Aluminum magnesium vanadium oxide

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproduction/Developmental Toxicity Screening Test (OECD 421), rat: 1000 mg/kg body weight/day (m/f)

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 Oct 2014 - 16 Mar 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

1995

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 nulliparous and non-pregnant: yes
- Age at study initiation: (P) x 10-11wks
- Weight at study initiation: (P) Males: 317.9-359.8 g; Females: 218.7-270.1 g
- Fasting period before study: No
- Housing:The rats were housed five of one sex per cage at treatment before mating and after mating (only males). The pregnant animals were single-housed after mated. Animals were mated in the ratio of 1:1 (Male to Female) during the mating period.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25°C
- Humidity (%): 40-70%
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light):12 hrs dark/12 hrs light
IN-LIFE DATES: from 22 Dec 2014 to 13 Feb 2015

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:The test item formulations were freshly prepared every day immediately prior to application.
Appropriate amounts of the test item were 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 administration. The vehicle (corn oil) was also used as the control item.

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, corn oil has been proposed as a suitable vehicle by the OECD (OECD Guidelines for Testing of Chemicals 421Reproduction/Developmental Toxicity Screening Test (Adopted 24, July, 1995). Sponsor has agreed to use corn oil as vehicle in this study.
- Concentration in vehicle: Not specified
- Amount of vehicle (if gavage): 0.4 mL /100g body weight
- Lot/batch no. (if required): QS130302011011
- Purity: according to the nactional guidance on maize oil (GB 19111-2003)

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- Further matings after two unsuccessful attempts: not specified
- After successful mating each pregnant female was caged (how): The pregnant animals were single-housed after mated
- Any other deviations from standard protocol: No

Analytical verification of doses or concentrations:

no

Remarks:

No analytical need for poor soluble inorganic substance

Duration of treatment / exposure:

approximately 54 days for female (day14 pre-mating to day 4 post-partum )
28 days for male (day14 pre-mating and day14 of mating )

Frequency of treatment:

daily

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:

12

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Toxicological Information: LC50 (inhalation) > 5 mg/L; LD50 (dermal) > 2000 mg/kg body weight; LD50 (oral) > 2000 mg/kg body weight; no signs of toxicity at acute sublethal concentration;

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once daily
- Cage side observations included: mortality/viability

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed at receiving, grouping, first day of dosing and weekly thereafter, and at termination. During pregnancy, females were weighed every 3 days and within 24 hours after parturition (day 0 or 1 post-partum) and day 4 post-partum.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

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

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities;

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals were killed at day 28 of dosing
- Maternal animals: Pregnant females animals were killed at day 4 post-partum, and non-pregnant females were killed 24-26 days after the last day of the mating period.

GROSS NECROPSY
- At the time of sacrifice or death during the study, the adult animals were examined macroscopically for any abnormalities or pathological changes. Special attention was paid to the organs of the reproductive system. The pregnancy status of the females was ascertained. The number of implantation sites, the counting of corpora lutea, and the number of absorbed fetus and dead fetus were recorded. Uteri that appear non-gravid should be further examined by 8% ammonium sulphide staining for 20 min to confirm the non-pregnant status with or without implantation.

HISTOPATHOLOGY / ORGAN WEIGHTS
Detailed histological examination was performed on the ovaries, uterus, testes and epididymides of the animals of the highest dose group and the control group. Examinations were not extended to the animals of other dosage groups, because no changes were seen in the highest dose group. The testes and epididymides of male adult and ovaries and uterus of female adult animals were weighed. The absolute organ weights and the relative organ weights on the basis of the body weight were calculated. The ovaries, uterus, testes, epididymides, accessory sex organs and all organs showing macroscopic lesions of all adult animals were preserved using Bouin’s fixative.

Postmortem examinations (offspring):

SACRIFICE
Dead pups and pups killed at day 4 post-partum were examined externally for gross abnormalities.

Statistics:

The numerical data obtained in body weight, food consumption, organ weight and relative organ weight were subjected to multiple comparison tests for statistical significance of difference. The data were analyzed for homogeneity of variance and the homogeneous data were subjected to one-way analysis of variance. If significant differences were observed among groups, group mean difference between the control group and the dose group was subjected to multiple comparison test by the Dunnet t test (for homogeneous data). The enumeration data were analyzed using chi-square Test. These statistics were performed with SPSS 18.0 software (p<0.05 was considered as statistically significant)
Individual animal data were provided. Additionally, all data were summarized in tabular form, showing for each test group the number of animals at the start of the test, the number of animals found dead during the test or killed for human reasons, the time of any death or human kill, the number of fertile animals, the number of pregnant females, the number of animals showing signs of toxicity, a description of the signs of toxicity observed, including time of onset, duration, and severity of any toxic effects, the types of histopathological changes, and all relevant litter data.

Reproductive indices:

Copulation index (%) = (numbers of animals mated/number of animals paired) × 100%
Fertility index (%) = (numbers of pregant female animals/number of animals paired) × 100%
Gestation index (%) = (numbers of pregnant female animals with live pups born/number of pregnant animals) × 100%
Viability index (%) = (numbers of pups alive Day 4 of age/numbe of pups born alive) × 100%

Clinical signs:

no effects observed

Dermal irritation (if dermal study):

not examined

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:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

effects observed, non-treatment-related

Description (incidence and severity):

No mortalities were observed in pregnant animals during the entire period of the study. All the parental animals mated successfully, and gave birth within 22 days of pregnancy. The number of animals with pregnancy period ≤21 days in LD group was significant reduced when compared to the control group (P<0.05), while no significant changes were noted in MD and HD groups (P>0.05). There were no significant changes observed in dams with live young born, dams with live young born at day 4 post-partum, number of corpora lutea/dam (mean) and number of implants/dam (mean) in treatment groups when compared to control group. No statistically significant difference was found in reproductive indices in treatment groups when compared to control group.

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

Clinical signs:

no effects observed

Description (incidence and severity):

No test item related clinical signs were observed in any pups during the 4 days post-partum.

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

For pups, 16 pups were born dead (R1014-F13 (control group), R2038-F19, R2042-F16 (low dose group), R3061-M7, R3064- M9, R3066-M6, R3071-M8, R3071-M9, R3071-M10, R3071-M11, R3071-F17 (intermediate dose group) and R4090-M5, R4090-F12, R4090-F13, R4090-F14, R4091-F15 (high dose group)) and 2 pups were dead at day 4 post-partum (R1018-F13 and R2044-M1). There were no significant differences in treatment groups were found in dead pups at birth when compared with control group.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

When compared with control group, no statistically significant differences were found in litter weight at birth (mean) (P>0.05) and litter weight at day 4 (mean) (P>0.05). There were significant reduction in pup weight at birth (mean), pup weight at day 4 (mean), in LD group when compared to control group, while no significant changes were found in MD and HD group. The significantly pup weight changes only noted in LD group was judged to be of no toxicological significance.

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Description (incidence and severity):

No abnormal pups observed (P>0.05).

Histopathological findings:

not examined

Other effects:

effects observed, non-treatment-related

Description (incidence and severity):

When compared with control group, no statistically significant differences were found in live/dead pups/dam at birth (P>0.05), live/dead pups/dam at day 4 (P>0.05), sex ratio (m/f) at birth (P>0.05), sex ratio (m/f) at day 4 (P>0.05), abnormal pups (P>0.05), loss of offspring (pre-implantation, pre-natal/post-implantations, post-natal). There were significant reduction in pup body length at birth (mean) and pup tail length at birth (mean) in LD group when compared to control group, while no significant changes were found in MD and HD group.

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

Key result

Dose descriptor:

NOAEL

Generation:

F1

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

Reproductive effects observed:

no

Results:

Table 1. Mean Relative Organ Weights to Body Weight

Group	Sex	n	Testes/ Uterus
			Left	Right
C	M	12	0.340±0.026	0.338±0.030
	F	9	0.232±0.042
LD	M	12	0.352±0.049	0.356±0.055
	F	11	0.276±0.043*
MD	M	12	0.340±0.039	0.354±0.034
	F	10	0.265±0.035
HD	M	12	0.375±0.039*	0.376±0.040*
	F	11	0.246±0.043

Data showed as Mean ± SD.

 

Table 2. Tabulated summary report of effects on reproduction/development

Observations		Values
Dosage (mg/kg)		0		100		300		1000
Females achieving pregnancy (N)		9		11		10		11
Conceiving days 1 - 5 (N)		9		11		10		11
Conceiving days 6 -14 (N)		0		0		0		0
Pregnancy ≤21 days (N)		5		11*		8		4
Pregnancy = 22 days (N)		4		0		2		7
Pregnancy≥23 days (N)		0		0		0		0
Pup weight at birth (mean±SD)		7.17±0.76		6.83±0.67*		7.19±0.62		7.09±0.70
Pup weight at day 4 (mean±SD)		9.76±1.42		8.86±1.26*		9.46±1.17		9.89±1.36
Pup body length at birth (mean±SD)		51.65±1.51		51.07±1.92*		51.76±1.17		51.88± 1.64
Pup tail length at birth (mean±SD)		16.96±1.53		16.56±1.23*		16.76±0.84		16.71± 1.28

* P<0.05, ** P<0.01

Conclusions:

In a reliable study conducted according to OECD 421 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 reproduction/developmental toxicity screening in males and females.

Executive summary:

Objective

The objective of this study was to assess the possible effects of the test item Aluminium Magnesium Vanadium Oxide on male, female fertility and embryofetal development in Sprague Dawley rats when administrated orally.

Study Design

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). The test item was administrated daily during 14 days pre mating period and 14 days mating period in both male and female, during gestation period and up to post natal day 4 in females. Animals were mated in the ratio of 1:1(Male to female). The non-pregnant females were still dosed to the day 24~26 post-mating.

During the period of administration, clinical signs, body weights, food consumption, fertility and mating performance, organ weights, and microscopic examinations were recorded for all parental animals. Litter size, litter observation, body weights, length and abnormalities were recorded for the offspring.

Results

No animal died and no test item related clinical signs were observed during the study. No statistically or toxicological significant effect was observed on body weight, food consumption, organ weights, histopathology findings, mating performanceand fertility, pregnancy and litter parameters in the study.

Conclusion

Based on the data generated from this study, the No Observed Adverse Effect Level (NOAEL) of Aluminium Magnesium Vanadium Oxide is 1000 mg/kg body weight/day for reproduction/developmental toxicity screening in males and females.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

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.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

A reproduction/developmental toxicity screening study was performed according to OECD Guidelines for Testing of Chemicals, 421,“Reproduction/Developmental Toxicity Screening Test”, adopted 27 Jul 1995. 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). The test item was administrated daily during 14 days pre-mating period and 14 days mating period in both male and female, during gestation period and up to post-natal day 4 in females. Animals were mated in the ratio of 1:1(Male to female). The non-pregnant females were still dosed to the day 24-26 post-mating.

There was no mortality among the P animals during the study period. No test item related clinical signs were observed in any male and female animals during the entire study period. No treatment-related effects were seen in the parental (P) animals on body weight and body weight gain, food consumption at any dose level.All the parental animals mated successfully, and gave birth within 22 days of pregnancy. The number of animals with pregnancy period≤21 days in LD group was significant reduced when compared to the control group (P<0.05), while no significant changes were noted in MD and HD groups (P>0.05). There were no significant changes observed in dams with live young born, dams with live young born at day 4 post-partum, number of corpora lutea/dam (mean) and number of implants/dam (mean) in treatment groups when compared to control group. The changes of relative organ weights of uterus were considered to be dose-related, however, the values of these changes remained within the range of the historical control data. There were no histopathological changes seen in the reproductive organs. Thus, the changes of relative organ weights were considered of no toxicological significance. The NOAEL for parental systemic toxicity was considered to be≥1000 mg/kg bw/day (males, females).

For pups, 16 pups were born dead and 2 pups were dead at day 4 post-partum. When compared with control group, no statistically significant differences were found in live/dead pups/dam at birth (P>0.05), live/dead pups/dam at day 4 (P>0.05), sex ratio (m/f) at birth (P>0.05), sex ratio (m/f) at day 4 (P>0.05), litter weight at birth (mean) (P>0.05), litter weight at day 4 (mean) (P>0.05), abnormal pups (P>0.05), loss of offspring (pre-implantation, pre-natal/post-implantations, post-natal). There were significant reduction in pup weight at birth (mean), pup weight at day 4 (mean), pup body length at birth (mean) and pup tail length at birth (mean) in LD group when compared to control group. The reason of these changes was supposed as the shorter gestation length in LD group, and these changes were not dose-related and considered to be of no toxicological significance. No test item related clinical signs were observed in any pups during the 4 days post-partum. A NOAEL for parental fertility of≥1000 mg/kg bw/day was derived for male and female rats.

Effects on developmental toxicity

Description of key information

Reproduction/Developmental Toxicity Screening Test (OECD 421), rat: 1000 mg/kg body weight/day (m/f)

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

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.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the available data, i.e. a Reproduction/Developmental Toxicity Screening Test and a 90-day repeated dose toxicity study, addressing toxicity to reproduction, aluminium magnesium vanadium oxide does not meet the criteria for classification for fertility and developmental toxicity according to Regulation (EC) No. 1272/2008. However, a final hazard conclusion and subsequent decision on classification for developmental toxicity cannnot be made as only a screening study according to OECD 421 is available. Therefore, classification for reproductive toxicity according to Regulation (EC) No. 1272/2008 remains open.