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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
other: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
The study is conducted on a read across test material. The complete read across justification is attached in section 13. The reliability of the original study is 1.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BIOAGRI's animal house
- Weight at study initiation: Average of 244.8 g (males) 169.0 g (females)
- Age at study initiation: 7-8 weeks
- Housing: Polypropylene cages 3-2 rats/sex/group
- Diet and water ad libitum, regularly assayed for centesimal composition as well as for chemical and microbiological contaminant
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 hrs

Administration / exposure

Route of administration:
oral: feed
Vehicle:
other: Rodent diet
Details on oral exposure:
DIET PREPARATION WITH Test Item
- Nuvilab CR-1 diet type for rodents supplied by Nuvital nutrientes rtda (Brazil)
- Rate of preparation of diet (frequency): 10 days
- Mixing appropriate amounts with autoclaved feed through premix
- Storage temperature of food: room temperature

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentration and homogeneity of the test substance in the diet were determined by HPLC.
Difference between nominal and measured concentration was always within ±20 % acceptability
Duration of treatment / exposure:
Test duration: 28 days (4 weeks)
Satellite group was taken 2 more weeks without treatment.
Frequency of treatment:
Dosing regime: 7 days/week
Doses / concentrationsopen allclose all
Dose / conc.:
0 ppm
Remarks:
0 mg/kg bw/day
Dose / conc.:
100 ppm
Remarks:
equal to 9.9 mg/kg bw/day for males and 11.4 mg/kg bw/day for females
Dose / conc.:
300 ppm
Remarks:
equal to 27.8 mg/kg bw/day for males and 35.2 mg/kg bw/day for females
Dose / conc.:
1 000 ppm
Remarks:
equal to 86.9 mg/kg bw/day for males and 121.2 mg/kg bw/day for females
No. of animals per sex per dose:
Male: 5 animals at 0 ppm (0 mg/kg bw/day)
Male: 5 animals at 100 ppm (9.9 mg/kg bw/day)
Male: 5 animals at 300 ppm (27.8 mg/kg bw/day)
Male: 5 animals at 1000 ppm (86.9 mg/kg bw/day)
Female: 5 animals at 0 ppm (0 mg/kg bw/day)
Female: 5 animals at 100 ppm (11.4 mg/kg bw/day)
Female: 5 animals at 300 ppm (35.2 mg/kg bw/day)
Female: 5 animals at 1000 ppm (121.2 mg/kg bw/day)
Additional satellite groups (5 animals/sex/group) of the control and highest dose (1000 ppm) were maintained for more 2 weeks after the end of the treatment period for observation of reversibility or persistence of toxic effects
Control animals:
yes
Details on study design:
- Post-exposure recovery period in satellite groups: 2 weeks
Positive control:
None

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice a day for mortality and morbidity (once a day on Saturday and public holidays, no observation on Sunday)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- 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: Yes

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

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of the dosing regime
- Anaesthetic used for blood collection: Yes with CO2
- Animals fasted: Yes, overnight
- How many animals: all
- Parameters checked in table were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the dosing regime
- Anaesthetic used for blood collection: Yes with CO2
- Animals fasted: Yes, overnight
- How many animals: all
- Parameters checked in table were examined.

URINALYSIS: No

OTHER: Organ weight and histopathology
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Statistics:
Parametric: ANOVA followed by Dunnett's test
Non parametric: Wilcoxon, Kruskal Wallis
Fischer exact test was applied for macroscopic and histopathologic lesions
The level of significance was set at p < 0.05.
Statistical Program: SAS software v.8

Results and discussion

Results of examinations

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):
effects observed, non-treatment-related
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
Clinical observations:
- There was no mortality during the study.
- No significant clinical signs or disturbances of the general behaviour were detected in males and females of all tested groups.
- No alterations of mean body weight during treatment period.
- Statistical differences on food consumption observed in males during recovery period not related to treatment. No statistical differences on food consumption was observed on females during treatment or recovery periods.

Laboratory findings:
- Statistically significant increase on mean corpuscular hemoglobin concentration (MCHC) and platelet count (PLT) were observed on group 4 (1,000 ppm) of females, during treatment period. Statistically significant decrease on hematocrit (HCT) was observed on group 4 (1,000 ppm). No alteration on clotting and hematology parameters of males were observed in all tested groups during treatment period.
- No alterations on clinical chemistry of males were observed in all tested groups during treatment period.

Effects in organs:
- There were no alterations that could be attributed to the likely toxicity of the test item.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
ca. 35.2 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: 300 ppm, based on increased platelet count and mean corpuscolar haemoglobin concentration and a decrease of hematocrit. The results obtained for the satellite groups showed that these findings were fully reversible after recovery period.
Dose descriptor:
NOAEL
Effect level:
> 86.9 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: 1000 ppm. No substance related findings.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Repeated oral administration through diet of Phoslite IP-A to Wistar Hannover rats at doses of 0, 100, 300 and 1000 ppm for 28 days produced no mortality and no clinical signs of toxicological relevance.
Statistically significant increase on mean corpuscular hemoglobin concentration and platelet count were observed on group 4 (1,000 ppm) of females, during treatment period. Statistically significant  decrease on hematocrit (HCT) was observed on group 4 (1,000 ppm). 

Creatinine (CRE) concentration of females on group 4 (1,000 ppm) was statistically significant increased when compared to the control group.

Based on these results, the no-observed-adverse-effect-level (NOAEL) of Phoslite IP-A resulted to be higher than 300 ppm test item in diet for females (35.2 mg/kg body weight/day) and greater than 1000 ppm for males (86.9 mg/kg body weight/day).

Applicant's summary and conclusion

Conclusions:
NOAEL females = 35.2 mg/kg bw/day.
NOAEL males > 86.9 mg/kg bw/day.
There were no alteration that can be attributed to the toxicity of the test item. Minor alterations were detected in the female group at the highest dose (1000 ppm). All effects disappeared in the satellite group at the end of the recovery period.
Executive summary:

The test item was administered to Wistar rats (5 animals/sex/group) in the diet for 4 weeks at doses of 0, 100, 300, 1000 ppm, equal to compound intake of 0, 9.9, 27.8 and 86.9 mg/kg/bw on males and 0, 11.4, 35.2 and 121.2 mg/kg/bw on females. Additional satellite groups (5 animals/sex/group) of the control and highest dose were maintained for more 2 weeks after the end of the treatment period for observation of reversibility or persistence of toxic effects. Body weight and food consumption were determined weekly. Animals were checked daily for mortality and morbidity (except on Sundays) and were subjected to a careful clinical examination once a week. At the end of the treatment or after recovery period, blood samples were collected for clinical chemistry, hematological and clotting analyses. During the necropsy, the animals were subjected to a gross examination and the appropriate organs and tissues were removed, weighed and submitted for histopathological examination. In general, no substance related findings were observed. In the female group treated at the highest dose, increased on PLT and MHCH and decrease on HCT were recorded after treatment. The effects were fully reversible after the recovery period.