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Key value for chemical safety assessment

Effects on fertility

Description of key information
no data
Link to relevant study records
Reference
Endpoint:
toxicity to reproduction
Remarks:
other: Sub-chronic oral repeated dose toxicity study with histopathologic investigation of the reproductive organs.
Type of information:
experimental study
Adequacy of study:
weight of evidence
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.
Reason / purpose:
reference to same study
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:
no
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on mating procedure:
No mating
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
3 month
Frequency of treatment:
daily (continuous in the feed)
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
Remarks:
Doses / Concentrations:
200 ppm
Basis:
nominal in diet
approx. 15 mg/kg/day in males and 13 mg/kg/day in females
Remarks:
Doses / Concentrations:
1000 ppm
Basis:
nominal in diet
approx. 84 mg/kg/day in males and 125 mg/kg/day in females
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
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 (migrated information)
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 (migrated information)
Reproductive effects observed:
not specified

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

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.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
424 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The materials/methods and results are described sufficient for evaluation
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

There are no data available from a screening reproduction toxicity study or a two-generation reproduction toxicity study.

Tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate ("PBTCNa4") 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) (Löser E, Kaliner G, 1976).

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.

Taken together, the toxicological profile of PBTCNa4 and the corresponding parent acid 2-phosphonobutane-1,2,4-tricarboxylic acid ("PBTC") respectively is very consistent over a variety if different endpoints and parameters examined and the substance is shown to be of very low toxicity for any endpoint. Especially no effects are reported at any dose group in any parameter investigated, including gravimetric and histopathologic evaluation of reproductive organs, in a sub-chronic repeated dose toxicity study even at high doses 424 mg/kg/day (male rats) and 633 mg/kg/day (female rats), a dose very close to the limit dose for guideline studies. As outlined above repeated dose toxicity studies are considered to provide sufficient and sensitive information for fertility if histological examination of the reproductive organs is covered.


Short description of key information:
Taken together, based on the overall very low toxicity and the available data covering reproduction parameters, there is sufficient data available to conclude that tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate and the corresponding parent acid 2-phosphonobutane-1,2,4-tricarboxylic acid ("PBTC") respectively are not a reproduction toxicant.

Justification for selection of Effect on fertility via oral route:
Tetrasodium hydrogen 2-phosphonatobutane-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) (Löser E, Kaliner G, 1976). Toxicologically relevant information concerning reproduction can be deduced from this sub-chronic study because gravimetric and histopathologic investigation of the reproductive organs was reported.

Effects on developmental toxicity

Description of key information
Data on developmental toxicity are available in a prenatal developmental toxicity study similar to OECD TG 414 (Renhof, 1984).  Under the experimental conditions, the test item is considered to have no maternal or embryonic toxic effects and no teratogenicity effects in rats, even at the highest dose tested 1000 mg/kg/day (limit dose).
As there is no data available for tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate a read-across approach with the corresponding parent acid 2-phosphonobutane-1,2,4-tricarboxylic acid is proposed.
In aqueous media, tetrasodium hydrogen 2-phosphonatobutane-tricarboxylate and 2-phosphono-butane-1,2,4-tricarboxylic acid dissociate into the corresponding anion (2-phosphonatobutane-tricarboxylate ion) and the sodium ion and hydrogen ion (proton), respectively. The toxicological properties of 2-phosphonobutane-1,2,4-tricarboxylic acid and its tetrasodium salt are thought to be an effect of the phosphonato-carboxylate ion rather than of the sodium ion or the hydrogen ion (proton), which are normal constituents in body fluids and have no relevant toxic properties in low concentrations.
Therefore a read-across between tetrasodium hydrogen 2-phosphonatobutane-tricarboxylate and 2-phosphono-butane-1,2,4-tricarboxylic acid is justified.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1984-01 until 1984-02
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: scientifically aceptable and well documented
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
, the test substance was administered daily from day 6-15 of pregnancy (organogenesis period) instead of day 5-19 of pregnancy
Principles of method if other than guideline:
See above, deviations.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: sexually mature
- Weight at study initiation: Males-above 300 gr. Females-189-231 gr.
- Fasting period before study: Not indicated
- Housing: Makrolon cages type III (males), type II (females)
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: Not directly mentioned. At least 6 days before exposure to the test substance ( the test substance is given in the 6th day of pregnancy)


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22
- Humidity (%): 40%
- Air changes (per hr): Not mentioned
- Photoperiod (hrs dark / hrs light): 12 / 12 hours rhythm


Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
DIET PREPARATION
- Rate of preparation of diet (frequency): NA
- Mixing appropriate amounts with (Type of food): NA
- Storage temperature of food: NA


VEHICLE: Water
- Justification for use and choice of vehicle (if other than water): NA
- Concentration in vehicle: Test substance was dissolved in water and given in volume of 10 mL/kg to a final concentrations of 100 mg/kg bw, 300mg/kg bw, 1000 mg/kg bw
- Amount of vehicle (if gavage): See above
- Lot/batch no. (if required): Not required
- Purity: NA
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
NA
Details on mating procedure:
Cohoused:
- M/F ratio per cage: 1 male / 2 females
- Length of cohabitation: over night
- Proof of pregnancy: sperm in vaginal smear, referred to as day 0 of pregnancy
Duration of treatment / exposure:
The test substance was administrated from the 6th-15th day of pregnancy
Frequency of treatment:
Daily treatment
Duration of test:
20 days. At day 20 of pregnancy, embryos were taken out by abdominal delivery (cesarian surgery).
Remarks:
Doses / Concentrations:
0, 100, 300, 1000 mg/kg bw/day
Basis:
nominal in water
No. of animals per sex per dose:
25 Wistar rats per group (4 groups)
Control animals:
yes, concurrent no treatment
Details on study design:
- Dose selection rationale: NA
- Rationale for animal assignment (if not random): Random
Maternal examinations:
- Time schedule: Daily examination for changes in appearance and behaviour of the pregnant rats in all the four groups (control and other 3 different test substance concentration groups)
- Time schedule for examinations: During the whole pregnancy period
- Time schedule for examinations: Daily, by gavage.





Ovaries and uterine content:
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: No
- Number of implantations: Yes
- Number of early resorptions: No data
- Number of late resorptions: Yes
- Other: Weight of placenta
Fetal examinations:
- External examinations: Yes
- Soft tissue examinations: Yes
- Skeletal examinations: Yes
- Head examinations: No data
Statistics:
For the statistic calculations of the loss of weight, number of implantation and resorptions the WILCOXON-MANN-WHITNEY-U-TEST was used.
The Chiquadrat-Test was used for the statistic calculations of the embryotoxicity parameters
If not specified differently, the significant difference to control is under 5%.
Indices:
Examination for visceral modifications was according the modification of the WILSON technique.
The examination of the bones was done according to the DAWSON technique
Historical control data:
NA
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
NA - no effects
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
NA - no effects
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: teratogenicity
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: embryotoxicity
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: fetotoxicity
Abnormalities:
not specified
Developmental effects observed:
not specified

No remarks.

Conclusions:
No Maternal toxicity, no teratogenicity, no embryotoxicity under study conditions
Executive summary:

As there is no data available for tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate a read-across approach with the corresponding parent acid 2-phosphonobutane-1,2,4-tricarboxylic acid is proposed.

In aqueous media, tetrasodium hydrogen 2-phosphonatobutane-tricarboxylate and 2-phosphono-butane-1,2,4-tricarboxylic acid dissociate into the corresponding anion (2-phosphonatobutane-tricarboxylate ion) and the sodium ion and hydrogen ion (proton), respectively. The toxicological properties of 2-phosphonobutane-1,2,4-tricarboxylic acid and its tetrasodium salt are thought to be an effect of the phosphonato-carboxylate ion rather than of the sodium ion or the hydrogen ion (proton), which are normal constituents in body fluids and have no relevant toxic properties in low concentrations.

Therefore a read-across between tetrasodium hydrogen 2-phosphonatobutane-tricarboxylate and 2-phosphono-butane-1,2,4-tricarboxylic acid is justified.

After oral application of 2-phosphonobutane-1,2,4-tricarboxylic acid up to maximal dosage of 1000 mg/kg no signs of maternal toxicity were found (by means of death, weight loss, changes in appearance and behaviour). Moreover, female mother rats were proved later to be fertile. No influence was observed in embryo and foetus development (resorption, placenta weight, any skeletal and internal malformation). The NOEL value for these effects is therefore determined as 1000 mg/kg bw/day.

Under the experimental conditions, the test item is considered to have no maternal and embryonic toxic effects and no teratogenicity effects in rats.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Species:
rat
Quality of whole database:
The materials/methods and results are described in detail und are sufficient for evaluation
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Data on developmental toxicity are available in a prenatal developmental toxicity study similar to OECD TG 414 (Renhof M, 1984). The test compound, 2-phosphonobutane-1,2,4-tricarboxylic acid (PBTC), was administered by gavage to 25 female Wistar rats per group from gestation day 6 to 15 at dose levels of 0, 100, 300, and 1000 mg/kg bw/day. No signs of maternal toxicity were observed (incl. death, weight loss, changes in appearance and behaviour). No compound related effect was observed on fertility, embryo and foetus development (incl. resorptions, placenta weight, skeletal and internal malformation). Under the experimental conditions, the test item is considered to have no maternal or embryonic toxic effects and no teratogenicity effects in rats.

In aqueous media, PBTCNa4 and PBTC dissociate into the corresponding anion (2-phosphonatobutane-tricarboxylate ion) and the sodium ion and hydrogen ion (proton), respectively. The toxicological properties of PBTC and its tetrasodium salt are thought to be an effect of the phosphonato-carboxylate ion rather than of the sodium ion or the hydrogen ion (proton), which are normal constituents in body fluids and have no relevant toxic properties in low concentrations.

Therefore a read-across between PBTCNa4 and PBTC acid is justified.


Justification for selection of Effect on developmental toxicity: via oral route:
key study was used

Toxicity to reproduction: other studies

Additional information

no data

Justification for classification or non-classification

Based on the low toxicity profile and the available data on reproduction, there is sufficient data available to conclude that tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate and the corresponding parent acid 2-phosphonobutane-1,2,4-tricarboxylic acid respectively are not a reproduction toxicant.

Thus, according to EU Directive 67/548/EC and according to EU regulation No. 1272/2008 (GHS; amending and repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation (EC) No 1907/2006) tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate and the corresponding parent acid 2-phosphonobutane-1,2,4-tricarboxylic acid respectively should not be classified.