Heparin, sodium salt

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Endpoint:

two-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: Limited study summary. Not GLP.

Qualifier:

no guideline followed

Principles of method if other than guideline:

The Sprague-Dawley rats were used in this fertility and reproductive study. The test substance was administered into dorsal region by subcutaneous route. 10 males and 20 females were treated 60 and 14 days, respectively, before mating with each dose of drug; furthermore females only received the treatment from the first day of pregnancy until the weaning of pups (day 23 after delivery).

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Female and male 8 weeks of age
- Weight at study initiation: Female and male weighing 200±10 g
- Housing: The animals were housed in Makrolon cages in an air-conditioned room.
- Diet (e.g. ad libitum): Females received pelletted dry diet Altromin MT and males diet Altromin R.
- Water (e.g. ad libitum): Tap water was available ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-23°C
- Humidity (%): 55-60%
- Air changes (per hr): Ventilation was 12 times/h
- Photoperiod (hrs dark / hrs light): Lighting schedule was 12:12 light/dark (from 7:00 a.m. to 7:00 p.m.)

No additional data

Route of administration:

subcutaneous

Vehicle:

physiological saline

Details on exposure:

The test substance dissolved in physiological solution and administered by s.c. route at 1, 5, 10 mg/kg/d at a volume of 5 mL/kg into the dorsal region.

Details on mating procedure:

- M/F ratio per cage: 1/3
- Length of cohabitation: one night
- Proof of pregnancy: sperm in vaginal smear referred to as day 1 of pregnancy

No additional data

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

10 males and 20 females were treated 60 and 14 days, respectively, before mating with each dose of drug; furthermore females only received the treatment from the first day of pregnancy until the weaning of pups (day 23 after delivery).

Frequency of treatment:

Once daily

Details on study schedule:

- F1 parental animals not mated until [10] weeks after selected from the F1 litters.

No additional data

Remarks:

Doses / Concentrations:
1, 5, 10 mg/kg/d
Basis:
actual ingested

No. of animals per sex per dose:

10 males and 20 females

Control animals:

yes, concurrent vehicle

Details on study design:

None stated

Positive control:

None stated

Parental animals: Observations and examinations:

None stated

Oestrous cyclicity (parental animals):

None stated

Sperm parameters (parental animals):

None stated

Litter observations:

None stated

Postmortem examinations (parental animals):

In each dose group half of the females were sacrificed on day 20 of pregnancy, the remaining pregnant animals were allowed to deliver. On sacrifice of pregnant rats, uterus, dead or viable fetuses and resorptions were observed; visceral and skeletal exam of fetuses was carried out.

Postmortem examinations (offspring):

After weaning, 20 females were randomly chosen and mated with non-consanguineous males (up to 10 weeks of age).
On day 20 of pregnancy, half of the females of this progeny, were necropsied and uterus and fetuses were examined for implantation sites, including viable, dead and/or resorbed fetuses.
On 10 females of each group dose was recorded the fertility and on day 20 of gestation the animals were sacrificed.

Statistics:

The results were statistically analyzed by Student's t-test for independent samples.

Reproductive indices:

None stated

Offspring viability indices:

None stated

Clinical signs:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

The test substance up to 10 mg/kg by s.c. route did not influence the fertility of parents.

Dose descriptor:

NOAEL

Effect level:

>= 10 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other: The test substance up to 10 mg/kg by s.c. route did not influence the fertility of parents.

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings:

not specified

The average numbers of viable fetuses, implantation sites, anomalies and weight of fetuses were in good agreement with the control group.

Reproductive effects observed:

not specified

Conclusions:

The studies on the fertility and on the reproductive performance in rats, carried out on F0, F1 and F2 progenies, showed that the test substance up to 10 mg/kg by s.c. route did not influence the fertility of parents and the pregnancy of dams; moreover in this study no effects on fetal and peri- and postnatal development of progeny were observed.

Effect on fertility: via dermal route

Endpoint conclusion:

no adverse effect observed

Study duration:

subacute

Species:

rat

Quality of whole database:

Limited study summary. Not GLP.

Additional information

Short description of key information:
Low molecular weight test substance given SC to rats at doses up to 10 mg/kg/d did not adversely affect fertility (male or female), conception, or fetal development (Bertoli, 1986).

Justification for selection of Effect on fertility via dermal route:
2 (reliable with restrictions)

Effects on developmental toxicity

Description of key information

The test substance administered to rats during organogenensis period, up to 10 mg/kg, showed no embryotoxic and fetotoxic activity (Bertoli, 1986). 

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Limited study summary. Not GLP.

Qualifier:

no guideline followed

Principles of method if other than guideline:

The Sprague-Dawley rats were used in this teratogenicity study. The test substance was administered into dorsal region by subcutaneous route once daily on days 5-16 of pregnancy.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Female 8 weeks of age, male 10 weeks of age
- Weight at study initiation: Female and male weighing 200±10 g
- Housing: The animals were housed in Makrolon cages in an air-conditioned room.
- Diet (e.g. ad libitum): Females received pelletted dry diet Altromin MT and males diet Altromin R.
- Water (e.g. ad libitum): Tap water was available ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-23°C
- Humidity (%): 55-60%
- Air changes (per hr): Ventilation was 12 times/h
- Photoperiod (hrs dark / hrs light): Lighting schedule was 12:12 light/dark (from 7:00 a.m. to 7:00 p.m.)

No additional data

Route of administration:

subcutaneous

Vehicle:

physiological saline

Details on exposure:

The test substance was dissolved in physiological solution and administered into dorsal region, by subcutaneous route, at 1-10 mg/kg at a volume of 5 mL/kg. For comparison was used conventional heparin (H) with activity of 163 USP/mg at 1-10 mg/kg by s.c. route (5 mL/kg).

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

- M/F ratio per cage: 1/3
- Length of cohabitation: one night
- Proof of pregnancy: sperm in vaginal smear referred to as day 1 of pregnancy

No additional data

Duration of treatment / exposure:

On days 5-16 of pregnancy.

Frequency of treatment:

Once daily

Duration of test:

21 days

Remarks:

Doses / Concentrations:
1, 10 mg/kg
Basis:
actual ingested

No. of animals per sex per dose:

10 females

Control animals:

yes, concurrent vehicle

Details on study design:

None stated

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: During the gestation the animals were daily observed for any changes in appearance or behaviour and mortality.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: During the gestation the animals were daily observed for any changes in appearance or behaviour and mortality.

BODY WEIGHT: Yes
- Time schedule for examinations: Body weight was measured at selected intervals (1, 7, 14, 21 days).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21
- Organs examined: The uterus and the fetuses were examined, the number and location of viable, dead and resorbed fetuses were recorded.

No additional data

Ovaries and uterine content:

None stated

Fetal examinations:

Viable fetuses were weighed, examined externally and the cranial, thoracic and abdominal organs were examined by dissection.
The skeletons were examined after they had been fixed in 70% alcohol, cleared with potassium hydroxide and stained with alizarin red-S.

Statistics:

The results were statistically analyzed by Student's t-test for independent samples.

Indices:

None stated

Historical control data:

None stated

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
The test substance administered to rats during organogenensis period, up to 10 mg/kg, showed no embryotoxic and fetotoxic activity, since the drugs did not make a change, in comparison to the controls, in number of implantation sites, resorptions and fetal mortality.

Dose descriptor:

NOAEL

Effect level:

>= 10 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: other:

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Visceral and skeletal macroscopic alterations did not appear in the living fetuses.
No differences from controls were reported in appearance, behaviour or body weight of the dams up to 10 mg/kg/d.

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

The test substance administered to rats during organogenensis period, up to 10 mg/kg, showed no embryotoxic and fetotoxic activity, since the drugs did not make a change, in comparison to the controls, in number of implantation sites, resorptions and fetal mortality.

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no adverse effect observed

Study duration:

subchronic

Species:

rat

Quality of whole database:

Limited study summary. Not GLP.

Additional information

Justification for selection of Effect on developmental toxicity: via dermal route:
2 (reliable with restrictions)

Justification for classification or non-classification

Based on the outcome of the studies the test substance does not need to be classified for reproductive and/or developmental effects.