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Repeated dose toxicity: inhalation

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Administrative data

Endpoint:
short-term repeated dose toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Pre-GLP. No test material characterization information is available. Study was conducted in methods comparable to OECD guideline 412 "Repeated Dose Inhalation Toxicity: 28-day or 14-day Study". No histopathology was performed. The guideline indicates that 5 animals per sex per dose should be evaluated. In monkeys there were only two animals in each dose group. In guinea pigs there were six males and 6 females at each dose level, one-half of the animals in each group were sacrificed at the end of exposure, and the remaining animals were sacrificed after a 14-day recovery period. Thus, only 3 males and 3 females were analyzed after exposure or recovery period at each dose level. In rats the animal weight variation exceeded ± 20%. Although there were six males and 6 females at each dose level, one-half of the animals in each group were sacrificed at the end of exposure, and the remaining animals were sacrificed after a 14-day recovery period. Thus, only 3 males and 3 females were analyzed after exposure or recovery period at each dose level.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1972
Report date:
1971

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)
Deviations:
yes
Remarks:
See rationale for reliability above.
GLP compliance:
no
Remarks:
Study predates GLP
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Pentasodium triphosphate
EC Number:
231-838-7
EC Name:
Pentasodium triphosphate
Cas Number:
7758-29-4
Molecular formula:
H5O10P3.5Na
IUPAC Name:
pentasodium triphosphate
Details on test material:
- Name of test material (as cited in study report): XTW-449 (STP)
The study owner confirmed by email (8th Dec 2009) that according to their study records, the test substance XTW-449 was Sodium Tripolyphosphate (Hooker Chemical Co)

- Substance type: Inorganic
- Physical state: Solid

Test animals

Species:
other: Rat, Guinea pig and monkey
Strain:
other: Sprague-Dawley albino rats, Trueblood albino guinea pigs, Macaca irus (cynomolgus) monkey
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Sprague Dawley albino rats were obtained from Manor Research Staatsburg, New York, Trueblood guinea pigs Hazleton Research Animals, Burtonsville, Maryland. Sinomologus monkey Primate imports, New York and Woodard Research Virginia
- Age at study initiation: Not stated
- Weight at study initiation: Rate 200-300g. Guinea pig 300-320g
- Housing: Stainless steal wire mesh cages
- Diet (e.g. ad libitum): Once daily after daily exposure
- Water (e.g. ad libitum): Not stated


ENVIRONMENTAL CONDITIONS
No data

Administration / exposure

Route of administration:
inhalation: dust
Type of inhalation exposure:
whole body
Vehicle:
other: unchanged (no vehicle)
Remarks on MMAD:
MMAD / GSD: MMAD = 5.0 microns
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The chambers were fabricated from 304 L stainless steel with glass windows and are 6' x 6' x 6' with pyramidal tops and bottoms.
- Method of holding animals in test chamber: Each chamber also held four rodent cages in which the guinea pigs and rats were housed. The sexes of each species were separated from each other. The cages were made from stainless steel wire mesh to allow the maximum exposure of the rodents to airborne test substance. In some instances, the cages were placed in tiers and rotated on a daily basis In other cases.
- Source and rate of air: The test substance was introduced into a tube which is tangential to the mixing turrent on top of the chamber.
- Method of conditioning air: The test substance is carried along by room air which has passed through a charcoal bed and a Chambridge absolute filter.
- System of generating particulates/aerosols: Two methods for the generation of aerosols were employed in this experiment. When it was desirable to attain atmospheric concentrations of less than 20 micrograms per liter, the Wright Dust Feed was used. This type of dust generator operates by disseminating the powder through the operation of a series of different sized gears which makes it possible to accurately control the quantity of material metered into the chamber. To attain chamber concentrations from 20 ug/L to 500 ug/L, the pulse-puff aerosol generator was employed. This generator operates by the passage of compressed air through the bottom of the generator which suspends the powder in the generator. Blasts of air at regular intervals insures the suspension of the material which is then forced into the chamber plenum by an additional compressed air source at the top of the generator. The material was carried along into the mixing turret of the chamber by filtered house air which is continually being passed through the plenum.
- Temperature, humidity, pressure in air chamber: The temperature of these chambers was maintained at 75 F +/- 2 and the relative humidity was maintained at 50% +/- 5%.
- Air flow rate: The maximum airflow rate through the chamber was 1000 liters/minute.
- Air change rate: N/A
- Method of particle size determination: Atmospheric samples for particle sieve analysis were obtained with a Monsanto Cascade Impcator and an Andersen Particle Sizing Air Sampler.
- Treatment of exhaust air: Chamber was exhausted from the bottom of the chamber and passed through a particulate filter, an activated charcoal filter, and then through a final filter before being passed into the atmosphere.

TEST ATMOSPHERE
- Brief description of analytical method used: The atmospheric concentration of the test materials in the chambers was determined gravimetrically
- Samples taken from breathing zone: No

VEHICLE (if applicable)
N/A
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The sampled materials were collected by a MSA glass fiber filter which was connected to the sampling tube. These filters were weighed on an analytical balance. In each case the volume of air which was passed through the filter was recorded. The concentration of the aerosol in the chamber was determined by dividing the total quantity (weight) of powder collected by the volume (liters) of air in that sample. To ensure the validity of this method, these concentrations were corroborated by employing the Monsanto Cascade Impactor and the Anderson Particle-Sizing Air Sampler (Model II 0705) which are normally used for particle size determinations, but which can be employed also for the quantitation of atmospheric concentrations. In order to insure that the chamber concentrations remained relatively constant and at the proper concentration during the exposure, a number of samples were obtained during each exposure and immediately analyzed.
Duration of treatment / exposure:
6h per day
Frequency of treatment:
5d per week
Doses / concentrations
Remarks:
Doses / Concentrations:
12.6, 137.0 and 342.8 µg/L
Basis:
analytical conc.
No. of animals per sex per dose:
Rats 6. Guinea pigs 6. Monkey 2
Control animals:
yes, concurrent no treatment

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly


FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION: No data

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Rat Yes. Guinea pig No
- Time schedule for collection of blood: Blood was obtained from each rat at the time of sacrifice on Day 28 and Day 42. These blood samples were obtained from the abdominal aorta.
- Anesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: N/A
- Parameters checked: Hematocrit, Hemoglobin, Total Erythrocyte Counts, Total Leukocyte Counts, Differential Counts- Metamyelocytes, Immature Neutrophils, Neutrophils, Lymphocytes, Monocytes, Eosinophils, Basophils.

CLINICAL CHEMISTRY: Rat Yes. Guinea pig No
- Time schedule for collection of blood: Rats were measured on Day 28 and Day 42.
- Animals fasted: N/A
- How many animals: N/A
- Parameters checked: The biochemical determinations were performed on the serum automatically by a Technicon SMA-12 Auto Analyzer. Sodium, potassium, chloride, calcium, carbon dioxide, bilirubin, glucose, blood urea nitrogen, alkaline phosphatase, serum glutamic pyruvic transaminase, serum glutamic oxaloacetic transaminase, protein, albumin, globulin, albumin/globulin ratio, electrophoresis albumin, Alpha 1 fraction, Alpha 2 fraction, Beta fraction, Gamma fraction.

URINALYSIS: Rat Yes. Guinea pig No
- Time schedule for collection of urine: Overnight urine samples were obtained from all rats immediately prior to the initial inhalation exposure, and on Days 28 and 42.
- Metabolism cages used for collection of urine: Yes, the rats were placed in metabolism cages in the early afternoon, and the samples were collected the next morning.
- Animals fasted: Yes, the rats did not receive any food; however, water was available ad libitum.
- Parameters checked: calcium, copper, iron, magnesium, and zinc. The analysis was performed by atomic absorption.

NEUROBEHAVIOURAL EXAMINATION: No data

OTHER: Respiratory Physiology: Guinea pigs- Respiratory physiology was performed on all guinea pigs on Day 27, and on those remaining guinea pigs on Day 41. These pulmonary function tests included measurements of total respiratory flow resistance and the diffusing capacity of the lung.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes; All animals which died during the test period were immediately necropsied. One-half of all surviving animals were sacrificed and necropsied on Day 28; the remaining animals were necropsied on Day 42. At necropsy, the organs were observed for gross lesions.(see table 1)

HISTOPATHOLOGY: No. Organs where taken (see table 2) but no histopathological examination was performed
Statistics:
Chi square test to determine heterogeneity of means.
If there was significant heterogeneity among means, the student's t test was performed at the 0.05 probability level
When there was not significant heterogeneity among means, analysis of variance using the F-test was performed
The means of each experimental group were compared to the control group for the same interval.

Results and discussion

Results of examinations

Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
There were no pharmacotoxic signs exhibited by rats or guinea pigs exposed for four weeks to 12.6 ug/ L of the test substance. One guinea pig died in this group during the first week of the postexposure period.
Rats exposed to 137.0 ug/L of the test substance ostensibly were not adversely affected. Guinea pigs exposed to this concentration exhibited severe respiratory distress including dyspnea and gasping. Four of these animals died during the four-week exposure period. Dyspena was present during the entire four-week exposure period, but was not apparent during the two week postexposure period.
All animals exposed to 342.8 ug/L of the test material exhibited pharmacotoxic signs during the four weeks of exposure. Evidence of respiratory distress was apparent in both rats and guinea pigs. Eleven of twelve guinea pigs died by the third week of exposure.

BODY WEIGHT AND WEIGHT GAIN
The growth curves exhibited by male rats in the 12.6 ug/L (Group V) and 137.0 ug/L (Group VI) groups appeared normal throughout the four-week exposure period and two week post exposure observation period when compared to that of the control group (Group I). The growth curve of male rats in the 342.8 ug/L (Group VII) appeared to be slightly depressed. The growth curves exhibited by female rats in Groups V, VI, VII appeared normal throughout the four-week exposure period and two-week postexposure period. Weight loss incurred by all groups at Weeks 4 and 6 were the result of food deprivation of the rats while they were in metabolism cages for the collection of urine samples. The animal weight variation over the course of the study exceeded ±20%

The growth curve exhibited by Group V (12.6 ug/L) male guinea pigs was normal throughout the test period. That curve demonstrated by group VI (137.0 ug/L) also appeared normal although it represented only four guinea pigs at Weeks 2 and 3, and two animals at Week 4. The other animals in this group had died during the exposure period. A similar situation existed with Group VII (342.8 ug/L). Although the curve appeared relatively normal during the four-week exposure period, at Week 1 there were only two surviving animals, and only one survivor at Weeks 2, 3, 4, 5, and 6. The growth curve exhibited by female guinea pigs in Group VI (137.0 ug/L) appeared normal throughout the six-week test period. The curve demonstrated by Group V (12.6 ug/L) appeared somewhat depressed until Week 4, where an actual weight loss was incurred. There were no surviving guinea pigs in Group VII (342.8 ug/L) by Week 2 of the exposures.

FOOD CONSUMPTION
N/A

FOOD EFFICIENCY
N/A

WATER CONSUMPTION
N/A

OPHTHALMOSCOPIC EXAMINATION
N/A

HAEMATOLOGY
For purposes of statistical analysis, the male and female values for each parameter were combined and one mean was calculated. Any experimental mean which appeared abnormal was statistically compared to the control mean for that interval. The difference between the experimental group mean and the control group mean was significant at the 0.05 probability level as determined by the "t" test. Statistical analysis was performed on the following parameters: HCT (hematocrit), HGB (hemoglobin), RBC (red blood cells) and WBC (white blood cells). The differential counts were examined for abnormal values. The experimental hematological values were in the control or normal range for all groups at each interval, and there were no apparent trends of effect.

CLINICAL CHEMISTRY
Any experimental mean which appeared abnormal was statistically compared to the control mean for that interval. When the difference between the experimental group mean and the control group mean was significant at the 0.05 probability level as determined by the "t" test. Statistical analysis was performed on all parameters measured. There were several observations which may be of importance. The potassium (K+) level in the serum of the control rats was abnormally high at Day 28, however, the levels of the experimental groups were in the normal range. The means of the experimental groups listed were statistically different from the control mean at Day 28. The serum glutamic oxaloacetic transaminase (SGOT) increased for the rats receiving 137.0 ug/L. The ALB and A/G ratio increased in the rats which were in 12.6 ug/L, 137.0 ug/L and 342.8 ug/L groups. The globulin decreased in the 12.6 ug/L, 137.0 ug/L and 342.8 ug/L groups. The gamma-globulin decreased in the 12.6 and 137.0 ug/L group.

URINALYSIS

Rats:
Iron levels did not appear to be ostensibly increased or decreased at the Day 27 or Day 41 intervals when compared to the control group, Group I at those intervals, or each group's own pre-exposure control value. Zinc levels did not appear to noticeably increase or decrease at any interval when compared to the control group. Urine zinc levels were elevated above the pre-exposure values for Group V (12.6 ug/L) and VII (342.8 ug/L) at Day 27 but decreased to pre-exposure levels at Day 41. Calcium levels did not appear to be grossly increased or decreased at any interval when compared to the control group at that same interval. However, all levels, including the control level, increased substantially at the Day 27 and Day 41 intervals when compared to the pre-exposure values. At the Day 27 interval, the control value is elevated and is similar to its pre-exposure value. Groups V, VI, and VII (12.6, 137.0, and 342.8 ug/L of the test substance) also have elevated values at this interval which are almost identical to the control value. With the exception of Group VII, which were elevated, urinary magnesium levels of the test groups at Day 41 coincided with the control value.

NEUROBEHAVIOUR
N/A

ORGAN WEIGHTS
The organ/body weight ratios for rats and guinea pigs sacrificed at Days 28 and 42, and for guinea pigs which died at various times during the test period were recorded. A careful evaluation of the organ/ body weight ratios did not indicate seemingly abnormal values among any o f the animals which were sacrificed at the proper intervals, when compared to the control group. The organ/body weight ratios of guinea pigs which were found dead and then necropsied varied considerably since they were not exsanguinated and postmortem changes had often occurred.

GROSS PATHOLOGY
Rats:
No exposure-related gross lesions were seen in any tissues from rats immediately following four weeks of exposure to the test substance at 12.6, 137.0 and 342.8 ug/L or after the two-week postexposure observation period.

Guinea Pigs:
No consistent exposure-related gross lesions were observed in any of the tissues from Group V (12.6 ug/L) guinea pigs immediately following the four weeks of exposure or after the two-week observation period. Reddish and/or brownish discoloration of the lungs was noted in this group at Day 28 and 42. Reddish and/or brownish discoloration of the lungs was noted in those guinea pigs from group VI (137.0 ug/L) that died (six) during the study and in those that were sacrificed at the necropsy intervals. The lungs of animals which were not necropsed immediately upon death, i.e., those which were found dead in their cages, tend to be redder than normal since it was usually not possible to exsanguinate these animals. Eleven guinea pigs died in Group VII (342. 8 ug/ L) during the study. The lungs of these animals were discolored as mentioned previously. No other gross abnormalities were apparent.

HISTORICAL CONTROL DATA (if applicable)
N/A

OTHER FINDINGS

The weekly mean chamber concentrations +/- standard error as determined by gravimetric analysis were determined several times each day, five days a week, during the four-week exposure period. The arithmetic mean for four weeks of exposure for the test substance was 12.6, 137.0, and 342.8 ug/L. Particle size determinations were performed on all chambers during the four-week exposure period. Atmospheric samples for particle size analysis were obtained with a Monsanto Cascade Impactor and an Anderson Particle Sizing Air Sample. In every chamber, except the 150.0 ug/L chamber, the percent of particles less than 10 microns in diameter was greater than 80%. The mass median diameter of particles in all chambers was approximately 5.0 microns.

Respiratory Physiology:
Total Respiratory Flow Resistance
The results from the pulmonary function studies which were performed on guinea pigs are recorded. In all cases, the males and females in each group were combined in the calculation of group means. All respiratory physiology data were submitted to statistical analysis. Each experimental mean was compared to the control mean for that interval. When the difference between the experimental group mean and the control group mean was significant at the 0.05 probability level as determined by the "t" test. Total respiratory system flow resistance during expiration [Rrs(e)] was increased slightly in Group VI at Day 41. This value was not statistically significant when compared to the control valve for that interval. All other values were in the control or normal range.

Distribution of Inspired Air and Ventilation
There were not apparent increases in the number of breaths and times required to expire nitrogen from 80% to 1% in Groups V, VI, and VII. The respiratory rates from Groups V and VI were statistically elevated above the control group at the Day 27 interval. The mean tidal volumes of Group VI were increased above control values for those intervals, however, these increases were not statistically significant when compared to the controls.

Effect levels

open allclose all
Dose descriptor:
NOAEC
Effect level:
12.6 other: µg/L
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Dose descriptor:
NOAEL
Remarks:
Rat
Effect level:
> 342.8 other: µg/L
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The rat NOAEL was 342.8 ug/L. Statistically significant increases in serum GOT, albumin, globulin, total protein, , and gamma globulin were observed in rats exposed to test substance at Day 28. All parameters were normal after the 2-week rest period. No gross or histopathological effects were observed. The guinea pig NOAEL was 12.6 ug/L. Slight to severe respiratory distress was experienced at higher concentrations of test substance . Deaths occurred in guinea pigs exposed at 137 and 342.8 μg/L. There were some fluctuations in growth curves at the highest concentration. No gross or histopathological effects were observed.
Executive summary:

Rats and guinea pigs were exposed to graded airborne aerosol concentrations of the test substance for four weeks. The animals were exposed six hours a day, five days a week for four consecutive weeks. After the exposures were discontinued, one-half of the animals in each group were sacrificed and necropsies were performed. After a 14 day post exposure observation period, the remaining animals were sacrificed. All animals were observed daily for pharmacotoxic signs as manifestations of compound effect. All animals were weighed weekly during the six-week test period. Respiratory physiology, including measurements of total respiratory flow resistance, diffusion and ventilation was performed on all monkeys at pre-exposure, Day 27 and Day 41. Total respiratory resistance and diffusion were measured in all guinea pigs on Day 27 and Day 41. Blood was obtained from all rats at Days 28 and 42, and hematological and biochemical determinations were performed on the samples. Urine samples were obtained from all rats at pre-exposure, Day 28, and Day 42. These samples were subsequently analyzed for the trace metals: copper, iron, zinc, calcium, magnesium.

Respiratory distress, ranging from slight to severe, was experienced by guinea pigs exposed to the higher concentration of the test substance. Rats apparently were not affected at any of the levels employed. Deaths occurred in guinea pigs exposed to the test substance at 137.0 and 342.8 ug/L. Growth curves were not affected to any major extent in any of the groups of animals exposed to the various concentrations of the test materials. However, the guinea pigs did exhibit some fluctuations in growth curves at the highest concentration of each material.

Hematological determinations did not reveal consistent abnormal values in any of the experimental groups. Analyses of trace metals in the urine of rats did not reveal consistent abnormal values or trends of effect which could be directly attributed to exposures to any of the test materials. Pulmonary function studies on guinea pigs indicated various abnormalities which may or may not be importance when correlated to the results of histopathological examination of the lungs.

Gonadal tissues were examined for both gross pathology and histopathology and no treatment-related effects were detected.

Observations recorded at necropsy of rats and guinea pigs from the four week repeated exposure study, did not reveal consistent lesions which could be classified as compound related.

Although organs where taken for hystopathological examination, the study owner concluded no to perform this investigation and there is no histopathology report for the study.

The NOAEL for rats was determined to be >342.8 ug/L and the NOAEL for guinea pigs was determined to be 12.6 ug/L.