A mixture of: sodium/potassium 7-[[[3-[[4-((2-hydroxy-naphthyl)azo)phenyl]azo]phenyl]sulfonyl]amino]-naphthalene-1,3-disulfonate

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 November 2013 to 28 October 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Specific details on test material used for the study:

Name: FAT 45155/E TE
Batch No.: BS-DUW 1120/004-01
Physical State: solid (powder)
Storage Conditions: room temperature
Expiry Date: 15 August 2018

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test System
Species/strain: healthy Wistar rats
Source: Charles River, 97633 Sulzfeld, Germany
Sex: male and female; the female animals were non-pregnant and nulliparous.
Age at the start of the treatment period: males: 11-12 weeks old, females: 11-12 weeks old.
Body weight at the allocation of the animals to the experimental groups: males: 303 - 345 g (mean: 327.48 g, ± 20% = 261.98 – 392.97 g); females: 192 - 220 g (mean: 207.70 g, ± 20 % = 166.16 – 249.24 g)
The animals were derived from a controlled full-barrier maintained breeding system (SPF). According to Art. 9.2, No. 7 of the German Act on Animal Welfare the animals were bred for experimental purposes.
The animal study was authorized by local government under file no. 55.2-1-54-2532.2-11-11.

Housing and Feeding Conditions
- Full barrier in an air-conditioned room
- Temperature: 22 ± 3 °C
- Relative humidity: 55 ± 10 %
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 0801)
- Free access to tap water, sulphur acidified to a pH of approximately 2.8 (drinking water, municipal residue control, microbiological controls
at regular intervals)
- The animals were kept individually in IVC cages (except during the mating period when one female will be paired with one male), type III H,
polysulphone cages on Altromin saw fibre bedding (lot no. 060613)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least 5 days) under laboratory conditions

Preparation of the Animals
Prior to the start of the treatment period a detailed clinical observation outside the home cage was made. 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.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

Preparation of the Test Item and Control Formulations
The test item was weighed into a tarred plastic vial on a precision balance and the vehicle was added to give the appropriate final concentration of the test item. The test item was dissolved in aqua ad injectionem. The vehicle was selected as suggested by the sponsor based on the test item’s characteristics and testing guideline. The test item and control formulations were prepared freshly on each administration day before the administration procedure. Homogeneity of the test item in the vehicle was maintained by vortexing the prepared suspension thoroughly before every dose administration. The vehicle was also used as control item.

Experimental Groups and Doses
The animals were treated with the test item formulation or vehicle on 7 days per week for a period of 54 days, i.e. during 14 days of pre-mating and maximum 14 days of mating in both males and females, during the gestation period and up to post-natal day 3 in females. Males were dosed after the mating period until the minimum total dosing period of 28 days was completed. In consultation with the sponsor the following doses were selected for the 3 dose groups (LD = low dose, MD = medium dose, HD = high dose) and 1 control group (C).


C 0 mg/kg bw/ day (Male No.:1-10/ Female No.:41-50)
LD 100 mg/kg bw/ day (Male No.:11-20/ Female No.:51-60)
MD 300 mg/kg bw/ day (Male No.:21-30/ Female No.:61-70)
HD 1000 mg/kg bw/ day (Male No.:31-40/ Female No.:71-80)

The highest dose level was chosen with the aim of inducing toxic effects, but no death or severe suffering. Thereafter, a descending sequence of dose levels was selected with a view to demonstrate any dosage related response and NOAEL. The animals in the control group were handled in an identical manner to the test group subjects and received aqua ad injectionem using the same volume as used for the high dose group.

Administration of Doses
The test item and vehicle were administered at a single dose to the animals by oral gavage. The application volume for all groups was 10 mL/kg body weight. For each animal the individual dosing volume was calculated on the basis of the body weight most recently measured.

Details on mating procedure:

Mating was performed using a ratio of 1:1 (male to female). The vaginal smear of the females was checked every morning after the start of the mating period to confirm the pregnancy. If the vaginal smear of a particular female was not found to be sperm-positive, the actual stage of the estrus cycle on that day was documented. The day of the vaginal plug and/or sperm was considered as day 0 of gestation. The cages were arranged in such a way that possible effects due to cage placement were minimised.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Each dosing concentration was analysed with respect to the target nominal concentration. Stability and homogeneity of the test item in the vehicle were analysed for the low and high dose concentrations. Samples for the nominal concentration verification were taken in study week 1 (first week of pre-mating period), 3 (first week of mating), 5 (gestation) and in the last week of the study (gestation/lactation) from all groups (16 samples). Samples for homogeneity analysis were taken from the top, middle and bottom of the high dose and the low dose formulation in study week 1 and 5 (12 samples). Samples for stability analysis were taken in the first week of the study, 0 hours after the preparation and another sample 6 hours after the preparation (at room temperature), from high and low dose formulations (4 samples). Each sample was retained twice (sample A, sample B, each of at least 25 mL). All formulation samples were stored at -20 °C. The A samples were analysed after the completion of the toxicity study at BSL BIOSERVICE Scientific Laboratories GmbH under the BSL study no. 135627. The B samples will be discarded after completion of the final study report. The exact procedure was described in a phase plan which was amended to the study plan. The results are reported in the appendix of the final report.

Duration of treatment / exposure:

The animals were treated with the test item formulation or vehicle on 7 days per week for a period of 54 days, i.e. during 14 days of pre-mating and maximum 14 days of mating in both males and females, during the gestation period and up to post-natal day 3 in females. Males were dosed after the mating period until the minimum total dosing period of 28 days was completed.

Frequency of treatment:

7 days/week

Details on study schedule:

Arrival of the Test Item: 23 September 2013
Study Initiation Date: 04 November 2013
1st Amendment to Study Plan: 14 November 2013
2nd Amendment to Study Plan: 20 January 2014
3rd Amendment to Study Plan: 29 January 2014
4th Amendment to Study Plan: 25 March 2014
Experimental Starting Date: 15 November 2013
Experimental Completion Date: 07 January 2014
Completion Date of Delegated Phase (Histopathology): 06 October 2014
Completion Date of Delegated Phase (Formulation Analysis): 17 March 2014

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day

Remarks:

Control

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Low dose

Doses / concentrations 3

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Middle dose

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

High dose

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

Animals were allowed to mate in 1:1 male to female ratio. The vaginal smear of the females was checked every morning after the start of the mating period to confirm the pregnancy. If the vaginal smear of a particular female was not found to be sperm-positive, the actual stage of the estrus cycle on that day was documented. The day of the vaginal plug and/or sperm was considered as day 0 of gestation. Females with unsuccessful mating were allowed to mate with other male of the same group. The cages were arranged in such a way that possible effects due to cage placement were minimised.

Examinations

Parental animals: Observations and examinations:

Clinical Observations
General clinical observations were made at least once a day, preferably at the same time each day and considering the peak period of anticipated effects after dosing. The health condition of the animals was recorded. Twice daily all animals were observed for morbidity and mortality except on weekends and public holidays when observations were made once daily. Females showing signs of abortion or premature delivery prior to the scheduled scarification of the animals were sacrificed and subjected to a thorough macroscopic examination. Clinical observations included spontaneous activity, lethargy, recumbent position, convulsions, tremors, apnoea, asphyxia, vocalisation, diarrhoea, changes in the skin and fur, eyes and mucous membranes (salivation, discharge), piloerection and pupil size. Changes in gait, posture, response to handling as well as the presence of clonic or tonic movements, stereotypes, difficult or prolonged parturition or bizarre behaviour were recorded.

Body Weight and Food Consumption
The body weight was recorded once before the assignment to the experimental groups, on the first day of administration and weekly during the treatment period as well as at the end of the study. During pregnancy, females were weighed on gestation days (GD) 0, 7, 14 and 20 and within 24 hours of parturition (day 0 post-partum) as well as day 4 post-partum along with pups. Food consumption was measured weekly on the corresponding days of the body weight measurements after the beginning of the dose administration. Food consumption was not measured during the mating period in males and females and the post-mating period in males.

Litter observations:

The duration of the gestation was recorded and was calculated from day 0 of the pregnancy. Each litter was examined as soon as possible after delivery of the dam to establish the number and sex of pups, stillbirths, live births, runts and the presence of gross abnormalities. Live pups were counted and sexed and litters weighed within 24 hours of parturition (day 0 post-partum) and on day 4 post-partum. Live pups were identified by writing numbers on the back with the help of a permanent marker or by tattooing. In addition to the observations of parent animals, any abnormal behaviour of the offspring was recorded.

Postmortem examinations (parental animals):

Pathology
All male animals were sacrificed after the completion of the mating period (total dosing period: 28 days) on study day 29, while female animals were sacrificed on post-natal day 4 along with pups using terminal anaesthesia (ketamine/xylazin, 2:1, Pharmanovo, lot no: 24664, expiry date: 06/2015 and Serumwerk, lot no: 00512, expiry date: 07/2014).
Pups sacrificed on day 4 post-partum were carefully examined externally for gross abnormalities.
Non-pregnant females were sacrificed on study day 26 using the sperm-positive vaginal smear as an evidence of mating. All animals were subjected to a detailed gross necropsy which includes careful examination of the external surface of the body, all orifices and the cranial, thoracic and abdominal cavities and their contents. Special attention was paid to the organs of the reproductive system. The ovaries, uterus with cervix, vagina, testes, epididymites, accessory sex organs (prostate, seminal vesicles with coagulating glands as a whole), and all organs showing macroscopic lesions of all adult animals were preserved in 10 % neutral buffered formalin, except for testes and epididymites which were fixed in Modified Davidson’s Solution for approximately 24 hours before they were transferred to 70 % ethanol. The number of implantation sites and corpora lutea was recorded for each parental female at necropsy. The number of corpora lutea and implantation sites was recorded for any females sacrificed 24 to 26 days after the end of the pairing period with no evidence of mating and for any females sacrificed on day 25 post-coitum due to non-delivery.

Organ Weights
The testes and epididymites of all male adult animals as well as the ovaries, uterus with cervix of all female adult animals were weighed. Paired organs were weighed together.

Histopathology
Testes, epididymites, ovaries, uterus with cervix, vagina, accessory sex organs (prostate, seminal vesicle with coagulating gland) were examined in Control and HD animals and in non-pregnant female animal no. 69 of the MD group. Testes, epididymites and accessory sex organs (prostate, seminal vesicle with coagulating gland) were also examined in the mating partners of the non-pregnant female no. 69 of the MD group. Any gross lesion macroscopically identified was examined microscopically in all animals. Discoloration possibly due to the test item was evaluated in the organs of all dose groups. For the testes, a detailed qualitative examination was made; taking into account the tubular stages of the spermatogenic cycle at evaluation of additional hematoxylin-PAS (Periodic Acid Schiff) stained slides. Histological processing of tissues to microscope slides was performed at the GLP-certified contract laboratory AnaPath GmbH, AnaPath Services, Hammerstrasse 49, 4410 Liestal, Switzerland (test site for tissue processing). Histopathological evaluation was performed at the GLP-certified contract laboratory AnaPath GmbH, Buchsweg 56, 4625 Oberbuchsiten, Switzerland (test site for histopathology). The study phases from test site 1 and 2 was performed in compliance with the Swiss Ordinance relating to Good Laboratory Practice adopted 18 May 2005 [SR 813.112.1] (Status as of 01 December 2012). Blocking, embedding, cutting, H&E staining and scientific slide evaluation were performed according to the corresponding SOP’s of the test sites. The principal investigator for histopathological tissue processing sent all raw data (including blocks, slides, paper raw data, statement of compliance and quality assurance statement) to the study director. The principal investigator for histopathological evaluation provided the histopathology results to the study director by e-mail and sent a pathology phase report to the study director upon the completion of the study.

Postmortem examinations (offspring):

All surviving pups were sacrificed by decapitation on post-natal day 4. Dead pups and pups sacrificed on day 4 post-partum were carefully examined externally for gross abnormalities.

Statistics:

A statistical assessment of the results of the body weight, food consumption, parameters of absolute and relative organ weights were performed for each gender by comparing values of dosed with control animals of the main groups using a one-way ANOVA and a post-hoc Dunnett Test.
These statistics were performed with GraphPad Prism V.6.01 software (p<0.05 was considered as statistically significant).

Reproductive indices:

There were no statistically significant effects on reproductive indices in this study.
A slightly reduced fertility index (number of pregnant females / number of copulated females X 100) was observed in the LD group (90%, respectively) as compared to 100% in the control group. This is considered incidental, as it is within the normal range of variation for this strain.
The cause of the unsuccessful pregnancy and/or infertility of female no. 51 could not be established from the reproductive organs examined
histopathologically from this animal.

Offspring viability indices:

The viability index (No. of live offspring at day 4 / No. of live offspring at birth) was slightly reduced in the HD group (95 %) when compared with the control group (100 %). As values were within the normal range of variation of historical control data, this is considered incidental.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Moving the bedding was noted on one or two days in few male animals of all dose groups with dose-dependently increasing incidences. In female animals moving the bedding was observed mainly in the LD group but also in 3/10 animals of HD group. Occasional incidences of slight salivation were observed in 1/10 female of LD and 2/10 females of HD group, but not in male animals. As the symptoms of salivation and moving the bedding were noted mainly immediately after administration of test item, these signs are considered to be a local effects caused by the bolus administration of the test item formulation and are not considered to be signs of systemic toxicity. Regurgitation of the test item formulation was noted on single or two consecutive days in single animals of the LD and MD group. This was not associated with any other findings but moving the bedding and is not assumed to be related to a systemic effect of the test item.
Mild clinical symptoms, like nasal discharge, eschar and alopecia at various parts of the body were observed in few isolated male and female animals of control and dose groups. These findings were mostly transitory in nature and are considered to be incidental. Red urine (haematuria) was observed in male animal no. 31 of the HD group on a single pre-mating day. This observation was transitory and is not considered to be of toxicological relevance.
Moderate piloerection was noted in male animal no. 36 of the HD group on a single mating day and is assumed to be an incidental finding.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

In both males and females, the mean body weight increased with the progress of the study in the control, the LD, the MD and the HD group. Values were within the normal range of variation of historical control data. There were no statistically or biologically significant effects in body weight and body weight gain of both males and females of the dose groups, when compared to the respective controls.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No considerable effect of FAT 45155/E on food consumption was found in any of the groups of male and female animals and statistically significant differences are not assumed to be biologically relevant. A slight – but statistically significant - tendency towards higher food consumption was observed in the male animals of the MD group in the second week of premating compared to the control group. This slight difference is not assumed to be toxicologically relevant as values were within the normal range. In the first week of gestation, a slightly – but not statistically significantly – higher food consumption was observed for the females of HD group
compared to the control group. This slight difference is not assumed to be toxicologically relevant.

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

There were not treatment-related effects on the completeness of stages or cell populations of the testes. Some histologic alterations were recorded at a minimum and/or slight severity in a few animals from both of the control and high-dose males and in male no. 29 from group 3, but all of these were considered to be within the range of normal background lesions which are commonly observed in males of this strain and age. Nothing special was observed in ovaries, uterus with cervix and vagina from female no. 69 which failed to produce a pregnancy. In the mating partner male no. 29, minimal mononuclear cell foci in epididymites as well as minimal spermatid retention in testes was observed, but these were the findings which are commonly observed in males of this strain and age, and therefore, were considered not to be related to the non-pregnancy of female no. 69. At the necropsy, “Distended, hard, discoloured” uterus was recorded in female no. 51 (group 2), which correlated microscopically with inflammation with purulent contents in the lumen. Such inflammatory change, except for the normal reaction at/around the implantation site, was not observed in any high-dose females and in female no. 69 of group 3. Therefore, the lesion found in female no. 51 was an incidental change and considered not to be related to treatment with the test item. The remainder of findings recorded was within the range of normal background lesions which may be recorded in animals of this strain and age.

Description (incidence and severity):

Dose Formulation Analysis:
Concentration analysis of formulation samples was determined in study week 1, 3, 5 and 7 for all dose groups. The mean recoveries observed in LD, MD and HD groups were 89.4 %, 96.5 % and 91.1 % of the nominal concentration, respectively. Stability of formulation samples was investigated for concentration of HD and LD dose groups. For HD formulation six hour stability was missed and hence not confirmed. After 6 hours storage at room temperature recovery compared to starting value was between 75.2 % and 108.4 %. On the basis of the recovery at 0h and 6h it can be assumed that in HD group 68.5 to 91.1 % were applied within the specified duration of 6 hours after preparation. Homogeneity of formulation samples was determined in study week 1 and 5 for LD and HD dose groups. The mean recovery observed for LD dose group was between 77.5 % and 91.8 % of the nominal value and between 78.8 and 97.4 % of the nominal value for HD dose group. The coefficients of variation of the different sampling locations (top, middle, bottom) were between 0.5 % and 15.5 % in LD dose group and between 5.5 % and 9.3 % in HD dose group.

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

Reproductive Indices
There were no statistically significant effects on reproductive indices in this study. A slightly reduced fertility index (number of pregnant females / number of copulated females X 100) was observed in the LD group (90 %, respectively) as compared to 100 % in the control group. This is considered incidental, as it is within the normal range of variation for this strain. The cause of the unsuccessful pregnancy and/or infertility of female no. 51 could not be established from the reproductive organs examined histopathologically from this animal. The viability index (No. of live offspring at day 4 / No. of live offspring at birth) was slightly reduced in the HD group (95 %) when compared with the control group (100 %). As values were within the normal range of variation of historical control data, this is considered incidental.

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

ca. 685 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Study findings

Remarks on result:

other: On the basis of the recovery at 0h and 6h it can be assumed that in HD group 68.5 to 91.1 % were applied within the specified duration of 6 hours after preparation.

Results: F1 generation

General toxicity (F1)

Mortality / viability:

no mortality observed

Sexual maturation:

not examined

Effect levels (F1)

Remarks on result:

not measured/tested

Overall reproductive toxicity

Reproductive effects observed:

no

Applicant's summary and conclusion

Conclusions:

The NOAEL for general and reproduction toxicity in this study is considered to be at lowest 685 mg/kg/d.

Executive summary:

In a GLP-compliant study was carried out to assess the possible effects of FAT 45155/E on male and female fertility and embryofetal development after repeated dose administration in Wistar rats according to OECD guideline 421. The test item was administered daily in graduated doses to 3 groups of test animals, one dose level per group for a treatment period of 54 days, i.e. during 14 days of pre-mating and maximum 14 days of mating in both males and females, during the gestation period and up to post-natal day 3 in females. Males were dosed after the mating period until the minimum total dosing period of 28 days were completed. Animals of an additional control group were handled identically as the dose groups but receivedaqua ad injectionem, the vehicle used in this study. The 4 groups comprised 10 male and 10 female Wistar rats.

The following doses were evaluated:

Control:                        0        mg/kg/d

Low Dose:                    100     mg/kg/d

Medium Dose:              300     mg/kg/d

High Dose:                   1000   mg/kg/d

 

The test item formulation was prepared freshly on each day of administration. The test item was dissolved in aqua ad injectionem and administered daily during 14 days of pre-mating and 14 days of mating in both male and female animals, during the gestation period and up to post-natal day 3 in females. Males were dosed for 28 days. Dose volumes were adjusted individually based on weekly body weight measurements. The administration volume was 10 mL/kg body weight. During the period of administration, the animals were observed each day for signs of toxicity. Animals that died were examined macroscopically and at the conclusion of the test, surviving animals were sacrificed and observed macroscopically. Body weight and food consumption were measured weekly, except for food consumption measurements which were not taken during the mating period in female animals and the mating and post-mating period in male animals. After 14 days of treatment to both male and female, animals were mated (1:1) for a maximum of 14 days. The subsequent morning onwards the vaginal smears of females were checked to confirm the evidence of mating. After the confirmation of the mating, females were separated and housed individually. Each litter was examined as soon as possible after delivery of the dam to establish the number and sex of pups, stillbirths, live births, runts and the presence of gross abnormalities. Live pups were counted, sexed and litters weighed within 24 hours of parturition and on day 4 post-partum. The males were sacrificed after completion of the mating period on treatment day 29 and the females along with their pups were sacrificed on post natal day 4. Non-pregnant females were sacrificed on day 26. The number of implantation sites and corpora lutea was recorded for each parental female at necropsy. Pups sacrificed on postnatal day 4, were carefully examined for gross external abnormalities. A full histopathological evaluation of testes, epididymides, ovaries, uterus with cervix, vagina and accessory sex organs (prostate, seminal vesicle with coagulating gland) was performed on high dose and control animals and in non-pregnant female animal no. 69 of the MD group and their male mating partners. Any gross lesion macroscopically identified was examined microscopically in all animals. No mortality occurred in the control or in any of the dose groups during the treatment period of this study. Clinical findings suggest a slight transient local effect of FAT 45155/E immediately after administration via oral gavage.Clinical signs of systemic toxicity were not observed. There was no test item related effect on body weight and body weight change in both, males and female animals. No effect of test item on food consumption of male and female animals was found at any of the dose levels tested. There were no test item-related effects on litter data recorded in this study. Parameters, like total number of pups born, number of live pups, still births and runts on PND 0 as well as number of male pups, number of female pups and sex ratio on PND 0 and PND 4 were not significantly different between dose groups and control group. There were no FAT 45155/E-related effects on the litter weight data, including pup mean weight, total litter weight, male and female litter weight on PND 0 and PND 4. No test item-related effect was observed on the pre-coital interval or in the duration of gestation when compared with the control group. All pregnancies resulted in normal births. There were no test item related effects on the number of corpora lutea, number of implantation sites, number of live pups (PND 0 and PND 4) and percentage of pre- and post-implantation loss in this study. There were no test item related effects on reproductive indices in this study. There was no test item related effect on the survival of the pups from PND 0 through PND 4 in this study. No test item related gross external abnormalities were observed in males, females or pups in any of the dose groups of this study. Slightly but statistically significantly lower ovary weight at 1000 mg/kg/d was not associated with histomorphological abnormalities and is not assumed to be toxicologically relevant. FAT 45155/E produced no histological evidence of toxicological properties in the organs and tissues of the reproductive system; i.e. testes, epididymides, prostate, seminal vesicles, coagulating glands, ovaries, uterus and cervix, and vagina. In addition, as a result of the detailed examination of the testis, it was judged that there were no treatment-related effects on the testicular histomorphology including spermatogenesis and interstitial cell structure. Correct concentrations of FAT 45155/E in dose formulations prepared in study week 1, 3, 5 and 7 were confirmed, as values deviated less than 20 % (10.6 %, 3.5 % and 8.9 % for LD, MD and HD, respectively) from nominal value. Dose formulations of the LD and HD groups were considered to be homogeneous as the COV was below the acceptance criteria of 20 % (0.5 and 15.5 % for LD group and 5.5 and 9.3 % for HD group). Six hour benchtop stability was missed and hence not confirmed for the HD formulation. Whereas in LD formulation a recovery of 108.4 % was found at 6 h, compared to 0 h, in HD formulation the recovery at 6 h was only 75.2 %. This difference in HD formulation was below acceptance criteria of 20 %. On the basis of this reproduction/developmental toxicity screening test with FAT 45155/E in male and female Wistar rats with dose levels of 100, 300, and 1000 mg/kg/d the no effects of FAT 45155/E were found at the dose levels used in this study. The NOEL of FAT 45155/E in this study is considered to be the high dose used. Considering the results on dose confirmation observed in this study, the NOAEL for general and reproduction toxicity in this study is considered to be at lowest 685 mg/kg/d.