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Environmental fate & pathways

Biodegradation in water: screening tests

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biodegradation in water: ready biodegradability
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
Not specified
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study, which is rated as reliability 2 because it is a read-across study.
Justification for type of information:
Please refer to IUCLID Section 13 for the read-across justification.
Reason / purpose:
read-across source
according to
OECD Guideline 301 E (Ready biodegradability: Modified OECD Screening Test)
GLP compliance:
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not specified
Oxygen conditions:
Inoculum or test system:
activated sludge (adaptation not specified)
Details on inoculum:
Return activated sludge from domestic wastewater treatment plant.
Duration of test (contact time):
28 d
Initial conc.:
100 mg/L
Based on:
test mat.
Details on study design:
Inoculum: The supernatant from the homogenized activated sludge was used as inoculum. The inoculum was pre-adapted to the test material for 14 days during which the test substance was added incrementally at concentrations equivalent to 4, 4 and 8 mg carbon/L on days 0, 7, and 12, respectively. The targeted microbial level in the test mixture was 10,000 to 1,000,000 cells/mL.
Concentration of test chemical: Test substance concentration was approximately 100 mg/L mineral medium, giving at least 50 to 100 mg ThOD per L medium. No organic solvents were used to facilitate the dispersion of the test material. The test substance was weighed onto a teflon coupon and introduced into the medium. Temp of incubation: 23 + 1ºC.
Dosing procedure: A measured volume of the inoculated mineral medium containing approximately 100 mg/L test substance is continuously stirred in a closed system for 28 days.
Sampling frequency: The oxygen uptake were monitored continuously and recorded every 4 hours throughout the test.
Controls: Yes (blank and positive controls per guideline); abiotic and toxicity checks were not included. Sodium benzoate was used as the positive control.
Method of calculating measured concentrations: N/A. Other: The inoculum was pre-adapted to the test substance for 14 days.
Reference substance:
benzoic acid, sodium salt
% degradation (TOC removal)
Sampling time:
28 d
Results with reference substance:
The reference substance, sodium benzoate, reached a level of 82.3% in the same test period and >60% in 3 days.
Validity criteria fulfilled:
not specified
Interpretation of results:
under test conditions no biodegradation observed
The test substance showed a low biodegradation rate (8.0%) in 28 days.
Executive summary:

Study conducted to OECD test guidelines in compliance with GLP. The test substance showed a low biodegradation rate (8.0%) in 28 days. The reference substance, sodium benzoate, reached a level of 82.3% in the same test period. Result: Under test conditions no biodegradation observed.

Description of key information

Based on read-across, the substance is considered to be not readily biodegradable.

Key value for chemical safety assessment

Biodegradation in water:
under test conditions no biodegradation observed

Additional information

Read across to supporting substance, CAS 36878 -20 -3, by structural analogue. Structure attached below. This category of substances is not presumed to biodegrade.

This substance has been supported under Environmental Protection Agency’s (EPA’s) High Production Volume (HPV) Challenge Program. The American Chemical Councils RAPA Panel, has derived a “Substituted Diphenylamines” category of chemicals for this substance, please refer to EPA reference 201-14700A located at


Relying on several factors specified in EPA’s guidance document on “Development of Chemical Categories in the HPV Challenge Program,” in which use of chemical categories is encouraged, the chemicals constitute a chemical category on the following basis:


Structural Similarity.A key factor supporting the classification of these chemicals as a category is their structural similarity (see Figure 1). All share a common starting material; Diphenylamine (Benzenamine, N-phenyl-, CAS# 122-39-4), a common synthetic pathway, and all compounds in this category are diamines with various substitutions.


Similarity of Physicochemical Properties.The similarity of the physicochemical properties of these materials parallels their structural similarity. All are off-white to light brown solids or viscous liquids intended for use as antioxidants in finished rubber articles or as antidegradant additives that extend the useful life of heavy-duty industrial functional fluids used in high-speed, high-temperature and/or high-load applications. As a class, these amine-based antidegradant compounds are less migratory (more polymer-bound) and less staining than the Substituted p-Phenylenediamine antidegradants. The use of these materials requires that they be stable under high temperatures. Their low volatility is due to their low vapor pressure, highly viscous or solid form. The existing information for these materials indicates that they have low water solubility and high flash points.


Fate and Transport Characteristics.Members of this category have been shown to be not readily biodegradable, so additional testing is not needed. The lack of water solubility of the members of this category makes hydrolysis testing unnecessary. These materials have been shown not to partition to water or air if released into the environment due to their low water solubility and low vapor pressure.


Conclusion.Based upon the data reviewed in “Substituted Diphenylamines” category of chemicals, the physicochemical and toxicological properties of the Substituted Diphenylamine category members are similar and follow a regular pattern as a result of that structural similarity. Therefore, the definition of a chemical category has been met, and read across is considered appropriate for the category of chemical.  This category of substances is not known to be readily biodegradable under standard conditions.