Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
fertility, other
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2014-03-27 to 2015-06-15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
other: OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents), adopted 21 September 1998
Deviations:
no
Qualifier:
according to
Guideline:
other: EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Guanidinhydrochloride

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Wistar rats, Crl: WI(Han) (Full Barrier)
- Source: Charles River, 97633 Sulzfeld, Germany
- Age at study initiation: females: 7-8 weeks old, males: 7-8 weeks old
- Weight at study initiation: females: 137 -170 g; males: 147 -193 g
- Housing: The animals were housed in groups (5 animals/sex/cage) in type IV cages
- Diet (e.g. ad libitum): Altromin 1324 maintenance diet for rats and mice (lot no. 1526) ad libitum
- Water (e.g. ad libitum): Free access to tap water, sulphur acidified to a pH of approximately 2.8
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 55 +/- 10 %
- Air changes (per hr): 10 x / hour
- Photoperiod (hrs dark / hrs light): Artificial light, sequence being 12 hours light, 12 hours dark

IN-LIFE DATES: males From: 2014-03-25 to 2014-08-05 females From: 2014-03-25 to 2014-08-04

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
VEHICLE
-aqua ad injectionem (AlleMan Pharma)
- Concentration in vehicle: low dose: 5 mg/L; medium dose 20 mg/mL; high dose: 60 mg/mL
- Amount of vehicle (if gavage): dose volume for all groups was 5 mL/kg body weight
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
For determination of the concentration of test item in dosing formulations, samples of at least 5 mL were retained from all groups in weeks 1, 5, 9 and 13
during the treatment period and stored between -15 and -35 °C. In total 16 samples.
Stability of the dosing formulations was tested once at the beginning of the treatment period.
From the low, medium and high dose group, samples of dosing formulations were frozen after 0 hours and after 10 days (at room temperature) after the preparation and stored at -15 to -35 °C until analysis. In total 6 samples.
Duration of treatment / exposure:
90 days
Frequency of treatment:
7 days per week
Details on study schedule:
Fertility parameters from a subchronic repeated dose toxicity study
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
25 mg/kg bw
Basis:
actual ingested
Remarks:
Doses / Concentrations:
100 mg/kg bw
Basis:
actual ingested
Remarks:
Doses / Concentrations:
300 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
control: 15 animals per sex
high dose: 15 animals per sex
low dose: 10 animals per sex
mid dose: 10 animals per sex
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: according to the results of a previous dose range finding study
- Rationale for animal assignment (if not random): Before the first administration all animals used for the study were weighed and assigned to the experimental groups with achieving a most homogenous variation in body weight throughout the groups of males and females, respectively, while ensuring to keep each animal with its initial cage partners.
- Rationale for selecting satellite groups: In order to allow a detection of possible delayed occurrence or persistence of or recovery from toxic effects, the animals in the recovery groups were observed for a period of 28 days (females) or 29 days (males) following the last administration.
Positive control:
no

Examinations

Parental animals: Observations and examinations:
Refer to entry under IUCLID section 7.5.1
Oestrous cyclicity (parental animals):
Daily over a period of 8 days, the estrous cycle of all female animals was examined 4, 8 and 12 weeks after the first administration.
In the recovery animals the estrous cycle was examined during the last week of the recovery period.
Sperm parameters (parental animals):
At necropsy (one day after the last administration) and at the end of the recovery period, left epididymis, left testis and left vas deferens were separated and used for evaluation of sperm parameters.
Epididymal sperm motility and testicular sperm count were evaluated in all male animals using Hamilton Thorne Sperm Analyser
(TOX IVOS Version 13.0).
Therefore sperm from left vas deferens was transferred to 0.1% bovine serum albumin solution. For staining two drops of 1% aqueous Eosin-Y solution were mixed with six drops of the sperm-suspension. The stained sperm suspension was used to prepare smears on slides. After complete drying the slides were dipped into 0.1% acetic acid for approximately 30 seconds to intensify the colouring.

Results and discussion

Results: P0 (first parental animals)

Details on results (P0)

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
Guanidinhydrochloride had no biologically significant effect on the estrous cycle analyzed 4, 8 and 12 weeks after the first administration and in the last week of the recovery period. There were no considerable differences in the length or sequence of cycle stages between the dose groups and the control group. Deviations from the physiological 4 or 5 day cycle in the rat were observed occasionally, mainly as irregularly long cycles, in all treatment groups including control and irrespective of the duration of the treatment. This was considered incidental and not related to the treatment with the test item.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
Guanidinhydrochloride had no effect on epididymal sperm motility or testicular sperm count analyzed at the end of the treatment or recovery period of this study. The statistical analysis showed no statistically significant changes between the control group and any of the dose groups neither in the percentage of motile, static or rapidly moving epididymal sperms nor in testicular number of sperms/g testis.
Sperm staging and evaluation of sperm morphology did not reveal any indicator for toxicity induced by the test item.

For other parameters refer to IUCLID section 7.5.1 Robust Study Summary for repeated dose toxicity

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Executive summary:

In a subchronic toxicity study according to OEDC Guideline 408 (adopted 21 September 1998), Guanidine hydrochloride was administered to 10 Wistar rats/sex/dose in water, by gavage at dose levels of 0, 50, 100 and 300 mg/kg bw/day.

In order to allow a detection of possible delayed occurrence or persistence of or recovery from toxic effects a satellite group of 5 rats/sex was exposed at dose levels of 0 and 300 mg/kg bw/day (control and HD).

To evaluate possible toxic effects on fertility, the estrous cycle was examined at defined time points of the treatment and recovery period and epididymal sperm motility, testicular sperm count and sperm morphology from vas deferens were evaluated at the end of the treatment and recovery period. Moreover, a detailed histopathological evaluation of the reproductive organs was performed.

Guanidinhydrochloride had no effect on epididymal sperm motility or testicular sperm count analyzed at the end of the treatment or recovery period of this study. Sperm staging and evaluation of sperm morphology did not reveal any indicator for toxicity induced by the test item. Guanidinhydrochloride had no biologically significant effect on the estrous cycle analyzed 4, 8 and 12 weeks after the first administration and in the last week of the recovery period.