Dialuminium zinc tetraoxide

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 04, 2016 - April 24, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Version / remarks:

October 3, 2008

Deviations:

yes

Remarks:

No analytic was performed as the test substance is a poor soluble inorganic substance, that is stable under the conditions. The test material was freshly prepared every day.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Dialuminium zinc tetraoxide

EC Number:

235-101-0

EC Name:

Dialuminium zinc tetraoxide

Cas Number:

12068-53-0

Molecular formula:

ZnAl2xO(3x+1), x = 1 – 9

IUPAC Name:

dialuminum; zinc(2+); oxygen(2-)

Test material form:

solid

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

SPF

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Beijing Vital River Laboratory Animal Co., LTD.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 6 - 7 weeks
- Weight at study initiation: 184.5 to 213.9 g of males and 157.4 to 186.7 g of females
- Fasting period before study: none
- Housing: ≤ 5 animals of same sex were housed in polycarbonate cages (L x W x H: 400mm x 250mm x 200 mm) bedding with corn cob.
- Diet: Irradiated rodent diet (Shanghai Puluteng Bio-technology Co., Ltd, Feed Production Certificate No: Hu Si Zheng (2014) 04001), Lot. No.: M02-F-20160419004 (valid till 18. Oct 2016); ad libitum
- Water: drinking water purified by reverse osmosis, ad libitum
- Acclimation period: 6 days

DETAILS OF FOOD AND WATER QUALITY:
The bedding material evaluation for contaminants was performed according to the national standard GB 14924.2-2001. The food evaluation for nutrients was performed by the supplier according to the Chinese national standard GB 14924.3-2010, for contaminants according to GB 14924.2-2001. The total bacteria numbers evaluation of water was performed according to the national standard for drinking water GB 5749-2006 monthly in the facility and the evaluation for contaminations was performed twice a year by the supplier according to GB 5749-2006. There were no findings that could interfere with the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.85 to 24.90, automatically controlled by air conditioner (Honeywell EBI)
- Humidity (%): 46.45 to 71.20
- Photoperiod (hrs dark / hrs light): 12/12 (from 7:30 to 19:30), artificial lighting

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Water for injection

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS
The test article was accurately weighed for each dose level, and the vehicle (control article) was added to achieve the prescribed concentrations. The suspsions were stirred with a stirrer. The formulation was stored at room temperature ( 15 - 25 °C) for the use on the day.

VEHICLE
- Amount of vehicle (if gavage): 10 mL/kg bw
- Lot/batch no. (if required): M16040405 and M16061014

Analytical verification of doses or concentrations:

no

Remarks:

No analytic was performed. The test material was freshly prepared every day. During the dosing process, the formulations were continuously stirred on the magnet mixer.

Duration of treatment / exposure:

28 days

Frequency of treatment:

Once daily, 7 days/week

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

10 male/10 female for control and high dose group (1000 mg/kg bw/day)
5 male/5 female for mid and low dose group (100 mg/kg bw/day and 300 mg/kg bw/day)

Control animals:

yes, concurrent vehicle

Details on study design:

- Rationale for animal assignment: Based on the result of a single dose exposure study (LD50 > 2000 mg/kg bw)
- Post-exposure recovery period in satellite groups: yes

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations included: mortality, consumption of food and water

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule of regular clinical observation: All animals were observed each day after dosing for signs of appearance, behavioural changes, mental state, glandular secretion, respiratory status, faecal traits and colour, genitals, mortality and other toxic manifestations.
- Time schedule of detailed clinical observations: Once before dosing, once a week from week 1 to week 4, and once in the last recovery week all animals were observed in home-cage observation, hand-held observation, open-field observation, and stimulation reflex test. The open-field observation of each animal was about 3 minutes.
- Grip strength: Once in the 4th week of treatment and last week of recovery using Grip Strength Meter.


BODY WEIGHT: Yes
- Time schedule for examinations: All animals were weighed once a week. The last measurement was performed prior to necropsy.

FOOD CONSUMPTION: Yes
- Time schedule: The quantity of food consumed in each cage was measured at weekly intervals throughout the study.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on the scheduled day of necropsy
- Anaesthetic used for blood collection: Yes, 60 mg/kg bw pentobarbital sodium, ip
- Animals fasted: Yes, 12 hours prior to blood collection
- How many animals: 5 rats/sex of each treatment group; 5 rats/sex of control group and 1000 mg/kg bw of the recovery groups
- Parameters examined: Red blood cell count (RBC), Hematocrit (HCT), Hemoglobin concentration (HGB), Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC), Reticulocyte count (RET), Reticulocyte count percentage (%RET), White blood cell count (WBC), Differential count of WBC (Neutrophil, Eosinophil, Basophil, Monocyte and Lymphocyte), Platelet count (PLT), Prothrombin time (PT), Activated partial thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on the scheduled day of necropsy
- Animals fasted: Yes, 12 hours prior to blood collection
- How many animals: 5 rats/sex of each dose groups and 5 rats/sex of control group and 1000 mg/kg bw/day group
- Parameters checked: Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Alkaline phosphatase (ALP), Creatine kinase (CK), Lactate dehydrogenase (LDH), Total bilirubin (T-Bil), Urea nitrogen (UN) ,Creatinine (Crea), Total protein (TP), Albumin/globulin ratio (A/G ratio), Albumin (ALB), Glucose (GLU), Cholesterol (CHOL), Triglycerides (TG), Sodium (Na+), Calcium (Ca), Potassium (K+), Chloride (Cl-) and Phosphorus (P)

URINALYSIS: Yes
- Time schedule for collection of urine: the last week of dosing and recovery period
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No
- Parameters checked: Urine volume, Appearance, Glucose (GLU), Bilirubin (BIL), Ketones (KET), Specific gravity (SG), Occult blood (BLO), pH, Total protein (PRO), Urobilinogen (URO), Nitrites (NIT), White blood cell (WBC)

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

OTHERS: Sexual cycle
- Time schedule for sampling: on the scheduled day of necropsy
- How many animals: all females
- Parameters checked: Vaginal Smears

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
The macroscopic appearance of the tissues of all rats was recorded and samples of the following tissues were preserved for histopathology: adrenals, aorta, brain (medullary, cerebellar and cerebral sections), breast, caecum, colon, duodenum, epididymides, eye, fallopian tube , femur (with joint, bone marrow ), heart, hypophysis, ileum (incl. Peyer's patches), jejunum, kidneys, liver, lymph nodes (mandibular and mesentric), lungs (all lobes, mainstem bronchi), lymph nodes, mesentery lymph node, oesophagus, optic nerve, ovaries, parathyroid gland, pancreas, prostate, rectum, salivary gland, sciatic nerve, seminal vesicles, skeletal muscle (biceps femoris), skin (groin), spinal cord (neck, chest, waist), spleen, stomach, testes, thymus, thyroid, trachea, urinary bladder, uterus (with cervix), vagina, other macroscopically abnormal tissue.

Testes/epididymides, eye/optic nerve were fixed in modified Davidson‘s solution. All other tissues were preserved in buffered 10% formalin.

The following ogans from each animal were weighed: brain, spleen, thymus, liver, kidneys, heart, adrenals, testis, epididymides, prostate with seminal vesicle and coagulating gland, uterus, ovary thyroid parathyroid gland


HISTOPATHOLOGY: Yes
Microsccopic examination of prepared slides (from tissues indicated under gross pathology) was carried out for all animals of the control and high-dose group.

Statistics:

Quantitative data such as body weight, food consumption (excluding recovery period data), haematology, clinical chemistry, urine volume, organ weight and ratios, number of rearing, number of faecal particles, number of urination, grip strength were presented as group mean ± standard deviation. Qualitative data (binomial categories data, unordered multi-categories data and ordinal data) such as urinary analysis, home-cage observation, hand-held observation, open-field observation (excluding number of rearing, number of faecal particles, number of urination) and stimulus response observation were presented as observed counts (frequency). When sample size was less than three, raw data of that group were directly presented without statistical comparison.
All pairwise comparison was performed between control group and each treatment group.
Analyses were performed according to sex at two-tailed probability level (α=0.05). PRISTIMA 6.1.1 was used for statistical analysis of body weight, food consumption, haematology, clinical chemistry, organ weight and ratios. The other data were statistically analyzed with Statistical Product and Service Solutions (SPSS 13.0 for Windows).
The results of histopathology were described.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

No abnormalities were observed in spontaneous activity, general state, skin and hair of all groups of animals in the dosing and recovery period.

Mortality:

no mortality observed

Description (incidence):

No unscheduled deaths occurred during the study.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No statistically significant differences were observed in all test substance groups of animals when compared to the control during the dosing and recovery period (p>0.05).

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

The food consumption was statistically significant increased for males (100 mg/kg bw/day group) on Day 7 and for females (1000 mg/kg bw/day) on recovery Day 13. A statistically significant decreased food consumption for males (1000 mg/kg bw/day) on Day 27 (p<0.05) was observed. These changes are considered to be normal test fluctuations and have no toxicological relevance.

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 statistically significant differences were observed in all groups of animals when compared to the control group during the dosing and recovery period (p>0.05).

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

Statistically significant increased glucose (GLU) levels at 300 mg/kg bw/day and 1000 mg/kg bw/day, as well as statistically significant increased phospor (P) levels at 1000 mg/kg bw/day were observed in females at the end of dosing the period when compared to the respective controls. Statistically significant decreased triglyceride (TG) levels were observed in males at 300 mg/kg bw/day at the end of dosing period compared to the control group. A statistically significant increased total bilirubin (T-Bil) level at 1000 mg/kg bw at the end of recovery was observed in males when compared to the control group. However, all these observations are not considered toxicologically relevant for the following reasons: The decrease of TG was not dose-related and the difference was slight. The increase of T-Bil occurred in the recovery period and was within the fluctuation range of animals at the same age in the laboratory (0.37 – 1.8 µmol/L). The increase of GLU and P was slight compared to the control and within the fluctuation range of females at the same age in the laboratory (GLU: 4.59 – 8.32 mmol/L, P: 1.70 – 2.96 mmol//L).

In addition, at the end of the treatment period there was no statistically significant differences between treatment and control groups in ALP, ALT, AST, urea, Crea, CK, LDH, ALB, A/G, CHOL, TP, Ca, Cl-, K+, Na+ in females and males (p> 0.05). At the end of the recovery period, the difference of blood biochemical indices at 1000 mg/kg bw/day group were not statistically significant different compared to the control groups both in females and males (p>0.05).

Urinalysis findings:

no effects observed

Description (incidence and severity):

No statistically significant difference in any of the urinary parameters between treament and control groups was observed at the end of treatment and recovery period (p>0.05).

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

Statistically significant increase in grip strength was observed in females at 1000 mg/kg bw/day in the 4th week of treatment and in males at the end of recovery period (p<0.05). Those observations are not considered toxiclogical relevant as it is an increased in grip strength only. In addition, there was no statistical significant difference from control group observed on grip strength in males at 100, 300 and 1000 mg/kg bw/day group in the 4th week of treatment as well as in females at 100 and 300 mg/kg bw/day. At the end of the recovery period treated female animals did not show a statistically significant difference compared to the controls in grip strength. Since the differences were slight and observed on grasping only, this is considered not treamtent-related.

No statistically significant differences were observed in the hand-held observations, open-field observation in animals of treatment groups compared to the control groups at the end of dosing and recovery period (p>0.05).

No statistically significant difference in any treatment group in stimulus response, i.e. alertness response, abdominal tension, auricle reflex, pinch reflex, reflexive reflex and pupil reflex was observed compared to the control group (p>0.05) on Day 5, 12 19 and 26 and on Day 12 of the recovery period.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

Statistically significant differences were observed in females at 1000 mg/kg bw/day with decreased thymus weight, thymus/body weight ratio and thymus/brain weight ratio compared to the control group at the end of the recovery period (p<0.05). The differences were slight (<= 16.85%) and it appeared in recovery period only. Furthermore, as no statistically significance was observed in the dosing period and no organ abnormalities were observed in the histopathological examinations, the observation was considered to be normal test fluctuations and are considered not toxicologically relevant.
No statistically significant differences of treated animals compared to the control groups were observed in the weight and weight ratio of other organs at the end of the treament period as well as at the end of the recovery period (p>0.05).

Slight increase of uterine weight was observed in individual females. The vaginal smear cycle examination, which was made on the day of necropsy, revealed that the animals with increased weight of the uterus were in the emotional period. Thus, this is the normal phenomenon of the estrus, not treatment related.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No treatment related gross findings were observed in brain, heart, liver, lung, kidney, spleen, gastrointestinal tract and germinal system at the end of dosing and recovery period.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No treatment related pathological alterations were observed in brain, heart, liver, lung, kidney, spleen, thymus gland, ovary, uterus, testes, epididymides, pancreas, bone marrow, gastrointestinal tract and other organs at the end of dosing and recovery period.
There were no statistically significant abnormalities observed in the treatment groups compared to control groups in the histopathological examinations in ovaries/uterus/vagina with the vaginal smear cycle examination made on the day of necropsy.

Histopathological findings: neoplastic:

no effects observed

Other effects:

not examined

Effect levels

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 adverse effects observed

Target system / organ toxicity

Key result

Critical effects observed:

no

Any other information on results incl. tables

Table 1 Effects on Body Weight of Males (g)

Detection time	Control		100 mg/kg bw/day group		300 mg/kg bw/day group		1000 mg/kg bw/day group
	n	Mean± SD	n	Mean± SD	n	Mean± SD	n	Mean± SD
Pre-dosing	10	201.5 ± 8.7	5	203.9 ± 6.5	5	203.2 ± 8.4	10	201.1 ± 8.2
Day 5	10	249.5 ± 13.4	5	249.0 ± 9.4	5	249.1 ± 10.5	10	247.4 ± 9.6
Day 12	10	303.8 ± 20.1	5	307.7 ± 15.4	5	303.4 ± 15.0	10	305.1 ± 16.5
Day 19	10	348.8 ± 25.7	5	358.8 ± 22.2	5	340.3 ± 21.5	10	350.5 ± 24.3
Day 26	10	392.4 ± 33.6	5	401.7 ± 29.0	5	383.3 ± 18.4	10	395.6 ± 33.3
Recovery day 5	5	405.7 ± 46.4	/	/	/	/	5	425.9 ± 47.3
Recovery day 12	5	429.7 ± 53.7	/	/	/	/	5	450.2 ± 51.4

Notes: * Statistical significance was noted when compared with the control (p≤0.05).

 

Table 2 Effects for D-Prism via Oral Gavage on Body Weight of Females (g)

Detection time	Control		100 mg/kg bw/day group		300 mg/kg bw/day group		1000 mg/kg bw/day group
	n	Mean± SD	n	Mean± SD	n	Mean± SD	n	Mean± SD
Pre-dosing	10	173.1 ± 7.4	5	172.6 ± 8.9	5	172.3 ± 8.9	10	174.3 ± 6.3
Day 5	10	191.7 ± 7.9	5	191.2 ± 6.3	5	184.3 ± 9.4	10	191.9 ± 4.8
Day 12	10	210.4 ± 15.0	5	213.2 ± 9.5	5	206.1 ± 9.8	10	209.8 ± 11.0
Day 19	10	225.6 ± 18.7	5	228.7 ± 11.9	5	220.8 ± 13.2	10	221.8 ± 12.3
Day 26	10	231.4 ± 20.9	5	243.6 ± 18.3	5	235.9 ± 16.6	10	232.8 ± 18.6
Recovery day 5	5	235.6 ± 27.3	/	/	/	/	5	237.3 ± 18.8
Recovery day 12	5	244.0 ± 27.4	/	/	/	/	5	245.6 ± 13.4

 

Table 3 Effects on Hematology of Males

Parameters Detection time	Control		100 mg/kg bw/day group		300 mg/kg bw/day group		1000 mg/kg bw/day group
	n	Mean± SD	n	Mean± SD	n	Mean± SD	n	Mean± SD
WBC (10^9/L)
At necropsy of the dosing period	5	5.36 ± 0.89	5	6.49 ± 1.27	5	6.05 ± 1.94	5	6.34 ± 2.59
At necropsy of the recovery period	5	5.36 ± 1.20	/	/	/	/	5	5.50 ± 1.56
NEU% (%)
At necropsy of the dosing period	5	14.8 ± 3.6	5	12.4 ± 3.5	5	14.2 ± 4.2	5	14.8 ± 2.7
At necropsy of the recovery period	5	14.3 ± 3.3	/	/	/	/	5	18.1 ± 4.7
LYM% (%)
At necropsy of the dosing period	5	81.5 ± 3.6	5	83.7 ± 4.3	5	81.8 ± 3.8	5	81.7 ± 2.7
At necropsy of the recovery period	5	81.9 ± 3.4	/	/	/	/	5	78.1 ± 5.2
MONO% (%)
At necropsy of the dosing period	5	1.6 ± 0.6	5	1.7 ± 1.0	5	1.4 ± 0.1	5	1.3 ± 0.3
At necropsy of the recovery period	5	1.8 ± 0.3	/	/	/	/	5	1.7 ± 0.6
EOS% (%)
At necropsy of the dosing period	5	0.82 ± 0.08	5	0.94 ± 0.21	5	1.04 ± 0.09	5	1.02 ± 0.47
At necropsy of the recovery period	5	0.96 ± 0.23	/	/	/	/	5	1.06 ± 0.24
BASO% (%)
At necropsy of the dosing period	5	0.22 ± 0.08	5	0.32 ± 0.11	5	0.28 ± 0.08	5	0.28 ± 0.08
At necropsy of the recovery period	5	0.14 ± 0.05	/	/	/	/	5	0.20 ± 0.07
NEU (10^9/L)
At necropsy of the dosing period	5	0.80 ± 0.27	5	0.84 ± 0.38	5	0.87 ± 0.38	5	0.88 ± 0.13
At necropsy of the recovery period	5	0.75 ± 0.18	/	/	/	/	5	1.01 ± 0.40
LYM (10^9/L)
At necropsy of the dosing period	5	4.36 ± 0.72	5	5.39 ± 0.77	5	4.94 ± 1.60	5	5.24 ± 2.38
At necropsy of the recovery period	5	4.41 ± 1.09	/	/	/	/	5	4.28 ± 1.18
MONO (10^9/L)
At necropsy of the dosing period	5	0.09 ± 0.03	5	0.12 ± 0.10	5	0.08 ± 0.03	5	0.08 ± 0.02
At necropsy of the recovery period	5	0.10 ± 0.02	/	/	/	/	5	0.10 ± 0.05
EOS (10^9/L)
At necropsy of the dosing period	5	0.046 ± 0.011	5	0.064 ± 0.022	5	0.062 ± 0.016	5	0.062 ± 0.026
At necropsy of the recovery period	5	0.052 ± 0.016	/	/	/	/	5	0.060 ± 0.016
BASO (10^9/L)
At necropsy of the dosing period	5	0.012 ± 0.004	5	0.024 ± 0.009	5	0.016 ± 0.009	5	0.020 ± 0.017
At necropsy of the recovery period	5	0.010 ± 0.000	/	/	/	/	5	0.010 ± 0.007
RBC (10^12/L)
At necropsy of the dosing period	5	7.43 ± 0.55	5	7.20 ± 0.35	5	7.36 ± 0.25	5	7.13 ± 0.40
At necropsy of the recovery period	5	7.25 ± 0.35	/	/	/	/	5	7.33 ± 0.28
HGB (g/L)
At necropsy of the dosing period	5	144 ± 8	5	145 ± 5	5	146 ± 2	5	143 ± 5
At necropsy of the recovery period	5	147 ± 4	/	/	/	/	5	148 ± 4
HCT (%)
At necropsy of the dosing period	5	44.5 ± 2.4	5	44.4 ± 2.2	5	43.9 ± 1.3	5	43.3 ± 0.6
At necropsy of the recovery period	5	41.7 ± 1.4	/	/	/	/	5	42.2 ± 1.1
MCV (fL)
At necropsy of the dosing period	5	60.1 ± 3.2	5	61.6 ± 2.0	5	59.7 ± 2.2	5	60.8 ± 2.6
At necropsy of the recovery period	5	57.6 ± 1.8	/	/	/	/	5	57.5 ± 1.2
MCH (pg)
At necropsy of the dosing period	5	19.5 ± 1.2	5	20.2 ± 0.8	5	19.9 ± 0.8	5	20.1 ± 1.6
At necropsy of the recovery period	5	20.3 ± 0.8	/	/	/	/	5	20.3 ± 0.7
MCHC (g/L)
At necropsy of the dosing period	5	324 ± 3	5	327 ± 5	5	333 ± 11	5	331 ± 14
At necropsy of the recovery period	5	352 ± 4	/	/	/	/	5	353 ± 6
PLT (10^9/L)
At necropsy of the dosing period	5	1080 ± 72	5	1087 ± 120	5	1082 ± 64	5	1084 ± 110
At necropsy of the recovery period	5	1018 ± 103	/	/	/	/	5	912 ± 73
RET % (%)
At necropsy of the dosing period	5	2.06 ± 0.40	5	2.04 ± 0.45	5	1.90 ± 0.26	5	2.30 ± 0.44
At necropsy of the recovery period	5	1.91 ± 0.45	/	/	/	/	5	1.83 ± 0.25
RET (10^12/L)
At necropsy of the dosing period	5	0.152 ± 0.027	5	0.146 ± 0.029	5	0.139 ± 0.017	5	0.166 ± 0.024
At necropsy of the recovery period	5	0.137 ± 0.024	/	/	/	/	5	0.135 ± 0.018
APTT (sec)
At necropsy of the dosing period	5	13.9 ± 1.1	5	13.1 ± 2.0	5	13.9 ± 1.9	5	13.9 ± 1.9
At necropsy of the recovery period	5	15.1 ± 2.1	/	/	/	/	5	13.9 ± 1.3
PT (sec)
At necropsy of the dosing period	5	8.4 ± 0.4	5	8.6 ± 0.4	5	8.7 ± 0.9	5	8.3 ± 0.2
At necropsy of the recovery period	5	8.9 ± 0.2	/	/	/	/	5	9.0 ± 0.4

Notes: * Statistical significance was noted when compared with the control (p≤0.05).

 

Table 4 Effects on Hematology of Females

Parameters Detection time	Control		100 mg/kg bw/daygroup		300 mg/kg bw/day group		1000 mg/kg bw/day group
	n	Mean± SD	n	Mean± SD	n	Mean± SD	n	Mean± SD
WBC (10^9/L)
At necropsy of the dosing period	5	3.84 ± 0.64	5	3.62 ± 1.06	5	3.26 ± 1.61	5	3.57 ± 0.70
At necropsy of the recovery period	5	3.81 ± 0.57	/	/	/	/	5	4.29 ± 1.50
NEU% (%)
At necropsy of the dosing period	5	13.0 ± 1.8	5	17.7 ± 4.7	5	15.9 ± 4.9	5	16.9 ± 5.4
At necropsy of the recovery period	5	14.8 ± 5.4	/	/	/	/	5	14.5 ± 6.2
LYM% (%)
At necropsy of the dosing period	5	83.6 ± 2.3	5	78.2 ± 4.6	5	80.6 ± 5.4	5	79.5 ± 6.0
At necropsy of the recovery period	5	80.4 ± 6.7	/	/	/	/	5	82.0 ± 6.0
MONO% (%)
At necropsy of the dosing period	5	1.3 ± 0.3	5	1.8 ± 0.5	5	1.4 ± 0.3	5	1.4 ± 0.6
At necropsy of the recovery period	5	1.6 ± 0.8	/	/	/	/	5	1.4 ± 0.5
EOS% (%)
At necropsy of the dosing period	5	1.12 ± 0.52	5	1.30 ± 0.10	5	1.24 ± 0.19	5	1.24 ± 0.67
At necropsy of the recovery period	5	2.02 ± 1.11	/	/	/	/	5	1.18 ± 0.45
BASO% (%)
At necropsy of the dosing period	5	0.24 ± 0.11	5	0.24 ± 0.11	5	0.18 ± 0.11	5	0.16 ± 0.05
At necropsy of the recovery period	5	0.20 ± 0.07	/	/	/	/	5	0.14 ± 0.11
NEU (10^9/L)
At necropsy of the dosing period	5	0.49 ± 0.06	5	0.61 ± 0.05	5	0.57 ± 0.47	5	0.59 ± 0.19
At necropsy of the recovery period	5	0.55 ± 0.15	/	/	/	/	5	0.60 ± 0.26
LYM (10^9/L)
At necropsy of the dosing period	5	3.22 ± 0.60	5	2.87 ± 1.00	5	2.57 ± 1.08	5	2.87 ± 0.72
At necropsy of the recovery period	5	3.09 ± 0.69	/	/	/	/	5	3.54 ± 1.35
MONO (10^9/L)
At necropsy of the dosing period	5	0.05 ± 0.01	5	0.07 ± 0.03	5	0.05 ± 0.03	5	0.05 ± 0.01
At necropsy of the recovery period	5	0.06 ± 0.02	/	/	/	/	5	0.05 ± 0.01
EOS (10^9/L)
At necropsy of the dosing period	5	0.042 ± 0.019	5	0.048 ± 0.013	5	0.044 ± 0.028	5	0.040 ± 0.012
At necropsy of the recovery period	5	0.074 ± 0.030	/	/	/	/	5	0.048 ± 0.011
BASO (10^9/L)
At necropsy of the dosing period	5	0.010 ± 0.007	5	0.010 ± 0.007	5	0.004 ± 0.005	5	0.004 ± 0.005
At necropsy of the recovery period	5	0.008 ± 0.004	/	/	/	/	5	0.006 ± 0.009
RBC (10^12/L)
At necropsy of the dosing period	5	7.03 ± 0.45	5	6.84 ± 0.22	5	6.87 ± 0.12	5	6.76 ± 0.16
At necropsy of the recovery period	5	7.13 ± 0.27	/	/	/	/	5	6.99 ± 0.18
HGB (g/L)
At necropsy of the dosing period	5	137 ± 6	5	134 ± 5	5	137 ± 6	5	134 ± 4
At necropsy of the recovery period	5	141 ± 3	/	/	/	/	5	138 ± 4
HCT (%)
At necropsy of the dosing period	5	41.2 ± 1.7	5	40.6 ± 1.4	5	40.8 ± 1.9	5	39.8 ± 1.2
At necropsy of the recovery period	5	40.2 ± 0.8	/	/	/	/	5	39.1 ± 1.0
MCV (fL)
At necropsy of the dosing period	5	58.6 ± 1.2	5	59.4 ± 1.7	5	59.4 ± 2.0	5	58.9 ± 0.9
At necropsy of the recovery period	5	56.4 ± 2.0	/	/	/	/	5	56.0 ± 0.9
MCH (pg)
At necropsy of the dosing period	5	19.6 ± 0.6	5	19.7 ± 0.5	5	19.9 ± 0.6	5	19.9 ± 0.3
At necropsy of the recovery period	5	19.8 ± 0.8	/	/	/	/	5	19.7 ± 0.6
MCHC (g/L)
At necropsy of the dosing period	5	334 ± 9	5	331 ± 4	5	336 ± 11	5	338 ± 6
At necropsy of the recovery period	5	350 ± 3	/	/	/	/	5	351 ± 6
PLT (10^9/L)
At necropsy of the dosing period	5	1150 ± 161	5	1111 ± 43	5	1061 ± 100	5	1126 ± 63
At necropsy of the recovery period	5	991 ± 156	/	/	/	/	5	961 ± 67
RET % (%)
At necropsy of the dosing period	5	2.03 ± 0.62	5	2.03 ± 0.56	5	2.14 ± 0.29	5	2.16 ± 0.64
At necropsy of the recovery period	5	1.77 ± 0.16	/	/	/	/	5	1.66 ± 0.33
RET (10^12/L)
At necropsy of the dosing period	5	0.142 ± 0.039	5	0.139 ± 0.037	5	0.147 ± 0.020	5	0.146 ± 0.041
At necropsy of the recovery period	5	0.127 ± 0.013	/	/	/	/	5	0.116 ± 0.024
APTT (sec)
At necropsy of the dosing period	5	13.2 ± 1.2	5	12.2 ± 1.8	5	12.2 ± 1.7	5	12.1 ± 1.1
At necropsy of the recovery period	5	13.7 ± 1.1	/	/	/	/	5	12.5 ± 1.8
PT (sec)
At necropsy of the dosing period	5	7.7 ± 0.5	5	7.7 ± 0.6	5	7.6 ± 0.3	5	7.7 ± 0.3
At necropsy of the recovery period	5	7.9 ± 0.6	/	/	/	/	5	8.0 ± 0.4

 

Table 5 Effects on Clinical Chemistry of Males

Parameters Detection time	Control		100 mg/kg bw/day group		300 mg/kg bw/day group		1000 mg/kg bw/day group
	n	Mean± SD	n	Mean± SD	n	Mean± SD	n	Mean± SD
ALB (g/L)
At necropsy of the dosing period	5	38.8 ± 1.2	5	38.2 ± 0.7	5	38.5 ± 1.9	5	38.3 ± 1.0
At necropsy of the recovery period	5	37.4 ± 0.9	/	/	/	/	5	38.3 ± 1.6
TP (g/L)
At necropsy of the dosing period	5	56.8 ± 1.5	5	55.3 ± 1.9	5	55.2 ± 1.6	5	55.6 ± 0.4
At necropsy of the recovery period	5	53.9 ± 0.3	/	/	/	/	5	55.3 ± 1.7
AST (U/L)
At necropsy of the dosing period	5	105.8 ± 15.2	5	107.4 ± 19.8	5	105.6 ± 7.8	5	102.3 ± 21.5
At necropsy of the recovery period	5	93.6 ± 5.7	/	/	/	/	5	88.1 ± 8.4
ALT (U/L)
At necropsy of the dosing period	5	32.2 ± 3.0	5	31.8 ± 6.8	5	34.6 ± 7.7	5	30.4 ± 7.4
At necropsy of the recovery period	5	30.6 ± 1.7	/	/	/	/	5	39.8 ± 9.0
TBIL (μmol/L)
At necropsy of the dosing period	5	0.8 ± 0.1	5	0.7 ± 0.4	5	1.0 ± 0.2	5	0.6 ± 0.1
At necropsy of the recovery period	5	1.0 ± 0.2	/	/	/	/	5	1.7 ± 0.4 *
CK (U/L)
At necropsy of the dosing period	5	676 ± 222	5	704 ± 260	5	652 ± 227	5	582 ± 141
At necropsy of the recovery period	5	439 ± 65	/	/	/	/	5	458 ± 225
CHOL (mmol/L)
At necropsy of the dosing period	5	1.30 ± 0.17	5	1.36 ± 0.37	5	1.08 ± 0.22	5	1.33 ± 0.35
At necropsy of the recovery period	5	1.29 ± 0.27	/	/	/	/	5	1.41 ± 0.38
Ca (mmol/L)
At necropsy of the dosing period	5	2.60 ± 0.03	5	2.59 ± 0.06	5	2.60 ± 0.08	5	2.59 ± 0.07
At necropsy of the recovery period	5	2.52 ± 0.05	/	/	/	/	5	2.57 ± 0.03
TG (mmol/L)
At necropsy of the dosing period	5	0.75 ± 0.22	5	0.72 ± 0.19	5	0.40 ± 0.09 *	5	0.49 ± 0.05
At necropsy of the recovery period	5	0.69 ± 0.19	/	/	/	/	5	0.67 ± 0.21
Crea (μmol/L)
At necropsy of the dosing period	5	18.8 ± 2.2	5	18.0 ± 2.5	5	17.4 ± 1.1	5	19.6 ± 3.3
At necropsy of the recovery period	5	19.6 ± 2.6	/	/	/	/	5	20.2 ± 0.8
GLU (mmol/L)
At necropsy of the dosing period	5	6.85 ± 0.29	5	7.14 ± 0.57	5	6.26 ± 0.38	5	7.08 ± 0.67
At necropsy of the recovery period	5	6.48 ± 0.45	/	/	/	/	5	7.31 ± 1.08
Urea (mmol/L)
At necropsy of the dosing period	5	4.72 ± 0.41	5	4.70 ± 0.66	5	4.67 ± 0.63	5	4.80 ± 1.02
At necropsy of the recovery period	5	4.32 ± 0.51	/	/	/	/	5	4.87 ± 0.64
P (mmol/L)
At necropsy of the dosing period	5	2.76 ± 0.09	5	2.72 ± 0.09	5	2.73 ± 0.20	5	2.95 ± 0.15
At necropsy of the recovery period	5	2.30 ± 0.08	/	/	/	/	5	2.46 ± 0.14
ALP (U/L)
At necropsy of the dosing period	5	150.3 ± 25.1	5	149.2 ± 22.0	5	165.3 ± 45.0	5	164.8 ± 34.3
At necropsy of the recovery period	5	123.3 ± 18.6	/	/	/	/	5	137.9 ± 35.7
LDH (U/L)
At necropsy of the dosing period	5	1137 ± 333	5	1128 ± 388	5	1087 ± 340	5	1060 ± 352
At necropsy of the recovery period	5	868 ± 160	/	/	/	/	5	654 ± 159
K^+(mmol/L)
At necropsy of the dosing period	5	4.56 ± 0.28	5	4.66 ± 0.16	5	4.51 ± 0.04	5	4.53 ± 0.12
At necropsy of the recovery period	5	4.49 ± 0.15	/	/	/	/	5	4.44 ± 0.12
Na^+(mmol/L)
At necropsy of the dosing period	5	143 ± 1	5	143 ± 0	5	144 ± 1	5	144 ± 1
At necropsy of the recovery period	5	144 ± 2	/	/	/	/	5	142 ± 1
Cl^-(mmol/L)
At necropsy of the dosing period	5	101.8 ± 1.2	5	102.8 ± 0.5	5	102.3 ± 1.1	5	101.9 ± 0.7
At necropsy of the recovery period	5	101.4 ± 1.3	/	/	/	/	5	100.6 ± 1.3
A/G
At necropsy of the dosing period	5	2.16 ± 0.10	5	2.26 ± 0.20	5	2.32 ± 0.18	5	2.22 ± 0.17
At necropsy of the recovery period	5	2.29 ± 0.19	/	/	/	/	5	2.25 ± 0.18

Notes: * Statistical significance was noted when compared with the control (p≤0.05).

 

Table 6 Effects on Clinical Chemistry of Females

Parameters Detection time	Control		100 mg/kg bw/day group		300 mg/kg bw/day group		1000 mg/kg bw/day group
	n	Mean± SD	n	Mean± SD	n	Mean± SD	n	Mean± SD
ALB (g/L)
At necropsy of the dosing period	5	43.9 ± 3.5	5	44.7 ± 1.5	5	42.9 ± 2.7	5	43.6 ± 2.2
At necropsy of the recovery period	5	46.1 ± 3.3	/	/	/	/	5	42.4 ± 4.7
TP (g/L)
At necropsy of the dosing period	5	61.3 ± 4.0	5	59.5 ± 2.4	5	58.3 ± 1.8	5	58.2 ± 3.6
At necropsy of the recovery period	5	61.9 ± 3.8	/	/	/	/	5	57.6 ± 4.0
AST (U/L)
At necropsy of the dosing period	5	105.8 ± 37.2	5	99.8 ± 16.3	5	105.5 ± 3.2	5	97.9 ± 11.8
At necropsy of the recovery period	5	93.3 ± 14.1	/	/	/	/	5	93.5 ± 18.7
ALT (U/L)
At necropsy of the dosing period	5	21.8 ± 4.0	5	25.2 ± 7.2	5	25.0 ± 5.7	5	25.8 ± 7.7
At necropsy of the recovery period	5	28.0 ± 7.0	/	/	/	/	5	29.6 ± 6.1
TBIL (μmol/L)
At necropsy of the dosing period	5	1.1 ± 0.2	5	1.1 ± 0.6	5	1.1 ± 0.5	5	1.5 ± 0.3
At necropsy of the recovery period	5	1.2 ± 0.3	/	/	/	/	5	0.9 ± 0.1
CK (U/L)
At necropsy of the dosing period	5	763 ± 537	5	555 ± 178	5	658 ± 95	5	526 ± 71
At necropsy of the recovery period	5	486 ± 174	/	/	/	/	5	435 ± 142
CHOL (mmol/L)
At necropsy of the dosing period	5	1.50 ± 0.29	5	1.53 ± 0.36	5	1.56 ± 0.59	5	1.55 ± 0.42
At necropsy of the recovery period	5	1.55 ± 0.09	/	/	/	/	5	1.40 ± 0.20
Ca (mmol/L)
At necropsy of the dosing period	5	2.49 ± 0.06	5	2.56 ± 0.06	5	2.50 ± 0.06	5	2.58 ± 0.07
At necropsy of the recovery period	5	2.62 ± 0.06	/	/	/	/	5	2.60 ± 0.03
TG (mmol/L)
At necropsy of the dosing period	5	0.34 ± 0.17	5	0.24 ± 0.07	5	0.23 ± 0.11	5	0.24 ± 0.08
At necropsy of the recovery period	5	0.31 ± 0.06	/	/	/	/	5	0.45 ± 0.30
Crea (μmol/L)
At necropsy of the dosing period	5	26.0 ± 2.3	5	26.4 ± 4.7	5	24.8 ± 2.9	5	25.0 ± 2.5
At necropsy of the recovery period	5	28.0 ± 4.2	/	/	/	/	5	31.8 ± 5.3
GLU (mmol/L)
At necropsy of the dosing period	5	5.90 ± 0.15	5	6.40 ± 0.64	5	6.85 ± 0.27 *	5	6.41 ± 0.20 *
At necropsy of the recovery period	5	6.55 ± 0.14	/	/	/	/	5	6.77 ± 0.54
Urea (mmol/L)
At necropsy of the dosing period	5	4.82 ± 0.48	5	4.64 ± 0.41	5	5.11 ± 0.97	5	5.06 ± 0.37
At necropsy of the recovery period	5	5.32 ± 0.64	/	/	/	/	5	6.38 ± 1.61
P (mmol/L)
At necropsy of the dosing period	5	2.11 ± 0.11	5	2.19 ± 0.22	5	2.17 ± 0.14	5	2.54 ± 0.13 *
At necropsy of the recovery period	5	1.88 ± 0.22	/	/	/	/	5	2.05 ± 0.34
ALP (U/L)
At necropsy of the dosing period	5	72.5 ± 17.6	5	73.7 ± 17.2	5	72.2 ± 16.5	5	84.1 ± 33.1
At necropsy of the recovery period	5	44.9 ± 11.7	/	/	/	/	5	57.4 ± 12.0
LDH (U/L)
At necropsy of the dosing period	5	1093 ± 629	5	1019 ± 328	5	1150 ± 288	5	984 ± 180
At necropsy of the recovery period	5	934 ± 323	/	/	/	/	5	878 ± 311
K^+(mmol/L)
At necropsy of the dosing period	5	4.13 ± 0.19	5	4.14 ± 0.25	5	3.95 ± 0.13	5	4.10 ± 0.22
At necropsy of the recovery period	5	3.98 ± 0.25	/	/	/	/	5	4.00 ± 0.29
Na^+(mmol/L)
At necropsy of the dosing period	5	143 ± 1	5	143 ± 1	5	143 ± 1	5	144 ± 1
At necropsy of the recovery period	5	142 ± 1	/	/	/	/	5	143 ± 1
Cl^-(mmol/L)
At necropsy of the dosing period	5	102.7 ± 1.6	5	103.3 ± 1.2	5	103.7 ± 1.2	5	103.8 ± 0.7
At necropsy of the recovery period	5	101.1 ± 0.5	/	/	/	/	5	101.8 ± 1.2
A/G
At necropsy of the dosing period	5	2.54 ± 0.28	5	3.03 ± 0.17	5	2.81 ± 0.39	5	3.04 ± 0.43
At necropsy of the recovery period	5	2.91 ± 0.27	/	/	/	/	5	2.80 ± 0.45

 

Table 7 Effects of D-Prism via Oral Gavage on Urine parameters of Males

Parameters	Detection time Grade	Control	100 mg/kg bw/day group	300 mg/kg bw/day group	1000 mg/kg bw/day group
Urine color
	Day 28
	Yellow	5	5	5	5
	Recovery day 14
	Yellow	5	/	/	4
	Dull yellow	0	/	/	1
Urine clarity
	Day 28
	Clear	4	2	4	2
	Turbidness	1	3	1	3
	Recovery day 14
	Clear	2	/	/	1
	Turbidness	3	/	/	4
GLU(mmol/L)
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	5
BIL
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	5
KET(mmol/L)
	Day 28
	-	5	4	5	4
	1.5	0	1	0	1
	Recovery day 14
	-	2	/	/	4
	1.5	3	/	/	1
SG
	Day 28
	<=1.005	0	0	2	0
	1.010	1	2	0	0
	1.015	1	2	1	5
	1.020	2	1	2	0
	1.025	1	0	0	0
	Recovery day 14
	<=1.005	1	/	/	0
	1.010	0	/	/	1
	1.015	2	/	/	1
	1.020	0	/	/	1
	1.025	1	/	/	0
	>1.030	1	/	/	2
BLO(Ery/μL)
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	5
pH
	Day 28
	7.5	0	1	1	0
	8.0	4	4	3	4
	8.5	1	0	1	1
	Recovery day 14
	6.0	1	/	/	1
	8.0	2	/	/	0
	8.5	2	/	/	4
PRO(g/L)
	Day 28
	-	5	5	4	3
	0.3	0	0	1	2
	Recovery day 14
	-	2	/	/	3
	0.3	3	/	/	2
URO (μmol/L)
	Day 28
	3.2	5	5	5	5
	Recovery day 14
	3.2	5	/	/	5
NIT
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	4
	+	0	/	/	1
WBC(Leu/μL)
	Day 28
	-	4	5	5	5
	15	1	0	0	0
	Recovery day 14
	-	5	/	/	3
	15	0	/	/	2

Notes: * Statistical significance was noted when compared with the control (P≤0.05).

 

Table 8 Effects of D-Prism via Oral Gavage on Urine parameters of Females

Parameters	Detection time Grade	Control	100 mg/kg bw/day group	300 mg/kg bw/day group	1000 mg/kg bw/day group
Urine color
	Day 28
	Yellow	5	5	5	5
	Recovery day 14
	Yellow	5	/	/	5
Urine clarity
	Day 28
	Clear	5	5	5	5
	Recovery day 14
	Clear	4	/	/	4
	Turbidness	0	/	/	1
	Dirty	1	/	/	0
GLU(mmol/L)
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	5
BIL
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	5
KET(mmol/L)
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	5
SG
	Day 28
	<=1.005	2	2	2	1
	1.010	2	3	3	0
	1.015	1	0	0	3
	1.020	0	0	0	1
	Recovery day 14
	<=1.005	1	/	/	2
	1.010	2	/	/	1
	1.015	1	/	/	1
	1.020	0	/	/	1
	1.025	1	/	/	0
BLO(Ery/μL)
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	5
pH
	Day 28
	7.5	0	0	1	1
	8.0	4	4	3	4
	8.5	1	1	1	0
	Recovery day 14
	6.5	1	/	/	1
	7.0	0	/	/	2
	7.5	1	/	/	0
	8.0	3	/	/	1
	8.5	0	/	/	1
PRO(g/L)
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	5
URO (μmol/L)
	Day 28
	3.2	5	5	5	5
	Recovery day 14
	3.2	5	/	/	5
NIT
	Day 28
	-	4	4	4	4
	+	1	1	1	1
	Recovery day 14
	-	4	/	/	4
	+	1	/	/	1
WBC(Leu/μL)
	Day 28
	-	5	5	5	5
	Recovery day 14
	-	5	/	/	4
	15	0	/	/	1

 

Table 9 Effects on Organ Weight and Ratios of Males

Parameters Detection time	Control		100 mg/kg bw/day group		300 mg/kg bw/day group		1000 mg/kg bw/day group
	n	Mean± SD	n	Mean± SD	n	Mean± SD	n	Mean± SD
Terminal Body weight (g)
End of Dosingperiod	5	385.860 ± 21.770	5	387.180 ± 31.442	5	364.880 ± 17.616	5	375.640 ± 25.621
End of recovery period	5	410.240 ± 50.549	/	/	/	/	5	436.600 ± 51.849
Organ weight (g)
Thyroid and parathyroid glands
End of Dosing period	5	0.0238 ± 0.0036	5	0.0254 ± 0.0036	5	0.0242 ± 0.0045	5	0.0193 ± 0.0049
End of recovery period	5	0.0292 ± 0.0074	/	/	/	/	5	0.0326 ± 0.0043
Liver
End of Dosing period	5	11.433 ± 1.055	5	11.611 ± 1.741	5	10.327 ± 1.220	5	10.679 ± 0.960
End of recovery period	5	11.171 ± 1.270	/	/	/	/	5	12.166 ± 1.971
Spleen
End of Dosing period	5	0.772 ± 0.084	5	0.857 ± 0.184	5	0.852 ± 0.077	5	0.804 ± 0.110
End of recovery period	5	0.715 ± 0.099	/	/	/	/	5	0.855 ± 0.115
Kidney
End of Dosing period	5	2.670 ± 0.153	5	2.765 ± 0.350	5	2.705 ± 0.165	5	2.534 ± 0.295
End of recovery period	5	2.904 ± 0.363	/	/	/	/	5	2.963 ± 0.324
Adrenal gland
End of Dosing period	5	0.057 ± 0.005	5	0.056 ± 0.004	5	0.061 ± 0.011	5	0.048 ± 0.009
End of recovery period	5	0.049 ± 0.009	/	/	/	/	5	0.053 ± 0.008
Testis
End of Dosing period	5	2.846 ± 0.843	5	2.809 ± 1.175	5	3.251 ± 0.207	5	3.112 ± 0.299
End of recovery period	5	3.368 ± 0.156	/	/	/	/	5	3.200 ± 0.278
Epididymis
End of Dosing period	5	0.981 ± 0.089	5	0.897 ± 0.196	5	0.946 ± 0.109	5	0.910 ± 0.074
End of recovery period	5	1.176 ± 0.057	/	/	/	/	5	1.183 ± 0.097
Thymus gland
End of Dosing period	5	0.605 ± 0.112	5	0.632 ± 0.117	5	0.537 ± 0.074	5	0.591 ± 0.202
End of recovery period	5	0.489 ± 0.135	/	/	/	/	5	0.456 ± 0.134
Heart
End of Dosing period	5	1.277 ± 0.110	5	1.427 ± 0.391	5	1.333 ± 0.114	5	1.247 ± 0.101
End of recovery period	5	1.377 ± 0.161	/	/	/	/	5	1.446 ± 0.184
Brain
End of Dosing period	5	2.031 ± 0.084	5	1.972 ± 0.097	5	1.975 ± 0.013	5	1.937 ± 0.102
End of recovery period	5	1.980 ± 0.122	/	/	/	/	5	2.054 ± 0.080
Prostate + seminal vesicle + coagulation gland
End of Dosing period	5	2.295 ± 0.253	5	2.102 ± 0.215	5	2.290 ± 0.351	5	2.117 ± 0.191
End of recovery period	5	2.996 ± 0.302					5	2.744 ± 0.383
Organ/Body Weight Ratio (%)
Thyroid and parathyroid glands
End of Dosing period	5	0.0061 ± 0.0006	5	0.0066 ± 0.0013	5	0.0066 ± 0.0013	5	0.0051 ± 0.0013
End of recovery period	5	0.0071 ± 0.0015	/	/	/	/	5	0.0076 ± 0.0018
Liver
End of Dosing period	5	2.962 ± 0.200	5	2.989 ± 0.284	5	2.826 ± 0.229	5	2.843 ± 0.177
End of recovery period	5	2.727 ± 0.089	/	/	/	/	5	2.781 ± 0.258
Spleen
End of Dosing period	5	0.200 ± 0.025	5	0.220 ± 0.037	5	0.234 ± 0.021	5	0.215 ± 0.032
End of recovery period	5	0.177 ± 0.032	/	/	/	/	5	0.199 ± 0.044
Kidney
End of Dosing period	5	0.692 ± 0.029	5	0.713 ± 0.042	5	0.741 ± 0.025	5	0.673 ± 0.046
End of recovery period	5	0.710 ± 0.069	/	/	/	/	5	0.680 ± 0.018
Adrenal gland
End of Dosing period	5	0.015 ± 0.002	5	0.015 ± 0.002	5	0.017 ± 0.003	5	0.013 ± 0.003
End of recovery period	5	0.012 ± 0.002	/	/	/	/	5	0.012 ± 0.002
Testis
End of Dosing period	5	0.735 ± 0.204	5	0.721 ± 0.289	5	0.891 ± 0.020	5	0.831 ± 0.085
End of recovery period	5	0.832 ± 0.118	/	/	/	/	5	0.739 ± 0.082
Epididymis
End of Dosing period	5	0.254 ± 0.016	5	0.233 ± 0.055	5	0.259 ± 0.022	5	0.243 ± 0.017
End of recovery period	5	0.290 ± 0.034	/	/	/	/	5	0.275 ± 0.044
Thymus gland
End of Dosing period	5	0.156 ± 0.023	5	0.163 ± 0.029	5	0.147 ± 0.018	5	0.156 ± 0.045
End of recovery period	5	0.118 ± 0.021	/	/	/	/	5	0.104 ± 0.023
Heart
End of Dosing period	5	0.331 ± 0.014	5	0.366 ± 0.083	5	0.365 ± 0.024	5	0.332 ± 0.023
End of recovery period	5	0.336 ± 0.020	/	/	/	/	5	0.331 ± 0.006
Brain
End of Dosing period	5	0.528 ± 0.039	5	0.511 ± 0.027	5	0.542 ± 0.026	5	0.516 ± 0.019
End of recovery period	5	0.488 ± 0.056	/	/	/	/	5	0.476 ± 0.056
Prostate + seminal vesicle + coagulation gland
End of Dosing period	5	0.596 ± 0.071	5	0.543 ± 0.026	5	0.626 ± 0.071	5	0.565 ± 0.050
End of recovery period	5	0.735 ± 0.072	/	/	/	/	5	0.633 ± 0.095
Organ/Brain Weight Ratio (%)
Thyroid and parathyroid glands
End of Dosing period	5	1.174 ± 0.206	5	1.293 ± 0.196	5	1.227 ± 0.223	5	0.999 ± 0.265
End of recovery period	5	1.469 ± 0.363	/	/	/	/	5	1.583 ± 0.191
Liver
End of Dosing period	5	563.56 ± 55.53	5	588.41 ± 81.37	5	523.15 ± 63.32	5	551.35 ± 43.01
End of recovery period	5	564.14 ± 56.55	/	/	/	/	5	591.17 ± 84.69
Spleen
End of Dosing period	5	37.98 ± 3.77	5	43.30 ± 8.26	5	43.14 ± 3.83	5	41.55 ± 5.81
End of recovery period	5	36.08 ± 4.06	/	/	/	/	5	41.59 ± 5.30
Kidney
End of Dosing period	5	131.68 ± 10.62	5	139.85 ± 11.34	5	137.03 ± 8.71	5	130.57 ± 10.20
End of recovery period	5	146.27 ± 11.58	/	/	/	/	5	144.26 ± 15.15
Adrenal gland
End of Dosing period	5	2.82 ± 0.36	5	2.84 ± 0.30	5	3.08 ± 0.54	5	2.48 ± 0.54
End of recovery period	5	2.47 ± 0.41	/	/	/	/	5	2.59 ± 0.34
Testis
End of Dosing period	5	141.05 ± 43.73	5	141.01 ± 56.59	5	164.66 ± 10.67	5	160.62 ± 12.05
End of recovery period	5	170.47 ± 10.68	/	/	/	/	5	156.18 ± 17.65
Epididymis
End of Dosing period	5	48.43 ± 5.67	5	45.37 ± 9.38	5	47.92 ± 5.64	5	46.93 ± 2.19
End of recovery period	5	59.42 ± 1.67	/	/	/	/	5	57.68 ± 5.86
Thymus gland
End of Dosing period	5	29.90 ± 6.22	5	32.10 ± 6.42	5	27.22 ± 3.84	5	30.44 ± 9.85
End of recovery period	5	24.85 ± 7.58	/	/	/	/	5	22.02 ± 5.73
Heart
End of Dosing period	5	63.00 ± 6.98	5	71.93 ± 16.98	5	67.52 ± 6.07	5	64.32 ± 2.83
End of recovery period	5	69.56 ± 7.49	/	/	/	/	5	70.30 ± 7.77
Prostate + seminal vesicle + coagulation gland
End of Dosing period	5	113.31 ± 15.53	5	106.41 ± 6.45	5	115.97 ± 17.93	5	109.34 ± 8.80
End of recovery period	5	151.15 ± 9.34	/	/	/	/	5	133.66 ± 19.02

Notes: * Statistical significance was noted when compared with the control (p≤0.05).

 

Table 10 Effects of D-Prism via Oral Gavage on Organ Weight and Ratios of Females

Parameters Detection time	Control		100 mg/kg bw/day group		300 mg/kg bw/day group		1000 mg/kg bw/day group
	n	Mean± SD	n	Mean± SD	n	Mean± SD	n	Mean± SD
Terminal Body weight (g)
End of Dosing period	5	228.660 ± 15.507	5	230.340 ± 16.748	5	223.860 ± 20.493	5	227.640 ± 18.307
End of recovery period	5	228.840 ± 26.364	/	/	/	/	5	228.900 ± 11.755
Organ weight (g)
Thyroid and parathyroid glands
End of Dosing period	5	0.0189 ± 0.0046	5	0.0192 ± 0.0029	5	0.0245 ± 0.0039	5	0.0195 ± 0.0063
End of recovery period	5	0.0299 ± 0.0120	/	/	/	/	5	0.0221 ± 0.0057
Liver
End of Dosing period	5	6.431 ± 0.699	5	6.350 ± 0.492	5	6.614 ± 0.648	5	6.044 ± 0.307
End of recovery period	5	6.192 ± 0.787	/	/	/	/	5	6.128 ± 0.568
Spleen
End of Dosing period	5	0.646 ± 0.212	5	0.572 ± 0.135	5	0.673 ± 0.194	5	0.622 ± 0.065
End of recovery period	5	0.540 ± 0.119	/	/	/	/	5	0.547 ± 0.071
Kidney
End of Dosing period	5	1.739 ± 0.095	5	1.698 ± 0.148	5	1.723 ± 0.121	5	1.666 ± 0.185
End of recovery period	5	1.633 ± 0.309	/	/	/	/	5	1.619 ± 0.052
Adrenal gland
End of Dosing period	5	0.062 ± 0.004	5	0.064 ± 0.008	5	0.062 ± 0.009	5	0.059 ± 0.009
End of recovery period	5	0.061 ± 0.005	/	/	/	/	5	0.055 ± 0.009
Ovary
End of Dosing period	5	0.102 ± 0.009	5	0.099 ± 0.012	5	0.104 ± 0.020	5	0.097 ± 0.016
End of recovery period	5	0.092 ± 0.011	/	/	/	/	5	0.085 ± 0.011
Uterus
End of Dosing period	5	0.616 ± 0.147	5	0.567 ± 0.127	5	0.545 ± 0.097	5	0.578 ± 0.239
End of recovery period	5	0.447 ± 0.071	/	/	/	/	5	0.548 ± 0.167
Thymus gland
End of Dosing period	5	0.470 ± 0.124	5	0.412 ± 0.060	5	0.445 ± 0.067	5	0.400 ± 0.119
End of recovery period	5	0.417 ± 0.029	/	/	/	/	5	0.349 ± 0.053 *
Heart
End of Dosing period	5	0.854 ± 0.068	5	0.876 ± 0.075	5	0.859 ± 0.155	5	0.840 ± 0.132
End of recovery period	5	0.865 ± 0.111	/	/	/	/	5	0.822 ± 0.077
Brain
End of Dosing period	5	1.823 ± 0.097	5	1.822 ± 0.106	5	1.855 ± 0.071	5	1.896 ± 0.055
End of recovery period	5	1.813 ± 0.062	/	/	/	/	5	1.816 ± 0.094
Organ/Body Weight Ratio (%)
Thyroid and parathyroid glands
End of Dosing period	5	0.0082 ± 0.0015	5	0.0084 ± 0.0012	5	0.0110 ± 0.0016	5	0.0086 ± 0.0027
End of recovery period	5	0.0129 ± 0.0049	/	/	/	/	5	0.0097 ± 0.0028
Liver
End of Dosing period	5	2.807 ± 0.122	5	2.758 ± 0.113	5	2.960 ± 0.230	5	2.662 ± 0.125
End of recovery period	5	2.709 ± 0.213	/	/	/	/	5	2.679 ± 0.228
Spleen
End of Dosing period	5	0.286 ± 0.102	5	0.247 ± 0.044	5	0.297 ± 0.057	5	0.275 ± 0.036
End of recovery period	5	0.236 ± 0.043	/	/	/	/	5	0.238 ± 0.021
Kidney
End of Dosing period	5	0.761 ± 0.028	5	0.737 ± 0.028	5	0.771 ± 0.032	5	0.731 ± 0.039
End of recovery period	5	0.710 ± 0.067	/	/	/	/	5	0.708 ± 0.021
Adrenal gland
End of Dosing period	5	0.027 ± 0.002	5	0.028 ± 0.005	5	0.028 ± 0.005	5	0.026 ± 0.003
End of recovery period	5	0.027 ± 0.004	/	/	/	/	5	0.024 ± 0.004
Ovary
End of Dosing period	5	0.045 ± 0.005	5	0.043 ± 0.006	5	0.047 ± 0.009	5	0.043 ± 0.008
End of recovery period	5	0.041 ± 0.004	/	/	/	/	5	0.037 ± 0.005
Uterus
End of Dosing period	5	0.271 ± 0.073	5	0.245 ± 0.050	5	0.247 ± 0.056	5	0.253 ± 0.097
End of recovery period	5	0.195 ± 0.022	/	/	/	/	5	0.238 ± 0.065
Thymus gland
End of Dosing period	5	0.208 ± 0.061	5	0.179 ± 0.025	5	0.199 ± 0.028	5	0.174 ± 0.042
End of recovery period	5	0.184 ± 0.015	/	/	/	/	5	0.153 ± 0.022 *
Heart
End of Dosing period	5	0.374 ± 0.033	5	0.381 ± 0.032	5	0.382 ± 0.043	5	0.368 ± 0.034
End of recovery period	5	0.379 ± 0.039	/	/	/	/	5	0.359 ± 0.021
Brain
End of Dosing period	5	0.799 ± 0.054	5	0.793 ± 0.060	5	0.834 ± 0.071	5	0.837 ± 0.072
End of recovery period	5	0.800 ± 0.083	/	/	/	/	5	0.795 ± 0.047
Organ/Brain Weight Ratio (%)
Thyroid and parathyroid glands
End of Dosing period	5	1.034 ± 0.211	5	1.053 ± 0.128	5	1.326 ± 0.235	5	1.036 ± 0.359
End of recovery period	5	1.638 ± 0.649	/	/	/	/	5	1.217 ± 0.304
Liver
End of Dosing period	5	352.67 ± 31.78	5	349.18 ± 28.85	5	356.34 ± 29.81	5	319.16 ± 22.59
End of recovery period	5	341.10 ± 39.02	/	/	/	/	5	337.94 ± 33.09
Spleen
End of Dosing period	5	35.25 ± 10.38	5	31.44 ± 7.26	5	36.20 ± 10.06	5	32.88 ± 4.21
End of recovery period	5	29.65 ± 5.83	/	/	/	/	5	30.16 ± 4.09
Kidney
End of Dosing period	5	95.48 ± 5.64	5	93.46 ± 9.96	5	92.93 ± 6.23	5	87.98 ± 10.84
End of recovery period	5	89.92 ± 16.23	/	/	/	/	5	89.29 ± 5.25
Adrenal gland
End of Dosing period	5	3.38 ± 0.27	5	3.52 ± 0.61	5	3.33 ± 0.56	5	3.13 ± 0.48
End of recovery period	5	3.34 ± 0.31	/	/	/	/	5	3.00 ± 0.36
Ovary
End of Dosing period	5	5.60 ± 0.59	5	5.45 ± 0.88	5	5.62 ± 1.01	5	5.15 ± 0.90
End of recovery period	5	5.10 ± 0.60	/	/	/	/	5	4.68 ± 0.79
Uterus
End of Dosing period	5	33.95 ± 8.87	5	31.15 ± 6.79	5	29.34 ± 4.92	5	30.38 ± 12.26
End of recovery period	5	24.65 ± 3.89	/	/	/	/	5	30.25 ± 9.62
Thymus gland
End of Dosing period	5	26.07 ± 8.01	5	22.62 ± 3.05	5	23.91 ± 2.97	5	21.12 ± 6.33
End of recovery period	5	23.03 ± 1.68	/	/	/	/	5	19.28 ± 3.06 *
Heart
End of Dosing period	5	46.90 ± 4.39	5	48.18 ± 4.35	5	46.25 ± 7.68	5	44.29 ± 6.54
End of recovery period	5	47.65 ± 5.65	/	/	/	/	5	45.31 ± 4.38

 

Applicant's summary and conclusion

Conclusions:

As no adverse effects were observed up to the highest dose group the NOAEL was considered to be >= 1000 mg/kg bw/day for female and male rats.