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Diss Factsheets

Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19.07.1976 to 17.07.1978 (for full study)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Cross-reference
Reason / purpose for cross-reference:
read-across: supporting information
Reference
Endpoint:
sub-chronic 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
Dose descriptor:
NOAEL
Effect level:
41 mg/kg bw/day (actual dose received)
Based on:
other: Effect level refers to disodium salt; equivalent to 34 mg/kg bw/day active acid
Sex:
male
Basis for effect level:
other: juvenile rats (500 ppm of HEDP (2-3Na)
Dose descriptor:
LOAEL
Effect level:
169 mg/kg bw/day (actual dose received)
Based on:
other: Effect level refers to disodium salt; equivalent to 139 mg/kg bw/day active acid
Sex:
male
Basis for effect level:
other: anaemia (2000 ppm HEDP (2-3Na)
Critical effects observed:
yes
Lowest effective dose / conc.:
169 mg/kg bw/day (actual dose received)
System:
haematopoietic
Organ:
blood
Treatment related:
yes
Relevant for humans:
not specified

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1977
Report date:
1977

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:
Conducted prior to guideline adoption.
GLP compliance:
no
Remarks:
pre-GLP
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium Salts of (1-Hydroxyethylidene)bisphosphonic acid (2-3 Na:1)
EC Number:
701-238-4
Molecular formula:
HEDP-2Na C2H6Na2O7P2 HEDP-3Na C2H5Na3O7P2
IUPAC Name:
Sodium Salts of (1-Hydroxyethylidene)bisphosphonic acid (2-3 Na:1)
Test material form:
not specified

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Anglia Laboratory animals, UK
- Age at study initiation: No data
- Weight at study initiation: 75-90 grams
- Fasting period before study: No
- Housing: Five per cage in suspended cages with wire-mesh floors.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: Six days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2 °C
- Humidity (%): 50 ± 5 %
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light


IN-LIFE DATES: From: 19.07.76 To: 17.07.78

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet: Weekly
- Mixing appropriate amounts with: Powdered laboratory rat food: Spratts Laboratory Diet 2.
- Storage temperature of food: No data
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Dietary samples were sent to the Sponsor for analysis of diets fed during week 30 and at approximately three month intervals thereafter. No further details provided. Therefore, dietary concentrations not verified during interim study period.
Duration of treatment / exposure:
90 days
Frequency of treatment:
continuous
Doses / concentrationsopen allclose all
Dose / conc.:
500 ppm
Remarks:
males: 41 mg/kg bw/day
females: 50 mg/kg bw/day (calculated by the reviewer)
Dose / conc.:
2 000 ppm
Remarks:
males: 169 mg/kg bw/day
females: 195 mg/kg bw/day (calculated by the reviewer)
Dose / conc.:
10 000 ppm
Remarks:
males: 817 mg/kg bw/day
females: 1000 mg/kg bw/day (calculated by the reviewer)
No. of animals per sex per dose:
Ten
Control animals:
yes, concurrent no treatment
Details on study design:
- Dose selection rationale: No data
- Rationale for animal assignment: Based on body weight
- Rationale for selecting satellite groups: Used to provide blood and urine samples during the first 26 weeks of the study, and therefore, subjected to the stresses of collecting these samples. Hence the main group animals were not subjected to these stressors until the end of the 102 week exposure period.
- Post-exposure recovery period in satellite groups: None
- Section schedule rationale: No data
Positive control:
None

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 for the first eight weeks, and two-weekly thereafter.

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: No, mean weekly intake calculated.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

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: Yes


WATER CONSUMPTION: Yes
- Time schedule for examinations: During week 4 for a 5-day period for each cage in control and high dose level main groups. During weeks 11 and 26 for a 5-day period for each cage of all main groups.

HAEMATOLOGY: Yes
- Time schedule for collection of blood and parameters measured: Weeks 0 and 5 from 10 males and females from Control and highest dose groups; Week 12 from 10 males and females in all groups; Weeks 25 from 10 males and females from control, mid and highest dose groups: packed cell volume, haemoglobin, red cell count, mean corpuscular haemoglobin concentration and mean cell volume, total white cell count and differential count. Platelet count and thrombotest were conducted in weeks 12 and 25 only. A visual estimation of red cell count and RBC osmotic fragility was conducted on the blood from high dose satellite group animals immediately prior to post-mortem.
- Anaesthetic used for blood collection: Yes (not identified)
- Animals fasted: Yes

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Week 5, 12 and 25: 5 males and 5 females from control and 10000 ppm; plasma urea, plasma glucose, total serum proteins, serum alkaline phophatase, serum glutamic pyruvic transaminase, sodium, potassium, calcium, inorganic phosphorus, serum creatinine. Week 7: 5 males from control, 2000 ppm and 10000 ppm for glucose and serum alkaline phosphatase. Week 12: 5 males from 500 ppm and 2000 ppm for serum alkaline phosphatase and serum glutamic pyruvic transaminase. Week 13: 5 females from all groups - plasma glucose. Week 25: 5 males from 2000 ppm group for serum alkaline phosphatase and 5 females from 2000 ppm group for plasma glucose.
- Animals fasted: Yes

URINALYSIS: Yes
- Time schedule for collection of urine: Weeks 6 and 25: 5 males and 5 females from control and highest dose: pH, specific gravity, protein, reducing substances, glucose, ketones, bile pigments, urobilinogen, haemoglobin, microscopy of spun deposits, urinary calcium, urinary phosphorus. Week 7: samples collected from 5 males and 5 females from all groups for estimation of pH, specific gravity and volume. During week 12: individual overnight urine samples from 5 males and 5 females from all groups. Urinary hydroxyproline measured in control and top dose at week 26.
- Metabolism cages used for collection of urine: No data
- Animals fasted: Yes

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see Table 1)
HISTOPATHOLOGY: Yes (see Table 1)
Other examinations:
None reported
Statistics:
One way analysis was performed on each parameter and treated groups compared with control using Student's t- test. Used for organ weight data, urinalysis, haematology, blood chemistry and bodyweights, food consumption, water consumption.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Severe pallor of skin in rats receiving 10 000 ppm and slight pallor in rats receiving 2000 ppm from week 6 were observed.
Mortality:
no mortality observed
Description (incidence):
Mortality occurred in one control female animal under anaesthesia for blood collection.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
During the first 12 weeks of treatment, a significantly reduced body weight gain was recorded for the 10 000 ppm group. There were no other statistically significant differences between the control and treated groups.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food intake was marginally (but statistically significant) reduced in highest dose group male rats. All other groups as for controls.
Food efficiency:
no effects observed
Description (incidence and severity):
No effect was observed in any of the dose groups.
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
In the highest dose groups, a reduced water consumption was observed which was in line with the reduced food consumption
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
During weeks 5 and 7, the 10000 ppm group had a statistically significant decrease in red cell parameters. Neutrophil and lymphocyte counts were significantly higher than controls. Additionally, a decrease in red cell values and higher neutrophil and lymphocyte counts for 2000 ppm males at week 7 (not studied at week 5) were observed. No similar statistically significant difference occurred for females. During week 12, a reduction in red cell parameters for both sexes at 10 000 ppm and for males at 2000 ppm was noticed. Moreover, a slightly higher platelet count for the 10000 ppm male group was observed. Examination of blood smears indicated a retardation of bone marrow development and prolonged anaemia at weeks 5, 7, 12 in both sexes at 10 000 ppm. During week 7, a slight retardation was seen in the 2000 ppm male group.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Higher alkaline phosphatase at week 5, 7 and 12 in males receiving 10 000 ppm was observed. No effects were observed in the 2000 ppm dose group at week 7. All other parameters were similar in control and treated males. Higher plasma glucose level in females receiving 10000 ppm at week 12 and 13 was observed, however, no effects in the lower dose females occurred.
Endocrine findings:
not examined
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Urinary volume was greater in the male treated groups at weeks 6 and 7, however, there were no differences at week 12. Non-dose-related sporadic increases in pH were detected in some rats, although this was not considered as of toxicological significance. Increased calcium in highest dose males at weeks 6 and 12 were reported, however, only when expressed in terms of volume of urine and not in absolute terms.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
There was a statistically significant decrease in liver weights recorded for males and females in the 10 000 ppm group. At 2000 ppm, the effect was observed in males, although at a significantly smaller magnitude. Additionally, a statistically significant decrease in kidney weight in the highest dose males was observed.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No findings of toxicological significance was observed.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no treatment-related findings.
Histopathological findings: neoplastic:
not examined
Other effects:
effects observed, treatment-related
Description (incidence and severity):
The bone marrow smears revealed a decrease in myeloid/erythroid and lymphocyte/erythroid ratios for rats in the highest dose group.

Effect levels

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
41 mg/kg bw/day (actual dose received)
Based on:
other: Effect level refers to disodium salt; equivalent to 34 mg/kg bw/day active acid
Sex:
male
Basis for effect level:
other: juvenile rats (500 ppm of HEDP (2-3Na)
Dose descriptor:
LOAEL
Effect level:
169 mg/kg bw/day (actual dose received)
Based on:
other: Effect level refers to disodium salt; equivalent to 139 mg/kg bw/day active acid
Sex:
male
Basis for effect level:
other: anaemia (2000 ppm HEDP (2-3Na)

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
In a well conducted, repeated oral dose toxicity study (reliability score 2), conducted using a protocol according to OECD 408 (Repeated Dose 90-Day Oral Toxicity in Rodents) and pre-GLP, the NOAEL for HEDP (2-3Na) was concluded to be 41 mg/kg bw/day (equivalent to 34 mg active acid/kg bw/day) based on anaemia at high doses.