Hexanoic acid, 2-ethyl-, C16-18-alkyl esters

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Effect on fertility: via oral route

Endpoint conclusion:

no study available

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

A substance-tailored exposure-driven testing was followed for the hazard assessment of toxicity to reproduction. No significant exposure is expected throughout all relevant exposure scenarios according to Annex XI, section 3.2(a) (i), therefore testing for toxicity to reproduction is omitted in accordance with Annex VIII column 2 section 8.6.1. The justification is based on an exposure assessment in accordance with section 5 of Annex I.

Further details are given in the endpoint itself , as well as in the chapter "Toxicological information" and in chapter 9 and 10 of the CSR.

Effects on developmental toxicity

Description of key information

Prenatal developmental toxicity study (OECD 414), rat, oral:
NOAEL(maternal toxicity): ≥ 1500 mg/kg bw/day
NOAEL(teratogenicity): ≥ 1500 mg/kg bw/day
NOAEL(developmental toxicity): ≥ 1500 mg/kg bw/day

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 May - 13 Jun 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-guideline study with acceptable restrictions. The test substance was only administered during organogenesis (Day 6-15 of gestation).

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

adopted in 2001

Deviations:

yes

Remarks:

Exposure was only from day 6-15 of gestation

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Wistar rats (Chbb: THOM (SPF))

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr. K. THOMAE GmbH, Biberach an der Riss, Germany
- Age at study initiation: 10-11 weeks
- Weight at study initiation: mean approx. 233 g
- Fasting period before study: 5 days
- Housing: the animals were housed singly from Day 0 – 20 p.c. in type DK III stainless steel wire mesh cages supplied by BECKER & CO., Castrop-Rauxel, Germany
- Diet: ground Kliba 343 feed rat/mouse/hamster supplied by KLINGENTALMUHLE AG, Kaiseraugst, Switzerland, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 13 May 1996 To: 13 June 1996

Route of administration:

oral: gavage

Vehicle:

other: olive oil DAB 10

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The dosing solutions were prepared daily before administration, adjusted to the body weight measured on Day 6 of gestation. For the preparation of the solutions, an appropriate amount of the respective test substance was weighed in a volumetric flask, subsequently topped up with olive oil DAB 10 and intensively shaken.

VEHICLE
- Concentration in vehicle: 12 000, 20 000 and 30 000 mg/100 mL
- Amount of vehicle (if gavage): 5 mL/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analytical verifications of the stability of the test substance in olive oil (DAB 10) for a period of 4 hours, at room temperature, was determined before the beginning of this study. Samples of test substance solutions were analysed (by gas chromatography) twice during the study period to verify the concentrations. The analytical values of the samples corresponded to the expected values within the limits of the analytical method. The maximum deviation was about 5% from the target value.

Details on mating procedure:

- Impregnation procedure: cohoused
- M/F ratio per cage: 1-4 female rats were mated with one male
- Length of cohabitation: mating took place from about 4 pm to 7.30 am on the following day.
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

Day 6-15 of gestation

Frequency of treatment:

daily, 7 days/week

Duration of test:

10 days; Day 6-15 of gestation

No. of animals per sex per dose:

25 females per dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: from several prenatal toxicity studies in rats (e.g. PENNANEN et al., 1993,) it is know that 2-ethylhexanoic acid induces overt maternal toxicity at doses of 50—600 mg/kg bw/day (e.g. impairments in maternal body weight gain, increased liver weight) and that 1000 mg/kg bw/day are even lethal for most of the dams. Clear signs of developmental toxicity in the form of reduced fetal body weights and delays in skeletal maturation appeared at doses between 250 and 300 mg/kg bw/day, whereas teratogenic effects were still described by PENNANEN et al. (1993) at 300 mg/kg bw/day and above. Therefore, 600 mg 2-ethylhexanoic acid/kg bw/day were administered to the positive control group in the present study with the aim of inducing clear maternal and developmental toxicity (including signs of teratogenicity), but no excessive maternal toxicity. Taking into account the above, the following doses were chosen for cetylstearyl 2-ethylhexanoate in the main study: 600 mg/kg bw/day – as the expected NOAEL, 1000 mg/kg bw/day – as the intermediate dose level and 1500 mg/kg bw/day – at this dose about 600 mg 2-ethylhexanoic acid/kg bw/day may be released theoretically at this dose.

- Treatment groups:
group 0: 0 mg/kg bw/day - [Vehicle control]
group 1: 600 mg/kg bw/day - [Positive control, 2-ethylhexanoic acid]
group 2: 600 mg/kg bw/day - Cetylstearyl 2-ethylhexanoate
group 3: 1000 mg/kg bw/day - Cetylstearyl 2-ethylhexanoate
group 4: 1500 mg/kg bw/day - Cetylstearyl 2-ethylhexanoate

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily on working days or once a day (Saturday, Sunday or on public holiday)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least once a day

BODY WEIGHT: Yes
- Time schedule for examinations: on Day 0 (prior to administration), 1, 3, 6, 8, 10, 13, 15, 17 and 20 of gestation.

FOOD CONSUMPTION : with the exception of Day 0, 1,3,6,8,10,13,15,17 and 20 of gestation

WATER CONSUMPTION : No

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day 20
- Organs examined: gross macroscopic examination of all reproductive and gender-specific organs, with emphasis on the uterus, uterine contents, ovaries, position of the fetuses in the uterus and number of corpora lutea was performed and the data recorded. Liver and kidney weights were recorded. Weight of the unopeaned uterus was also noted.

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
- Other:
- Number and distribution of implantation sites classified as: live foetuses and dead implantations ( 1 – early resorptions, 2 – late resorptions and 3 – dead foetuses)
- calculations of conception rate and pre- and postimplantation loss were carried out.

Fetal examinations:

- External examinations: Yes, all fetuses. The viability of the fetuses and the conditions of the placentae, the umbilical cords, the fetal membranes and fluids were examined. Individual placental weights were recorded.
- Soft tissue examinations: Yes: [half per litter]. Malformation (e.g. exencephaly, atresia ani, and hernia umbilicalis), variations (e.g. dilated renal pelvis) and unclassified observations (e.g. blood coagulum around placenta).
- Skeletal examinations: Yes: [ half per litter ]. Malformation (e.g. exencephaly, atresia ani, and hernia umbilicalis), variations (e.g. dilated renal pelvis) and retardations (e.g. sternebra (e) not ossified).
- Head examinations: Yes
Live fetuses were sexed, weighed individually including placentae, examined for gross external abnormalities and allocated to one of the following procedures:
1) Half of the fetuses from each litter were non-individually fixed in Bouin's solution in order to examine for any visceral findings according to the method of BARROW and TAYLOR (Barrow and Taylor, 1969). After this examination the foetuses were discarded.
2) The remaining fetuses were placed non-individually in a solution of ethyl alcohol. The skeletons of the fetuses were then stained according to a modified method of DAWSON (Dawson, 1926). Thereafter, the skeleton of these fetuses were examined under a stereomicroscope. After this examination the stained fetal skeletons were retained by litter.

Statistics:

The following statistical methods were used to analyse food consumption, body weights and reproduction data:
• Means and standard deviations of various data were calculated.
• Dunnett-test based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex. Parameters: food consumption, body weight, body weight change, corrected body weight gain (net maternal body weight change), carcass weight, weight of unopened uterus, liver and kidney weights, number of corpora lutea, number of implantations, number of resorptions, number of live fetuses, proportions of pre-implantation loss, proportions of post-implantation loss, proportions of resorptions, proportion of live fetuses in each litter, litter mean fetal body weight and litter mean placental weight.
• Wilcoxon-test. Parameters: proportions of fetuses with malformations, variations, retardations and/or unclassified observations in each litter.
• Fisher's exact-test.Parameters: female mortality, females pregnant at terminal sacrifice and number of litters with fetal findings.

Historical control data:

Historical control data was provided

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
Mortality:
No death occurred in the dams of test group 0 (vehicle control) and in the test groups 2 - 4. Furthermore, no mortalities occurred in test group 1 (positive control).

Signs and/or symptoms:
No abnormal clinical findings were observed in any of the treatment groups or in test group 1 (positive control).

Food consumption:
The food intake of the dams of test groups 2 - 4 was not influenced by the administration of the test substance in comparison with the vehicle control group. There was a slight, sporadically statistically significant increase in food intake in the low and the high dose groups, which was considered to be spontaneous in nature. In the positive control group (test group 1), a marginal, statistically significant reduction in the food consumption was observed at the beginning of the treatment period (Days 6 – 8 gestation).

Body weight and body weight gains:
The mean body weights were similar in all groups and hence, body weight gain was not influenced by test substance administration. However, body weight gain of the dams of test group 3 (1000 mg/kg bw/day) was increased compared to control animals at the beginning of the study period (GD 0 and 1) which is considered as a spontaneous effect without biological significance as no adverse effects were observed in dams or fetuses of this dose group.
The mean bodyweight gains of the dams of the positive control group (test group 1) were statistically significant impaired between days 15-17 gestation.

Corrected body weight gain (net maternal body weight change):
The corrected body weight gains (terminal body weight on Day 20 gestation minus weight of the unopened uterus minus body weight on Day 6 gestation) were comparable among the groups, including test group 1 (positive control).

Organ weights:
There were no substance-related and/or biologically relevant differences in weight of the following organs: uterus, liver and kidneys (absolute and relative) in test substance exposed and control animals. In contrast, the mean relative liver weight of the positive control group (test group 1) was statistically increased (about 6% above the negative control group (test group 0) value).

Reproduction data:
There were no substance-related and/or biologically relevant differences between the different tests groups in conception rate, in the mean number of corpora lutea and implantation sites or in the values calculated for the pre- and the post-implantation losses, the number of resorptions and viable fetuses.
The differences evident (see table 2 for detail) were considered to be incidental and within the normal range of deviations for animals of this strain and age. There was one fetus in the test group 0 (vehicle control) and one in the test group 4 (1500 mg/kg bw/d), which were found dead when the uterus of the respective dams were opened.

Necropsy findings:
No substance-related macroscopic changes were noted in the dams of test group 0 (vehicle control) nor in test groups 2 – 4. Similarly, no substance-related abnormalities were observed in dams of the positive control group. Individual animals revealed edema/emphysema of the lungs (linked to the method of sacrifice), bilateral hydrometra and keratinous cyst of the forestomach. Due to the incidental occurence, these effects are not considered as treatment-related.

Dose descriptor:

NOAEL

Effect level:

>= 1 500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEL

Effect level:

>= 1 500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Body weight:
The weights of live fetuses exhibited no significant differences on a litter and individual basis between the test group 0 (vehicle control) and the test groups 2 – 4. The mean fetal body weights in test group 1 (positive control) were statistically significantly lower than in the test group 0 (negative control), which is considered to be treatment-related and in line with the reductions in mean placental weights in this group.


Placenta and uterus weight:
The weights of placentae and the whole uterus showed no significant differences between the test group 0 (vehicle control) and the test groups 2 – 4. The marginal, but statistically significant, lower mean placental weights at 600 (test group 3) and 1500 (test group 4) mg/kg bw/day were not considered to be of any biological relevance, as these values are within the historical control range. The mean placental weights in test group 1 (positive control) were statistically significantly lower than in the negative control group (test group 0), which was considered to be treatment-related and in line with the reductions in mean fetal body weights in this group.

Sex ratios:
The sex ratio of the fetuses was not affected by the treatment with the test substance or positive control.

External examinations:

No external malformations occurred at 1500 (test group 4) mg/kg bw/day, but anophthalmia was recorded in one low dose fetus and macroglossia in one mid dose fetus. Due to the isolated and disparate nature of these findings and the occurrence of these external malformations in the historical control data at a low rate, a substance induced origin was excluded. The external examination of the fetuses revealed no external variations in any test groups. So-called unclassified observations (blood coagulum around placenta or placentae fused) were recorded for two test group 0 (vehicle control) fetuses and one fetus of the test group 4. The former mentioned fetus in group 0 and the fetus in test group 4 were already dead, when the dams’ uterus was opened. The isolated occurrence of these findings does not suggest any treatment relationship. In test group 1 (positive control), 5 fetuses (1.8 % of the examined fetuses) from 3 litters (14% of the examined litters) showed external malformations (1.7% affected fetuses/litter): - adactyly on both forelimbs: male fetus No.2 of dam No.29 and - different tail malformations (shortened, filiformed or no tail [acaudia]): fetuses No.2 (male) of dam No.29, No. 9 (male) of dam No.29, No.9 (male) of Dam No.29, No. 11 (male) of dam No.35, Nos. 9 (male), and 17 (female) of dam 40.


Soft tissue examination:

Soft tissue malformation: One animal in the test group 0 (vehicle control) had malposition of the heart (dextrocardia). In test groups 2 (600 mg/kg bw/day) one male fetus had globular shaped heart (both ventricles dilated). Furthermore, one female fetus with transposition of great vessels was observed. The mean percentage of affected fetuses/litter (1.1%) is within the respective historical control range of 0 – 1.9 %, and because there is no dose-response relationship, a substance induced orgin was excluded. No soft tissue malformations were observed in any fetuses in test groups 3 and 4 (1000 or 1500 mg/kg bw/day). However, in test group 1 (positive control) one male fetus with displacement of the aortic arch (malformation of great vessels) and one female fetus with globular shaped heart (both ventricles dilated) were observed, the mean percentage of affected fetuses/litter (5.3%) is above the respective historical control range [0.4 % (0-1.9%)].
Soft tissue variations: Two fetal soft tissue variations (dilated renal pelvis and hydroureter) were detected in all groups including test group 0 (vehicle control). The fetal/litter incidences and the mean percentages of affected fetuses/litter, in the rat strain used, are within the range of biological variation and do not show any relation to dosing.
Unclassified observations: No so-called unclassified observations (e.g. bloody imbibition of kidney(s)) were recorded.

Skeletal examination:

Skeletal malformation: One animal in the test group 0 (vehicle control) showed two skeletal malformations in the form of absent lumber vertebra and bifurcated rib(s) [0.7% of the examined fetuses] from one litter [4.5% of the examined litters], 1.5% affected fetuses/litter.
In test groups 2 – 4 (600, 1000 and 1500 mg/kg bw/day) the rate of skeletal malformations did not show a clear dose-response relationship. The malformations were related to the vertebral column (thoracic and/or lumber vertebra absent, fused and/or of irregular shape), the sternum (sternebra (e) bipartite, ossification centres dislocated) and the ribs (fused). The total rate of fetuses with skeletal malformations was statistically significantly elevated against test group 0 (vehicle control) at 1000 mg/kg bw/day (particularly bipartite sternebra (e) with dislocated ossification centres), but not in the other test groups. Most of the observed skeletal malformations or very similar ones which are found in a comparable frequency in the historical control data and are therefore considered to be without any biological relevance.

Skeletal variations: The skeletal variations elicited in test group 0-4 include:
- Ribs (wavy, flying, shortended 13th, accessory 14th or rudimentary cervical rib(s),
- The sternum (accessory sternebra, sternebra(e) of irregular shape or bipartite,
- The skull (epactal bone between parietal and intraparietal or frontal and parietal bones),
- The vertebral column (accessory thoracic or lumber vertebra) and
- The clavicular (deformed).
While skeletal variations occurred at similar incidences in the test group 0 (vehicle control) and in test groups 2 – 4 (600, 1000 and 1500 mg/kg bw/day), they were found at statistically significantly increased rates in the positive control group (test group 1).


Skeletal retardations: The statistically significant increased occurrence of just one skeletal retardation (incomplete ossification of thoracic vertebral body/bodies) in fetuses of test groups 2 and 4 ( 600 or 1500 mg/kg bw/d) was considered to be spontaneous in nature and without biological relevance, because no clear dose-response is shown and the rates for this particular finding (9.1 or 9.2 % affected fetuses/litter, respectivey) is within the historical control range [8.1% (0 – 50%)]. In the positive control group (test group 1), skeletal retardations (like delays in general ossification and ossification of the extremities and the pelvic girdle) were found at statistically significant increased rates. All except one positive control fetuse showed delays in skeletal maturation, which is in line with the reduced fetal body weight in this test group.

Positive control group (test group 1) – The overall rate of malformed fetuses/litter (7.1%) was statistically significantly increased, particularly the occurrence of rare limb (adactyly), tail (shortened, filiformed or absent tail [acaudia]) and vertebral column malformations as these incidences are outside the historical control data. Furthermore, fetal and litter incidences for skeletal and overall variations (especially accessory vertebra; rudimentary cervical, accessory 14th and wavy rib(s)) and skeletal and overall retardation and the mean percentage of fetuses/litters with these findings were distinctly increased and are above the historical control range.

Dose descriptor:

NOAEL

Effect level:

>= 1 500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Abnormalities:

not specified

Developmental effects observed:

not specified

Table 2. Summary of reproduction data

 

Parameters		Group 0 -   0 mg/kg bw/d	Group 1 – 600 mg/kg bw/d (2-EHA)	Group 2 - 600 mg/kg bw/d (LVT)	Group 3 - 1000 mg/kg bw/d(LVT)	Group 4 - 1500 mg/kg bw/d (LVT)
Females mated	N	25	25	25	25	25
Pregnant	N	22	22	25	22	24
Conception Rate	%	88	88	100	88	96
Aborted	N	0	0	0	0	0
Premature births	N	0	0	0	0	0
Dams with viable fetuses	N	22	22	25	22	24
Dams with all resorptions	N	0	0	0	0	0
Female Mortality	N	0^F	0^F	0	0	0
	%	0.0	0.0	0.0	0.0	0.0
Pregnant at terminal sacrifice	N	22^F	22^F	25	22	24
	%	88	88	100	88	96
Corpora lutea	Mean	15.4^D	15.7^D	16.0	15.7	15.4
	S.D.	1.65	1.88	1.84	2.32	1.28
	Total	339	346	401	345	370
Implantation sites	Mean	13.4^D	14.1^D	15.0	14.2	14.4
	S.D.	3.96	3.52	2.32	3.46	2.36
	Total	295	310	376	313	345
Preimplantation loss	Mean	13.6^D	11.0^D	6.2	9.4	6.7
	S.D.	23.45	18.66	10.04	17.75	13.26
Post-implantation loss	Mean	8.0^D	10.4^D	8.7	11.1	8.5
	S.D.	8.33	9.43	9.47	8.65	6.43
Resorption: total	Mean	1.0^D	1.4^D	1.3	1.6	1.2
	S.D.	0.95	1.33	1.38	1.6	1.2
	Total	21	31	33	36	28
	Mean %	7.7^D	10.4^D	8.7	11.1	8.2
	S.D.	8.05	9.43	9.47	8.65	6.36
Early	Mean	0.9^D	1.2^D	1.1	1.6	1.0
	S.D.	0.92	1.19	1.29	1.47	0.83
	Total	20	27	27	35	24
	Mean %	7.5^D	9.2^D	7.3	10.3	6.9
	S.D.	7.99	8.73	9.13	8.86	5.79
Late	Mean	0.0^D	0.2^D	0.2	0.0	0.2
	S.D.	0.21	0.39	0.52	0.21	0.38
	Total	1	4	6	1	4
	Mean %	0.3^D	1.1^D	1.4	0.8	1.3
	S.D.	1.33	2.49	3.05	3.55	3.07
Dead fetuses	N	1	0	0	0	1

 

Statistics: D= Dunnett-test (two-sided) F= Fisher’s exact test (one-sided). *: P<=0.05. **:P<=0.01. 2-EHA = 2-ethylhexanoic acid. LVT = cetylstearyl 2-ethylhexanoate

 

Table 3. Mean placental and fetal body weights.

Parameters		Group 0 -  0 mg/kg bw/d	Group 1 – 600 mg/kg bw/d (2-EHA)	Group 2 - 600 mg/kg bw/d (LVT)	Group 3 - 1000 mg/kg bw/d(LVT)	Group 4 - 1500 mg/kg bw/d (LVT)
Placental weights in grams
of all viable fetuses	Mean	0.46^D	0.40^D	0.43*	0.45	0.43*
	S.D.	0.057	0.040	0.039	0.040	0.029
	N	22	22	25	22	24
of male fetuses	Mean	0.47^D	0.40^D	0.43*	0.46	0.44
	S.D.	0.060	0.042	0.038	0.044	0.032
	N	22	22	25	22	24
of female fetuses	Mean	0.46^D	0.40^D	0.43	0.45	0.42*
	S.D.	0.059	0.039	0.039	0.042	0.028
	N	22	22	25	22	24
Fetal weights in grams
of all viable fetuses	Mean	3.8^D	3.0^D	3.7	3.8	3.7
	S.D.	0.20	0.22	0.23	0.20	0.22
	N	22	22	25	22	24
of male fetuses	Mean	3.9^D	3.1^D	3.8	3.8	3.8
	S.D.	0.25	0.27	0.23	.024	0.21
	N	22	22	25	22	24
of female fetuses	Mean	3.7^D	3.0^D	3.6	3.6	3.6
	S.D.	0.18	0.22	0.23	0.19	0.25
	N	22	22	25	22	24

  Statistics: D= Dunnett-test (two-sided)*: P<=0.05. **:P<=0.01. 2-EHA = 2-ethylhexanoic acid. LVT = cetylstearyl 2-ethylhexanoate

Table 4. Summary of all classified fetal external observations

Parameters		Group 0 - 0 mg/kg bw/d	Group 1 – 600 mg/kg bw/d (2-EHA)	Group 2 - 600 mg/kg bw/d (LVT)	Group 3 - 1000 mg/kg bw/d(LVT)	Group 4 - 1500 mg/kg bw/d (LVT)
Litter evaluated	N	22	22	25	22	24
Fetuses evaluated	N	274	279	343	277	317
Live	N	273	279	343	277	316
Dead	N	1	0	0	0	1
Total malformations
Fetal incidence	N	0	5	1	1	0
	%	0.0	1.8	0.3	0.4	0.0
Litter incidence	N	0^F	3	1	1	0
	%	0.0	14	4.0	4.5	0.0
Affected fetuses/litter	Mean %	0.0^W	1.7^A	0.3	0.3	0.0
	S.D.	0.0	4.67	1.33	1.52	0.00
Total variations
Fetal incidence	N	0	0	0	0	0
	%	0.0	0.0	0.0	0.0	0.0
Litter incidence	N	0^F	0	0	0	0
	%	0.0	0.0	0.0	0.0	0.0
Affected fetuses/litter	Mean %	0.0^W	0.0	0.0	0.0	0.0
	S.D.	0.00	0.00	0.00	0.00	0.00

Statistics: F= Fisher’s exact test (one-sided).W = Wilcoxon-test (one-sided). A: p <=0.05 using control #1 B: p <=0.01 using control #1.

2-EHA = 2-ethylhexanoic acid. LVT = cetylstearyl 2-ethylhexanoate

Conclusions:

CLP: not classified
DSD: not classified

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 2), and is thus sufficient to fulfill the standard information requirements set out in Annex VIII-IX, 8.7 of Regulation (EC) 1907/2006.

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

The potential of hexanoic acid, 2-ethyl-, C16-18 alkyl esters (CAS 90411-68-0) to cause developmental toxicity was assessed in a study performed according to OECD guideline 414 and in compliance with GLP (Hellwig, J., 1997). 22 to 25 pregnant Wistar rats were treated by oral gavage with the test substance from gestation day 6 until day 20 at doses of 600, 1000 and 1500 mg/kg bw/day. A concurrent negative control group (vehicle only) and a positive control group (2-ethylhexanoic acid) were included in this study. No mortalities or no compound-related symptoms were observed in dams of all test groups. In addition, body weight and body weight gains were within the expected ranges. No compound-related differences were noted in conception rate, mean number of corpora lutea, implantation sites, pre- and post- implantation losses, the number of resorptions and viable fetuses after administration of the test substance.

At scheduled necropsy no macroscopic changes were noted in the dams of the treatment groups. Furthermore, the mean number of total fetuses was not affected by treatment with the test substance. In addition, mean fetal placental and uterus weights were not affected. The fetal sex ratio was comparable in all groups and no treatment-related fetal abnormalities were found at necropsy. The examined fetuses showed no treatment-related malformations in the test groups compared to the negative control group. The various external, skeletal and soft tissue malformations observed in the study were observed sporadically and lay within the range of historical controls. Therefore, these effects were not considered as substance-related. As described in the available reproductive and postnatal development study (Pennanen et al. 1993), oral administration of 600 mg 2-ethylhexanoic acid/ kg bw/day as a ‘positive control’ revealed some signs of maternal toxicity (reduction in food consumption, impairment in body weight gains and increased liver weights) and clear indications for selective developmental toxicity (external malformations, skeletal variations and skeletal retardations), including indications for teratogenicity (impairment in body weight gains and lower mean placental weights). In the positive control group, the overall rate of malformed fetuses/litter, fetal and litter incidences for skeletal and overall variations (especially accessory vertebra; rudimentary cervical, accessory 14th and wavy rib(s)) and skeletal and overall retardation and the mean percentage of fetuses/litters with these findings were distinctly increased and were above the historical control range. Based on the lack of adverse effects in this study, the NOAEL for maternal and developmental toxicity for rats was considered to be above 1500 mg/kg bw/day.

Justification for selection of Effect on developmental toxicity: via oral route:
There is only one study available.