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EC number: 807-130-4 | CAS number: 53716-82-8
- Life Cycle description
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- Ecotoxicological Summary
- Aquatic toxicity
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- Short-term toxicity to fish
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Biodegradation in water: screening tests
Administrative data
Link to relevant study record(s)
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 07 Apr - 07 May 2014
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 A (Ready Biodegradability: DOC Die Away Test)
- Version / remarks:
- 1992
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.4-A (Determination of the "Ready" Biodegradability - Dissolved Organic Carbon (DOC) Die-Away Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 835.3110 (Ready Biodegradability)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Swiss Federal Office of Public Health, Consumer protection directorate, Notification authority for chemicals, Bern, Switzerland
- Specific details on test material used for the study:
- Reference Item:
The reference item and the following information were supplied by Sigma-Aldrich:
Identity: Sodium benzoate
Product No.: 71295
Lot No.: BCBB4145
Purity: 99.7%
Expiration Date (as given by the supplier): November 2015 (date of recommended retest)
Expiration Date (handled at Harlan Laboratories): 30 November 2015 (date of recommended retest)
Storage Conditions (as provided by the supplier): Keep container tightly closed in a dry and wellventilated place. Store in cool place.
Storage Conditions (as handled at Harlan Laboratories): In tightly closed original container, at room temperature at about 20°C. - Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- - Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): wastewater treatment plant (ARA Ergolz II, Füllinsdorf / Switzerland)- Preparation of inoculum for exposure: The sludge was washed twice with tap water by centrifugation and the supernatant liquid phase was decanted. A homogenized aliquot of the final sludge suspension was weighed, thereafter dried and the ratio of wet to dry weight was calculated. Based on this ratio, calculated amounts of wet sludge were suspended in mineral medium to obtain a concentration equivalent to 4 g dry material per liter. During the holding period of one day prior to use, the sludge was aerated at room temperature. Prior to use, the dry weight of this diluted activated sludge was determined again and defined amounts were added to mineral medium to obtain a final concentration of 30 mg dry material per liter.
- Duration of test (contact time):
- 28 d
- Initial conc.:
- ca. 50 - ca. 50.7 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- DOC removal
- Details on study design:
- TEST CONDITIONS-
Composition of medium:
The mineral medium was prepared according to the testing guidelines. Analytical grade salts were dissolved in purified water to obtain the following stock solutions:
1) KH2PO4: 8.50 g/L, K2HPO4: 21.75 g/L, Na2HPO4 × 2H2O: 33.40 g/L, NH4Cl: 0.50 g/L. The pH of this solution was 7.4.
2) MgSO4 × 7H2O: 22.50 g/L.
3) CaCl2 × 2H2O: 36.40 g/L.
4) FeCl3 × 6H2O: 0.25 g/L, stabilized with one drop of concentrated HCl per liter.
To obtain the final mineral medium, 10 mL of stock solution 1) and 1 mL each of stock solutions 2) to 4) were combined and made up to 1000 mL with purified water. The pH was adjusted from 8.1 to 7.4 with a diluted hydrochloric acid solution.
TEST SYSTEM- Culturing apparatus: 2000-mL Erlenmeyer flasks, cleaned with alcoholic hydrochloric acid, rinsed with deionized water and dried. The final volume was 1000 mL per flask. Each flask was loosely covered with aluminium foil to allow the exchange of air between the flask and the surrounding atmosphere. The test media were continuously stirred by magnetic stirrers.
- Number of culture flasks/concentration: 2
- Test performed in open system: loosely covered with aluminium foil
- Test temperature: 21-24°C. The inoculated flasks were incubated in a temperature controlled room. The temperature was checked on each sampling date in one of the inoculum controls. Additionally, the room temperature was continuously recorded.
- pH: Prior to test start, the pH was measured in each test flask before the addition of the activated sludge inoculum and found to be pH 7.4. At the end of incubation, the pH was measured again in each test flask.
- Light conditions: The test flasks were incubated in a dark room.
SAMPLING- Sampling frequency: All vessels were sampled at Exposure Day 0, 2, 5, 8, 11, 14, 21 and 28.
- Sampling method: One sample of about 10 mL was taken from each test flask per sampling date. Prior to sampling, water evaporation losses were determined by weighing the flasks and were compensated by adding purified water. Deposits on the test vessels were resuspended.
CONTROL AND BLANK SYSTEM- Inoculum blank: yes (2)
- Toxicity control: yes (1)- Procedure control: yes (2) - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (DOC removal)
- Value:
- 25
- Sampling time:
- 11 d
- Key result
- Parameter:
- % degradation (DOC removal)
- Value:
- 99
- Sampling time:
- 14 d
- Key result
- Parameter:
- % degradation (DOC removal)
- Value:
- 99
- Sampling time:
- 28 d
- Results with reference substance:
- In the procedure controls, containing the reference item sodium benzoate and activated sludge (inoculum), sodium benzoate was readily biodegraded by an average of 98% within two days of exposure, thus confirming suitability of the activated sludge.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- readily biodegradable
- Conclusions:
- A biodegradation of 99% in 28 days (DOC removal) was determined for the substance using a relevant test method and in compliance with GLP. The result is considered to be reliable.
Reference
Degradation of the Test Item:
In the test flasks, containing the test item Cyrene™ and activated sludge (inoculum), the initial mean concentration of dissolved organic carbon (DOC) of 28.7 mg/L on Day 0 rapidly decreased after Exposure Day 8, reaching 99% decrease of the initial value within 14 days of exposure. The pass level for ready biodegradability (70% removal of DOC in a 10-day window within a 28-day period) was reached.
Degradation in the Toxicity Control:
In the toxicity control, containing the test item (corresponding to 50.1% of total DOC), the reference item (corresponding to 49.9% of total DOC) and activated sludge (inoculum), the initial DOC concentration of 58.4 mg/L measured on Day 0 decreased to 0.5 mg/L on Day 14. Biodegradation amounted to 99% within 14 days of exposure. Thus, according to the test guidelines, the test item was not inhibitory to activated sludge at the tested concentration of 50.4 mg/L because degradation was > 35% within 14 days.
Measurement of pH and Temperature:
The pH measured in all flasks at the end of exposure (Day 28) was 7.2 to 7.4. The temperature during the test period was in the range 21-24°C.
Table: Dissolved Organic Carbon Concentration (DOC) in the Test Flasks
Time [days] |
DOC [mg/L]* |
|||||||||
Test Item |
Procedure control |
Inoculum control |
Toxicity control |
|||||||
Replicate No. |
Replicate No. |
Replicate No. |
Replicate No. |
|||||||
1 |
2 |
mean# |
1 |
2 |
mean# |
1 |
2 |
mean# |
1 |
|
0 |
32.4 |
32.5 |
28.7 |
32.5 |
32.7 |
28.8 |
3.7 |
3.9 |
3.8 |
58.4 |
2 |
29.9 |
31.4 |
28.4 |
2.8 |
2.8 |
0.6 |
2.1 |
2.4 |
2.3 |
29.4 |
5 |
30.4 |
31.0 |
28.5 |
2.8 |
3.0 |
0.7 |
2.2 |
2.3 |
2.3 |
29.1 |
8 |
30.4 |
31.2 |
28.0 |
3.2 |
3.4 |
0.5 |
2.9 |
2.7 |
2.8 |
27.8 |
11 |
27.9 |
20.7 |
21.4 |
3.2 |
3.6 |
0.5 |
2.9 |
2.9 |
2.9 |
26.3 |
14 |
3.7 |
3.6 |
0.3 |
3.3 |
3.8 |
0.2 |
3.5 |
3.3 |
3.4 |
0.5 |
21 |
2.9 |
3.4 |
0.2 |
2.9 |
2.8 |
-0.1 |
2.8 |
3.0 |
2.9 |
0.4 |
28 |
3.5 |
3.8 |
0.3 |
3.9 |
3.9 |
0.6 |
3.9 |
2.8 |
3.4 |
0.8 |
* Mean values of at least triplicate measurements per sample.
# Values corrected for the mean inoculum control.
Table: Percentage Biodegradation in the Test Flasks
Time [days] |
Percentage Biodegradation* |
||||||
Test Item |
Procedure control |
Toxicity control |
|||||
Replicate No. |
Replicate No. |
Replicate No. |
|||||
1 |
2 |
mean |
1 |
2 |
mean |
1 |
|
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 |
3 |
-2 |
1 |
98 |
98 |
98 |
50 |
5 |
2 |
0 |
1 |
98 |
97 |
98 |
50 |
8 |
3 |
1 |
2 |
99 |
98 |
98 |
52 |
11 |
13 |
38 |
25 |
99 |
98 |
98 |
55 |
14 |
99 |
99 |
99 |
100 |
98 |
99 |
99 |
21 |
100 |
98 |
99 |
100 |
100 |
100 |
99 |
28 |
99 |
98 |
99 |
98 |
98 |
98 |
99 |
* Corrected for the inoculum controls.
Table: pH Values at the Start and at the End of the Test
Replicate No. |
Identification |
pH |
pH |
|
|
Start |
End |
1 |
Test item |
7.4 |
7.4 |
2 |
Test item |
7.4 |
7.4 |
1 |
Procedure control |
7.4 |
7.3 |
2 |
Procedure control |
7.4 |
7.3 |
1 |
Inoculum control |
7.4 |
7.2 |
2 |
Inoculum control |
7.4 |
7.2 |
1 |
Toxicity control |
7.4 |
7.3 |
Description of key information
Biodegradation in water (Cyrene™/Gem Diol): 99% in 28 days (DOC removal, OECD 301A)
Key value for chemical safety assessment
- Biodegradation in water:
- readily biodegradable
- Type of water:
- freshwater
Additional information
A biodegradation value of 99% in 28 days (DOC removal) was determined for the substance in accordance with OECD Test Guideline 301A and in compliance with GLP. The result is considered to be reliable. Since in aqueous media the Gem Diol predominates, the biodegradation observed is considered to reflect that Gem Diol is readily biodegradable.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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