Registration Dossier

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

There are reliable two screening studies for reproducitve and developmental toxicity performed with a constituent of the registration substance and a structural analogue avalaible:

1. Combined repeated dose toxicity study with the reproduction/developmental screening test according to OECD 422 performed with tetradecyl oleate (CAS No 22393-85-7), constituent of the registration substance

NOAEL(systemic, reproduction, developmental): > 1000 mg/kg bw/d (male/female)

2. Combined repeated dose toxicity study with the reproduction/developmental screening test according to OECD 422 performed with docosyl docosanoate (CAS No 17671-27-1), strucutral analogue of the substance

NOAEL(systemic, reproduction, developmental): > 1000 mg/kg bw/d

Link to relevant study records

Referenceopen allclose all

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
06 Jun - 04 Aug 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
adopted in 22 Mar 1996
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl:CD(SD)BR
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Italia S.p.A., Calco (Lecco), Italy
- Age at study initiation: 6 to 7 weeks
- Weight at study initiation: 191 to 204 g (males) and 172 to 179 g (females)
- Housing: From arrival to pairing: animals were housed 5 of one sex to a cage, in polysulphone solid bottomed cages measuring 59.5x38x20 cm (Techniplast Gazzada S.a.r.l., Buguggiate, Varese, Italy). Nesting material was provided inside suitable bedding bags and changed at least twice a week.
During mating: animals were housed one male to one female in clear polysulphone cages measuring approximately 43x27x18 cm with a stainless steel mesh lid and floor (Techniplast Gazzada S.a.r.l., Buguggiate, Varese, Italy). Each cage tray held absorbent material which was inspected and changed daily. After mating, the males were re-caged as they were before mating; the females were transferred to individual solid bottomed cages (Techniplast Gazzada S.a.r.l., Buguggiate, Varese, Italy) for the gestation period and parturition.
- Diet: laboratory rodent diet, 4 RF 21 (Mucedola S.r.l., Settimo Milanese (MI), Italy), ad libitum
- Water: drinking water, ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 15
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: gavage
Vehicle:
other: 0.5% aqueous carboxymethylcellulose (0.5% CMC)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The test substance was suspended in the vehicle. Formulations were prepared daily.

VEHICLE
- Concentration in vehicle: 10, 30 and 100 mg/mL for dose levels of 100, 300 and 1000 mg/kg bw/day, respectively
- Amount of vehicle: 10 mL/kg bw
Details on mating procedure:
- M/F ratio per cage: 1 male to 1 female (monogamous).
- Length of cohabitation: The female was placed with the same male until pregnancy had occurred or 2 weeks had elapsed.
- Proof of pregnancy: Vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy.
- After 2 weeks of unsuccessful pairing replacement of first male by another male with proven fertility.
- After successful mating each pregnant female was caged singly.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Concentration, homogeneity and stability of the test substance in the vehicle were verified by gas chromatopgraphy with a flame ionisation detection (FID). Concentration verification was conducted on a weekly basis.
Duration of treatment / exposure:
Males: The daily administration of the test item was started two weeks before mating and lasted until test day 28 to 29, which was one day before sacrifice.
Females: The daily administration of the test item was started two weeks before mating and continued until day 3 post-partum.
Maximum: 54 days of treatment.
Frequency of treatment:
once daily; 7 days/week
Details on study schedule:
not applicable for OECD 422 study
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on a 14-day range-finding study (Rossiello, 2013. RTC Study No.: 93730EXT)
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once daily during the study, each animal was observed and any clinical signs recorded.


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once before commencement of treatment and at least once a week thereafter, each animal was given a detailed clinical examination. Each animal was removed from the home cage and observed in an open arena. The tests included observation of changes in gait and posture, reactivity to handling, presence of clonic or tonic movements, stereotypes or bizarre behaviour and effects on the autonomic nervous system (e.g. lachrymation, piloerection, pupil size, unusual respiratory pattern).

BODY WEIGHT: Yes
- Time schedule for examinations: females: weekly from allocation to positive identification of mating and on gestation Days 0, 7, 14 and 20. Dams were also weighed on Days 1 and 4 post partum.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: The weight of food consumed by each cage of males and females was recorded weekly during the pre-mating period starting from allocation. Individual food consumption for the females was measured on gestation Days 7, 14 and 20 starting from Day 0 post coitum and on Day 4 post partum starting from Day 1 post partum.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

For further observations and examinations (water intake, haematology, clinical chemistry, neurobehaviour), see "Repeated dose toxicity: oral" (chapter 7.5.1)

OTHER:
Reproduction paramters: number of pregnant females, pre-coital time, gestation length
Oestrous cyclicity (parental animals):
Vaginal smears were taken daily in the morning starting two weeks before pairing until a positive identification of copulation was made. The vaginal smear data were examined to determine the following: anomalies of the oestrous cycle and pre-coital interval (i.e., the number of nights paired prior to the detection of mating).
Sperm parameters (parental animals):
Parameters examined in P male parental generations:
testis weight, epididymis weight, and qualitative sperm staging.
In addition, the testes and epididymides were cut at 2-3 micrometer thickness and stained with Periodic Acid Schiff (PAS). The morphological evaluation of the seminiferous epithelium (staging of spermatogenic cycle) was performed. A detailed qualitative evaluation of testes was performed on 5 randomly selected control and high dose males. The evaluation took into account the tubular stages of the spermatogenic cycle, in order to identify treatment-related effects such as: missing germ cell layers or types, retained spermatids, multinucleated or apoptotic germ cells and sloughing of spermatogenic cells into the lumen.
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: litter weight, number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies

GROSS EXAMINATION OF DEAD PUPS:
Yes, for external and internal abnormalities
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals [males were sacrificed on day 29 or 30]
- Maternal animals: All surviving animals [females were sacrifices on day 4 post-partum or shorty thereafter]

GROSS PATHOLOGY: Yes
-Organ weights: adrenal glands, brain (cerebrum, cerebellum, medulla/pons), epididymides, heart, kidneys, liver, ovaries with oviducts, parathyroid glands, prostate gland, seminal vesicles with coagulating glands, spleen, testes, thymus (where present), thyroid and uterus-cervix,
-Fixation: adrenal glands, bone marrow (from sternum), brain (cerebrum, cerebellum, medulla/pons), caecum, clitoral gland, colon, duodenum, epididymides, heart, ileum (including Peyer’s patches), jejunum, kidneys, liver, lungs (including mainstem bronchi), lymph nodes (mesenteric and cervical), ovaries with oviducts, parathyroid glands, pituitary gland, penis, preputial gland, prostate gland, rectum, sciatic nerve, seminal vesicles with coagulating glands, spinal column, spinal cord (cervical, thoracic, lumber), spleen, stomach, testes, thymus (where present), thyroid, trachea, urinary bladder, uterus-cervix and vagina.

HISTOPATHOLOGY: Yes, all organs that were included for fixation (5/sex of control and high dose group)
Postmortem examinations (offspring):
SACRIFICE
- The surviving F1 offspring were sacrificed at 4 days of age.


GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations].
Dead pups and pups sacrificed at day 4 post-partum, or shortly thereafter, were carefully examined externally for gross abnormalities.

HISTOPATHOLOGY / ORGAN WEIGTHS
not performed
Statistics:
Standard deviations were calculated as appropriate. For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data. The non-parametric Kruskal-Wallis analysis of variance was used for the other parameters. Intergroup differences between the control and treated groups were assessed by the nonparametric version of the Williams test.
The criterion for statistical significance was p<0.05
Reproductive indices:
Male Copulatory Index (%) = No. of animals mated/No. of animals paired x 100
Male Fertility Index (%) = No. of males which induced pregnancy/ No. of males paired x 100
Female Copulatory Index (%) = No. of animals mated/No. of animals paired x 100
Female Fertility Index (%) = No. of pregnant females/No. of females paired x 100
Males and females:
Precoital interval = Mean number of days between pairing and mating
Offspring viability indices:
Pre-implantation loss [%] = (No. of corpora lutea - No. of implantations/ No. of corpora lutea) x 100
Pre-birth loss [%] = (No. of visible implantations - total litter size at birth/ No. of visible implantations
) x 100
Pup loss at birth [%] = (Total litter size - live litter size/ Total litter size) x 100
Cumulative pup loss on Day 4 post-partum [%] = (Total litter size at birth - live litter size at Day 4/ Total litter size at birth) x 100
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)

No relevant clinical signs or mortality were observed in males and females throughout the study.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)

No difference of toxicological significance were seen in body weight or body weight gain. No intergroup differences were seen in food consumption.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)

No relevant difference in oestrous cycle was observed in treated females when compared to controls.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)

Seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages; regular layering in the germinal epithelium was noted.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)

The number of corpora lutea, implantations, total litter size, pre-implantation loss and pre-birth loss did not differ significantly between groups. Gestation length was also comparable between groups. No differences were observed in the pre-coital interval, copulatory and fertility indices between control and treated groups.

ORGAN WEIGHTS (PARENTAL ANIMALS)

No relevant differences in terminal body weight or organ weights were seen between the controls and treated animals of both sexes

GROSS PATHOLOGY (PARENTAL ANIMALS)

No remarkable changes were noted at post mortem examination in treated animals when compared with controls.

HISTOPATHOLOGY (PARENTAL ANIMALS)

No treatment-related changes were observed. The lesions reported in control and treated animals were considered to be an expression of spontaneous and/or incidental pathology, commonly seen in this species and age under the experimental conditions.

Key result
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects obseved in the study
Key result
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed in the study
Key result
Critical effects observed:
no
Clinical signs:
no effects observed
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
All pregnant females gave birth to live pups with the exception of one high dose female which had a total litter loss on day 3 post-partum. This female and two control females had unilateral implantation but was not treatment related.
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:
not examined
Histopathological findings:
not examined
VIABILITY (OFFSPRING)

All pregnant females gave birth to live pups with the exception of one high dose female which had a total litter loss on day 3 post-partum. This female and two control females had unilateral implantation. This finding was considered incidental since it was observed also in two control females.

CLINICAL SIGNS (OFFSPRING)

Clinical signs of pups such as pallor, cold to touch, small and/or bruise muzzle, were observed in control, mid- and high dose groups. No toxicological relevance was attributed to these signs since they were seen in treated as well as in control groups.

BODY WEIGHT (OFFSPRING)

Litter data including mean litter and pup weights were comparable between groups. Sex ratio of pups showed a slight increased number of males in high dose group respect to control. No toxicological relevance was attribute to the statistical significant increase observed on Day 4.

GROSS PATHOLOGY (OFFSPRING)

Decedent pups were generally autolysed. No signs were seen in pups sacrificed on Day 4 post partum.
Key result
Dose descriptor:
NOAEL
Remarks:
developmental
Generation:
F1
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed in the study
Key result
Critical effects observed:
no
Key result
Reproductive effects observed:
no

Table 1: Fate of females: Group incidence

 

Treatment (mg/kg/bw/d)

0

100

300

1000

Initial group site

10

10

10

10

Unilateral Implantation

2

0

0

1

Total litter loss

0

0

0

1

With live pups on day 4 post-partum

10

10

10

9

Table 2: Implantation, pre-implantation loss data, pre-birth loss data and gestation length of females – Group mean data

Treatment (mg/kg/bw/d)

 

Corpora Lutea

Implantations

Total Litter size at birth

Pre-implantation loss %

Pre-birth loss %

Gestation length (days)

0

Mean

18.40

18.10

15.60

1.44

12.70

22.10

 

SD

3.37

3.18

5.27

3.17

25.82

0.32

 

n

10

10

10

10

10

10

 

100

Mean

16.30

15.80

14.20

3.26

10.09

22.10

 

SD

3.23

3.61

3.49

8.44

8.48

0.32

 

n

10

10

10

10

10

10

 

300

Mean

16.90

16.90

15.60

0.00

7.65

22.0

 

SD

1.97

1.97

2.17

0.00

7.35

0.00

 

n

10

10

10

10

10

10

 

1000

Mean

18.10

17.40

16.40

6.16

8.21

22.10

 

SD

5.17

5.32

5.44

12.73

9.93

0.32

 

n

10

10

10

10

10

10

Table 3: Litter data at birth, on day 1 and on day 4 post-partum of pregnant females – Group mean data

Treatment (mg/kg/bw/d)

 

At birth

On day 1 post-partum

On day 4 post-partum

Total litter size

Live litter size

Pup loss (%)

Litter weight (g)

Mean pup weight (g)

Live litter size

Cumulative loss (%)

Litter weight (g)

Mean pup weight (g)

0

Mean

15.60

15.40

1.13

101.62

7.00

14.40

6.37

132.38

9.61

SD

5.27

5.19

2.40

30.0

1.09

4.60

7.58

36.25

1.58

n

10

10

10

10

10

10

10

10

10

100

Mean

14.20

14.20

0.00

96.87

7.01

14.10

0.56

145.70

10.51

SD

3.49

3.49

0.00

19.08

0.81

3.38

1.77

29.27

1.08

n

10

10

10

10

10

10

10

10

10

300

Mean

15.60

15.60

0.00

106.70

6.94

14.80

5.51

143.13

9.76

SD

2.17

2.17

0.00

13.11

0.44

2.66

5.60

22.54

0.97

n

10

10

10

10

10

10

10

10

10

1000

Mean

16.40

16.20

5.50

115.50

7.10

15.60

13.30

163.63

9.46

SD

5.44

5.67

15.71

41.59

0.55

5.82

30.67

22.38

0.74

n

10

10

10

10

10

10

10

9

9

Table 4: Sex ratio of pups – Group mean data

Treatment (mg/kg/bw/d)

 

At birth

On day 4 post-partum

Males

Females

Total

% Males

Males

Females

Total

% Males

0

Mean

6.90

8.70

15.60

49.67

6.40

8.00

14.40

49.66

SD

2.02

4.03

5.27

19.96

1.71

3.59

4.60

19.93

n

10

10

10

10

10

10

10

10

100

Mean

6.90

7.30

14.20

50.50

6.80

7.30

14.10

50.14

SD

1.79

2.95

3.49

15.38

1.81

2.95

3.38

15.72

n

10

10

10

10

10

10

10

10

300

Mean

6.20

9.40

15.60

39.23

5.90

8.90

14.80

39.36

SD

2.20

1.65

2.17

11.28

2.08

1.66

2.66

10.29

n

10

10

10

10

10

10

10

10

1000

Mean

9.10

7.30

16.40

55.58

9.56*

7.78

17.33

55.66

SD

3.57

3.62

5.44

12.47

2.24

2.86

2.06

12.97

n

10

10

10

10

9

9

9

9

*= mean value of group is significantly different from control 

Conclusions:
Based on the result of this OECd 422 study performed with tetradecyl oelate (CAS No 22393-85-7), a constituent of the registration substance, the NOAEL was considered to be > 1000 mg/kg bw/d. No tretement-related effects were observed.
Executive summary:

Tetradecyl oelate (CAS No 22393-85-7), a constituent of the registration substance, was evaluated for its reprotoxic and developmental toxic potential in a combined subacute oral toxicity and reproduction/developmental toxicity screening study according to OECD 422.

Groups of 10 rats per sex and dose level were dosed with 0, 100, 300 and 1000 mg/kg bw/d of the test substance.

Since no effects were observed up to the highest dose level the NOAEL was derived at 1000 mg/kg bw/d.

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
18 Dec 2012 - 29 Jan 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
Please refer to cread across justification justification setcion 13.2

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Please refer to cread across justification justification setcion 13.2

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Please refer to cread across justification justification setcion 13.2

3. ANALOGUE APPROACH JUSTIFICATION
Please refer to cread across justification justification setcion 13.2

4. DATA MATRIX
Please refer to cread across justification justification setcion 13.2
Reason / purpose for cross-reference:
read-across source
Clinical signs:
no effects observed
Description (incidence and severity):
No mortality and no relevant clinical signs were observed in males and females. One male and female from the control group and three males and one female at 100 mg/kg showed occasionally up of the product. In addition, male No. 28 at 300 mg/kg had missing upper incisors in week 4 of treatment. No clinical signs were observed in females at 300 or 1000 mg/kg, while in males no clinical signs were oberved at 1000 mg/kg.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
not examined
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No mortality and no relevant clinical signs were observed. One female from the control group, one female and three males at 100 mg/kg bw showed occasionally up of the product. In addition, one male at 300 mg/kg bw had missing upper incisors in week 4 of treatment. No clinical sings were observed at 1000 mg/kg bw in either males or females.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
No test-item-related differences from the control group were recorded for body weight. Some significant differences were observed in group 3 but these were not considered of toxicological relevance.
Food consumption was similar in all groups.

REPRODUCTIVE FUNCTION:

SPERM MEASURES (PARENTAL ANIMALS)
All findings recorded were within the range of normal background lesions which may be recorded in animals of this strain and age. There were only single cases with isolated tubules that showed minor changes. They consisted of single sperm resorption in stage VII. In epididymides, there was only one animal in group 1 with unilateral monounclear cell foci of a minimal serverity.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Mating Performance: Median precoital time and mean precoital time (approx. 3 days) were similar in animals treated with the test item at 100 and 1000 mg/kg bw with respect to the control group. However, mean and median precoital time was longer at 300 mg/kg bw (5.7 and 4 days, respectively). In addition, female no. 68 from this group did not mate with the first male (no. 28) after a pairing period of 14 days and was mated a second time with male no. 23. This female showed an anestrus cycle of 11 days. Moreover, some animals from all groups took more than 2-4 days to mate. Females nos. 43 (control group), 53 (100 mg/kg), 61 and 70 (300 mg/kg) and 73 (1000 mg/kg) took 12, 12, 13, 14 and 13 days, respectively, to mate and showed an anestrus cycle.

Fertility: No treatment-related differences were recorded in the percentage of mating or the gestation index in females at the three doses with respect to the control group. Although fertility index and conception rate was lower in test-item-treated groups (100% vs. 90%), it was considered to be within the range of normal background findings which may be seen in rats.
Females no. 53 at 100 mg/kg bw, no. 70 at 300 mg/kg bw and no. 77 at 1000 mg/kg bw, which mated with males nos. 13, 30 and 37, respectively, were not pregnant despite the presence of sperm in the vaginal smear and vaginal plug in the last two females. Duration of the mating period of those couples was 12, 14 and 4 days, respectively. Females no. 69 at 300 mg/kg bw and no. 74 at 1000 mg/kg bw, which mated with males nos. 29 and 34, did not show signs of pregnancy and were mated again to ensure eight pregnant females per group. These animals were identified as 81 and 82 to record the data and their second mating led to pregnancy. To check the fertility of the male whose first pairing did not result in mating, male no. 28 from the 300-mg/kg bw group was mated a second time with one female from a reserve group and mating led to pregnancy.

Reproduction data:
No treatment-related differences from the control group were recorded in the mean of implantation sites per litter or corpora lutea.
No treatment-related differences from the control group were recorded in the percentage of pre- or postimplantation losses. Female no. 45 from control group showed 100% implantation site loss. No differences in sex ratio were recorded.

Breeding data:
The length of pregnancy was similar in all groups with a mean of 21-22 days.
Female no. 78 at 1000 mg/kg bw had not enough milk and did not nurse correctly its pups; consequently, the pups died or were devoured between days 2 and 3 of lactation. Despite the fact that the pups had milk in the stomach when alive, they did not have any at necropsy and showed hypothermia and pallor. Female no. 78 showed slightly enlarged mammary glands. Even the percentage of pups showing milk in the stomach was similar in all groups. One pup in the litter of females no. 43 from control group, no. 63 at 300 mg/kg bw, no. 78 and two pups from no. 73 at 1000 mg/kg bw showed no milk in stomach on day 1 postpartum and died.

ORGAN WEIGHTS (PARENTAL ANIMALS)
At 1000 mg/kg bw, kidney weights tended to be slightly higher in males. These differences were not statistically significant. Moreover, lower pituitary and adrenal weights were recorded; the differences were statistically significant for pituitary in males. A trend to higher liver weight was recorded in females at 1000 mg/kg bw, but the differences were not statistically significant.
Concerning reproductive organs, lower left ovary weights were recorded at 1000 mg/kg bw. This difference was statistically significant in relation to body weight ratio.

GROSS PATHOLOGY (PARENTAL ANIMALS)
At 1000 mg/kg bw, large right kidney was observed in one male and reddish thymus in another one. A reddish area on cecum wall was observed in one male and reddish foci in lungs in another one. Likewise, thickened gastric mucosa was observed in one female and small spleen in another one.
At 300 mg/kg bw, reddish foci were observed in the thymus of one male. No findings were observed in females.
At 100 mg/kg bw, pale kidneys were observed in one male and reddish foci in lungs in another one. Likewise, left seminal vesicle reduced in size was observed in one male and reddish thymus in three males. Thickened gastric mucosa was observed in one female.
In the control group, one male had left seminal vesicle reduced in size. One male and one female had pale kidneys. In addition, one male had pancreas reduced in size and thymus with reddish foci.
Yellowish-whitish gastric mucosa was recorded in few females from all groups (including control group).

HISTOPATHOLOGY (PARENTAL ANIMALS)
All microscopic findings recorded were considered to be within the range of normal background lesions that may be seen in rats of this strain and age and under the experimental conditions used in this study.
Key result
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed in the study
Key result
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed in the study
Key result
Critical effects observed:
no
Clinical signs:
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
No treatment-related differences from the control group were recorded in the mean of dead pups at the first litter check and living pups. During the first 4 days postpartum, the percentage and mean of postnatal losses was slightly higher at 1000 mg/kg bw; however the number of litters affected was similar to that of the control group (2 vs 1 litters) due to the fact that female no. 78 (1000 mg/kg bw) lost all its litter. The resulting viability index was lower than in the control group (84.7% versus 99.1%, respectively).
No differences were recorded at 100 and 300 mg/kg bw.


BODY WEIGHT (OFFSPRING)
No differences were recorded between groups and sexes. At 1000 mg/kg bw, one runt pup *no.7 from litter no.54) was observed.

GROSS PATHOLOGY (OFFSPRING)
No noteworthy findings were recorded in the control, 100, 300 or 1000 mg/kg bw pups.

MORPHOLOGICAL EXAMINATION
No treatment-related alterations were recorded in the morphological examination of the pups.
One pup from control group had a hematoma on toes and one pup from 300 and 1000 mg/kg bw group had a wound on scapula or limbs.

OTHER FINDINGS (OFFSPRING)
Motor development: Regarding postural reflexes in the pups, a significantly lower percentage of fetuses with positive response in the surface-righting reflex (righting reflex) at 1000 mg/kg bw was recorded compared to the control group. There were no treatment-related differences compared to the control animals at 100 or 300 mg/kg bw.
Key result
Dose descriptor:
NOAEL
Remarks:
developmental
Generation:
F1
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed in the study
Key result
Critical effects observed:
no
Key result
Reproductive effects observed:
no
Conclusions:
Docosyl docosanoate (CAS No 17671-27-1), a structural analogue, was tested for its potential to induce developmental and/or reproductive toxic changes in rats in an OECD 422 study. Based on the the lack of adverse effects the NOAEL (systemic, reproduction, developement) was derived at > 1000 mg/kg bw/d.
Executive summary:

Docosyl docosanoate (CAS No 17671-27-1), a structural analogue, was tested for its potential to induce developmental and/or reproductive toxic changes in rats in an OECD 422 study. Groups of 10 male and 10 female rats were dosed with 0, 100, 300 and 1000 mg/kg bw/d.

Observations in parental animals:

No mortality and no relevant clinical signs were observed. There were no test-item-related differences from the control group were recorded for body weight. All sperm parameters recorded were within the range of normal background lesions which may be recorded in animals of this strain and age. No chances were observed regarding the reproductive performance.

Additinally, no relevant changes in organ weights and gross pathology/histopathology were reported.

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:
Reliable without restriction
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available

Effects on developmental toxicity

Description of key information

not madatory

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

Toxicity to reproduction: other studies

Description of key information

no further relevant data available

Mode of Action Analysis / Human Relevance Framework

In the absence of any evidence for species specific effects or modes of action the effects observed in animals are regarded as relevant for humans.

Justification for classification or non-classification

Due to the lack of reproductive and develpomental toxic effects in OECD 422 repeated dose toxicity studies in rats the substance does not have to be classified for developmental and reproductive toxicity according to the criteria laid down in the Regulation (EC) No 1272/2008.

Additional information