Registration Dossier

Administrative data

Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2008-01-07 till 2008-04-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study under GLP without deviations

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Milled SLI (76)
- EC name of test material: Fatty acids, C12-18 and C18-unsatd., 2-sulfoethyl esters, sodium salts
- Physical state: Off-white fine powder
- Storage condition of test material: At room temperature (20 ± 5 °C) in the original container away from direct sunlight, with dessicant

Test animals

Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd., Füllinsdorf / Switzerland
- Age at study initiation: 11 weeks
- Weight at study initiation: 187 to 237 g
- Fasting period before study: no data
- Housing: Individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Kliba Nafag 3433 rat/mouse maintenance diet, ad libidum
- Water (e.g. ad libitum): Community tap-water from Füllinsdorf, ad libidum
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
For groups 2 and 3: every second week
For group 4: daily.

DIET PREPARATION:
not applicable (administration by gavage)


VEHICLE
- Concentration in vehicle: no data
- dose volume applied: 10 mL/kg
- Amount of vehicle (if gavage): 10 mL/kg
- Purity: Milli-Q-water
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
For groups 1, 2 and 3:
The formulations mixed for the first two weeks of the study were sampled to confirm achieved concentration and homogeneity. One ml samples were taken in triplicate from the top, middle and bottom of the treated formulation during stirring. Five 1 mL samples were taken from the vehicle control group. After dosing on day 1 of each preparation, the residual formulation were stored frozen. From the formulations mixed on each subsequent occasion, five 1 mL samples were taken from the middle of each formulation during stirring and from the vehicle control. These samples were used to confirm achieved concentration only.
For group 4:
The formulation mixed for the first day of treatment was sampled to confirm achieved concentration and homogeneity. One ml samples were taken in triplicate from the top, middle and bottom of the formulation during stirring. Thereafter every two weeks samples were taken to confirm achieved concentration. Five 1 mL samples were taken from the middle of the formulation during stirring on each occasion.

All formulation samples (for all groups) despatched to the Principal Investigator were transferred to an amber glass vial and frozen (-20 ± 5 °C) pending analysis by Liquid Chromatography/Mass Spectrometry (LC/MS).
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: until evidence of copulation was observed
- Further matings after two unsuccessful attempts: no data
- Verification of same strain and source of both sexes: no (these male rats were in the possession of RCC)
- Proof of pregnancy: vaginal plug or sperm in vaginal smear, referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no
Duration of treatment / exposure:
from day 6 post-coitum to day 20 post-coitum
Frequency of treatment:
Daily
Duration of test:
20 days (day 21: females sacrificed and fetuses removed by Caesarean section)
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
100 mg/kg bw/day
Basis:

Remarks:
Doses / Concentrations:
300 mg/kg bw/day
Basis:

Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:

No. of animals per sex per dose:
22 females per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected based on a previous Reproduction/Developmental Toxicity Screening Test in the Han Wistar Rat, RCC Study Number B57982, using dose levels of 100, 300, and 1000 mg/kg/day, which showed no adverse effects on reproduction.

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily


DETAILED CLINICAL OBSERVATIONS: No data


BODY WEIGHT: Yes
- Time schedule for examinations: daily from day 0 until day 21 post coitum


FOOD CONSUMPTION: Yes
- Time schedule: recorded at 3-day intervals: days 0-3, 3-6, 6-9, 9-12, 12-15, 15-18 and 18-21 post coitum.


POST-MORTEM EXAMINATIONS: Yes (gross macroscopic examination)
- Sacrifice on gestation day 21 post coitum
- Organs examined: all internal organs with emphasis to on the uterus, uterine contents, position of fetuses in the uterus and the number of corpora lutea

Ovaries and uterine content:
The uterine content was examined after termination: Yes
The ovaries content examined after termination: No
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:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter
Statistics:
- Means and standard deviations of various data
- The Dunnett-test (many to one t-test) based on a pooled variance estimate, applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-one rank test), applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test, applied if the variables could be dichotomized without loss of information.
Indices:
No data
Historical control data:
Yes

Results and discussion

Results: maternal animals

Maternal developmental toxicity

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

Details on maternal toxic effects:
General Tolerability - All females survived until the scheduled necropsy. No signs of discomfort or clinical symptoms from the treatment with the test item were observed.
Food Consumption and Body Weights - Mean food consumption, mean body weight and corrected body weight gain (corrected for the gravid uterus weight) were not affected by treatment with the test item in any dose group.
Reproduction Data - Post-implantation losses and the mean number of fetuses per dam were not affected by treatment with the test item at any dose level.
Macroscopical Findings - No macroscopical findings were noted during necropsy of the females.

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day
Basis for effect level:
other: developmental toxicity

Results (fetuses)

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

Details on embryotoxic / teratogenic effects:
External Examination - During the external examination of the fetuses, no test item-related abnormal findings were noted.
Sex Ratios - No test item-related effects on fetal sex ratios were noted in any dose group.
Body Weights - No test item-related effects on fetal body weights were noted.
Visceral Examination - No test item-related abnormalities were noted during the visceral examination of fetuses.
Skeletal and Cartilage Examination - No abnormalities, which were considered to be test item-related, were noted during examination of fetal skeletons and cartilages.

Effect levels (fetuses)

Remarks on result:
not determinable due to absence of adverse toxic effects

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Any other information on results incl. tables

Maternal Toxicity:

General Tolerability

All females survived until the scheduled necropsy. No signs of discomfort or clinical symptoms from the treatment with the test item were observed.

Food Consumption and Body Weights

Mean food consumption, mean body weight and corrected body weight gain (corrected for the gravid uterus weight) were not affected by treatment with the test item in any dose group.

Reproduction Data

Post-implantation losses and the mean number of fetuses per dam were not affected by treatment with the test item at any dose level.

Macroscopical Findings

No macroscopical findings were noted during necropsy of the females.

Embryotoxic/teratogenic effects:

External Examination

During the external examination of the fetuses, no test item-related abnormal findings were noted.

Sex Ratios

No test item-related effects on fetal sex ratios were noted in any dose group.

Body Weights

No test item-related effects on fetal body weights were noted.

Visceral Examination

No test item-related abnormalities were noted during the visceral examination of fetuses.

Skeletal and Cartilage Examination

No abnormalities, which were considered to be test item-related, were noted during examination of fetal skeletons and cartilages.

Applicant's summary and conclusion

Conclusions:
Based on the above mentioned results, the NOEL (No Observed Effect Level) for maternal and fetal organisms was considered to be 1000 mg/kg body weight/day.
Under the conditions described for this study, MILLED SLI (76) did not reveal teratogenic potential up to and including 1000 mg/kg body weight/day.
Executive summary:

Females were treated with MILLED SLI (76) to detect effects on the pregnant rat and development of the embryo and fetus.

Four groups of females were treated by gavage once daily (from day 6 post coitum to day 20 post coitum) at dose levels of 0 mg/kg body weight/day (control group), 100 mg/kg body weight/day, 300 mg/kg body weight/day and 1000 mg/kg body weight/day. A standard dose volume of 10 mL/kg body weight with a daily adjustment to the actual body weight was used. Control animals were dosed with the vehicle alone (Milli-Q-Water).

All females survived until the scheduled necropsy. No signs of discomfort or clinical symptoms from the treatment with the test item were observed. Mean food consumption, mean body weight and corrected body weight gain (corrected for the gravid uterus weight) were not affected by treatment with the test item in any dose group. Post-implantation losses and the mean number of fetuses per dam were not affected by treatment with the test item at any dose level. No macroscopical findings were noted during necropsy of the females.

No test item-related abnormal findings were noted during the external examination of the fetuses. No test item-related effects on fetal sex ratios were noted in any dose group. No test item-related effects on fetal body weights were noted. No test item-related abnormalities were noted during the visceral examination of fetuses. No abnormalities, which were considered to be test item-related, were noted during examination of fetal skeletons and cartilages.

Based on the above mentioned results, the NOEL (No Observed Effect Level) for maternal and fetal organisms was considered to be 1000 mg/kg body weight/day.