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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12.03.1985 to 17.04.1985 (inclusive of limit, range-finder and LD50 tests)
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Cross-reference
Reason / purpose for cross-reference:
read-across: supporting information
Reference
Endpoint:
acute toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
See read-across justification in IUCLID Section 13
Reason / purpose for cross-reference:
read-across source
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
2 850 mg/kg bw
95% CL:
>= 2 489 - <= 3 211
Remarks on result:
other:
Remarks:
(equivalent to 940 mg/kg bw active salt, which is equivalent to 659 mg/kg bw active acid)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1985
Report date:
1985

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
no post-dose fast
Principles of method if other than guideline:
Bio/dynamics protocol based on US guidelines. Broadly compatible with OECD guideline.
GLP compliance:
no
Remarks:
pre-GLP
Test type:
standard acute method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Tetrasodium (1-hydroxyethylidene)bisphosphonate
EC Number:
223-267-7
EC Name:
Tetrasodium (1-hydroxyethylidene)bisphosphonate
Cas Number:
3794-83-0
Molecular formula:
C2H8O7P2.4Na
IUPAC Name:
tetrasodium (1-hydroxyethane-1,1-diyl)bis(phosphonate)
Test material form:
liquid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: 9-12 weeks
- Weight at study initiation: 220-329 g
- Fasting period before study: 18 hours overnight prior to dosing
- Housing: Six per cage, suspended, stainless steel with wire mesh bottoms
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: At least 9 days


ENVIRONMENTAL CONDITIONS
- Temperature (°F): 67-76 (equivalent to 19.4-24.4 °C)
- Humidity (%): 30-70 %
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12 hours dark /12 hours light


IN-LIFE DATES: Limit test: From: 21.03.85 To: 22.03.85
Range-finding test: From: 27.03.85 To: 03.04.85
LD50 test: From: 03.04.85 To: 17.03.85

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 3.8 mL/kg in the limit test, 1.5 mL/kg in the range-finding test, and 3.1 mL/kg in the LD50 test.
Doses:
Limit test: 5000 mg/kg bw.
Range-finding test: 50, 100, 500, 1000 or 2000 mg/kg bw
LD50 determination: 2000, 2500, 3200 or 4000 mg/kg bw
No. of animals per sex per dose:
Limit test: 5
Range-finding test: 1
LD50 determination: 5
Control animals:
no
Details on study design:
Animals used in the range-finding test were observed for viability twice daily and deaths were recorded. The following observations were made on all other animals.
Viability check: twice daily.
Clinical signs: approximately 1, 2 and 4 hours after dosing and daily thereafter for 14 days
Body weights: Pre-fasting, post-fast, just prior to dosing, and Days 7 and 14 after dosing.
Macroscopic examination: Not on range-finding test animals. A macroscopic examination was conducted on all other animals.
Statistics:
The LD50 with 95% confidence limits was calculated using the method of Miller, Lloyd C. and M.L. Tainter., Estimation of the ED50 and its error by means of Logarithmic-Probit graph paper, Proc.Soc.Exp.Bio.Med. 57: 261-264 (1944).

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
2 850 mg/kg bw
95% CL:
>= 2 489 - <= 3 211
Remarks on result:
other: (equivalent to 940 mg/kg bw active salt, which is equivalent to 659 mg/kg bw active acid).
Mortality:
RANGE FINDING TEST: No deaths at any dose level. 
LD50 DETERMINATION TEST/LIMIT TEST: At 2000 mg/kg, 1 animal died (day 3 after dosing) 
At 2500 mg/kg, 4 animals died (1-4 hours after dosing) 
At 3200 mg/kg, 7 animals died (1 hour - day 8 after dosing) 
At 4000 mg/kg, 8 animals died (1-4 hours after dosing) 
At 5000 mg/kg, all 10 animals died (2-7 hours after dosing)
Clinical signs:
other: LD50 determination: Observations in all groups for up to four hours post-dosing, included ataxia and/or tremors, oral and nasal discharge, hypoactivity, soft stool and faecal and/or urinary staining.
Gross pathology:
LD50 determination: Macroscopic abnormalities observed at necropsy were primarily changes to the lungs (discolouration) and gastrointestinal tract (red or black walls, or red or black fluid present, suggestive of an irritant effect). Most of the animals in the 2500, 3200 and 4000 mg/kg dose groups had enlarged kidneys at necropsy, and one animal in the 4000 mg/kg bw group had unilateral renal pallor, dilated renal pelvis and red fluid surrounding the kidney. 
Other findings:
None reported.

Any other information on results incl. tables

Table 1 Summary of deaths in LD50 determination study.

 Dose   Mortality       Time to death
 mg/kg bw  Males  Females  Combined  
 2000  1/5  0/5  1  day 3
 2500  1/5  3/5  4  1 -4 h
 3200 3/5  4/5  7  1 h to day 8
 4000  4/5  4/5  8  1 -4 h
 5000  5/5  5/5  10  2 -7 h
 LD50  3100  2500  2850  
 95% conf limit  2365 -3635  1802 -3198  2489 -3211  




Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In an acute oral study, conducted using a protocol similar to the now deleted OECD Test Guideline 401 but pre-GLP, an LD50 of 2850 mg/kg was determined for HEDP-4Na in the rat. This is equivalent to 940 mg/kg/day bw active salt and 659 mg/kg bw/day active acid.