Registration Dossier

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
October 25 - December 20, 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
Qualifier:
according to
Guideline:
other: EPA OPPTS 870.3550 (Reproduction/Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Corsair Clear #34
- Substance type: Clear white lumpy solid
- Physical state: Solid
- Stability under test conditions: Stable
- Storage condition of test material: At room temperature in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS Rat: Crl:WI(Han)
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: (P) Approximately 10 weeks.
- Weight at study initiation: (P) Males: 294-322 g; Females: 212-242 g;
- Fasting period before study: not applicable
- Housing:
Pre-mating: Animals were housed in groups of 5 animals/sex/cage in Macrolon cages (MIV type).
Mating: Females were caged together with males on a one-to-one-basis in Macrolon cages (MIII type).
Post-mating: Males were housed in their home cage (Macrolon cages, MIV type) with a maximum of 5 animals/sex/cage. Females were individually housed in Macrolon cages (MIII type).
Lactation: Offspring was kept with the dam until termination.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 5 days prior to start of treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.1 – 22.8°C
- Humidity (%): 25 - 95%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Formulations (w/w) were prepared daily within 4 hours prior to dosing and were homogenised to a visually acceptable level. Adjustment was made for specific gravity of the vehicle. No correction was made for the purity of the test substance.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed at NOTOX.
- Concentration in vehicle: 10, 30, 200 mg/ml
- Amount of vehicle (if gavage): 5 mL/kg body weight
Details on mating procedure:
- M/F ratio per cage: 1/1
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of post-coitum.
- After successful mating each pregnant female was caged individually.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analyses were done a single occasion during the treatment phase (16 November 2007) according to a validated HPLC-UV method (see section 8) :
group 1 (vehicle control): accuracy (middle position of container)
group 2 (50 mg/kg bw/d): accuracy and homogeneity (top/middle/bottom position of container)
group 3 (150 mg/kg bw/d): accuracy (middle position of container)
group 4 (1000 mg/kg bw/d): accuracy and homogeneity (top/middle/bottom position of container)
The concentrations analysed in the formulations of Group 2, Group 3 and Group 4 were in agreement with target concentrations (i.e. mean accuracies between 101% and 103%). No test substance was detected in the Group 1 formulations. The formulations of Group 2 and Group 4 were homogeneous (i.e. coefficient of variation ≤ 10%).
Duration of treatment / exposure:
Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to termination. Females were exposed for 42 to 53 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 3 days of lactation.
Frequency of treatment:
Once daily for 7 days per week, approximately the same time each day with a maximum of 4 hours difference between the earliest and latest dose. Animals were dosed up to the day prior to scheduled necropsy.
Details on study schedule:
- Age at mating of the mated animals in the study: 12 weeks
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
1000 mg/kg bw/d
Basis:
actual ingested
Remarks:
Doses / Concentrations:
150 mg/kg bw/d
Basis:
actual ingested
Remarks:
Doses / Concentrations:
50 mg/kg bw/d
Basis:
actual ingested
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were based on a 5-day dose range finding study in which female Wistar Han rats were exposed to 500 and 1000 mg/kg for five consecutive days. No toxicity was observed.

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least twice daily.
- Cage side observations : Mortality / viability.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least once daily

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on days 0, 4, 7, 11, 14, 17 and 20 of gestation, and during lactation on days 1 and 4.

FOOD CONSUMPTION : Yes, weekly.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

WATER CONSUMPTION : No

REPRODUCTION PROCESSES: Male number paired with, mating date, confirmation of pregnancy, and delivery day were recorded.
Sperm parameters (parental animals):
Of the males of the control and high dose group, additional slides of the testes were prepared to examine staging of spermatogenesis.
Litter observations:
Mortality / Viability: The numbers of live and dead pups at the First Litter Check (= check at day 1 of lactation) and daily thereafter were determined. If possible, defects or cause of death were evaluated.
Clinical signs: At least once daily, detailed clinical observations were made in all animals.
Body weights: Live pups were weighed during lactation on days 1 and 4.
Sex: Was determined for all pups on days 1 and 4 of lactation (by assessment of the ano-genital distance).
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals, following completion of the mating period (a minimum of 28 days of dose administration.
- Maternal animals: All surviving animals, on lactation day 4 or shortly thereafter.
- Non-maternal animals: On post-coitum day 25-27 (females with evidence of mating) or approximately 21 days after the last day of the mating period (females without evidence of mating), the F0- females which had not delivered were sacrificed.

GROSS NECROPSY
- Gross necropsy consisted of macroscopic examination of the cranial, thoracic and abdominal tissues and organs, with special attention being paid to the reproductive organs.

HISTOPATHOLOGY / ORGAN WEIGHTS
Samples of the following tissues and organs were collected:
Cervix, Clitoral gland, Coagulation gland, Epididymides, Ovaries, Pituitary gland, Preputial gland, Prostate gland, Seminal vesicles, Testes, Uterus, Vagina, All gross lesions.
Epididymides and Testes were weighed.
Postmortem examinations (offspring):
Pups were killed by decapitation on day 4 of lactation or shortly thereafter. All offspring was sexed and externally examined if practically possible. The stomach was examined for the presence of milk. Descriptions of all external abnormalities were recorded. If possible, defects or cause of death were evaluated.
Statistics:
The following statistical methods were used to analyse the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (Dunnett, 1955) (many-to-one t-test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (Miller, 1981) (many-to-one rank test) was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test (Fisher, 1950) was applied to frequency data.
All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
No statistical analysis was performed on histopathology findings.
Reproductive indices:
Percentage mating: Number of females mated x 100/Number of females paired
Fertility index: Number of pregnant females x 100/Number of females paired
Conception rate: Number of pregnant females x 100/Number of females mated
Gestation index: Number of females bearing live pups x 100/Number of pregnant females
Duration of gestation: Number of days between confirmation of mating and the beginning of parturition
Offspring viability indices:
Percentage live males at First Litter Check: Number of live male pups at First Litter Check x 100/Number of live pups at First Litter Check
Percentage live females at First Litter Check: Number of live female pups at First Litter Check x 100/Number of live pups at First Litter Check
Percentage of postnatal loss days 0-4 post partum: Number of dead pups on day 4 post partum x 100/Number of live pups at First Litter Check
Viability index: Number of live pups on day 4 post partum x 100/Number of pups born alive

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed

Details on results (P0)

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
At 1000 mg/kg, males showed a slightly decreased body weight and body weight gain on Day 1 of the mating period (Day 15 of treatment). No body weight changes were observed for males up to 150 mg/kg and for females up to 1000 mg/kg.
At 1000 mg/kg, males showed a decreased absolute (statistical significant) and relative (not statistical significant) food consumption during Days 8-15 of treatment. At 150 mg/kg, a reduced food consumption was noted for females on Days 11-14 post-coitum. As this single occasion and as no dose response relationship was noted, this change was not considered toxicologically relevant.

Effect levels (P0)

Dose descriptor:
NOAEL
Remarks:
parental, reproduction, breeding
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: There were no treatment related changes for pup development up to 1000 mg/kg.

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Corsair Clear #34 was administered by daily oral gavage to male and female Wistar Han rats at dose levels of 0, 50, 150 and 1000 mg/kg/day. The males were exposed for 2 weeks prior to mating, during mating, and up to termination (for 29 days). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and at least 3 days of lactation (for 42 to 53 days).
Formulation analysis showed that the formulations were prepared accurately and were homogeneous.
At 1000 mg/kg, males showed a slightly decreased body weight and body weight gain on Day 15 of treatment and slightly decreased food consumption during Days 8-15 of treatment. As these findings were very slight and recovered during treatment, they were not considered toxicologically significant.
There were no treatment related changes for mortality, clinical signs, macroscopic and microscopic examination, organ weights, reproduction, breeding data and pup development up to 1000 mg/kg.
In conclusion, treatment with Corsair Clear #34 by oral gavage in male and female Wistar Han rats at dose levels of 50, 150 and 1000 mg/kg body weight/day revealed no parental, reproduction, breeding and developmental toxicity for treatment up to 1000 mg/kg body weight/day.
Based on these findings, the parental, reproduction, breeding and developmental No Observed Adverse Effect Level (NOAEL) was established at least at 1000 mg/kg body weight/day.