2-phosphonobutane-1,2,4-tricarboxylic acid

Toxicity to reproduction

Administrative data

Endpoint:

reproductive toxicity, other

Remarks:

other: Sub-chronic oral repeated dose toxicity study with histopathologic investigation of the reproductive organs.

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted 1976 using scientifically accepted methods similar to OECD TG 408.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1976

Report date:

1976

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: similar to OECD TG 408 (repeated dose 90 day oral toxicity in rats)

Deviations:

not applicable

Remarks:

no analytical verification of the test substance was reported

Principles of method if other than guideline:

Sub-chronic repeated dose toxicity study in rats including gravimetric and histopathologic investigation of the reproductive organs.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate

EC Number:

266-442-3

EC Name:

Tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate

Cas Number:

66669-53-2

Molecular formula:

C7H7Na4O9P

IUPAC Name:

hydrogen tetrasodium 2-phosphonatobutane-1,2,4-tricarboxylate

Details on test material:

technical sodium 2-phosphonatobutane-1,2,4-tricarboxylate

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- SPF
- Source: Winkelmann, Kirchborchen, Germany
- Age at study initiation: 28-32 days
- Weight at study initiation (male rats): Average 80-85 g
- Weight at study initiation (female rats): Average 78-81 g
- Housing: cages Makrolon, type II; 1 animals per cage
- Diet: ad libitum
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 +/-1°C

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on mating procedure:

No mating

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

NA

Duration of treatment / exposure:

3 month

Frequency of treatment:

daily (continuous in the feed)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

50 ppm

Remarks:

Doses / Concentrations:
50 ppm
Basis:
nominal in diet
approx. 4.2 mg/kg/day in males and 6.1 mg/kg/day in females

Doses / concentrations 2

Dose / conc.:

200 ppm

Remarks:

Doses / Concentrations:
200 ppm
Basis:
nominal in diet
approx. 15 mg/kg/day in males and 13 mg/kg/day in females

Doses / concentrations 3

Dose / conc.:

1 000 ppm

Remarks:

Doses / Concentrations:
1000 ppm
Basis:
nominal in diet
approx. 84 mg/kg/day in males and 125 mg/kg/day in females

Doses / concentrations 4

Dose / conc.:

5 000 ppm

Remarks:

Doses / Concentrations:
5000 ppm
Basis:
nominal in diet
approx. 424 mg/kg/day in males and 633 mg/kg/day in females

No. of animals per sex per dose:

test group: 15 animals / sex/ dose level
control group: 30 animals / sex

Control animals:

yes, plain diet

Details on study design:

Endpoints relevant for reproductive toxicity were included in this study as follows:



- Weights of reproductive organs (testes and ovaries) were investigated for all animals at the end of the experiment.



For histological examination of the following reproductive organs of five male and five female rats were reported:

- testes, prostate, seminal vesicle, ovaries and uterine horn ( highest dose group).

- testes and ovaries (all dose groups).



See cross reference to chapter 7.5.1. for a detailed description of all parameters investigated in this sub-cronic toxicity study similar to OECD TG 408.

CAGE SIDE OBSERVATION and DETAILED CLINICAL OBSERVATIONS: Yes
- The animals were observed for changes and symptoms daily (appearance, e.g. of fur, behaviour, e.g. concerning drinking and eating, motor activity ).

BODY WEIGHT: Yes
- The bodyweights of all involved animals were recorded weekly.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal was determined weekly. Based on that the mean daily consumption was calculated and given as g food/kg body weight/day: Yes
- 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: No data

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: One and three months after start of the feeding study
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 5 per sex and dose level
- Parameters examined: See "study design"

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: One and three months after start of the feeding study
- Animals fasted: No data
- How many animals: 5 per sex and dose level
- Parameters examined: See "study design

URINALYSIS: Yes
- Time schedule for collection of urine: Not indicated
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters examined: See "study design

NEUROBEHAVIOURAL EXAMINATION: No data

Results and discussion

Results: P0 (first parental generation)

Effect levels (P0)

1

Dose descriptor:

NOEL

Remarks:

systemic toxicity

Effect level:

5 000 ppm (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects at any parameter in the highest dose tested (ca. 424 mg/kg/day in males and 633 mg/kg/day in females)

Remarks on result:

other: Generation not specified.

2

Key result

Dose descriptor:

NOEL

Remarks:

reproductive organs

Effect level:

5 000 ppm (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect on weight or histopathology of reproductive organs up to the highest dose tested (ca. 424 mg/kg/day in males and 633 mg/kg/day in females)

Remarks on result:

other: Generation not specified.

Target system / organ toxicity (P0)

Critical effects observed:

no

Results: F1 generation

Effect levels (F1)

Remarks on result:

not measured/tested

Remarks:

sub-chronic oral repeated dose toxicity study with histopathologic investigation of the reproductive organs.

Target system / organ toxicity (F1)

Critical effects observed:

not specified

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Any other information on results incl. tables

Organ weights:

control/50/200/1000/5000ppm group

Males

Absolute testes weights:

3305/3415/3306/3328/3295 mg

Relative testes weight

972/1020/958/956/966 mg/100g body weight

Females

Absolute ovary weight:

92/94/97/97/105/89 mg

Relative ovary weight:

47/48/49/53/44

Histopathology:

dose 0 50 200 1000 5000

Seminal vesicles: no effect at any group

Prostate

No effect:        0/4 3/5

Epithelial proliferation                                    2/5

Testes/adrenals

Without findings                                   4/5 4/5 4/5 2/5 4/5

Cellular infiltration (very slight)                 1/5 1/5 1/5 3/5 1/5

Uterus/Ovaries

Without findings                                   3/5 0/5 1/5 2/5 2/5

Cellular dilation; endometrium                          1/5                                                  2/5

Dilation (unilateral)                                                             1/5                               1/5

Dilation (bilateral)                                             1/5             1/5              3/5

Cellular infiltration (slight)                                                    1/5             1/5                             2/5

Cellular infiltration (very slight)                                    2/5                                                1/5

Applicant's summary and conclusion

Conclusions:

Doses of up to 5000 ppm tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate applied over 3 months were tolerated without any effects.
The NOAEL of the tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate is equal or higher than 5000 ppm (equivalent to about 424 mg/kg bw for male rats and 633 mg/kg bw for female rats).

Executive summary:

The available information on repeated dose toxicity gave no evidence of any compound related effect in an early but comprehensive sub-chronic toxicity study similar to OECD TG 408 in which technical tetrasodium hydrogen 2-phosphonobutane-1,2,4-tricarboxylate was administered to 15 female and 15 male Wistar rats in diet, at dose levels of 0, 50, 200, 1000, and 5000 ppm (Löser E, Kaliner G, 1976).

 

 

All doses were tolerated without any compound related effect; for further details see chapter Repeated Dose Toxicity.

 

 

Consequently, it can be concluded that tetrasodium hydrogen 2-phosphonobutane-1,2,4-tricarboxylate is of very low systemic toxicity after sub-chronic exposure with NOEL levels at or above 424 mg/kg/day (males) and 633 mg/kg/day (females).

 

Toxicologically relevant information concerning reproduction can be deduced from this sub-chronic study because gravimetric and histopathologic investigation of the reproductive organs was reported. Testes and ovaries were weighted for all animals (15/sex/group). No compound related difference in organ weight was observed in any dose group. In addition histopathologic evaluation was performed in all dose groups (5/sex/dose). No compound related effect war observed in any reproductive organ investigated; seminal vesicle, prostate, testes and adrenals in males; uterus and ovaries in females.