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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: This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labelling and/or risk assessment.
Cross-reference
Reason / purpose for cross-reference:
reference to same 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
Guideline:
other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
Study included FOB and motor activity endpoints.
Qualifier:
according to guideline
Guideline:
other: EPA OPPTS 870.3650 (2000)
Deviations:
yes
Remarks:
Study included FOB and motor activity endpoints.
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Reference substance name:
Lecithins, acetylated
EC Number:
293-316-5
EC Name:
Lecithins, acetylated
Cas Number:
91053-50-8
Molecular formula:
Not applicable- complex UVCB substance
IUPAC Name:
91053-50-8
Details on test material:
Purity: Unknown variable composition biological substance (UVCBS)
Composition of test material, percentage of components: Unknown variable composition biological substance (UVCBS)

Test animals

Species:
rat
Strain:
other: Crl:CD(SD)
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: approximately 65 days
- Weight at study initiation: 300-357 grams (males) and 215-260 grams (females)
- Housing: Solid-bottom cages contained bedding material, and a nestlet and nylabone for enrichment. Sexes were placed on separate racks (except during mating).
- Diet (e.g. ad libitum): ad libitum except when fasted
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Approximately 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26ºC
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): approximate 12-hour light/dark cycle
- Air Changes: no data

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing formulations were prepared by mixing the test substance into the vehicle control substance. Formulations were prepared daily. Formulations were stored at room temperature.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test Formulation Sampling: Near the beginning of the study, dose formulations were sampled and analyzed for verification of concentration, homogeneity, and stability. Near the middle and end of the study, dose formulations were sampled for an additional concentration verification check.

Method of Analysis: Analysis was performed by 31P NMR Spectroscopy. Samples were shipped on the day they were collected at ambient temperature for NMR analysis. At the time of analysis, the samples were diluted with an appropriate solvent and analyzed by 31P Nuclear Magnetic Resonance Spectroscopy.

Analyses results show that the test substance was homogeneously mixed at the targeted concentrations, and stable in the vehicle under the storage conditions for all dose levels for the study. Test substance was not detected in the control samples.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: until evidence of mating was observed or until a period of 2 weeks had elapsed
- Proof of pregnancy: Copulation plug in situ or sperm in vaginal lavage sample referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged individually. Females that did not mate at the end of the 2-week period were transferred to individual cage housing.
Duration of treatment / exposure:
All animals were dosed once daily by gavage for approximately 14 days prior to cohabitation and during the cohabitation period (up to 2 weeks). Male and female rats showing no evidence of copulation continued to be dosed after the end of the cohabitation period until sacrifice. Females showing evidence of copulation were dosed throughout gestation. Pregnant females in the process of delivery or showing signs of delivery were not dosed. Females were dosed after delivering litters, until day 3 postpartum. Females that did not deliver a litter continued to be dosed until the day before sacrifice.
Frequency of treatment:
Daily
Duration of test:
Approximately 7 weeks
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 100, 300, 1000 mg/kg
Basis:
other: actual administered to parents. Offspring only exposed through nursing
No. of animals per sex per dose:
12 animals/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In a previous acute oral toxicity study (up and down procedure) in which the test substance was dosed neat, the LD50 value was determined to be > 5000 mg/kg in female rats. Based on this data, doses of 100, 300, and 1000 mg/kg/day were selected for the current study. These dose levels include the limit dose level of 1000 mg/kg/day as specified by the relevant testing guidelines. The low and intermediate dose levels were selected to enable detection of potential dose responses across a wide range of dose levels.

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once daily, at least 2 hours post-dosing

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Prior to dosing on test day 0 (baseline) and weekly thereafter at approximately the same time of day (± 2 hours); mortality and moribundity check twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: Prior to dosing on test day 0 and weekly during premating and postmating; gestation days (GD) 0, 7, 14, and 20 and lactation days (LD) 0 and 4 and at neurobehavioral evaluations and scheduled sacrifice

FOOD CONSUMPTION:
- Food consumption for each animal determined (g/food/animal/day): Yes; Weekly during premating GD 0, 7, 14, and 20 and LD 0 and 4. Food consumption was not measured during the postmating period for males, and females that did not mate.

FOOD EFFICIENCY:
- Calculated as g weight gained/g food consumed: Yes

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Test day 14
- Anaesthetic used for blood collection: Yes; Carbon dioxide or Isoflurane
- Animals fasted: Yes
- How many animals: 5/sex/group
- Parameters in table No. 1 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Test day 14
- Animals fasted: Yes
- How many animals: 5/sex/group
- Parameters in table No. 2 were examined.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes (refer to Section 7.9.1, DI.K1.Neuro.D-18511-1422 for additional details).

OTHER: Coagulation
- Time schedule for collection: at sacrifice
- How many animals: 5/sex/group
- Anaesthetic used: Carbon dioxide

GROSS PATHOLOGY: Yes (see table 3)
HISTOPATHOLOGY: Yes (see table 3)

Gross examinations were performed on all adult rats. Final body and organ weights were recorded for all rats euthanized at the scheduled sacrifice. Microscopic examination of all reproductive organs was conducted for all rats in the control (0 mg/kg/day) and high-treatment (1000 mg/kg/day) groups and for all reproductive failures (male and female pairs). One mated pair (300 mg/kg/day) required a caesarean section and was also evaluated microscopically. Microscopic examination of all remaining tissues collected was limited to the first 5 surviving rats in the males and female high-treatment and control groups. All collected tissues were also examined microscopically for the 2 decedents. Most gross lesions recorded at necropsy were evaluated microscopically. Gross lesions which were unlikely to have a microscopic correlate (e.g., wet, stain) were not examined.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Pre-Implantation loss: Yes
-Post-implantation loss: Yes
Fetal examinations:
Each litter was examined as soon as possible after delivery was completed. The day when delivery was complete was designated lactation day (LD) 0 for dams and postnatal day (PND) 0 for pups. On LD 0 and 4 (day of litter sacrifice) live and dead pups in each litter were counted by sex and live pups were individually weighed. Each live pup was individually handled and examined for abnormal behaviour and an external examination was conducted. Dead pups were examined externally to the extent possible and underwent a gross post-mortem examination.
Statistics:
Statistical analysis was conducted such that data from each group were compared to the vehicle control group using an appropriate pairwise analysis (see Table 4). Male and female data were evaluated separately. For litter parameters, the proportion of affected pups per litter or the litter mean were used as the experimental unit for statistical evaluation. The level of significance selected was p <0.05. For each parameter analyzed with a trend test, the test was applied to the data sequentially. If a significant dose-response was detected, data from the top dose group was excluded and the test repeated until no significant trend was detected.
Indices:
Refer to Table 5 for indices calculated.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

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

Details on maternal toxic effects:
Body weight and body weight gains: At 1000 mg/kg/day, mean body weight gain was significantly lower than controls for the last week of gestation (GD14-20). This reduction was not considered adverse since it did not impact mean body weight at the end of gestation which was within 4% of the control group mean.

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Basis for effect level:
other: maternal toxicity

Results (fetuses)

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

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
NOAEL = 1000 mg/kg/bw/day
Executive summary:

The objective of this study was to evaluate the potential subchronic and reproductive/developmental toxicity of the test substance when administered by oral gavage to male and female rats during premating, cohabitation, gestation, until postnatal day (PND) 3. Four groups of parental (P0) male and female Crl:CD(SD) rats (12/sex/group) were administered formulations of the test substance via once daily oral gavage for 14 consecutive days prior to mating, throughout mating, and then continuing through one day prior to the day of euthanasia. The dose levels administered were 0, 100, 300, or 1000 mg/kg/day. Samples of the dose formulations were analyzed and confirmed that the formulations were at targeted concentrations, homogeneous, and stable under the conditions of use.  During the in-life phase of the study, all animals were observed twice daily for appearance and behaviour. Clinical observations, body weights, and food consumption were recorded at appropriate intervals for Pmales throughout the study and for Pfemales prior to mating and during gestation and lactation. Neurobehavioral examinations consisting of an abbreviated functional observational battery (FOB) and motor activity (MA) were performed for all P0 animals once prior to dosing to provide baseline data and once more near the end of the premating period. In addition, clinical pathology data consisting of haematology, coagulation, and clinical chemistry data were collected prior to the end of the premating period. All Pfemales were allowed to deliver and rear their pups until weaning on PND 4. Each Pparental animal received a complete detailed gross necropsy and selected organs were weighed and/or retained for histopathologic examination.

 

There was no evidence of test substance-related toxicity at any dose level tested. The in-life data, including body weight and food consumption parameters, clinical observations, neurobehavioural endpoints (FOB and MA), clinical pathology, as well as reproductive performance data including mating and fertility indices, gestation length, offspring viability, litter sizes, litter sex ratio, and pup weights, were comparable across all groups throughout the study. In addition, there were no test substance-related effects on either gross or microscopic histopathology or on organ weights. 

 

Under the conditions of this study, the no-observed-adverse-effect level (NOAEL) for developmental/reproductive toxicity was 1000 mg/kg/day. This conclusion is based on the lack of any evidence of any adverse and/or test substance-related effects at any dose level tested.