Tris(pentane-2,4-dionato-O,O')vanadium

Toxicity to reproduction: other studies

Administrative data

Endpoint:

toxicity to reproduction: other studies

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1990/09/18 - 1990/12/20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

No ophthalmological examination

GLP compliance:

yes

Remarks:

The 3-month and 2-year studies were conducted in compliance with Food and Drug Administration Good Laboratory Practice Regulations (21 CFR, Part 58).

Type of method:

in vivo

Test material

Test material information

Constituent 1

Reference substance name:

Divanadium pentaoxide

EC Number:

215-239-8

EC Name:

Divanadium pentaoxide

Cas Number:

1314-62-1

Molecular formula:

O5V2

IUPAC Name:

dioxovanadiooxy(dioxo)vanadium

Test material form:

solid: particulate/powder

Details on test material:

Lot 1210490
EDX analyses of lot 1210490 indicated the presence of vanadium pentoxide with minor amounts of sulfur, chlorine, and potassium.

Specific details on test material used for the study:

Vanadium pentoxide was obtained from Shieldalloy Metallurgical Corporation (Newfield, NJ). Lot 1210490 was used in the 16-day and 3 months studies. Lot 1210490, an orange, crystalline solid, was identified as vanadium pentoxide by the analytical chemistry laboratory using X-ray diffraction (XRD) analyses and infrared and ultraviolet/visible spectroscopy and by the study laboratory using infrared spectroscopy. Infrared spectra were consistent between the lots used in different studies and with the structure of vanadium pentoxide (Nyquist and Kagel, 1971). XRD analyses of both lots indicated the presence of vanadium pentoxide with no detectable contaminants.

Test animals

Species:

other: Male and female F344/N rats and B6C3F1 mice

Strain:

other: Male and female F344/N rats and B6C3F1 mice

Administration / exposure

Route of administration:

inhalation: vapour

Type of inhalation exposure (if applicable):

whole body

Vehicle:

air

Details on exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Aerosol leaving the jetmill passed through a one-stage impactor and a vertical elutriator to eliminate or deagglomerate the large particles before entering a plenum and manifold distribution system. The aerosol delivery system consisted of three holding chambers that diluted the aerosol in three stages. A metered amount of diluted aerosol was removed and mixed with conditioned air at the inlet to each exposure chamber to achieve the appropriate exposure concentration. The electrical charge buildup on the aerosol particles was neutralized by mixing the aerosol with high concentrations of bipolar ions, which were generated using a Pulse Gun (Static Control Services, Palm Springs, CA) air nozzle. For the 3-month studies, a transvector air pump was installed at the aerosol inlet to each exposure chamber to provide additional control of the aerosol flow rate and improve stability of the chamber concentration.
- Method of holding animals in test chamber: not specified
- Source and rate of air: not specified
- Method of conditioning air: Chamber Air Supply Filters HEPA (R&R Equipment Sales, Rosemont, IL)
- System of generating particulates/aerosols: For the 16-day and 3-month studies, vanadium pentoxide aerosol generation was based on the principle of pneumatic dispersion and consisted of two major components: a screw feeder (Model 310, Accurate, White Water, WI) that metered vanadium pentoxide powder at a constant rate and a Jet-O-Mizer jetmill (Fluid Energy Corp., Harfield, PA) that used compressed air to disperse the metered powder and form the aerosol.
- Temperature, humidity, pressure in air chamber: 23.9°C, 55 +- 15 % humidity
- Air change rate: 15 changes/hour
- Method of particle size determination: The particle size distribution in each chamber was determined prior to the start of all studies, during the first week of the 16-day and 3-month studies, and monthly during the 3-month study. For the 16-day and 3-month studies, a 10-stage Quartz Crystal Microbalance-based cascade impactor (California Measurements, Inc., Sierra Madre, CA) was used to separate the aerosol particles into sequential size ranges; the mass median aerodynamic diameter was calculated from the corresponding mass fraction of particles at each stage.
- Treatment of exhaust air: not specified

TEST ATMOSPHERE
- Brief description of analytical method used: During all studies, chamber aerosol concentrations were monitored with real-time aerosol monitors (RAMs) that used a pulsed-light-emitting diode in combination with a silicon detector to sense light scattered over a forward angular range of 45° to 95° by particles traversing the sensing volume. The instruments respond to particles 0.1 to 20 μm in diameter.
- Samples taken from breathing zone: yes, an individual monitor was used for each exposure chamber

VEHICLE (if applicable)
- Justification for use and choice of vehicle: no details given
- Composition of vehicle: conditioned air
- Concentration of test material in vehicle: 0, 1, 2, 4, 8, or 16 mg/m3
- Purity of vehicle: not specified

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The uniformity of aerosol concentration in the inhalation exposure chambers without animals was evaluated before each of the studies began; concentration uniformity with animals present in the chambers was also measured once during the 3 months studies. RAM measurements were taken from 8 different chamber positions. During the 16-day and 3-month studies, minor excursions in chamber uniformity values (between-port and within-port variability) were observed in one or more exposure chambers, but these excursions had no impact on the studies.

Duration of treatment / exposure:

6 hours plus T90 (15 minutes) per day

Frequency of treatment:

5 days per week

Duration of test:

3 months

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/m³ air

Doses / concentrations 2

Dose / conc.:

2 mg/m³ air

Doses / concentrations 3

Dose / conc.:

4 mg/m³ air

Doses / concentrations 4

Dose / conc.:

8 mg/m³ air

Doses / concentrations 5

Dose / conc.:

16 mg/m³ air

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
rats: Based on decreased survival and body weight decreases in 32 mg/m3 in the 16 day study, an exposure concentration of 32 mg/m3 was considered too high for use in a 3-month study. Therefore, the exposure concentrations selected for the 3-month inhalation study in rats and mice were 0, 1, 2, 4, 8, and 16 mg/m3.
- Rationale for animal assignment (if not random): Animals were distributed randomly into groups of approximately equal initial mean body weights.

Statistics:

Survival Analyses
The probability of survival was estimated by the product-limit procedure of Kaplan and Meier (1958). Statistical analyses for possible doserelated effects on survival used Cox’s (1972) method for testing two groups for equality and Tarone’s (1975) life table test to identify dose-related trends. All reported P values for the survival analyses are two sided.

Analysis of Continuous Variables
Two approaches were employed to assess the significance of pairwise comparisons between exposed and control groups in the analysis of continuous variables. Organ and body weight data, which historically have approximately normal distributions, were analyzed with the parametric multiple comparison procedures of Dunnett (1955) and Williams (1971, 1972). Hematology, clinical chemistry, urinalysis, urine concentrating ability, cardiopulmonary, immunotoxicologic, cell proliferation, tissue concentrations, spermatid, and epididymal spermatozoal data, which have typically skewed distributions, were analyzed using the nonparametric multiple comparison methods of Shirley (1977) and Dunn (1964).

Results and discussion

Effect levels

Key result

Dose descriptor:

LOAEL

Remarks:

mice

Effect level:

8 mg/m³ air

Based on:

test mat.

Sex:

male

Basis for effect level:

other: epididymal sperm motility

Remarks on result:

other: The epididymal spermatozoal motility of males exposed to 8 or 16 mg/m3 was significantly decreased

Observed effects

The epididymal spermatozoal motility of male mice exposed to 8 or 16 mg/m3 was significantly decreased (Table 3). No significant differences were noted in estrous cycle parameters between exposed and chamber control female mice (Table 4). In rats no significant changes were observend in sperm motility. The estrous cycle length was significant longer than the chamber control in the 8 mg/m3 doese group, but not in the 16 mg/m3 dose group. (table 1 and 2).

Any other information on results incl. tables

Table 1 Summary of Reproductive Tissue Evaluations for Male Rats in the 3-Month Inhalation Study of Vanadium Pentoxide^a
	Chamber control	2 mg/m^3	4 mg/m^3	8 mg/m^3
n	10	10	10	10
Weights (g)
Necropsy body wt	350±8	339 ± 6	334 ± 7	319±7**
L. Cauda epididymis	0.1742±0.0062	0.1666±0.0042	0.1809±0.0043	0.1768±0.0056
L. Epididymis	0.4967±0.0076	0.4756 ± 0.0081	0.4896 ± 0.0097	0.5075 ± 0.0050
L. Testis	1.4960±0.0292	1.4606 ± 0.0326	1.5077 ± 0.0265	1.4908 ± 0.0183
Spermatid measurements
Spermatid heads (10^7/g testis)	9.70 ± 0.44	9.89 ± 0.58	9.35±0.39	8.91±0.42
Spermatid heads (10^7/testis)	14.42 ± 0.49	14.34 ± 0.72	14.08±0.59	13.30±0.69
Spermatid count (mean/10^-4mL suspension)	72.10±2.45	71.70±3.59	70.40 ± 2.94	66.50±3.45
Epididymalspermatozoal measurements
Motility(%)	85.68 ± 1.77	85.51 ± 2.21	87.69 ± 1.47	80.14±2.45
Concentration (10^6/g caudaepididymaltissue)	427 ± 15	463 ± 11	436±28	386±24
** Significantly different (P≤0.01) from the chamber control group by Williams' test ^aData are presented as mean ± standard error. Differences from the chamber control group are not significant by Dunnett's test (tissue weights) or Dunn's test (spermatid and epididymal spermatozoal measurements).

Table 2 Estrous Cycle Characterization for Female Rats in the 3-Month Inhalation Study of Vanadium Pentoxide^a
	Chamber Control	4 mg/m^3	8 mg/m^3	16 mg/m^3
n	10	10	10	8
Necropsy body wt	198 ± 3	196 ± 4	189 ± 4	117±5**
Estrous cycle length (days)	5.00 ± 0.00	5.00 ± 0.08	5.50±0.14**b	5.25± 0.25^c
Estrous stages (% of cycle)
Diestrus	39.2	40.8	49.2	71.9
Proestrus	18.3	16.7	15.8	10.4
Estrus	20.8	19.2	17.5	10.4
Metestrus	21.7	22.5	17.5	7.3
Uncertain diagnoses	0.0	0.8	0.0	0.0
** Significantly different (P≤0.01) from the chamber control group by Williams' test (necropsy body weight) or Shirley's test (estrous cycle length) ^aNecropsy body weight and estrous cycle length data are presented as mean ± standard error. By multivariate analysis of variance, exposed females do not differ significantly from the chamber control females in the relative length of time spent in the estrous stages. ^bEstrous cycle was longer than 12 days or was unclear in 1 of 10 animals. ^cEstrous cycle was longer than 12 days or was unclear in six of eight animals.

Table 3 Summary of Reproductive Tissue Evaluations for Male Mice in the 3-Month Inhalation Study of Vanadium Pentoxide^a
	Chamber Control	4 mg/m^3	8mg/m^3	16mg/m^3
n	10	10	10	9
Weights (g)
Necropsy body wt	35.4±1.1	34.5 ± 0.5	33.4±0.4	32.0±0.6**
L. Cauda epididymis	0.0170±0.0010	0.0174 ± 0.0006	0.0180±0.0006	0.0165±0.0009
L. Epididymis	0.0525±0.0012	0.0505 ± 0.0013	0.0546 ± 0.0013	0.0512±0.0013
L. Testis	0.1209±0.0025	0.1217±0.0014	0.1166±0.0020	0.1163±0.0018
Spermatid measurements
Spermatid heads (10^7/g testis)	17.97±0.67	15.99 ± 0.71	17.93±0.74	17.67±0.74
Spermatid heads (10^7/testis)	2.17±0.09	1.94 ± 0.08	2.09 ± 0.09	2.05 ± 0.08
Spermatid count (mean/10^-4mL suspension)	67.83±2.68	60.68 ±2.50	62.28 ± 2.89	64.06 ± 2.47
Epididymalspermatozoal measurements
Motility(%)	88.63± 0.90^b	86.23 ± 1.64	77.10± 3.15**^b	83.11± 2.48*^c
Concentration (10^6/g caudaepididymaltissue)	894 ± 57	915 ± 55	818 ± 39	849 ± 98
* Significantly different (P≤0.05) from the chamber control group by Shirley's test ** Significantly different (P≤0.01) from the chamber control group by Williams' test (necropsy body weight) or Shirley's test (epididymal spermatozoal motility) ^aData are presented as mean ± standard error. Differences from the chamber control group are not significant by Dunnett's test (tissueweights) or Dunn's test (spermatid measurements and epididymal spermatozoal concentration). ^bn=9 ^cn=8

Table 4 Estrous Cycle Characterization for Female Mice in the 3-Month Inhalation Study of Vanadium Pentoxide^a
	Chamber Control	4 mg/m^3	8 mg/m^3	16 mg/m^3
n	10	10	10	10
Necropsy body wt	31.1±1.0	26.2±0.4**	27.3 ± 0.4**	25.8 ± 0.4**
Estrous cycle length (days)	4.25 ± 0.13	4.29±0.15	4.05 ± 0.05	5.11± 0.51^c
Estrous stages (% of cycle)
Diestrus	27.5	40.8	29.2	34.2
Proestrus	21.7	14.2	15.0	18.3
Estrus	29.2	26.7	33.3	30.8
Metestrus	21.7	18.3	22.5	15.8
Uncertain diagnoses	0.0	0.0	0.0	0.8
** Significantly different (P≤0.01) from the chamber control group by Williams' test ^aNecropsy body weight and estrous cycle length data are presented as mean ± standard error. Differences from the chamber control group forestrous cycle length are not significant by Dunn's test. By multivariate analysis of variance, exposed females do not differ significantlyfrom the chamber control females in the relative length of time spent in the estrous stages. ^bEstrous cycle was longer than 12 days or was unclear in 3 of 10 animals. ^cEstrous cycle was longer than 12 days or was unclear in 1 of 10 animals.

Applicant's summary and conclusion

Conclusions:

The epididymal spermatozoal motility of male mice exposed to 8 or 16 mg/m3 was significantly decreased. No significant differences were noted in estrous cycle parameters between exposed and chamber control female mice. In male rats no significant changes were observend in sperm motility. The estrous cycle length in female rats was significant longer than the chamber control in the 8 mg/m3 doese group, but not in the 16 mg/m3 dose group.

Executive summary:

Groups of 10 male and 10 female rats were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 1, 2, 4, 8, or 16 mg/m3 by inhalation, 6 hours per day, 5 days per week for 3 months. Reproductive tissues were examined and estrous cycle and sperm measurements were performed. The following effects were observed: The epididymal spermatozoal motility of male mice exposed to 8 or 16 mg/m3 was significantly decreased. No significant differences were noted in estrous cycle parameters between exposed and chamber control female mice. In male rats no significant changes were observend in sperm motility. The estrous cycle length in female rats was significant longer than the chamber control in the 8 mg/m3 doese group, but not in the 16 mg/m3 dose group.