Registration Dossier
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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 239-032-7 | CAS number: 14960-06-6
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Developmental toxicity / teratogenicity
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:
- 2 008
- 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
- 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
Constituent 1
- 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
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.