[No public or meaningful name is available]

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Description of key information

Based on the results of an OECD 422  and GLP compliant study with the the read-across substance in rat, the following NOAELs were determined:

NOAELsystemic: 450 mg/kg bw/d

NOAELlocal: 200 mg/kg bw/d

NOAELreproduction: 450 mg/kg bw/d

NOAELdevelopment: 450 mg/kg bw/d

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

Please refer to read-across justification attached in IUCLID Section 13.

Reason / purpose for cross-reference:

read-across source

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

450 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Key result

Dose descriptor:

NOAEL

Remarks:

local

Effect level:

200 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

body weight and weight gain

food consumption and compound intake

gross pathology

histopathology: non-neoplastic

Key result

Dose descriptor:

NOAEL

Remarks:

reproduction

Effect level:

450 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Key result

Critical effects observed:

no

Key result

Dose descriptor:

NOAEL

Remarks:

development

Generation:

F1

Effect level:

450 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

450 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

GLP and guideline compliant study.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

OECD 422 with the read-across substance

The objective of this study was to obtain initial information on the toxic potential of test item and on the possible effects of the test item on reproduction and development when repeatedly administered orally (by gavage) to rats at doses of 80, 200 and 450 mg/kg bw/day compared to control animals according to OECD 422.

As a screening test, it was intended to provide initial information on the possible health hazards likely to arise from repeated exposure over a relatively limited period of time and on the possible effects on male and female reproductive performance such as gonadal function, mating behavior, conception, pregnancy, parturition as well as on development of the F1 offspring from conception to day 13 post-partum associated with administration of repeated maternal doses.

Phosphoric Acid, 2-ethylhexyl Ester, Sodium Salt was administered orally (by gavage) once daily at 0 (vehicle only), 80, 200 and 450 mg/kg body weight doses to four groups of Han:WIST rats consisting of 12 animals per sex per group in concentrations of 16, 40 and 90 mg/mL, corresponding to a 5 mL/kg bw dosing volume. A group of vehicle (5 % Tween 80 in distilled water) treated animals (n= 12/sex) served as a control.

The suitability of the chosen vehicle for the test item at the intended concentrations was analytically verified up front. Phosphoric Acid, 2-ethylhexyl Ester, Sodium Salt was stable in the vehicle in concentrations of 1 mg/mL and 200 mg/mL at room temperature for five days.

The concentration of the test item in the dosing formulations administered to the animals was checked two times during the study. Phosphoric Acid, 2-ethylhexyl Ester, Sodium Salt concentrations in the dosing formulations varied within the range of 96.3 % and 113 % in comparison to the nominal values confirming the proper preparation of the dosing formulations. All animals of the parent (P) generation were dosed prior to mating (14 days) and throughout mating. In addition, males received the test item or vehicle after mating up to the day before the necropsy (altogether for 49 days). Females were additionally exposed through the gestation period and up to lactation days 12-16, i.e. up to the day before necropsy (altogether for 53-56 days). Observations included mortality, clinical signs, body weight, food consumption, mating, pregnancy and delivery process, as well as development of offspring. Estrous cycle was monitored by examining vaginal smears before the treatment for two weeks and for two weeks from the beginning of the treatment period and during the mating period until evidence of copulation. Vaginal smears were prepared on the day of the necropsy for each dam. The dams were allowed to litter and rear their offspring up to day 13 post-partum. Litters were weighed and offspring were observed for possible abnormalities and were euthanized on post-natal day 13 or shortly thereafter. Blood samples were collected for possible determination of serum levels of thyroid hormones (FT4 and TSH) from 2-8 pups per litter (in litters with 9 or more pups) on post-natal day 4, from all dams and from 2-8 pups per litter at termination on post‑partum/post-natal day 13 and from all parent male animals and not mated, non-pregnant female animals at termination.

Five dams and their male mating partners were randomly selected from each group to examine further signs of toxicity such as functional observations, hematology and blood coagulation, clinical chemistry, gross necropsy, organ weighing and histopathology examination. All parental animals were subjected to gross pathology one day after the last treatment. The body weight, brain weight and weight of the testes and epididymides and prostate and seminal vesicles with coagulating glands as a whole of adult male animals were determined. In addition, for five males and females randomly selected from each group, adrenals, brain, heart, kidneys, liver, spleen and thymus were weighed.

Thyroid gland was preserved from all adult males and females and one male and one female pup per litter for the intended subsequent histopathological examination.

Histopathology examination was performed on ovaries, uterus with cervix and oviduct, vagina, testes, epididymides, prostate and seminal vesicles with coagulating gland in the control and high dose groups (male or female). These organs were processed and evaluated in non-pregnant female animal at 200 mg/kg bw/day.

Full histopathology examinations were performed on the preserved organs and tissues of the randomly selected animals in the control and high dose groups and in one male animal in control group with severe individual macroscopic findings in control group.

In addition, histopathological examinations were performed on the stomach of selected animals in the low and mid dose groups (on the basis of necropsy observation in high dose animals) and on organs showing macroscopic findings at the necropsy in animals in control, 80, 200 and 450 mg/kg bw/day groups.

 

There was no mortality in any groups (control, 80, 200 and 450 mg/kg bw/day).

Test item related clinical signs (decreased activity, narrow eye aperture, dyspnea or piloerection) were observed in male and female animals administered with 450 mg/kg bw/day with variable incidence and duration.

Nuzzling up the bedding material and salivation appeared in male animals at 80 mg/kg bw/day and in male and female animals at 200 and 450 mg/kg bw/day shortly after the administration and ceased within a short time period.

Some clinical signs (salivation before the treatment, piloerection, decreased activity or dyspnea) were sporadically detected at the detailed weekly clinical observation in male and female animals at 450 mg/kg bw/day.

The behavior and physical condition of the animals was normal at each dose level (80, 200 or 450 mg/kg bw/day) at the functional observations.

The body weight development was not adversely affected by the test item in male or in female animals at 80, 200 or 450 mg/kg bw/day during the entire treatment period (pre-mating and post-mating period for male animals; pre-mating, gestation and lactation periods for female animals).

Mean body weight gain at 450 mg/kg bw/day was reduced mainly during the pre-mating period, but had no influence on the overall body weight gain or on the mean body weight (male and female). Therefore, these findings were considered to be test item related, but non-adverse.

The mean daily food consumption was not adversely affected in male or female animals in control and at 80, 200 and 450 mg/kg bw/day during the entire study.

The slightly reduced mean food intake during the pre-mating period at 450 mg/kg bw/day correlated with the reduced body weight gain during this period. It was considered to be test item related, but non-adverse due to its minor degree.

A test item influence on the estrous cycle was not found at any dose level (80, 200 and 450 mg/kg bw/day).

There were no toxicologically relevant differences in the evaluated parameters of delivery data between the control and test item treated groups (80, 200 and 450 mg/kg bw/day). The litter size (mean number of births – total, live and viable) was slightly lowered with respect to the control in dams at 450 mg/kg bw/day. Although, the values met well the historical control ranges.

The fertility indices were reduced in male and female animals at 200 and 450 mg/kg bw/day. The individual and mean values met well the historical control ranges. In the lack of relevant changes (estrous cycle, pregnancy data, implantation sites, histopathology of sexual organs) lower mean fertility indices were considered to be insensitive factor in the determination of the NOAEL for reproductive performance

Hematological and blood coagulation investigation did not reveal adverse test item related changes in the examined parameters at 80, 200 or 450 mg/kg bw/day (male or female).

There were no test item related adverse effects on the examined clinical chemistry parameters at 80, 200 or 450 mg/kg bw/day (male or female).

There were no test item related changes in the serum thyroid hormone (FT4 and TSH) levels at any dose in parental males and FT4 levels in 13-day offspring. TSH levels were elevated in all test item groups in 13-day offsprings. As no dose response was seen, they were considered to be incidental and not related to test item treatment.

Macroscopic alterations related to the irritating effect of the test item were detected in the stomach (thickening of the mucous membrane) in a dose related manner in male and female animals at 200 and 450 mg/kg bw/day.

There were no adverse effects on the weights of examined organs in male or female animals at any dose level (80, 200 and 450 mg/kg bw/day). Minor changes in the weights of the liver (relative to body or brain weights) in male animals at 80, 200 and 450 mg/kg bw/day and in the weights of prostate, seminal vesicle with coagulating gland as a whole (absolute and relative to body and brain weights), at 450 mg/kg bw/day, were considered to be of little or no biological relevance in the lack of related histopathological changes.

Histopathological examinations did not reveal any toxic or other test item related lesions in the investigated reproductive organs of experimental male and female animals at 450 mg/kg bw/day.

Squamous cell hyperplasia with slight infiltration of inflammatory cells in the lamina propria in the mucous membrane of non-glandular (cardiac part of) stomach were observed in male and female animals at 200 and 450 mg/kg bw/day in a dose related manner.

The offspring’s development was undisturbed at 80, 200 and 450 mg/kg bw/day from birth to post-natal day 13. No adverse effects on the mortality, clinical signs, body weight, anogenital distance (male and female) or nipple retention (male) were detected.

 

Under the conditions of the present study, Phosphoric Acid, 2-ethylhexyl Ester, Sodium Salt administered at 80, 200 and 450 mg/kg bw/day by oral gavage did not adversely influence the reproductive performance (gonad function, mating behavior, parturition) in parental male and female Han:WIST rats in the absence of parental toxicity.

Test item influence on the slightly higher mean post-implantation (pre-natal) loss and lower mean birth per litter at 450 mg/kg bw/day might be presumed although values met well the historical control.

There were no clear signs of systemic toxicity in male or female animals at 80, 200 or 450 mg/kg bw/day. Transient clinical signs, minor changes in the body weight and food consumption seen at 450 mg/kg bw/day were considered as test item related, but non-adverse. Local effect was observed in the stomach at the necropsy (thickened mucous membrane at cardia) and histological examinations (squamous cell hyperplasia and inflammation) in male and female animals at 200 or 450 mg/kg bw/day. They can be ascribed to the irritating property of the test item.

There were no test item related findings in male or female animals at 80 mg/kg bw/day.

The development of the F1 offspring was not impaired from birth to post-natal day 13 after repeated oral administration of dams at 80, 200 or 450 mg/kg bw/day.

Based on these observations the No Observed Adverse Effect Levels (NOAEL) were determined as follows:

NOAEL for systemic toxicity of male/female rats:  450 mg/kg bw/day

NOAEL for local toxicity of male/female rats:         200 mg/kg bw/day

NOAEL for reproductive performance of male/ female rats: 450 mg/kg bw/day

NOAEL for F1 Offspring:    450 mg/kg bw/day

 

Effects on developmental toxicity

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on reproductive toxicity with a read-across substance, the registered substance does not require classification according to Regulation (EC) No 1272/2008 (CLP).