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Key value for chemical safety assessment

Effects on fertility

Description of key information

The NOAEL of Everlan SL65 in rat’s reproductive performance was 1000 mg/kg/day (OECD TG421).

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
June 16, 2016 - Jan 11, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Guideline study done to GLP.
Qualifier:
according to guideline
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Species:
rat
Strain:
other: Crl:CD (SD)
Sex:
male/female
Details on test animals or test system and environmental conditions:
- Source: BioLASCO Taiwan Co., Ltd., Taipei, Taiwan
- Age at study initiation: about 9-week old
- Weight at study initiation: Males: 330-383 g; Females: 211-253 g
- Housing: Except for the mating period, animals were housed individually in polycarbonate cages. While mating, animals were pair-housed in stainless steel wire mesh cages.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 11 days
- Temperature (°C): 21 ± 2 °C
- Humidity (%): 50 ± 20%
- Photoperiod: 12-hrs dark / 12-hrs light
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
Water for injection
Details on mating procedure:
- M/F ratio per cage: 1:1
- After 4 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- After successful mating each pregnant female was caged (how): 1 animal per cage
Duration of treatment / exposure:
For male: 28 days
For female: 40-53 days
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Dose Group 1 (Control)
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
Dose Group 2 (low dose)
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
Dose Group 3 (mid dose)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
Dose Group 4 (high dose)
No. of animals per sex per dose:
10 males and 10 females per dose
Control animals:
yes, concurrent vehicle
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
These observations included red-brown colored feces, soft feces, unformed feces, amber/red-brown/yellow colored urine, the red, red-brown, yellow or brown hair stains on the whole body and red hair stains on the nose in all groups.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
In males, statistically significantly reduced body weight gain compared to the controls was observed in Group 4 animals on Day8. The total body weight gain of Group 4 was also lower than the body weight gain of the control animals between D1 and D 29, but the difference did not reach statistical significance difference.
In females, reduced body weight gain was also observed reaching statistical significance compared to the controls in Group 4 on D15 and in Groups 3 and 4 on G6.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
effects observed, non-treatment-related
Description (incidence and severity):
Statistically significantly increased food consumption compared to the controls was observed in males of Groups 3 and 4 on D15, in females of Group 2 on D13 and in females of Group 4 on G6, G13, G20 and P4.
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not specified
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Discolouration of urine from Day 2 in high dose groupe rats.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
There were no statistically significant differences in the number of corpora lutea and implantations between control and treated groups. Pre-implantation and pre-natal losses were observed in all groups including the controls. The only treatment-related gross finding was discoloration of the cecum in 3 animals of Group 4. Decreased size of thymus was observed in one animal each of Groups 3 and 4
Reproductive function: oestrous cycle:
effects observed, non-treatment-related
Description (incidence and severity):
In males, a positive indication of mating was obtained for 9, 10, 9 and 8 of 10 animals in Groups 1 to 4, resulting in a mating index of 90%, 100%, 90% and 80%, respectively, indicative of a slight decrease at Group 4.
In females, a positive indication of mating was obtained for 10 of 10 animals in Groups 1, 2 and 3 and for 9 of 10 animals in Group 4. The mating index was 100%, 100%, 100% and 90% for Group 1 to 4, respectively.
There was one animal in Group 1 and 3, respectively, that showed a pre-coital interval longer 5 days. In each of these females, the mating with the first male failed but the copulation with another male was successful. In Groups 2 to 4, 1 or 2 animals with positive indications of mating was later found not to be pregnant. All Group 1 animals were found to be pregnant. Thus, the fertility index was 100%, 90%, 90% and 78% in the control, low, mid and high dose group, respectively, indicative of a slight decrease in Group 4. The average length of gestation was 21.7-22 days without dose-dependency.
Reproductive function: sperm measures:
not examined
Reproductive performance:
effects observed, non-treatment-related
Description (incidence and severity):
All pregnant rats delivered live pups. The gestation index was 100% in all groups. One and three dead pups were observed at birth in Groups 2 and 4. The incidence of dams with dead pups at birth was 0%, 11%, 0% and 29% in Group 1, 2, 3 and 4, respectively. The dead pup incidence in Group 4 was slightly higher than in other groups. However, the number of pups per litter at birth in Group 4 was higher than in other group. Moreover, the number of stillborns per litter was 0.42. On P4, a decreased number of live pups was observed in Groups 2 and 4. The 4-day survival index was 100.0%, 97.0%, 100% and 97.2% in Groups 1 to 4, respectively. No external abnormally pups were observed at birth in the control and treated groups. At birth, 45.4% to 54.2% of pups were male and on P4, 46.2% to 59.6% pups were male. The litter weights in all groups increased from P0 to P4.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food efficiency
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
reproductive function (oestrous cycle)
reproductive function (sperm measures)
reproductive performance
Critical effects observed:
no
Lowest effective dose / conc.:
1 000 mg/kg bw/day (nominal)
System:
male reproductive system
Organ:
testes
Critical effects observed:
no
Lowest effective dose / conc.:
1 000 mg/kg bw/day (nominal)
System:
female reproductive system
Organ:
uterus
Clinical signs:
not specified
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, non-treatment-related
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Dose descriptor:
other:
Remarks on result:
not measured/tested
Remarks:
OECD 421 method is not defined the does object for the pups.
Key result
Reproductive effects observed:
no
Lowest effective dose / conc.:
1 000 mg/kg bw/day (nominal)
Treatment related:
no

For male: DX mean DayX

For female: DX mean DayX during pre-mating; GX mean DayX during gestation; PX mean DayX during lactating

 

Dose Formulation Analysis

On the first and last preparation, the difference between the mean value and the targeted concentration for all dose group formulations were within ± 15.0%. The homogeneity verification revealed that the precision of samples for the low and high dose formulation were ≤ 10.0%.

 

Mortality and Clinical Observations

All animals survived the study period. These observations included red-brown colored feces, soft feces, unformed feces, amber/red-brown/yellow colored urine, partial hair loss, the red, red-brown, yellow or brown hair stains on the whole body and red hair stains on the nose in all groups. For only one male and one female rat at the high dose level, the salivation was observed for two days.

 

Body Weights

In males, statistically significantly reduced body weight gain compared to the controls was observed in Group 4 animals on Day8. The total body weight gain of Group 4 was also lower than the body weight gain of the control animals between D1 and D 29, but the difference did not reach statistical significance difference. In females, reduced body weight gain was also observed reaching statistical significance compared to the controls in Group 4 on D15 and in Groups 3 and 4 on G6.

 

Food Consumption

Statistically significantly increased food consumption compared to the controls was observed in males of Groups 3 and 4 on D15, in females of Group 2 on D13 and in females of Group 4 on G6, G13, G20 and P4.

 

Cohabitation and Pregnancy

In males, a positive indication of mating was obtained for 9, 10, 9 and 8 of 10 animals in Groups 1 to 4, resulting in a mating index of 90%, 100%, 90% and 80%, respectively, indicative of a slight decrease at Group 4. In females, a positive indication of mating was obtained for 10 of 10 animals in Groups 1, 2 and 3 and for 9 of 10 animals in Group 4. The mating index was 100%, 100%, 100% and 90% for Group 1 to 4, respectively. There was one animal in Group 1 and 3, respectively, that showed a pre-coital interval longer 5 days. In each of these females, the mating with the first male failed but the copulation with another male was successful. In Groups 2 to 4, 1 or 2 animals with positive indications of mating was later found not to be pregnant. All Group 1 animals were found to be pregnant. Thus, the fertility index was 100%, 90%, 90% and 78% in the control, low, mid and high dose group, respectively, indicative of a slight decrease in Group 4. The average length of gestation was 21.7-22 days without dose-dependency.

 

Litter Observation

All pregnant rats delivered live pups. The gestation index was 100% in all groups. One and three dead pups were observed at birth in Groups 2 and 4. The incidence of dams with dead pups at birth was 0%, 11%, 0% and 29% in Group 1, 2, 3 and 4, respectively. The dead pup incidence in Group 4 was slightly higher than in other groups. However, the number of pups per litter at birth in Group 4 was higher than in other group. Moreover, the number of stillborns per litter was 0.42. On P4, a decreased number of live pups was observed in Groups 2 and 4. The 4-day survival index was 100.0%, 97.0%, 100% and 97.2% in Groups 1 to 4, respectively. No external abnormally pups were observed at birth in the control and treated groups. At birth, 45.4% to 54.2% of pups were male and on P4, 46.2% to 59.6% pups were male. The litter weights in all groups increased from P0 to P4.

 

Male Gross Examination, Organ Weight and Histopathology

There were no statistically significant differences in testis and epididymis weights between control and treated groups. No gross changes were observed in any males.

 

Females Gross, Uterus Examination and Histopathology

There were no statistically significant differences in the number of corpora lutea and implantations between control and treated groups. Pre-implantation and pre-natal losses were observed in all groups including the controls. The only treatment-related gross finding was discoloration of the cecum in 3 animals of Group 4. Decreased size of thymus was observed in one animal each of Groups 3 and 4

Conclusions:
According to OECD 421 test method, the no observed adverse effect level of Everlan SL65 in rat's reproductive performance was 1000 mg/kg/day.
Executive summary:

This test using the procedures outlined in the QPS Study Plan for T65315036-RP and OECD 421. Everlan SL65 (100, 300 or 1000 mg/kg/day) was given orally by gavage once daily to male rats for 28 days and female rats for 40 to 53 days, depending on the time of copulation and gestation. Dose-related clinical observations in the treated groups included test article-like colored/soft/unformed feces or urine and hair stained. Reduced body weight gain and increased food consumption were observed in males and females at Group 3 or 4. The only treatment-related gross findings was discoloration of the cecum in three females at Group 4. However, there were no definitely test article-related effects on male and female rats’ reproductive performance such as gonadal function, mating behavior, conception, development of the conceptus and parturition. The no observed adverse effect level of Everlan SL65 in rat’s reproductive performance was 1000 mg/kg/day.

 

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Effect on fertility: via oral route

Everlan SL65 (100, 300 or 1000 mg/kg/day) was given orally by gavage once daily to male rats for 28 days and female rats for 40 to 53 days, depending on the time of copulation and gestation. Dose-related clinical observations in the treated groups included test article-like colored/soft/unformed feces or urine and hair stained. Reduced body weight gain and increased food consumption were observed in males and females at Group 3 or 4. The only treatment-related gross findings was discoloration of the cecum in three females at Group 4. However, there were no definitely test article-related effects on male and female rats’ reproductive performance such as gonadal function, mating behavior, conception, development of the conceptus and parturition. The no observed adverse effect level of Everlan SL65 in rat’s reproductive performance was 1000 mg/kg/day.

Effects on developmental toxicity

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available

Justification for classification or non-classification

Additional information