Aluminum magnesium vanadium oxide

Endpoint summary

Description of key information

Oral: Repeated dose 90-Day oral toxicity study in rodents (OECD 408) rat: NOAEL (systemic) = 1000 mg/kg bw/day (m/f)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 Nov 2014 to 23 Apr 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

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

Remarks:

National (CFDA) GLP certificate

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Experimental Animal Center of Academy of Military Medical Sciences
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 6-8 weeks
- Weight at study initiation: Male 186.1 ± 7.9 g, female 165.2 ± 5.8 g.
- Fasting period before study: Non- fasted animals at start of dosing
- Housing: The rats were housed in a SPF standard barrier facility.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Animals were quarantined for an acclimatization period of seven days.

DETAILS OF FOOD AND WATER QUALITY:
The SPF mouse/rat maintenance diet was provided by the Experimental Animal Center of Academy of Military Medical Sciences (license No: SCXK (Army) 2012-0003). The analyses report of nutrients and contaminants were provided by diet manufacturers in every quarter of year.
The results of representative reports showed that the nutrients and pollutants level was in comply with national standards (GB 14924.2-2001 Laboratory animals-Hygienic standard for formula feeds and GB 14924.3-2010 Laboratory animals-Nutrients for formula feeds) without interference or impact on the study.
Animal drinking water was filter-sterilized prepared by water filtered machine. Rats had free access to water. The microbes and other indicators of drinking water were analyzed twice a year. The results of representative water quality analytical reports showed that the microbesand pollutant contaminants level was in compliance with national standard (GB5749-2006 Standards for drinking water quality) without interference or impact on study.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25°C
- Humidity (%): 40-70%
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): day/night cycle was 12 hours (12 hours light from 06:00-18:00, 12 hours dark from 18:00-06:00)

IN-LIFE DATES: From: 8 Nov 2014 To:5 Feb 2015

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test item formulations were freshly prepared every day immediately prior to application. Appropriate amount of the test item was weighted into a tared glass vial on a suitable precision balance and the vehicle (corn oil) was added to obtain the designed final concentration of the test item formulations. Homogeneity of the test item in the vehicle was maintained by vortexing the prepared suspension thoroughly before dose administration.
The vehicle (corn oil) was used as the control formulation.


VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil is frequently used as a suitable vehicle in animal experiment when the solubility of test substance is low in water. Moreover, coin oil has been proposed as a suitable vehicle by the OECD (OECD Guidelines for Testing of Chemicals: 407 repeated dose 28-day oral toxicity study in rodents, Oct 2008). Sponsor has agreed to use corn oil as vehicle in this study.
- Concentration in vehicle: 25, 75 and 250 mg/mL
- Amount of vehicle (if gavage): 0.4 mL/100 g bw
- Lot/batch no. (if required): QS130302011011
- Purity: according to the nactional guidance on maize oil (GB 19111-2003)

Analytical verification of doses or concentrations:

no

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.

Duration of treatment / exposure:

28 days

Frequency of treatment:

Once daily, continuously

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Low dose (LD) group

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Medium dose(MD) group

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

High dose (HD) group

No. of animals per sex per dose:

5 aminals per sex per dose in control(C), LD, MD, HD,control recovery (CR) and HDR group

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: In consultation with the sponsor and on the basis of toxicology background information, LC50 (inhalation) > 5mg/L; LD50 (dermal) >2000 mg/kg body weight; LD50 (oral) > 2000mg/kg body weight; no target organ toxicity after treatment with single sublethal doses; no skin and eye irritation; no skin sensitization; no mutagenicity.

- Post-exposure recovery period in satellite groups: 14 days

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once a day during the treatment and recovery period
- Cage side observations included: mortality/viability

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly during the treatment and recovery period

BODY WEIGHT: Yes
- Time schedule for examinations: Once by animal receiving; Once before experimental grouping; Weekly during the treatmentand recovery period; Once before necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Weekly during the treatment and recovery period.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

FOOD EFFICIENCY:No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood:at the end of treatment period and recovery period
- Anaesthetic used for blood collection: Yes
- Animals fasted: Yes
- How many animals: all
- Parameters examined: red blood cell (RBC), hemoglobin (HGB), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelet (PLT), platelet distribution width (PDW), hematocrit (HCT), mean corpuscular volume (MCV), red cell distribution width (RDW), mean platelet volume (MPV), white blood cell (WBC) and classification.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of treatment period and recovery period
- Animals fasted: Yes
- How many animals: all
- Parameters examined: aspartate amino transferase (AST), alanine amino transterase (ALT), alkaline phosphatase (ALP), creatine kinase (CK), total protein (TP), albumin (ALB), total bilirubin (TB), direct bilirubin (DB), triglycerides (TG), urea (Urea), total cholesterol (TC), glucose (GLU), creatinine (Cre), sodium (Na+), potassium (K+), chlorine (Cl-)

URINALYSIS: Yes
- Time schedule for collection of urine: At the end of treatment period and recovery period
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Yes
- Parameters examined: general characters, glucose (Glu), bilirubin (Bil), ketone body (Ket), urinary specific gravity (SG), blood (Bld), pH, urinary protein (Pro), urobilinogen (Uro), nitrite (Nit), white cell (WBC)

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Weekly during the treatment and recovery period
- Dose groups that were examined: All groups in the treatment and recovery period
- Battery of functions tested: pupillary reflex, approach response, touch response, air-righting response, pain perception (Tail pinch), startle response

IMMUNOLOGY: No

OTHER: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Sacrifice date: see schedule of clinical hematology.
All animals were weighed and necropsied. Descriptions of all macroscopical abnormalities were recorded. All animals were anesthetized and killed by exsanguination.

HISTOPATHOLOGY: Yes
The following organs from all animals were weighed before fixation and recorded at scheduled necropsy: Brain, Heart, Liver, Spleen, Lung, kidney, Adrenal glands, Thymus, Testis, Epididymis, Uterus and Ovary.
Relative organ weights were calculated on the basis of the body weight. The terminal body weight was recorded immediately prior to necropsy and the organ to terminal body weight ratios were determined.
Samples of the following tissues and organs were collected from all animals at necropsy and fixed in neutral phosphate buffered formaldehyde solution.
Heart, Lungs, Liver, Spleen, Kidney, Adrenal glands, Pancreas, Oesophagus, Stomach, Small and large intestines, Lymph nodes, Submandibular gland, Thyroid gland, Parathyroid glands, Trachea, Thymus gland, Testes, Ovaries, Epididymides, Uterus, Prostate, Mammary glands, Brain (cerebrum, cerebellum and pons), Pituitary gland, Sciatic nerve with skeletal muscle, Urinary bladder, Spinal cord, Optic nerve, Thoracic aorta, Sternum with bone marrow

Statistics:

A statistical analysis of results of body weight, food consumption, parameters of haematology, blood coagulation and clinical biochemistry and absolute and relative organ weights was performed for each gender by comparing values of dosed with control animals of the main groups using a one-way ANOVA and a post-hoc Dunnet t Test. Pathological data were analyzed descriptively and tissue lesions incidence were analyzed using chi-square Test.
These statistics were performed with SPSS software (p<0.05 was considered as statistically significant).

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

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:

effects observed, non-treatment-related

Description (incidence and severity):

Significantly reduced mean MCV (%) in females and elevated mean ALT (%) in males was noted in the group HD (p< 0.05). No changes of MCV (%) or ALT (%) were observed in the group HDR. Both significantly elevated means of RDW (%) (p< 0.05) and NEUT(%) (p<0.05) in females and significantly reduced mean LYM (%) in females were noted in group HDR. Although some of the differences attained statistical significance (p< 0.05) of the examined parameters, the values of these changes were within historical control data range, an association with treatment was considered unlikely because the changes occurred mainly in group HDR but not group HD. In addition, all hematological changes lacked dose-relationship and therefore were judged to be of no toxicological significance and systematic toxic relevance.
There were no statistically significance changes found in any other measured hematological parameters in treated groups when compared to control group.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

Significantly elevated mean of ALT and Urea in males of the group HD, as well as significantly elevated mean of CHOL in males of the group LD were noted during the treatment period (p< 0.05). The changes of ALT, Urea and CHOL were not seen in the recovery groups.
Significantly elevation of ALP was noted in males of group HDR when compared to control groups (p< 0.05).
However, the values of these changes remained within the range of the historical control data. In addition, all clinical biochemical changes lacked any dose-relationship and therefore were judged to be of no toxicological significance and systematic toxic relevance.
There were no statistically significance changes found in any other measured clinical biochemical parameters in treated groups when compared to control group.

Urinalysis findings:

no effects observed

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

Functional observational battery was conducted weekly. There were marked changes of the urination and defecation noted in treated groups when compared to control group during Day 20 to Day 41.The changes of the measured urination or defecation did not show a trend of continuous consistency or a dose relationship during the whole study. These inter-group differences of the urination and defecation did not show biological significance and therefore judged to be natural variation in measurement during the study.
There was no biologically significant effect observed in any other parameters of functional observational battery in all treated groups during the whole study.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

Absolute organ weights: the mean lung weight of males in group LD was increased (p< 0.05) and the mean adrenal glands weight of males in group LD was decreased (p< 0.01) when compared to group C of the same sex.
Relative organ weights: the mean kidney weight of male in group LD was decreased (p< 0.05) and the mean adrenal glands weight of males in group LD was decreased (p< 0.01) when compared to group C of the same sex.
All these findings were not observed in animals of the group HD. There were no test item-related changes detected in the mean absolute or relative organ weights of treated groups.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

All animal survived up to the scheduled terminal sacrifice of the administration or recovery period.
Immature right testis and epididymis were detected in one male (R2015) of group LD. Discoloured dark parts of lung tissue were observed in one female (R5046 of group CR). All gross lesions noted were single observations. The macroscopic findings were judged to be incidental and not treatment related.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

At the end of treatment period, the relative organ weights of kidney and adrenal glands of males in group LD were decreased significantly when compared with males in control group. There were no corresponding findings of histopathological changes detected in the organs of these animals. The changes of relative organ weight were considered to be not treatment-related and not toxicological significant.
Different degree of focal chronic inflammation observed in liver, kidney and prostate were considered to be spontaneous occurring background pathology findings in this strain of this age. These findings were assumed to be unrelated to treatment. At the end of treatment period, focal necrosis with chronic inflammation in liver was observed in HD group males. A test item related effect could not completely exclude. Since the effect was not observed in the recovery group and was only detected in 2 of 5 animals, the effect was judged to be incidental rather than treatment related.

Histopathological findings: neoplastic:

no effects observed

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 adverse effects were observed

Key result

Critical effects observed:

no

Results:

Table 1. Mean Haematology and Coagulation

Group	Sex	Animal Number	MCV(fL)	RDW(%)	LYM(%)	NEUT(%)
End of Treatment Period
C	M	5	59.3±1.8	13.4±0.2	95.2±1.0	3.8±0.5
	F	5	58.9±1.3	12.6±0.2	95.9±0.8	3.4±0.7
LD	M	5	59.4±1.0	13.6±0.6	94.9±1.0	4.4±0.8
	F	5	57.6±1.2	12.9±0.1	95.4±1.9	3.9±2.0
MD	M	5	58.8±1.5	13.4±0.5	95.4±1.0	3.7±1.1
	F	5	58.3±2.2	12.7±0.5	94.8±0.9	4.3±0.7
HD	M	5	57.8±2.8	13.8±0.3	95.9±0.9	3.4±0.7
	F	5	56.2±0.6*	12.8±0.2	93.8±1.4	5.4±1.7
End of Recovery Period
CR	M	5	56.2±1.6	13.0±0.2	96.5±0.9	3.0±0.7
	F	5	57.0±1.6	12.3±0.4	92.7±1.1	6.2±1.1
HDR	M	5	56.0±1.1	13.7±0.5*	95.3±0.9	3.7±1.1
	F	5	56.8±2.0	12.9±0.4	94.8±1.1*	4.4±1.0*

Data showed as Mean ± SD; *p<0.05 and **p<0.01vs C group with the same gender and period.

Table 2. Mean Clinical Biochemistry

Group	Sex	Animal Number	ALT (U/L)	Urea (mmol/L)	CHOL (mmol/L)	ALP (U/L)
End of Treatment Period
C	M	5	30±5	4.5±0.3	1.14±0.15	217±31
	F	5	27±8	6.1±0.4	1.57±0.37	130±27
LD	M	5	30±4	4.6±0.6	1.46±0.17*	268±88
	F	5	25±2	5.5±0.7	1.51±0.21	138±12
MD	M	5	33±5	4.6±0.5	1.20±0.15	237±39
	F	5	33±6	6.0±1.0	1.77±0.38	138±67
HD	M	5	41±8*	5.5±0.6*	1.32±0.23	254±76
	F	5	35±8	5.9±0.6	1.26±0.16	103±26
End of Recovery Period
CR	M	5	30±3	6.0±0.6	1.36±0.24	190±37
	F	5	19±2	6.9±1.1	1.85±0.32	76±21
HDR	M	5	29±3	6.3±0.2	1.23±0.20	144±17*
	F	5	20±2	7.1±1.2	2.00±0.36	94±16

Data showed as Mean ± SD;*p<0.05 and **p<0.01vs C group with the same gender and period.

Table 3. Mean Absolute Organ Weights (g)

Group	Animal Number	Body weight	Lung	Adrenals
Male
C	5	396.9±21.0	1.560±0.119	0.079±0.008
LD	5	418.6±22.4	1.862±0.262*	0.055±0.009**
MD	5	409.2±40.7	1.617±0.118	0.074±0.007
HD	5	385.4±14.4	1.581±0.090	0.071±0.016
CR	5	449.1±25.3	1.730±0.044	0.069±0.009
HDR	5	457.7±29.7	1.730±0.197	0.081±0.012
Female
C	5	266.2±17.2	1.284±0.119	0.078±0.005
LD	5	258.9±22.9	1.291±0.075	0.077±0.005
MD	5	262.0±15.5	1.233±0.045	0.079±0.013
HD	5	258.1±10.8	1.241±0.101	0.082±0.012
CR	5	284.5±18.3	1.294±0.212	0.076±0.009
HDR	5	279.8±20.4	1.319±0.103	0.082±0.020

Data showed as Mean ± SD; *p<0.05 and **p<0.01 vs C group with the same gender and period; Absolute organ weights of Group C, LD, MD and HD were the end of treatment period, Group CR and HDR were the end of recovery period.

Table 4. Mean Relative Organ Weights to Body Weight (%)

Group	Animal Number	Body weight	Kidney	Adrenals
Male
C	5	396.9±21.0	0.763±0.033	0.020±0.002
LD	5	418.6±22.4	0.666±0.043*	0.013±0.002**
MD	5	409.2±40.7	0.736±0.069	0.018±0.001
HD	5	385.4±14.4	0.743±0.066	0.018±0.004
CR	5	449.1±25.3	0.723±0.047	0.015±0.002
HDR	5	457.7±29.7	0.664±0.018	0.018±0.002
Female
C	5	266.2±17.2	0.754±0.027	0.030±0.002
LD	5	258.9±22.9	0.732±0.070	0.030±0.003
MD	5	262.0±15.5	0.729±0.024	0.030±0.005
HD	5	258.1±10.8	0.775±0.041	0.032±0.004
CR	5	284.5±18.3	0.710±0.021	0.027±0.004
HDR	5	279.8±20.4	0.745±0.090	0.029±0.006

Table 5.: SummaryHistopathologyChanges and Incidence Rate

Organs	Pathological changes	End of Treatment Period				End of Recovery period
		C	LD	MD	HD	CR	HDR
Liver	Chronic inflammatory lesions	1(4/10) 2(2/10)	1(3/10)	1(4/10)	1(6/10) 2(1/10)	1(5/10) 2(1/10)	1 (3/10)
	Fatty vacuoles degeneration	1(5/10) 3(1/10)	1(4/10) 2(2/10)	1(2/10) 2(1/10)	1(1/10) 2(2/10)	1(1/10) 2(1/10)	1 (5/10)
	Focal necrosis	—	—	—	1(2/10)	—	—
Prostate	Interstitial inflammatory cell infiltration	—	—	—	3(1/5)	2(1/5)	2 (1/5) 3 (1/5)

Note: Number “1 - 4” before parentheses means the degree of histopathology changes:1-light;2-slightly;3-midrange;4-severe.

Number in the parentheses means incidence rate of histopathology changes.

“—” means no observed changes or without histopathology examination.

Conclusions:

Based on the data generated in this study, the NOAEL (No Observed Adverse Effect Level) of Aluminium Magnesium Vanadium Oxide is considered to be 1000 mg/kg body weight/day for the 28-day repeated dose oral toxicity study in male and female rats.

Executive summary:

The purpose of this oral toxicity study was to assess the repeated dose toxicity of Aluminium Magnesium Vanadium Oxide, when administered daily to rats by gavage for a period of 28 days. It could provide information to determine the No-Observed Adverse Effect Level (NOAEL) and to select the dose concentrations for chronic studies. The test item Aluminium Magnesium Vanadium Oxide was administered via gavage to groups of 5 male and 5 female Sprague Dawley rats, respectively at dose levels of 0 mg/kg bw/d (group C, corn oil only), 100 mg/kg bw/d (group LD), 300 mg/kg bw/d (group MD) and 1000 mg/kg bw/d (group HD) over a period of 28 days. To observe the reversibility or delayed occurrence of toxic effects, additional 2 satellite groups (group CR of 0 mg/kg bw/d, group HDR of 1000 mg/kg bw/d) of ten animals (five animals per sex) were scheduled for follow-up observation for a 14 days recovery period after the last administration. During the study following observations and examinations were conducted: general observations, body weight, food consumption, functional observational battery, hematology, blood biochemistry and coagulation, urinalyses, organ weight, macropathology and histopathology investigations were undertaken.

Mortality

No animal died during observation period in the present study.

Clinical Signs

No test item related clinical sign was observed in any of the animals during the entire study

Body Weight

The measured mean body weight was increased with the progress of the study in all groups. There was no statistically or biologically significant effect observed on body weight in treated groups when compared with control group.

Food Consumption

There was no statistically or biologically significant effect observed on food consumption in treated groups during the entire study period.

Functional Observational Battery

The observed changes in measured urination or defecation did not show a trend of consistency or a dose relationship. Therefore, the differences were judged to be incidental rather than treatment related.

Hematology and Blood Coagulation

Significantly reduced mean MCV (%) in females and elevated mean ALT (%) in males were noted in the HD group. No changes in MCV (%) or ALT (%) were observed in the recovery group. Significantly elevated mean of RDW (%) and NEUT(%) in females and significantly reduced mean LYM (%) in females were noted in group HDR. Although some of the differences attained statistical significance of the examined parameters, the values of these changes were within the historical control data range. An association with treatment was considered unlikely because the changes occurred mainly in group HDR but not group HD. In addition, all hematological changes were considered lacked dose-relationship and therefore considered of no toxicological significance and systematic toxic relevance. Blood coagulation was not affected by test item in both males and females.

Clinical Biochemistry

Significantly elevated values of ALT and Urea in males of the group HD, as well as a significantly elevated value of CHOL in males of the group LD were noted during the treatment period. The changes of ALT, Urea and CHOL were not observed in the recovery groups. Significantly elevated values of ALP were noted in males of group HDR when compared to control group. However, the values of these changes remained within the ranges of the historical control data. In addition, all clinical biochemical changes were considered lacking dose-relationship and therefore considered to be incidental rather than treatment related.

Urinalysis

There were no statistically or biological significant changes observed compared to those in the control group in any examination parameters of treated group.

Pathology

All animal survived until terminal sacrifice terminal sacrifice of the exposure or recovery period. Undeveloped right testis and epididymis were found in male R2015 of group LD. Discoloured dark part of lung tissue was observed in female R5046 of group CR. All gross lesions noted were single observations. The macroscopic findings were considered to be incidental and not treatment related.

Organ Weight

Absolute organ weights: the mean lung weight of males in group LD was increased and the mean adrenal glands weight of males in group LD was decreased compared to group C of the same sex. Relative organ weights: the mean kidney weight of males in group LD was decreased and the mean adrenal glands weight of males in group LD was decreased when compared to group C of the same sex. All these findings were not observed in the high dose group. There were no test item-related changes detected in the mean absolute or relative organ weights of treated groups.

Histopathology

Different degrees of focal chronic inflammation observed in liver, kidney and prostate were considered to be spontaneously occurring background pathology findings in this strain of this age. These findings were assumed to be unrelated to treatment. At the end of treatment period, focal necrosis with chronic inflammation in liver was observed in HD group males (2/5 animals).

Discussion

Minimal to slight focal necrosis combined with chronic inflammation was observed in the liver of 2 of 5 males of the high dose group. All biochemical parameters representing liver function including alanine aminotransferase (AST), serum total bilirubin (TB), direct bilirubin (DB), urinary bilirubin and total cholesterol were within historical control data. Additionally the effect was not observed in the recovery group. Therefore, the effect was judged to be incidental rather than treatment related.

Conclusion

Based on the data generated in this study, the NOAEL (No Observed Adverse Effect Level) of Aluminium Magnesium Vanadium Oxide is considered to be 1000 mg/kg body weight/day for the 28-day repeated dose oral toxicity study in male and female rats.

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 Nov 2014 - 25 Jan 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

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)

Remarks:

National (CFDA) GLP certificate

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:Experimental Animal Center of Academy of Military Medical Sciences
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 5-7 weeks
- Weight at study initiation: Male 188.8 ± 8.4 g, female 165.7 ± 7.4 g
- Fasting period before study: Non- fasted animals at start of dosing
- Housing: The rats were housed in groups of 5 animals of one sex per cage.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Animals were quarantined for an acclimatization period of seven days.

DETAILS OF FOOD AND WATER QUALITY:
The SPF mouse/rat maintenance diet was provided by the Experimental Animal Center of Academy of Military Medical Sciences (license No: SCXK (Army) 2012-0003). The analyses report of nutrients and contaminants were provided by diet manufacturers in every quarter of year.
The results of representative reports showed that the nutrients and pollutants level was in compliance with national standards (GB 14924.2-2001 Laboratory animals-Hygienic standard for formula feeds and GB 14924.3-2010 Laboratory animals-Nutrients for formula feeds) without interference or impact on the study.
Animal drinking water was filter-sterilized prepared by water filtered machine. Rats had free access to water. The microbes and other indicators of drinking water were analyzed twice a year. The results of representative water quality analytical reports showed that the microbes and pollutant contaminants level was in compliance with national standard (GB5749-2006 Standards for drinking water quality) without interference or impact on study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25°C
- Humidity (%): 40-70%
- Air changes (per hr): Not specified
- Photoperiod (hrs dark / hrs light): day/nightcycle was 12 hours (12 hours light from 06:00-18:00, 12 hours dark from 18:00-06:00)

IN-LIFE DATES: From: 21 Nov 2014 To:25 Mar 2015

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test item formulations were freshly prepared every day immediately prior to application. Appropriate amount of the test item was weighted into a tared glass vial on a suitable precision balance and the vehicle (corn oil) was added to obtain the designed final concentration of the test item formulations. Homogeneity of the test item in the vehicle was maintained by vortexing the prepared suspension thoroughly before dose administration.
The vehicle (corn oil) was used as the control formulation.


VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil is frequently used as a suitable vehicle in animal experiment when the solubility of test article is low in water. Moreover, coin oil has been proposed as a suitable vehicle by the OECD (OECD Guidelines for Testing of Chemicals: 408 repeated dose 90-Day oral toxicity study in rodents, Sep 1998). Sponsor has agreed to use corn oil as vehicle in this study.
- Concentration in vehicle: 25, 75 and 250 mg/mL
- Amount of vehicle (if gavage): 0.4 mL/100 g bw
- Lot/batch no. (if required): QS130302011011
- Purity:according to the nactional guidance on maize oil (GB 19111-2003)

Analytical verification of doses or concentrations:

no

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.

Duration of treatment / exposure:

90 days

Frequency of treatment:

Once daily, continuously

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Low Dose(LD) group

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Medium Dose (MD) group

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

High dose(HD) group
high dose with recovery period (HDR) group

No. of animals per sex per dose:

10 aminals per sex per dose in control (C), LD, MD and HD groups
5 animals per sex per dose in satellite group: control recovery (CR) and HDR group

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose levels were selected based on the results of a GLP 28-day repeated dose oral toxicity study in Sprague Dawley rats. The NOAEL of 28-day repeated dose oral toxicity study was 1000 mg/kg bw.

- Post-exposure recovery period in satellite groups: 28 days

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once a day during the treatment and recovery periods
- Cage side observations included: mortality/viability

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly during the treatment and recovery periods.

BODY WEIGHT: Yes
- Time schedule for examinations: Once by animal receiving; once before experimental grouping; weekly during the treatmentand recovery period and once before necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Weekly during the treatment and recovery periods
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: before administration and at the end of treatment/recovery period.
- Dose groups that were examined: All animals of C, LD, MD, HD, CR and HDR groups before administration; at the end of treatment/recovery period, all animals of C and HD groups are checked firstly, other groups of animals are subsequent checked when C and HD groups have ophthalmologic findings.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of treatment period and recovery periods
- Anaesthetic used for blood collection: Yes
- Animals fasted: Yes
- How many animals: all
- Parameters examined: red blood cell (RBC), hemoglobin (HGB), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelet (PLT), platelet distribution width (PDW), hematocrit (HCT), mean corpuscular volume (MCV), red cell distribution width (RDW), mean platelet volume (MPV), white blood cell (WBC) and classification.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of treatment period and recovery period
- Animals fasted: Yes
- How many animals:all
- Parameters examined: aspartate amino transferase (AST), alanine amino transterase (ALT), alkaline phosphatase (ALP), creatine kinase (CK), total protein (TP), albumin (ALB), total bilirubin (TB), direct bilirubin (DB), triglycerides (TG), urea (Urea), total cholesterol (TC), glucose (GLU), creatinine (Cre), sodium (Na+), potassium (K+), chlorine (Cl-)

URINALYSIS: Yes
- Time schedule for collection of urine: At the end of treatment period and recovery period
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Yes
- Parameters examined: general characters, glucose (Glu), bilirubin (Bil), ketone body (Ket), urinary specific gravity (SG), blood (Bld), pH, urinary protein (Pro), urobilinogen (Uro), nitrite (Nit), white cell (WBC)

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Weekly during the treatment and recovery period
- Dose groups that were examined: All groups in the treatment and recovery period
- Battery of functions tested: pupillary reflex, approach response, touch response, air-righting response, pain perception (Tail pinch), startle response

IMMUNOLOGY: No

OTHER: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Sacrifice date: At the terminal of treatment period (26-Mar-2015) and recovery period (23-Apr-2015).
All animals were weighed and necropsied. Descriptions of all macroscopical abnormalities were recorded. All animals were anesthetized and killed by exsanguination.
The following organs from all animals were weighed before fixation and recorded at scheduled necropsy: Brain, Heart, Liver, Spleen, Lung, kidney, Adrenal glands, Thymus, Testis, Epididymis, Uterus and Ovary.
Relative organ weights were calculated on the basis of the body weight. The terminal body weight was recorded immediately prior to necropsy and the organ to terminal body weight ratios were determined.

HISTOPATHOLOGY: Yes
Samples of the following tissues and organs were collected from all animals at necropsy and fixed in neutral phosphate buffered formaldehyde solution.
Heart, Lungs, Liver, Spleen, Kidney, Adrenal glands, Pancreas, Oesophagus, Stomach, Small and large intestines, Lymph nodes, Submandibular gland, Thyroid gland, Parathyroid glands, Trachea, Thymus gland, Testes, Ovaries, Epididymides, Uterus, Prostate, Mammary glands, Brain (cerebrum, cerebellum and pons), Pituitary gland, Sciatic nerve with skeletal muscle, Urinary bladder, Spinal cord, Optic nerve, Thoracic aorta, Sternum with bone marrow.

Statistics:

A statistical analysis of results of body weight, food consumption, parameters of hematology, blood coagulation and clinical biochemistry and absolute and relative organ weights was performed for each gender by comparing values of dosed with control animals of the main groups using a one-way ANOVA and a post-hoc Dunnet t Test. Pathological data were analyzed descriptively and tissue lesions incidence were analyzed using chi-square Test. These statistics were performed with SPSS software (p<0.05 was considered as statistically significant).

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

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:

no effects observed

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Dose-dependent reduction of MPV (fL) (p<0.05) was observed in males, while there were no changes of PLT (109/L) noted when compared to the control group. Dose-dependent reduction of MCV (fL) was noted in females (p<0.05), while there were no changes of RBC (1012/L) observed when compared to the control group. Significantly reduced means of RDW (%) (p<0.05) in females of group LD, and significantly reduced means of PDW (%) and EOS (%) (p<0.05) in females of group HD were noted, respectively. Although some of the differences attained statistical significance (p< 0.05) of the examined parameters, the values of these changes were within historical control data range.
There were no statistically significance changes found in any other measured hematological parameters in treated groups when compared to control group.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

Dose-dependent reduction of TP (g/L) (p<0.01) was observed in males when compared to the control group. Significantly elevated means of Urea (mmol/L) (p<0.05) was noted in males and females of the group HD, but there was no change of CREA (μmol/L) observed. Significantly elevated means of ALT (U/L) (p<0.05) was noted in males of group MD, and significantly reduced means of ALB (g/L) (p<0.05) and GLU (mmol/L) (p<0.01) were noted in males of group HD and HDR, respectively. Significantly reduction of AST (U/L) (p<0.05) was noted in females of group HD. However, the values of these changes remained within the ranges of the historical control data.
There were no statistically significance changes found in any other measured clinical biochemical parameters in treated groups when compared to control group.

Urinalysis findings:

effects observed, non-treatment-related

Description (incidence and severity):

Significantly changes of BIL (p<0.05) was noted in females of group LD. However, there were no significantly changes in TBIL and D-BIL in the clinical biochemistry and therefore the changes of BIL was judged to be of no toxicological significance. There were no statistically or biological significant changes compared to those in the control group in any other examination parameter of treated groups.

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

Functional observational battery was conducted weekly. There were marked changes of the urination and defecation noted in the treatment groups when compared to control group. The changes of the measured urination or defecation did not show a trend of continuous consistency or a dose relationship during the whole study. These inter-group differences of the urination and defecation did not show biological significance and therefore were judged to be variations in measurement during the study.
There was no biologically significant effect observed in any other parameters of functional observational battery in all treated groups during the whole study.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

Absolute organ weights: the mean thymus gland weight of males in group HD and the mean liver weight of females in group LD was decreased (p< 0.05) when compared to group C of the same sex. The changes only were judged to be have no toxicological significance.
Relative organ weights: there was no significantly change noted in any of the dosed group when compared to group C of the same sex.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

All animal survived up to scheduled terminal sacrifice of the administration or recovery period.
Stomach, spleen and abdominal adhesions was observed in one female (R1016) of control group. Nephrohydrosis was found in one male (R4063) of group HD. Visible hard mass of right kidney was observed in one female (R4080) of group HD. All gross lesions noted were single observations. The macroscopic findings were judged to be incidental and not treatment related.
No other changes were detected associated with the test item.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

Different degree of focal chronic inflammation observed in heart and kidney in control and HD groups. Prostate chronic inflammation was noted in few animals in control, HD and HDR group. These findings were assumed to be unrelated to treatment and judged to be spontaneous occurring background pathology findings in this strain of this age.
At the end of treatment period, local proliferation and focal inflammation lesions focal inflammation in kidney in individual animal was judged to be incidental rather than treatment-related.
There were no test item treated related histopathological changes found in the 90-day repeated dose oral toxicity with Aluminium Magnesium Vanadium Oxide.

Histopathological findings: neoplastic:

no effects observed

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 adverse effects observed

Key result

Critical effects observed:

no

Results:

Table 1. Mean Haematology and Coagulation

Group	Sex	Animal Number	MCV(fL)	RDW (%)	MPV (fL)	PDW (%)
End of Treatment Period
C	M	10	53.1±1.6	13.4±0.5	3.9±0.4	14.8±0.7
	F	10	55.9±1.6	13.1±1.1	4.1±0.5	15.2±0.7
LD	M	10	52.3±2.5	13.1±0.3	3.6±0.5	14.8±0.6
	F	10	54.4±2.0	12.5±0.3*	3.8±0.5	14.9±0.6
MD	M	10	50.7±2.0	13.4±0.3	3.4±0.4*	14.4±0.4
	F	10	54.3±0.9	12.7±0.4	3.8±0.5	14.9±0.6
HD	M	10	51.8±2.3	13.4±0.4	3.2±0.4**	14.3±0.4
	F	10	53.8±1.1*	12.7±0.2	3.1±0.5**	14.3±0.6**
End of Recovery Period
CR	M	5	51.5±1.5	13.5±0.3	3.7±0.4	14.5±0.3
	F	5	55.6±2.8	12.7±0.1	4.1±0.5	15.2±0.6
HDR	M	5	52.1±2.2	13.9±0.2	3.8±0.4	14.9±0.5
	F	5	54.2±1.4	13.0±0.2	3.8±0.3	15.0±0.8

Data showed as Mean ± SD;*p<0.05 and **p<0.01vs C group with the same gender and period.

Table 2. MeanHaematology and Coagulation

Group	Sex	Animal Number	EOS(%)
End of Treatment Period
C	M	10	0.4±0.2
	F	10	0.8±0.7
LD	M	10	0.5±0.2
	F	10	0.7±0.4
MD	M	10	0.7±0.4
	F	10	0.8±0.4
HD	M	10	0.6±0.3
	F	10	0.5±0.2
End of Recovery Period
CR	M	5	0.7±0.5
	F	5	1.3±0.5
HDR	M	5	0.6±0.3
	F	5	0.5±0.2*

Data showed as Mean ± SD;*p<0.05 and **p<0.01vs C group with the same gender and period

Table 3. Mean Clinical Biochemistry

Group	Sex	Animal Number	ALT (U/L)	AST (U/L)	TP (g/L)	ALB (g/L)	GLU (mmol/L)	Urea (mmol/L)
End of Treatment Period
C	M	10	31±4	82±18	58.8±1.2	21.9±0.9	7.97±0.39	4.4±0.8
	F	10	31±5	84±8	64.4±3.3	25.9±2.8	8.28±0.72	4.7±0.6
LD	M	10	36±6	75±16	57.8±2.1	21.9±0.8	7.50±0.74	4.2±0.3
	F	10	35±8	76±8	65.2±3.4	26.2±2.7	8.11±1.16	5.2±1.1
MD	M	10	39±6*	78±9	55.8±2.0**	21.3±0.6	7.82±1.11	5.0±0.6
	F	10	29±7	80±14	64.2±3.8	26.7±2.0	8.19±0.97	5.4±1.1
HD	M	10	38±11	75±9	55.3±2.8**	21.0±0.7*	7.64±0.95	6.3±1.1**
	F	10	28±6	68±22*	60.8±4.5	24.9±2.1	7.85±0.57	5.9±1.1*
End of Recovery Period
CR	M	5	34±3	120±17	61.1±2.4	21.2±0.5	9.26±0.60	5.3±0.3
	F	5	25±3	74±10	68.0±5.2	26.4±2.1	8.56±1.06	5.7±0.6
HDR	M	5	33±4	100±19	59.4±3.1	20.6±1.1	7.73±0.59**	5.2±0.9
	F	5	21±2	65±9	68.8±5.0	26.5±2.2	7.79±0.85	5.7±0.3

Data showed as Mean ± SD;*p<0.05 and **p<0.01vs C group with the same gender and period.

Table 4. MeanUrinalyses

Group	Sex	N	Bil
C	M	5	−: 1/5
			+: 4/5
	F	5	−:2/5
			+: 1/5
			2+: 1/5
			3+: 1/5
LD	M	5	−:2/5
			+: 1/5
			3+: 2/5
	F	5
			−: 2/5
			+: 3/5
MD	M	5	−: 2/5*
			2+:3/5
	F	5	−: 3/5
			+: 1/5
			3+:1/5

*p<0.05 and **p<0.01vs C group with the same gender and period.

Table 5. Mean Absolute Organ Weights (g)

Group		Animal Number		Body weight		Thymus		Liver
Male
C		10		593.3±50.9		0.416±0.046		14.215±1.636
LD		10		594.2±64.3		0.375±0.098		14.185±2.172
MD		10		580.7±38.9		0.384±0.079		14.003±1.359
HD		10		563.3±30.0		0.331±0.075*		13.127±1.803
CR		5		691.4±90.5		0.385±0.066		16.408±3.552
HDR		5		618.4±64.4		0.354±0.109		14.743±2.083
Female
C		10		334.8±31.7		0.317±0.098		9.234±0.928
LD		10		321.2±32.5		0.312±0.076		8.400±0.590*
MD		10		316.7±34.4		0.318±0.109		8.666±0.791
HD		10		320.1±18.0		0.317±0.078		8.758±0.575
CR		5		334.9±29.5		0.314±0.032		8.849±1.391
HDR		5		324.9±26.2		0.299±0.060		8.593±0.824

Data showed as Mean ± SD;*p<0.05 and **p<0.01vs C group with the same gender and period; Absolute organ weights of Group C, LD, MD and HD were the end of treatment period, Group CR and HDR were the end of recovery period.

Table 6. SummaryHistopathologyChanges and Incidence Rate

 

Organs	Pathological changes	End of Treatment Period				End of Recovery Period
		C	LD	MD	HD	CR	HDR
Lung	Chronic bronchioloalveolar inflammation	2 (4/20) 3 (5/20)	—	—	1 (3/20) 2 (5/20) 3 (1/20)	1 (4/10) 2 (3/10) 3 (1/10)	1 (6/10)
Prostate	Interstitial inflammatory cell infiltration	1 (1/10) 2 (2/10)	—	—	1 (1/10) 2 (1/10)	3 (1/5)	1 (1/5)

Note: Number “1 - 4” before parentheses means the degree of histopathology changes:1-light;2-slightly;3-midrange;4-severe.

Number in the parentheses means incidence rate of histopathology changes.

“—” means no observed changes or without histopathology examination.

Conclusions:

In a reliable study conducted according to OECD 408 and in compliance with GLP, administration of the test item at a dose of 1000 mg/kg bw/day did not reveal adverse effects. Thus, the No Observed Adverse Effect Level (NOAEL) of Aluminium Magnesium Vanadium Oxide was determined to be 1000 mg/kg body weight/day for the 90-day repeated dose oral toxicity study in male and female rats.

Executive summary:

The purpose of this oral toxicity study was to assess the repeated dose toxicity of Aluminium Magnesium Vanadium Oxide, when administered daily to rats by gavage for a period of 90 days. It could provide information to determine the No-Observed Adverse Effect Level (NOAEL) and to select the dose concentrations for chronic studies.

The test item Aluminium Magnesium Vanadium Oxide was administered via gavage to groups of 10 male and 10 female Sprague Dawley rats, respectively at dose levels of 0 mg/kg bw/d (group C, corn oil only), 100 mg/kg bw/d (group LD), 300 mg/kg bw/d (group MD) and 1000 mg/kg bw/d (group HD) over a period of 90 days. To observe the reversibility or delayed occurrence of toxic effects, additional 2 satellite groups (group CR of 0 mg/kg bw/d, group HDR of 1000 mg/kg bw/d) of ten animals (five animals per sex) were scheduled for follow-up observation for a 28 days recovery period after the last administration.

During the study following observations and examinations were conducted: general observations, body weight, food consumption, functional observational battery, ophthalmologic examination, hematology, blood biochemistry and coagulation, urinalyses, organ weight, macropathology and histopathology investigations were undertaken.

Mortality

No animal died during observation period in the present study.

Clinical Signs

No test item related clinical sign was observed in any of the animals during the entire study

Body Weight

The measured mean body weight was increased with the progress of the study in all groups. There was no statistically or biologically significant effect observed on body weight in treated groups when compared with control group.

Food Consumption

The slight differences of food consumption among the groups were considered as incidental fluctuations of food intake due to the absence of dose relationship. There was no statistically or biologically significant effect observed on food consumption in treated groups during the entire study period.

Ophthalmologic Examination

No treatment related findings were observed in ophthalmologic examination in all groups at the end of treatment and recovery periods.

Functional Observational Battery

The observed changes in measured urination or defecation did not show a trend of consistency or a dose relationship. Therefore, the differences were judged to be incidental rather than treatment related.

Hematology and Blood Coagulation

Dose-dependent reduction of MPV (fL) (p<0.05) was observed in males, but there was no changes of PLT (10⁹/L) noted when compared to the control group. Dose-dependent reduction of MCV (fL) was noted in females (p<0.05), but there was no changes of RBC (10¹²/L) observed when compared to the control group. Significantly reduced means of RDW (%) (p<0.05) in females of group LD, and significantly reduced means of PDW (%) and EOS (%) (p<0.05) in females of group HD were noted, respectively. Although some of the differences attained statistical significance (p< 0.05) of the examined parameters, the values of these changes were within historical control data range.

Blood coagulation was not affected by test item in both males and females.

Clinical Biochemistry

Dose-dependent reduction of TP (g/L) (p<0.01) was observed in males when compared to the control group. Significantly elevated means of Urea (mmol/L) (p<0.05) was noted in males and females of the group HD, but there was no change of CREA (μmol/L) observed. Significantly elevated means of ALT (U/L) (p<0.05) was noted in males of group MD, and significantly reduced means of ALB (g/L) (p<0.05), AST (U/L) (p<0.05)as well as GLU (mmol/L) (p<0.01)were noted in males of group HD.

However, the values of these changes remained within the ranges of the historical control data. In addition, all clinical biochemical changes were considered lacking dose-relationship and therefore considered to be incidental rather than treatment related.

Urinalysis

Significantly changes of BIL (p<0.05) was noted in females of group LD. However, there were no significantly changes in TBIL and D-BIL in the clinical biochemistry and therefore the changes of BIL was judged to be of no toxicological significance. There were no statistically or biological significant changes compared to those in the control group in any other examination parameter of treated groups.

Pathology

All animal survived until terminal sacrifice terminal sacrifice of the exposure or recovery period. Stomach, spleen and abdominal adhesions was observed in one female (R1016) of control group. Nephrohydrosis were detected in one male (R4063) of group HD. Visible hard mass of right kidney was observed in one female (R4080) of group HD. All gross lesions noted were single observations. The macroscopic findings were judged to be incidental and not treatment related.

Organ Weight

The organ weights were recorded from the surviving animals and relative organ weights in relation to body weight were calculated. Absolute organ weights: the mean thymus gland weight of males in group HD and the mean liver weight of females in group LD was decreased (p< 0.05) when compared to group C of the same sex. The changes only were judged to be have no toxicological significance. Relative organ weights: there was no significantly change noted in any of the dosed group when compared to group C of the same sex.

Histopathology

Different degree of focal chronic inflammation observed in heart and kidney in control and HD groups. Prostate chronic inflammation was noted in few animals in control, HD and HDR group. These findings were assumed to be unrelated to treatment and judged to be spontaneous occurring background pathology findings in this strain of this age. At the end of treatment period, local proliferation and focal inflammation lesions focal inflammation in kidney in individual animal was judged to be incidental rather than treatment-related. No treatment related microscopic findings were observed in kidney of other animals of all groups at the end of treatment and recovery periods. There were no test item treated related histopathological changes found in the 90-day repeated dose oral toxicity with aluminium magnesium vanadium oxide.

Discussion

No treatment related findings were observed in all groups at the end of treatment and recovery periods, including mortality, clinical signs, body weight, food consumption, ophthalmologic examination, functional observational battery, hematology and blood coagulation, clinical biochemistry, urinalysis, pathology and organ weight. Different degree of focal chronic inflammation observed in heart and kidney in control and HD groups. Prostate chronic inflammation was noted in few animals in control, HD and HDR group. These findings were assumed to be unrelated to treatment and judged to be spontaneous occurring background pathology findings in this strain of this age. At the end of treatment period, local proliferation and focal inflammation lesions focal inflammation in kidney in individual animal was judged to be incidental rather than treatment-related. No treatment related microscopic findings were observed in kidney of other animals of all groups at the end of treatment and recovery periods. There were no test item treated related histopathological changes found in the 90-day repeated dose oral toxicity with aluminium magnesium vanadium oxide.

Conclusion

Based on the data generated in this study, the NOAEL (No Observed Adverse Effect Level) of Aluminium Magnesium Vanadium Oxide is considered to be 1000 mg/kg body weight/day for the 90-Day repeated dose oral toxicity study in male and female rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 1) from the test substance. The study is sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No. 1907/2006.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Two repeated dose oral toxicity studies were performed for aluminium magnesium vanadium oxide according to OECD Guidelines for Testing of Chemicals 408, “Repeated Dose 90-Day Oral Toxicity Study in Rodents (1998)” and 407, “Repeated Dose 28-Day Oral Toxicity Study in Rodents (2008)”.

In the 90-day study, ten Sprague Dawley rats/sex/dose were administered 0 (control, C), 100 (low dose group, LD), 300 (medium group, MD) and 1000 (high dose group, HD) mg/kg bw/day aluminium magnesium vanadium oxide in corn oil once daily for 90 days via gavage. In addition, a satellite control and high-dose group of 10 rats/dose /sex was treated in parallel, with a 28-day post-exposure recovery period to assess the reversibility of treatment-related effects.

There was no mortality during the study period. No toxicologically relevant clinical signs, ophthalmologic examination findings and Functional Observational Battery (FOB) findings were observed. The body weight, body weight gain and food consumption was comparable between the control and treatment groups (main and satellite groups).

Dose-dependent reduction of MPV (p<0.05) was observed in males, but there was no changes of PLT noted when compared to the control group. Dose-dependent reduction of MCV was noted in females (p<0.05), but there was no changes of RBC observed when compared to the control group. Significantly reduced means of RDW (p<0.05) in females of group LD, and significantly reduced means of PDW and EOS (p<0.05) in females of group HD were noted, respectively. Although some of the differences attained statistical significance (p< 0.05) of the examined parameters, the values of these changes were within historical control data range. Blood coagulation was not affected by test item in both males and females. Dose-dependent reduction of TP (p<0.01) was observed in males when compared to the control group. Significantly elevated means of Urea (p<0.05) was noted in males and females of the group HD, but there was no change of CREA observed. Significantly elevated means of ALT (p<0.05) was noted in males of group MD, and significantly reduced means of ALB (p<0.05), AST (p<0.05)as well as GLU (p<0.01)were noted in males of group HD. However, the values of these changes remained within the ranges of the historical control data. In addition, all clinical biochemical changes were considered lacking dose-relationship and therefore considered to be incidental rather than treatment related. Significantly changes of BIL (p<0.05) was noted in females of group LD. However, there were no significant changes in TBIL and D-BIL in the clinical biochemistry and therefore the changes of BIL was judged to be of no toxicological significance. There were no statistically or biological significant changes compared to those in the control group in any other examination parameter of treated groups.

All animal survived until terminal sacrifice of the exposure or recovery period. Stomach, spleen and abdominal adhesions was observed in one female (R1016) of control group. Nephrohydrosis were detected in one male (R4063) of group HD. Visible hard mass of right kidney was observed in one female (R4080) of group HD. All gross lesions noted were single observations. The macroscopic findings were judged to be incidental and not treatment related. The organ weights were recorded from the surviving animals and relative organ weights in relation to body weight were calculated. Absolute organ weights: the mean thymus gland weight of males in group HD and the mean liver weight of females in group LD was decreased (p< 0.05) when compared to group C of the same sex. Relative organ weights: there was no significantly change noted in any of the dosed group when compared to group C of the same sex. Different degree of focal chronic inflammation observed in heart and kidney. Prostate chronic inflammation was noted in few animals at the end of treatment and recovery groups. These findings were assumed to be unrelated to treatment and judged to be spontaneous occurring background pathology findings in this strain of this age. At the end of treatment period, local proliferation and focal inflammation lesions focal inflammation in kidney in individual animal was judged to be incidental rather than treatment-related. No treatment related microscopic findings were observed in kidney of other animals of all groups at the end of treatment and recovery periods. There were no test item treated related histopathological changes found in the 90-day repeated dose oral toxicity with aluminium magnesium vanadium oxide.

In the subacute 28-day study, five Sprague Dawley rats/sex/dose were administered 0 (control, C), 100 (low dose group, LD), 300 (medium group, MD) and 1000 (high dose group, HD) mg/kg bw/day aluminium magnesium vanadium oxide in corn oil once daily for 28 days via gavage. In addition, a satellite control and high-dose group of 5 rats/dose /sex was treated in parallel, with a 14-day post-exposure recovery period to assess the reversibility of treatment-related effects.

There was no mortality during the study period. No toxicologically relevant clinical signs and functional observational battery (FOB) findings were observed. The body weight, body weight gain and food consumption was comparable between the control and treatment groups (main and satellite groups).

Significantly reduced mean MCV in females and elevated mean ALT in males were noted in the HD group. No changes in MCV or ALT were observed in the recovery group. Significantly elevated mean of RDW and NEUT in females and significantly reduced mean LYM in females were noted in group HDR. Although some of the differences attained statistical significance of the examined parameters, the values of these changes were within the historical control data range. An association with treatment was considered unlikely because the changes occurred mainly in group HDR but not group HD. In addition, all hematological changes were considered lacked dose-relationship and therefore considered of no toxicological significance and systematic toxic relevance.Significantly elevated values of ALT and Urea in males of the group HD, as well as a significantly elevated values of CHOL in males of the group LD were noted during the treatment period. The changes of ALT, Urea and CHOL were not observed in the recovery groups. Significantly elevated values of ALP were noted in males of group HDR when compared to control group. However, the values of these changes remained within the ranges of the historical control data. There were no statistically or biological significant changes observed compared to those in the control group in any examination parameters of treated group.

All animal survived until terminal sacrifice of the exposure or recovery period. The absolute organ weight of lung of males in group LD was increased and the mean adrenal glands weight of males in group LD was decreased compared to group C of the same sex. The relative organ weight of kidney of males in group LD was decreased and the mean adrenal glands weight of males in group LD was decreased when compared to group C of the same sex. All these findings were not observed in the high dose group. There were no test item-related changes detected in the mean absolute or relative organ weights of treated groups. Undeveloped right testis and epididymis were found in male R2015 of group LD. Discoloured dark part of lung tissue was observed in female R5046 of group CR. All gross lesions noted were single observations. The macroscopic findings were considered to be incidental and not treatment related. Different degree of focal chronic inflammation observed in liver, kidney and prostate were considered to be spontaneous occurring background pathology findings in this strain of this age. These findings were assumed to be unrelated to treatment. Focal necrosis with chronic inflammation in liver was observed in HD group males. A test item related effect could not completely exclude but since the effect was not observed in the recovery group and was only detected in 2 of 5 animals, the effect was judged to be incidental rather than treatment related.

 

Based on the data generated in the both studies, the NOAEL (No Observed Adverse Effect Level) of aluminium magnesium vanadium oxide is considered to be greater than 1000 mg/kg body weight/day for the 90-Day repeated dose oral toxicity study in male and female rats.

Justification for classification or non-classification

Based on the available data on repeated dose toxicity indicate that the Aluminium Magnesium Vanadium Oxide dose not meet the critiria for classification according to Regulation(EC) 1272/2008.