Nicotine dihydrogen ditartrate

Toxicity to reproduction

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

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

Test material information

Constituent 1

Reference substance name:

Nicotine dihydrogen ditartrate

EC Number:

200-607-2

EC Name:

Nicotine dihydrogen ditartrate

Cas Number:

65-31-6

Molecular formula:

C10-H14-N2.2C4-H6-O6

IUPAC Name:

nicotine dihydrogen ditartrate

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 or test system 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 (10^6/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 foil‑wrapped 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.