9-Octadecen-1-ol, (Z)-, phosphate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

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:

2013-01-24 to 2013-03-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and guideline compliance

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

, adopted 27th July 1995

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA Health Effects Test Guidelines, OPPTS 870.3550 Reproduction/Developmental Toxicity Screening Test, July 2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Species and strain: Hsd.Brl.Han:Wist rat
- Source: Toxi-Coop Zrt. Cserkesz u. 90. H-1103 Budapest Hungary
- Age at study initiation: 69-74 days old
- Weight at study initiation: (P) Males: 302-355 g; Females: 170-216 g;

- Fasting period before study:

- Housing: Before mating: 2 animals of the same sex/ cage; mating: 1 male and 1 female / cage; pregnant females will be housed individually. Males after mating: 2 animals / cage
- Diet: ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest, Germany ad libitum, and tap water from municipal supply, as for human consumption, from 500 mL bottles ad libitum.
- Water: drinking water
- Acclimatization time: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 - 70 %
- Air changes: 8-12 air exchanges/hour by central air-condition system.
- Photoperiod: 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Route of administration:

oral: gavage

Vehicle:

other: sunflower oil

Details on exposure:

The test item was formulated in the vehicle in concentrations of 200, 60 and 20 mg/mL. Formulations were prepared in the formulation laboratory of Test Facility beforehand and not longer than three days before the application.

Analytical verification of doses or concentrations:

yes

Frequency of treatment:

once per day

Remarks:

Doses / Concentrations:
100, 300, 1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

12 per sex and dose

Control animals:

yes, concurrent vehicle

Details on study design:

The dose setting was based on findings obtained in a previous 14-day oral gavage dose range finding study with Reaction product of oleyl alcohol with polyphosphoric acid in rats (Toxi-Coop study no. 559.400.3890) and was in agreement with the Sponsor. The high dose was chosen with the aim of inducing toxic effects but no mortality or severe suffering of animals. The low dose was chosen to induce no toxic effect.

Positive control:

not applicable

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Animals were inspected for signs of morbidity and mortality twice daily (at the beginning and end of each working day).
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: General clinical observations were made once a day, after treatment at approximately the same time, considering the peak period of anticipated effects after dosing. More detailed examinations were made weekly at the times of weekly weighing prior to and during the mating until necropsy. Observations were performed on the skin, fur, eyes and mucous membranes, autonomic activity (lachrymation, piloerection, pupil size, respiratory pattern, occurrence of secretions and excretions), circulatory and central nervous system, somatomotor activity and behavior pattern, changes in gait, posture and response to handling.

BODY WEIGHT: Yes
- Time schedule for examinations: All parental animals were weighed with an accuracy of 1 g. Parental males were weighed on the first day of dosing (day 0), at least weekly thereafter and at termination. Parental females were weighed on the first day of dosing (day 0) then weekly, on gestation days 0, 7, 14 and 21 and on post-partal days 0 (within 24 hours after parturition) and 4, as well as on the day of necropsy. Body weight of the female animals were additionally weighed on gestational days 10 and 17 in order to give accurate treatment volumes, but these data will not be evaluated statistically.

FOOD CONSUMPTION:
The food consumption was determined with a precision of 1 g weekly by reweighing the non-consumed diet during the treatment period (pre-mating, gestation days 0, 7, 14 and 21, lactation days 0 and 4) except during mating phase.

Oestrous cyclicity (parental animals):

In the female animals including not mated and non-pregnant animals, the ovaries had a normal structure characteristic for the species, age and phase of sexual cycle. The cortex contained primary, secondary and tertiary follicles and corpora lutea, indicating the active maturation of oocytes and ovulation. The epithelial capsule and ovarian stroma were normal in all cases as well.

Sperm parameters (parental animals):

In the male animals, the investigated organs of reproductive system (testes, epididymides) were histologically normal and characteristic for the sexually mature organism in all cases of control and treated groups. The various spermatogenic cells (the spermatogonia, the spermatocytes, the spermatids and spermatozoa), representing different phases in the development and differentiation of the spermatozoons and the interstitial cells were the same in quantity and morphology in the testes of the investigated animals. Histologically, epididymides were normal in all cases as well.

Litter observations:

Each litter was examined as soon as possible after delivery (within 24 h of parturition), to establish the number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than normal pups), and the presence of gross abnormalities.
Live pups were counted and, sexed, litters were weighed within 24 hours of parturition (on the day after parturition is complete), and on day 4 post-partum with an accuracy of 0.1 g.
In addition to the observations on parent animals, any abnormal behavior of the offspring was observed.
All litters were checked and recorded daily for the number of viable and dead pups. Dead pups found were subjected to necropsy by macroscopic examination. On day 0 of lactation, a lung flotation test was performed on all dead pups to separate stillborns from those that died after delivery. The lung flotation test is negative for stillborns (pups that died intrauterine) but positive for pups that died after delivery.

Postmortem examinations (parental animals):

Pathology
Gross necropsy was performed on each animal. One day after the last treatment, animals were euthanized by exsanguination after verification of the deep narcosis by Isofluran CP (details are presented in "Details of Other Materials").
After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed, any abnormality was recorded including details of the location, color, shape and size. Special attention was paid to the organs of the reproductive system. The number of implantation sites and of corpora lutea was recorded.
The testes, epididymides and brain of all male adult animals were weighed.
The uterus with cervix, vagina, testes, epididymides (total and cauda), prostate, and seminal vesicles with coagulating glands, ovaries, pituitary and all organs showing macroscopic lesions of all adult animals were preserved. Testes and epididymides were preserved in modified Davidson solution, all other organs in 4 % buffered formaldehyde solution.

Postmortem examinations (offspring):

No test item related macroscopic alterations were found in offspring subjected to gross pathological examination.

Statistics:

The statistical evaluation of appropriate data (marked †above) was performed with the statistical program package SPSS PC+4.0.
The homogeneity of variance between groups was checked by Bartlett’s homogeneity of variance test.
Where no significant heterogeneity was detected a one-way analysis of variance (ANOVA) was carried out. If the obtained result was significant, then Duncan Multiple Range test was used to assess the significance of inter-group differences. Getting significant result at Bartlett’s test the Kruskal-Wallis analysis of variance was used and the inter-group comparisons were performed using Mann-Whitney U-test. Chi2 test was performed, if feasible.
The frequency of clinical signs, pathology and histopathology findings were calculated.

Reproductive indices:

There were no differences between the control and test item treated groups (1000, 300 and 100 mg/kg bw/day) in the reproductive performance of male and female animals and in delivery data of dams.

Offspring viability indices:

Negative effects of the test item on offspring development (mortality, clinical signs, body weight and necropsy findings) were not detected between postnatal days 0 and 4.

Clinical signs:

no effects observed

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:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Mortality
There was no test item related mortality during the course of the study.

Clinical Observations

Daily Observations
Test item related salivation was observed at 1000 mg/kg bw/day (12/12 male and 12/12 female) and 300 mg/kg bw/day (3/12 male) from the first week of the study up to the termination. The behavior and physical condition of animals were considered to be normal.
Alopecia was noted for some female animals in all groups: 1/10 dams and 1/1 not mated female animal at 1000 mg/kg bw/day, 2/12 dams at 300 mg/kg bw/day, 3/11 at 100 mg/kg bw/day and 1/12 in the control group. Alopecia is a common dermal finding in this strain of experimental rats and was observed in all groups with similar incidence. Ttherefore, it had no toxicological meaning in this study.

Detailed Weekly Observations
There were no test item related clinical signs during the weekly detailed observations in male or female animals at any dose level during the entire observation period (pre-mating, mating, post-mating, gestation or lactation periods).
Alopecia as described above was observed in female animals of the control, 1000, 300 and 100 mg/kg bw/day groups.

Body Weight
The body weight development was not affected in male or female animals by the treatment with 1000, 300 or 100 mg/kg bw/day with respect to their controls.
Some statistically significant differences were noted for the higher mean body weight gain of male animals between Days 13 and 20 (300 mg/kg bw/day) and for the less mean body weight gain between Days 27 and 34 (male animals of 300 and 100 mg/kg bw/day groups).
Similarly, the mean body weight gain was slightly but statistically significantly higher than in the control group in female animals of 1000 mg/kg bw/day group between gestation days 0 and 7.
All these statistical significant differences with respect to control were not considered to be of toxicological significance.

Food Consumption
Test item related effects on the mean daily food consumption were not detected in male or female animals of the 1000, 300 or 100 mg/kg bw/day groups with respect to their controls.
The slightly less mean food consumption of male animals in the 100 mg/kg bw/day group between Days 7 and 13, Days 27 and 34, as well as between Days 34 and 41, were not judged to be toxicologically significant as similar findings were not observed in the higher dose groups. In the female animals of the 1000 mg/kg bw/day group, the mean daily food intake slightly exceeded the control value between pre-mating days 0 and 7 and between gestation days 7 and 14. However these slight differences were not considered to be of toxicological relevance.

Delivery Data of Dams
There were no test item related differences between the control and test item dosed groups in the delivery data of dams. The duration of pregnancy, the mean number of corpora lutea, implantation sites, pre-implantation loss, post-implantation loss and total intrauterine mortality was similar in the control and all test item treated groups. There were no significant differences between the control and test item treated groups in the mean number of total births, live-borns, stillborns and viable pups.

Reproductive Performance
There were no test item related differences between the control and test item treated male animals in the examined parameters of reproductive performance.
Statistical significances were noted for the higher percentage of not mated animals and less percentage of mated or sperm positive animals (i.e. copulatory index) in the 1000 mg/kg bw/day group (male and female) and higher percentage of non-pregnant females in 1000, 300 and 100 mg/kg bw/day groups. The percentage of fertile males and pregnant females, as well as the fertility indices (male and female) were slightly less in all test item treated groups (1000, 300 and 100 mg/kg bw/day) with respect to their control group. The gestation index was slightly less in the control group. However, all these statistically significant differences with respect to the appropriate control were judged to be toxicologically not relevant as all values were similar to the historical control data.



Necropsy
No test item related alterations were found at the macroscopic observations of organs and tissues at the necropsy in male and female animals at any dose level (1000, 300 and 100 mg/kg bw/day).



Organ Weight
The absolute and relative organ weights did not demonstrate any test item related alterations.
There were no significant differences between the control and test item treated groups in the examined organ weights (brain, testes and epididymides weights; absolute and relative to body and brain weights).


Histopathology
Histological examination of male and female genital organs (testes, epididymides and ovaries) did not reveal any toxic or other test item related alterations at 1000 mg/kg/bw/day dose.
In the male animals, the investigated organs of reproductive system (testes, epididymides) were histologically normal and characteristic for the sexually mature organism in all cases of control and treated groups. The various spermatogenic cells (the spermatogonia, the spermatocytes, the spermatids and spermatozoa), representing different phases in the development and differentiation of the spermatozoons and the interstitial cells were the same in quantity and morphology in the testes of the investigated animals. Histologically, epididymides were normal in all cases as well.
In the female animals including not mated and non-pregnant animals, the ovaries had a normal structure characteristic for the species, age and phase of sexual cycle. The cortex contained primary, secondary and tertiary follicles and corpora lutea, indicating the active maturation of oocytes and ovulation. The epithelial capsule and ovarian stroma were normal in all cases as well.
In one not mated female animal (1/1 of 1000 mg/kg bw/day group) a follicular cyst and in a non- pregnant female of the 100 mg/kg bw/day group (1/1) lack of corpora lutea in the ovary (one side) were observed. These findings – without inflammatory or other pathological lesions – are a slight neuro-hormonal phenomenon and could be considered as individual physiological disorders without pathological significance. It should be noted however, that these findings could explain the temporary infertility of the affected individuals.

Dose descriptor:

NOAEL

Remarks:

for general toxicity

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: no effects

Dose descriptor:

NOAEL

Remarks:

for reproductive performance

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: no effects

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

Mortality
There was no test item effect on extra uterine mortality of offspring.
The number and percentage of liveborns and viable offspring, extra uterine mortality were similar in the control and all test item treated groups between postnatal days 0 and 4.

Sex Distribution
There were no significant differences between control and test item treated groups in the ratio or in the litter means of genders on postnatal days 0 or 4.


Clinical Observations
Test item related clinical signs did not appear in the pups.


Body Weight
The mean litter weight and mean pup weight were similar in the control and test item treated groups, no test item related changes were found.

Necropsy
No test item related macroscopic alterations were found in offspring subjected to gross pathological examination.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: no effects

Reproductive effects observed:

not specified

Conclusions:

Under the conditions of the present study Reaction Product of Oleyl Alcohol with Polyphosphoric Acid did not cause toxic changes and did not influence reproductive performance (gonad function, mating behavior, conception, pregnancy, parturition) in parental male and female Hsd.Brl.Han: Wistar rats or development of the F1 offspring from conception to day 4 post-partum after repeated dose oral administration of 1000, 300 or 100 mg/kg bw/day.

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

NOAEL for male rats: 1000 mg/kg bw/day
NOAEL for female rats: 1000 mg/kg bw/day

NOAEL for reproductive performance of the male rats: 1000 mg/kg bw/day
NOAEL for reproductive performance of the female rats: 1000 mg/kg bw/day

NOAEL for F1 Offspring: 1000 mg/kg bw/day

Executive summary:

Four groups of Hsd.Brl.Han:Wist rats (n=12/sex/group) were administered orally (by gavage) once a day at 0 (vehicle only),1000, 300 and 100 mg/kg bw/dayat concentrations of 0, 200, 60 and 20 mg/mL corresponding to 5 mL/kg bw dose volume.

The suitability of the chosen vehicle for the test item at the intended concentrations was analytically verified up front. Reaction Product of Oleyl Alcohol with Polyphosphoric Acid in sunflower oil was stable for 24 hours at room temperature and for three days in the refrigerator.

Concentration of the test item in the dosing formulations varied from 91 % to 104 % of nominal concentrations and samples proved to be homogenous at both analytical occasions, thereby confirming proper dosing.

All animals of the parent (P) generation received test item or vehicle prior to mating (14 days) and throughout mating. Test item or vehicle was administered to male animals post mating (altogether for 41 days) up to the day before the necropsy. For dams, test item was administered through the gestation period and up to lactation day 3, 4 or 5 (altogether for 41 – 46 days), i.e. up to the day before the necropsy.

Observations included mortality, clinical signs, body weight, food consumption, mating, pregnancy and delivery process, as well as development of pups. The dams were allowed to litter, and rear their young up to termination on days 4 postpartum. Pups were weighed and observed for possible abnormalities. All parental animals were subjected to gross pathology one day after the last treatment. Histopathology examination was performed on reproductive organs (testes, epididymides and ovaries) in the control and high dose groups. The reproductive organs of non-pregnant females and males cohabited with in the low and middle dose groups were also processed and evaluated histologically.

Under the conditions of the present study Reaction Product of Oleyl Alcohol with Polyphosphoric Acid did not cause toxic changes and did not influence reproductive performance (gonad function, mating behavior, conception, pregnancy, parturition) in parental male and female Hsd.Brl.Han: Wistar rats or development of the F1 offspring from conception to day 4 post-partum after repeated dose oral administration of 1000, 300 or 100 mg/kg bw/day. Based on these observations the No Observed Adverse Effect Levels (NOAEL) were determined as follows: NOAEL for male rats: 1000 mg/kg bw/day NOAEL for female rats:1000 mg/kg bw/day NOAEL for reproductive performance of the male rats:1000 mg/kg bw/day NOAEL for reproductive performance of the female rats: 1000 mg/kg bw/day NOAEL for F1 Offspring:1000 mg/kg bw/day

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The study is reliable based on its GLP and guideline compliance.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Short description of key information:
Under the conditions of a Reproduction/Developmental Toxicity Screening Test according to OECD guideline 421 the Reaction Product of Oleyl Alcohol with Polyphosphoric Acid did not cause toxic changes and did not influence reproductive performance (gonad function, mating behavior, conception, pregnancy, parturition) in parental male and female Hsd.Brl.Han: Wistar rats or development of the F1 offspring from conception to day 4 post-partum after repeated dose oral administration of 1000, 300 or 100 mg/kg bw/day.

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

NOAEL for male rats: 1000 mg/kg bw/day
NOAEL for female rats: 1000 mg/kg bw/day

NOAEL for reproductive performance of the male rats: 1000 mg/kg bw/day
NOAEL for reproductive performance of the female rats: 1000 mg/kg bw/day

NOAEL for F1 Offspring: 1000 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

Based on the results of a Reproduction/Developmental Toxicity Screening Test the test item is not subject to classification as reproductive toxic according to the criteria of EU Directive 67/548/EEC and EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No 1272/2008.