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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data from peer reviewed journal

Data source

Reference
Reference Type:
publication
Title:
Sperm function and fertility profile of test material in male rats
Author:
P. Oyeyipo et al.
Year:
2014
Bibliographic source:
International Journal of Green Pharmacy, 2014

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Reproductive toxicity study of test material was performed on male albino rats.
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Nicotine dihydrogen tartrate
- Molecular formula: C10H14N2.2C4H6O6
- Molecular weight: 462.4054 g/mol
- Substance type: Organic
- Physical state: No data

Test animals

Species:
rat
Strain:
other: albino rats
Details on species / strain selection:
No data available
Sex:
male
Details on test animals and environmental conditions:
Details on test animals and env. conditions
TEST ANIMALS
- Source: Animal House, College of Medicine,
University of Ibadan, Oyo State, Nigeria
- Age at study initiation: 8-10 weeks old
- Weight at study initiation: average weight ranged between 150 and 180 g
- Fasting period before study:
- Housing:
- Use of restrainers for preventing ingestion (if dermal):
yes/no
- Diet (e.g. ad libitum): ad libitum access to rat chow food
- Water (e.g. ad libitum): ad libitum access to drinking water.
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12:12‑hour light–dark


Administration / exposure

Route of administration:
oral: gavage
Vehicle:
physiological saline
Details on exposure:
Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in normal saline
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0, 1.0 mg/kg bw/day
- Amount of vehicle (if gavage): No data available

- Lot/batch no. (if required): No data available
- Purity: No data available

Other: The working solutions were stored in foil‑wrapped glass bottle at 4°C for no longer than 10 days
Details on mating procedure:
- M/F ratio per cage:4:5
- Length of cohabitation: 5days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:The presence of a vaginal plug was accepted as the index for a positive mate and taken as day one of pregnancy
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- After successful mating each pregnant female was caged (how):
- Any other deviations from standard protocol:
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
30 days
Frequency of treatment:
Daily
Details on study schedule:
No data available
Doses / concentrations
Remarks:
0, 1.0mg/kg bw/day
No. of animals per sex per dose:
32 male rats
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available

Examinations

Parental animals: Observations and examinations:
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: daily


BODY WEIGHT: Yes
Time schedule for examinations: daily
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.

Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day:
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
Time schedule for examinations:

OTHER:
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
Parameters examined in {P} male parental generations:
testis weight, epididymis weight, daily sperm production, sperm count in testes, sperm count in epididymides, , sperm motility, sperm morphology, were observed
Litter observations:
The number of litters delivered and their body weights were determined.
Postmortem examinations (parental animals):
No data available
Postmortem examinations (offspring):
No data available
Statistics:
Data obtained were expressed in Mean ± SEM. Statistical analysis was performed by analysis of variance (ANOVA) followed by multiple comparison by two‑tailed t‑test. The values for P < 0.05 were considered to be statistically significant.
Reproductive indices:
No data available
Offspring viability indices:
No data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
effects observed, treatment-related
Description (incidence and severity):
Oral administration of 1.0 mg/kg BW of test material on animals daily for a period of 4 weeks significantly
decrease (P < 0.05) the progressive motility of the sperm when compared with the control group

The mean epididymal sperm count of animals administered with 1.0 mg/kg BW was significantly decreased (P < 0.05) when compared with their control

A significant decrease (P > 0.05) was recorded for the mean percentage live sperm of rats treated with 1.0 mg/k bw when compared with the control

The test material caused an insignificant decrease (P > 0.05) in the epididymal sperm volume in the treated groups when compared with the control

Administration of 1.0 mg/kg bw test material significantly reduced normal sperm morphology

The mean serum testosterone level of animals that received 1.0 mg/kg bw of nicotine was significantly decreased (P < 0.05) when compared with the control group.
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
Administration of 1.0 mg/kg bw test material caused significant decrease in libido score when compared with the control, test material significantly decreased percentage fertility when compared with the control

Effect levels (P0)

Dose descriptor:
LOAEL
Effect level:
1 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
reproductive function (sperm measures)
reproductive performance
Remarks on result:
other: effects observed treatment related

Target system / organ toxicity (P0)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Effect levels (F1)

Dose descriptor:
other: not specified
Generation:
other: not specified
Based on:
test mat.
Sex:
not specified
Basis for effect level:
other: not specified
Remarks on result:
not measured/tested

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified

Any other information on results incl. tables

Semen parameters of experimental rats treated with test material

 

Dose

Motility (%)

Live/dead (%)

Volume (ml)

Count (106/ml)

ratio

 

 

 

 

Control

85.50±3.21

94.56±4.21

5.22±0.04

112.40±10.40

Nicotine

 

 

 

 

1.0 mg/kg+

32.00±4.65*

79.80±5.17*

5.18±0.06

64.20±5.60*

Values are expressed as means ± S.E.M of 8 rats per group. *P<0.05 vs control

 

Fertility profile of experimental rats treated with test material

Dose

Libido

score

Litter

number

Litter

weight (g)

Percentage

fertility

Control

8.82 ± 1.21

6.24 ±040

5.96± 0.20

100

1.0 mg/kg+ test material

3.26 ±1.62*

0.00 ±0.00*

0.00 ±0.00*

0*

 

Values are expressed as means ± S.E.M of 8 rats per group. *P< 0.05 vs control

 

Applicant's summary and conclusion

Conclusions:
Low Observed Adverse Effect Level (LOAEL) was considered to be 1.0 mg/kg/day, When male albino rats were treated with test material orally.
Executive summary:

Sperm function and fertility profile of test material was performed on 32 male albino rats whose average weight ranged between 150 and 180 g (8-10 weeks old). The test material dosage freshly prepared in normal saline for each group of animals was delivered orally at 1.0 mg/kg body weight (BW). The working solutions were stored in foilwrapped glass bottle at 4°C for no longer than10 days. A total of 20 untreated fertile, Proestrum female rats were used for the fertility test. Five untreated female rats were cohabited with each other of the four male groups from the day 31 of treatment. All animals were cohabited for 5 days. The presence of a vaginal plug was accepted as the index for a positive mate and taken as day one of pregnancy. The number of litters delivered and their body weights were determined. Oral administration of 1.0 mg/kg BW of test material on animals daily for a period of 4 weeks significantly decrease (P< 0.05) the progressive motility of the sperm when compared with the control group. The mean epididymal sperm count of animals administered with 1.0 mg/kg BW was significantly decreased (P< 0.05) when compared with their control. A significant decrease (P> 0.05) was recorded for the mean percentage live sperm of rats treated with 1.0 mg/k bw when compared with the control. The test material caused an insignificant decrease (P> 0.05) in the epididymal sperm volume in the treated groups when compared with the control. A significant decrease (P> 0.05) was recorded for the mean percentage live sperm of rats treated with 1.0 mg/k bw when compared with the control. The test material caused an insignificant decrease (P> 0.05) in the epididymal sperm volume in the treated groups when compared with the control. Administration of 1.0 mg/kg bw test material significantly reduced normal sperm morphology. The mean serum testosterone level of animals that received 1.0 mg/kg bw of nicotine was significantly decreased (P< 0.05) when compared with the control group.