

1000

Urea (mmol/L)

2.7

3.2

3.1

3.5*

3.0

3.2

3.1

3.3

Cholesterol (mmol/L)

1.8

1.9

2.0

2.1*

1.5

1.6

1.5

1.9*

Triglycerides (mmol/L)

1.41

1.44

1.68

1.50

0.49

0.47

0.54

0.61

*: p0.05

Table 3: Quantitative effects on sperm

Dose-level (mg/kg/day)

100 

300

1000
 Number of spermatozoa in cauda epididymis (M)  120.0 81.5 80.2 85.4
 Number of spermatozoa per g of cauda of epididymis (M/g)  441 346  319 375
 Percent motile epididymal sperm (%)  96.7  93.3 91.6 92.3
 Number of sperm heads per g of testis (M/g)  126.7 120.3 116.8 114.7

Table 4: Qualitative effects on sperm    

Dose-level (mg/kg/day)

100

300

1000
 Normal (%) 93.8 86.5 86.6 90.6
 Normally shaped head separated from flagellum (%)  3.0 9.5  7.4 3.6

Applicant's summary and conclusion

Conclusions:
Under the experimental conditions of this study, the No Observed Adverse Effect Level (NOAEL) was considered by the study director to be 100 mg/kg/day based on the blood biochemistry effects. The CSR authors upgraded it to 1000 mg/kg/day as the biochemical effects were either negligible and/or related to the expectable metabolism of the components of the UVCB (see CSR section 5.1: fatty acid metabolism).
Executive summary:

The test item was administered once daily by gavage to rats at the dose-levels of 100, 300 or 1000 mg/kg/day for 4 weeks.

No treatment-related effects were observed on mean body weight gain, food consumption, Funtional Observation Battery or hematology and no adverse clinical signs were recorded. There were no effects on female estrous cycles.

Cholesterol and triglycerides levels were increased in male and female groups treated at 300 and 1000 mg/kg/day and urea levels were also higher at 1000 mg/kg/day. These effects, being observed in both males and females, were considered to be treatment-related and adverse (Study director's opinion, see conclusion of CSR authors above).

Sperm analysis of the left testis and epididymis revealed a decreased epididymal sperm count and a lower percentage of morphologically normal sperm at all dose-levels. This could indicate an effect on epididymal maturation or testicular-epididymal transport although no effects were observed at microscopic examination of the right testis and epididymis and no effects were

observed on the organ weights. Particularly noticable in the epididymal sperm count was the homogeneity of the results among the males of each group and the fact that all the test item-treated males were affected. However the pathologist concluded : in view of the lack of relationship to dose, and the high incidence of unilateral spontaneous lesions in the testis and epididymis in this strain of rats, the decrease in sperm numbers was considered unlikely to be treatment-related.