Tridecan-1-ol

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Description of key information

Reproductive toxicity study

Based on the data available from different studies, NOAEL for test material was considered to be 1000 mg/kg Bwt/day .When male and female rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Link to relevant study records

Reference

Reference 1

Endpoint:

extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)

Type of information:

experimental study planned

Study period:

once approvied by ECHA

Justification for type of information:

CONSIDERATIONS THAT THE GENERAL ADAPTATION POSSIBILITIES OF ANNEX XI OF THE REACH REGULATION ARE NOT ADEQUATE TO GENERATE THE NECESSARY INFORMATION:
- Available GLP studies: not available
- Available non-GLP studies: not available
- Historical human data: not available
- (Q)SAR: not available
- In vitro methods: not available
- Weight of evidence: not available
- Grouping and read-across: not available
-Substance-tailored exposure driven testing: Not applicable

1. We intend to upgrade our tonnage band from 1-10MT to 1000+MT as JS member wants to join in 1000+MT, which means that REACH Annex X requirements are applicable to our registration. As per the tonnage band requirement, we are willing to perform the testing for the endpoint toxicity to reproduction extended one generation reproductive toxicity study (OECD 443).

2. Another reason for proposing to conduct this study in accordance with Annex X, is that there is insufficient information available for repeated (28 day and 90 days) toxicity studies and reproductive toxicity studies for the registered substance to warrant adequate classification. Thus, by performing this study we will have a better understanding of the concerns regarding the potential for adverse effects on fatality or development.

Qualifier:

according to guideline

Guideline:

OECD Guideline 443 (Extended One-Generation Reproductive Toxicity Study)

GLP compliance:

yes

Justification for study design:

Extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
TESTING PROPOSAL ON VERTEBRATE ANIMALS
1) Premating exposure duration: 10 weeks
The standard ten week premating exposure duration is proposed for this study.
2) Route of exposure : oral

Route
We propose this study be done by the oral route even though this is an expected route of exposure to this substance. The oral route is preferred because it will further increase the bioavailability of the test substance to test animals.

3) Extension of Cohort 1B and termination time for F2: extension not justified

According to column 2 (specific rules for adaptation from column 1) point 8.7.3 of the amended REACH Annex X, extension of cohort 1B to include the F2 generation shall be proposed by the registrant based on the following conditions being met (a and any of b(i), b(ii) or b(iii)) as shown in table 1 below (See also: Chapter R.7a: Endpoint sepcific guidance Version 4.0 - July 2015)):
Table 1: Basis for proposing an extension of cohort 1B to the F2 generation
A. The substance has uses leading to significant exposure of consumers or professionals, taking into account, inter alia, consumer exposure from articles Yes – The substances have uses not leading to significant exposure of consumers or professionals
B (i). The substance displays genotoxic effects in somatic cell mutagenicity tests in vivo which could lead to classifying it as Mutagen Category 2, or No – The substances are not classified as Mutagen Category 1A or 1B or 2.

B (ii). There are indications that the internal dose for the substance and/or any of its metabolites will reach a steady state in the test animals only after an extended exposure, or No – The substances are not classified as a PBT or vPvB. The log Pow is very low, the BCF were also very low, BCF of less than 2000 for substance were predicted from USEPA EPI Suite model. The toxicokinetic behaviour of the substances gives no hints for very slow clearance. Therefore, there are no indications that the internal dose for the substance and/or any of its metabolites will reach a steady state in the test animals only after an extended exposure.

B (iii) There are indications of one or more relevant modes of action related to endocrine disruption from available in vivo studies or non-animal approaches YES - There are no indications based on the available study results that endocrine disruption is a relevant mode of action for the substances, but additionally yes structural alerts exist.

Therefore, based on the above considerations, the registrant does believe that there is a basis for extending cohort 1B and not to include the F2 generation.

Additionally, the registrant is of the opinion that the extension of the cohort to F2 generation is unlikely to provide any meaningful results beyond those that will be obtained from the F1 generation in the proposed study.

4) Inclusion of Cohorts 2A and 2B: not justified

The registrant does not believe there is a need to include cohorts 2A and 2B in the test design. This is based on:
• Data from previous studies with the substance do not indicate neurotoxic effects such as changes in brain weight or in specific neural areas not secondary to body weight, changes in brain volume or specific neural areas or (histo)pathological findings in brain, spinal cord and/or nerves
• test animals exposure to the substance have not expressed any behavioural changes in the absence of general toxicity
• the substance is not known to have any mode of action associated with neurotoxicity such as cholinesterase inhibition and thyroid toxicity

5) Inclusion of Cohort 3: not justified

The registrant does not believe there is a need to include cohort 3 in the test design. This is based on:
• the substance has not caused biologically significant changes in haematology/clinical chemistry and/or organ weight associated with immunotoxicity such as reduced leucocyte count in combination with reduced spleen weight in repeated dose studies
• the substance has not caused significant effects to immunology organs such as thymus atrophy in repeated dose studies
• the substance is not classified as a (respiratory) sensitizer
• there are no indications that endocrine disruption is a relevant mode of action for the substance
• no structural analogues are known to show immunotoxic effects

NON-CONFIDENTIAL NAME OF SUBSTANCE:
- Name of the substance on which testing is proposed to be carried out:

CONSIDERATIONS THAT THE GENERAL ADAPTATION POSSIBILITIES OF ANNEX XI OF THE REACH REGULATION ARE NOT ADEQUATE TO GENERATE THE NECESSARY INFORMATION [please address all points below]:
- Available GLP studies: not available
- Available non-GLP studies: not available
- Historical human data: not available
- (Q)SAR: not available
- In vitro methods: not available
- Weight of evidence: not available
- Grouping and read-across: not available


CONSIDERATIONS THAT THE SPECIFIC ADAPTATION POSSIBILITIES OF ANNEXES VI TO X (AND COLUMN 2 THEREOF) OF THE REACH REGULATION ARE NOT ADEQUATE TO GENERATE THE NECESSARY INFORMATION:
- response of ECHA

Species:

rat

Strain:

Wistar

Sex:

male/female

Reference 2

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental data from various test chemicals

Justification for type of information:

Weight of evidence approach based on the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on reproductive toxicity studies on rats

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data available

Species:

rat

Strain:

other: 1.COBS CD 2.Crj: CD(SD)3.Wistar

Details on species / strain selection:

No data available

Sex:

female

Details on test animals or test system and environmental conditions:

2.Details on test animals and env. conditions
TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: 8 weeks old
- Weight at study initiation: male 312.1 to 363.7 g, female 205.3 -230.8 g).
- Fasting period before study:
- Housing: individually housed in a wire mesh floor cage (Nihon Cage ), raised, and allowed to take solid feed
(CE - 2, Japan Clea )
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period:7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 1 ° C.
- Humidity (%):50 to 65%,
- Air changes (per hr): about 15 times / hour
- Photoperiod (hrs dark / hrs light): a lighting condition set at 12 hours of lighting (7 am to 7 pm)
3.Details on test animal & Environmental conditions
TEST ANIMALS
- Source: Thomae GmbH, Biberach, Germany
- Age at study initiation: 68-85 days old
- Weight at study initiation: mean weight was approximately 214-233 g.
- Fasting period before study: No data available
- Housing: the rats were housed singly
in type DK III stainless-steel wire-mesh cages (Becker
and Co., Castrop-Rauxel, Germany) (floor area
about 800 cm2)
- Diet (e.g. ad libitum): ): Food ad libitum (The food
during the study was ground Kliba feed 343
rat/mouse/hamster (Klingentalmiihle AG, Kaiseraugst,
Switzerland).
- Water (e.g. ad libitum):water available ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%):30-70%.
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): The 12-hr light/dark cycle extended from 6.00 am to 18.00 pm and from 18.00 pm to 6.00 am.

Route of administration:

oral: gavage

Vehicle:

other: 1&2 corn oil 3.doubly-distilled water containing approximately 0.005% Cremophor EL".

Details on exposure:

1.PREPARATION OF DOSING SOLUTIONS:
Test material diluted with corn oil
2.Details on exposure
PREPARATION OF DOSING SOLUTIONS:
The test substance was prepared by suspending it in corn oil and used as a test sample
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil
- Concentration in vehicle: 0 ,100, 300 and 1,000 mg/kg bw/day
- Amount of vehicle (if gavage): 5ml/kg bw
- Lot/batch no. (if required): (Lot No. V 6 H 2050, Nacalai Tesque ),
- Purity: No data available

3.Details on oral exposure
PREPARATION OF DOSING SOLUTIONS:
The chemicals were freshly prepared for gavage administration every day in aqueous emulsions under rapid stirring in doubly-distilled water containing approximately 0.005% Cremophor EL".

VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0,130,650,975,1300mg/kg day
- Amount of vehicle (if gavage): 5 ml/kg body weight
- Lot/batch no. (if required):
- Purity:

Details on mating procedure:

2.
- M/F ratio per cage: 1/1
- Length of cohabitation: at the most 14days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy : until proof of pregnancy (formation of vaginal closing or sperm detection in vagina)
3.- M/F ratio per cage:1:4(one to four untreated females were mated with one untreated fertile male of the same breed)
- Length of cohabitation: Mating took place overnight
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:The presence of a dropped copulation plug was considered evidence of successful mating, and designated as gestation day (gd) 0
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- After successful mating each pregnant female was caged (how):
- Any other deviations from standard protocol:

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Study 1
10 days (from days 6-15 of gestation)
Study 2
Males: 42 days
Females: from 14 days prior to mating to day 3 of lactation
Study 3.
10 days (from day 6 to day 15 post-coitum (pc))

Frequency of treatment:

Daily

Details on study schedule:

No data available

Remarks:

Study 1
0,200,1000mg/kg bw
Study 2.
0 (vehicle), 100, 300 or 1000 mg/kg/day.
Study 3.
0,130,650,975,1300mg/kg day

No. of animals per sex per dose:

1.
Total:75
0 mg/kgbw/day:25 female
200 mg/kgbw/day:25 female
1000 mg/kgbw/day:25 female
2.
Total :104
0 (Vehicle): 13 male and 13 female
100 mg/kg/day: 13 male and 13 female
300 mg/kg/day: 13 male and 13 female
1000 mg/kg/day: 13 male and 13 female
3.
Total:45-60
0 mg/kgbw/day:8-10 female
0 mg/kgbw/day:8-10 female
130 mg/kgbw/day: 8-10 female
650 mg/kgbw/day: 8-10 female
975 mg/kgbw/day: 8-10 female
1300mg/kgbw/day:8-10 female

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

No data available

Parental animals: Observations and examinations:

1.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: Yes
- Time schedule : daily
- Cage side observations checked in table [No.?] were included. No data available

DETAILED CLINICAL OBSERVATIONS: No data available
- Time schedule: No data available

BODY WEIGHT: Yes
- Time schedule for examinations:
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: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
- Time schedule for examinations: No data available

OTHER: No data available
2.
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS:

Time schedule: Daily

BODY WEIGHT: Yes
Time schedule for examinations: For all males, measurements were made on 1, 8, and 15 days of administration 1 (administration start date), 8, 15,
22, 29, 36, 42 days and postmenopausal days and females for all cases,
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): yes For all males, measurements were made on 1, 8, and 15 days of administration (administration start date), 8, 15,22, 29, 36, 42 days and postmenopausal days and females for all cases,


Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
Time schedule for examinations
3.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: Yes
- Time schedule : daily
- Cage side observations checked in table [No.?] were included. No data available

DETAILED CLINICAL OBSERVATIONS: No data available
- Time schedule: No data available

BODY WEIGHT: Yes
- Time schedule for examinations: daily
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): daily
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
- Time schedule for examinations: No data available

Oestrous cyclicity (parental animals):

2.Estrous cycle length and pattern not performed because the test was Conducted by the TG adopted in 1990.

Sperm parameters (parental animals):

2.Sperm examination were not performed because the test was Conducted by the TG adopted in 1990.

Litter observations:

1.Intrauterine survival, foetal weight and external, skeletal and visceral anomalies were recorded.
2.litter size at birth, neonatal death, sex ratio of pups were observed

Postmortem examinations (parental animals):

1.Post-mortem examinations (Parent Animal)
SACRIFICE
The dams were sacrificed and sectioned on gestation day 20
GROSS NECROPSY:yes
2.Postmortem examinations (Parent Animal)
SACRIFICE : males were killing under pentobarbital sodium deep anesthesia the next day (day corresponding to 43 days of administration). Females delivered were sacrificed on the fourth day of nursing, females that did not deliver to the 25th day of pregnancy, females who did not copulate were exsanguinated and lethal under pentobarbital sodium anesthesia at the end of the mating period,
GROSS NECROPSY: yes
HISTOPATHOLOGY / ORGAN WEIGHTS: yes
Weights of the heart, liver, kidney, thymus, testis and epididymis were measured for all cases ,the testes and epididymis were fixed and stored in Bouin's solution, and other organs and brain, spleen, adrenal gland and bladder were fixed and fixed in 10% formalin. For these high-dose groups and control groups, paraffin sections were prepared according to a conventional method, and hematoxylineosin staining was performed to perform histopathological examination.
3.Post-mortem examinations (Parent Animal)
SACRIFICE
On day 20 pc all surviving females were killed
GROSS NECROPSY:
The uterus and ovaries were removed and the following data recorded: weight of uterus before opening;
number of corpora lutea; number of implantations classified as:
live foetuses, dead implantations, early resorptions, using the staining method according to Salewski (1964), early and late resorptions ,dead foetuses. Conception rates, and pre- and postimplantation losses were calculaled

Postmortem examinations (offspring):

1.SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- foetal weight and external, skeletal and visceral anomalies were recorded.

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
2.SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
3.The foetuses were dissected from the uterus, weighed and further investigated for external, visceral and skeletal findings. Approximately one-half of the foetuses of all grorLps was fixed in Bouin's solution and examined according to the method of Barrow and Taylor (1969). For skeletal findings approximately one-half of the foetuses was fixed in ethyl alcohol and stained according to a modified method of Dawson (1926)

Statistics:

2.Dunnett’s or Scheffe’s test for continuous data, Chi square test for copulated index and fertility index, and Mann-Whitney U test or Fisher’s test for histopathological examination data.
3.The data were evaluated using Dunnett's test (Dunnett, 1955 and 1964) for evaluation of food consumption, body weight, body weight change and corrected body weight gain (net maternal body weight change), weight of the uterus before opening, placental and foetal weights, and the number of corpora lutea, implantations, pre- and postimplantation losses, resorptions, and live and dead foetuses. Fisher's exact test (Dixon, 1981) was used for evaluation of the conception rate, mortality (of the dams) and all foetal findings

Reproductive indices:

No data available

Offspring viability indices:

No data available

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

1.clinical signs of intoxication were observed
2.In males, no abnormalities in general conditions were observed. In females, there was only one change from day 42 of administration (day of delivery) to the one in 100 mg / kg administration group (administration 42 days: piloerection, blood-like fluid discharge from the vaginal opening; administration 43 to 44 days: coat contamination) But it recovered on day 45 of administration. In other females, no abnormalities in the general condition were observed.
3.Lateral and abdominal position, unsteady gait, salivation, piloerection, nasal discharge and pneumonia was observed at 650,975,1300mg/kg day dose group.

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Description (incidence):

2.No deaths in male and female was observed.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

1.reduced body weight was observed at 1000mg/kg bw dose group
2.In male, the body weight gain increased from 8 to 15 days in the 100 mg / kg administration group significantly (p <0.01) compared with the control group, However, in the group administered with 300 mg / kg or more, there was no significant difference between the control group and treated group. In females, there was no significant difference in body weight and body weight gain between control group and treated group before breeding, pregnancy period and nursing period.
3.A slight decrease in body weight gain was observed at 650,975,1300mg/kg day dose group

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

A slight decrease in food consumption was observed at 650,975,1300mg/kg day dose group

Food efficiency:

not specified

Description (incidence and severity):

2.There was no significant difference between the control group and treated group for males and female at any time of feeding intake

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Atrophy of seminiferous tubules was found in 2 animals in the 1000 mg / kg administration group, one on both sides and one on one side, but both were localized and extremely mildly changed. There were no other abnormalities. Sperm granuloma was observed in one of the control group, and there was no abnormality. There were no abnormalities in the ovaries of the unexpanded and infertile cases.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

There was no significant difference between the control group and treated group in the mating rate, conception rate, the number of days taken from the start of cohabitation to mating and the number of estrus periods recurred during that period.
An abnormality in labor condition (not treated with placenta) and abnormality in nursing condition (not collecting children, poor protrusion of nipple, poor protrusion of infant, child's fetus) was administered to one of 100 mg / kg administration group, The decrease in body surface temperature) was observed, and all children died on nursing day 2, but no abnormality was observed in other mother animals.
Birth rate was 100% in all administration groups, and no significant difference was observed between gestation period between control group and treated group. No significant difference was observed between the control group and treated group for the number of luteinism, the number of implantation and the implantation rate of pregnant animals.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

clinical signs

body weight and weight gain

reproductive performance

Remarks on result:

other: overall no toxic effects was observed

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

2.There was no significant difference between the control group and treated group for the number of births, delivery rate, birth rate, fertility rate, and survival rate on 4th day of newborn.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

2.There was no significant difference between the control group and the treated group on body weight at 0 and 4 days of nursing

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Description (incidence and severity):

1.Effects on the foetus were confined to an increase in numbers of litters with the skeletal variant 'malaligned sternebrae' which occured at 200 mg/kg/day only and a slight decrease in foetal weight at 1000 mg/kg/day which was within the historical control range. As an increased incidence of 'malaligned sternebrae' was not observed at 1000 mg/kg/day the observation at 200 mg/kg/day was considered incidental.
2.No abnormalities were found in all pups examined in either the external observation at day 0 or the autopsy performed at day 4 after birth.
3.All foetal values were within the range of biological variation; any differences in malformations and retardations were statistically insignificant or without a dose-response relationship and mostly occurred in animals of the treated and untreated groups as well as the historical control group at comparable frequency. A single cheiloschisis and one anophthalmy in the top-dose group was considered coincidental and not biologically significant

Histopathological findings:

not specified

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

gross pathology

Remarks on result:

other: No developmental toxic effects was observed

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Reproductive effects observed:

not specified

Treatment related:

not specified

Conclusions:

The no observed adverse effect level (NOAEL) was considered to be 1000mg/kg bw/day for reproductive and developmental toxicity .When male and female rats were treated with test material by orally.

Executive summary:

Data available from different studies were reviewed to determine the reproductive toxicity of test chemical.The studies are as mentioned below:

Study 1.

The reproductive and developmental toxicity study of test material was performed in femaleCOBS CD rats. The test material was dissolved in corn oil and administered in dose concentration 0,200,1000mg/kg bw by oral gavage route from days 6-15 of gestation. 25 females /dose group were used in study. All the animals were observed forClinical signs, body weight and food consumoption.The dams were sacrificed and sectioned on gestation day 20. Intrauterine survival, foetal weight and external, skeletal and visceral anomalies were recorded. In dams clinical signs of intoxication were observed also reduced body weight was observed at 1000mg/kg bw dose group. Effects on the foetus were confined to an increase in numbers of litters with the skeletal variant 'malaligned sternebrae' which occurred at 200 mg/kg/day only and a slight decrease in foetal weight at 1000 mg/kg/day which was within the historical control range. As an increased incidence of 'malaligned sternebrae' was not observed at 1000 mg/kg/day the observation at 200 mg/kg/day was considered incidental. Hencethe no observed adverse effect level (NOAEL) was considered to be 1000mg/kg bw/day for reproductive and developmental toxicity .When female rats were treated with test material by orally during gestation days 6-15.

Study 2.

Combined Repeat Dose and Reproductive / Developmental Toxicity Screening Test of test material were performed on male and female Sprague Dawley rats. The test material was suspended in corn oil in dose concentration0 ,100, 300 and 1,000 mg/kg bw/day and administered via oral gavage route in dose volume 5ml/kg bw. Dose selected on the bases ofpreliminary test by the previously repeated 14-day oral administration. 13 male and 13 female each were placed in each group. Administration period For each males, 2 weeks before mating, and 42 days from the end of the mating period to the day before the necropsy for males, and for female 2 weeks before mating and a maximum of 2 weeks mating period (Until mating) and once daily for the entire gestation period and 3 days after nursing (delivery day = nursing 0 day) in mating females. Matings were made with the same sex of the same group living together within the same group for up to two weeks from the evening on 15th day of administration. Mating was confirmed every morning by examining the presence of sperm in the vaginal plug and vaginal smear and females confirmed to be mated were separated from male starting from that day as the 0th day of pregnancy and raised individually. All the animals were observed forClinical signs, body weight and food consumption.

No deaths in male and female were observed. In males, no abnormalities in general conditions were observed. In females, there was only one change from day 42 of administration (day of delivery) to the one in 100 mg / kg administration group (administration 42 days: piloerection, blood-like fluid discharge from the vaginal opening; administration 43 to 44 days: coat contamination) But it recovered on day 45 of administration. In other females, no abnormalities in the general condition were observed. In male, the body weight gain increased from 8 to 15 days in the 100 mg / kg administration group significantly (p <0.01) compared with the control group, However, in the group administered with 300 mg / kg or more, there was no significant difference between the control group and treated group. In females, there was no significant difference in body weight and body weight gain between control group and treated group before breeding, pregnancy period and nursing period. There was no significant difference between the control group and treated group for males and female at any time of feeding intake. There was no significant difference between the control group and treated group in the mating rate, conception rate, the number of days taken from the start of cohabitation to mating and the number of estrus periods recurred during that period.

An abnormality in labor condition (not treated with placenta) and abnormality in nursing condition (not collecting children, poor protrusion of nipple, poor protrusion of infant, child's fetus) was administered to one of 100 mg / kg administration group, The decrease in body surface temperature) was observed, and all children died on nursing day 2, but no abnormality was observed in other mother animals. For male In the 100 mg / kg administration group, the actual weight and the specific body weight value of the liver increased significantly (p <0.05) compared to the control group, but not the dose-dependent change. Atrophy of seminiferous tubules was found in 2 animals in the 1000 mg / kg administration group, one on both sides and one on one side, but both were localized and extremely mildly changed. There were no other abnormalities. Sperm granuloma was observed in one of the control group, and there was no abnormality. There were no abnormalities in the ovaries of the unexpanded and infertile cases. For female In the 100 mg / kg administration group, the actual weight of the kidney decreased significantly (p <0.05) compared to the control group, but not the dose-dependent change. For other organs in male and female, no significant difference was observed between the control group and treated group. Birth rate was 100% in all administration groups, and no significant difference was observed between gestation period between control group and treated group. No significant difference was observed between the control group and treated group for the number of luteinism, the number of implantation and the implantation rate of pregnant animals.There was no significant difference between the control group and treated group for the number of births, delivery rate, birth rate, fertility rate, and survival rate on 4th day of newborn. In addition, no significant difference was observed between the control group and treated group for sex ratio.There was no significant difference between the control group and thetreated grouponbody weight at 0 and 4 days of nursing. Births that showed morphological abnormalities were not observed in the external table observation on thebirthday, necropsy of the dead child, and necropsy on the 4th day of nursing. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 1000mg/kg bw .When male and female Sprague Dawley rats were treated with test material orally.

 Study 3.

The reproductive and developmental toxicity study of test material was performed on female wistar rats.The chemicals were freshly prepared for gavage administration every day in aqueous emulsions under rapid stirring in doubly-distilled water containing approximately 0.005% Cremophor EL". The test material in dose concentration0,130,650,975,1300mg/kg day was adminstered fromday 6 to day 15 post-coitum (pc). Two control groups, treated with either doubly-distilled water alone (1) or water plus approximately 0.005% Cremophor EL as emulsifier (2), were employed.one to four untreated females were mated with one untreated fertile male of the same breed. Mating took place overnight. If sperm was detected microscopically in the vaginal smears in the morning, the animals were considered to be fertilized. This day was designated as 'day 0' (beginning of the study) and the following day 'day 1' post-coitum (pc).Food consumption, body weights and clinical signs were recorded daily throughout the study. On day 20 pc all surviving females were killed and subjected to gross pathology.The foetuses were dissected from the uterus, weighed and further investigated for external, visceral and skeletal findings. Approximately one-half of the foetuses of all groups was fixed in Bouin's solution and examined according to the method of Barrow and Taylor (1969). For skeletal findings approximately one-half of the foetuses was fixed in ethyl alcohol and stained according to a modified method of Dawson (1926).

A dose-related increase in maternal toxicity with increasing severeness of clinical signs (lateral and abdominal position, unsteady gait, salivation, piloerection, nasal discharge and pneumonia) was observed. A slight decrease in food consumption and body weight gain was observed at650,975,1300mg/kg day dose group. Uterine and placental weights, foetal weights and data were not influenced by administration of this material. No dead foetuses were observed. There was no indication of developmental toxicity. All foetal values were within the range of biological variation; any differences in malformations and retardations were statistically insignificant or without a dose-response relationship and mostly occurred in animals of the treated and untreated groups as well as the historical control group at comparable frequency. A single cheiloschisis and one anophthalmy in the 1300mg/kg bw dose group was considered coincidental and not biologically significant. Hencethe no observed adverse effect level (NOALE) was considered to be 1300mg/kg bw/day for  reproductive and developmental toxicity .When female rats were treated with test material orally during gestation day 6-15.

 

Based on the data available from different studies test chemical did not showedreproductive toxicityat dose concentration 1000mg/kg bw/day .When male and female rats were treated with test material orally.Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

 

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from authoritative database

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive toxicity study

Data available from different studies were reviewed to determine the reproductive toxicity of testchemical.The studies are as mentioned below:

Study 1.

The reproductive and developmental toxicity study of test material was performed in femaleCOBS CD rats. The test material was dissolved in corn oil and administered in dose concentration 0,200,1000mg/kg bw by oral gavage route from days 6-15 of gestation. 25 females /dose group were used in study. All the animals were observed forClinical signs, body weight and food consumoption.The dams were sacrificed and sectioned on gestation day 20. Intrauterine survival, foetal weight and external, skeletal and visceral anomalies were recorded. In dams clinical signs of intoxication were observed also reduced body weight was observed at 1000mg/kg bw dose group. Effects on the foetus were confined to an increase in numbers of litters with the skeletal variant 'malaligned sternebrae' which occurred at 200 mg/kg/day only and a slight decrease in foetal weight at 1000 mg/kg/day which was within the historical control range. As an increased incidence of 'malaligned sternebrae' was not observed at 1000 mg/kg/day the observation at 200 mg/kg/day was considered incidental. Hencethe no observed adverse effect level (NOAEL) was considered to be 1000mg/kg bw/day for reproductive and developmental toxicity .When female rats were treated with test material by orally during gestation days 6-15.

Study 2.

Combined Repeat Dose and Reproductive / Developmental Toxicity Screening Test of test material were performed on male and female Sprague Dawley rats. The test material was suspended in corn oil in dose concentration0 ,100, 300 and 1,000 mg/kg bw/day and administered via oral gavage route in dose volume 5ml/kg bw. Dose selected on the bases ofpreliminary test by the previously repeated 14-day oral administration. 13 male and 13 female each were placed in each group. Administration period For each males, 2 weeks before mating, and 42 days from the end of the mating period to the day before the necropsy for males, and for female 2 weeks before mating and a maximum of 2 weeks mating period (Until mating) and once daily for the entire gestation period and 3 days after nursing (delivery day = nursing 0 day) in mating females. Matings were made with the same sex of the same group living together within the same group for up to two weeks from the evening on 15th day of administration. Mating was confirmed every morning by examining the presence of sperm in the vaginal plug and vaginal smear and females confirmed to be mated were separated from male starting from that day as the 0th day of pregnancy and raised individually. All the animals were observed forClinical signs, body weight and food consumption.

No deaths in male and female were observed. In males, no abnormalities in general conditions were observed. In females, there was only one change from day 42 of administration (day of delivery) to the one in 100 mg / kg administration group (administration 42 days: piloerection, blood-like fluid discharge from the vaginal opening; administration 43 to 44 days: coat contamination) But it recovered on day 45 of administration. In other females, no abnormalities in the general condition were observed. In male, the body weight gain increased from 8 to 15 days in the 100 mg / kg administration group significantly (p <0.01) compared with the control group, However, in the group administered with 300 mg / kg or more, there was no significant difference between the control group and treated group. In females, there was no significant difference in body weight and body weight gain between control group and treated group before breeding, pregnancy period and nursing period. There was no significant difference between the control group and treated group for males and female at any time of feeding intake. There was no significant difference between the control group and treated group in the mating rate, conception rate, the number of days taken from the start of cohabitation to mating and the number of estrus periods recurred during that period.

An abnormality in labor condition (not treated with placenta) and abnormality in nursing condition (not collecting children, poor protrusion of nipple, poor protrusion of infant, child's fetus) was administered to one of 100 mg / kg administration group, The decrease in body surface temperature) was observed, and all children died on nursing day 2, but no abnormality was observed in other mother animals. For male In the 100 mg / kg administration group, the actual weight and the specific body weight value of the liver increased significantly (p <0.05) compared to the control group, but not the dose-dependent change. Atrophy of seminiferous tubules was found in 2 animals in the 1000 mg / kg administration group, one on both sides and one on one side, but both were localized and extremely mildly changed. There were no other abnormalities. Sperm granuloma was observed in one of the control group, and there was no abnormality. There were no abnormalities in the ovaries of the unexpanded and infertile cases. For female In the 100 mg / kg administration group, the actual weight of the kidney decreased significantly (p <0.05) compared to the control group, but not the dose-dependent change. For other organs in male and female, no significant difference was observed between the control group and treated group. Birth rate was 100% in all administration groups, and no significant difference was observed between gestation period between control group and treated group. No significant difference was observed between the control group and treated group for the number of luteinism, the number of implantation and the implantation rate of pregnant animals.There was no significant difference between the control group and treated group for the number of births, delivery rate, birth rate, fertility rate, and survival rate on 4th day of newborn. In addition, no significant difference was observed between the control group and treated group for sex ratio.There was no significant difference between the control group and thetreated grouponbody weight at 0 and 4 days of nursing. Births that showed morphological abnormalities were not observed in the external table observation on thebirthday, necropsy of the dead child, and necropsy on the 4th day of nursing. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 1000mg/kg bw .When male and female Sprague Dawley rats were treated with test material orally.

 Study 3.

The reproductive and developmental toxicity study of test material was performed on female wistar rats.The chemicals were freshly prepared for gavage administration every day in aqueous emulsions under rapid stirring in doubly-distilled water containing approximately 0.005% Cremophor EL". The test material in dose concentration0,130,650,975,1300mg/kg day was adminstered fromday 6 to day 15 post-coitum (pc). Two control groups, treated with either doubly-distilled water alone (1) or water plus approximately 0.005% Cremophor EL as emulsifier (2), were employed.one to four untreated females were mated with one untreated fertile male of the same breed. Mating took place overnight. If sperm was detected microscopically in the vaginal smears in the morning, the animals were considered to be fertilized. This day was designated as 'day 0' (beginning of the study) and the following day 'day 1' post-coitum (pc).Food consumption, body weights and clinical signs were recorded daily throughout the study. On day 20 pc all surviving females were killed and subjected to gross pathology.The foetuses were dissected from the uterus, weighed and further investigated for external, visceral and skeletal findings. Approximately one-half of the foetuses of all groups was fixed in Bouin's solution and examined according to the method of Barrow and Taylor (1969). For skeletal findings approximately one-half of the foetuses was fixed in ethyl alcohol and stained according to a modified method of Dawson (1926).

A dose-related increase in maternal toxicity with increasing severeness of clinical signs (lateral and abdominal position, unsteady gait, salivation, piloerection, nasal discharge and pneumonia) was observed. A slight decrease in food consumption and body weight gain was observed at650,975,1300mg/kg day dose group. Uterine and placental weights, foetal weights and data were not influenced by administration of this material. No dead foetuses were observed. There was no indication of developmental toxicity. All foetal values were within the range of biological variation; any differences in malformations and retardations were statistically insignificant or without a dose-response relationship and mostly occurred in animals of the treated and untreated groups as well as the historical control group at comparable frequency. A single cheiloschisis and one anophthalmy in the 1300mg/kg bw dose group was considered coincidental and not biologically significant. Hencethe no observed adverse effect level (NOALE) was considered to be 1300mg/kg bw/day for  reproductive and developmental toxicity .When female rats were treated with test material orally during gestation day 6-15.

 

Based on the data available from different studies test chemical did not showedreproductive toxicityat dose concentration 1000mg/kg bw/day .When male and female rats were treated with test material orally.Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

 

Effects on developmental toxicity

Description of key information

The oral administration of 100, 300 and 1000 mg/kg of docosanoic acid to SD rats produced no reproductive and developmental toxicity therefore, the No Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 1000mg/kg.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experemental data from various test chemicals

Justification for type of information:

Weight of evidence approach based on the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

Reproductive toxicity study of test material was performed on Sprague Dawley rats.

GLP compliance:

no

Limit test:

no

Species:

other: 2. rat 3. Rabbit

Strain:

other: 2. Sprague-Dawley(Crj: CD, SPF) 3. New Zealand White

Details on test animals or test system and environmental conditions:

2. Details on test animals and env. conditions
TEST ANIMALS
- Source: Charles River, Japan
- Age at study initiation: 8 weeks old
- Weight at study initiation: male 312.1 to 363.7 g, female 205.3 -230.8 g).
- Fasting period before study:
- Housing: individually housed in a wire mesh floor cage (Nihon Cage ), raised, and allowed to take solid feed
(CE - 2, Japan Clea )
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period:7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 1 ° C.
- Humidity (%):50 to 65%,
- Air changes (per hr): about 15 times / hour
- Photoperiod (hrs dark / hrs light): a lighting condition set at 12 hours of lighting (7 am to 7 pm)

3.TEST ANIMALS
- Source: Froxfield SPF Rabbits Ltd., UK
- Age at study initiation: 18-26 weeks on arrival
- Weight at study initiation: 3.29-4.98 kg at start of study
- Fasting period before study: no data
- Housing: individually in suspended stainless-steel cages (TR6)
- Diet (e.g. ad libitum): standard rabbit diet (Special Diets Services Ltd., UK), ad libitum
- Water (e.g. ad libitum): public supply, ad libitum
- Acclimation period: >=1 week
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18
- Humidity (%): 55
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12
IN-LIFE DATES: no data

Route of administration:

oral: gavage

Vehicle:

other: 2. corn oil 3. 1% Twin 80

Details on exposure:

2.Details on exposure
PREPARATION OF DOSING SOLUTIONS:
The test chemical was prepared by suspending it in corn oil and used as a test sample.
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil
- Concentration in vehicle: 0 ,100, 300 and 1,000 mg/kg bw/day
- Amount of vehicle (if gavage): 5ml/kg bw
- Lot/batch no. (if required): (Lot No. V 6 H 2050, Nacalai Tesque ),
- Purity: No data available

3.PREPARATION OF DOSING SOLUTIONS:
- test material weighed into glass container and heated to ~80 deg C until molten
- vehicle heated to 75 deg C
- test material and vehicle combined using coninuous magnetic stirring, 20% behenyl alcohol
- suspension cooled slowly to <60 deg C
- further cooled to 30 deg C
- slowly homogenized <=2 min
- cooled to room temperature
- 20% suspension prepared weekly
- 20% suspension provided top dose
- mid and low dose prepared on day of use by dilution with vehicle; 20% suspension magnetically
stirred prior to removal of aliquots for dilution; dilutions hand swirled prior to magnetic stirring
VEHICLE
- Justification for use and choice of vehicle (if other than water): not stated
- Concentration in vehicle: 20, 2 and 0.2%
- Amount of vehicle (if gavage): 10 ml/kg bw for vehicle control and top dose groups; 0.625 and
2.5 ml/kg bw for low and mid dose groups respectively
- Lot/batch no. (if required): no data
- Purity: 1%

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No Data Available

Details on mating procedure:

2. - M/F ratio per cage: 1/1
- Length of cohabitation: at the most 14days
- Further matings after two unsuccessful attempts: [no / yes (explain)] : no
- Verification of same strain and source of both sexes: [yes / no (explain)] : no
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy : until proof of pregnancy (formation of vaginal closing or sperm detection in vagina)

3.- Impregnation procedure: cohoused with males of establised fertility
- If cohoused:
- M/F ratio per cage: no data
- Length of cohabitation: no data
- After ... days of unsuccessful pairing replacement of first male by another male with proven
fertility: no data
- Further matings after two unsuccessful attempts: no data
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: not specified, but referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no data

Duration of treatment / exposure:

2.Males: 43 days; Females: from 14 days prior to mating to day 4 of lactation.
3. days 6-19 of gestation

Frequency of treatment:

Daily

Duration of test:

2. 43 days
3. females killed on day 29 of gestation

Remarks:

Doses / Concentrations: (2)
0, 100, 300, 1000 mg/kg/day (in corn oil)
Basis:nominal conc.

Remarks:

0, 125, 500 and 2000 Mg/kg Bw/day (3)

No. of animals per sex per dose:

2. Total ;104
0 (Vehicle): 13 male and 13 female
100 mg/kg/day: 13 male and 13 female
300 mg/kg/day: 13 male and 13 female
1000 mg/kg/day: 13 male and 13 female

3. 22 Females

Control animals:

yes, concurrent vehicle

Details on study design:

2. - Dose selection rationale: As a result of the preliminary test by the previously repeated 14-day oral administration, no signs of toxicity were observed even in the group administered with the critical dose of 1000 mg / kg prescribed in the OECD Chemical Test Method Guideline
- Rationale for animal assignment (if not random):
- Other: No Data Available

3.Sex: female
Duration of test: 28 days
- Dose selection rationale: based on previous range-finding study
- Rationale for animal assignment (if not random): randomly allocated to the four treatment
groups in order of mating "to evenly distribute the mated females among the groups"
- Other:
- approximately 2 weeks prior to arrival of females at testing facility, oestrus synchronised by
supplier by intravenous injection of 25 IU luteinizing hormone
- following insemination, females injected intravenously with 25 IU luteinizing hormone to ensure
successful ovulation
- examined on day 6 of gestation, prior to dosing, to determine suitability for use in study

Maternal examinations:

2. Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS:
Time schedule: Daily

BODY WEIGHT: Yes
Time schedule for examinations: For all males, measurements were made on 1, 8, and 15 days of administration 1 (administration start date), 8, 15, 22, 29, 36, 42 days and postmenopausal days and females for all cases,

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): yes For all males, measurements were made on 1, 8, and 15 days of administration (administration start date), 8, 15,22, 29, 36, 42 days and postmenopausal days and females for all cases,
Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
Time schedule for examinations:

3. CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations included: evidence of reaction to treatment or moribund condition
DETAILED CLINICAL OBSERVATIONS: no data
BODY WEIGHT: Yes
- Time schedule for examinations: daily
FOOD CONSUMPTION: Yes
- Time schedule for examinations: days 1-5, days 6-12, days 13-19, days 20-23, days 24-28
WATER CONSUMPTION: Yes
- Time schedule for examinations: daily
POST-MORTEM EXAMINATIONS: yes, macroscopic examination
- Sacrifice on gestation day 29
- Organs examined in addition to uterine contents and ovaries: no data

Ovaries and uterine content:

2.The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: No Data Available

3.The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:
- Number of viable young: males, females and total
- Distribution of foetusus in each uterine horn
- Uterus of any female presumed non-pregnant stained and examined for implantation sites

Fetal examinations:

2. - External examinations: Yes: all per litter
- Soft tissue examinations: Yes:
- Skeletal examinations: Yes:
- Head examinations: No data

3.- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: all per litter - cervical, thoracic and abdominal cavities dissected
and contents examined microscopically
- Skeletal examinations: Yes: all per litter
- Head examinations: Yes: one third per litter
- Other: all per litter
- foetal body weight
- position of foetus in uterus
- placental weight

Statistics:

2. Dunnett’s or Scheffe’s test for continuous data, Chi square test for copulated index and fertility index, and Mann-Whitney U test or Fisher’s test for histopathological examination data.

3. One-way analysis of variance, t-tests - body weight, body weight change, food and water
consumption; Dunnett's or Behren's-Fisher's tests - organ weights; nested analysis of variance,
weighted t-tests - foetal and placental weights

Indices:

2. Copulation index, fertility index, implantation index, gestation index, delivery index, birth index sex ratio, viability index
3. No data

Historical control data:

No data available

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

2. In males, no abnormalities in general conditions were observed. In females, there was only one change from day 42 of administration (day of delivery) to the one in 100 mg / kg administration group (administration 42 days: piloerection, blood-like fluid discharge from the vaginal opening; administration 43 to 44 days: coat contamination) But it recovered on day 45 of administration. In other females, no abnormalities in the general condition were observed.

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

2. In male, the body weight gain increased from 8 to 15 days in the 100 mg / kg administration group significantly (p <0.01) compared with the control group, However, in the group administered with 300 mg / kg or more, there was no significant difference between the control group and treated group. In females, there was no significant difference in body weight and body weight gain between control group and treated group before breeding, pregnancy period and nursing period.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

2. There was no significant difference between the control group and treated group for males and female at any time of feeding intake.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

2. For female In the 100 mg / kg administration group, the actual weight of the kidney decreased significantly (p <0.05) compared to the control group, but not the dose-dependent change. For other organs in male and female, no significant difference was observed between the control group and treated group.

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

2. There were no abnormalities in the ovaries of the unexpanded and infertile cases.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Details on results:

No Data Available

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Description (incidence and severity):

2. No significant difference was observed between the control group and treated group for the number of luteinism, the number of implantation and the implantation rate of pregnant animals.

Total litter losses by resorption:

not specified

Early or late resorptions:

not specified

Dead fetuses:

no effects observed

Description (incidence and severity):

2. Birth rate was 100% in all administration groups.

Changes in pregnancy duration:

no effects observed

Description (incidence and severity):

2. No significant difference was observed between gestation period between control group and treated group.
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.DescriptionIncidenceAndSeverityEffectsOnPregnancyDuration): No significant difference was observed between gestation period between control group and treated group.

Changes in number of pregnant:

no effects observed

Other effects:

not specified

Details on maternal toxic effects:

2. Maternal toxic effects:no effects. Remark: No abnormalities were found.

Details on maternal toxic effects:
No abnormalities were found in all reproductive parameters (fertility index, number of Implantations and implantation index) in each dose group. No statistically significant difference from controls in the case of Body weight,Food/water consumption and Reproductive responses.

3.Maternal toxic effects:no effects
Details on maternal toxic effects:
no effects other than pale faeces in animals of the top dose group

Key result

Dose descriptor:

NOAEL

Remarks:

2.

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

body weight and weight gain

changes in number of pregnant

changes in pregnancy duration

clinical biochemistry

clinical signs

dead fetuses

early or late resorptions

effects on pregnancy duration

food consumption and compound intake

gross pathology

haematology

histopathology: non-neoplastic

mortality

number of abortions

organ weights and organ / body weight ratios

pre and post implantation loss

Remarks on result:

other: overall no effects on reproductive performance

Dose descriptor:

NOAEL

Remarks:

3

Effect level:

2 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: Maternal toxicity

Abnormalities:

not specified

Localisation:

not specified

Fetal body weight changes:

no effects observed

Description (incidence and severity):

2. There was no significant difference between the control group and the treated group on body weight at 0 and 4 days of nursing.

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified

Reduction in number of live offspring:

no effects observed

Description (incidence and severity):

2. There was no significant difference between the control group and treated group for the number of births, delivery rate, birth rate and survival rate on 4th day of newborn.

Changes in sex ratio:

no effects observed

Description (incidence and severity):

2. no significant difference was observed between the control group and treated group for sex ratio.

Changes in litter size and weights:

not specified

Changes in postnatal survival:

no effects observed

Description (incidence and severity):

2. There was no significant difference between the control group and treated group for survival rate on 4th day of newborn.

External malformations:

no effects observed

Description (incidence and severity):

2. No morphological abnormalities in pups were observed in any of the treated groups.

Skeletal malformations:

not specified

Visceral malformations:

not specified

Other effects:

not specified

Details on embryotoxic / teratogenic effects:

2. Embryotoxic / teratogenic effects:no effects. Remark: No abnormalities were found in all pups.

Details on embryotoxic / teratogenic effects:
The compound did not demonstrate any adverse effects on the sex ratio, body weights or viability of pups. Also, no morphological abnormalities in pups were observed in any of the treated groups.

3.Embryotoxic / teratogenic effects:no effects
Details on embryotoxic / teratogenic effects:
no effects

Dose descriptor:

NOAEL

Remarks:

2

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

reduction in number of live offspring

changes in sex ratio

changes in litter size and weights

changes in postnatal survival

external malformations

Remarks on result:

other: overall no developmental toxic effects observed

Dose descriptor:

NOAEL

Effect level:

2 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: Teratogenecity

Abnormalities:

not specified

Localisation:

other: not specified

Developmental effects observed:

not specified

Treatment related:

not specified

Conclusions:

The oral administration of 100, 300 and 1000 mg/kg of docosanoic acid to SD rats produced no reproductive and developmental toxicity therefore, the No Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 1000mg/kg.

Executive summary:

Developmental toxicity study was performed on Rats and Rabbits to determine the toxic nature of the test substance. The toxicity of the test chemical is sufficed by applying Weight of evidence approach. The studies are summarized below as follows:

2. A combined repeated dose toxicity and reproductive/developmental screening test of Docosanoic acid was performed on Sprague-Dawleyrats. Groups of 13 rats/sex/dose were orally administered (gavage) 0, 100, 300 and 1000 mg of docosanoic acid per kg. The test chemical was suspended in corn oil and used in volume of 5 ml/kg bw. The doses were selected on the bases of a preliminary, 14-day repeated dose toxicity study. In male rats, the administration period was two weeks prior to mating, 2 weeks of mating and 2 weeks after the completion of the mating period (42 days). In females, in addition to maximum four weeks pre-mating and mating period, they were exposed through pregnancy until day 3 of post-delivery (PDN 3). Mating was confirmed by the presence of sperm in the vaginal plug or in the vaginal smear. Females were allowed to deliver spontaneously and the conditions of parturition such as labour difficulty or delay and nursing behaviour were observed. No deaths or clinical signs of toxicology were observed in any male and female animals. The body weight and food consumption of treated animals, both males and females, were comparable with controls. At day 43 male animals were necropsied, most of the organs were weighed including reproductive organs (testis and epididymis) then tissues were fixed, and histopathological examination were performed. Before necropsy blood was collected for clinical biochemistry and haematology. The mean red blood cell haemoglobin concentration (MCHC) decreased significantly compared to the control group in the 300 and 1000 mg/kg dose groups, but no further significant difference was observed. The alkaline phosphates activity significantly decreased in all docosanoic acid administration groups and the glucose level was significantly lower in 1000 mg/kg group than in the control. However, alterations of blood parameters were not accompanied by any changes found in other examination including pathological examination, therefore they are considered to be accidental changes and have no toxicological significance. At necropsy there were no obvious changes that could be attributed to administration of docosanoic acid. No significant differences in organ weights were found between the control group and dosed animals. No treatment-related abnormalities were observed in adrenal gland, testis and epididymis in any treated males. Atrophy of the seminiferous tubule was observed in two animals at 1000 mg/kg, but the lesions were localized and very mild. There were no other abnormalities in male reproductive organs. Female animals (both mated and infertile) were also necropsied and organ weights were measured for heart, liver, kidney, and thymus. The ovaries and uterus were removed and the number of corpora lutea, the number of implantation and the implantation rate were calculated. Organs were fixed and tissue samples were prepared for further histopathological analysis. No obvious changes attributable to administration of docosanoic acid were seen in autopsy findings and organ weights. There were no significant differences in the number of corpora lutea, the number of implantations and the implantation rate of the pregnant animals between the control group and each administration group of docosanoic acid. The copulation and fertility indexes of each dose were comparable to controls. There were no significant differences between the control group and each dose of docosanoic acid in the mating rate, conception rate, the number of days required from the start of cohabitation to mating, and the number of oestrus returned during that period. The birth rate was 100% in all treatment groups. The number of births, delivery rate, live birth rate, and 4-day survival rate of newborns were not significantly different between the control and dosed groups. No significant difference in sex ratio of pups between the control and treated groups were seen. Similarly, there were no significant differences in body weight on day 0 and 4 between the control and docosanoic acid groups. No offspring with morphological abnormalities were observed at PND 0 and 4 following external surface observation and autopsy. In summary, no abnormality was found in fertility, reproductive function and parturition of parental animals and the prenatal and early postnatal exposure to the test chemical did not produced adverse effects on neonatal survival and growth, when male and female SD rats were orally administered of 100, 300 and 1000 mg/kg of docosanoic acid. Hence, the NOAEL for reproductive and developmental toxicity is considered as 1000 mg/kg under the certain experimental conditions.

3.

A developmental toxicity study was performed on Rabbits according to OECD guideline 414. 22 New Zealand white Female rabbits were treated with test chemical at a concentration of 0, 125, 500 and 2000 mg/kg bw/day for a duration of 28 days. The vehicle used was 1% twin 80 and the test chemical was administerd orally via gavage daily. Females were killed at day 29 of gestation period. The test chemical had no adverse effects on the general condition, body weight gains, or food and water consumption of the dams, and was without consequence to various reproductive parameters, including number of corpora lutea, pre- and postimplantation sites, early and late resorptions, and viable fetuses. In conclusion, an embryonic development study in rabbits administered a maximum dose of 2000 mg/kg body weight provide no evidence to suggest that the test chemical would be teratogenic or embryotoxic.

A developmental toxicity study was performed on Rabbits according to OECD guideline 414. 22 New Zealand white Female rabbits were treated with test chemical at a concentration of 0, 125, 500 and 2000 mg/kg bw/day for a duration of 28 days. The vehicle used was 1% twin 80 and the test chemical was administerd orally via gavage daily. Females were killed at day 29 of gestation period. The test chemical had no adverse effects on the general condition, body weight gains, or food and water consumption of the dams, and was without consequence to various reproductive parameters, including number of corpora lutea, pre- and postimplantation sites, early and late resorptions, and viable fetuses. In conclusion, an embryonic development study in rabbits administered a maximum dose of 2000 mg/kg body weight provide no evidence to suggest that the test chemical would be teratogenic or embryotoxic.

Hence, the NOAEL for reproductive and developmental toxicity is considered as 1000 mg/kg under the certain experimental conditions.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

K2

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

Developmental toxicity study was performed on Rats and Rabbits to determine the toxic nature of the test substance. The toxicity of the test chemical is sufficed by applying Weight of evidence approach. The studies are summarized below as follows:

2. A combined repeated dose toxicity and reproductive/developmental screening test of Docosanoic acid was performed on Sprague-Dawleyrats. Groups of 13 rats/sex/dose were orally administered (gavage) 0, 100, 300 and 1000 mg of docosanoic acid per kg. The test chemical was suspended in corn oil and used in volume of 5 ml/kg bw. The doses were selected on the bases of a preliminary, 14-day repeated dose toxicity study. In male rats, the administration period was two weeks prior to mating, 2 weeks of mating and 2 weeks after the completion of the mating period (42 days). In females, in addition to maximum four weeks pre-mating and mating period, they were exposed through pregnancy until day 3 of post-delivery (PDN 3). Mating was confirmed by the presence of sperm in the vaginal plug or in the vaginal smear. Females were allowed to deliver spontaneously and the conditions of parturition such as labour difficulty or delay and nursing behaviour were observed. No deaths or clinical signs of toxicology were observed in any male and female animals. The body weight and food consumption of treated animals, both males and females, were comparable with controls. At day 43 male animals were necropsied, most of the organs were weighed including reproductive organs (testis and epididymis) then tissues were fixed, and histopathological examination were performed. Before necropsy blood was collected for clinical biochemistry and haematology. The mean red blood cell haemoglobin concentration (MCHC) decreased significantly compared to the control group in the 300 and 1000 mg/kg dose groups, but no further significant difference was observed. The alkaline phosphates activity significantly decreased in all docosanoic acid administration groups and the glucose level was significantly lower in 1000 mg/kg group than in the control. However, alterations of blood parameters were not accompanied by any changes found in other examination including pathological examination, therefore they are considered to be accidental changes and have no toxicological significance. At necropsy there were no obvious changes that could be attributed to administration of docosanoic acid. No significant differences in organ weights were found between the control group and dosed animals. No treatment-related abnormalities were observed in adrenal gland, testis and epididymis in any treated males. Atrophy of the seminiferous tubule was observed in two animals at 1000 mg/kg, but the lesions were localized and very mild. There were no other abnormalities in male reproductive organs. Female animals (both mated and infertile) were also necropsied and organ weights were measured for heart, liver, kidney, and thymus. The ovaries and uterus were removed and the number of corpora lutea, the number of implantation and the implantation rate were calculated. Organs were fixed and tissue samples were prepared for further histopathological analysis. No obvious changes attributable to administration of docosanoic acid were seen in autopsy findings and organ weights. There were no significant differences in the number of corpora lutea, the number of implantations and the implantation rate of the pregnant animals between the control group and each administration group of docosanoic acid. The copulation and fertility indexes of each dose were comparable to controls. There were no significant differences between the control group and each dose of docosanoic acid in the mating rate, conception rate, the number of days required from the start of cohabitation to mating, and the number of oestrus returned during that period. The birth rate was 100% in all treatment groups. The number of births, delivery rate, live birth rate, and 4-day survival rate of newborns were not significantly different between the control and dosed groups. No significant difference in sex ratio of pups between the control and treated groups were seen. Similarly, there were no significant differences in body weight on day 0 and 4 between the control and docosanoic acid groups. No offspring with morphological abnormalities were observed at PND 0 and 4 following external surface observation and autopsy. In summary, no abnormality was found in fertility, reproductive function and parturition of parental animals and the prenatal and early postnatal exposure to the test chemical did not produced adverse effects on neonatal survival and growth, when male and female SD rats were orally administered of 100, 300 and 1000 mg/kg of docosanoic acid. Hence, the NOAEL for reproductive and developmental toxicity is considered as 1000 mg/kg under the certain experimental conditions.

3.

A developmental toxicity study was performed on Rabbits according to OECD guideline 414. 22 New Zealand white Female rabbits were treated with test chemical at a concentration of 0, 125, 500 and 2000 mg/kg bw/day for a duration of 28 days. The vehicle used was 1% twin 80 and the test chemical was administerd orally via gavage daily. Females were killed at day 29 of gestation period. The test chemical had no adverse effects on the general condition, body weight gains, or food and water consumption of the dams, and was without consequence to various reproductive parameters, including number of corpora lutea, pre- and postimplantation sites, early and late resorptions, and viable fetuses. In conclusion, an embryonic development study in rabbits administered a maximum dose of 2000 mg/kg body weight provide no evidence to suggest that the test chemical would be teratogenic or embryotoxic.

A developmental toxicity study was performed on Rabbits according to OECD guideline 414. 22 New Zealand white Female rabbits were treated with test chemical at a concentration of 0, 125, 500 and 2000 mg/kg bw/day for a duration of 28 days. The vehicle used was 1% twin 80 and the test chemical was administerd orally via gavage daily. Females were killed at day 29 of gestation period. The test chemical had no adverse effects on the general condition, body weight gains, or food and water consumption of the dams, and was without consequence to various reproductive parameters, including number of corpora lutea, pre- and postimplantation sites, early and late resorptions, and viable fetuses. In conclusion, an embryonic development study in rabbits administered a maximum dose of 2000 mg/kg body weight provide no evidence to suggest that the test chemical would be teratogenic or embryotoxic.

Hence, the NOAEL for reproductive and developmental toxicity is considered as 1000 mg/kg under the certain experimental conditions.

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive and Developmental toxicant.