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Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD guideline conducted under GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Qualifier:
according to
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approx. 10 - 11 weeks old
- Housing: macrolon cages
- Diet: ad libitum except during exposure
- Water: ad libitumexcept during exposure
- Acclimation period: at least 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2°C
- Humidity (%): 40-70%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light):12/12

Administration / exposure

Route of administration:
inhalation: gas
Type of inhalation exposure (if applicable):
nose only
Vehicle:
unchanged (no vehicle)
Details on exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: nose only exposure units, cylindrical PVC column with volume of ~ 70 liters, surrounded by a transparent hood
- Source and rate of air:humidified compressed air
- System of generating particulates/aerosols: not applicable
- Temperature, humidity in air chamber: 21.9 - 23.1 degrees Celcius; 34-45% humidity;
- Method of particle size determination: not applicable
- Treatment of exhaust air: not described

TEST ATMOSPHERE
- Brief description of analytical method used: total carbon analysis
- Samples taken from breathing zone: yes
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
total carbon analysis with flame ionization detector
Details on mating procedure:
- Impregnation procedure: [cohoused]
- If cohoused:
- M/F ratio per cage: 1/2
- Length of cohabitation:
- After 7 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: not necessary
- Verification of same strain and source of both sexes: [yes / no (explain)]
- Proof of pregnancy: [ sperm in vaginal smear] referred to as [day 0 ) of pregnancy
- Any other deviations from standard protocol:
Duration of treatment / exposure:
gestation day 6 - 19
Frequency of treatment:
6 hours/day
Doses / concentrations
Remarks:
Doses / Concentrations:
5000, 15000, 50000 ppm
Basis:
nominal conc.
No. of animals per sex per dose:
25
Control animals:
yes, sham-exposed
Details on study design:
- Dose selection rationale: Doses used were similar to those used for other inhalation repeated dose studies conducted on this test material.As noted in paragraph 13 of OECD guideline 414, the dose levels should be selected taking into account any existing toxicity. The highest dose tested in the this study (15,000 ppm) was based on the cardiac effects observed in the 2 week, 4 week, and 13 week toxicity studies conducted in rats. As noted in the table attached in the background info section. In the 2 wk, 4 week, and 90 day inhalation toxicity studies, cardiac effects including mononuclear cell infiltrates were observed at of 15000 – 20000 ppm and higher. In these studies, there were also no clinical signs of toxicity. The rat prenatal toxicity study did not include evaluation of the cardiac tissue as this is not a standard endpoint for these studies and data were available on this endpoint from the repeated dose studies. Honeywell considers the existing rat prenatal toxicity study to be compliant with the REACH regulation.

Table 1- Summary of effects seen in the heart of rats exposed to HFO-1234ze1
Study / effects Males Females
2-Week exposure levels in 1000 ppm 0 5 20 50 0 5 20 50
Myocardial vacuolation
Slight 0 0 1 3 0 0 0 1
Moderate 0 0 0 2 0 0 0 4
Total 0 0 1 5** 0 0 0 5**
Pyknosis
Very slight 0 0 1 5 0 0 0 4
Slight 0 0 0 0 0 0 0 1
Total 0 0 1 5** 0 0 0 5**
Mononuclear cell infiltrate
Very slight 1 0 0 4 0 3 0 2
Slight 0 0 1 0 0 0 0 2
Moderate/multifocal 0 0 4 0 0 0 5 0
Total 1 0 5* 4 0 3 5** 4*

4-Week exposure levels in 1000 ppm 0 1 5 10 15 0 1 5 10 15
Myocardial vacuolation 0 0 0 0 2 0 0 0 0 0
Myocardial vacuolation,
not associated with inflammation 0 0 0 0 0 0 0 1 0 0
Mononuclear cell infiltrate
Very slight 0 1 1 1 3 0 0 2 1 2
Slight 0 0 0 0 1 0 1 0 0 0
Moderate/multifocal 0 0 0 0 1 0 0 0 0 0
Total 0 1 1 1 5** 0 1 2 1 2

13-Week exposure levels in 1000 ppm 0 1.5 5 15 0 1.5 5 15
Very slight to slight focal
mononuclear cell infiltrate 4 6 6
1 3 5 6 4
Multifocal mononuclear cell infiltrate
Very slight 0 0 0 1 0 0 0 1
Slight 0 0 0 7 0 0 0 4
Moderate 0 0 0 1 0 0 0 0
Totals 0 0 0 9*** 0 0 0 5*

*p < 0.05; **p < 0.01; ***p < 0.001
1 N= 5 animals/sex/group in the 2-week and 4-week exposure studies; N=10 animals/sex/group in the 13-week exposure study

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS for clinical signs and mortality: Yes, daily

BODY WEIGHT: Yes (GD 0,3,6,9,12,15,19,and 21)

FOOD CONSUMPTION: Yes (GD 0-3, 3-6, 6-9, 9-12, 12-15, 15-19, and 19-21)
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:
- Soft tissue examinations: Yes: half per litter
Skeletal examinations: Yes: half per litter
Statistics:
Fisher's exact probability test, one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison tests or Kruskal-Wallis
nonparametric analysis of variance followed by the Mann-Whitney U-test as appropriate. Statistical evaluations on variables associated with the fetuses were considered on a litter basis in accordance to standard procedures.
Indices:
Female fecundity index, Gestation index

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
No mortality was seen. No remarkable clinical signs were observed compared to controls. No effect observed on mean maternal body weight during pregnancy (however, body weight loss was noted in all groups including controls were noted during GD 6-9 and 19-21). The mean body weight change during the total exposure was comparable in all groups. No effects were observed in food comsumption. No differences were noted in number of females mated, number pregnant at C-section, female fecundity index, corpora lutea, implantation sites, pre or post implanation loss, live fetuses, dead fetuses, resorptions (total, early or late). Weights of gavid uterus, carcass, empty uterus, and ovaries were not different from controls. Macroscopic findings at necropsy did not reveal any remarkable or treatment related findings.

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOEC
Effect level:
15 000 ppm (analytical)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEC
Effect level:
15 000 ppm (analytical)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No statistically significant differences in the incidence of external observations were observed among the groups. Enlarged placenta was seen in all live fetusus in one high dose dam but no abnormalities were observed when these placentas were evaluated micropscopically. Fetal and placental weights were not different. No statistically significant differences were noted in the incidence of visceral malformations, anomolies, or variations. Skeletal examinations did not show a statistically significant difference in incidence of skeletal malformations, anomalies, or variation were observed among the groups. Although there were a few cases of statistically significant changes in ossification, they are not always dose related and often show a decrease in delayed or incomplete ossification with dose. There was no difference in total fetaly skeletal retardations based on fetal or litter incidence. The effects observed are not considered to be adverse.

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
In conclusion, under the conditions of this study, HFO-1234ze given by inhalation to pregnant female rats from gestation day (GD) 6-20 for 6 hours per day up to 15,000 ppm did not induce maternal or prenatal developmental toxicity. Therefore this level represents the NOAEL for developmental effects in this study.