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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable publication which meets basic scientific principles; Data taken from EU risk assessment/SIDS

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Tributyl Phosphate Effects on Urine and Bladder Epithelium in Male Sprague-Dawley Rats
Author:
Arnold L.L. et al.
Year:
1997
Bibliographic source:
Fund. Appl. Toxicol. 40, 1997
Reference Type:
publication
Title:
OECD SIDS Ammonium Chloride, Final 8/2006
Author:
OECD
Year:
2006
Bibliographic source:
OECD SIDS AMMONIUM CHLORIDE

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
In this subchronic feeding study, ten male Sprague-Dawley received 12300 ppm ammonium chloride (equivalent to 684 mg/kg bw/day) continuously via diet for 70 days.
GLP compliance:
no
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Ammonium chloride
- Supplier: Sigma Chemical
- Analytical purity: > 98.7%

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories Inc. (Kingston NY)
- Age at study initiation: 4 - 5 weeks
- Weight at study initiation: mean 175 g
- Housing: individually in suspended stainless steel wire-mesh cages
- Diet (e.g. ad libitum): Purina Mills Certified Rodent Lab Chow, St. Louis, MO); ad libitum
- Water (e.g. ad libitum): water; ad libitum
- Acclimation period: 12 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/-4
- Humidity (%): 55 +/- 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The ammonium chloride content in the diet was determined by photometric analysis following extraction of chloride from feed using boiling water. The water/chloride extract was analyzed with a COBAS MIRA Plus Chemistry System (Roche Diagnostic System, Inc.)
Duration of treatment / exposure:
70 days
Frequency of treatment:
continously by feed
Doses / concentrations
Remarks:
Doses / Concentrations:
12300 ppm in the diet (equivalent to 684 mg/kg bw/day)
Basis:
nominal in diet
No. of animals per sex per dose:
10
Control animals:
yes, concurrent no treatment
Details on study design:
- Post-exposure recovery period: 7 days
Positive control:
Tributyl phosphate (TBP): 0, 3000 ppm (including 10 week recovery period)

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

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
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: No

URINALYSIS: Yes
- Time schedule for collection of urine: after each week
- Metabolism cages used for collection of urine: No
- Animals fasted: No data
- Parameters examined: pH, osmolality, Magnesium, Calcium, Creatinine, Phosphate, Protein

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes: bladder, kidney
HISTOPATHOLOGY: Yes: bladder, kidney

Results and discussion

Results of examinations

Details on results:
BODY WEIGHT
No significant difference between ammonium chloride treatment and control. Significantly decreased in the TBP treatment group.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
No significant difference among any of the groups (ammonium chloride, TBP and control); mean 110 g/kg b.w./day (initial) to 55.6 g/kg b.w./day (final)

URINALYSIS
- pH: ammonium chloride treatment reduced urinary pH to 6.04 (control: 7.56)
-Calcium: significantly increased in the ammonium chloride treatment. However the crystals from urine were not found.

ORGAN WEIGHTS
No significant difference compared to control


HISTOPATHOLOGY:
No abnormalities found.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
12 300 ppm
Sex:
male
Basis for effect level:
other: no adverse effects noted
Dose descriptor:
NOAEL
Effect level:
684 mg/kg bw/day (nominal)
Sex:
male
Basis for effect level:
other: no adverse effects noted

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Effects on urinary chemistry:

pH Magnesium (mEq/l) Osmolality (mOs/kg bw) Calcium (mg/dl) Creatinine (mg/dl) Phosphate (mg/dl) Protein (mg/dl)
NH4Cl 6.04 ± 0.10* 42.8 ± 11.3 1305 ± 156 11.0 ± 1.4* 52 ± 5+ 51 ± 9 108 ± 24
3000 ppm TBP 7.61 ± 0.05 37.6 ± 3.1 1034 ± 58* 4.5 ± 0.5 60 ± 4* 37 ± 11 119 ± 12
Control 7.56 ± 0.07 48.9 ± 5.1 1395 ± 87 5.9 ± 0.6 80 ± 5 41 ± 6 117 ± 11

* p<0.05

Histopathological effects on the bladder:

Bladder weight Bladder histology
g g/kg bw normal simple hyperplasia papillary/nodular hyperplasia BrdU labeling index
NH4Cl 0.138 ± 0.008 0.260 ± 0.014 10 0 0 0.17 ± 0.03
3000 ppm TBP 0.218 ± 0.027* 0.446 ± 0.052* 0 10* 6* 1.81 ± 0.30
Control 0.124 ± 0.010 0.227 ± 0.016 10 0 0 0.20 ± 0.03

* p<0.05

Applicant's summary and conclusion