Amines, C11-C13 (linear and branched) alkyl

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Additional information

There is no specific fertility study with TDA available. TDA was tested under GLP conditions in a 90 day oral gavage study according to OECD TG 408 in male and female Wistar rats (BASF AG, 2004; Val. 2). The tested dose levels were 5, 15, and 45 mg/kg bw/day. Additionally to the requirements of OECD TG 408, the effects on the male and female reproductive organs were examined in this study in terms of absolute and relative weight changes, histopathological, and functional changes of the male and female reproductive organs. Sperm parameters were determined on all animals with respect to sperm motility, sperm morphology, sperm head count (cauda epididymidis and testis). Estrous cycle was determined in all females from day 63 to 91.

 

No weight changes were seen in the reproductive organs apart from a significantly decreased absolute weight of the testes. As this effect was only found in the mid-dose group, and not in the high-dose animals, it was considered to be not related to the administration of the test substance. This also holds true for the prolonged estrous cycle of the mid-dose females. As there was no dose relation observed, this finding was also considered not to be related to the treatment with Tridecylamine. Sperm parameters were unchanged compared to controls. Thus, the NOAEL for the examined fertility parameters in males and females was 45 mg/kg bw/day, the highest dose tested.

Effects on developmental toxicity

Description of key information

NOAEL: maternal toxicity: 20 mg/kg bw/day (OECD TG 414)
NOAEL: teratogenicity: >= 80 mg/kg bw/day (OECD TG 414)

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes (incl. QA statement)

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Germany
- Age at study initiation: 70 - 84 days
- Weight at study initiation: 146.4 - 183.2 g
- Housing: single housing from day 0- 20 post coitum in type DK III stainless steel wire mesh cages (BECKER & CO., Castrop-Rauxel, FRG; height: 15 cm x 37.5 cm x 21 cm; floor area about 800 cm²).
- Diet: ground Kliba maintenance diet rat/mouse/hamster meal (PROVIMI KLIBA SA, Kaiseraugst, Switzerland) ad libitum
- Water: Tap water ad libitum
- Acclimation period: 6 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12 / 12 (6.00 pm - 6.00 am dark; 6.00 am - 6.00 pm light)

Route of administration:

oral: gavage

Vehicle:

other: 0.5% Carboxymethylcellulose in doubly distilled water with 0.05% Cremophor EL

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The aqueous test substance emulsions were prepared at the beginning of the administration period and thereafter at intervals which took into account the analytical results of the stability verification. For the preparation of the emulsions, an appropriate amount of the test substance was weighed depending on the dose group in calibrated beakers, topped up with 0 .5% Carboxymethylcellulose in doubly distilled water with 0.05% Cremophor EL and subsequently thoroughly mixed using a magnetic stirrer. This devise was also used to keep the emulsions homogeneous during treatment of the
animals.


ADMINISTRATION VOLUME:
10 ml/kg bw/d

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The test substance emulsions were analyzed by GC. Samples of the test substance emulsions were sent to the analytical laboratory twice during the study period (at the beginning and towards the end) for verification of the concentrations. The samples which were taken for the first concentration control analyses at the beginning of the administration period were also used to verify the homogeneity for the samples of the low and the high concentrations (5 and 80 mg/kg body weight/day). 3 samples (one from the top, middle and bottom in each case) were taken for each of these concentrations from the beaker with a magnetic stirrer running.

Details on mating procedure:

The animals were mated by the breeder ("time-mated") and supplied on day 0 post coitum (p.c. = detection of vaginal plug / sperm).

Duration of treatment / exposure:

gestation days 6-19

Frequency of treatment:

daily

Duration of test:

3 weeks

Remarks:

Doses / Concentrations:
0, 5, 20, 80 mg/kg bw/d
Basis:

No. of animals per sex per dose:

24 females

Control animals:

yes, concurrent vehicle

Maternal examinations:

The animals were examined at least daily for clinical symptoms. Mortality was checked twice a day or once on Saturday/Sunday. Food consumption and body weight on gestation days 0, 1, 3, 6, 8, 10, 13, 15, 17, 19 and 20 were recorded. Corrected body weight gain was calculated after terminal sacrifice as [bw on gestation day 20 - (uterus weight+bw gestation day 6)]. Dams were sacrificed on gestation day 20. Necropsy included gross pathology assessment. Uterus and ovaries were removed with subsequent examination.

Ovaries and uterine content:

Examination and recording of: unopened uterus weight; no. of corpora lutea; no. and classification of implantation sites as live fetuses or dead implantations (early or late resorptions, dead fetuses). Conception rate and pre- and postimplantation losses were calculated.

Fetal examinations:

Fetuses were weighed and sexed by anogenital distance observation (which was later confirmed by internal examination of those fetuses which were preserved in BOUIN's solution), and macroscopically examined (viability, condition of placenta, fetal membranes, umbilical cords, placental weight). Approx. 50% of all fetuses were subjected to soft tissue examinations after fixation in BOUIN's solution, the other 50% of fetuses was examined for skeletal changes.

Statistics:

Statistical evaluation of data included FISHER's Exact Test for conception rate, mortality of the dams, and number of litters with fetal findings; WILCOXON Test for proportions of fetuses with malformations and/or variations in each litter; and DUNNETT's Test for all other data including water consumption and body weight gain.

Details on maternal toxic effects:

Details on maternal toxic effects:
Only pregnant dams or pregnant dams sacrificed at schedule were used for calculations. Animals which were totally or partially excluded were 2 non-pregnant animals in each of the groups except the high dose group.
Mortality: none in any of the groups.
Clinical data and signs:
Low dose, 5 mg/kg bw/d: no substance related effects seen.
Intermediate dose, 20 mg/kg bw/d: no substance related effects seen, except transient salivation in two dams during gestation days 8-19. Salivation started immediately after dosing and persisted for some minutes.
High dose, 80 mg/kg bw/d: Transient salivation was noted in 24/24 animals during gestation days 6-19 starting shortly after dosing and lasting few minutes. Additionally, urine smeared fur (4/24 animals) and piloerection (1/24) was noted. Salivation was regarded to be treatment-related due to bad taste or local affection of the upper digestive tract, but was not assessed as an adverse or toxic effect. Food intake was significantly reduced on most of the days. If calculated for the entire treatment period it was 8% less than in the concurrent control. Statistically significant effects on body weight parameters were noted: lowered mean body weight on gestation day 13 (-5%) and lowered corrected body weight gain (-21%) whereas body weight was comparable in all groups throughout the treatment period.
Terminal examination of dams: The mean gravid uterus weight was not influenced by the administration of the test substance. There were no substance-related findings at necropsy in any of the dams. Conception rates reached 92% in control, low, and intermediate dose groups; it was 100% in the high dose group. However, there were no substance-related and/or biologically relevant differences between the test groups in conception rate, mean number of corpora lutea and implantation sites or in the values calculated for the pre- and postimplantation losses, the number of resorptions and viable fetuses.

Dose descriptor:

NOAEL

Effect level:

20 mg/kg bw/day

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

Details on embryotoxic / teratogenic effects:

Details on embryotoxic / teratogenic effects:
Prenatal developmental toxicity (delayed ossification) only at maternally toxic doses. No signs of teratogenicity up to and including the highest dose tested.
Examination of fetuses: Sex distribution was unchanged in all treatment groups. In high dose animals, placental weight was statistically significantly increased (+12% vs. concurrent control), but well in the historical control range. Therefore this was considered to be spontaneous in nature and without toxicological relevance. Mean fetal body weight was unchanged.
The isolated and scattered occurrence of external malformations (anophthalmia, absent tail, cleft palate) in all animal groups including controls did not attain statistical significance in any of the test groups and did not suggest any relationship to treatment. No external variations were noted.
No soft tissue malformations nor unclassified soft tissue observations were seen. The noted soft tissue variations noted (uni- or bilateral dilations of the renal pelvis) were found in fetuses from all test groups without significant differences between the test groups.
Skeletal malformations were noted in single fetuses from all test groups in proportions of 4/111 fetuses (3.6%; control group), 2/108 fetuses (1.9%), 2/109 fetuses (1.8%), and 4/116 fetuses (3.4%) in the low, mid, and high dose group, respectively. No relation to dosing was seen, and no difference to historical control data was noted.
Statistically significantly increased incidences of some skeletal variations were seen in the high dose group fetuses as follows: general delays in skeletal ossification (9.2% fetuses/litter vs. 0% in controls); incomplete ossification of supraoccipital (28.6% fetuses/litter vs. 16.5% in controls); bipartite ossification of thoracic centrum (4.2% fetuses/litter vs. 0% in controls); incomplete ossification of sacral arch (57.4% fetuses/litter vs. 25.7% in controls). The overall incidence of skeletal variations was not different from historical control data.
Thus, there are slight indications of substance-induced effects on skeletal maturation without effects on fetus weight.

Dose descriptor:

NOAEL

Effect level:

>= 80 mg/kg bw/day

Remarks on result:

not determinable due to absence of adverse toxic effects

Abnormalities:

no effects observed

Developmental effects observed:

no

Individual external malformations in fetuses:

Test group (mg/kg bw/d)	Number and sex of fetuses	Malformation	Incidence (%) on the basis of
			fetuses	litters
0 (Control)	1, f	Acaudate (+)	0.5	4.5
5	1, m	Anophthalmia	0.5	4.5
20	-		0	0
80	2, m	Cleft palate	0.9	8.3

f = female; m = male; (+) = fetus with associated skeletal malformations

.

Individual fetal skeletal malformations:

Test group (mg/kg bw/d)	Number and sex of fetuses	Malformation	Incidence (%) on the basis of
			fetuses	litters
0 (Control)	1, f	Misshapen lumbar vertebra, absent sacral vertebra, absent caudal vertebra (+)	3.6	18
	3, m	Misshapen humerus
5	1, f	Misshapen humerus	1.9	9.1
	1, m	Misshapen humerus
20	1, f	Misshapen humerus	1.8	9.1
	1, m	Misshapen humerus
80	1, m	Cleft sternum (changed cartilage), absent tuberositas deltoidea	3.4	17
	3, m	Misshapen humerus

f = female; m = male; (+) = fetus with associated external malformations

.

Occurrence of statistically significantly increased fetal skeletal variations (expressed as mean percentage of affected fetuses/litter):

	Test group (mg/kg bw/d)				HCD Mean % (range)
Finding	0 (Control)	5	20	80
General delays in skeletal ossification	0.0	0.0	0.0	9.2*	Not present
Incomplete ossification of supraoccipital; unchanged cartilage	16.5	24.5	14.0	28.6*	14.0 (5.4 – 21.2)
Bipartite ossification of thoracic centrum; dumbbell-shaped cartilage of centrum	0.0	0.9	0.0	4.2*	1.1 (0.0 – 3.2)
Fused sacral centrum and arch; unchanged cartilage	0.0	5.5**	3.9*	0.0	2.5 (0.0 – 5.9)
Incomplete ossification of sacral arch; cartilage present	25.7	37.4	18.6	57.4**	15.3 (0.0 – 46.1)
Unilateral ossification of sternebra; unchanged cartilage	0.0	3.1*	0.0	3.5*	1.5 (0.0 – 3.6)
Total fetal skeletal variations	98.1	96.5	89.5	95.6	94.1 (87.0 – 99.2)

* = p ≤ 0.05, ** = p ≤ 0.01; HCD = historical control data

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

80 mg/kg bw/day

Additional information

TDA was examined in a GLP developmental/teratogenicity study according to OECD TG 414 using pregnant female Wistar rats (BASF AG, 2003; Val. 1). Groups of 25 animals received dose levels of 0, 5, 20, or 80 mg/kg bw/day during days 6 through 19 of gestation (post coitum). The dams of the high dose group showed signs of toxicity as substantiated by a statistically significant impaired bodyweight gain and a corrected terminal body weight gain lowered by 21 %. No significant clinical sign was noted in the other dose groups.

 

At scheduled necropsy, 22-24 animals/group had implantation sites. The conception rates reached 92 % in all treated groups and were somewhat lower than in the control group (100 %); this small decrease is not considered to be of any biological relevance. There were also no substance-related or biologically relevant differences between the groups with regard to mean number of corpora lutea and implantation sites, pre- and post-implantation losses, or the numbers of resorptions or viable fetuses.

 

The fetal weight and sex distribution was unchanged by the treatment. The occurrence of external malformations, skeletal and soft tissue malformations was comparable between the control and the treated test groups. A statistically significant increased incidence of some skeletal variations was, however, seen in fetuses from the high-dose females. The described findings are considered as skeletal variations because they also occur spontaneously in the strain of rats used and because they reflect a transient effect that is fully reversible postnatally. These were characterized as general delays in skeletal ossification (9.2 % fetuses/litter vs. 0 % in controls); incomplete ossification of supraoccipital (28.6 % fetuses/litter vs. 16.5 % in controls); bipartite ossification of the thoracic centrum (4.2 % fetuses/litter vs. 0 % in controls); incomplete ossification of the sacral arch (57.4 % fetuses/litter vs. 25.7 % in controls). The overall incidence of skeletal variations was not different from historical control data.

 

There were no signs of teratogenicity noted at any dose level. In summary, there were slight indications of substance-induced effects on skeletal maturation without effects on fetal weight at a maternally toxic dose of 80 mg/kg bw/day. Gestational parameters were unaffected up to and including 80 mg/kg bw/day. Fetal parameters were unaffected at all dose levels except the increased occurrence of transient delays in the ossification of the skull and the ventral column in the high dose group. The overall incidence of skeletal variations was not different from historical control data.

 

The NOAEL for maternal toxicity was 20 mg/kg bw/day in this study. There was no sign of teratogenicity at the highest tested dose of 80 mg/kg bw/day.

Justification for classification or non-classification

No need for classification according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.