Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information
Not applicable
Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 12 August 2013 to 27 January 2014.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducetd according to international test guidelines and to GLP.
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
: the estrous cycle stage was not determined on the third day of the mating period for one control female, due to an oversight. This deviation was considered not to have compromised the validity or integrity of the study.
Principles of method if other than guideline:
Not applicable.

GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France.
- Age at study initiation: males were approximately 10 weeks old and the females were approximately 9 weeks old.
- Weight at study initiation: males had a mean body weight of 408 g (range: 367 g to 449 g) and the females had a mean body weight of of 225 g (range: 208 g to 252 g).
- Fasting period before study: no
- Housing: the animals were individually housed, except during pairing and lactation, in polycarbonate cages (Tecniplast 2154, 940 cm²) with stainless steel lids and containing autoclaved sawdust (SICSA, Alfortville, France). Individual housing was chosen since it is preferable for pregnant animals and mating purpose.
Toward the end of gestation and during lactation, autoclaved wood shavings (SICSA, Alfortville, France) were provided to females and their litter as nesting material.

- Diet (e.g. ad libitum): SSNIFF R/M-H pelleted maintenance diet, batch Nos. 3833086 and 1229567 (SSNIFF Spezialdiäten GmbH, Soest, Germany)
- Water (e.g. ad libitum): tap water (filtered with a 0.22 µm filter).
- Acclimation period: 7 days before the beginning of the treatment period. A larger number of animals than necessary were acclimated to permit the selection and/or replacement of individuals.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): about 12 cycles/hour of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12h/12h.

IN-LIFE DATES: From: 3 September 2013 to 03 November 2013.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test item dose formulations were prepared daily and delivered to the study room at room temperature and protected from light. No correction factor was applied for the dose calculations.

VEHICLE
- Amount of vehicle (if gavage): 5 mL/kg bw/day
- Lot/batch no. (if required): MKBH4894V and MKBP7039V.
- Purity: no data
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: up to 15 days
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged individually.
- Any other deviations from standard protocol: none

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Concentrations of the test item in the dose formulations have been quantified by a validated Ion Chromatography (IC) with conductivity detection analytical method.
The validation of the analytical method was conducted in CiToxLAB France/Study No. 40285 VAA and precise details concerning the checked parameters, acceptance criteria and obtained results are documented in the corresponding validation report.
Duration of treatment / exposure:
In the males:
- 2 weeks before mating (from study day 1 to 14)
- during the mating period (from study day 15 until study day 16 to 19),
- until sacrifice (at least 5 weeks in total) (from study day 17 to 20 until study day 36).

In the females:
- 2 weeks before mating (from study day 1 to 14),
- during the mating period (from study day 15 until study day 16 to 19),
- during gestation (from study day 16 to 19 until study day 36 to 40),
- during lactation until day 5 p.p. inclusive (from study day 37 to 41 until study day 42 to 46),
- until sacrifice for the non-pregnant female (until study day 42).

Day 1 corresponds to the first day of the treatment period.
Frequency of treatment:
Once daily, at approximately the same time (7 days/week)

Details on study schedule:
- Age at mating of the mated animals in the study: 12 weeks for males and 11 weeks for females
Remarks:
Doses / Concentrations:
0, 30, 100 or 300 mg/kg bw/day.
Basis:
nominal conc.
No. of animals per sex per dose:
10 animals/sex/dose.

Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: the dose-levels were set based on the results of a previous 2-week dose-range finding study performed in the same species and strain (CiToxLAB France/Study No. 40287 TSR). ). In this study, 4 groups of 3 males and 3 females received 0 (corn oil), 100, 400 or 1000 mg/kg/day daily for 2 weeks by gavage with a constant dose volume of 5 mL/kg/day. In this preliminary study, there were the following findings:
- clinical signs: ptyalism (hypersalivation) from 400 mg/kg/day (from study day 3 or 8) in both sexes,
- body weight and food consumption: no obvious effects (high variations between animals),
- macroscopic examination at necropsy: no effects on mean organ weight, but thickened and/or white discoloration of forestomach from 400 mg/kg (1 to 3 animals per sex).
Ptyalism was considered to be test item treatment-related but of minor toxicological significance. However, the effects on forestomach were considered to represent signs of local intolerance to the test item and potentially related to its corrosive properties. Therefore, 300 mg/kg/day was selected as the high-dose level. The low-dose and mid-dose were selected using a ratio representing approximately a 3-fold interval (i.e. 30 and 100 mg/kg/day).

- Rationale for animal assignment (if not random): during the acclimation period, the required number of animals (40 males and 40 females) was selected according to body weight and clinical condition. The animals were allocated to groups (by sex) using a computerized stratification procedure based on body weight, so that the average body weight of each group was similar.

Positive control:
Not used.

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: each animal was checked for mortality or signs of morbidity once a day before the treatment period and at least twice a day during the treatment period, including weekends and public holidays. From arrival, each animal was observed once a day as part of routine examinations. From the start of the treatment period, each animal was observed once a day, at approximately the same time, for the recording of clinical signs.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: detailed clinical examinations were performed on all animals once before the beginning of the treatment period and then once a week until the end of the study.
Observations included (but were not limited to) changes in the skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lachrymation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypes (e.g. excessive grooming, repetitive circling) or bizarre behavior (e.g. self mutilation, walking backwards) were also recorded.

BODY WEIGHT: Yes
- Time schedule for examinations: the body weight of each male was recorded on the first day of treatment (day 1), then once a week until sacrifice.
The body weight of each female was recorded on the first day of treatment (day 1), then once a week until mated and on days 0, 7, 14 and 20 post-coitum (p.c.) and days 1 and 5 p.p..

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
The quantity of food consumed by each male was measured once a week, over a 7 day period, from the first day of treatment until the start of the mating period. The quantity of food consumed by each female was measured once a week, over a 7 day period, from the first day of treatment until the start of the mating period, during pregnancy at the intervals days 0-7, 7 14 and 14-20 p.c. and during lactation for interval days 1 5 p.p.. During the mating period, food consumption was not measured for males or females.
Food intake per animal and per day was calculated by noting the difference between the food given and that in the food-hopper the next time.

Other: See section 7.5.1 (Repeated dose toxicity (Rep. dose tox. oral V1 CiToxLAB 2013)


Oestrous cyclicity (parental animals):
The estrous cycle stage was determined from a fresh vaginal lavage, each morning during the mating period, until the females were mated excepted on the third day for one control female.

Sperm parameters (parental animals):
Parameters examined in P1 male parental generation: testis weight and epididymis weight.

Litter observations:
STANDARDISATION OF LITTERS
Not applicable.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, live births, postnatal mortality, presence of gross anomalies, weight gain and physical or behavioural abnormalities.

GROSS EXAMINATION OF DEAD PUPS:
A macroscopic post-mortem examination of the principal thoracic and abdominal organs was performed on all found dead pups. Special attention was paid to the reproductive organs and to whether the pup had fed (e.g. presence of milk in the stomach).
Postmortem examinations (parental animals):
SACRIFICE
On completion of the treatment period, after at least 14 hours fasting, all F0 animals were deeply anesthetized by an intraperitoneal injection of sodium pentobarbital and sacrificed by exsanguination:
- Male animals: after the end of the pairing period (at least 5 weeks of treatment in total).
- Maternal animals: on Day 6 p.p..
The following F0 females were sacrificed by the same way without overnight fasting:
- females which did not deliver: on Day 25 or 26 p.c. (after a body weight recording to check for a possible un-noticed delivery).

GROSS NECROPSY
A macroscopic post-mortem examination of the principal thoracic and abdominal organs was performed on all F0 animals. This included examination of the external surfaces, all orifices, the cranial cavity, the external surfaces of the brain and spinal cord, the thoracic, abdominal and pelvic cavities with their associated organs and tissues and the neck with its associated organs and tissues. Special attention was paid to the reproductive organs. The numbers of corpora lutea and implantation sites were also recorded for females sacrificed as scheduled on Day 6 p.p. and for females sacrificed on Days 25 or 26 p.c. due to no delivery. For apparently non-pregnant female the presence of implantation scars on the uterus was checked.


HISTOPATHOLOGY / ORGAN WEIGHTS
A microscopic examination was performed on:
- all tissues listed in the Tissue Procedure Table (See Table 7.5.1/1 section 7.5.1 (Repeated dose toxicity (Rep. dose tox. oral V1 CiToxLAB 2013)) from the first five sacrificed as scheduled males and the first five females sacrificed on day 6 p.p. of the control and high-dose groups (groups 1 and 4),
- all macroscopic lesions of all groups.

Special emphasis was paid to the stages of spermatogenesis in the male gonads and histopathology of interstitial testicular cell structure.

ORGAN WEIGHTS: the body weight of each animal sacrificed as scheduled after the end of the mating period for males or on Day 6 p.p. for females was recorded before sacrifice and the organs specified in the Tissue Procedure Table (See Table 7.5.1/1 section 7.5.1 (Repeated dose toxicity (Rep. dose tox. oral V1 CiToxLAB 2013)) were weighed (wet) as soon as possible after dissection.
The ratio of organ weight to body weight (recorded immediately before sacrifice) was calculated.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring was sacrificed by an intraperitoneal injection of sodium pentobarbital on day 5 p.p..
- These animals were subjected to postmortem examinations (macroscopic examination)
Statistics:
The following statistical methods were used to analyze body weights, food consumption, reproduction and skeletal examination data:
• Means and standard deviations of various data were calculated.
• If the variables could be assumed to follow a normal distribution, the Dunnett t-test, based on a pooled variance estimate, was used for inter-group comparisons (i.e. single treatment groups against the control group).
• The Steel test (rank test) was applied when the data could not be assumed to follow a normal distribution.
• Fisher's Exact test for 2x2 tables was applied if the variables could be dichotomized without loss of information.
Reproductive indices:
- pre-implantation loss: Number of corpora lutea - Number of implantation sites/Number of corpora lutea x 100.

- post-implantation loss (manually calculated): Number of implantation sites - Number of live pups/Number of implantation sites x 100.

- mating index:Number of mated animals/Number of paired animals x 100.

- fertility index: Number of pregnant female partners/Number of mated pairs x 100.

- gestation index: Number of females with live born pups/Number of pregnant females x 100.


Offspring viability indices:
- live birth index: Number of live born pups/Number of delivered pups x 100.

- viability index on day 4 post-partum: Number of surviving pups on day 4 post-partum/Number of live born pups x 100.



Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
(Observed clinical signs were considered to be related to the test item but of minor toxicological significance)
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
(See details on results section)
Other effects:
not examined
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
There were no unscheduled deaths in males during the study.
For no delivery, one female given 30 mg/kg/day was sacrificed on Day 26 p.c. and two others given 100 mg/kg/day were sacrificed on Day 25 p.c.. These females were found to be non pregnant at necropsy.

Ptyalism was recorded in 6/10 males given 100 mg/kg/day and in all males and females given 300 mg/kg/day throughout the premating, gestation and/or lactation periods. This sign, commonly observed when a test item is administered by gavage, was considered to be of minor toxicological importance.
Other findings (area(s) of hair loss and cutaneous lesion(s) on neck), commonly observed in this species and strain, were considered to be not treatment related

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
There were no effects on mean body weight and mean body weight change in males and females during the study.
There were no effects on mean food consumption during the study in males and females.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
There were no test item-related effects on mating or fertility data.
The slightly high mean pre-coital time in control group was particularly due to one pair which mated after 13 days (the female was blocked in diestrous for several days).
All females were pregnant, except one female and two females at 30 and 100 mg/kg/day, respectively.

ORGAN WEIGHTS (PARENTAL ANIMALS)
When compared with controls, the following changes in the mean organ weights were observed:
- the absolute and relative liver weights were higher in males given 300 mg/kg/day but without reaching a statistical significance,
- the absolute and relative spleen weights were higher in males given 30 or 100 mg/kg/day reaching a statistical significance for the absolute weight at 100 mg/kg/day (p<0.05). In the absence of a similar trend at 300 mg/kg/day, any relationship with the test item was excluded,
- the absolute and relative adrenal weights were higher in males given 30 mg/kg/day without reaching a statistical significance. In the absence of a similar trend at 100 or 300 mg/kg/day, any relationship with the test item was excluded.

Other changes in the mean organ weights were considered to be part of the normal variation between groups.

GROSS PATHOLOGY (PARENTAL ANIMALS)
Thickening or white discoloration of the forestomach was seen in 1/10 males given 30 mg/kg/day, 1/10 males given 100 mg/kg/day, 9/10 males and 2/10 females given 300 mg/kg/day. This mainly correlated histologically with hyperplasia of squamous cells and/or hyperkeratosis and was considered to be related to the test item.

Black deposit was present in the stomach of one female given 100 mg/kg/day correlating histologically with erosion and one female given 300 mg/kg/day correlating with a chronic inflammation of the serosa with fibrosis. This stomach also appeared thickened. Any relationship with the test item was excluded.

Reduced size of testes was observed bilaterally in one male given 100 mg/kg/day (associated with reduced size of epididymides) and unilaterally in one male given 300 mg/kg/day (associated with agenesis of the epididymis). These isolated changes were considered to be fortuitous and without relationship with the test item.

Other macroscopic changes were considered to be part of the normal changes commonly seen in rats and were considered without any relationship with the test item.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Test item-related changes were seen in the forestomach at all dose-levels in males and at 100 or 300 mg/kg/day in females.
1- Forestomach:
The forestomach was examined histologically in the following animals:
- the first five sacrificed as scheduled males from all groups 1 to 4 (controls, 30, 100 and 300 mg/kg/day respectively) and those showing macroscopic changes at necropsy (one given 30 mg/kg/day and four given 300 mg/kg/day),
- the first five females sacrificed on Day 6 post-partum from groups 1 to 4 (controls, 30, 100 and 300 mg/kg/day respectively).

Hyperplasia of squamous cells (graded minimal to marked in males and minimal to moderate in females) was observed at all dose-levels in males and at 100 and 300 mg/kg/day in females. This was often associated with hyperkeratosis. Additional changes consisted of minimal erosion/ulceration in a few animals at 100 and 300 mg/kg/day and inflammation and/or edema

2- Miscellaneous:
Minimal mononuclear cell infiltrate (perivascular) was observed in lungs of all groups including controls, but with a higher incidence in treated animals than in controls. However, in the absence of a dose-related incidence and/or severity any relationship with the test item was excluded.

3- Testes:
Marked tubular atrophy was seen in the right testis of one male given 300 mg/kg/day correlating with the reduced size noted at necropsy.
Severe tubular atrophy was seen bilaterally in one male given 100 mg/kg/day associated with severe reduced sperm count in the epididymides correlating with the reduced size observed at necropsy.

As these changes are occasionally seen in rats, involved only one side in the high dose group male and in the absence of testes changes in the other treated animals, these isolated observations were considered to be fortuitous.

4- Liver
There were no histopathological correlates with the higher liver weight at necropsy in males at 300 mg/kg/day.

Dose descriptor:
NOAEL
Remarks:
(systemic toxicity)
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Based on absence of adverse findings at this high-dose level.
Dose descriptor:
NOEL
Remarks:
(Reproduction: mating and fertility)
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: There were no effects in the mating, fertility and gestation rates or in pre- and post-implantation losses.
Dose descriptor:
NOEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: There were no effects in pup mortality, clinical signs, sex ratio, mean body weight or macroscopic post-mortem findings.
Remarks on result:
other: Generation: progeny (migrated information)
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
There were no effects on pup viability.

CLINICAL SIGNS (OFFSPRING)
There were no test item-related effects on clinical signs (comparable incidences in control pups or not dose-related).

BODY WEIGHT (OFFSPRING)
There were no effects on mean body weight and mean body weight change in pups during the lactation period.

GROSS PATHOLOGY (OFFSPRING)
There were no test item-related macroscopic findings at pup necropsy.

OTHER FINDINGS (OFFSPRING): Pup sex ratio:
There were no test item-related effects on sex-ratios (% of male pups).
Reproductive effects observed:
not specified

No remarks.

Conclusions:
Under the test conditions of this study, the No Observed Adverse Effect Level (NOAEL) for parental toxicity (systemic) was considered to be 300 mg/kg bw/day based on absence of adverse findings at this high-dose level. No reprotoxic effects were observed in all treated animals. Therefore, the NOEL for reproduction, development and fertility was considered to be 300 mg/kg bw /day (the highest tested dose).
Executive summary:

A combined repeated dose toxicity study with the reproduction/developmental toxicity screening test (CiToxLAB, 2013) was conducted with Phosphoric acid, mono and bis(branched and linear pentyl) esters according to the OECD test guideline No. 422 and in compliance with GLP.

 

Sprague-Dawley rats (10/sex/dose level) received the test item daily by oral (gavage) administration before mating, through mating and, for the females, through gestation until day 5 p.p.. The test item was administered at dose levels of 30, 100 and 300 mg/kg/day in the vehicle (Corn oil). Another group of ten males and ten females received the vehicle, alone, under the same experimental conditions and acted as a control group. A constant dosage volume of 5 mL/kg bw/day was used.

 

The animals were checked daily for clinical signs, mortality, and detailed clinical observations were conducted weekly. Body weight and food consumption were recorded weekly until mating and then at designated intervals throughout gestation and lactation. A Functional Observation Battery (FOB), including motor activity, and laboratory investigations (haematology, blood biochemistry and urinalysis) were carried out on five males and five females from each group at the end of the study.

 

The males were sacrificed after completion of the mating period and dams were sacrificed on Day 6 p.p.. A full macroscopic post mortem examination was performed, with particular attention accorded to the reproductive organs. Designated organs were weighed and selected tissue specimens were preserved.

A microscopic examination was performed on selected organs from the first five males sacrificed at the end of the treatment period and the first five females sacrificed on Day 6 p.p. of the control and high-dose groups and on all macroscopic lesions from all groups.

A macroscopic post-mortem examination of the principal thoracic and abdominal organs was performed on pups sacrificed on Day 5 p.p. and on those found dead.

There were no unscheduled test item-related deaths. Ptyalism was the only recorded clinical sign, observed in both sexes at 300 mg/kg/day and in males at 100 mg/kg/day. This finding was considered to be related to the test item but of minor toxicological importance. There were no effects on body weight, food consumption, functional observation battery tests or motor activity data in any group and sex. No relevant changes were noted in haematology and biochemistry parameters. Mating, fertility and delivery data were unaffected by the test item treatment. No effects on pups were observed up to Day p.p. (viability, clinical signs, sex ratio, macroscopic post-mortem findings). At pathology examination, microscopic changes were seen in the forestomach at all dose-levels in males and at 100 and 300 mg/kg/day in females. Despite the absence of histopathological correlates, the higher liver weight in males at 300 mg/kg/day was considered to be test item-related but non adverse.

Based on the experimental conditions of this study:

-The No Observed Adverse Effect Level (NOAEL) for parental systemic toxicity was considered to be 300 g/kg/day based on absence of adverse findings at this high-dose level,

- The No Observed Adverse Effect Level (NOAEL) for local tolerance was considered to be lower than 30 mg/kg/day (pronounced microscopic changes in the forestomach in males from 30 mg/kg/day and in females from 100 mg/kg/day).

- The No Observed Effect Level (NOEL) for reproductive performance (mating and fertility) and toxic effects on progeny was considered to be 300 mg/kg bw/day based of absence of findings at this high-dose level.

 

This study is considered as acceptable as it satisfies the main criteria of OECD guideline No.422.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
300 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
OECD guideline 422 reprotoxicity screening study (Kr. 1).

Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

One study of reliability 1 according to Klimisch cotation critera, is available (CiToxLAB, 2013) and was selected as a key study.

 

In this study (OECD 422), Sprague-Dawley rats (10/sex/dose level) received the test item daily by oral (gavage) administration before mating, through mating and, for the females, through gestation until day 5 p.p.. The test item was administered at dose levels of 30, 100 and 300 mg/kg/day in the vehicle (Corn oil). Another group of ten males and ten females received the vehicle, alone, under the same experimental conditions and acted as a control group. A constant dosage volume of 5 mL/kg bw/day was used.

 

The animals were checked daily for clinical signs, mortality, and detailed clinical observations were conducted weekly. Body weight and food consumption were recorded weekly until mating and then at designated intervals throughout gestation and lactation. A Functional Observation Battery (FOB), including motor activity, and laboratory investigations (haematology, blood biochemistry and urinalysis) were carried out on five males and five females from each group at the end of the study.

 

The males were sacrificed after completion of the mating period and dams were sacrificed on Day 6 p.p.. A full macroscopic post mortem examination was performed, with particular attention accorded to the reproductive organs. Designated organs were weighed and selected tissue specimens were preserved.

A microscopic examination was performed on selected organs from the first five males sacrificed at the end of the treatment period and the first five females sacrificed on Day 6 p.p. of the control and high-dose groups and on all macroscopic lesions from all groups.

A macroscopic post-mortem examination of the principal thoracic and abdominal organs was performed on pups sacrificed on Day 5 p.p. and on those found dead.

There were no unscheduled test item-related deaths. Ptyalism was the only recorded clinical sign, observed in both sexes at 300 mg/kg/day and in males at 100 mg/kg/day. This finding was considered to be related to the test item but of minor toxicological importance. There were no effects on body weight, food consumption, functional observation battery tests or motor activity data in any group and sex. No relevant changes were noted in haematology and biochemistry parameters. Mating, fertility and delivery data were unaffected by the test item treatment. No effects on pups were observed up to Day p.p. (viability, clinical signs, sex ratio, macroscopic post-mortem findings). At pathology examination, microscopic changes were seen in the forestomach at all dose-levels in males and at 100 and 300 mg/kg/day in females. Despite the absence of histopathological correlates, the higher liver weight in males at 300 mg/kg/day was considered to be test item-related but non adverse.

Based on the experimental conditions of this study:

-The No Observed Adverse Effect Level (NOAEL) for parental systemic toxicity was considered to be 300 g/kg/day based on absence of adverse findings at this high-dose level,

- The No Observed Adverse Effect Level (NOAEL) for local tolerance was considered to be lower than 30 mg/kg/day (pronounced microscopic changes in the forestomach in males from 30 mg/kg/day and in females from 100 mg/kg/day).

- The No Observed Effect Level (NOEL) for reproductive performance (mating and fertility) and toxic effects on progeny was considered to be 300 mg/kg bw/day based of absence of findings at this high-dose level.


Short description of key information:
OECD 422 Guideline study:
The test item, Phosphoric acid, mono and bis(branched and linear pentyl)esters, was administered daily by oral gavage to male and female Sprague-Dawley rats, for 2 weeks before mating, during mating and (for females) throughout gestation and until Day 5 post-partum, at dose-levels of 30, 100 or 300 mg/kg/day.

Based on the experimental conditions of this study:
- the No Observed Adverse Effect Level (NOAEL) for local tolerance was considered to be lower than 30 mg/kg/day (pronounced microscopic changes in the forestomach in males from 30 mg/kg/day and in females from 100 mg/kg/day),
- the No Observed Adverse Effect Level (NOAEL) for parental systemic toxicity was considered to be 300 g/kg/day based on absence of adverse findings at this high-dose level,
- the No Observed Effect Level (NOEL) for reproductive performance (mating and fertility) and toxic effects on progeny was considered to be 300 mg/kg/day based of absence of findings at this high-dose level.

Justification for selection of Effect on fertility via oral route:
Only one study is available.

Effects on developmental toxicity

Description of key information
A testing proposal for an OECD 414 Guideline (Prenatal Developmental toxicity) study was submitted to ECHA.  
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no study available (further information necessary)
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information
Justification for selection of Effect on developmental toxicity: via oral route:
A testing proposal for an OECD Guideline 414 (Prenatal Developmental toxicity) study was submitted to ECHA.

Toxicity to reproduction: other studies

Additional information

Not applicable

Justification for classification or non-classification

- Effects on fertility:

Based on the available data, Phosphoric acid, mono-and bis (branched and linear pentyl) esters is not classified for fertility according to the Regulation (EC) No 1272/2008 and the Directive 67/548/EEC criteria.

- Effects on developmental toxicity:

No data was available; therefore no classification is possible due to lack of data. However, a testing proposal for an OECD 414 guideline study is submitted to ECHA.