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Administrative data

Description of key information

Tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate was tested for acute oral (gavage) toxicity in 10 male rats by single applications that were well tolerated by all animals. The LD50 found was ca. 8300 mg/kg bw . This result is supported by an analogue study with tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate also in 10 rats. The LD50 (rat, oral) found was >1300 mg/kg based on active ingredient of a 32.6 % aqueous solution of tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate. Thus, tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate is not to be classified as acute toxic (oral route). 
Acute inhalation toxicity was determined for the test substance tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate. The LC50 (inhalation, rat) of tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate is >1979 mg/m3.
A dose level of 4000 mg/kg of the test substance tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate (32.6 % aqueous solution) was examined for acute dermal toxicity resulting in a LD50 (dermal, rat) of > 1300 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: short report
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Principles of method if other than guideline:
Ten male rats each received a single dose of 10 - 30 ml of the test substance per gavage. The animals were observed for mortality and clinical signs through day 14. A gross pathological examination was performed on animals which died during the observation period or were killed after termination of the study.
GLP compliance:
no
Test type:
standard acute method
Species:
rat
Strain:
Wistar
Sex:
male
Route of administration:
oral: gavage
Vehicle:
water
Doses:
10, 16, 18, 20, 22, 25, or 30 ml/kg bw of a 41.8 % solution in water
No. of animals per sex per dose:
10 animals/dose
Control animals:
no
Sex:
male
Dose descriptor:
LD50
Effect level:
ca. 8 300 mg/kg bw
Based on:
act. ingr.

sublethal range:

no findings

lethal range:

Beginning with an application volume of 16 ml/kg the animals showed after treatment a ruffle fur, decreased motility, prone position, accelerated and irregular breathing, cyanosis and after 24 hours diarrhoea. The animals died within 1 hour until 24 hours, but also up to 6 days after treatment. No cinical findings were evident 7 days after treatment in the surviving animals.

The section of the animals which died revealed a diffuse reddening of the mucosa of the stomach and intestine. The section of the surviving animals killed 14 days after treatment were without findings.

Interpretation of results:
relatively harmless
Remarks:
Migrated information
Executive summary:

Ten male rats each received a single dose of 10 - 30 ml of the test substance per gavage. The animals were observed for mortality and clinical signs through day 14. A gross pathological examination was performed on animals which died during the observation period or were killed after termination of the study. Signs of intoxication were ruffle fur, decreased motility, prone position, accelerated and irregular breathing, cyanosis and after 24 hours diarrhoea. The section of the animals which died revealed a diffuse reddening of the mucosa of the stomach and intestine. For male and female rats a LD50 = 20.1 ml/kg bw (ca. 8300 mg/kg bw) was found.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
8 300 mg/kg bw
Quality of whole database:
The materials/methods and results are described sufficient for evaluation

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Not indicated
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: scientifically acceptable and well documented
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
yes
Principles of method if other than guideline:
Observation period of 7 days (instead of 14 days).
GLP compliance:
not specified
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
- Wistar-Albino
- Weight at study initiation: 160-180 g.
- Diet: Standard lab diet, ad libitum
- Water: ad libitum
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
whole body
Vehicle:
other: water
Details on inhalation exposure:
- System of generating particulates/aerosols: Spray (aerosol) generated by dynamic inhalation apparatus.
- Brief description of analytical method used: A piece of cotton wool was applied with the sprayed test substance (as was inhaled by the rats). The sprayed cotton was dried in an oven (water elution) and weighed by a gravimetric method that gave the weight of the dried substance that was applied. The figure was divided to give a determination of mg test substance in cubic meters (m3) of air.
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Remarks on duration:
no
Concentrations:
800, 1479, 1979 mg/m3 air
No. of animals per sex per dose:
5 rats
Control animals:
no
Details on study design:
- Duration of observation period following administration: 7 days
- Other examinations performed: clinical signs, haematological parameters
Statistics:
NA
Preliminary study:
A concentration of 1780 mg/m3 air, was applied for one hour to 10 male and female rats. No clinical symptoms were observed.
Sex:
male/female
Dose descriptor:
LC0
Effect level:
1 979 mg/m³ air (analytical)
Exp. duration:
4 h
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 1 979 mg/m³ air (analytical)
Exp. duration:
4 h
Mortality:
No mortality during the time of the study.
Clinical signs:
No clinical symptoms appeared during the time of the study.
Body weight:
NA
Gross pathology:
NA
Other findings:
Haematological parameters: Erythrocytes, leucocytes, haemoglobin, haematocryte counts. No significant difference was found in the haematological results before and after the test substance application.

No remarks.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Inhalation of the test substance in concentrations of less than / equal 1979 mg/m3 is harmless. No mortality occurred and no clinical and haematological effects were observed.
Executive summary:

Acute inhalation toxicity was determined for the test substance, tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate. Five male and female rats were exposed for 4 hours to the test substance in analysed concentrations of 800, 1479, 1979 mg/m3 air. After 7 days of observations, no mortality occurred, no clinical symptoms were observed and no significant difference was found in haematological parameters which were examined before and after exposure.

The LC50 can be estimated as >1979 mg/m3. 1979 mg/m3 was the highest concentration that could technically be achieved. Following the mentioned results, the tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate is not to be classified as harmful via inhalation as at the highest achievable concentration (1979 mg/m3) no clinical symptoms were observed and no significant difference was found in haematological parameters which were examined before and after exposure.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating conc.
1 973 mg/m³
Quality of whole database:
The materials/methods and results are described in detail und are sufficient for evaluation

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1989-08-15 until 1989-08-29
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: scientifically acceptable and well documented
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Principles of method if other than guideline:
NA
GLP compliance:
yes
Test type:
standard acute method
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: Male rats: 9 weeks Female rats: 16 weeks
- Weight at study initiation: Male rats: 236 g Female rats: 217 g
- Housing:
- Diet: Conventional laboratory diet with ad libitum
- Water: Drinking water with libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/- 2°C
- Humidity (%): 50+/- 10%
- Air changes (per hr): 10 times
- Photoperiod (hrs dark / hrs light): 12 dark-/12 light

- In the end of the study (after 14 days) the rats, participated in the study were killed and subjected to pathological examinations.

Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: Back
- % coverage: Around 10%
- Type of wrap if used: Aluminium file
REMOVAL OF TEST SUBSTANCE
- Washing (if done): With warm water
- Time after start of exposure: 24 hours


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 4000 mg/kg
- Concentration (if solution): undiluted
- Constant volume or concentration used: no

Duration of exposure:
24 hours
Doses:
4000 mg/kg bw
No. of animals per sex per dose:
5 rats
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Few times at the day of application and twice a day in the following days until the end of the study.
Weighing was done before the application of the test substance, after one week and in the end of the 14 days study.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology.
Statistics:
NA
Preliminary study:
NA
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 1 300 mg/kg bw
Remarks on result:
other: based on active ingredient of a 32.6 % aqueous solution of tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate
Sex:
male/female
Dose descriptor:
LD0
Effect level:
1 300 mg/kg bw
Remarks on result:
other: based on active ingredient of a 32.6 % aqueous solution of tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate
Mortality:
No mortality during the time of the study.
Clinical signs:
Clinical signs were not observed during the study time.
Body weight:
In male rats the growing development was not disturbed. In female rats the weight changes were minor.
Gross pathology:
The pathological examination showed that one male rat had a brighter (paler) liver with stains. Another male rat had in addition to that a brighter (paler) kidney. Two of the female rats had also a brighter liver.
Other findings:
NA

No remarks.

Executive summary:

A 32.6 % aqueous solution (4000 mg/kg bw) of tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate was examined for acute dermal toxicity. 5 male rats and 5 female rats were observed for 14 days after dermal application of the test substance. No mortality occurred during the study time. The LD50 (rat, oral) was therefore estimated as > 1300 mg/kg based on active ingredient of a 32.6 % aqueous solution of tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate. No systemic clinical symptoms or local skin changes were observed. Minor changes were observed in body weight of female rats and no influence was observed in the growing development of the male rats. In a pathological examination of the study- rats at the end of the study, it was found that one male rat had a brighter (paler) liver with stains and another male rat had in addition a brighter kidney, two female rats, had also a brighter liver. Following the mentioned results, tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate is not to be classified as an acute toxic substance.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
1 300 mg/kg bw
Quality of whole database:
The materials/methods and results are described in detail und are sufficient for evaluation

Additional information

In acute toxicity testing, tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate ("PBTCNa4") revealed a low acute toxicity (oral, dermal and inhalation route):

Ten male rats each received a single dose of 10 - 30 ml of PBTCNa4 (41.8 % aqueous solution) per gavage. The animals were observed for mortality and clinical signs through day 14. A gross pathological examination was performed on animals which died during the observation period or were killed after termination of the study (Hoffmann, 1971).

Signs of intoxication were ruffle fur, decreased motility, prone position, accelerated and irregular breathing, cyanosis and after 24 hours diarrhoea. The section of the animals which died revealed a diffuse reddening of the mucosa of the stomach and intestine.

For male and female rats a LD50 = 20.1 ml/kg bw (ca. 8300 mg/kg bw) was found (Hoffmann, 1971).

PBTCNa4 (32.6 % aqueous solution) was tested for acute oral toxicity in 5 male and 5 female rats (Bomhard E, 1990). The test substance was applied one-time by gavage to a final amount of 4000 mg /kg bw.

After 14 days of observation no mortality occurred and no intoxication symptoms appeared. The LD50 (rat, oral) was therefore estimated as > 1300 mg/kg based on active ingredient of a 32.6 % aqueous solution of PBTCNa4.

The development of body weight was not effected in the male rats though a slight reduction in body weight of two male rats was documented. In the pathological examination of all study- rats at the end of the study, it was found that two male and 1 female rats exhibited a brighter (paler) kidney.

In conclusion, the LD50 (oral, rat) of PBTCNa4 is ca.8300 mg/kg bw according the study by Hoffmann, 1971.

Following the mentioned results, PBTCNa4 is not to be classified as acute toxic (oral route).

Acute inhalation toxicity was determined for the test substance, PBTCNa4 (41.4 % aqueous solution) as aerosol generated by a dynamic inhalation apparatus. Five male and female rats were exposed for 4 hours to the test substance in analysed concentrations of 800, 1479, 1979 mg/m³ air.

After 7 days of observations, no mortality occurred, no clinical symptoms were observed and no significant difference was found in haematological parameters which were examined before and after exposure. The LC50 can be estimated as >1979 mg/m³ (Mihail F, Kimmerle G, 1976).

In conclusion, the LC50 (inhalation, rat) of PBTCNa4, applied as aerosol is >1979 mg/m³ (>1.979 mg/L).

Following the mentioned results, PBTCNa4 is not to be classified as harmful via inhalation as at the highest achievable concentration (1979 mg/m³) no clinical symptoms were observed and no significant difference was found in haematological parameters which were examined before and after exposure.

A 32.6 % aqueous solution (4000 mg/kg bw) of PBTCNa4 was examined for acute dermal toxicity. 5 male rats and 5 female rats were observed for 14 days after dermal application of the test substance. No mortality occurred during the study time. The LD50 (rat, oral) was therefore estimated as > 1300 mg/kg based on active ingredient of a 32.6 % aqueous solution of PBTCNa4.

No systemic clinical symptoms or local skin changes were observed. Minor changes were observed in body weight of female rats and no influence was observed in the growing development of the male rats. In a pathological examination of the rats at the end of the study, it was found that one male rat had a brighter (paler) liver with stains and another male rat had in addition a brighter kidney, two female rats, had also a brighter liver (Bomhard E, 1990).

In conclusion, the LD50 (dermal, rat) of PBTCNa4 is >1300 mg/kg bw.

Depending on the high water solubility and the very low log Pow (equivalent to a very low lipophilic character), 2- PBTCNa4 are not likely to penetrate the first skin barrier (stratum corneum). Thus, upon dermal contact, the bioavailability of PBTCNa4 are expected to be low. Additionally PBTCNa4 has an oral LD50 of ca. 8300 mg/kg bw. As no mortality occurred at a dermal dose of 1300 mg/kg bw PBTCNa4, it is assumed, that the effective LD 50 is much higher than this value, but in any case higher than 2000 mg/kg bw.

Following the mentioned results, the PBTCNa4 is not to be classified as acute toxic (dermal route).

Based on the results of the acute toxicity studies, PBTCNa4 is not to be classified for acute toxic effects according to the CLP/ GHS.


Justification for selection of acute toxicity – oral endpoint
The most reliable study was used as key study and for classification

Justification for selection of acute toxicity – inhalation endpoint
only one study available

Justification for selection of acute toxicity – dermal endpoint
key study is used

Justification for classification or non-classification

Based on the results of acute toxicity testing tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate is not to be classified 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) as acute harmful (oral, dermal and inhalation route) due to its very low acute toxicity.