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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Range finding study to GLP; well documented study report which meets basic scientific principles.

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1985
Report Date:
1985
Reference Type:
secondary source
Title:
Unnamed
Year:
2008

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Preliminary Reproduction Toxicity Screening Test (Precursor Protocol of GL 421)
Deviations:
yes
Remarks:
Small number of tested animals and limited number of tissues examined at necropsy
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Cereclor S52
- Substance type: technical product
- Physical state: clear, slightly viscous liquid
- Analytical purity: no data
- Impurities (identity and concentrations): No stabiliser
- Composition of test material, percentage of components: C14-17 chlorinated paraffin (51.8% chlorination)
- Lot/batch no: 306

Test animals

Species:
rat
Strain:
other: Charles River COBS CD
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: Charles River Breeding Labs. Portage, Michigan, USA- Age at study initiation: 57 days at receipt; 83 days at initiation of treatment (F0 males and females); 21 days for F1 males and females- Weight at study initiation: F0 females at day 0 gestation- 255-312 g- Fasting period before study: no- Housing: suspended wire-mesh cages or plastic breeding cages.- Food (eg ad libitum): ad libitum- Water (e.g. ad libitum): ad libitum- Acclimation period: 12 daysENVIRONMENTAL CONDITIONS- Temperature (°C): 18-24- Humidity (%): 20-78- Air changes (per hr): no data- Photoperiod (hrs dark / hrs light): 12/12IN-LIFE DATES: From: 1984-02-28 To: 1984-07-26

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
DIET PREPARATION- Rate of preparation of diet (frequency): Weekly- Mixing appropriate amounts with (Type of food): yes- Purina Certified Rodent Chow 5002- Storage temperature of food: no data
Details on mating procedure:
- M/F ratio per cage: 1 male/2 females- Length of cohabitation: up to 10 days- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy/gestation- After 10 days of unsuccessful mating, the female was rested for 3 days and then returned to a cage with two males- Further matings after two unsuccessful attempts: no - After successful mating each pregnant female was caged individually in plastic cage with wood chip bedding.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Diet samples were extracted (soxhlet, 12 h) with hexane. The hexane extract was evaporated to dryness, ignited in an oxygen atmosphere, and the hydrochloric acid produced absorbed into sodium hydroxide solution. The solution was titrated potentiometrically to determine chlorine concentration.
Duration of treatment / exposure:
F0 (parental) generation : 28 days prior to mating, during mating and (females only) up to post-natal day 21.F1 generation : throughout gestation, lactation and up to 70 days of age
Frequency of treatment:
daily in the diet
Details on study schedule:
F1 generation sacrificed on lactation days 6 or 21 or at 70 days of age
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:0, 100, 1000 and 6250 ppmBasis:nominal in diet
Remarks:
Doses / Concentrations:0, 6, 62 and 384 mg/kg bw/day for malesBasis:nominal in dietaverage dose over 11 weeks
Remarks:
Doses / Concentrations:0, 8, 74 and 463 mg/kg bw/day for femalesBasis:nominal in dietaverage dose
No. of animals per sex per dose:
5 males and 10 females (F0 and F1 generation)
Control animals:
yes, plain diet
Details on study design:
no data
Positive control:
no

Examinations

Parental animals: Observations and examinations:
Clinical observations, body weights, food consumption
Oestrous cyclicity (parental animals):
no data
Sperm parameters (parental animals):
no data
Litter observations:
Pup survival, physical and behavioural abnormalities
Postmortem examinations (parental animals):
Necropsy examination performed on kidneys, lungs, ureter and urinary bladder of F0 females only
Postmortem examinations (offspring):
Necrosy examination for gross abnormalities.
Statistics:
Fertility indices were compared using Chi-squared test or Fisher's exact test. Pup survival indices were compared by the Mann-Whitney U-test. Mean pup body weights and parental body weights and food consumption were compared by analysis of variance, Bartlett's test and the appropriate t-test.
Reproductive indices:
Fertility indices, copulatory interval, parturition, gestation length
Offspring viability indices:
Numbers of viable and stillborn pups, pup survival at birth, during lactation and at weaning

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:
not specified
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed

Reproductive function / performance (P0)

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

Details on results (P0)

No deaths in males or females and no histological abnormalities in females. Significant reduction in food intake for females at 6250 ppm diet during week 5. No treatment-related efefcts on fertility indices and no abnormalities noted in the histopathological examination of females.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
ca. 400 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects on fertility; C14-17 chlorinated paraffin (52% chlorination)

Results: F1 generation

General toxicity (F1)

Clinical signs:
effects observed, treatment-related
Mortality / viability:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, treatment-related
Histopathological findings:
not examined

Details on results (F1)

VIABILITY (OFFSPRING). At birth, F1 pup survival in all dose groups was equivalent to that of the control group F1 pups. However, there was a marked and statistically significant decrease in pup survival during lactation at 6250 ppm, such that none of the pups survived until weaning. Reduced pup survival (by 11%) during lactation was also evident at 1000 ppm and, although not statistically significant, is considered to be of toxicological importance.BODY WEIGHT (OFFSPRING) Dose-related, but not statistically significant, reductions in body weight gain occurred in the F1 pups in the mid- and high-dose groups relative to the control values on lactation days 7, 14 and 21, and in the low and mid-dose F1 groups post-weaning. The investigators regard these differences as being of "uncertain toxicological significance". CLINICAL SIGNS (OFFSPRING). Decreased activity and swollen and dark or black eye(s) were observed in a few F1 pups in 1 or 2 mid- and high-dose litters. Haematological analyses revealed “reductions” in erythrocyte counts, haemoglobin concentration and haematocrit among a single litter of F1 pups at the top dose on lactation day 6, compared to the controls on lactation day 7.GROSS PATHOLOGY. Increases in the occurrence and severity of subcutaneous haematoma, pallor, blood around the orifices, pale liver, kidneys, lungs and spleen and blood in the cranial cavity and brain were seen in F1 pups at 1000 and 6250 ppm. Haematoma was noted in all of the litters at 6250 ppm.OTHER FINDINGS (OFFSPRING) no further data

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
ca. 8 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Survival and necropsy finding in the pups (indications of internal haemorrhaging seen in mid- and high-dose groups); C14-17 chlorinated paraffin (52% chlorination)

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The administration of Cereclor S52 (a C14-17 chlorinated paraffin; 52% chlorination) to male and female rats in the diet for 28 days prior to pairing, and during mating and (in females only) gestation at up to approximately 400 mg/kg bw/day (the highest tested dose) had no effect upon fertility. The dams were also fed Cereclor S52 in the diet throughout lactation. Internal haemorrhaging was reported in some pups during the lactational period from dams administered about 74 mg/kg bw/day, with no pups from the top-dose group (dams receiving about 460 mg/kg bw/day during lactation) surviving to weaning. No treatment-related adverse effects were seen in the pups from the low-dose group (8 mg/kg bw/day).
Executive summary:

In a range-finding study, groups of 5 males and 10 female Charles River COBS CD rats were administered Cereclor S52 (a C14-17 chlorinated paraffin; 52% chlorination) in the diet at concentrations of 0, 100, 1000 and 6250 ppm for 28 days prior to mating, during mating and (in females only) up to lactational day 21. The average doses of test substance received were 0, 6, 62 or 384 mg/kg bw/day for males and 0, 8, 74 or 463 mg/kg bw/day for females. Five male and ten female pups were selected at random from each group, and fed the same diet as their parents from weaning up to 70 days of age. One litter from each group was killed on lactation days 6 (high dose) and 7 (controls); the remaining pups were sacrificed on lactation day 21. Necropsy examination was performed on kidneys, lungs, ureter, and urinary bladder only of F0 females but not males. A number of reproductive and litter parameter assessments were conducted following sacrifice. Blood samples were collected for haematological analysis from F1 pups at various timepoints.

No deaths occurred in the parental generation (F0), and there were no abnormalities noted in the histology examinations of F0 females. A statistically significant decrease (by 12%) in food consumption was seen in the high-dose females during week 5. No treatment-related effects on fertility indices were observed.

At birth, pup survival in all dose groups was equivalent to that of the control group pups. However, a statistically significant decrease in pup survival was noted during lactation in the top-dose group (probably as a result of the internal haemorrhaging), with no pups surviving until weaning. Reduced pup survival (by 11%) was also evident during the lactational period in the mid-dose group and, although not statistically significant, is considered to be of toxicological importance. Dose-related, but not statistically significant, reductions in body weight gain occurred in the F1 pups in the mid- and high-dose groups relative to the control values on lactation days 7, 14 and 21, and in the low and mid-dose groups post-weaning. With the exception of the aformentioned reductions in F1 body weight gain, no adverse treatment-related effects were seen in the low-dose group. Decreased activity and swollen and dark or black eye(s) were observed in a few pups in one or two mid- and high-dose litters. Haematological analyses revealed reductions in erythrocyte counts, haemoglobin concentration and haematocrit among a single litter of pups at the top dose on lactation day 6 compared to the controls on lactation day 7. At necropsy, dose-related, but not statistically significant, increases in the occurrence and severity of subcutaneous haematoma, pallor, blood around the orifices, pale liver, kidneys, lungs and spleen and blood in the cranial cavity and brain was seen in mid- and high-dose pups. Haematoma was noted in all of the high dose litters.

In this range-finding study, no adverse effects were seen on fertility indices following the administration of Cereclor S52 to rats in the diet at up to approximately 400 mg/kg bw/day. Significant effects were seen in the developing offspring prior to them having been weaned in the 1000 and 6250 ppm groups, suggesting a no-observed-adverse-effect level (NOAEL) in the offspring of approximately 8 mg/kg bw/day (as a maternal dose) for effects seen during the lactational period.