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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:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Supplier: Aldrich Chemical Company
- Batch: 61496MK

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (Crl:CD (SD) IGS BR)
- Age at study initiation: approximately 10 to 12 weeks of age at receipt on GD 1 to 3
- Weight at study initiation: The weight range was 245 to 299 grams at start of dosing
- Fasting period before study: No
- Diet: Certified Global 16% Protein Rodent Diet No. 2016C (pellet, Harlan), ad libitum
- Water: Water (New Jersey-American Water Company), ad libitum
- Acclimation period: The study animals arrived at the test facility on GD1-3 and were acclimated until GD6.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Daily Average Range, 20 to 23°C
- Humidity (%): Daily Average Range, 47 to 53%
- Air changes (per hr): no information

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: A dosing volume of 5 mL/kg was applied for all animals, which was adjusted based on the latest body weight. No neutralization of the dose formalations was performed. Fresh dose formulations were prepared once daily. The pure test article has a pH of 1.1 and pKa of 0.3, and reacts strongly with water. Therefore, a PAPR with an OV/SD/HV/CL/HE organic vapour filter was used during preparation and dose administration. Due to evaporation properties of the formulations, the containers were covered with parafilm or similar covering during preparation, as well as, during dosing to ensure accurate and consistent concentration results.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analyses to determine homogeneity, stability, and concentration of the test and/or control materials were carried out at the testing facility.
Details on mating procedure:
The female rats were inseminated at the vendor facility. Gestation Day 0 (GD0) was the day of detection of a copulatory plug in situ and/or sperm in a vaginal smear. The weight range was 245 to 299 grams at start of dosing
Duration of treatment / exposure:
Gestation day 6 up to and including gestation day 19
Frequency of treatment:
Daily
Duration of test:
14 treatment days.
Doses / concentrationsopen allclose all
Dose / conc.:
37.5 mg/kg bw/day (nominal)
Dose / conc.:
75 mg/kg bw/day (nominal)
Dose / conc.:
150 mg/kg bw/day (nominal)
No. of animals per sex per dose:
22 mated female rats
Control animals:
yes, concurrent vehicle

Examinations

Maternal examinations:
VIABILITY:
Viability (morbidity and mortality) checks were made twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: detailed physical examinations were performed immediately prior to dosing [within approximately 30 minutes] and approximately 2 hours after dosing.

BODY WEIGHT: Yes
- Time schedule for examinations: GD4, and once daily from GD6-20

FOOD CONSUMPTION: Yes
- Time schedule for examinations: GD4-20 (2- or 3-days consumptions)

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: A macroscopic post-mortem evaluation was performed on GD 20, during which corpora lutea, implantation data and uterine weights were recorded. In addition, liver and kidney weights were determined.
Ovaries and uterine content:
The ovaries and uteri were examined to determine:
- Number of corpora lutea;
- Weight of intact gravid uterus;
- Location and number of implantation sites, resorptions, fetal death, dead fetuses;
- Gross evaluation and weight of placentas.
Fetal examinations:
The fetuses were removed, weighed (live and recently dead fetuses), sexed and examined externally for defects. Approximately half of the fetuses were examined for soft tissue abnormalities. The other half was processed for staining with Alizarin Red S and examined for skeletal abnormalities and ossification state.
Statistics:
Statistical analyses were performed using the individual animal (or litter) as the basic experimental unit. For litter/fetal findings, the litter was the treated experimental unit and the basis for statistical analysis and biological significance was assessed with relevance to the severity of the anomaly and the incidence of the finding within the background control population.The following statistical methods have been used for the analysis of the results: Williams' test, Dunnett's test, Shirley's test, Steel's test, Kruskal-Wallis test, Wilcoxon rank sum tests
Indices:
Sex ratio, Pre implantation loss, Post implantation loss, Gravid uterine adjusted body weights, Mean litter placental and fetal weights

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
no effects observed
Description (incidence and severity):
.
Mortality:
no mortality observed
Description (incidence):
All females survived until termination.
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
Description (incidence and severity):
Test article-related organ weight increases were observed in liver and kidney (see Table below in 'any other information on results')
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined

Maternal developmental toxicity

Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
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
Description (incidence and severity):
One female each in the control and high-dose groups was not pregnant, although this was not attributed to the administration of the test article.

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
150 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: No treatment related effects on maternal toxicity at the highest dose level

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

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
150 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No treatment related effect on embryo-fetal development in rats.

Fetal abnormalities

Abnormalities:
no effects observed

Overall developmental toxicity

Developmental effects observed:
no

Any other information on results incl. tables

Analysis of initial mixes confirmed that the preparation procedure used for this study produced homogeneous mixtures and that the test material was stable in the vehicle, under refrigerated conditions, for at least seven days. The formulations were prepared daily for dosing, and no storage was required. Analyses conducted during the treatment period confirmed that dose solutions of appropriate concentration were administered.

Body weight in pregnant rats (body weight on GD20 was adjusted by subtracting weight of uterus with fetuses):

 

Body weight (g)

Kidney (g)

Liver (g)

Dose (mg/kg)

GD4

GD6

GD12

GD16

GD20

GD20-adjusted

0

264

284

310

336

383

309

 1.97

15.6 

37.5

265

284

310

339

376

306

2.05

15.7

75

261

278

303

333

384

299

2.05

15.7

150

266

282

307

337

386

308

2.08*

17.1*

Number of live fetuses, mean number of live fetuses per litter and mean body weight of fetuse:

Dose (mg/kg)

Fetuses/Litters

Fetuses per litter

Body weight (g)

0

244/21

12

4.0

37.5

279/22

13

4.1

75

294/22

13

4.0

150

251/21

12

4.0

Applicant's summary and conclusion

Conclusions:
The No-Observed-Adverse-Effect-Level (NOAEL) of Trifluoroacetic acid for maternal and embryo-fetal development toxicity in rats was considered to be 150 mg/kg/day.
Executive summary:

A GLP compliant prenatal developmental toxicity study with Trifluoroacetic acid was conducted in Sprague Dawley rats according to OECD Guideline 414. Mated females were treated with Trifluoroacetic acid by oral gavage at dose levels of 37.5, 75, and 150 mg/kg/day during gestation (from Gestation Day 6 to 19). A control group of 22 females was included and the animals were given the vehicle (distilled water).

The study animals were observed daily for mortality and clinical signs. On GD 20, all the surviving study animals were necropsied and examined macroscopically. The ovaries and uteri were examined for determination of litter data. The fetuses were removed, weighed (live and recently dead fetuses), sexed and examined externally for defects. All the live fetuses were weighed and examined for external abnormalities; their crown-rump length was measured and their sex was determined. Approximately half of the fetuses were examined for soft tissue abnormalities. The other half was examined for skeletal abnormalities and ossification state.

All females survived until termination. Dosing was well-tolerated by all females, and doses up to 150 mg/kg/day had no adverse effect on body weight, body weight gain, food consumption, pregnancy, c-section parameters, fetal, placental, and uterine weights, organ weights, or fetal abnormalities, variations or ossification parameters.

Based on these results, the No-Observed-Adverse-Effect-Level (NOAEL) of Trifluoroacetic acid for maternal and embryo-fetal development toxicity in rats was considered to be 150 mg/kg/day. Due to the non-adverse, test article-related organ weight increases, the maternal and embryo-fetal no-observed-effect-levels (NOEL) were established at 75 mg/kg/day (maternal) and 150 mg/kg/day (embryo-fetal).