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Environmental fate & pathways

Biodegradation in water: screening tests

Administrative data

Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16/04/2009 to 14/05/2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in accordance with recognised guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))
Deviations:
no
Principles of method if other than guideline:
Not applicable.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
No further information required.
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable.

Study design

Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge, adapted
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure):
- Laboratory culture: Activated sludge was collected, prepared and controlled by Chemicals Evaluation and Research Institute
- Method of cultivation: The activated sludge, which was cultivated for 18.5 hours after it had been added the synthetic sewage (glucose, peptone and potassium dihydrogenphosphate were dissolved in purified water, and the pH of the solution was adjusted to 7.0±1.0), was used.
- Storage conditions: At a temperature of 25±1°C under the conditions of darkness.
- Storage length: 28 days
- Preparation of inoculum for exposure: 3 mL of solutions A, B, C and D were made up to 1000 mL with purified water (Japanese Pharmacopeia, Takasugi Pharmaceutical Co., Ltd.), and then the pH of this solution was adjusted to 7.0.
- Concentration of sludge: Concentration of suspended solid in the activated sludge was 3280 mg/L.
- Water filtered: No
Duration of test (contact time):
28 d
Initial test substance concentration
Initial conc.:
100 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimation
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
TEST CONDITIONS
- Composition of medium: Not specified
- Additional substrate: Each of the test solutions were made up to 1000 mL with purified water.
- Solubilising agent (type and concentration if used): n/a
- Test temperature: 25±1°C
- pH: 7.0
- pH adjusted: Yes
- Aeration of dilution water: Not specified
- Suspended solids concentration: The activated sludge was added to each test vessel so that the concentration of the suspended solid reached 30 mg/L.
- Continuous darkness: Yes

TEST SYSTEM
- Culturing apparatus: Closed system oxygen consumption measuring apparatus
- Number of culture flasks/concentration: 3
- Method used to create aerobic conditions: Not specified
- Method used to create anaerobic conditions: n/a
- Measuring equipment: Closed system oxygen consumption measuring apparatus:
- Temperature controlled bath and measuring unit : Asahi Techneion Co., Ltd.
- Data sampler : Asahi Techneion Co., Ltd.
- Vessel: 300 mL in volume (Improved type)
- Absorbent for carbon dioxide: Soda lime No.1 (for absorption of carbon dioxide, Wako Pure Chemical Industries, Ltd.)
- Test performed in closed vessels due to significant volatility of test substance: No
- Test performed in open system: Not specified

SAMPLING
- Sampling frequency: Not specified
- Sampling method: Not specified
- Sample storage before analysis: Not specified

CONTROL AND BLANK SYSTEM
- Inoculum blank: In one test vessel, nothing was added to the basal culture medium

STATISTICAL METHODS:
-The concentration of DOC (Dissolved Organic Carbon) in the sample for TOC was calculated by subtracting concentration of the inorganic carbon (IC) from concentration of the total carbon (TC). The concentration of TC and IC in the test solutions was proportionally calculated from the peak area of the test solution by comparison with that of 80.0 mgC/L standard solution for TC analysis and 80.0 mgC/L standard solution for IC analysis, respectively.
-The concentration of the test item in the sample for HPLC analysis was calculated proportionally by comparing the peak area on the chromatogram of the sample for HPLC analysis with that on the chromatogram of 101 mg/L standard solution.

-Percentage biodegradation by BOD:

Percentage biodegradation (%) = (BOD - B/TOD) x 100

BOD = Biochemical oxygen demand in the test solution
B = Biochemical oxygen demand in the control blank
TOD = Theoretical oxygen demand required when the test item was completely oxidized

Percentage biodegradation by DOC:

Percentage biodegradation (%) = (DOCw - DOCs/DOCw) x 100

DOCs = Residual amount of the dissolved organic carbon in the test solution
DOCw = Residual amount of the dissolved organic carbon in the test solution

Percentage biodegradation of test item:

Percentage biodegradation (%) = (Sw - Ss/Sw) x 100

Ss = Residual amount of the test item in the test solution
Sw = Residual amount of the test item in the test solution



Reference substance
Reference substance:
aniline

Results and discussion

Preliminary study:
Not performed
Test performance:
The concentration of DOC corresponding to the minimum determination limit was regarded as 1.0 mg C/L.
The lowest detectable peak area of the test item was regarded as 50000 µV • sec considering the noise level, which corresponded to the test item concentration of 0.42 mg/L.
% Degradation
Parameter:
% degradation (O2 consumption)
Value:
0 - 4
Sampling time:
28 d

BOD5 / COD results

BOD5 / COD
Parameter:
BOD5
Value:
0 - 2.2 mg O2/g test mat.
Results with reference substance:
Biodegradation of Analine at 28 days was 77%

Any other information on results incl. tables

The test item is a mixture which consists of many components 15310 ingredients, as such it was detected as two peaks that could divide on the HPLC chromatogram in the determination of the test item. The two peaks were referred to as peak 1 and 2 in elution order. The percentage biodegradation of each peak of the test item was in the range from 2 to 3%. From these results, it was considered that all of the components of the test item were not biodegraded by microorganisms under the test conditions of this study.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Interpretation of results:
under test conditions no biodegradation observed
Conclusions:
The test item was not biodegraded by microorganisms under the test conditions of this study.
Executive summary:

In a biodegradation study of Ink BH11 C by microorganisms (CERI study number: 15304) the test material was not considered to have biodegraded under the conditions of the study.

The method followed that described in the OECD Guidelines for Testing of Chemicals, Guideline 301 C, Revised July 17, 1992,

Ready Biodegradability: Modified MITI Test (I).

The test item is a mixture which consists of many components 15310 ingredients, as such it was detected as two peaks that could divide on the HPLC chromatogram in thedetermination of the test item. The two peaks were referred to as peak 1 and 2 in elution order. The percentage biodegradation of each peak of the test item was in the range from 2 to 3%. From these results, it was considered that all of the components of the test item were not biodegraded by microorganisms under the test conditions of this study.