Registration Dossier

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because a pre-natal developmental toxicity study is available
other:
Cross-reference
Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
July - October 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
The deviations were considered not to affect the outcome or integrity of the study (see "Any other information on materials and methods incl. tables")
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, Italy
- Age at study initiation: 9 weeks old
- Weight at study initiation: 219 - 296 g
- Fasting period before study: no
- Housing: singly housed in appropriately sized, suspended cages with stainless steel grid tops and solid bottoms, containing an integral food hopper, bedding material: sterilised white wood shavings
- Diet: SDS Rat and Mouse (modified) No. 3 Diet SQC Expanded ad libitum
- Water: water from the public supply ad libitum (analysed at regular intervals for dissolved material, heavy metals, pesticide residues, pH, nitrates and nitrites, microbiological screening was also conducted)
- Acclimation period: 3-5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 40-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2013-07-26 To: 2013-08-14
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The required amount of test item was weighed and accurately transferred to a pre-labelled container. The appropriate amount of the vehicle was added to the container and magnetically stirred until a homogenous formulation was obtained.
The dosing formulation were pepared weekly, stored at 2-8°C in the dark for up to 8 days, and dispensed daily. The dosing formulations were stirred for at least 30 minutes before and continuously during dosing.

VEHICLE
- Justification for use and choice of vehicle (if other than water): no justification given
- Concentration in vehicle: 0 / 40 / 200 / 400 mg/mL
- Amount of vehicle (if gavage): 2.5 mL/kg
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analyses were performed by Gas Chromatography with Flame Ionisation Detection using a validated analytical procedure. Samples were analysed from formulations prepared for use in Week 1, 2 and 3. Samples for analysis were taken in duplicate from the top, middle and bottom of the dosing solutions, backup samples were taken in triplicate (top, middle and bottom). Analysis were done for concentration and homogeneity. The homogeneity results obtained from the top, middle and bottom for the test group preparations were averaged and utilised as the concentrations results.
Acceptance criteria for concentration: mean sample concentration results within or equal to ± 10% of theoretical concentration, each individual sample concentration result within of equal to ± 15%. Acceptance criteria for homogeneity: relative standard deviation of concentrations of ≤ 10% for each group.
All formulations from weeks 2 and 3 and group 1-3 formulations (control, low and middle doses) from week 1 were found to be within the acceptance criteria of the analytical method. Group 4 (high dose) on week 1 were found to be -1.8% below the acceptance criteria, however, due to the small degree of the discrepancy, this was not considered to have affected the outcome of the study. There was a low coefficient of variation (≤ 6.6%) in the analyses indicating that the formulations were homogenous.
Details on mating procedure:
- Impregnation procedure: purchased timed pregnant
Duration of treatment / exposure:
from days 6-19 of gestation
Frequency of treatment:
once daily
Duration of test:
20 days
Remarks:
Doses / Concentrations:
100 mg/kg/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
500 mg/kg/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg/day
Basis:
actual ingested
No. of animals per sex per dose:
24
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: the dose levels were chosen based on previous toxicity studies including a 14 day dose range finding study (Charles River Laboratories Study No. 523891) and a 90 day repeat dose study (Charles River Laboratories Study No. 523907).
Results of 14 day dose range finding study:
The objective of this study was to determine the potential toxicity of Coconut Oil, Reaction Products with Polyethylene Glycol and Trimethylolpropane when given by oral gavage for 14 days to Han Wistar rats. These data provided information to allow the selection of suitable dosages for further repeat dose studies.
The study design was as follows: 5 animals per Sex and group, dose levels: 0, 100, 500 and 1000 mg/kg bw/day. Coconut Oil, Reaction Products with Polyethylene Glycol and Trimethylolpropane was administered using corn oil as the vehicle and a constant dose volume of 2.5 mL/kg. The following parameters and end points were evaluated in this study: clinical signs, body weights, body weight changes, food consumption, and on Day 15, gross necropsy findings and organ weights. There were no unscheduled deaths or adverse clinical signs that were considered to be related to treatment. Body weights, food consumption and organ weights were considered to be unaffected by treatment. In conclusion, there was no evidence of toxicity after 14 days of oral gavage administration of 1000 mg/kg/day of Coconut Oil, Reaction Products with Polyethylene Glycol and Trimethylolpropane to Han Wistar rats. Based on these results, 1000 mg/kg/day was considered to be a suitable high dose level for further repeat dose studies in this species using this test item.
For details of the 90 day repeat dose study see cross-reference.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes, animals checked for viability
- Time schedule: early morning and as late as possible each day

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: once at day 3 of gestation (pretrial), daily at days 6 - 20 of gestation (dosing period)

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

WATER CONSUMPTION AND COMPOUND INTAKE: Yes, on a regular basis throughout the study, monitored by visual inspection of the water bottles. Water consumption was not assessed by weight, as there were no inter group differences.


POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: carcass and muscoloskeletal system, external surfaces and orfices, cranial cavity and external surfaces of the brain, thoracic, abdominal, and pelic cavities with their associated organs and tissues

OTHER:
Pre/postdose observations: prior to dosing and regularly throughout the study, including examination for reaction to treatment, particular attention was paid to the animals during and for the first hour after dosing
Ovaries and uterine content:
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, each implant was classified as being live, or a dead fetus (dead full term fetus that shows no sign of maceration)
- Number of early resorptions: Yes (discrete, formless, discoloured tissue mass attached to the internal uterine wall; may be varying in size)
- Number of late resorptions: Yes (macerated tissue identifiable as an embryo fetus, with recognizable external features such as tail, limbs, mouth and nares present; attached to distinct identifiable placenta)
- Other:
Placentae (size, color, shape), any abnormalities were recorded
Number and distribution of live and dead fetuses
Fetal examinations:
- External examinations: Yes, all per litter (late death and dead fetuses were examined to the extent possible)
- Soft tissue examinations: Yes, half per litter fixed in Bouin's fluid, examined for soft tissue abnormalities and for sex (Wilson's freehand sectioning technique), the other half of the litter fixed in methylated ethyl alcohol and examined by open dissection for abnormalities of the thoracic and abdominal viscera, and for sex.
- Skeletal examinations: Yes, half per litter (the eviscerated carcasses, fixed in methylated ethyl alcohol, were macerated in potassium hydroxide, the skeletons stained with Alizarin Red S, and then cleared with aquaeous glycerol solutions, the preparations were examined for the presence of skeletal abnormalities and the extent of ossification)
- Head examinations: No data
Statistics:
Means and standard deviations were calculated for body weight, food consumption and pregnancy data.
Body weight and food consumption, data were analysed for homogeneity of variance using the F-Max test. If the group variances appear homogeneous, a parametric ANOVA was used and pairwise comparison were made using Fisher's F protected LSD method via Student's test ie pairwise comparisons were made only if the overall F-test was significant. If the variances were heterogenous, log or square root transformations were used in an attempt to stabilise the variances. If the variances remain heterogeneous, then a Kruskal-Wallis non-parametric ANOVA was used and pairwise comparisons were made using chi squared protection (via z tests, the non-parametric equivalent of Student's test).
Historical control data:
Historical control data on pre-implantation loss were included in the report. The pre-implantation loss in 12 studies performed in the testing laboratory ranged from 2 - 24 % (see table "Historical Control Data: Pre-implantation loss" in Overall remarks, attachments
Clinical signs:
no effects observed
Description (incidence and severity):
No treatment related effects observed.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
No premature deaths on this study.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
At 500 mg/kg/dday and above there was an apparent slight decrease in group mean body weight gain, throughout the dosing perio. However, it is noted on Day 1 of dosing that the group mean body of animals in groups 1 and 2 were slghtly higher than the group mean body weight of animals in groups 3 and 4. This slight difference in body weight did not achieve statistical significance and was not positively attributed to treatment in the absence of any other findings.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Any achieved statistical significance was sporadic and no dose related pattern of effect could be established and so these differences were considered not to be associated with treatment.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
The type and distribution of gross pathology findings did not suggest any adverse effects of treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
no effects observed
Number of abortions:
no effects observed
Pre- and post-implantation loss:
effects observed, non-treatment-related
Description (incidence and severity):
At 1000 mg/kg bw/d there was an apparent increase in the group mean pre-implemenation loss, when compared to Controls. The group mean implantation loss is 10 % at 1000 mg/kg bw/d compared to 3 % in controls. At 100 and 500 mg/kg bw/d there was a very marginal increase in totol dead implants when compared to control. The total dead implants increased from 6 % in Controls to 7 % and 8 %, respectively, at 100 and 500 mg/kg bw/d. Although this appears to increase in a dose dependent manner, it is noted that the difference in total dead implants are too small to be positively attributed to treatment.
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
effects observed, non-treatment-related
Description (incidence and severity):
There was a higher than expected number of non-pregnant animals in all groups, with the highest number in Group 2 (5/24 animals not pregnant). As these non-pregnant animals were distributed evenly throughout the control and dose groups, this was considered not to be related to treatment.
Other effects:
no effects observed
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
- Mortality: No premature deaths occurred
- Clinical observations: At the high dose group, one animal was found to have ploughing behaviour and salivation on Day 16 of gestation only (immediately post dose). As this was an isolated incident in one animal only this was not considered to be adverse. For details see table 1 below.
- Body weights: At 500 mg/kg/day and above was an apparent slight decrease in group mean body weight gain, throughout the dosing period. However, it was noted on Day 1 of dosing that the group mean body weight of animals in the control and low dose group were slightly higher than the group mean body weight of animals in the mid and high dose groups. This slight difference in body weight did not achieve statistical significance and was not positively attributed to treatment in the absence of any other findings. For details see table 2 below.
- Food consumption: The group mean food consumption performance in all dose groups was similar to controls. For details see table 3 below.
- Gross pathology: The type and distribution of gross pathology findings did not suggest any adverse effects of treatment. For details see table 1 below.
- Pregnancy performance: There was a higher than expected number of non-pregnant animals in all groups, with the highest number in the low dose group. (5/24 animals not pregnant). As these non-pregnant animals were distributed evenly throughout the control and dose groups, this was considered not to be related to treatment.
At 1000 mg/kg/day there was an apparent increase in the group mean pre-implantation loss, when compared to controls (high dose group: 10%, controls 3%). At the test facility pre-implantation losses of up to 24% have been observed in this species and strain (historical control data, see table below). Also due to a lack of any other treatment related effects in the dam of the developing fetus, the weight of evidence suggests that this difference in pre-impantation loss is not related to treatment and is due to natural variation within this species and strain.
At 100 and 500 mg/kg/day there was a very marginal íncrease in total dead implants, when compared to control. The total dead implants increased from 6% in control to 7% and 8%, respectively, at 100 and 500 mg/kg/day. Although this appears to increase in a dose dependent manner, it is noted that the difference in total dead implants are too small to be positively attributed to treatment.
For details see table 4 below.
Key result
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Key result
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Description (incidence and severity):
It is noted that Animals 22 (Group 1) and Animals 68 and 71 (Group 3) had fetuses much larger than expected for Day 20 of gestation. It is assumed that there was an error during the mating of these animals at the supplier and they were actually on a later day of gestation. Therefore the fetal weights and gravid uterus weights of these animals have been excluded from group calculations.
The group mean fetal weights in all dose groups are similar to controls.
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Description (incidence and severity):
The type and distribution of fetal abnormalities, variants and skeletal ossification parameters did not indicate any relationship to treatment with Coconut oil, reaction products with polyethylene glycol and trimethylolpropane. Any slight intergroup differences were considered to be due to intergroup variation and too small to be attributed to treatment.
At 100 mg/kg/day and above several fetuses were found to have eye(s) oval in shape. This abnormality was found in 2, 3 and 1 fetus in Groups 2, 3 and 4, respectively. Due to the low overall incidence of this abnormality and due to a lack of treatment related response throughout the dose groups, this could not be positively related to treatment.
Visceral malformations:
no effects observed
Other effects:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
- Fetal weights: The group mean fetal weight in all dose groups were similar to controls. For details see table 4 below. (1 animals of the control group and 2 animals of the mid dose group had fetuses much larger than expected for Day 20 of gestation. It is assumed that there was an error during the mating of these animals at the supplier and they were actually on a later day of gestation. The fetal weight of these animals were excluded from group calculations.)
- Fetal abnormalities and variants: The type and distribution of fetal abnormalities, variants and skeletal ossification parameters did not indicate any relationship to treatment with the test substance. At the low dose group and above several fetuses were found to have eye(s) oval in shape (2, 3 and 1 fetus in the low, mid and high dose group, respectively). Due to the low overall incidence of this abnormality and due to a lack of treatment related response throughout the dose groups, this could not be positively related to treatment. For details see tables 5, 6 and 7 below.
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no
Table 1: Group Incidence of Clinical Observations and Necropsy Findings
Dose levels [mg/kg/day]
0 100 500 1000
Clinical Observations:
Sparse hair 4 3 3 4
Staining on fur: head / dorsal neck / ears / around vagina 7 4 2 1
Damaged toe 0 1 0 0
scapb(s): foot/ feet / ventral abdomen / dorsal neck 0 3 1 2
Teeth damaged / overgrown 0 1 0 0
Foot enlarged 0 0 1 0
Ploughing 0 0 0 1
Excess salivation 0 0 0 1
Abnormal colour discharge, vagina 0 0 0 1
Necropsy findings:
Staining on fur: head / dorsal neck / ears / around vagina 4 3 2 1
Thin hair coat 0 2 1 2
Scab(s): dorsal neck 0 1 1 2
Liver enlarged 0 0 1 0
Liver, area pale 0 0 1 0
Spleen enlarged 0 0 1 0
Number of females 24 24 24 24
Table 2: Body Weights, Pregnant Animals only
Day of gestation Dose levels [mg/kg/day]
0 100 500 1000
Group mean values± SD
[g]
Group mean values± SD
[g]
Group mean values± SD
[g]
Group mean values± SD
[g]
3 238 ± 17 240 ± 16 234 ± 13 231 ± 19
6 261 ± 15 261 ± 17 255 ± 12 253 ± 18
7 264 ± 15 266 ± 18 260 ± 12 257 ± 18
8 270 ± 14 271 ± 18 263 ± 13 262 ± 19
9 276 ± 14 277 ± 18 270 ± 14 267 ± 19
10 281 ± 15 284 ± 17 275 ± 15 273 ± 20
11 291 ± 16 292 ± 18 285 ± 15 281 ± 22
12 298 ± 16 300 ± 18 292 ± 13 289 ± 23
13 307 ± 15 208 ± 19 299 ± 16 295 ± 23
14 315 ± 17 314 ± 21 308 ± 16 301 ± 24
15 324 ± 16 322 ± 23 317 ± 16 311 ± 25
16 334 ± 17 332 ± 24 326 ± 18 321 ± 26
17 347 ± 18 344 ± 26 339 ± 19 331 ± 27
18 361 ± 21 359 ± 28 256 ± 23 346 ± 30
19 375 ± 21 371 ± 30 364 ± 24 357 ± 29
20 378 ± 24 375 ± 30 365 ± 26 361 ± 29
Change 6-20 117± 17 115 ± 23 110 ± 24 108 ± 18
significantly different from control group: * = p<00.05, ** = p<0.01, *** = p<0.001
Table 3: Food Consumption (g/animal/day), Pregnant Animals only
Day of gestation Dose levels [mg/kg/day]
0 100 500 1000
Group mean values± SD
[g]
Group mean values± SD
[g]
Group mean values± SD
[g]
Group mean values± SD
[g]
4 22.9 ± 2.4 23.8 ± 3.1 22.3 ± 3.2 22.5 ± 3.2 #
5 25.4 ± 3.7 24.4 ± 4.0 24.1 ± 3.0 24.4 ± 3.1
6 26.4 ± 3.1 26.1 ± 2.9 24.7 ± 2.3 25.1 ± 3.2
7 22.3 ± 4.0 23.0 ± 2.6 22.0 ± 4.2 23.0 ± 3.4
8 22.9 ± 2.4 23.0 ± 2.6 21.5 ± 2.4 22.7 ± 2.8
9 23.9 ± 2.9 24.1 ± 4.1 21.9 ± 3.1 21.6* ± 3.6
10 26.2 ± 5.6 25.9 ± 4.2 24.0 ± 5.2 24.2 ± 4.6
11 27.1 ± 3.4 27.2 ± 3.5 26.1 ± 3.5 25.2 ± 4.3
12 27.1 ± 2.6 26.8 ± 3.0 25.6 ± 4.0 26.5 ± 3.7
13 28.1 ± 3.0 28.1 ± 3.9 25.9 ± 4.0 25.9 ± 4.5
14 27.9 ± 2.6 25.8 ± 3.8 26.4 ± 2.7 25.6 ± 3.8
15 27.8 ± 2.6 27.1 ± 3.5 25.2 ± 3.4 26.9 ± 3.5
16 27.3 ± 3.4 26.7 ± 4.8 26.6 ± 3.5 26.0 ± 3.9
17 27.3 ± 3.1 26.5 ± 3.2 26.4 ± 5.6 26.4 ± 3.1
18 28.8 ± 5.8 30.6 ± 5.7 27.5 ± 6.2 27.2 ± 4.9
19 27.2 ± 5.8 26.9 ± 6.2 22.0** ± 6.4 25.8 ± 5.6
20 17.2 ± 6.1 17.6 ± 5.8 13.5 ± 6.7 17.2 ± 5.2
significantly different from control group: * = p<00.05, ** = p<0.01, *** = p<0.001
# Group mean value from 20 pregnant animals
Table 4: Pregnancy Performance and Fetal Weights
Dose levels [mg/kg/day]
0 100 500 1000
Number of animals mated 24 24 24 24
Number of non-pregnant animals* 4 5 2 3
Number pregnant at Day 20 necropsy 20** 19 22 21
Pregnancy frequency 79% 79% 92% 88%
Total corpora lutea graviditatis 281 262 308 287
Total number of implants 272 248 294 257
Pre-implantation loss 3% 5% 5% 10%
Total live implants 255 (94%) 230 (93%) 271 (92%) 247 (96%)
Total dead implants 17 (6%) 18 (7%) 23 (8%) 10 (4%)
Total early embryonic deaths 15 (6%) 18 (7%) 22 (7%) 10 (4%)
Total late embryonic deaths 2 (1%) 0 1 (0.3%) 0
Total fetal deaths 0 0 0 0
Mean corpora lutea graviditatis 14.8 ± 2.3 13.8 ± 4.3 14.0 ± 2.2 13.7 ± 2.9
Mean implants 14.3 ± 2.6 13.1 ± 4.4 13.4 ± 3.0 12.2 ± 3.8
Mean live implants 13.4 ± 2.8 12.1 ± 4.1 12.3 ± 3.4 11.8 ± 3.6
Mean dead implants 0.9 ± 0.9 0.9 ± 1.2 1.0 ± 1.4 0.5 ± 0.8
Mean early embryonic deaths 0.8 ± 0.8 0.9 ± 1.2 1.0 ± 1.3 0.5 ± 0.8
Mean late embryonic deaths 0.1 ± 0.3 0 0.05 ± 0.2 0
Mean fetal deaths 0 0 0 0
Total live male fetuses 134 (53%) 114 (50%) 149 (55%) 125 (51%)
Total live female fetuses 121 (47%) 116 (50%) 122 (45%) 122 (49%)
Live fetal sex ratio (male : female) 1 : 0.90 1 : 1.02 1 : 0.82 1 : 0.98
Mean total uterus weight [g] 83± 15 75 ± 23 78 ± 18 72 ± 20
Mean litter mean fetal weight [g] 3.96 ± 0.26 3.96 ± 0.31 3.76 ± 0.36 3.95 ± 0.41
* Non-pregnant animals excluded from further assessment
** includes one animal which had only one late death implant at necropsy, animal was excluded from further assessment
Table 5: Group Incidence of Major Fetal Abnormalities   
Abnormality Dose levels [mg/kg/day]
0 100 500 1000
Incidence of Fetuses (Litters)
Subcutaneous mass fore/midbrain region of head with marked subdural haemorrhage, with/without haemorraghe within lateral brain ventricles. 0 0 2(1) 0
Tympanic annuli incompletely ossified. Scapulae bent. Rib(s) kinked. Radii bent. Femur shortened and bent. 1(1) 0 0 0
Cervical vertebral arches connected. Thoracic vertebra hemicentric. Split sternum. 1(1) 0 0 0
Number with major abnormality 2(2) 0 2(1) 0
Total number examined 255(19) 230(19) 271(22) 247(21)
Table 6: Group Incidence of Minor Fetal Abnormalities and Variants
Abnormality/Variant Dose levels [mg/kg/day]
0 100 500 1000
Incidence of Fetuses (Litters)
Visceral:
Subcutaneous haemorraghe:
Head 0 0 0 2(2)
Trunk 0 1(1) 0 1(1)
Increased subdural space, brain ventricel dilated 0 0 1(1) 0
Eye(s) oval (in shape) 0 2(2) 3(1) 1(1)
Eye lens oval (in shape) 0 0 1(1) 0
Thyroid markedly reduced (in size) 2(1) 0 0 0
Cervical remnant of thymus 10(6) 10(7) 4(3) 4(3)
Innominate artery absent 1(1) 0 1(1) 0
Tendinous region of diaphragm locally thinned with minimal protrusion of median liver lobe 4(4) 3(3) 2(2) 3(2)
Tendinous region of diaphragm locally thinned with protusion of median liver lobe 0 0 1(1) 2(2)
Median liver lobe bifurcated 0 0 1(1) 0
Additional liver lobe within median cleft 14(10) 15(10) 7(7) 16(10)
Hepatic haemorraghe 2(2) 2(1) 0 0
Minimal intra-abdominal/intra-abdominal haemorraghe 2(2) 0 1(1) 0
Renal pelvis dilated 1(1) 0 1(1) 2(2)
Testis(es) not fully descended to pelvic position 1(1) 0 0 1(1)
Testis(es) medially displaced 1(1) 0 1(1) 0
Urinary bladder enlarged 0 1(1) 1(1) 0
Umbilical artery left-sided 1(1) 2(2) 0 2(2)
Genital papilla elongated 0 1(1) 0 0
Fetus(ses) larger than normal for day 20 of gestation 3(1) 0 10(2) 0
Small fetus 1(1) 1(1) 0 0
Number with minor abnormality/variant 41(17) 34(15) 33(16) 32(17)
Number examined by Wilson section 126(19) 115(18)# 135(22) 124(21)
Skeletal:
Cranial bone(s) small linear ossification irregularity/ies 1(1) 2(2) 0 1(1)
Cranial bone(s) small discrete unossified/incompletely ossified area(s) 1(1) 1(1) 0 0
Additional ossified arear(s) between cranial bone(s) 0 0 0 1(1)
Jugal(s) connected/fused to zygomatic process of maxilla 4(1) 0 0 0
Presphenoid incompletely ossified 0 0 4(2) 1(1)
Cervical vertebral arch(es) increased ossification 7(6) 10(9) 9(7) 7(4)
Cervical rib(s) 2(2) 1(1) 0 1(1)
Additional ossified area arising from sternebra 1(1) 2(2) 2(2) 2(2)
Rib(s) minimally kinked 2(2) 1(1) 3(3) 1(1)
Rib(s) incompletely ossified 1(1) 1(1) 1(1) 0
Rib(s) costal cartilages asymmetrically aligned 3(3) 2(2) 6(5) 2(2)
Rib(s) costal artilage(s) not attached to sternum 7(4) 1(1) 2(2) 3(3)
Pelvic girdle bilateral cranial displacement 0 1(1) 0 0
Number with minor abnormality/variant 25(12) 19(12) 26(16) 18(11)
Total number examind skeletally 129(19) 115(19) 136(22) 123(21)
Number of ribs:
13th vestigial rib(s) 1(1) 1(1) 1(1) 2(2)
13th reduced rib(s) 11(5) 4(4) 9(8) 15(8)
13 complete rib(s) 107(19) 102(19) 116(22) 201(21)
Vestigial supernumerary rib(s) on 1st lumbar vertebra 10(6) 5(2) 10(5) 4(3)
# One litter with only one fetus examined skeletally, no fetus for Wilson sectioning
Table 7: Group Incidence of Skeletal Ossification Parameters
Parameter Dose levels [mg/kg/day]
0 100 500 1000
Incidence of Fetuses (Litters)
Incomplete ossification affecting:
≥4 skull bones 4(2) 4(3) 7(5) 4(3)
≤3 skull bones 30(14) 24(12) 32(18) 33(14)
Cervical vertebral arch(es) 2(2) 1(1) 3(2) 1(1)
Thoracic vertebral centrum(s) 6(6) 6(6) 2(2) 6(4)
Lumbar vertebral arch(es) 1(1) 0 0 0
Pubis(es) 3(3) 6(5) 6(4) 5(4)
Ischium(s) 0 0 2(2) 0
Sacral vertebral arch(es) 6(5) 4(3) 10(6) 10(5)
2nd metacarpal(s) 0 1(1) 1(1) 2(2)
Unossified:
Hyoid 2(2) 0 0 0
5th metacarpal(s) 22(9) 39(11) 30(13) 36(14)
5th metatarsal(s) 1(1) 0 1(1) 1(1)
Ossified:
Anterior arch of atlas 54(17) 29(11) 44(17) 46(17)
>2 cervical vertebral centra 16(9) 12(7) 14(6) 13(8)
One of more sacrocaudal vertebrae with connection between centrum and arch(es) 53(16) 35(14) 38(16) 55(16)
Phalangeal elements 17(5) 15(7) 28(8) 25(10)
1st metatarsal(s) 2(1) 0 5(2) 3(1)
Calcaneum 0 0 4(1) 0
Mean number of caudal vertebral centra 4.2 4.0 4. 4.2
Number of sternebae retarded:
0 68(19) 44(15) 54(17) 48(18)
1 48(15) 47(16) 49(19) 45(18)
2 10(6) 21(11) 21(11) 25(10)
>2 3(3) 3(3) 12(8) 5(3)
Total number examined skeletally 129(19) 115(19) 136(22) 123(21)
Conclusions:
Under the conditions of this study, the maternal and fetal no observed effect level (NOEL) was considered to be 1000 mg/kg bw/day.
Executive summary:

The effects of Coconut oil, reaction products with polyethylene glycol and trimethylolpropane, administered orally during the period of organogenesis and fetal development in pregnant rats was examined according to OECD guideline 414 and under the regulations of GLP.

Time-mated female Sprague-Dawley rats were randomised into 3 test groups and 1 control group (24 animals per group), receiving 100, 500 and 1000 mg/kg bw test substance and vehicle (corn oil), respectively, once daily by oral gavage. All animals were dosed over Days 6 -19, inclusive of gestation, where the day of detection of mating was designated Day 0. Animals were regularly monitored for clinical signs of toxicity, body weight and food consumption performance and were killed on Day 20 of gestation for examination of pregnancies and embryo-fetal development.

At levels of up to 1000 mg/kg bw/day there were no clinical observations that could be associated with treatment and there was no effect of treatment observed on body weight gain, or food consumption performance.

At 100 and 500 mg/kg bw/day there was a very marginal increase in total dead implants, when compared to control (6% in controls, 7% and 8%, respectively, at 100 and 500 mg/kg bw/day). Although this appears to increase in a dose dependent manner, it was noted that the difference in total dead implants were too small to be positively attributed to treatment.

At 1000 mg/kg bw/day there was an apparent increase in pre-implantation loss (10% compared to 3% in controls). At the test facility pre-implantation losses of up to 24% have been observed in this species and strain. Also due to lack of any other treatment related effects in the dam or the developing fetus, the weight of evidence suggests that this difference in pre-implantation loss is not related to treatment and is due to natural variation within this species and strain.

At levels up to 1000 mg/kg bw/day there were no differences in pregnancy performance that could be positively attributed to treatment with the test substance.

Throughout the dose groups there was a higher than expected number of non-pregnant animals along with several animals which were suspected as being on the incorrect day of gestation. These animals have been excluded from the majority of the results; nonetheless, we have the following number of evaluable dams/litters per group: Group 1- 19 dams/litters, Group 2- 19 dams/litters, Group 3- 20 dams/litters and Group 4- 21 dams/litters. The OECD 414 guideline recommends approximately 20 females per group with implantation sites at necropsy and that groups with fewer than 16 animals with implantation sites may be inappropriate. As all groups in this study have more than 16 evaluable pregnant females, it is considered that this is enough animals to provide sufficient data for the study and to satisfy the recommendations of the OECD 414 guideline.

The type, incidence and distribution of fetal abnormalities and variations and skeletal ossification parameters did not indicate any association with treatment.

Under the conditions of this study, the maternal and fetal no observed effect level (NOEL) was considered to be 1000 mg/kg bw/day.

Data source

Materials and methods

Results and discussion

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion