Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Developmental toxicity / teratogenicity

Currently viewing:

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 Apr - 28 Nov 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 Apr - 28 Nov 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
adopted 22. March 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EPA OPPTS 870.3650
Version / remarks:
July 2000
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
- Physical state: liquid
- Analytical purity: 87%
- Active content: 30.89%
- Lot/batch No.: 184
- Expiration date of the lot/batch: 30 June 2008
- Storage condition of test material: room temperature (20 ± 5°C)
Species:
rat
Strain:
Wistar
Remarks:
HanRcc: WIST(SPF)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories Ltd., Switzerland
- Age at study initiation: 11 weeks
- Weight at study initiation: mean: males 294 - 344 g; females 176 - 221 g
- Housing: individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding
- Diet: Pelleted standard Kliba Nafag 3433 rat/mouse maintenance diet, ad libitum
- Water: community tap-water, ad libitum
- Acclimation period: 7 days

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

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
highly purified
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The dosage formulations were prepared weekly prior to administration using the test item as supplied by the Sponsor and using a factor of 3.72 taking in account the purity of 87% and the content of the active ingredient of 30.89% (CAS 3655-00-3). Sodium coco β-iminodipropionate (CAS 3655-00-3) was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration. Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Several application formulations were prepared and representative analytical samples were collected and dispatched to the analytical laboratories internally. The test item concentrations were determined by HPLC coupled to an ELSD detector and quantified with the area under the peak.
Duration of treatment / exposure:
Males: minimum of 4 weeks (2 weeks prior to mating and 2 weeks of mating)
Females: approximately 7 weeks (2 weeks prior to mating up to day 4 post partum)
Frequency of treatment:
daily, 7 days/week
Dose / conc.:
43 mg/kg bw/day (actual dose received)
Remarks:
Group 2
Dose / conc.:
160 mg/kg bw/day (actual dose received)
Remarks:
Group 3
Dose / conc.:
600 mg/kg bw/day (actual dose received)
Remarks:
Group 4
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dosage levels were selected based on a previous dose range finding toxicity study in Han Wistar Rats.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS
- Time schedule: daily
- Cage side observations checked: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern), changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported

BODY WEIGHT
- Time schedule for examinations: daily from treatment start to day of necropsy

FOOD CONSUMPTION AND COMPOUND INTAKE
- was recorded for male animals weekly during pre-pairing, for females weekly in during the pre-pairing period, gestation days 0-7, 7-14 and 14-21 post coitum, and days 1-4 post partum; food consumption was not recorded during the pairing period

HAEMATOLOGY
- Time schedule for collection of blood: on day before or day of necropsy for males, for females on day 5 post partum
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- Number of animals: 5 males, 5 females
- Parameters examined: Complete Blood Cell Count, Erythrocyte count, Hemoglobin, Hematocrit, Mean corpuscular volume, Red cell volume distribution width, Mean corpuscular hemoglobin, Mean corpuscular hemoglobin concentration, Hemoglobin concentration distribution width, Leukocyte count (total), Differential leukocyte count, Platelet count, Coagulation, Prothrombin time (= Thromboplastin time), Activated partial Thromoplastin time

CLINICAL CHEMISTRY
- Time schedule for collection of blood: on day before or day of necropsy for males, for females on day 5 post partum
- Animals fasted: Yes
- Number of animals: 5 males, 5 females
- Parameters examined: Glucose, Urea, Creatinine, Bilirubin (total), Cholesterol (total), Triglycerides, Aspartate aminotransferase, Alanine aminotransferase, Alkaline phosphatase, Gamma-glutamyl-transferase, Bile acids, Creatine kinase, Sodium, Potassium, Chloride, Calcium, Phosphorus, Protein (total), Albumin, Globulin, Albumin/Globulin ratio

NEUROBEHAVIOURAL EXAMINATION
- Time schedule for examinations: males shortly before the scheduled sacrifice and females on day 3 or 4 post partum
- Dose groups that were examined: each group
- Battery of functions tested: sensory activity / grip strength / motor activity / other: cage-side and hand-held observations
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

ORGAN WEIGHTS
- testes and epididymides of all parental males were weighed as pairs, from 5 males and females selected randomly from each group, the following organs
were trimmed from any adherent tissue, as appropriate, and their wet weight taken: Adrenal glands (weighed as pairs), Brain, Heart, Kidneys (weighed as pairs), Liver, Thymus, Spleen

HISTOPATHOLOGY
- Parental males tissues (preserved in neutral phosphate buffered 4% formaldehyde solution): Prostate, Seminal vesicles with coagulating gland, Testes (in Bouin’s fixative), Epididymides (in Bouin’s fixative)
- Parental females tissues (preserved in neutral phosphate buffered 4% formaldehyde solution): Ovaries
- Tissues (preserved in neutral phosphate buffered 4% formaldehyde solution) from the 5 male and female animals per group selected for organ weights: Gross lesions, Brain (cerebrum, cerebellum, pons), Spinal chord, Small and large intestines (incl. Peyer’s patches), Stomach, Liver, Kidneys, Adrenals, Spleen, Heart, Thymus, Thyroid and Parathyroid, Trachea and lungs (preserved by inflation with fixative and then immersion), Uterus (with vagina), Urinary bladder, Lymph nodes (mandibular and mesenteric), Peripheral nerve (sciatic), Bone marrow
Statistics:
The following statistical methods may be used to food consumption, body weight, macroscopic findings, organ weights and reproduction data:
-The Dunnett-test (many to one t-test) based on a pooled variance estimate was 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) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test was applied to the macroscopic findings.
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
In group 4, all animals were noted to push the head through the bedding material starting on day 9 of the pre-pairing period onwards. One female had salivation after administration of the test item on day 9 of the gestation period. These signs are considered to be signs of discomfort and not to be adverse. In groups 2 and 3, no clinical signs or observations were noted.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One female in group 2 died accidentally after blood sampling. This death was not considered to be test item-related.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- Males - pre-pairing period: In group 4, mean body weight gain was statistically significantly decreased during the prepairing period (+6.6% versus +13.7% in the control group). Although this decrease had no statistically significant impact on the mean body weight, it was considered to be a test item related effect. During the pairing period, mean body weight and mean body weight gain were not affected by treatment with the test item. In groups 2 and 3, mean body weight and mean body weight gain were not affected by the treatment with the test item for the entire duration of the study. In group 3, mean body weight gain was statistically significant lower on day 6 of the pre-pairing period however, statistical significance occurred on a single day only and therefore it was not considered to be an adverse effect. During the pairing period, mean body weight gain was lower (+8.2% versus +12.0% in the control group) attaining statistically significance on days 9, 13 and 14. However, this was considered to be incidental as there was no-dose dependency.
- Females - pre-pairing, gestation and lactation periods: In group 4, mean body weight gain was statistically significant decreased during the prepairing period (+3.5% versus +8.2% in the control group). This was considered to be a test item related effect, although it did not affect the mean body weight. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 3, mean body weight gain was lower during the pre-pairing period (+5.0% versus +8.2% in the control group) attaining statistical significance between day 6 and 8 and on days 11 and 14. This was considered an effect of the test item although mean body weight was not affected. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 2, mean body weight and mean body weight gain were not affected for the entire duration of the treatment with the test item.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- Males - pre-pairing period: In group 4, mean food consumption was statistically significant reduced between days 1-8 of the pre-pairing period (-19.2% compared to the control group) and it remained lower between days 8-14 (-10.6% compared to the control group) without attaining statistical significance. In group 3, mean food consumption was not considered to be affected by the treatment with the test item. As mean food consumption was slightly lower between days 1 and 8 of the pre-pairing period (-4.9% compared to the control group) and slightly higher between days 8 and 14 of the pre-pairing period (+2.1% compared to the control group), these variations were considered to be incidental. In group 2, no test item-related effects were noted.
- Females - pre-pairing, gestation and lactation periods: In group 4, mean food consumption was statistically significant decreased between days 1-8 of the pre-pairing period (-15.6% compared to the control group). This was considered to be a test item-related effect. During the gestation and lactation periods mean food consumption was not considered to be affected by treatment with the test item. The statistically significant higher food consumption during the lactation period (+23.0% compared to the control group) was considered to be incidental. In group 3, mean food consumption was decreased between days 1-8 of the pre-pairing period (-10.8% compared to the control group). This was considered to be a transient test item-related effect even though it was not statistically significant. During the gestation and lactation periods, no test item-related effects were noted. In group 2, mean food consumption was not affected by treatment with the test item. During the lactation period, the statistically significant higher mean food consumption was considered to be incidental.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
- Males: In group 4, the absolute count of neutrophils was statistically significant higher (+40.6% compared to the control group). Since the relative count was not statistically significant increased, this was not considered to be adverse. Mean corpuscular hemoglobin concentration was slightly but statistically significant decreased (-2.8% compared to the control group). Since the mean corpuscular hemoglobin was not affected, it was not considered to be an adverse effect. In group 3, the statistically significant higher relative red cell volume distribution width (+19.0% compared to the control group) was within the range of historical reference value. In group 2 no changes were noted.
- Females: The assessment of the hematology data did not reveal any test item-related effects in females.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
- Males: In group 4, urea and potassium concentrations were statistically significant increased (+25.0% and +15.2%, respectively, compared to the control group). These alterations correlate with the histopathological findings noted in the kidney, thus they were considered to be test item-related. In group 3, the statistically significant lower phosphorus concentration (-13.6% compared to the control group) was within the range of historical reference values. The statistically significant lower concentration of total bilirubin in groups 2 and 3 (-27.6% and -26.8% compared to the control group, respectively) was not considered to be a test item-related effect because there was no dose-dependency.
- Females: The assessment of the clinical biochemistry parameters did not reveal any test item-related effect.
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
- Functional Observational Battery: There was no indication of any test item-related effect during the open field phase. Mean values of grip strength (fore- and hind paws) and landing foot splay gave no indication of any test item-related effects. In groups 3 and 4, mean body temperature was statistically significant reduced in males compared to the control group (37.5°C and 37.4°C, respectively, compared to 38.6 °C in the control group).
- Locomotor Activity: Locomotor activity was assessed quantitatively in terms of low beam counts in an activity monitor. In all groups there was no indication of a test item-related effect. The statistically significant lower locomotor activity noted at 30 min in group 4 males, and the statistically significant lower total locomotor activity noted in group 4 females were within the range of the historical control data.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
- Males: In group 4, mean relative liver and kidney weights were statistically significant increased.
- Females: In group 4, mean absolute and relative liver weights were statistically significant increased.
These findings correlated with the histopathological findings and were therefore considered to be test item-related.
Gross pathological findings:
no effects observed
Description (incidence and severity):
The findings noted during the macroscopic examinations did not give any indication of test item related effects.
- Males: In group 4, one male was noted to have reddish foci on the thymus, a second male was noted to have a dark red discoloration of the mandibular lymph node and a third male to have enlarged liver. Dark red or reddish foci were noted on the thymus of one male in group 3, in three males in group 2 and in one male in group 1. One male in group 1 was noted to have a subcutaneous yellowish-soft nodule on the right side of thoracic dorsal region.
- Females: In group 4, an isolated reddish focus in the lung of one female and dark red foci in the stomach of a second female were noted. In group 3, one female was noted to have a dark brown content in the stomach, jejunum and ileum. Dark red foci were also noted on the mucosa of fundus. The other findings noted were dark red discoloration of both ovaries in a second female and enlargement of the renal lymph nodes in a third female. In group 2, dark red foci were noted on the thymus in two females (one of these females died after the blood sampling).
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
- Liver: Minimal centrilobular hepatocellular hypertrophy was observed in four males in group 3 and slight centrilobular hepatocellular hypertrophy in one male in group 4; minimal to slight diffuse hepatocellular hypertrophy was noted in the remaining four males and all five females in group 4. Both types of liver cell hypertrophy were considered to represent an adaptive response to the increased need for metabolizing the test item.
- Thyroid gland: In group 4, minimally increased incidence and severity of diffuse follicular cell hypertrophy was noted in males and females. This change was considered to be secondary to the enhanced liver cell metabolism due to hepatocellular hypertrophy.
- Stomach: Minimal acanthosis of nonglandular stomach was present in all males and in three females in group 4 and was associated with minimal to slight focal/multifocal chronic inflammation in two of these males. Isolated, slight and multifocal chronic inflammation occurred in two females in group 3 and one male in group 4. Minimal multifocal chronic inflammation in the glandular stomach was noted in one male in group 3, and slight focal chronic inflammation in one female in group 3. The findings in the female were associated with minimal and multifocal congestion. Slight multifocal congestion occurred in two females in group 4, in one female associated with an acute thrombus and in the other with minimal and multifocal chronic inflammation. All of the stomach findings were considered to represent a local irritating effect of the test item.
- Kidneys: Minimally increased incidence of hyaline droplets/granules was observed in males in groups 3 and 4 (five males in each group, compared to three in the control group). These eosinophilic hyaline droplets/granules, which reflected an increased content of α-2μ-globulin, occur within the cytoplasm of proximal convoluted tubules of sexually mature male rats. Because a slightly increased severity was also noted (mean severity of 2.2 compared to 1.0 in the control males), this was considered to be an adverse effect of the test item in group 4 males.
- Spleen: Extramedullary hematopoiesis was minimally increased in severity in females in group 4, possibly secondary to or an adaptation to stress and/or impaired health conditions. In group 4, the minimally increased incidence of thymus atrophy in females was also considered to be secondary to stress rather than to represent a direct effect of the test item.
All other microscopic findings noted in various organs and in all groups examined were considered to be incidental in nature because their morphology, severity, and incidence did not distinguish treated rats from controls.
Histopathological findings: neoplastic:
no effects observed
Other effects:
not specified
Dose descriptor:
NOEL
Effect level:
43 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
behaviour (functional findings)
body weight and weight gain
clinical biochemistry
food consumption and compound intake
histopathology: non-neoplastic
Dose descriptor:
NOAEL
Effect level:
160 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
food consumption and compound intake
histopathology: non-neoplastic
Critical effects observed:
yes
Lowest effective dose / conc.:
600 mg/kg bw/day (actual dose received)
System:
urinary
Organ:
kidney
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
no

Table 1: differences in mean Food consumption (Males in g/animal/day), pre-pairing period

Group

Days

 

1 – 8

8 - 14

1 - 14

mg/kg

g

(%)*

g

(%)*

g

(%)*

1 (0)

24.5

 

23.5

 

24

 

2 (43)

24.3

-0.8

25.6

+8.9

25

+4.2

3 (160)

23.3

-4.9

24

+2.1

23.7

-1.3

4 (600)

19.8

-19.2

21

-10.6

20.4

-15.0

*percentages refer to the values of group1

 

Table 2: Differences in mean Body weight gain males, pre-pairing period

Group

Days

 

1 – 8

8 - 14

1 - 14

mg/kg

g

(%)*

g

(%)*

g

(%)*

1 (0)

22

+7.0

21

+6.3

43

+13.7

2 (43)

20

+6.3

23

+6.8

43

+13.5

3 (160)

16

+5.0

17

+5.1

33

+10.4

4 (600)

10

+3.1

11

+3.3

21

+6.6

*increase within the respective time interval

 

Table 3: differences in mean Food consumption (Females in g/animal/day), pre-pairing period

Group

Days

 

1 – 8

8 - 14

1 - 14

mg/kg

g

(%)*

g

(%)*

g

(%)*

1 (0)

16.7

 

15.3

 

16

 

2 (43)

16.6

-0.6

16.7

+9.2

16.6

+3.8

3 (160)

14.9

-10.8

16.3

+6.5

15.6

-2.5

4 (600)

14.1

-15.6

15.7

+2.6

14.9

-6.9

*percentages refer to the values of group1

 

Table 4: Differences in mean Body weight gain females, pre-pairing period

Group

Days

 

1 – 8

8 - 14

1 - 14

mg/kg

g

(%)*

g

(%)*

g

(%)*

1 (0)

5

+2.6

11

+5.5

16

+8.2

2 (43)

6

+3.0

7

+3.4

13

+6.4

3 (160)

0

+0.0

10

+5.0

10

+5.0

4 (600)

1

+0.5

6

+3.0

7

+3.5

*increase within the respective time interval

 

Table 5: Clinical Biochemistry

 

 

Group 1

Group 2

Group 3

Group 4

Days (pre-pairing period)

 

0

43 mg/kg bw

160 mg/kg bw

600 mg/kg bw

Male (before necropsy)

Urea

MEAN

6.16

5.93

6.79

7.70*

mmol/L

% of control

100

96

110

125

 

SD

0.63

0.93

0.72

1.16

 

N

5

5

5

5

Potassium

MEAN

4.09

4.27

4.34

4.71*

mmol/L

% of control

100

104

106

115

 

SD

0.39

0.24

0.18

0.49

 

N

5

5

5

5

Female (day 5 post partum)

Urea

MEAN             

9.34

8.16

8.03

8.79

mmol/L

% of control

100

87

86

94

 

SD

1.82

1.28

0.63

2.12

 

N

5

5

5

5

Potassium

MEAN

4.10

4.22

3.92

3.77

mmol/L

% of control

100

103

96

92

 

SD

0.59

0.80

0.53

0.50

 

N

5

5

5

5

Significant different compared to control value, *p<0.05

 

Table 6: Organ/body weight ratios

 

 

Group 1

Group 2

Group 3

Group 4

Days (pre-pairing period)

 

0

43 mg/kg bw

160 mg/kg bw

600 mg/kg bw

Male

Liver (%)

MEAN

2.57

2.53

2.63

3.02**

 

SD

0.1

0.07

0.16

0.17

 

N

5

5

5

5

Kidneys (%)

MEAN

0.61

0.61

0.61

0.68**

 

SD

0.02

0.02

0.03

0.02

 

N

5

5

5

5

Female

Liver (%)

MEAN

3.18

3.35

3.19

3.93**

 

SD

0.21

0.07

0.25

0.23

 

N

5

5

5

5

Kidneys (%)

MEAN

0.63

0.65

0.61

0.67

 

SD

0.06

0.02

0.03

0.05

 

N

5

5

5

5

Significant different compared to control value, **p<0.01

Reason / purpose for cross-reference:
reference to same study
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 Apr - 28 Nov 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
adopted 22. March 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EPA OPPTS 870.3650
Version / remarks:
July 2000
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
- Physical state: liquid
- Analytical purity: 87%
- Active content: 30.89%
- Lot/batch No.: 184
- Expiration date of the lot/batch: 30. June 2008
- Storage condition of test material: room temperature (20 ± 5°C)
Species:
rat
Strain:
Wistar
Remarks:
HanRcc: WIST(SPF)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories Ltd., Switzerland
- Age at study initiation: 11 weeks
- Weight at study initiation: mean: males 294 - 344 g; females 176 - 221 g
- Housing: individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding
- Diet: Pelleted standard Kliba Nafag 3433 rat/mouse maintenance diet, ad libitum
- Water: community tap-water, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
highly purified
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The dosage formulations were prepared weekly prior to administration using the test item as supplied by the Sponsor and using a factor of 3.72 taking in account the purity of 87% and the content of the active ingredient of 30.89% (CAS 3655-00-3). Sodium coco β-iminodipropionate (CAS 3655-00-3) was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration. Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: sperm in vaginal smear, copulation plug observed referred to as day 0 post coitum
- After successful mating each pregnant female was caged individually
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Several application formulations were prepared and representative analytical samples were collected and dispatched to the analytical laboratories internally. The test item concentrations were determined by HPLC coupled to an ELSD detector and quantified with the area under the peak.
Duration of treatment / exposure:
Males: minimum of 4 weeks (2 weeks prior to mating and 2 weeks of mating)
Females: approximately 7 weeks (2 weeks prior to mating up to day 4 post partum)
Frequency of treatment:
daily, 7 days/week
Dose / conc.:
43 mg/kg bw/day (actual dose received)
Remarks:
Group 2
Dose / conc.:
160 mg/kg bw/day (actual dose received)
Remarks:
Group 3
Dose / conc.:
600 mg/kg bw/day (actual dose received)
Remarks:
Group 4
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dosage levels were selected based on a previous dose range finding toxicity study in Han Wistar Rats.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS
- Time schedule: daily
- changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern), changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported

BODY WEIGHT
- Time schedule for examinations: daily from treatment start to day of necropsy

FOOD CONSUMPTION AND COMPOUND INTAKE
- was recorded for male animals weekly during pre-pairing, for females weekly in during the pre-pairing period, gestation days 0-7, 7-14 and 14-21 post coitum, and days 1-4 post partum; food consumption was not recorded during the pairing period
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, weight gain, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for any gross anomalies
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals, after at least 28 days of dosing
- Maternal animals: All surviving animals, on day 5 post partum

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
For the parent animals, special attention was directed at the organs of the reproductive system. The number of implantation sites and corpora lutea was recorded for all dams with litters. The uteri of non-pregnant females were placed in a solution of ammonium sulfide to visualize possible hemorrhagic areas of implantation sites
- Tissues from all parental males (preserved in neutral phosphate buffered 4% formaldehyde solution): Prostate, Seminal vesicles with coagulating gland, Testes (in Bouin’s fixative), Epididymides (in Bouin’s fixative)
- Tissues from all parental females (preserved in neutral phosphate buffered 4% formaldehyde solution): Ovaries
- Tissues (preserved in neutral phosphate buffered 4% formaldehyde solution) from the five males and females per group selected for organ weights: Gross lesions, Brain (cerebrum, cerebellum, pons), Spinal chord, Small and large intestines (incl. Peyer’s patches), Stomach, Liver, Kidneys, Adrenals, Spleen, Heart, Thymus, Thyroid and Parathyroid, Trachea and lungs (preserved by inflation with fixative and then immersion), Uterus (with vagina), Urinary bladder, Lymph nodes (mandibular and mesenteric), Peripheral nerve (sciatic), Bone marrow.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring was sacrificed at day 5 post partum
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows: examined macroscopically for any structural changes

Statistics:
The following statistical methods may be used to food consumption, body weight, macroscopic findings, organ weights and reproduction data:
- The Dunnett-test (many to one t-test) based on a pooled variance estimate was 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) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test was applied to the macroscopic findings.
Offspring viability indices:
Birth index = number of pups born alive/number of implantations
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
In group 4, all animals were noted to push the head through the bedding material starting on day 9 of the pre-pairing period onwards. One female had salivation after administration of the test item on day 9 of the gestation period. These signs are considered to be signs of discomfort and not to be adverse. In groups 2 and 3, no clinical signs or observations were noted.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One female in group 2 died accidentally after blood sampling. This death was not considered to be test item-related.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- Males - pre-pairing period: In group 4, mean body weight gain was statistically significany decreased during the pre-pairing period (+6.6% versus +13.7% in the control group). Although this decrease had no statistically significant impact on the mean body weight, it was considered to be a test item related effect. During the pairing period, mean body weight and mean body weight gain were not affected by treatment with the test item. In groups 2 and 3, mean body weight and mean body weight gain were not affected by the treatment with the test item for the entire duration of the study. In group 3, mean body weight gain was statistically significant lower on day 6 of the pre-pairing period however, statistical significance occurred on a single day only and therefore it was not considered to be an adverse effect. During the pairing period, mean body weight gain was lower (+8.2% versus +12.0% in the control group) attaining statistically significance on days 9, 13 and 14. However, this was considered to be incidental as there was no-dose dependency.
- Females - pre-pairing, gestation and lactation periods: In group 4, mean body weight gain was statistically significant decreased during the pre-pairing period (+3.5% versus +8.2% in the control group). This was considered to be a test item related effect, although it did not affect the mean body weight. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 3, mean body weight gain was lower during the pre-pairing period (+5.0% versus +8.2% in the control group) attaining statistical significance between day 6 and 8 and on days 11 and 14. This was considered an effect of the test item although mean body weight was not affected. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 2, mean body weight and mean body weight gain were not affected for the entire duration of the treatment with the test item.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- Males - pre-pairing period: In group 4, mean food consumption was statistically significant reduced between days 1-8 of the pre-pairing period (-19.2% compared to the control group) and it remained lower between days 8-14 (-10.6% compared to the control group) without attaining statistical significance. In group 3, mean food consumption was not considered to be affected by the treatment with the test item. As mean food consumption was slightly lower between days 1 and 8 of the pre-pairing period (-4.9% compared to the control group) and slightly higher between days 8 and 14 of the pre-pairing period (+2.1% compared to the control group), these variations were considered to be incidental. In group 2, no test item-related effects were noted.
- Females - pre-pairing, gestation and lactation periods: In group 4, mean food consumption was statistically significant decreased between days 1-8 of the pre-pairing period (-15.6% compared to the control group). This was considered to be a test item-related effect. During the gestation and lactation periods mean food consumption was not considered to be affected by treatment with the test item. The statistically significant higher food consumption during the lactation period (+23.0% compared to the control group) was considered to be incidental. In group 3, mean food consumption was decreased between days 1-8 of the pre-pairing period (-10.8% compared to the control group). This was considered to be a transient test item-related effect even thought it was not statistically significant. During the gestation and lactation periods, no test item-related effects were noted. In group 2, mean food consumption was not affected by treatment with the test item. During the lactation period, the statistically significant higher mean food consumption was considered to be incidental.
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
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Histopathological evaluation of the reproductive organs did not reveal any relevant changes in high-dose animals. Special emphasis on stages of spermatogenesis and histopathology of interstitial testicular cell structure did not reveal any differences between control (group 1) and high-dose (group 4) males
Histopathological findings: neoplastic:
no effects observed
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
effects observed, non-treatment-related
Description (incidence and severity):
All females mated within the first pairing period. The median and mean precoital times were unaffected by treatment with the test item. Mean precoital times were 4.6, 3.5, 5.3 and 4.2 days in groups 1, 2, 3 and 4, respectively. The median precoital time was 3, 3, 6 and 3 days in order of ascending dose level. The higher median precoital time noted in group 3 was considered to be incidental since there was no dose dependency.
The mean duration of gestation was unaffected by exposure to the test item. Mean duration of gestation was 21.3, 21.6, 20.7 and 21.3 days, in order of ascending dose level.
The mean number of implantations per litter and mean incidence of post-implantation loss as a percentage of total implantations were not affected by the treatment with the test item. Mean number of implantation sites was 12.9, 13.8, 13.6 and 13.4 and mean incidence of postimplantation loss was 12.1, 10.9, 7.4 and 6.6% in groups 1, 2, 3 and 4, respectively.
Dose descriptor:
NOEL
Effect level:
600 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects on reproductive parameters observed
Critical effects observed:
no
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Description (incidence and severity):
The birth index (number of pups born alive/number of implantations) was not affected by treatment with the test item. The birth index was 87.9, 89.1, 92.6 and 93.4% in groups 1, 2, 3 and 4, respectively. Mean litter size at first litter check was 11.3 12.3, 12.6 and 12.6 in groups 1, 2, 3 and 4, respectively. Mean postnatal loss was 0.1, 0.0, 0.2 and 0.0 in groups 1, 2, 3 and 4, respectively. Correspondingly, the viability index was 99.0, 100.0, 98.2 and 100.0%.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Mean pup weights on day 1 post partum and during the lactation period (to day 4 post partum) were unaffected by treatment with the test item. On day 1 post partum mean pup weights were 5.9, 6.2, 5.9 and 5.7 g in order of ascending dose level. Mean pup weights on day 4 post partum were 8.4, 8.9, 8.2 and 8.0 g in order of ascending dose level.
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:
effects observed, non-treatment-related
Description (incidence and severity):
Sex ratios at first litter check and on day 4 post partum were unaffected by exposure to the test item. In group 3, the statistically significant lower number of females was considered to be incidental. The proportion of males was 45, 44, 58 and 50%, in order of ascending dose level.
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No abnormal findings were noted at macroscopic examination of the pups.
Histopathological findings:
no effects observed
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Dose descriptor:
NOEL
Generation:
F1
Effect level:
600 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed
Critical effects observed:
no
Reproductive effects observed:
no
Conclusions:
In absence of any altered parameters, the NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 600 mg/kg/day, the highest dose tested.

Data source

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

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)
Version / remarks:
adopted 22. March 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EPA OPPTS 870.3650
Version / remarks:
July 2000
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Disodium N-(2-carboxyethyl)-N-dodecyl-β-alaninate
EC Number:
222-899-0
EC Name:
Disodium N-(2-carboxyethyl)-N-dodecyl-β-alaninate
Cas Number:
3655-00-3
Molecular formula:
C18H35NO4.2Na
IUPAC Name:
disodium N-(2-carboxyethyl)-N-dodecyl-beta-alaninate
Specific details on test material used for the study:
- Physical state: liquid
- Analytical purity: 87%
- Active content: 30.89%
- Lot/batch No.: 184
- Expiration date of the lot/batch: 30. June 2008
- Storage condition of test material: room temperature (20 ± 5°C)

Test animals

Species:
rat
Strain:
Wistar
Remarks:
HanRcc: WIST(SPF)
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Sex: male / female
- Source: Harlan Laboratories Ltd., Switzerland
- Age at study initiation: 11 weeks
- Weight at study initiation: mean: males 294 - 344 g; females 176 - 221 g
- Housing: individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding
- Diet: Pelleted standard Kliba Nafag 3433 rat/mouse maintenance diet, ad libitum
- Water: community tap-water, ad libitum
- Acclimation period: 7 days

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

Administration / exposure

Route of administration:
oral: gavage
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The dosage formulations were prepared weekly prior to administration using the test item as supplied by the Sponsor and using a factor of 3.72 taking in account the purity of 87% and the content of the active ingredient of 30.89% (CAS 3655-00-3). Sodium coco β-iminodipropionate (CAS 3655-00-3) was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration. Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Several application formulations were prepared and representative analytical samples were collected and dispatched to the analytical laboratories internally. The test item concentrations were determined by HPLC coupled to an ELSD detector and quantified with the area under the peak.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: sperm in vaginal smear, copulation plug observed referred to as day 0 post coitum
- After successful mating each pregnant female was caged individually
Duration of treatment / exposure:
Males: minimum of 4 weeks (2 weeks prior to mating and 2 weeks of mating)
Females: approximately 7 weeks (2 weeks prior to mating up to day 4 post partum)
Frequency of treatment:
daily, 7 days/week
Doses / concentrationsopen allclose all
Dose / conc.:
43 mg/kg bw/day (actual dose received)
Remarks:
Group 2
Dose / conc.:
160 mg/kg bw/day (actual dose received)
Remarks:
Group 3
Dose / conc.:
600 mg/kg bw/day (actual dose received)
Remarks:
Group 4
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dosage levels were selected based on a previous dose range finding toxicity study in Han Wistar Rats.

Examinations

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

DETAILED CLINICAL OBSERVATIONS
- Time schedule: daily
- changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern), changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported

BODY WEIGHT
- Time schedule for examinations: daily from treatment start to day of necropsy

FOOD CONSUMPTION AND COMPOUND INTAKE
- was recorded for females weekly in during the pre-pairing period, gestation days 0-7, 7-14 and 14-21 post coitum, and days 1-4 post partum; food consumption was not recorded during the pairing period

SACRIFICE
All surviving animals, on day 5 post partum

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
For the parent animals, special attention was directed at the organs of the reproductive system. The number of implantation sites and corpora lutea was recorded for all dams with litters. The uteri of non-pregnant females were placed in a solution of ammonium sulfide to visualize possible hemorrhagic areas of implantation sites
- Tissues from all parental females (preserved in neutral phosphate buffered 4% formaldehyde solution): Ovaries
- Tissues (preserved in neutral phosphate buffered 4% formaldehyde solution) from the five females per group selected for organ weights: Gross lesions, Brain (cerebrum, cerebellum, pons), Spinal chord, Small and large intestines (incl. Peyer’s patches), Stomach, Liver, Kidneys, Adrenals, Spleen, Heart, Thymus, Thyroid and Parathyroid, Trachea and lungs (preserved by inflation with fixative and then immersion), Uterus (with vagina), Urinary bladder, Lymph nodes (mandibular and mesenteric), Peripheral nerve (sciatic), Bone marrow.
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: No data
- Number of late resorptions: No data
Fetal examinations:
SACRIFICE
- The F1 offspring was sacrificed at day 5 post partum
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows: examined macroscopically for any structural changes

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, weight gain, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for any gross anomalies
Statistics:
The following statistical methods may be used to food consumption, body weight, macroscopic findings, organ weights and reproduction data:
- The Dunnett-test (many to one t-test) based on a pooled variance estimate was 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) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test was applied to the macroscopic findings.
Indices:
Birth index = number of pups born alive/number of implantations

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
In group 4, all animals were noted to push the head through the bedding material starting on day 9 of the pre-pairing period onwards. One female had salivation after administration of the test item on day 9 of the gestation period. These signs are considered to be signs of discomfort and not to be adverse. In groups 2 and 3, no clinical signs or observations were noted.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One female in group 2 died accidentally after blood sampling. This death was not considered to be test item-related.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- Females - pre-pairing, gestation and lactation periods: In group 4, mean body weight gain was statistically significant decreased during the pre-pairing period (+3.5% versus +8.2% in the control group). This was considered to be a test item related effect, although it did not affect the mean body weight. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 3, mean body weight gain was lower during the pre-pairing period (+5.0% versus +8.2% in the control group) attaining statistical significance between day 6 and 8 and on days 11 and 14. This was considered an effect of the test item although mean body weight was not affected. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 2, mean body weight and mean body weight gain were not affected for the entire duration of the treatment with the test item.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- Females - pre-pairing, gestation and lactation periods: In group 4, mean food consumption was statistically significant decreased between days 1-8 of the pre-pairing period (-15.6% compared to the control group). This was considered to be a test item-related effect. During the gestation and lactation periods mean food consumption was not considered to be affected by treatment with the test item. The statistically significant higher food consumption during the lactation period (+23.0% compared to the control group) was considered to be incidental. In group 3, mean food consumption was decreased between days 1-8 of the pre-pairing period (-10.8% compared to the control group). This was considered to be a transient test item-related effect even thought it was not statistically significant. During the gestation and lactation periods, no test item-related effects were noted. In group 2, mean food consumption was not affected by treatment with the test item. During the lactation period, the statistically significant higher mean food consumption was considered to be incidental.
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):
There were no changes in organ weights for organs of the reproductive system.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No gross anomalies were detected.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Histopathological evaluation of the reproductive organs did not reveal any relevant changes in high-dose animals.
Histopathological findings: neoplastic:
no effects observed

Maternal developmental toxicity

Number of abortions:
not examined
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
The mean number of implantations per litter and mean incidence of post-implantation loss as a percentage of total implantations were not affected by the treatment with the test item. Mean number of implantation sites was 12.9, 13.8, 13.6 and 13.4 and mean incidence of postimplantation loss was 12.1, 10.9, 7.4 and 6.6% in groups 1, 2, 3 and 4, respectively.
Total litter losses by resorption:
not examined
Early or late resorptions:
not examined
Dead fetuses:
no effects observed
Changes in pregnancy duration:
effects observed, non-treatment-related
Description (incidence and severity):
The mean duration of gestation was unaffected by exposure to the test item. Mean duration of gestation was 21.3, 21.6, 20.7 and 21.3 days, in order of ascending dose level.
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): effects observed, non-treatment-related
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.DescriptionIncidenceAndSeverityEffectsOnPregnancyDuration): No effects on pregnancy were observed
Changes in number of pregnant:
not examined
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
All females mated within the first pairing period. The median and mean precoital times were unaffected by treatment with the test item. Mean precoital times were 4.6, 3.5, 5.3 and 4.2 days in groups 1, 2, 3 and 4, respectively. The median precoital time was 3, 3, 6 and 3 days in order of ascending dose level. The higher median precoital time noted in group 3 was considered to be incidental since there was no dose dependency.

Effect levels (maternal animals)

Dose descriptor:
NOEL
Effect level:
600 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: no effects on reproductive parameters

Maternal abnormalities

Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
Mean pup weights on day 1 post partum and during the lactation period (to day 4 post partum) were unaffected by treatment with the test item. On day 1 post partum mean pup weights were 5.9, 6.2, 5.9 and 5.7 g in order of ascending dose level. Mean pup weights on day 4 post partum were 8.4, 8.9, 8.2 and 8.0 g in order of ascending dose level.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
The birth index (number of pups born alive/number of implantations) was not affected by treatment with the test item. The birth index was 87.9, 89.1, 92.6 and 93.4% in groups 1, 2, 3 and 4, respectively. Mean litter size at first litter check was 11.3 12.3, 12.6 and 12.6 in groups 1, 2, 3 and 4, respectively. Mean postnatal loss was 0.1, 0.0, 0.2 and 0.0 in groups 1, 2, 3 and 4, respectively. Correspondingly, the viability index was 99.0, 100.0, 98.2 and 100.0%.
Changes in sex ratio:
effects observed, non-treatment-related
Description (incidence and severity):
Sex ratios at first litter check and on day 4 post partum were unaffected by exposure to the test item. In group 3, the statistically significant lower number of females was considered to be incidental. The proportion of males was 45, 44, 58 and 50%, in order of ascending dose level.
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
not specified

Effect levels (fetuses)

Dose descriptor:
NOEL
Effect level:
600 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed as far as F1 can be assessed in a screening study

Fetal abnormalities

Abnormalities:
no effects observed

Overall developmental toxicity

Developmental effects observed:
no

Applicant's summary and conclusion