Registration Dossier

Diss Factsheets

Toxicological information

Developmental toxicity / teratogenicity

Currently viewing:

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
publication
Title:
Developmental toxicity evaluation of trimethylolpropane caprylate caprate in Sprague-Dawley rats.
Author:
Azuka, C. and Daston, P.
Year:
2004
Bibliographic source:
Birth Defects Res B Dev Reprod Toxicol. 2004 Dec;71(6):374-9.

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
few details on test substance given, no analysis of the test compound
GLP compliance:
not specified
Limit test:
no

Test material

Constituent 1
Reference substance name:
Decanoic acid, ester with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol octanoate
EC Number:
234-392-1
EC Name:
Decanoic acid, ester with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol octanoate
Cas Number:
11138-60-6
IUPAC Name:
2,2-bis[(octanoyloxy)methyl]butyl decanoate

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Kingston, NY
- Age at study initiation: young adult
- Weight at study initiation: Mean of the maternal body weight: 226 g (Vehicle), 225 g (200 mg/kg bw/day), 227 g (600 mg/kg bw/day), 226 g (2000 mg/kg bw/day)
- Fasting period before study: No
- Housing: Virgin females were cohabitated with singly-housed male rats, one male per female rat for a maximum of 5 days and returned to individual housing in stainless steel wire-bottomed cages after mating.
- Diet: Certified Rodent Diet No. 5002 (PMI Feeds Inc. St.Louis, MO), ad libitum
- Water: water passaged through a reverse osmosis membrane with chlorine added as a bacteriostat, ad libitum
- Acclimation period: yes, period not mentioned

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25
- Humidity (%): 30 - 70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:
dermal
Vehicle:
corn oil
Details on exposure:
TEST SITE
- Area of exposure: The back of the animals from the shoulders to the hip joints and extended ventrolaterally from the dorsal midline on each side (5x7 cm)
- % coverage: approx. 10% of the body surface
- Type of wrap: occlusive, gauze pad secured with Vetrap or Micropore tape
- Time intervals for shavings or clippings: during acclimatization period

REMOVAL OF TEST SUBSTANCE
- Washing: exposed area was wiped with a dermal wipe pad dampened with aqueous 1% solution of soap and then patted dry with a second clean pad
- Time after start of exposure: 6 h

TEST MATERIAL
- Amount(s) applied: 2 mL/kg
- Concentration: 200, 600, and 2000 mg/kg/day
- Constant volume or concentration used: yes

VEHICLE
- Amount(s) applied: 2 mL/kg
- Concentration: up to 100% (vehicle control)

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes, Elizabethan collar
Analytical verification of doses or concentrations:
no
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: maximum 5 days
- Further matings after two unsuccessful attempts: Not applicable
- Verification of same strain and source of both sexes: No Data
- Proof of pregnancy: Both, vaginal plug and/or sperm in vaginal smear were referred to as Day 0 of pregnancy
Duration of treatment / exposure:
Treatment on Gestation Days (GD) 6 - 15
Frequency of treatment:
Daily
Duration of test:
Termination of the study by CO2 inhalation on GD 20.
No. of animals per sex per dose:
25
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose dependent occurrence of skin irritation. Higher levels than 2000 mg/kg bw/day might be expected to produce marked irritation thereby compromising the interpretaion of developmental results.
- Rationale for animal assignment (if not random): Computer-generated randomization by weight (Barlett´s test for homogeneity) such that the groups were not statistically different (5% significance level) from each other.

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Animals were checked for mortality twice daily during the treatment period and daily thereafter.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Animals were checked for signs of reaction to treatment and/or symptoms of illness once daily before treatment, approx. 60 min after treatment during the dosing period. The dosing site was examined daily prior to substance application for signs of skin irritation according to Draize.

BODY WEIGHT: Yes
- Time schedule for examinations: Recorded on GD 0 and daily during the treatment period.

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

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day 20
- Organs examined: The uterus, uterine contents, position of the fetuses in the uterus and number of corpora lutea. Number and distribution of intrauterine implantations were classified as live or death fetuses, late intrauterine deaths (resorptions) and early intrauterine resorption sites. Live fetuses were sexed and further examined (see below).
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
- Number of early resorptions: Yes
- Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter (the heads of the animals used for soft tissue examinations)
Statistics:
Clinical observations and other proportion data were analyzed using the Variance Test for Homogeneity of the Binominal Distribution. Quantitative continuous data were analyzed using Barlett´s Test for Homogeneity of Variance and the Analysis of Variance when Barlett´s Test was not significant (p >0.05). If the Analysis of Variance was significant (p>0.05), Dunnett´s Test was used to identify the statistical significance of the individual groups. If the Analysis of Variance was not appropriate, the Kruskal-Wallis Test was used when >75% ties were present. In case of significance (p >0.05), Dunn´s Method of Multiple Comparisons was used for identification of statistical significance of the individual groups.
Historical control data:
No details. One dam having a litter consisting of seven early resorptions was pointed out as single non-dosage dependent event and to be within the ranges observed historically at the test facility.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
no effects observed
Dermal irritation (if dermal study):
effects observed, treatment-related
Description (incidence and severity):
The two highest dose levels caused some local irritation at the site of application, but no decreases in maternal weight gain or feed consumption (non-adverse)
Mortality:
mortality observed, non-treatment-related
Description (incidence):
Two animals in the control group and one animal in the high-dose group died within 6 h after first application; these were not considered to be treatment related and the animals were replaced (non-adverse)
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:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Necropsy findings were limited to skin flaking and scabbing first identified in life and observations related to wearing the Elizabethan collar (local alopecia, chromorhinorrhea, and neck lesions) (non-adverse)
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Maternal developmental toxicity

Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
effects observed, non-treatment-related
Description (incidence and severity):
One dam of the mid-dose group (1/25) having a litter consisting of seven early resorptions was pointed out as single non-dosage dependent event and to be within the ranges observed historically at the test facility (non-adverse)
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
not examined
Details on maternal toxic effects:
Maternal toxic effects: yes (local irritation)

Details on maternal toxic effects:
The two highest dose levels caused some local irritation at the site of application, but no decreases in maternal weight gain or feed consumption. Two animals in the control group and one animal in the high-dose group died within 6 h after first application; these were not considered to be treatment related and the animals were replaced. One dam of the mid-dose group (1/25) having a litter consisting of seven early resorptions was pointed out as single non-dosage dependent event and to be within the ranges observed historically at the test facility.
Necropsy findings were limited to skin flaking and scabbing first identified in life and observations related to wearing the Elizabethan collar (local alopecia, chromorhinorrhea, and neck lesions).

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOAEL
Effect level:
2 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Dose descriptor:
NOAEL
Remarks:
local
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
2 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity

Results (fetuses)

Fetal body weight changes:
no effects observed
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
Visceral malformations:
no effects observed
Description (incidence and severity):
An increased number of fetuses with levocardia was observed after in utero exposure to TMP ester of C8/C10 fatty acids (ESIS: Decanoic acid, ester with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol octanoate) (CAS 11138-60-6) via the dermal route. However, as no malformations in the heart were detected and levocardia was also observed in control fetuses of the respective test facility and in a further study (Smith et al. 1988), the toxicological relevance of this effect remains questionable. Thus, incidental occurrence of levocardia cannot be excluded.
Other effects:
not examined
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects: no effects
There were no significant differences from control in any of the developmental parameters measured, including embryo/fetal viability, fetal weight, malformations, or variations. The observed effects in fetuses were not dose-independent and regarded to be sporadic.

Effect levels (fetuses)

Remarks on result:
other: no effects determined

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Any other information on results incl. tables

Table 1: Skin reaction observations

 

0 mg/kg bw/d

200 mg/kg bw/d

600 mg/kg bw/d

2000 mg/kg bw/d

Maximum possible incidencesa

375/25

375/25

375/25

375/25

Erythema

Total

0/0

2/1

22/4

91/13b

Grade 1

0/0

2/1

10/4

81/13b

Grade 2

0/0

0/0

4/1

10/4b

Flaking

Total

11/3

15/2

55/6

170/17 b

Grade 1

11/3

9/2

27/5

61/14 b

Grade 2

0/0

6/1

19/4

71/14b

Grade 3

0/0

0/0

9/1

38/7 b

Edema

Total

0/0

0/0

23/4

83/11b

Grade 1

0/0

0/0

18/4

59/11b

Grade 2

0/0

0/0

5/1

24/6b

Scab

0/0

0/0

6/2

19/4

a:       Maximum incidence : Days x rats from first treatment on GD 6 through sacrifice on GD 20 divided by the number of rats examined per group on GD 6-20

b:        Significantly different from vehicle control group value (p≤0.01)

 

Table 2: Maternal reproductive, litter, and fetal alteration observations: Caesarian-Section results on GD 20

 

0 mg/kg bw/d

200 mg/kg bw/d

600 mg/kg bw/d

2000 mg/kg bw/d

Rats pregnant and sectioned on Day 20 of gestation (n)

25

23

22b

24

Corpora lutea/dam

16.4

16.6

16.9

16.5

Implantation sites/litter

15.0

15.4

14.9

14.2

Litter size

Live fetuses/litter

14.6

14.6

14.0

13.3

Live fetuses (n)

364

335

308

320

Dead fetuses (n)

0

0

0

0

Resorptions

0.4

0.9

0.9

0.9

Early (n)

10

20

19

21

Late (n)

1

0

0

0

Dams with any resorptions n(%)

9 (36)

11 (48)

15 (68)

11 (46)

% resorbed/litter

2.9

5.4

5.8

5.0

% male/litter

51.3

50.8

48.1

47.7

Live fetal body weight (g/litter)

3.68

3.62

3.69

3.75

Male

3.77

3.68

3.82

3.85

Female

3.58

3.56

3.58

3.65

Fetuses evaluated (n)

364

335

308

320

Litters with any alterations observed n(%)

10 (40)

8 (35)

14 (64)

7 (25)

Fetuses with any alterations observed n(%)

13 (3.5)

10 (3.0)

20 (6.5)

9 (2.0)

% fetuses/litter with any alterations observed

3.5

2.9

6.8c

2.7

b:       Excludes values for one dam, which had a litter consisting of seven early resorptions.

c:       Significantly different from vehicle control group value (p≤0.05)

Table 3: Fetal evaluations

 

0 mg/kg bw/d

200 mg/kg bw/d

600 mg/kg bw/d

2000 mg/kg bw/d

Litters evaluated

25

23

22b

24

Fetuses evaluated

364

335

308

320

Live

364

335

308

320

Fetal gross external alterations

364

335

308

320

Tail: kinked

Litter incidence, n (%)

0(0)

1 (4.3)

0(0)

0(0)

Fetal incidence, n (%)

0(0)

1(0.3)

0(0)

0(0)

Body: hematoma

Litter incidence, n (%)

1(4.0)

0(0)

0(0)

0(0)

Fetal incidence, n (%)

1 (0.3)

0(0)

0(0)

0(0)

Fetal soft tissue alterations, evaluations

174

162

149

155

Vessels: umbilical artery descended to the left of urinary bladder

Litter incidence, n (%)

2(8.0)

3(13.0)

2(9.1)

2(8.3)

Fetal incidence, n (%)

2(1.1)

3(1.8)

3(2.0)

2(1.3)

Vessels: apparent additional umbilical artery descended left of the bladder

Litter incidence, n (%)

0(0)

0(0)

1(4.5)

0(0)

Fetal incidence, n (%)

0(0)

0(0)

1(0.7)

0(0)

Fetal skeletal alterations, evaluations

190

173

159

165

Cervical vertebrae: cervical rib present at 7th cervical vertebrae

Litter incidence, n (%)

2(8.0)

1(4.3)

1(4.8)

0(0)

Fetal incidence, n (%)

2(1.0)

2(1.2)

1(1.2)

0(0)

Thoracic vertebrae: centrum, bifid

Litter incidence, n (%)

1(4.0)

1(4.3)

5(22.7)

0(0)

Fetal incidence, n (%)

1(0.5)

1(0.6)

5(3.1)a

0(0)

Lumbar vertebrae: centrum, bifid

Litter incidence, n (%)

0(0)

1(4.3)

0(0)

0(0)

Fetal incidence, n (%)

0(0)

1(0.6)

0(0)

0(0)

Ribs: wavy

Litter incidence, n (%)

0(0)

0(0)

2(9.1)

1(4.2)

Fetal incidence, n (%)

0(0)

0(0)

2(1.2)

1(0.5

Sternal centra: 1st, not ossified

Litter incidence, n (%)

1(4.0)

0(0)

0(0)

2(8.3)

Fetal incidence, n (%)

1(0.5)

0(0)

0(0)

2(1.3)

Sternal centra: 1st, incompletely ossified

Litter incidence, n (%)

3(12.0)

3(13.0)

2(5.1)

1(4.2)

Fetal incidence, n (%)

4(2.1)

4(2.3)

2(1.2)

1(0.6)

Pelvis: pubis, incompletely ossified

Litter incidence, n (%)

3(12.0)

0(0)

4(18.2)

3(12.5)

Fetal incidence, n (%)

3(1.6)

0(0)

5(3.1)

3(1.8)

Pelvis: ischium, incompletely ossified

Litter incidence, n (%)

0(0)

0(0)

2(9.1)

0(0)

Fetal incidence, n (%)

0(0)

0(0)

2(1.2)

0(0)

a: Significantly different from vehicle control group (p≤0.01)

Applicant's summary and conclusion

Conclusions:
In a developmental toxicity study, pregnant rats were dermally exposed to the test substance. No adverse effects were observed in any maternal or reproductive parameter or on external and skeletal development of fetuses. Thus, the developmental NOAEL (maternal/developmental) was determined to be 2000 mg/kg bw. The local NOAEL was considered to be 200 mg/kg bw/day due to local dermal irritation.

Categories Display