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Biodegradation in water: screening tests

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Endpoint:
biodegradation in water: inherent biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: ISO 5815 Second Edition 1989-08-01 (E) corresponding to 92 / 96 / EEC C.5
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Code number: FAT 20036/F
Batch-No: 1
Purity: ca. 97.7 %
Appearance: Solid
Solubility: 80 mg/L (in water)
Storage: room temperature
Expiration date: 07/99
Oxygen conditions:
aerobic
Inoculum or test system:
other: Seeding water of an urban waste water
Details on inoculum:
Seeding water of an urban waste water
Duration of test (contact time):
5 d
Details on study design:
- Experimental conditions
- Test concentration: about 6 to 800 mg test substance / litre dilution water
- Temperature: 20 ± 1 °C
- Duration: 5 days
- Estimation Technique O2: - Determination measured with an oxygen sensitive electrode system : SYLAND (SAV : 00 107)
- Suppression of nitrification : without suppression of nitrification
Reference substance:
other: D(+) - Glucose (Merck Nr. 8337) / L-Glutamic acid (Merck Nr. 291)
Preliminary study:
No data
Test performance:
No data
Parameter:
other: BOD5
Remarks on result:
other: BOD5
Remarks:
Detemination of BOD5
Details on results:
No data
Key result
Parameter:
BOD5
Value:
0 mg O2/g test mat.
Results with reference substance:
Reference substance: The BOD5 of Glucose/Glutamic acid solution is 185mg O2/L

BOD values of the oxygen measurements:

 ml Ref. solution/litre solution water Oxygen conc. at day 0 in mg O2/L    Oxygen conc. at day 5 in mg O2/L  Oxygen consumed mg O2/L    Blank mg O2/L BOD5 mg O2/gr 
 20.0  8.7  4.6  4.1  0.5  180
 20.0  8.7  4.2  4.5  0.5  200
 20.0  8.7  4.7  4.0  0.5  175

BOD 5 = 185 mg O2/L

  mg Ref. solution/litre solution water   Oxygen conc. at day 0 in mg O2/L    Oxygen conc. at day 5 in mg O2/L    Oxygen consumed mg O2/L    Blank mg O2/L  BOD5 mg O2/gr 
 801.5   8.8  7.6  1.2  0.5  1
 801.5   8.8  7.5  1.3   0.5  1
 400.8   8.8  8.0  0.8   0.5  1
 400.8   8.8  8.1  0.7   0.5  0
 200.4  8.8  8.3  0.5   0.5  0
 200.4  8.7  8.2  0.5   0.5  0
 100.2   8.7  8.2  0.5   0.5  0
 100.2  8.7  8.2  0.5   0.5  0
 50.1  8.7  8.4  0.3   0.5  0
 50.1  8.7  8.5  0.2   0.5  0
 25.0   8.7  8.4  0.3   0.5  0
 25.0   8.7  8.4  0.3   0.5  0
 12.5   8.7  8.2  0.5   0.5  0
 12.5   8.7  8.1  0.6   0.5  8
 6.3   8.7 8.3   0.4   0.5  0
 6.3  8.7  8.0  0.7   0.5  32

BOD5 = 0 mg O2/gr.

Validity criteria fulfilled:
yes
Interpretation of results:
other: Biological Oxygen Demand
Conclusions:
The BOD5 of FAT - 20'036/F is 0 mg/g O2
Executive summary:

The biochemical oxygen demand of FAT - 20'036/F is determined by measuring the dissolved oxygen concentration before and after 5 days incubation at 20°C in different test concentrations diluted in water saturated with dissolved oxygen that contains a seed of microorganisms.

The test was carried out according to ISO 5815 Second Edition 1989-08-01 (E) corresponding to 92/96/EEC C.5. The test was performed in compliance with the Good Laboratory Practice (GLP) Regulations of Switzerland.

The test substance was tested in concentrations from about 6 to 800 mg/l. The reference substance D(+)-Glucose / L-Glutamic acid was tested in a concentration of 20 ml/l.

Result: The BOD5 of FAT - 20'036/F was found to be 0 mg 02/g

The BOD5 of the reference substance was found to be 185 mg O2/I

Endpoint:
biodegradation in water: screening tests
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: DEV DIN 38'409 - H 41-1 (1980) corresponding to 92/96/EEC C.6
Deviations:
yes
Remarks:
Mean of 2 detremination instead of 3
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Code number: FAT 20036/F
Batch-No: 1
Purity: ca. 97.7 %
Appearance: Solid
Solubility: 80 mg/L (in water)
Storage: room temperature
Expiration date: 07/99
Oxygen conditions:
aerobic
Inoculum or test system:
not specified
Details on inoculum:
Not applicable
Duration of test (contact time):
2 h
Details on study design:
Sample weight: 10 mg of the test substance
Temperature: 148 ± 3 °C
Duration: 2 hours
Estimation technique: Titration (potentiometric) METROHM Titroprozessor 670 (SAV: 00 111)
Reference substance:
other: Potassium hydrogen phthalate solution (Merck-Nr. 11'769)
Preliminary study:
Not applicanle
Test performance:
Not applicanle
Parameter:
other: COD
Remarks on result:
other: COD
Remarks:
Determination of COD
Details on results:
None
Key result
Parameter:
COD
Value:
493 mg O2/g test mat.
Results with reference substance:
Reference substance:
The COD of Potassium hydrogen phthalate is 201 mg O2/L
(Validity: 200 ± 8 mg O2/L)

COD Analytical data.

 Product Sample weight E (mg)  Titration (mL) a    Titration (mL) b    Titration (mL) c   N=  mg O2/g
 Normality       10.099 10.113  0.1188, 0.1186 Mean: 0.1187  
 Blank  

 9.908, 9.907

Mean 9.908

       
 FAT 20036/F  10.0 mg    4.672, 4.755    

497, 489

Mean:493 

                   
 Reference  Stock solution E (mL)    Titration ml b      mg O2/L
 Potassium Hydrogen phtalate 20 ml    5.6185.701     

 203, 199

Mean: 201

Validity criteria fulfilled:
yes
Interpretation of results:
other: Chemical Oxygen Demand
Conclusions:
The COD of FAT 20036/F is found to be 493 mg O2/g.
Executive summary:

A test was carried out according to DEV DIN 38'409 - H 41 -1 (1980) corresponding to 92/96/EEC C.6 to determine COD of FAT 20036/F. The test was performed in compliance with the Good Laboratory Practice (GLP) Regulations of Switzerland.

A predeterminated amount of the test substance FAT - 20'036/F dissolved in water is oxidized by potassium dichromate in a strong sulphuric acid medium with silver sulphate as a catalyst under reflux for 2 hours. The residual dichromate is determined by titration with standardized ferrous ammonium sulphate.

The test substance was tested with a sample weight of 10.0 mg. The reference substance, Potassium hydrogen phthalate solution was tested in a concentration of 20 ml/I.

The COD of FAT - 20036/F was found to be 493 mg 02/g The COD of the reference substance was found to be: 201 mg O2/L. Taking the results of the corresponding BOD5 test into account, the rsulting BOD5/COD quotient is zero and therefore no biodegradation can be expected.

Endpoint:
biodegradation in water: inherent biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 302 C (Inherent Biodegradability: Modified MITI Test (II))
Deviations:
yes
Remarks:
Duration: 21 days. Instead of 28 days, the test will be finished after 21 days since no degradation activity could be observed.
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Code number: FAT 20036/F
Batch-No: 1
Purity: ca. 97.7 %
Appearance: Solid
Solubility: 80 mg/L (in water)
Storage: room temperature
Expiration date: 07/99
ThOD: 1.42 mg 02/mg
Oxygen conditions:
aerobic
Inoculum or test system:
other: Mixture of polyvalent bacteria.
Details on inoculum:
- Mixture of polyvalent bacteria collected on August 09, 1994 in the aeration tank of a domestic sewage treatment plant, PRO RHENO Basle.
- Bacteria concentration: 100 mg/l suspended solids.
Duration of test (contact time):
21 d
Initial conc.:
30 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
O2 consumption
Details on study design:
Water / Nutrient: According to the specification of the test method
Temperature: 25 ± 2 °C
Test duration: 21 days
Lighting: none
Estimation Technic: Voith Sapromat; 02-consumption automatically measure in milligrams oxygen per liter.
Reference substance:
other: Natriunbenzoat used at 100 mg/l
Preliminary study:
No data
Test performance:
No data
Key result
Parameter:
% degradation (O2 consumption)
Value:
0
Sampling time:
21 d
Remarks on result:
other: The biodegradation of FAT - 20'036/F after 21 days corrected by the inoculum control and measured as BOD versus ThOD
Details on results:
- Test substance: The biodégradation of FAT - 20036/F after 21 days corrected by the inoculum control and measured as BOD versus ThOD is 0 %
- Abiotic control: The abiotic degradation at the end of the test and measured as BOD versus ThOD is 0 %
Parameter:
BOD5
Value:
0 mg O2/g test mat.
Results with reference substance:
The biodégradation of the reference substance corrected by the inoculum control and measured as BOD versus ThOD is :
- 77 % after 7 days
- 80 % after 14 days
- 82 % after 21 days

Oxygen consumption values:

                              Oxygen consumption in mg/l
         Inoculum reference        Reference substance           Test substance
 Days  Flask 1 Flask 2  Mean  Flask 3  Flask 4  Mean  Flask 17  Flask 18  Flask 19  Mean 
 2  26  28  27  120  129 124.5   18  21  20  19.7
 4  39  42  40.5  154  162 158  28  32  32  30.7
 5*  41  46  43.5  162  172  167  32  36  36  34.7
 6  43  50  46.5  166  178  172  34  38  38  36.7
 8  45  52  48.5  172  186  179  37  42  41  40
 10  47  56  51.5  176  191  183.5  40  45  44  43
 12  49  60  54.5  180  195  187.5  43  49  47  46.3
 14  52  62  57  182  198  190  44  51  49  48
 16  52  64  58  184  200  192  45  52  50  49
 18  52  66  59  186  202  194 46   53  51  50
 20  52  66  59  188  205  196.5  47  54  52  51
 21  52  68  60  190  205  197.5  48  54  52  51.3

MITI (II) TEST SAPROMAT

            Oxygen consumption in mg/L Biodegradtion in %    
 Days  Inoculum mean  ReferenceSubstance(100 mg/l)(Mean) Testsubstance(30 mg/l)(Mean)   Abioticcontrol(30 mg/l)Flask 20  Test substance Reference substance 
 2  27  124.5  19.7  0  0  58
 4  40.5  158  30.7  0  0  70
 5*  43.5  167  34.7  0  0  74
 6  46.5  172  36.7  0  0  75
 8  48.5  179  40  0  0  78
 10  51.5  183.5  43  0  0  79
 12  54.5  187.5  46.3  0  0  80
 14  57  190  48  0  0  80
 16  58  192  49  0  0  80
 18 59   194  50  0  0  81
 20  59  196.5  51  0  0  82
 21  60  197.5  51.3  0  0  82

Calculation of the BOD5:

When the test substance is not soluble in water and the determination of the BOD5 under the conditions of ISO 5815 is not possible, the BOD5 of the test substance is given by the equation:

BOD * (mg O2 / mg Test substance) =

((mg O2/l uptake by Test substance) - (mg O2/l uptake by blank)) (1) / mg/l Test substance X 1000

1) BOD Measurement at day 5

BOD5 = 0 mg 02/g

Validity criteria fulfilled:
yes
Interpretation of results:
under test conditions no biodegradation observed
Conclusions:
Biodegradation after 21 days of the test substance of FAT - 20'036/F is: 0 %
Executive summary:

The inherent biodegradability of FAT - 20'036/F was determined in a 21 days MITI (II) - Test according to the OECD Guideline for Testing of Chemicals, No. 302C, Paris 1981. The test was performed in compliance with the Procedures and Principles of Good Laboratory Practice (GLP)

The test substance was tested in a concentration of 30 mg/l. The reference substance (Natriumbenzoat) was tested in a concentration of 100 mg/I.

Result:

Biodegradation after 21 days of the test substance of FAT - 20'036/F is: 0 %

Biodegradation after 21 days of the reference substance is: 82 %

FAT - 20'036/F is not inherently biodegradable. From the results of the toxicity control, no toxic effect of the biodégradation of the reference substance is obvious.

Description of key information

Three independant key studies, according to OECD Guideline and GLP were performed to determined COD, BOD5 and the inherent biodegradability of FAT 20036/F.

The COD was found to be 493 mg O2/gr, BOD5 to be 0 mg O2/gr and the inherent biodegradbility to be 0%.

Based on theBOD5/CODquotient which is not >0.5, the substance is not indicative of rapid degradation.

Taking the data from degradation test and BOD5/COD quotient into account, the test substance FAT 20036 is considered to be not biodegradable.

Supporting studies were performed on different batches (A, B, C, D and E) to determine BOD, COD and biodegradtion values.

- FAT 20036/A: COD is found to be 994 mg/ gr O2, BOD5 to be 130 mg/gr O2 and the inherent biodegradability to be 69% after 31 days.

- FAT 20036/B:COD is found to be 1085 mg COD/ gr, BOD5 to be 17 mg/gr O2.

- FAT 20036/C:COD is found to be 811 mg/ gr O2, BOD5 to be 0 mg/gr O2.

- FAT 20036/D:COD is found to be 757 mg/ gr O2, BOD5 to be 16 mg/gr O2.

- FAT 20036/E: COD is found to be 807 mg/ gr O2, BOD5 to be 250 mg/gr O2 and the inherent biodegradability to be 83% DOC after 28 days.

The inherent biodegradability test and the BOD5/COD ratio (<0.5) indicate that the test substance FAT 20036 is not expected to be readily biodegradable.

Key value for chemical safety assessment

Biodegradation in water:
inherently biodegradable

Additional information

By the nature of their design and use, acid dyes are not intended to be readily biodegradable as this would assist in the rapid destruction of the dyestuff, rendering it unfit for purpose. As such, it is accepted that such substances are not readily biodegradable under relevant environmental conditions. A published study (Pagga & Brown, 1986) describes the results of the testing of 87 dyestuffs in short-term aerobic biodegradation tests. The authors of this publication concluded that dyestuffs are very unlikely to show any significant biodegradation in such tests and that 'there seems little point in carrying out such test procedures’ on dyestuffs.

There are ISO, European, American (AATCC) and national standards for the colour fastness of dyes. Dyes are required to have specific fastness properties. If the dyes were biodegradable, it would not be possible for them to have these fastness properties.