Registration Dossier
Registration Dossier
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EC number: 422-630-9 | CAS number: 22208-25-9
Endpoint summary
Administrative data
Key value for chemical safety assessment
Effects on fertility
Description of key information
Weight of Evidence approach:
Method similar to OECD 407. The NOAEL was determined to be at 1000 mg/kg bw/day in rats after 14 days of oral exposure. No gross pathological changes or effects in absolute and relative weights were noted in reproductive organs at any dose tested.
Read-across, test method according to OECD 422 (GLP study). Based on read-across approach from experimental data on analogue methyl acetoacetate, the NOEL for repeat dose toxicity, reproduction performance of parents and development of offspring of the test item AA-TMP was estimated to be 1109.2 mg/kg/day.
Link to relevant study records
- Endpoint:
- fertility, other
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 06 January 2023 – 28 January 2023
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- other: OECD Guideline 407
- Deviations:
- yes
- Remarks:
- (The study period was only 14 days as it was conducted as a dose range finding study for a subsequent more extensive toxicity study)
- GLP compliance:
- no
- Remarks:
- DRF study performed without GLP
- Limit test:
- no
- Species:
- rat
- Strain:
- Sprague-Dawley
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: In-house bred animals.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9 weeks (at receipt)
- Weight at study initiation: 213.78-217.06 g (range of average values of each group of males); 179.99-183.89 g (range of average values of each group of females)
- Fasting period before study: No
- Housing: Maximum of three animals of same sex were housed in a standard polycarbonate cage (Size: L 43 X B 28 X H 21 cm) with stainless steel mesh top grill having facilities for holding pelleted food and drinking water in water bottle fitted with stainless steel sipper tube. Clean sterilized corn cob was provided as bedding material.
- Diet (e.g. ad libitum): Altromin maintenance diet for rats and mice (manufactured by Altromin Spezialfutter GmbH & Co. KG) was provided ad libitum to the animals throughout the acclimatization and experimental period.
- Water (e.g. ad libitum): Water was provided ad libitum throughout the acclimatization and experimental period. Deep bore-well water passed through a Reverse Osmosis Unit was provided in plastic water bottles with stainless-steel sipper tubes.
- Acclimation period: 8 days
DETAILS OF FOOD AND WATER QUALITY:
Analyses of feed and water were performed before starting the study. The corresponding certificates are annexed to the test report.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.1-22.9 ºC
- Humidity (%): 46-66 %
- Air changes (per hr): 12-15
- Photoperiod: 12 hrs dark / 12 hrs light
IN-LIFE DATES: From: 06 January 2023 To: 28 January 2023 - Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
The test item formulations were freshly prepared before dose administration and homogeneity was achieved by thorough stirring using a magnetic stirrer during administration. The required quantity of test item was weighed separately and transferred, mixed and stirred using glass rod and thereafter the entire quantity of the formulation was transferred into measuring cylinder. The rinsing procedure was repeated (many times). Finally, the volume was adjusted to required mark in measuring cylinder with vehicle to get a desired concentration.
VEHICLE
- Justification for use and choice of vehicle (if other than water): In a solubility test, the test item formed suspension in corn oil. Hence, corn oil was selected as vehicle to prepare test item formulations in this study. Corn oil is one of the commonly used vehicles in toxicology studies.
- Concentration in vehicle: 10, 30 and 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Lot/batch no. (if required): not reported.
- Purity: N/A - Analytical verification of doses or concentrations:
- no
- Details on analytical verification of doses or concentrations:
- The stability and homogeneity of the test item in dose formulations was not established under this dose range finding study. However, freshly prepared test item formulations were administered to the animals and homogeneity was achieved by thorough stirring using magnetic stirrer. The actual dose volume for each animal was calculated based on the most recent body weight.
- Duration of treatment / exposure:
- 14 days
- Frequency of treatment:
- Once a day
- Dose / conc.:
- 0 mg/kg bw/day (nominal)
- Remarks:
- G1 - Vehicle control
- Dose / conc.:
- 100 mg/kg bw/day (nominal)
- Remarks:
- G2 - Low dose
- Dose / conc.:
- 300 mg/kg bw/day (nominal)
- Remarks:
- G3 - Mid dose
- Dose / conc.:
- 1 000 mg/kg bw/day (nominal)
- Remarks:
- G4 - High dose
- No. of animals per sex per dose:
- 3
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale:
The doses of 0, 100, 300 and 1000 mg/kg bw/day have been selected for dose range finding study as no literature is available for the test item.
- Fasting period before blood sampling for clinical biochemistry: The animals were fasted overnight before blood collection. Water was provided ad libitum during fasting period. - Positive control:
- None
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily for clinical signs of toxicity and twice daily for mortality and morbidity.
DETAILED CLINICAL OBSERVATIONS: Yes
BODY WEIGHT: Yes
- Time schedule for examinations: Individual animal body weight was recorded on the day of randomization, on day of treatment (Day 1) prior to treatment and weekly thereafter.
FOOD CONSUMPTION: yes
-feed consumption was measured at weekly intervals. Average feed intake per rat (g/rat/day) was calculated using the amount of feed given and left over in each cage and the number of rats surviving in each cage.
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
- Time schedule for collection of blood: On the day of necropsy (day 15)
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes (water provided ad libitum)
- How many animals: all animals
- Parameters checked: Haemoglobin concentration (HGB), Haematocrit (HCT), Erythrocyte count (RBC), Total leukocyte count (WBC), Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular haemoglobin concentration (MCHC), Platelet count (PLT), Mean platelet volume (MPV), Reticulocyte count (Retic), Absolute reticulocyte count, Differential leucocytes count (DLC), Absolute differential leucocytes count (DLC). Prothrombin Time (PT) and Activated Partial Thromboplastin Time (APTT) were estimated by coagulation analyzer.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on the day of necropsy (day 15)
- Animals fasted: Yes (water provided ad libitum)
- How many animals: all animals
- Parameters checked: Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Alkaline phosphatase (ALP), Cholinesterase, Total Protein, Albumin, Total bilirubin, Glucose, Total Cholesterol, Creatinine, Urea, Triglycerides, Phosphorous, Calcium, Blood Urea Nitrogen, Globulin, Albumin/Globulin ratio. Sodium (mmol/L), Potassium (mmol/L) and Chloride (mmol/L) were estimated using Prolyte Na/K/Cl analyzer (Medica Corporation).
URINALYSIS: Yes
- Time schedule for collection of urine: on the day of necropsy (day 15)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked: Blood, Bilirubin, Urobilinogen, Ketones, Protein, Glucose, Leucocytes. In addition pH, nitrite and specific gravity were also analyzed. After analysis of the above parameters, the urine was subjected for centrifugation at 1500 rpm for 3 minutes. Then the urine was subjected for microscopic examination for urine sediments. - Sperm parameters (parental animals):
- Testes and epididymides were subjected to necropsy and detailed gross pathological examination. These organs were weighed and weight ratios as percentage of body weight were determined.
- Postmortem examinations (parental animals):
- SACRIFICE
-On day15, all animals of Group G1, G2, G3 and G4 were subjected to necropsy and detailed gross pathological examination
GROSS NECROPSY
-Gross pathological examination of external surfaces, external orifices, abdominal, thoracic and cranial cavities, as well as organs and tissues of each animal with special emphasis on reproductive organs.
The following organs from all animals at the scheduled sacrifices were weighed wet as soon as possible to avoid drying: Kidneys, Adrenals, Spleen, Heart, Liver, Thymus, Brain, Lungs Testes/Ovaries, Epididymides/Uterus, Prostate along with seminal vesicles with coagulating gland.
HISTOPATHOLOGY / ORGAN WEIGHTS
-The above mentioned organs from all animals were collected, weighed and preserved. The organ weight ratios as percentage of body weight were determined. - Statistics:
- After verification, the data was subjected to statistical analysis using SPSS software, version 27. Body weight, percent change in body weight with respect to day 1, haematological, clinical chemistry estimations parameters absolute and relative organ weights were subjected to statistical analysis. One way ANOVA followed by Dunnett’s post test was done for different treatment groups comparing with the control group data. All analysis and comparisons were evaluated at the 95% level of confidence (P<0.05).
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- No clinical signs of toxicity were seen in all the tested dose group animals in either sex.
- Dermal irritation (if dermal study):
- not examined
- Mortality:
- no mortality observed
- Description (incidence):
- There were no mortality/morbidity observed at any dose group during the experimental period.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- No test item related variations in mean body weight and percent body weight change were noted, when compared to vehicle control group.
- Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- There was not any impact of test item on feed consumption in any of the tested group in either sex.
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no adverse test item related changes in the haematology and coagulation parameters. However, in males statistically significant (p<0.05) increases in Erythrocyte count (RBC), Haemoglobin concentration (HGB), Haematocrit (HCT) Eosinophils in G4 was noted and in females decrease in Prothrombin Time (PT) in G2 and G4 was noted. The observed variation is considered incidental in the absence of dose responsiveness and similar changes were not noted in other sex.
- Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No test item related adverse changes were observed in any of the clinical chemistry parameters, in both sexes when compared to the vehicle control.
However, in G2 female statistically significant (p<0.05) increase in glucose was noted. The noted variation lacked dose responsiveness, hence is considered incidental. - Endocrine findings:
- not examined
- Urinalysis findings:
- no effects observed
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Histopathological findings: non-neoplastic:
- not examined
- Histopathological findings: neoplastic:
- not examined
- Other effects:
- no effects observed
- Reproductive function: oestrous cycle:
- not examined
- Reproductive function: sperm measures:
- no effects observed
- Description (incidence and severity):
- No gross pathological changes or effects in absolute and relative weights were noted in testes or epididymides at any dose tested.
- Reproductive performance:
- not examined
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 1 000 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No treatment related effects found up to highest tested dose of 1000 mg/kg bw/d.
- Key result
- Critical effects observed:
- no
- Remarks on result:
- other: F1 was not assessed as this study was conducted as a dose range finding study for a subsequent more extensive toxicity study
- Key result
- Reproductive effects observed:
- no
- Conclusions:
- The NOAEL was determined to be at 1000 mg/kg bw/day in rats after 14 days of oral exposure.
- Executive summary:
A dose range finding study was performed for the test substance in accordance with OECD guideline 407 in order to evaluate its toxic potential after repeated exposure and select the appropriate doses for a subsequent toxicity study. The test item was diluted in corn oil (vehicle) and added to Sprague-Dawley rats (3 per sex per dose) at doses of 0 (vehicle only), 100, 300 and 1000 mg/kg bw/day for a period of 14 days. The vehicle and test item formulations were administered orally (gavage) at a dose volume of 10 mL/kg body weight. Freshly prepared test item formulations were administered to the animals as soon as possible after preparation.
All animals for each group were observed for clinical signs of toxicity once daily, mortality and morbidity twice daily, and body weight and feed consumption weekly. Clinical pathology (haematology, clinical chemistry analysis and urinalysis) was conducted for all the animals sacrificed at termination. All animals were subjected to gross pathological examination and the organs were weighed, collected and preserved.
No clinical signs of toxicity or mortality were observed. No treatment related changes in mean body weight and percent change in body weight (%) with respect to day 1 and feed consumption were observed. No treatment related changes were observed in haematology, coagulation, clinical chemistry and urinalysis parameters. No treatment related changes in fasting body weights, organ weights and its ratios were noted. No gross pathological changes were observed. Specifically, no gross pathological changes or effects in absolute and relative weights were noted in reproductive organs at any dose tested.
Based on these results, the NOAEL was determined to be at 1000 mg/kg bw/day.
- Endpoint:
- screening for reproductive / developmental toxicity
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
The analogue substance methyl 3-oxobutanoate (methyl acetoacetate) which shares the same functional groups with the substance 1,1,1-Tri((1,3-dioxobutoxy)-methyl)-propane (AA-TMP) also has comparable values for the relevant molecular properties.
See attached the reporting format. - Reason / purpose for cross-reference:
- read-across source
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- No death occurred in any group and no abnormal clinical signs were observed in males. One of female in the 300 mg/kg group showed hypoactivity, bradypnea, hypothermia and prone position from before administration on day 2 of lactation, and died on day 3 of lactation. No death occurred in any female nor were there any abnormal clinical signs observed in the 100 and 1000 mg/kg groups.
- Dermal irritation (if dermal study):
- not examined
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- No death occurred in any group in males. One of female in the 300 mg/kg group showed hypoactivity, bradypnea, hypothermia and prone position from before administration on day 2 of lactation, and died on day 3 of lactation. No death occurred in any female in the 100 and 1000 mg/kg groups.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- No differences were seen in any group as compared to the control group throughout the administration period for males or females.
- Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- No differences were seen in any group as compared to the control group throughout the administration period for males or females.
- 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):
- No differences were seen in any group as compared to the control group for males or females.
- Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- A decrease in T. protein was observed in males of 100 mg/kg group and a decrease in γ-GTP was observed in males in the 300 mg/kg group.
- Endocrine findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, non-treatment-related
- Histopathological findings: non-neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- In males, lymphocytic infiltration was noted in the left ventricular endocardium of 1 animal in the 1000 mg/kg group. Moreover, the effect seen in one animal in the 100 mg/kg group, in which hypertrophy of the spleen was noted macroscopically, was judged to be myeloid leukemia based on the following histopathological findings: there was a depletion of bone marrow cells with normal cytoplasm and filled dominantly with tumor cells with relatively light and circular to elliptic nuclei, and similar tumor cells were also observed in the periportal region and diffusely proliferated in sinusoid in the liver. In the spleen, infiltrative hyperplasia of the tumor cells displaced the normal structure entirely. No tumorous change was noted in the mesenteric lymph nodes. No changes were seen in organs of females in any group. In addition, focal necrosis in the liver, thrombus in the lung, necrosis in the proximal tubular epithelium, hemorrhage in the renal tubule, ulcer in the fore stomach, erosion in the glandular stomach, atrophy in the thymus and hypertrophy in the zona glomerulosa and zona fasciculata were observed in the one animal that died. Furthermore, pulmonary thrombosis was positive by the PTAH staining.
- Histopathological findings: neoplastic:
- no effects observed
- Other effects:
- no effects observed
- Reproductive function: oestrous cycle:
- no effects observed
- Description (incidence and severity):
- No differences were seen in the count of estrous or estrous cycle in any group as compared to the control group.
- Reproductive function: sperm measures:
- not specified
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- In the fertility, copulation was confirmed in all pairs except for 1 pair of the 100 mg/kg group and fertility was confirmed in all pairs. Accordingly, the copulation index was 100%, 91.67%, 100% and 100% in the control, 100, 300 and 1000 mg/kg groups, respectively, no differences were seen in any group as compared to the control group. No differences in the days required for successful copulation were seen in any group as compared to the control group, respectively. Moreover, no abnormality was seen at the necropsy of female which not copulated or of the ovary at the histopathology.
At delivery, the number of newborn females was higher than males, indicating female-biased sex ratio in the 100 mg/kg group, and increase in numbers of implantation, newborns and live newborns was observed in the 300 mg/kg group. However, no differences were seen in any group as compared to the control group in the gestation period, number of corpora lutea, birth and stillbirth indices, numbers of live newborns and body weight of live newborns, and no abnormality was seen by external examination of live newborns in any group. - Key result
- Dose descriptor:
- NOEL
- Remarks:
- (parental toxicity)
- Effect level:
- 1 109.2 mg/kg bw/day (nominal)
- Based on:
- other: Analogue substance
- Sex:
- male/female
- Basis for effect level:
- other: no effects up to the highest tested concentration
- Remarks on result:
- other: Based on read-across approach from experimental data on analogue methyl acetoacetate
- Key result
- Dose descriptor:
- NOEL
- Remarks:
- (reproductive)
- Effect level:
- 1 109.2 mg/kg bw/day (nominal)
- Based on:
- other: Analogue substance
- Sex:
- male
- Basis for effect level:
- other: no effects up to the highest tested concentration
- Remarks on result:
- other: Based on read-across approach from experimental data on analogue methyl acetoacetate
- Key result
- Critical effects observed:
- no
- Clinical signs:
- no effects observed
- Dermal irritation (if dermal study):
- not examined
- Mortality / viability:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- No differences were seen in any group as compared to the control group.
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Sexual maturation:
- not examined
- Anogenital distance (AGD):
- not examined
- Nipple retention in male pups:
- not examined
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- No abnormality was seen by external examination of live newborns in any group.
- Histopathological findings:
- not examined
- Other effects:
- no effects observed
- Behaviour (functional findings):
- not examined
- Developmental immunotoxicity:
- not examined
- Key result
- Dose descriptor:
- NOEL
- Generation:
- F1
- Effect level:
- 1 109.2 mg/kg bw/day (nominal)
- Based on:
- other: Analogue substance
- Sex:
- male/female
- Basis for effect level:
- other: no effects up to the highest tested concentration
- Remarks on result:
- other: Based on read-across approach from experimental data on analogue methyl acetoacetate
- Key result
- Critical effects observed:
- no
- Key result
- Reproductive effects observed:
- no
- Conclusions:
- Based on the read-across approach from experimental results on analogue methyl acetoacetate, the NOEL for repeat dose toxicity, reproduction performance of parents and development of offspring of the test item AA-TMP was estimated to be 1109.2 mg/kg/day.
- Executive summary:
The study was performed in Japan according to OECD-guideline no. 422 as GLP-study. MAA at dosages of 0 (vehicle control), 100, 300 and 1000 mg/kg/day was orally administered to Crj:CD(SD) male and female rats (12 animals/sex/group) from 14 days before mating through the mating period and up to the day before necropsy (Total: 49 days) in males and 14 days prior to mating through the gestation period up to day 3 of delivery in females to investigate the effect on the repeat dose and reproductive toxicity for the parent animals and the development for the offspring, and the following results were obtained.
- Repeat dose toxicity:
There were some effects observed on the clinical signs, blood chemistry and some histopathological parameters. These changes were seen in single animals only, without dose dependency. Therefore, these findings were considered to be of incidental nature and not related to treatment with the test article. No effects were reported on the remaining parameters such as body weight, food consumption, hematology.
- Reproductive/developmental toxicity:
As for reproductive performance in parent animal, no effects of administration of test article were observed on the estrus cycle, numbers of corpora lutea or implantations, copulation (mating performance) or fertility indices. Examination at delivery and during the lactation period revealed, no effects of administration of test article were observed on the gestational days, number of litter or live newborns, gestation, birth or stillbirth indices, sex ratio, body weight at birth or on day 4 after birth, viability index on day 4 after birth. No external malformations in live newborns were observed.
As described above, the no observed effect level (NOEL) under this study conditions was assumed to be 1000 mg/kg/day for repeat dose toxicity in both males and females, and also to be 1000 mg/kg/day for reproductive/developmental toxicity in parent animals and offspring.
Based on these results, the read-across approach was applied and the NOEL for the test item AA-TMP is estimated to be 1109.2 mg/kg/day.
Referenceopen allclose all
Table 1. Summary of clinical signs of toxicity, detailed clinical examination and mortality record
Group, Sex & Dose (mg/kg body weight/day) | No. of Animals | Clinical Signs of Toxicity (No of animals exhibiting signs/Total no. of animals) | Mortality No. of Animals) |
G1, M & 0 | 3 | N | 0/3 |
G2, M & 100 | 3 | N | 0/3 |
G3, M & 300 | 3 | N | 0/3 |
G4, M & 1000 | 3 | N | 0/3 |
G1, F & 0 | 3 | N | 0/3 |
G2, F & 100 | 3 | N | 0/3 |
G3, F & 300 | 3 | N | 0/3 |
G4, F & 1000 | 3 | N | 0/3 |
Table 2. Summary of body weight (g) record
Group, Sex & Dose (mg/kg body weight/day) | Body Weight (g) on Days | ||||
1 | 8 | 14 | |||
G1, M & 0 | Mean | 215.52 | 236.13 | 258.22 | |
±SD | 27.54 | 33.11 | 34.05 | ||
n | 3 | 3 | 3 | ||
G2, M & 100 | Mean | 215.55 | 241.01 | 264.64 | |
±SD | 13.93 | 17.21 | 20.88 | ||
n | 3 | 3 | 3 | ||
G3, M & 300 | Mean | 213.78 | 237.88 | 257.62 | |
±SD | 11.77 | 13.98 | 25.67 | ||
n | 3 | 3 | 3 | ||
G4, M & 1000 | Mean | 217.06 | 240.54 | 259.24 | |
±SD | 5.56 | 6.47 | 10.16 | ||
n | 3 | 3 | 3 | ||
G1, F & 0 | Mean | 180.83 | 193.18 | 207.07 | |
±SD | 11.46 | 9.00 | 6.98 | ||
n | 3 | 3 | 3 | ||
G2, F & 100 | Mean | 183.89 | 193.86 | 210.85 | |
±SD | 13.30 | 15.13 | 15.73 | ||
n | 3 | 3 | 3 | ||
G3, F & 300 | Mean | 179.99 | 193.78 | 202.25 | |
±SD | 5.34 | 3.54 | 3.17 | ||
n | 3 | 3 | 3 | ||
G4, F & 1000 | Mean | 183.35 | 197.98 | 211.12 | |
±SD | 5.14 | 2.66 | 9.26 | ||
n | 3 | 3 | 3 | ||
Table 3. Summary of percent change in body weight (%) with respect to day 1 record
Group, Sex & Dose (mg/kg body weight/day) |
| Percent Change in Body Weight (%) during Days | ||
1 to 8 | 1 to 14 | |||
G1, M & 0 | Mean | 9.44 | 19.77 | |
±SD | 2.03 | 0.50 | ||
n | 3 | 3 | ||
G2, M & 100 | Mean | 11.81 | 22.75 | |
±SD | 2.66 | 4.23 | ||
n | 3 | 3 | ||
G3, M & 300 | Mean | 11.30 | 20.53 | |
±SD | 3.72 | 10.27 | ||
n | 3 | 3 | ||
G4, M & 1000 | Mean | 10.86 | 19.52 | |
±SD | 4.18 | 6.62 | ||
n | 3 | 3 | ||
G1, F & 0 | Mean | 6.91 | 14.66 | |
±SD | 2.02 | 3.72 | ||
n | 3 | 3 | ||
G2, F & 100 | Mean | 5.39 | 14.75 | |
±SD | 0.60 | 6.28 | ||
n | 3 | 3 | ||
G3, F & 300 | Mean | 7.75 | 12.47 | |
±SD | 4.86 | 5.07 | ||
n | 3 | 3 | ||
G4, F & 1000 | Mean | 8.02 | 15.18 | |
±SD | 2.45 | 5.08 | ||
n | 3 | 3 | ||
Table 4. Summary of feed consumption (g/rat/day) record
Group, Sex & Dose (mg/kg body weight/day) |
| Week 1 | Week 2 | |
G1, M & 0 | Mean | 19.01 | 19.59 | |
±SD | - | - | ||
n | 1 | 1 | ||
G2, M & 100 | Mean | 19.64 | 22.03 | |
±SD | - | - | ||
n | 1 | 1 | ||
G3, M & 300 | Mean | 20.34 | 20.40 | |
±SD | - | - | ||
n | 1 | 1 | ||
G4, M & 1000 | Mean | 21.02 | 20.49 | |
±SD | - | - | ||
n | 1 | 1 | ||
G1, F & 0 | Mean | 16.99 | 16.83 | |
±SD | - | - | ||
n | 1 | 1 | ||
G2, F & 100 | Mean | 15.12 | 15.99 | |
±SD | - | - | ||
n | 1 | 1 | ||
G3, F & 300 | Mean | 16.82 | 17.44 | |
±SD | - | - | ||
n | 1 | 1 | ||
G4, F & 1000 | Mean | 16.64 | 17.46 | |
±SD | - | - | ||
n | 1 | 1 |
Table 5. Summary of gross pathology findings
Sex | Male | Female | ||||||
Group | G1 | G2 | G3 | G4 | G1 | G2 | G3 | G4 |
Dose (mg/kg body weight/day) | 0 | 100 | 300 | 1000 | 0 | 100 | 300 | 1000 |
Number of animals | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
No. of dead rats during treatment | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
No. of moribund sacrificed rats | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
No. of terminally sacrificed rats | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
No of rats showing gross pathology | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
Table 6. Summary of haematology record
Group, Sex & Dose (mg/kg body weight/day) |
| Total Leucocyte Count | Total Erythrocyte Count | Hemoglobin | Haematocrit | Mean Corpuscular Volume | Mean Corpuscular Hemoglobin | Mean Corpuscular Hemoglobin Concentration |
(WBC) | (RBC) | (HGB) | (HCT) | (MCV) | (MCH) | (MCHC) | ||
(103 cells/µL) | (106 cells/µL) | (g/dL) | (%) | (fL) | (pg) | (g/dL) | ||
G1, M & 0 | Mean | 15.41 | 7.66 | 15.03 | 44.63 | 58.30 | 19.63 | 33.63 |
±SD | 3.40 | 0.18 | 0.58 | 2.12 | 2.84 | 0.95 | 0.49 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, M & 100 | Mean | 20.59 | 7.85 | 15.50 | 47.43 | 60.53 | 19.83 | 32.73 |
±SD | 3.47 | 0.31 | 0.46 | 0.50 | 2.85 | 1.24 | 0.60 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, M & 300 | Mean | 11.31 | 7.88 | 15.53 | 47.27 | 59.93 | 19.70 | 32.87 |
±SD | 3.65 | 0.11 | 0.45 | 2.15 | 2.28 | 0.36 | 0.64 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, M & 1000 | Mean | 13.96 | 8.61* | 16.37* | 49.90* | 57.97 | 19.00 | 32.77 |
±SD | 2.38 | 0.14 | 0.51 | 1.25 | 0.61 | 0.46 | 0.57 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G1, F & 0 | Mean | 14.09 | 7.75 | 14.87 | 43.47 | 56.10 | 19.20 | 34.23 |
±SD | 2.26 | 0.80 | 1.25 | 4.44 | 0.35 | 0.44 | 0.85 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, F & 100 | Mean | 14.20 | 7.76 | 15.57 | 43.93 | 56.73 | 20.20 | 35.63 |
±SD | 5.77 | 0.93 | 0.58 | 4.95 | 3.25 | 1.81 | 2.89 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, F & 300 | Mean | 13.26 | 7.26 | 14.77 | 41.33 | 56.90 | 20.43 | 35.93 |
±SD | 5.42 | 0.75 | 0.59 | 4.48 | 1.11 | 1.36 | 2.55 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, F & 1000 | Mean | 12.50 | 7.65 | 14.93 | 44.07 | 57.63 | 19.47 | 33.83 |
±SD | 1.75 | 0.41 | 0.57 | 2.06 | 1.17 | 0.31 | 0.55 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
Group, Sex & Dose (mg/kg body weight/day) |
| Platelet Count | Mean Platelet Volume | Reticulocyte Count | Neutrophils | Lymphocytes | Monocytes | Eosinophils | Basophils |
(PLT) | (MPV) | (Retic) | (Neut) | (Lymph) | (Mono) | (Eos) | (Baso) | ||
(103 cells/µL) | (fL) | (%) | (%) | (%) | (%) | (%) | (%) | ||
G1, M & 0 | Mean | 713.00 | 7.47 | 4.04 | 15.50 | 78.80 | 3.10 | 1.00 | 0.40 |
±SD | 464.46 | 0.38 | 1.21 | 5.30 | 6.46 | 1.30 | 0.50 | 0.10 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, M & 100 | Mean | 946.00 | 7.53 | 4.37 | 18.50 | 75.77 | 2.43 | 1.17 | 0.47 |
±SD | 239.95 | 0.21 | 0.71 | 14.04 | 14.26 | 0.75 | 1.24 | 0.15 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, M & 300 | Mean | 913.33 | 7.73 | 2.96 | 18.10 | 71.87 | 5.80 | 2.23 | 0.30 |
±SD | 88.64 | 0.21 | 0.61 | 0.60 | 0.38 | 1.31 | 1.02 | 0.00 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, M & 1000 | Mean | 994.67 | 7.53 | 3.23 | 24.10 | 65.77 | 4.53 | 4.00* | 0.33 |
±SD | 104.27 | 0.21 | 0.86 | 2.46 | 2.21 | 1.89 | 1.01 | 0.06 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G1, F & 0 | Mean | 937.00 | 7.60 | 3.42 | 19.10 | 74.00 | 3.77 | 1.20 | 0.33 |
±SD | 307.15 | 0.20 | 2.00 | 3.89 | 2.89 | 1.21 | 0.46 | 0.06 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, F & 100 | Mean | 764.67 | 8.20 | 3.40 | 16.30 | 75.63 | 2.87 | 3.20 | 0.43 |
±SD | 361.51 | 0.50 | 1.52 | 7.47 | 10.83 | 1.65 | 2.31 | 0.06 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, F & 300 | Mean | 896.00 | 7.93 | 4.09 | 19.03 | 72.40 | 3.87 | 2.13 | 0.40 |
±SD | 162.86 | 0.25 | 1.84 | 5.69 | 5.96 | 0.50 | 1.33 | 0.17 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, F & 1000 | Mean | 985.67 | 8.13 | 3.85 | 22.40 | 71.70 | 3.13 | 1.53 | 0.27 |
±SD | 65.52 | 0.15 | 0.28 | 4.26 | 4.79 | 0.75 | 0.59 | 0.06 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
Group, Sex & Dose (mg/kg body weight/day) |
| Absolute Reticulocyte Count | Absolute Neutrophils | Absolute Lymphocytes | Absolute Monocytes | Absolute Eosinophils | Absolute Basophils | Prothrombin Time | Activated Prothrombin Time |
(Retic) | (Neut) | (Lymph) | (Mono) | (Eos) | (Baso) | (PT) | (APTT) | ||
(109 cells/L) | (103 cells/µL) | (103 cells/µL) | (103 cells/µL) | (103 cells/µL) | (103 cells/µL) | (Seconds) | (Seconds) | ||
G1, M & 0 | Mean | 308.97 | 2.30 | 12.24 | 0.48 | 0.14 | 0.06 | 17.33 | 17.37 |
±SD | 90.40 | 0.51 | 3.50 | 0.21 | 0.06 | 0.04 | 1.25 | 0.84 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, M & 100 | Mean | 343.43 | 3.53 | 15.89 | 0.50 | 0.21 | 0.10 | 16.36 | 20.59 |
±SD | 62.23 | 2.11 | 5.20 | 0.15 | 0.21 | 0.04 | 0.57 | 1.98 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, M & 300 | Mean | 233.13 | 2.06 | 8.12 | 0.62 | 0.28 | 0.03 | 16.27 | 18.77 |
±SD | 49.68 | 0.73 | 2.60 | 0.04 | 0.22 | 0.02 | 0.77 | 2.34 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, M & 1000 | Mean | 277.90 | 3.33 | 9.21 | 0.63 | 0.57 | 0.05 | 16.61 | 16.25 |
±SD | 75.02 | 0.32 | 1.76 | 0.30 | 0.23 | 0.01 | 1.36 | 1.57 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G1, F & 0 | Mean | 254.47 | 2.71 | 10.41 | 0.51 | 0.17 | 0.05 | 17.62 | 19.53 |
±SD | 122.89 | 0.86 | 1.54 | 0.13 | 0.05 | 0.02 | 0.61 | 4.33 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, F & 100 | Mean | 261.13 | 2.07 | 11.05 | 0.39 | 0.39 | 0.06 | 15.72* | 17.88 |
±SD | 118.33 | 0.40 | 5.73 | 0.19 | 0.18 | 0.02 | 0.42 | 5.81 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, F & 300 | Mean | 297.23 | 2.39 | 9.70 | 0.53 | 0.28 | 0.06 | 16.50 | 18.70 |
±SD | 145.43 | 0.64 | 4.50 | 0.29 | 0.17 | 0.05 | 0.82 | 3.98 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, F & 1000 | Mean | 294.43 | 2.75 | 9.02 | 0.38 | 0.18 | 0.03 | 16.09* | 21.79 |
±SD | 18.86 | 0.12 | 1.84 | 0.09 | 0.05 | 0.02 | 0.36 | 0.81 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
*: Statistically significant (P<0.05)
Table 7. Summary of clinical chemistry record
Group, Sex & Dose (mg/kg body weight/day) |
| Glucose | Urea | Creatinine | Total Cholesterol | Triglycerides | Total Protein | Albumin | Alanine aminotransferase | Aspartate aminotransferase | Alkaline phosphatase |
(GLU) | (CRE) | (CHO) | (TRI) | (TPR) | (ALB) | (ALT) | (AST) | (ALP) | |||
(mg/dL) | (mg/dL) | (mg/dL) | (mg/dL) | (mg/dL) | g/dL) | (g/dL) | (U/L) | (U/L) | (U/L) | ||
G1, M & 0 | Mean | 101.67 | 29.37 | 0.55 | 58.33 | 66.67 | 7.20 | 3.22 | 53.33 | 121.67 | 221.33 |
±SD | 10.60 | 5.35 | 0.02 | 4.93 | 17.04 | 0.20 | 0.10 | 4.93 | 24.19 | 74.10 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, M & 100 | Mean | 96.33 | 29.53 | 0.57 | 62.00 | 55.33 | 7.50 | 3.30 | 68.33 | 129.67 | 236.33 |
±SD | 7.64 | 5.85 | 0.02 | 12.77 | 13.87 | 0.30 | 0.17 | 3.21 | 7.23 | 54.50 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, M & 300 | Mean | 89.67 | 29.80 | 0.52 | 51.67 | 101.33 | 7.03 | 3.33 | 61.67 | 122.33 | 341.67 |
±SD | 4.16 | 5.20 | 0.01 | 6.43 | 65.96 | 0.21 | 0.16 | 9.07 | 20.55 | 89.40 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, M & 1000 | Mean | 97.00 | 29.00 | 0.54 | 55.67 | 59.33 | 7.33 | 3.45 | 58.33 | 113.33 | 328.33 |
±SD | 4.36 | 5.55 | 0.04 | 8.50 | 35.73 | 0.49 | 0.17 | 9.07 | 6.03 | 32.65 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G1, F & 0 | Mean | 86.33 | 39.23 | 0.57 | 62.00 | 72.67 | 7.67 | 3.38 | 55.00 | 127.00 | 247.00 |
±SD | 4.16 | 6.38 | 0.04 | 7.81 | 33.65 | 0.06 | 0.09 | 9.00 | 17.78 | 90.54 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, F & 100 | Mean | 104.67* | 35.70 | 0.60 | 65.67 | 61.67 | 7.70 | 3.46 | 99.33 | 181.33 | 182.33 |
±SD | 4.51 | 10.36 | 0.03 | 16.44 | 32.13 | 0.35 | 0.24 | 68.07 | 58.14 | 60.34 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, F & 300 | Mean | 95.33 | 29.93 | 0.54 | 54.67 | 75.33 | 7.37 | 3.26 | 57.00 | 139.33 | 232.00 |
±SD | 8.08 | 4.66 | 0.03 | 5.13 | 39.88 | 0.25 | 0.20 | 4.58 | 17.39 | 164.79 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, F & 1000 | Mean | 98.67 | 24.57 | 0.53 | 73.00 | 66.67 | 7.33 | 3.52 | 55.00 | 106.33 | 204.33 |
±SD | 5.86 | 0.90 | 0.01 | 22.65 | 23.86 | 0.50 | 0.22 | 2.65 | 11.02 | 55.37 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
Group, Sex & Dose (mg/kg body weight/day) |
| Total Bilirubin | Calcium | Phosphorous | Cholinesterase | Globulin | Albumin/Globulin ratio | Blood Urea Nitrogen | Sodium | Potassium | Chloride |
(BIT) | (CAL) | (PHO) | (CHE) | (GLO) | (A/G Ratio) | (BUN) | (Na) | (K) | (CLO) | ||
(mg/dL) | (mg/dL) | (mg/dL) | (U/L) | (g/dL) | - | (mg/dL) | (mmol/L) | (mmol/L) | (mmol/L) | ||
G1, M & 0 | Mean | 0.11 | 10.03 | 6.13 | 154.67 | 3.98 | 0.81 | 13.73 | 139.60 | 3.71 | 105.57 |
±SD | 0.02 | 0.12 | 0.60 | 16.17 | 0.27 | 0.08 | 2.50 | 1.71 | 0.37 | 2.20 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, M & 100 | Mean | 0.07 | 10.10 | 6.05 | 121.00 | 4.20 | 0.79 | 13.80 | 141.87 | 3.93 | 107.47 |
±SD | 0.01 | 0.46 | 0.34 | 37.64 | 0.14 | 0.03 | 2.73 | 0.15 | 0.56 | 1.00 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, M & 300 | Mean | 0.11 | 10.13 | 6.12 | 101.00 | 3.71 | 0.90 | 13.92 | 140.10 | 3.65 | 104.30 |
±SD | 0.04 | 0.49 | 0.64 | 42.53 | 0.09 | 0.04 | 2.43 | 0.46 | 0.18 | 4.06 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, M & 1000 | Mean | 0.10 | 10.27 | 5.98 | 115.33 | 3.88 | 0.89 | 13.55 | 139.23 | 3.66 | 102.63 |
±SD | 0.02 | 0.25 | 0.12 | 11.59 | 0.34 | 0.05 | 2.59 | 2.25 | 0.10 | 3.91 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G1, F & 0 | Mean | 0.09 | 10.20 | 5.69 | 448.67 | 4.29 | 0.79 | 18.34 | 139.53 | 3.81 | 106.87 |
±SD | 0.04 | 0.10 | 0.34 | 218.60 | 0.12 | 0.04 | 2.98 | 1.70 | 0.14 | 1.78 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, F & 100 | Mean | 0.06 | 10.03 | 4.96 | 488.33 | 4.24 | 0.82 | 16.68 | 141.23 | 3.73 | 108.33 |
±SD | 0.01 | 0.31 | 1.52 | 401.85 | 0.46 | 0.14 | 4.84 | 1.27 | 0.43 | 1.08 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, F & 300 | Mean | 0.08 | 9.87 | 5.18 | 415.33 | 4.11 | 0.79 | 13.99 | 139.63 | 3.74 | 107.10 |
±SD | 0.02 | 0.15 | 0.31 | 199.14 | 0.07 | 0.04 | 2.18 | 1.25 | 0.07 | 1.75 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, F & 1000 | Mean | 0.08 | 9.97 | 5.28 | 362.67 | 3.81 | 0.93 | 11.48 | 140.93 | 3.72 | 105.77 |
±SD | 0.01 | 0.35 | 0.47 | 208.41 | 0.29 | 0.02 | 0.43 | 0.84 | 0.12 | 2.20 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
*: Statistically significant (P<0.05)
Table 8. Summary of urinalysis record
Examination | Group, Sex & Dose (mg/kg/day) | G1, M & 0 | G2, M & 100 | G3, M & 300 | G4, M & 1000 | |
Number of Animals | 3 | 3 | 3 | 3 | ||
Physical Examination | Color | Pale Yellow | 3 | - | 3 | 3 |
Yellow | - | 3 | - | - | ||
Apperance | Clear | 3 | 1 | 3 | 3 | |
Turbidity | - | 2 | - | - | ||
Volume (mL) | Mean | 2.2 | 2.7 | 1.2 | 3.0 | |
±SD | 0.8 | 0.3 | 0.3 | 0.5 | ||
Chemical Examination | pH | Mean | 5.8 | 6.3 | 5.7 | 5.8 |
±SD | 0.3 | 0.3 | 0.3 | 0.3 | ||
Specific gravity (SG) | Mean | 1.030 | 1.030 | 1.030 | 1.030 | |
±SD | 0.000 | 0.000 | 0.000 | 0.000 | ||
Urobilinogen (UBG) (mg/dL) | Mean | 0.7 | 0.2 | 1.0 | 0.7 | |
±SD | 0.5 | 0.0 | 0.0 | 0.5 | ||
Bilirubin (BIL) (mg/dL) | 1 | 3 | 3 | 3 | 3 | |
Ketones (KET) (mg/dL) | 5 | 3 | 3 | 3 | 2 | |
15 | - | - | - | 1 | ||
Blood (BLD) (Ery/µL) | Neg | 2 | 2 | - | - | |
Ca80 | - | - | - | 1 | ||
Ca10 | - | 1 | 1 | - | ||
Ca25 | - | - | 2 | 2 | ||
>=Ca200 | 1 | - | - | - | ||
Protein (PRO) (mg/dL) | 30 | 3 | 3 | 3 | 3 | |
Nitrite (NIT) | Neg | 2 | - | 1 | 2 | |
Pos | 1 | 3 | 2 | 1 | ||
Leucocytes (LEU) (Leu/µL) | Ca70 | 2 | 3 | - | - | |
Ca15 | 1 | - | 3 | 3 | ||
Glucose (GLU) (mg/dL) | Neg | 3 | 3 | 3 | 3 | |
Microscopic Examination | Epi Cells | 0 | 2 | 1 | 3 | 2 |
0-1 | 1 | 1 | - | 1 | ||
1-2 | - | 1 | - | - | ||
Casts | Absent | 3 | 3 | 3 | 3 | |
Crystals | Present | 3 | 3 | 3 | 3 | |
Examination | Group, Sex & Dose (mg/kg/day) | G1, F & 0 | G2, F & 100 | G3, F & 300 | G4, F & 1000 | |
Number of Animals | 3 | 3 | 3 | 3 | ||
Physical Examination | Color | Pale Yellow | 3 | 3 | 2 | 3 |
Yellow | - | - | 1 | - | ||
Apperance | Clear | 3 | 3 | 2 | 3 | |
Turbidity | - | - | 1 | - | ||
Volume (mL) | Mean | 1.2 | 2.7 | 2.2 | 4.3 | |
±SD | 0.3 | 0.8 | 0.8 | 1.8 | ||
Chemical Examination | pH | Mean | 6.0 | 5.8 | 7.3 | 5.8 |
±SD | 0.0 | 0.3 | 1.6 | 0.3 | ||
Specific gravity (SG) | Mean | 1.030 | 1.030 | 1.017 | 1.025 | |
±SD | 0.000 | 0.000 | 0.012 | 0.005 | ||
Urobilinogen (UBG) (mg/dL) | Mean | 0.7 | 0.2 | 0.7 | 0.2 | |
±SD | 0.5 | 0.0 | 0.5 | 0.0 | ||
Bilirubin (BIL) (mg/dL) | Neg | - | 1 | - | 2 | |
1 | 3 | 2 | 3 | 1 | ||
Ketones (KET) (mg/dL) | 5 | 3 | 2 | 3 | 1 | |
Neg | - | 1 | - | 2 | ||
Blood (BLD) (Ery/µL) | Neg | 3 | 1 | 3 | 3 | |
Ca10 | - | 2 | - | - | ||
Protein (PRO) (mg/dL) | Neg | - | - | - | 3 | |
Trace | - | 3 | 1 | - | ||
>=300 | - | - | 1 | - | ||
100 | - | - | 1 | - | ||
30 | 3 | - | - | - | ||
Nitrite (NIT) | Neg | - | 2 | 1 | 3 | |
Pos | 3 | 1 | 2 | - | ||
Leucocytes (LEU) (Leu/µL) | Neg | - | 1 | 1 | 3 | |
Ca70 | - | - | 2 | - | ||
Ca15 | 3 | 2 | - | - | ||
Glucose (GLU) (mg/dL) | Neg | 3 | 3 | 3 | 3 | |
Microscopic Examination | Epi Cells | 0 | 2 | 3 | 2 | 2 |
0-1 | 1 | - | 1 | 1 | ||
Casts | Absent | 3 | 3 | 3 | 3 | |
Crystals | Present | 3 | 3 | 3 | 3 | |
Table 9. Summary of absolute organ weights (g) record
Group, Sex & Dose (mg/kg body weight/day) |
| Adrenals | Thymus | Spleen | Testes | Epididymides | Heart | Kidneys | Brain | Liver | Prostate+Seminal vesicles with coagulating glands (PSC) |
G1, M & 0 | Mean | 0.0569 | 0.3161 | 1.1578 | 2.9419 | 0.8424 | 0.8946 | 1.7430 | 1.6811 | 7.8730 | 1.4337 |
±SD | 0.0141 | 0.0269 | 0.2774 | 0.0317 | 0.1045 | 0.1037 | 0.2386 | 0.0548 | 0.9084 | 0.4801 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, M & 100 | Mean | 0.0434 | 0.3843 | 1.3946 | 2.8097 | 0.7433 | 0.9793 | 1.7929 | 1.8006 | 8.6424 | 1.4464 |
±SD | 0.0063 | 0.0561 | 0.4100 | 0.1846 | 0.0476 | 0.0701 | 0.0379 | 0.0975 | 1.0011 | 0.1312 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, M & 300 | Mean | 0.0471 | 0.2860 | 1.4546 | 3.2638 | 0.8225 | 0.9156 | 1.7204 | 1.8688 | 8.2005 | 1.4351 |
±SD | 0.0137 | 0.0853 | 0.6647 | 0.1690 | 0.0939 | 0.1069 | 0.1870 | 0.1451 | 1.4470 | 0.1714 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, M & 1000 | Mean | 0.0502 | 0.3586 | 1.0628 | 3.2475 | 0.8604 | 0.9275 | 1.7705 | 1.8239 | 7.9500 | 1.5484 |
±SD | 0.0118 | 0.0728 | 0.2475 | 0.3785 | 0.1349 | 0.0234 | 0.0264 | 0.0664 | 0.5808 | 0.1829 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
Group, Sex & Dose (mg/kg body weight/day) |
| Adrenals | Thymus | Spleen | Ovaries with Oviduct | Uterus with cervix | Heart | Kidneys | Brain | Liver |
G1, F & 0 | Mean | 0.0653 | 0.3518 | 1.3109 | 0.1014 | 0.5245 | 0.7561 | 1.4506 | 1.6489 | 6.9631 |
±SD | 0.0155 | 0.0988 | 0.5494 | 0.0221 | 0.3749 | 0.0214 | 0.1304 | 0.0324 | 0.6644 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, F & 100 | Mean | 0.0633 | 0.2851 | 1.0969 | 0.1151 | 0.4561 | 0.8306 | 1.4629 | 1.7280 | 6.8227 |
±SD | 0.0083 | 0.0685 | 0.1513 | 0.0116 | 0.0940 | 0.1027 | 0.0496 | 0.0323 | 0.4436 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, F & 300 | Mean | 0.0643 | 0.3018 | 1.0642 | 0.1184 | 0.4597 | 0.8005 | 1.4591 | 1.7184 | 6.7091 |
±SD | 0.0019 | 0.0199 | 0.3251 | 0.0298 | 0.1173 | 0.0900 | 0.0930 | 0.0377 | 0.4766 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, F & 1000 | Mean | 0.0762 | 0.3893 | 0.9916 | 0.1098 | 0.4997 | 0.8543 | 1.4356 | 1.7151 | 7.4837 |
±SD | 0.0182 | 0.1208 | 0.1645 | 0.0163 | 0.2168 | 0.0489 | 0.1095 | 0.0883 | 0.1878 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
Table 10. Summary of organ weight relative to fasting body weight (%) record
Group, Sex & Dose (mg/kg body weight/day) | Fasting Body Weight (g) | Adrenals | Thymus | Spleen | Testes | Epididymides | Heart | Kidneys | Brain | Liver | Prostate+Seminal vesicles with coagulating glands (PSC) | |
G1, M & 0 | Mean | 236.92 | 0.0241 | 0.1343 | 0.4901 | 1.2518 | 0.3581 | 0.3783 | 0.7343 | 0.7169 | 3.3246 | 0.5969 |
±SD | 26.27 | 0.0058 | 0.0161 | 0.1172 | 0.1354 | 0.0555 | 0.0327 | 0.0185 | 0.0990 | 0.1533 | 0.1560 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, M & 100 | Mean | 237.31 | 0.0183 | 0.1614 | 0.5973 | 1.1845 | 0.3137 | 0.4127 | 0.7574 | 0.7599 | 3.6472 | 0.6141 |
±SD | 16.88 | 0.0019 | 0.0130 | 0.2070 | 0.0292 | 0.0193 | 0.0073 | 0.0408 | 0.0385 | 0.4241 | 0.0962 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, M & 300 | Mean | 235.95 | 0.0199 | 0.1200 | 0.6028 | 1.3875 | 0.3533 | 0.3876 | 0.7303 | 0.7931 | 3.4581 | 0.6079 |
±SD | 22.99 | 0.0056 | 0.0249 | 0.2143 | 0.0692 | 0.0726 | 0.0121 | 0.0612 | 0.0171 | 0.2692 | 0.0328 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, M & 1000 | Mean | 238.03 | 0.0210 | 0.1501 | 0.4493 | 1.3677 | 0.3617 | 0.3902 | 0.7446 | 0.7676 | 3.3412 | 0.6493 |
±SD | 8.58 | 0.0045 | 0.0249 | 0.1199 | 0.1890 | 0.0577 | 0.0225 | 0.0342 | 0.0545 | 0.2420 | 0.0561 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
Group, Sex & Dose (mg/kg/day) |
| Fasting Body Weight (g) | Adrenals | Thymus | Spleen | Ovaries with Oviduct | Uterus with Cervix | Heart | Kidneys | Brain | Liver |
G1, F & 0 | Mean | 181.39 | 0.0361 | 0.1930 | 0.7193 | 0.0557 | 0.2860 | 0.4168 | 0.7996 | 0.9098 | 3.8379 |
±SD | 5.09 | 0.0088 | 0.0496 | 0.2853 | 0.0110 | 0.1975 | 0.0013 | 0.0665 | 0.0434 | 0.3318 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G2, F & 100 | Mean | 192.11 | 0.0330 | 0.1498 | 0.5757 | 0.0598 | 0.2360 | 0.4313 | 0.7644 | 0.9024 | 3.5547 |
±SD | 11.58 | 0.0044 | 0.0423 | 0.1166 | 0.0025 | 0.0358 | 0.0288 | 0.0734 | 0.0730 | 0.2006 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G3, F & 300 | Mean | 182.68 | 0.0352 | 0.1651 | 0.5841 | 0.0647 | 0.2526 | 0.4388 | 0.7994 | 0.9410 | 3.6751 |
±SD | 3.97 | 0.0009 | 0.0076 | 0.1854 | 0.0152 | 0.0701 | 0.0553 | 0.0625 | 0.0302 | 0.2961 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | |
G4, F & 1000 | Mean | 192.63 | 0.0393 | 0.1997 | 0.5158 | 0.0568 | 0.2547 | 0.4455 | 0.7456 | 0.8953 | 3.9044 |
±SD | 15.88 | 0.0070 | 0.0447 | 0.0882 | 0.0039 | 0.0876 | 0.0458 | 0.0109 | 0.0995 | 0.3622 | |
n | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
Effect on fertility: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 1 000 mg/kg bw/day
- Study duration:
- subacute
- Species:
- rat
- Quality of whole database:
- One study available on the substance with a Klimisch score = 1 and other study on an analogue substance with a Klimisch score = 2.
Effect on fertility: via inhalation route
- Endpoint conclusion:
- no study available
Effect on fertility: via dermal route
- Endpoint conclusion:
- no study available
Effects on developmental toxicity
Description of key information
Read-across, test method according to OECD 422 (GLP study). Based on read-across approach from experimental data on analogue methyl acetoacetate, the NOEL for repeat dose toxicity, reproduction performance of parents and development of offspring of the test item AA-TMP was estimated to be 1109.2 mg/kg/day.
Effect on developmental toxicity: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 1 109.2 mg/kg bw/day
- Study duration:
- subacute
- Species:
- rat
- Quality of whole database:
- One study available on an analogue substance with a Klimisch score = 2.
Effect on developmental toxicity: via inhalation route
- Endpoint conclusion:
- no study available
Effect on developmental toxicity: via dermal route
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
Based on the available information, the substance is not classified for toxicity to reproduction in accordance with CLP Regulation (EC) no 1272/2008.
Additional information
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.
