Isooctadecanoic acid, reaction products with tetraethylenepentamine

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

yes

Remarks:

Deviations are minor and did not negatively impact the quality or integrity of the data nor the outcome of the study.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of fatty acids, C14-C18 (branched and linear) and C18 (unsaturated) with tetraethylenepentamine (linear, branched, cyclic)

EC Number:

701-204-9

Cas Number:

68784-17-8

IUPAC Name:

Reaction products of fatty acids, C14-C18 (branched and linear) and C18 (unsaturated) with tetraethylenepentamine (linear, branched, cyclic)

Test material form:

liquid: viscous

Test animals

Species:

rat

Strain:

other: Crl:CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River
- Age at study initiation: (P) 11 wks
- Weight at study initiation: (P) Males: 355-411 g; Females: 226-276 g
- Housing: F0 animals were housed individually in stainless steel wire-mesh cages suspended above cage-board.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days

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

Administration / exposure

Vehicle:

corn oil

Details on exposure:

The appropriate amount of the test article for each group was weighed into a tared, calibrated glass container. Approximately 70% of the total volume of vehicle was added to each container. The formulations were mixed until uniform using a magnetic stirrer. Vehicle was added to each container to bring the formulations to the calibration mark, and the formulations were stirred until uniform using a magnetic stirrer. The 90 and 200 mg/mL formulations were prepared every 3 days as single formulations for each dosage level, divided into aliquots for daily dispensation and stored at room
temperature. The 30 mg/mL formulations were prepared daily and used within 6 hours of preparation. The test article formulations were stirred continuously throughout the preparation, sampling and dose administration procedures. The test article formulations were visually inspected by the study director on 5 June 2006, and were found to be visibly homogeneous and acceptable for dose administration.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Samples (1 mL each) were collected weekly from the middle stratum of each dosing formulation (including the control group). The samples from the first, fourth and last weeks of dose administration were analyzed for concentration verification; the remaining samples were stored at room temperature for possible future analysis. Additional samples were collected from the 200 mg/mL formulation prepared on 10 July 2006 because the initial analysis revealed a concentration that was higher than expected.

Duration of treatment / exposure:

The males were dosed during study days 0-42 (14 days prior to pairing through 1 day prior to scheduled euthanasia) for a total of 43 doses. The females were dosed during study days 0 through the day prior to euthanasia (14 days prior to pairing through lactation day 3) for a total of 39-43 doses. The female with no evidence of mating was dosed through the day prior to euthanasia (post-cohabitation day 25) for a total of 52 doses.

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

150 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

450 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

12

Control animals:

yes, concurrent vehicle

Examinations

Statistics:

All statistical tests were performed using appropriate computing devices or programs. Analyses were conducted using two-tailed tests (except as noted otherwise) for minimum significance levels of 1% and 5%, comparing each test article-treated group to the control group by sex. Parental mating, fertility, conception and copulation indices were analyzed using the Chi-square test with Yates’ correction factor (Hollander and Wolfe, 1999). Mean parental body weights (weekly, gestation and lactation), body weight changes and food consumption, offspring body weights and body weight changes, gestation length, numbers of implantation sites, numbers of corpora lutea, number of pups born, live litter size on PND 0, unaccounted-for sites, absolute and relative organ weights, and pre-coital intervals values were subjected to a parametric one-way analysis of variance (ANOVA) (Snedecor and Cochran, 1980) to determine intergroup differences. If the ANOVA revealed statistically significant (p<0.05) intergroup variance, Dunnett's test (Dunnett, 1964) was used to compare the test article-treated groups to the control group. Mean litter proportions (percent per litter) of males at birth and postnatal survival were subjected to the Kruskal-Wallis nonparametric ANOVA (Kruskal and Wallis, 1952) to determine intergroup differences. If the ANOVA revealed statistically significant (p<0.05) intergroup variance, Dunn’s test (Dunn, 1964) was used to compare the test article-treated groups to the control group. Histopathological findings in the test article-treated groups were compared to the control group using a two-tailed Fisher’s Exact test (Steel and Torrie, 1980).

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

All males / females survived to the scheduled necropsy. Salivation and evidence thereof (clear
material around the mouth) were observed at a low incidence in 5, 7 and 10 males the
150, 450 and 1000 mg/kg/day groups, respectively, at the time of dose administration;
these findings were first observed on study day 5, study day 6 and study day 2 in these
respective groups. The salivation-related findings were attributed to the test article, but
were not considered adverse because only clear material around the mouth was observed
in single animals in the 450 and 1000 mg/kg/day groups 1-2 hours following dose
administration.
Other clinical findings, including hair loss or scabbing on the forelimbs, yellow material
on the urogenital area, red material around the eyes or nose, lacrimation and/or decreased
defecation, occurred in single animals and/or were not observed in a dose-related manner;
no relationship to the test article was evident.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

MALES : There were no test article-related effects on mean body weights or body weight gains in the 150, 450 and 1000 mg/kg/day group males; none of the differences from the control
group were statistically significant. A slightly lower mean body weight gain was noted in
the 1000 mg/kg/day group males (18 g) during study days 0-7 compared to the control
group (25 g). This reduction was due to 5 males that lost weight or had low body weight
gains. Because the reduction in mean body weight gain in this group during the first
week of treatment was not of sufficient magnitude to result in lower mean body weights throughout the study and the difference from the control group was not statistically
significant, the reduction in mean body weight gain during study days 0-7 was not
considered test article-related.
FEMALES : There were no test article-related effects on mean female body weights or body weight gains during the pre-mating period at any dosage level. Differences from the control
group were slight and not statistically significant.
FEMALES PREGNANCY : There were no test article-related effects on mean maternal body weights or body weight gains during gestation at any dosage level. The only statistically significant (p<0.05) differences from the control group were lower mean body weight gains in the 450 and 1000 mg/kg/day groups (29 g for both groups compared to 36 g in the control group)
during gestation days 14-17. These reductions resulted in slightly lower mean body
weight gains (not statistically significant) when the entire gestation period (gestation days
0-20) was evaluated. Because there was no dose-response relationship, the reductions in
mean body weight gain in the 450 and 1000 mg/kg/day groups were not considered test
article-related.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

MALES : Mean food consumption, evaluated as g/animal/day and g/kg/day, in the 150, 450 and 1000 mg/kg/day group males was similar to that in the control group throughout the
study. The only statistically significant (p<0.05) difference was higher mean food
consumption (evaluated as g/kg/day) in the 450 and 1000 mg/kg/day groups during study
days 35-42.
FEMALES : Mean food consumption, evaluated as g/animal/day and g/kg/day, in the 150, 450 and 1000 mg/kg/day group females was similar to that in the control group during the
pre-mating period. No statistically significant differences were observed.
FEMALES PREGNANCY : Mean maternal food consumption in the 150, 450 and 1000 mg/kg/day groups was
unaffected by test article administration. The only statistically significant (p<0.05)
differences consisted of slightly lower food consumption (2 g/animal/day) in the
150 mg/kg/day group during gestation days 14-17 and slightly higher food consumption
(7 g/kg/day) in the 1000 mg/kg/day group during gestation days 4-7 and 11-14 compared
to the control group.

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

Endocrine 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

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Test article-related histopathologic changes were not observed in the tissues examined
microscopically. All findings were considered to represent common, spontaneous
alterations in laboratory rats.

Histopathological findings: neoplastic:

not examined

Other effects:

effects observed, non-treatment-related

Description (incidence and severity):

One female each in the 450 and 1000 mg/kg/day groups failed to deliver. Both females
were nongravid. Female no. 30050 in the 450 mg/kg/day group also had clear fluid
contents in the uterus, with both horns thickened, and the right uterine horn was attached
to the abdominal wall. These findings were not attributed to the test article.
There were no test article-related macroscopic findings observed at the scheduled
necropsy on lactation day 4. Female no. 30010 in the 1000 mg/kg/day group had white
areas in the lung. No other macroscopic findings were noted.
There were no test article-related effects on the mean number of corpora lutea,
implantation sites or unaccounted-for sites in the 150, 450 and 1000 mg/kg/day groups.
The mean number of implantation sites in the 1000 mg/kg/day group (14.8 per dam) was
lower (not statistically significant) than the concurrent control group value (16.3 per
dam); however, the value in the 1000 mg/kg/day group was within the range of the WIL
historical control data (13.2-17.1 per dam). This reduction was attributed to 1 female that
only had 8 corpora lutea and 4 implantation sites; the mean number of implantation sites
for this group was 15.9 when that female was excluded. Therefore, due to the small
sample size (N=11) and the subsequent greater impact of a single female, the reduction in
the mean number of implantation sites in the 1000 mg/kg/day group was not considered
test article-related. The mean number of implantation sites in the 150 mg/kg/day group
(14.8 per dam) was also lower than the control group value; however, the value in the
450 mg/kg/day group was similar to that in the control group. Therefore, there was no
dose-response relationship in the 150 mg/kg/day group.

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

Test article-related histopathologic changes were not observed in the tissues examined
microscopically. All findings were considered to represent common, spontaneous
alterations in laboratory rats or changes typically associated with pregnancy and lactation

Reproductive function: sperm measures:

no effects observed

Description (incidence and severity):

No test article-related effects on male reproductive performance were noted at any dosage
level. Male mating indices were 100.0%, 100.0%, 91.7% and 100.0% in the control, 150,
450 and 1000 mg/kg/day groups, respectively. Male fertility indices were 100.0%,
100.0%, 91.7% and 91.7% and copulation indices were 100.0%, 100.0%, 100.0% and
91.7% in the same respective groups

Reproductive performance:

no effects observed

Description (incidence and severity):

No test article-related effects on male/female reproductive performance were noted at any dosage
level. Male/female mating indices were 100.0%, 100.0%, 91.7% and 100.0% in the control, 150,
450 and 1000 mg/kg/day groups, respectively. Male fertility indices were 100.0%,
100.0%, 91.7% and 91.7% and copulation indices were 100.0%, 100.0%, 100.0% and
91.7% in the same respective groups

Details on results (P0)

Mean gestation lengths in the 150, 450 and 1000 mg/kg/day groups were similar to those
in the control group. No statistically significant differences were noted. No signs of
dystocia were noted in these groups.

Effect levels (P0)

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

mortality

body weight and weight gain

food consumption and compound intake

organ weights and organ / body weight ratios

gross pathology

reproductive function (oestrous cycle)

reproductive function (sperm measures)

reproductive performance

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Description (incidence and severity):

There were no test article-related effects on the mean numbers of pups born, live litter
size on PND 0, the percentage of males at birth or postnatal survival at any dosage level;
no statistically significant differences from the control group were observed. While the
mean numbers of pups born and live litter size on PND 0 in the test article-treated groups
were slightly lower than concurrent control group values, the values were within the
range of the WIL historical control data.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

The numbers of F1 pups found dead and/or missing, as well as the general physical
condition of all F1 pups in this study were unaffected by parental test article
administration. Pups (litters) that were found dead numbered 5(4), 4(3), 7(3) and 2(2) in
the control, 150, 450 and 1000 mg/kg/day groups, respectively. One and 3 pups in the
control and 150 mg/kg/day groups, respectively, were missing and presumed to have
been cannibalized.
The numbers of pups (litters) found dead during PND 0-4 numbered 5(4), 4(3), 7(3) and
2(2) in the control, 150, 450 and 1000 mg/kg/day groups, respectively. Three and 5 pups
in the control and 450 mg/kg/day groups, respectively, had lungs that did not float and
were considered stillborn. Aside from the presence or absence of milk in the stomach, no
other internal findings were noted.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Mean male and female pup body weights and body weight changes in the 150, 450 and
1000 mg/kg/day groups were unaffected by test article administration during PND 1-4.
No statistically significant differences from the control group were noted.

Food consumption and compound intake (if feeding study):

not examined

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

Sexual maturation:

not examined

Anogenital distance (AGD):

not examined

Nipple retention in male pups:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

Other effects:

not examined

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 1 000 mg/kg bw/day

Based on:

test mat.

Sex:

not specified

Basis for effect level:

viability

clinical signs

mortality

body weight and weight gain

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Lowest effective dose / conc.:

1 000 mg/kg bw/day

Treatment related:

no

Applicant's summary and conclusion

Conclusions:

Under the conditions of this study, EC 701-204-9 did not exhibit adverse reproductive or developmental effects and the NOAEL >1000 mg/kg/day (the highest dose tested)

Executive summary:

The registered substance was administered orally (gavage) in corn oil to 3 groups of Crl:CD(SD) rats, each group consisting of 12 males and 12 females. Dosage levels were 150, 450 and 1000 mg/kg/day administered at a dosage volume of 5 mL/kg. A concurrent control group of 12 rats/sex received the vehicle (corn oil) on a comparable regimen. Males received 14 daily doses prior to mating. Males were dosed throughout the mating period through 1 day prior to euthanasia for a total of 43 doses. Females received 14 daily doses prior to pairing and were dosed through lactation day 3 for a total of 39-43 doses; the female with no evidence of mating was dosed through the day prior to euthanasia (post-cohabitation day 25) for a total of 52 doses. Under the conditions of this screening study, there were no effects on survival, body weights, food consumption, organ weights, macroscopic or microscopic evaluations and functional reproductive outcome in the F0 males and females at any dosage level, a dosage level of 1000 mg/kg/day, the highest level tested, was considered to be the no observed-adverse-effect level (NOAEL) for systemic and reproductive toxicity of 68784 -17-8 when administered orally by gavage to Crl:CD(SD) rats. The NOAEL for neonatal toxicity was 1000 mg/kg/day based on the lack of effects on postnatal survival, physical condition and body weights at any dosage level.