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Toxicological information

Toxicity to reproduction

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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:
2002-03-25 to 2002-11-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was carried out in accordance with an appropriate OECD test guideline and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Canada, 188 LaSalle, St. Constant, Canada
- Age at study initiation: >=8 wk
- Weight at study initiation: 187-238 g (f); 250-313 g (m)
- Housing: 1/suspended wire cage
- Diet: standard diet ad libitum
- Water: drinking water ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: From: 2002-03-25 To: 2002-11-07

Administration / exposure

Route of administration:
inhalation
Type of inhalation exposure (if applicable):
whole body
Vehicle:
unchanged (no vehicle)
Details on exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 450 l steel and glass Rochester-style inhalation chambers with stainless steel exposure caging.
- Method of holding animals in test chamber: individual stainless steel caged compartments
- Method of conditioning air: air passed through series of purification filters
- System of generating vapour/aerosols: Test substance placed in warming chamber (75 deg C). The liquid TS was then metered into a heated metal J-tube for vapourization.
- Air change rate: at 10 exchanges of chamber volume per h

TEST ATMOSPHERE
- Brief description of analytical method used: GC
- Samples taken from breathing zone: automatic sampling from chamber

Acclimatization for 3 h on 2 days
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: continuously until evidence of copulation
- Proof of pregnancy: vaginal plug or sperm in vaginal smear (day 0)

Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
Exposure period: 28 days for males and 46 days for females
Premating exposure period (males): 2 weeks
Premating exposure period (females): 2 weeks
Frequency of treatment:
6 hours/day; 7 days/week
Details on study schedule:
IUCLID4 Number of generation studies: 1
Doses / concentrations
Remarks:
Doses / Concentrations:
100, 500, and 2500 ppm (nominal); 99, 512 and 2492 ppm (actual)
Basis:

No. of animals per sex per dose:
10
Control animals:
other: yes, control group was exposed to filtered air
Positive control:
No

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily or twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at sacrifice
- Anaesthetic used for blood collection: Ketamine, HCL/Xylazine
- How many animals: all toxicity groups

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at sacrifice
- How many animals: all toxicity groups

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: prior to exposure and during wk 4
- Dose groups that were examined: all toxicity groups
- Battery of functions tested: FOB and motor activity examinations

Postmortem examinations (parental animals):
 Reproductive/developmental phase females were not subjected to macroscopic examination at necropsy, however the number of corpora lutea and implantation sites was recorded. No microscopic evaluation was conducted on organs and tissues of reproductive/developmental phase females.
Toxicity group males and females: necropsy and microscopic examination of a wide range of tissues.
Postmortem examinations (offspring):
Pups found dead and pups sacrificed on day four postpartum were examined for external gross abnormalities only. No pup tissues were collected and the carcasses were discarded.
 
Statistics:
Statistical methods:'s test and Kolmogorov-Smirnov test. Parametric data was tested using one-way Analysis of Variance (ANOVA) followed by Dunnett's Test (if significant); nonparametric data was tested by Kruskal-Wallis Test followed by Wilcoxon. Categorical data and histomorphology findings were evaluated with Fisher's Exact Test. Statistically significant probabilities are reported at either the p<0.05 or p<0.01 levels. 

Results and discussion

Results: P0 (first parental animals)

Details on results (P0)

2500 ppm Females (Reproductive/Developmental Phase): Decreased litter size (33%*); Decreased litter weight (27%*); Decreased number of implantation sites (33%*) Males exposed to 2500 ppm had decreased weight of seminal vesicles and atrophy.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Effect level:
500 ppm
Basis for effect level:
other: Approx 4.55 mg/l
Dose descriptor:
LOAEC
Effect level:
2 500 ppm
Sex:
male/female
Basis for effect level:
other: Approx 22.8 mg/l. Male: atrophy of seminal vesicles. Female: reduced body weight

Results: F1 generation

Effect levels (F1)

open allclose all
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
500 ppm
Basis for effect level:
other: Approx 4.55 mg/l
Dose descriptor:
LOAEC
Generation:
F1
Effect level:
2 500 ppm
Sex:
not specified
Basis for effect level:
other: Approx 22.8 mg/l. Reduced litter size and number of inplantation sites.

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

In the reproductive/developmental phase of the study exposure to 2500 ppm D3 was associated with decreased litter size, litter weight and number of implantation sites. A NOAEC of 500 ppm (4.5 mg/L) and LOAEC of 2500 ppm (22.8 mg/L) for reproductive/developmental toxicity were determined based on the decreased litter size and implantation sites. The maternal NOAEC and LOAEC were 500 and 2500 ppm, respectively based on the decrease in body weights.

NOAEL (NOAEC):

Reproductive/Developmental phase: Decreased Litter Size/Implantation Sites: NOAEL = 500 ppm

 

Toxic Response/Effects by Dose Level:

2500 ppm
Females (Reproductive/Developmental Phase): Decreased litter size (33%*); Decreased litter weight (27%*); Decreased number of implantation sites (33%*)
Males exposed to 2500 ppm had decreased weight of seminal vesicles and
 atrophy.

 

Statistical Results, As Appropriate: "*" denotes statistically different from control (p<0.05).

The results can be summarized as follows:

Body weight: Reproductive/Developmental Phase Females: Decreased body weight (8%) on gestation day 20 at 2500 ppm

Food/water consumption: Reproductive/Developmental Phase Females: No treatment-related effect.

 

I. Description, severity, time of onset and duration of clinical signs:

Anogenital Staining: Reproductive/Developmental Phase: Present in 100 and 2500 ppm groups. Time of onset (first occurrence): study day 1. Duration (last noted occurrence): study day 34

Probable Porphyrin Staining: Reproductive/Developmental Phase: Present in 500 and 2500 ppm groups Time of onset (first occurrence): study day 4 Duration (last noted occurrence): study day 36. One isolated incident of a female rat in the 2500 ppm group was recorded where the rat was seen squealing and shaking with "bulging eyes" for 10 minutes during treatment on day 31.

 

II. Hematological findings incidence and severity: No treatment-related effects.

 

III. Mortality and time to death: All adult animals survived to their scheduled necropsy.  

 

IV. Gross pathology incidence and severity: No treatment-related macroscopic findings

 

V. Histopathology incidence and severity: Reproductive/Developmental Toxicity: Reduced litter size (33%), litter weight (27%), and number of implantation sites (33%) at 2500 ppm.

Applicant's summary and conclusion

Conclusions:
In a well reported combined repeated dose/reproductive and developmental inhalation toxicity screening test, exposure to 2500 ppm (22.8 mg/L) D3 was associated with decreased litter size, litter weight and number of implantation sites. The findings support a NOAEC of 500 ppm (4.5 mg/L). Maternal effects (reduced weight gain) were seen 2500 ppm.