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EC number: 266-442-3 | CAS number: 66669-53-2
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
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- Boiling point
- Density
- Particle size distribution (Granulometry)
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- Endpoint summary
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- Environmental data
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- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
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- Toxicological Summary
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- Additional toxicological data

Toxicity to microorganisms
Administrative data
Link to relevant study record(s)
- Endpoint:
- activated sludge respiration inhibition testing
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 11/2008
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- no
- Vehicle:
- no
- Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- Origin: aeration tank of a domestic sewage treatment plant (Municipal STP Cologne-Stammheim)
Laboratory culture:Date of collection: 2008-11-24
Microbial inoculum:
- first, the sludge was settled and the supernatant was decanted
- after centrifuging (15 min at 4000 rpm and 20°C) the supernatant was decanted
- approx. 1 g of the wet sludge was dried in order to calculate the amount of wet sludge to achieve a concentration of activated sludge of 3 g/L (dry weight) suspended solids
- the calculated amount of sludge was first dissolved in synthetic medium and than filled up to a defined end volume with deionised water - before start of the test 10 mL of the sludge suspension were sucked of and the dry weight was controlled again. The sludge suspension was diluted with water to obtain a concentration of activated sludge of 2 g/L (dry weight) suspended solids
Storage of sludge: aeration of the activated sludge at 20°C ± 2°C, daily fed with synthetic medium
pH of the suspension before application: 7.7 - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Test temperature:
- Test item: 18.5 - 18.7 °C
- pH:
- Test item: 8.0 - 8.1
- Dissolved oxygen:
- Start: 6.2 - 6.4 mg O2/L
End: 2.8 - 3.7 mg/L - Nominal and measured concentrations:
- Nominal: 10; 100; 1000 mg/L
- Reference substance (positive control):
- yes
- Remarks:
- 3.5-Dichlorophenol
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 504 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks:
- respiration rate
- Remarks on result:
- other: The 50.4% aqueous solution of 2-phosphonatobutane-1,2,4-tricarboxylate acid showed 4.3 % respiration inhibition of activated sludge at the highest test item concentration of 1000 mg/L corresponding to a concentration of 504 mg/L of PBTC
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- > 504 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks:
- respiration rate
- Remarks on result:
- other: The 50.4% aqueous solution of 2-phosphonatobutane-1,2,4-tricarboxylate acid showed 4.3 % respiration inhibition of activated sludge at the highest test item concentration of 1000 mg/L corresponding to a concentration of 504 mg/L of PBTC
- Results with reference substance (positive control):
- The results with 3.5-Dichlorophenol:
15% inhibition and 18.0 mg/L*h at a concentration of 5 mg/L.
59.1% inhibition and 8.7 mg/L*h at a concentration of 10 mg/L.
83.8% inhibition and 3.4 mg/L*h at a concentration of 20 mg/L. - Reported statistics and error estimates:
- probit analysis
- Validity criteria fulfilled:
- yes
- Conclusions:
- The substance showed 4.3 % respiration inhibition of activated sludge at a test item concentration of 1000 mg/L. The 3h-EC50 is higher than 1000 mg/L. Based on the PBTC concentration in the test item (50.4%), the result can be converted yielding a 3h-EC50 of 504 mg/l for the further consideration of this study.
- Executive summary:
A study was performed to assess the toxicity of 2-phosphonatobutane-1,2,4-tricarboxylate acid to microorganisms. The study was conducted in accordance with Council Regulation (EC) No 440/2008, Method C.11 "Activated sludge respiration inhibition" (2008). This test method is equal to OECD Guideline 209 (1984). The activated sludge was exposed to a 50.4% aqueous solution of 2-phosphonatobutane-1,2,4-tricarboxylate acid at different concentrations. The respiration rate of each mixture was determined after aeration periods of 3 hours. The 50.4% aqueous solution of 2-phosphonatobutane-1,2,4-tricarboxylate acid (marketed as Bayhibit AM) showed 4.3 % respiration inhibition of activated sludge at a test item concentration of 1000 mg/L. The 3h-EC50 and the 3h-EC10 are therefore higher than 1000 mg/L. The effect value related to nominal concentration, since no analytical monitoring was performed. Based on the PBTC concentration in the test item (50.4%), the result can be converted yielding a 3h-EC50 of 504 mg/l for the further consideration of this study.
Reference
Table: Results of test item
Test item concentration (nominal) [mg/L] |
Respiratory rate test item [mg/L*h] |
Phys.-chem. O2consumption [mg/L*h] |
Respiratory rate - phys.-chem. O2consumption [mg/L*h] |
Inhibition [%] |
10 |
23.3 |
0.0* |
23.3 |
0.0 |
100 |
22.7 |
0.0* |
22.7 |
0.0 |
1000 |
20.3 |
0.0 |
20.3 |
4.3 |
* The physico-chemical oxygen consumption has been determined at 1000 mg/L test item concentration. As no physico-chemical oxygen consumption was observed at that test item concentration this observation also holds true for the lower test item concentrations.
Description of key information
As there is no data available for tetrasodium hydrogen 2-phosphonatobutane-1,2,4-tricarboxylate, results of the parent acid, 2-phosphonobutane-1,2,4-tricarboxylic acid are taken into account for this endpoint.
A 50.4% aqueous solution of 2-phosphonobutane-1,2,4-tricarboxylic acid showed 4.3 % respiration inhibition of activated sludge at a test item concentration of 1000 mg/L. The EC50 is higher than 1000 mg/L. Based on the amount of substance in the test item, the result can be converted yielding an EC50 of 504 mg/l for the further consideration of this study.
Key value for chemical safety assessment
- EC50 for microorganisms:
- 504 mg/L
- EC10 or NOEC for microorganisms:
- 504
Additional information
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. Fate, behavior and the ecotoxicological 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 environmental systems and have no relevant ecotoxic properties in low concentrations.
Therefore a read-across between tetrasodium hydrogen 2-phosphonatobutane-tricarboxylate and 2-phosphonobutane-1,2,4-tricarboxylic acid is justified.
To assess the toxicity of 2-phosphonobutane-1,2,4-tricarboxylic acid to microorganisms, a study was conducted in accordance with Council Regulation (EC) No 440/2008, Method C.11 "Activated sludge respiration inhibition" (2008) - equivalent to OECD Guideline 209 (1984). The activated sludge was exposed to a 50.4% water solution of 2-phosphonobutane-1,2,4-tricarboxylic acid at different concentrations. The respiration rate of each mixture was determined after aeration periods of 3 hours. 2-phosphonobutane-1,2,4-tricarboxylic acid solution showed 4.3 % respiration inhibition of activated sludge at a test item concentration of 1000 mg/L. The EC50 is higher than 1000 mg/L. The effect value related to nominal concentration of the water solution, since no analytical monitoring was performed. Therefore the effect concentration can be converted based upon the water content of the test item, resulting in an EC10 and EC50 of 504 mg/l for further consideration of this study.
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