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Biodegradation in water and sediment: simulation tests

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Reference
Endpoint:
biodegradation in water: simulation testing on ultimate degradation in surface water
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 May 2020 to 18 Sep 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 309 (Aerobic Mineralisation in Surface Water - Simulation Biodegradation Test)
Version / remarks:
2004
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Radiolabelling:
no
Oxygen conditions:
aerobic
Inoculum or test system:
natural water
Details on source and properties of surface water:
An overview of the Sampling Site, Sampling Conditions and Water Characteristics is provided in Table 1 in 'Any other information on materials and methods incl. tables'.
- Details on collection: The water was collected from the surface of the river 'Leine' (approx. upper 20 cm water) at an undisturbed recess. Furthermore, temperature, pH value and O2 concentration of the water were measured at field sampling.
- Location of collection: Am Anger, 30982 Pattensen, Germany
- Temperature at time of collection: 12.2 °C
- pH at time of collection: 7.64
- Oxygen concentration: 9.43 mg O2/L
- Dissolved organic carbon: 5.21 mg/L
- Handeling: The water was clear and colourless. Small fishes were removed by filtration.
- Suspended mater: To minimize possible interferences of NER formation with biodegradation and to allow proper kinetic evaluation, water with an amount of suspended matter < 10 mg/L was used.
- Acclimation: Until test start the surface water was acclimated under test conditions at 12 ± 2 °C in the dark with aeration.
- Storage: Storage at 4 ± 2 °C was not necessary as the test was started within 2 days after collection of the water.
Details on source and properties of sediment:
Not applicable
Details on inoculum:
Not applicable
Duration of test (contact time):
90 d
Initial conc.:
20 µg/L
Based on:
test mat.
Initial conc.:
100 µg/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
other: The amount of test item constituents and degradation products was determined
Details on study design:
TEST CONDITIONS
- Volume of test solution/treatment: 80 mL (test item replicates, sterile controls and sterile blank controls); 100 mL (blank controls); 150 mL (reference control and solvent control)
- Test temperature: 12 ± 2 °C
- pH: 7.50 – 7.85
- Oxygen concentration: 9.29 – 9.60 mg O2/L
- Agitation: Continuous stirring at approx. 120 rpm to maintain particles and microorganisms in suspension and to facilitate oxygen transfer from the headspace to the liquid.
- Continuous darkness: Yes

TEST SYSTEM
- Culturing apparatus: 120 mL gastight glass bottles for test item replicates, sterile controls and sterile blank controls. 150 mL gastight glass bottles for blank controls. 300 mL gastight bottles for reference control and solvent control replicates.
- Number of replicates for test item: 2/sample/ concentration (in total 28 replicates were prepared for each concentration)
- Number of replicates for sterile controls: 2/sample/ concentration (in total 16 replicates were prepared for each concentration)
- Number of replicates for blank controls: 1 (in total 10 replicates were prepared for each concentration)
- Number of replicates for sterile blank controls: 1 (in total duplicates were prepared)
- Number of replicates for reference control: 2
- Number of replicates for solvent control: 2

SAMPLING
- Temperature: The incubation temperature was recorded continuously throughout the test.
- pH and Oxygen: Measurements of pH and oxygen concentration in the test system (Blank control) were done on day 0, 33, 40, 62 and 89.
- Transformation: Sampling for determination of the transformation was carried out directly after application and at 13 additional sampling points. The sampling points were chosen to enable the establishment of the pattern of decline of the test item constituents. Sampling was done after application (day 0) and at day 2, 5, 8, 12, 14, 20, 33, 43, 51, 57, 62, 75 and day 90. For the reference control sampling was stopped once degradation was finished. Sampling was done at day 0, day 6 and day 12. Residual activity in the test solution and dissolved 14CO2 (after acidification of test solution) was determined by LSC.
- Sampling method used per analysis type: The amount of test item constituents and degradation products in the test solution and sterile test item solution was determined by LC-HRMS. Two replicates per test item concentration were harvested at each sampling time for determination of transformation until day 62. At day 75 and day 90 the remaining four replicates were analysed. Sterile controls were analysed in parallel at each sampling point. One blank control and one sterile blank control were analysed in parallel to the respective sampling.

Reference substance:
other: Aniline sulfate, [14C(U)]-
Remarks:
10 μg/L
Compartment:
natural water: freshwater
% Recovery:
92
Remarks on result:
other: Constituent 9; 20 µg/L
Compartment:
natural water: freshwater
% Recovery:
96
Remarks on result:
other: Constituent 10; 20 µg/L
Compartment:
natural water: freshwater
% Recovery:
90
Remarks on result:
other: Constituent 6; 20 µg/L
Compartment:
natural water: freshwater
% Recovery:
95
Remarks on result:
other: Constituent 1; 20 µg/L
Compartment:
natural water: freshwater
% Recovery:
94
Remarks on result:
other: Constituent 7; 20 µg/L
Compartment:
natural water: freshwater
% Recovery:
96
Remarks on result:
other: Constituent 2; 20 µg/L
Compartment:
natural water: freshwater
% Recovery:
88
Remarks on result:
other: Constituent 3; 20 µg/L
Compartment:
natural water: freshwater
% Recovery:
87
Remarks on result:
other: Constituent 4; 20 µg/L
Compartment:
natural water: freshwater
% Recovery:
89
Remarks on result:
other: Constituent 5; 20 µg/L
Compartment:
natural water: freshwater
% Recovery:
100
Remarks on result:
other: Constituent 9; 100 µg/L
Compartment:
natural water: freshwater
% Recovery:
100
Remarks on result:
other: Constituent 10; 100 µg/L
Compartment:
natural water: freshwater
% Recovery:
100
Remarks on result:
other: Constituent 6; 100 µg/L
Compartment:
natural water: freshwater
% Recovery:
85
Remarks on result:
other: Constituent 1; 100 µg/L
Compartment:
natural water: freshwater
% Recovery:
99
Remarks on result:
other: Constituent 7; 100 µg/L
Compartment:
natural water: freshwater
% Recovery:
100
Remarks on result:
other: Constituent 2; 100 µg/L
Compartment:
natural water: freshwater
% Recovery:
100
Remarks on result:
other: Constituent 3; 100 µg/L
Compartment:
natural water: freshwater
% Recovery:
100
Remarks on result:
other: Constituent 4; 100 µg/L
Compartment:
natural water: freshwater
% Recovery:
100
Remarks on result:
other: Constituent 5; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
100
Parameter:
test mat. analysis
Sampling time:
62 d
Remarks on result:
other: all studied constituent; 20 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
99
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 9; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
98
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 10; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
99
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 11; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
95.8
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 6; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
94
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 1; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
95.2
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 7; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
95.2
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 2; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
95.2
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 3; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
95.2
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 4; 100 µg/L
Parent/product:
parent
Compartment:
water
% Degr.:
95.2
Parameter:
test mat. analysis
Sampling time:
90 d
Remarks on result:
other: Constituent 5; 100 µg/L
Key result
Compartment:
natural water: freshwater
DT50:
25.2 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 5; 20 μg/L
Key result
Compartment:
natural water: freshwater
DT50:
37.3 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 1; 100 μg/L
Compartment:
natural water: freshwater
DT50:
19.6 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: Constituent 9; 20 μg/L
Compartment:
natural water: freshwater
DT50:
21.2 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: Constituent 10; 20 μg/L
Compartment:
natural water: freshwater
DT50:
19.4 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: Constituent 6; 20 μg/L
Compartment:
natural water: freshwater
DT50:
21.8 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 7; 20 μg/L
Compartment:
natural water: freshwater
DT50:
19.6 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 2; 20 μg/L
Compartment:
natural water: freshwater
DT50:
22.5 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 3; 20 μg/L
Compartment:
natural water: freshwater
DT50:
24 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 4; 20 μg/L
Compartment:
natural water: freshwater
DT50:
13.9 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 9; 100 μg/L
Compartment:
natural water: freshwater
DT50:
13.7 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 10; 100 μg/L
Compartment:
natural water: freshwater
DT50:
11.9 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 11; 100 μg/L
Compartment:
natural water: freshwater
DT50:
13.8 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 6; 100 μg/L
Compartment:
natural water: freshwater
DT50:
16.2 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 7; 100 μg/L
Compartment:
natural water: freshwater
DT50:
13.7 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 2; 100 μg/L
Compartment:
natural water: freshwater
DT50:
14.8 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 3; 100 μg/L
Compartment:
natural water: freshwater
DT50:
19.7 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 4; 100 μg/L
Compartment:
natural water: freshwater
DT50:
25.2 d
Type:
(pseudo-)first order (= half-life)
Temp.:
12 °C
Remarks on result:
other: constituent 5; 100 μg/L
Transformation products:
not specified
Remarks:
M4, M5, M9, M10, M11, M20, M13, M14, M17, M18 and M19
Details on transformation products:
An overview of the transformation products is provided in Table 12 - Table 13 in ‘Any other information on results incl. tables’.
Metabolites were formed by oxidation of the primary or secondary alcohol moieties of the constituents. Eleven metabolites (M) were identified in the test. The link between metabolites and corresponding constituents of the test item is list below:
M4 and M5 were identified in Constituent 6 transformation products
M9 was identified in Constituent 7 transformation products
M10 and M11 were identified in Constituent 2 transformation products
M13, M14 and M20 were identified in Constituent 3 transformation products
M17, M18 and M19 were identified in Constituent 4 transformation products

The kinetic evaluations for the metabolites were performed based on the FOCUS guidance document on estimating persistence and degradation kinetics. DTx values were calculated based on single first order kinetics (SFO), the metabolite data were normalized for these calculations. The highest occurrence of a metabolite was fixed at 100 % correspondingly the respective day was set to day 0. The results indicate that the DT50 values of the metabolites at 100 µg/L applied concentration in surface water were in a range of 7.57 to 14.4 days. An overview of the results is provided in Table 14 in ‘Any other information on results incl. tables’.
Evaporation of parent compound:
not specified
Volatile metabolites:
no
Residues:
yes
Details on results:
An overview of the results is provided in Table 4 – Table 11 in ‘Any other information on materials and methods incl. tables’.
The test substance cannot be radiolabelled, because it is a UVCB substance. Due to the use of an unlabelled test item, analysis of Non-extractable residues (NER) was not possible.

Transformation of most constituents started in within 5 – 12 days after application. About 50% transformation was reached within 33 - 43 days, after 62 days the detected concentration was < LOQ or even not detectable. For constituent 1 transformation started only on day 43 and the transformation reached 50% after 57 days and 65 % on day 62. At day 75 the concentration was < LOQ.
In the sterile controls, no significant decrease was determined until day 90 for the constituents 9, 10 and 11. For constituent 1 - 7 no significant decrease was determined until day 43, thereafter the concentration decreased slightly until day 90.
The difference in dissipation (transformation) in the biotic test item replicates and the abiotic sterile controls clearly indicate that biodegradation is the major pathway of dissipation (transformation).

The kinetic evaluations were done based on the FOCUS guidance document on estimating persistence and degradation kinetics.
The kinetic models were chosen based on the following criteria:
• Visual assessment of the fitted and observed data versus time
• Visual assessment of the residuals
• Estimation of the error percentage at which the x2-test was passed
Results with reference substance:
An overview of the results is provided in Table 3 in ‘Any other information on results incl. tables’
Mineralisation of the reference item Aniline sulphate, [14C(U)]- in the reference control was 60% after 6 days and 65.6% after 12 days. In the solvent control the mineralisation was 50 % after 6 days and 64% after 12 days. The high mineralization indicates that the surface water contained an active microbial population and the microbial activity was not inhibited by the solvent.

Table 3 Mineralization of Aniline Sulphate, [14C(U)]- in the Reference Control and Solvent Control

Exposure Day

Reference Control

Solvent Control

% Mineralisation1)

% Mineralisation1)

 

Repl. 1

Repl. 1

Repl. 2

Repl. 2

6

59.7

60.3

50.8

49.4

12

66.0

65.2

64.0

64.5

1) calculated from residual activity in test solution (after acidification) and applied radioactivity

Table 4. Recovery Rates Procedural Recovery Samples: 20 µg/L (Approach 2)

Fortified concentrations (n = 1 per sampling day): 2 x LOQ: 15 µg/L

Sampling Day

Constituent 9

Constituent 10

Constituent 6

Constituent 1

Constituent 7

Constituent 2

RR
[%]

RR
[%]

RR
[%]

RR
[%]

RR
[%]

RR
[%]

0

93

100

92

651)

90

95

2

84

87

86

81

83

92

5

96

110

97

88

96

96

8

81

93

87

112

82

97

12

95

107

100

107

103

96

14

101

89

95

85

95

88

20

91

88

96

88

97

97

33

95

107

113

146

122

107

43

86

91

86

79

84

90

51

85

92

89

85

87

94

57

88

96

95

108

96

91

62

92

96

90

95

94

96

Mean

84

89

87

87

87

88

Sampling Day

Constituent 3

Constituent 4

Constituent 5

 

 

 

RR
[%]

RR
[%]

RR
[%]

 

 

 

0

98

85

100

 

 

 

2

90

95

100

 

 

 

5

104

95

102

 

 

 

8

94

92

85

 

 

 

12

95

98

108

 

 

 

14

89

102

114

 

 

 

20

92

96

88

 

 

 

33

109

114

97

 

 

 

43

83

81

93

 

 

 

51

98

94

96

 

 

 

57

106

96

93

 

 

 

62

88

87

89

 

 

 

Mean

88

87

90

 

 

 

Table 5. Recovery Rates Procedural Recovery Samples: 100 µg/L (Approach 1)

Fortified concentrations (n = 1 per sampling day): 2 x LOQ: 15 µg/L

Sampling Day

Constituent 9

Constituent 10

Constituent 6

Constituent 1

Constituent 7

Constituent 2

RR
[%]

RR
[%]

RR
[%]

RR
[%]

RR
[%]

RR
[%]

0

120

135

118

128

112

126

2

100

115

100

83

100

99

5

102

107

97

106

95

114

8

93

108

98

113

100

105

12

103

112

98

108

103

94

14

90

107

106

70

102

101

20

90

101

102

82

104

97

33

82

95

88

75

98

88

43

99

105

87

97

84

77

51

85

86

91

104

98

105

57

126

97

95

88

95

100

62

124

101

107

119

99

108

75

108

89

113

91

111

111

90

113

101

103

85

99

106

Mean

103

104

100

96

100

102

Sampling Day

Constituent 3

Constituent 4

Constituent 5

 

 

 

RR
[%]

RR
[%]

RR
[%]

 

 

 

0

126

136

123

 

 

 

2

100

105

113

 

 

 

5

108

105

91

 

 

 

8

103

102

118

 

 

 

12

102

99

93

 

 

 

14

108

124

107

 

 

 

20

104

101

103

 

 

 

33

92

82

88

 

 

 

43

80

81

71

 

 

 

51

100

109

97

 

 

 

57

96

97

102

 

 

 

62

108

101

108

 

 

 

75

111

113

129

 

 

 

90

114

106

115

 

 

 

Mean

104

104

104

 

 

 

Table 6. 20 µg/L test item

 

 

% Applied (nominal)

Constituent

Repl.

Day

 

0

2

5

8

12

14

Constituent 9

1

112.1

115.6

113.3

96.2

108.0

85.9

2

104.6

112.2

105.4

104.4

85.1

58.1

Mean

108.4

113.9

109.3

100.3

96.6

72.0

Constituent 10

1

105.2

99.9

118.1

84.7

90.3

76.2

2

124.1

112.0

117.3

113.3

92.1

64.1

Mean

114.7

105.9

117.7

99.0

91.2

70.1

Constituent 6

1

87.5

77.1

71.3

62.5

68.2

57.9

2

90.3

77.7

77.9

73.2

62.4

48.8

Mean

88.9

77.4

74.6

67.9

65.3

53.3

Constituent 1

1

126.1

90.7

181.1

111.7

108.4

101.0

2

131.6

121.5

176.6

125.9

82.2

91.2

Mean

128.9

106.1

178.8

118.8

95.3

96.1

Constituent 7

1

85.3

73.1

67.9

56.8

70.6

63.2

2

92.8

73.0

87.1

75.7

57.5

50.7

Mean

89.1

73.1

77.5

66.3

64.1

57.0

Constituent 2

1

103.8

99.7

100.4

81.2

88.4

67.3

2

108.0

97.2

100.5

78.9

65.9

55.1

Mean

105.9

98.5

100.5

80.1

77.1

61.2

Constituent 3

1

95.6

93.9

91.0

74.9

76.0

75.4

2

96.7

88.7

92.4

81.5

71.1

60.4

Mean

96.2

91.3

91.7

78.2

73.6

67.9

Constituent 4

1

91.4

94.4

95.9

74.7

84.0

79.5

2

98.3

95.0

99.2

78.6

72.8

80.1

Mean

94.8

94.7

97.6

76.7

78.4

79.8

Constituent 5

1

98.9

100.8

87.5

74.3

87.3

88.6

2

101.6

96.5

94.1

90.7

75.5

62.5

Mean

100.2

98.7

90.8

82.5

81.4

75.5

 

Table 6 (continued). 20 µg/L test item

 

 

% Applied (nominal)

Constituent

Repl.

Day

 

20

33

43

51

57

62

Constituent 9

1

62.2

23.8

<LCL

<LCL

<LCL

<LCL

2

54.7

21.3

<LCL

<LCL

<LCL

n.d.

Mean

58.5

22.6

<LCL

<LCL

<LCL

<LCL

Constituent 10

1

65.0

<LCL

n.d.

n.d.

n.d.

n.d.

2

53.8

32.3

<LCL

n.d.

n.d.

n.d.

Mean

59.4

n.c.

<LCL

n.d.

n.d.

n.d.

Constituent 6

1

41.2

27.6

<LCL

<LCL

<LCL

<LCL

2

36.8

23.1

<LCL

<LCL

<LCL

<LCL

Mean

39.0

25.4

<LCL

<LCL

<LCL

<LCL

Constituent 1

1

81.5

141.2

57.0

<LCL

111.6

n.d.

2

113.1

105.3

92.5

<LCL

<LCL

<LCL

Mean

97.3

123.3

74.8

<LCL

n.c.

<LCL

Constituent 7

1

45.3

34.7

<LCL

<LCL

<LCL

<LCL

2

45.6

16.8

<LCL

<LCL

<LCL

<LCL

Mean

45.5

25.7

<LCL

<LCL

<LCL

<LCL

Constituent 2

1

53.4

39.1

<LCL

<LCL

<LCL

<LCL

2

56.3

24.4

n.d.

<LCL

<LCL

<LCL

Mean

54.8

31.7

<LCL

<LCL

<LCL

<LCL

Constituent 3

1

58.8

34.5

<LCL

<LCL

11.9

<LCL

2

60.7

22.8

n.d.

<LCL

<LCL

<LCL

Mean

59.7

28.6

<LCL

<LCL

n.c.

<LCL

Constituent 4

1

69.8

50.0

12.8

10.5

16.0

<LCL

2

74.0

33.7

16.0

<LCL

<LCL

<LCL

Mean

71.9

41.8

14.4

n.c.

n.c.

<LCL

Constituent 5

1

65.3

54.1

20.9

20.8

19.4

n.d.

2

71.0

43.9

19.4

22.4

<LCL

<LCL

Mean

68.1

49.0

20.1

21.6

n.c.

<LCL

Repl. = Replicate; LCL = Lowest calibration level; n.d. = not detected; n.c.= not calculable 

Table 7. 100 µg/L test item

 

 

% Applied (nominal)

Constituent

Repl.

Day

 

0

2

5

8

12

14

20

33

Constituent 9

1

95.9

93.5

93.0

96.7

63.4

60.0

33.4

4.8*

2

92.5

92.6

117.9

88.1

59.4

59.4

23.0

21.2

Mean

94.2

93.0

105.4

92.4

61.4

59.7

28.2

13.0

Constituent 10

1

94.3

92.6

89.5

96.0

61.6

54.0

27.3

9.5*

2

95.7

88.6

107.3

80.3

57.7

56.3

20.1

24.5

Mean

95.0

90.6

98.4

88.2

59.7

55.2

23.7

17.0

Constituent 11

1

97.0

101.9

94.5

84.9

59.4

52.0

21.8

4.8*

2

93.8

96.8

96.9

80.7

46.8

53.5

18.3

24.2

Mean

95.4

99.4

95.7

82.8

53.1

52.8

20.1

14.5

Constituent 6

1

86.3

78.8

73.2

75.6

46.4

42.8

26.3

9.9

2

83.1

81.6

81.7

65.2

40.9

44.8

17.9

28.5

Mean

84.7

80.2

77.5

70.4

43.6

43.8

22.1

19.2

Constituent 1

1

99.3

93.7

97.5

103.3

81.6

95.5

79.1

81.6

2

89.9

97.9

109.7

93.1

90.1

95.0

93.5

107.2

Mean

94.6

95.8

103.6

98.2

85.8

95.2

86.3

94.4

Constituent 7

1

84.0

79.5

72.9

74.7

53.1

48.6

30.9

15.0

2

83.7

83.3

77.9

65.6

44.7

50.7

22.5

32.5

Mean

83.9

81.4

75.4

70.1

48.9

49.6

26.7

23.8

Constituent 2

1

89.0

92.9

84.8

85.4

54.6

48.9

28.3

8.7

2

86.7

92.1

95.5

67.4

44.5

52.8

17.4

33.0

Mean

87.8

92.5

90.1

76.4

49.6

50.9

22.9

20.9

Constituent 3

1

90.4

90.5

77.7

84.7

57.1

52.9

31.9

14.3

2

85.6

89.0

91.1

67.6

49.5

53.8

19.2

31.1

Mean

88.0

89.8

84.4

76.1

53.3

53.3

25.5

22.7

Constituent 4

1

89.4

90.3

81.7

83.8

66.9

63.1

47.6

27.4

2

85.6

90.9

89.4

70.3

57.0

62.3

33.7

43.8

Mean

87.5

90.6

85.5

77.1

61.9

62.7

40.7

35.6

Constituent 5

1

91.9

87.2

78.9

82.9

70.4

76.2

59.3

43.7

2

85.9

85.6

91.0

70.3

61.0

66.5

45.2

60.9

Mean

88.9

86.4

84.9

76.6

65.7

71.3

52.3

52.3

Table 7 (continued).100 µg/L test item

 

 

% Applied (nominal)

Constituent

Repl.

Day

 

43

51

57

62

75

90

Constituent 9

1

4.8*

4.8*

4.8*

4.8*

4.8*

4.8*

1.0*

1.0*

2

4.8*

4.8*

4.8*

4.8*

9.1

4.8*

1.0*

1.0*

Mean

4.8

4.8

4.8

4.8

6.9

4.8

1.0

1.0

Constituent 10

1

9.5

2.0

2.0

2.0

9.5*

9.5*

2.0*

2.0*

2

9.5

2.0

2.0

2.0

12.0

9.5*

2.0*

2.0*

Mean

9.5

2.0

2.0

2.0

10.8

9.5

2.0

2.0

Constituent 11

1

1.0*

1.0*

1.0*

1.0*

1.0*

1.0*

1.0*

1.0*

2

1.0*

1.0*

1.0*

1.0*

25.8

1.0*

1.0*

1.0*

Mean

1.0

1.0

1.0

1.0

13.4

1.0

1.0

1.0

Constituent 6

1

13.0

4.8

6.0

4.8

4.8

4.8

4.8*

4.8*

2

10.1

4.8

4.8

4.8

12.8

7.2

4.8*

4.8*

Mean

11.6

4.8

5.4

4.8

8.8

6.0

4.8

4.8

Constituent 1

1

85.8

30.6

103.9

53.7

6.0*

6.0*

6.0*

6.0*

2

77.0

11.2

6.0

15.7

7.9

6.0*

6.0*

6.0*

Mean

81.4

20.9

55.0

34.7

7.0

6.0

6.0

6.0

Constituent 7

1

15.1

8.2

6.0

4.8*

6.4

4.8*

4.8*

4.8*

2

11.6

9.6

4.8*

4.8*

14.4

9.5

4.8*

4.8*

Mean

13.3

8.9

5.4

4.8

10.4

7.1

4.8

4.8

Constituent 2

1

13.1

4.8*

5.9

4.8*

4.8*

4.8*

4.8*

4.8*

2

6.3

4.8*

4.8*

4.8*

6.7

4.8*

4.8*

4.8*

Mean

9.7

4.8

5.3

4.8

5.7

4.8

4.8

4.8

Constituent 3

1

18.0

4.8*

8.7

4.8*

4.8*

4.8*

4.8*

4.8*

2

4.8*

4.8*

4.8*

4.8*

9.0

4.8*

4.8*

4.8*

Mean

11.4

4.8

6.7

4.8

6.9

4.8

4.8

4.8

Constituent 4

1

38.6

7.2

22.1

6.8

4.8*

4.8*

4.8*

4.8*

2

8.6

6.0

4.8*

4.8*

9.7

6.5

4.8*

4.8*

Mean

23.6

6.6

13.4

5.8

7.2

5.6

4.8

4.8

Constituent 5

1

54.8

15.7

33.4

13.1

6.0

4.8*

4.8*

4.8*

2

20.7

11.7

4.8*

4.8*

11.6

7.9

4.8*

4.8*

Mean

37.8

13.7

19.1

8.9

8.8

6.3

4.8

4.8

Repl. = Replicate    *= value < lowest calibration level or not detected, respectively     

Table 8. Sterile Control - 20 µg/L

 

 

% Applied (nominal)

Constituent

Repl.

Day

 

0

2

5

8

12

14

Constituent 9

1

102.8

121.3

103.0

125.1

132.1

136.7

2

84.4

109.7

129.5

106.9

118.5

111.9

Mean

93.6

115.5

116.3

116.0

125.3

124.3

Constituent 10

1

108.3

119.3

113.4

137.7

143.9

139.6

2

110.9

112.8

138.7

110.7

121.5

114.0

Mean

109.6

116.0

126.0

124.2

132.7

126.8

Constituent 6

1

88.2

107.9

78.8

92.8

95.7

84.8

2

90.6

98.7

103.0

87.2

88.7

76.5

Mean

89.4

103.3

90.9

90.0

92.2

80.6

Constituent 1

1

127.5

124.3

120.1

137.3

131.8

124.1

2

126.9

104.6

122.8

122.2

97.0

98.9

Mean

127.2

114.4

121.4

129.7

114.4

111.5

Constituent 7

1

90.2

105.2

77.4

82.4

95.9

70.2

2

91.4

95.9

101.7

77.2

88.2

68.6

Mean

90.8

100.5

89.5

79.8

92.1

69.4

Constituent 2

1

103.9

118.8

90.7

107.4

119.0

116.6

2

106.6

115.0

112.3

100.4

107.3

89.9

Mean

105.2

116.9

101.5

103.9

113.2

103.2

Constituent 3

1

105.4

120.3

84.7

98.7

116.5

87.6

2

100.6

111.2

106.2

98.8

95.3

77.8

Mean

103.0

115.8

95.4

98.8

105.9

82.7

Constituent 4

1

96.8

117.9

78.0

88.3

95.6

78.2

2

99.9

112.3

96.3

95.5

91.9

75.8

Mean

98.3

115.1

87.2

91.9

93.8

77.0

Constituent 5

1

100.8

121.6

69.0

77.7

104.6

61.6

2

105.2

116.7

89.3

86.5

78.0

53.2

Mean

103.0

119.1

79.2

82.1

91.3

57.4

 

Table 8 (continued). Sterile Control - 20 µg/L

 

 

% Applied (nominal)

Constituent

Repl.

Day

 

20

33

43

51

57

62

Constituent 9

1

109.9

137.5

124.1

104.4

103.5

115.5

2

111.8

132.0

122.0

103.6

110.6

96.3

Mean

110.8

134.8

123.1

104.0

107.0

105.9

Constituent 10

1

102.9

141.1

107.0

99.4

105.3

108.7

2

115.5

122.7

102.2

123.8

126.4

115.9

Mean

109.2

131.9

104.6

111.6

115.8

112.3

Constituent 6

1

78.2

73.5

64.6

53.9

74.0

58.6

2

115.9

70.3

47.2

69.9

75.6

65.8

Mean

97.0

71.9

55.9

61.9

74.8

62.2

Constituent 1

1

85.8

146.9

91.1

164.7

138.0

129.5

2

127.2

143.6

79.5

142.1

124.8

131.4

Mean

106.5

145.3

85.3

153.4

131.4

130.4

Constituent 7

1

78.0

74.6

55.0

53.9

64.2

49.3

2

117.2

51.7

31.6

57.4

66.3

61.6

Mean

97.6

63.2

43.3

55.6

65.3

55.4

Constituent 2

1

99.8

107.8

81.9

100.1

111.2

107.3

2

146.0

87.7

61.5

100.8

108.4

104.4

Mean

122.9

97.7

71.7

100.4

109.8

105.8

Constituent 3

1

92.8

96.5

60.5

85.3

91.0

88.2

2

126.1

71.8

32.1

81.4

96.0

87.9

Mean

109.4

84.2

46.3

83.4

93.5

88.1

Constituent 4

1

80.1

85.4

38.3

73.1

68.9

71.9

2

130.7

55.9

24.4

67.5

78.4

66.9

Mean

105.4

70.6

31.3

70.3

73.6

69.4

Constituent 5

1

71.1

71.7

28.3

63.4

52.1

55.6

2

110.8

37.4

20.0

69.3

56.9

67.2

Mean

91.0

54.6

24.2

66.4

54.5

61.4

Repl. = Replicate 

Table 9. Sterile Control - 100 µg/L

 

 

% Applied (nominal)

Constituent

Repl.

Day

 

0

2

5

8

12

14

20

33

Constituent 9

1

94.8

97.8

101.4

103.0

94.6

105.2

106.9

106.3

2

95.0

96.7

98.9

99.7

96.1

108.9

104.1

113.7

Mean

94.9

97.2

100.1

101.4

95.4

107.0

105.5

110.0

Constituent 10

1

94.1

103.8

94.0

95.8

92.0

101.4

100.3

105.5

2

98.6

98.9

97.4

101.2

99.3

109.0

101.7

115.0

Mean

96.4

101.4

95.7

98.5

95.6

105.2

101.0

110.2

Constituent 11

1

92.2

101.5

92.9

95.9

87.7

100.6

99.3

115.2

2

104.8

94.5

106.0

93.6

98.3

93.2

105.8

105.6

Mean

98.5

98.0

99.4

94.8

93.0

96.9

102.6

110.4

Constituent 6

1

81.8

84.0

81.6

80.6

79.7

78.7

89.8

70.6

2

84.5

80.6

84.2

91.4

79.9

75.8

82.8

70.7

Mean

83.2

82.3

82.9

86.0

79.8

77.3

86.3

70.7

Constituent 1

1

105.1

101.2

103.9

107.7

96.4

101.3

106.8

114.6

2

99.8

97.3

97.4

104.2

105.2

101.0

110.8

106.0

Mean

102.5

99.2

100.6

106.0

100.8

101.1

108.8

110.3

Constituent 7

1

79.7

83.3

78.0

74.2

74.3

71.1

88.3

66.2

2

80.8

77.8

85.0

90.9

77.6

66.1

76.0

63.0

Mean

80.3

80.5

81.5

82.6

76.0

68.6

82.2

64.6

Constituent 2

1

94.9

104.4

93.6

95.9

97.6

94.9

111.3

92.1

2

92.4

101.3

100.2

103.8

94.2

93.3

103.2

86.1

Mean

93.7

102.8

96.9

99.9

95.9

94.1

107.2

89.1

Constituent 3

1

88.9

103.5

88.6

85.4

89.6

85.7

102.7

82.3

2

90.5

91.3

91.8

99.3

91.5

79.4

96.4

76.9

Mean

89.7

97.4

90.2

92.3

90.6

82.5

99.6

79.6

Constituent 4

1

89.3

99.3

85.3

77.7

85.5

78.3

94.7

76.9

2

90.8

91.5

92.5

94.9

81.8

69.0

85.3

72.5

Mean

90.0

95.4

88.9

86.3

83.7

73.6

90.0

74.7

Constituent 5

1

86.0

94.1

89.2

77.4

80.6

67.9

90.3

74.1

2

86.7

87.0

99.8

97.8

80.9

58.4

81.4

66.1

Mean

86.3

90.6

94.5

87.6

80.8

63.1

85.9

70.1

Table 9 (continued). Sterile Control - 100 µg/L

Constituent

Repl.

% Applied (nominal)

Day

43

51

57

62

75

90

Constituent 9

1

98.8

97.5

97.6

97.9

91.2

88.4

2

103.0

99.9

100.6

102.9

114.5

93.6

Mean

100.9

98.7

99.1

100.4

102.9

91.0

Constituent 10

1

101.0

95.0

102.9

103.6

92.7

77.8

2

98.5

98.7

102.4

105.7

114.6

95.1

Mean

99.7

96.8

102.6

104.6

103.7

86.5

Constituent 11

1

98.2

97.9

91.5

102.5

68.4

77.5

2

115.0

98.9

110.8

100.0

120.7

102.8

Mean

106.6

98.4

101.2

101.3

94.6

90.2

Constituent 6

1

72.4

46.8

46.0

36.5

29.2

20.4

2

80.6

59.8

59.6

59.6

58.2

50.2

Mean

76.5

53.3

52.8

48.1

43.7

35.3

Constituent 1

1

98.8

101.6

90.7

68.4

63.6

50.3

2

117.3

104.7

75.7

91.9

89.4

87.2

Mean

108.1

103.1

83.2

80.1

76.5

68.8

Constituent 7

1

67.9

40.1

39.0

30.4

20.8

15.9

2

73.2

58.0

54.6

54.9

53.1

44.8

Mean

70.5

49.0

46.8

42.6

36.9

30.3

Constituent 2

1

97.5

69.8

75.8

68.1

48.0

34.3

2

104.6

79.8

82.6

89.4

70.7

66.2

Mean

101.0

74.8

79.2

78.7

59.3

50.3

Constituent 3

1

90.9

61.0

62.9

54.7

39.6

31.1

2

95.8

71.1

70.1

70.8

57.6

56.4

Mean

93.3

66.0

66.5

62.7

48.6

43.8

Constituent 4

1

79.1

58.0

55.5

49.8

39.8

29.9

2

82.9

65.2

59.6

60.5

54.5

49.6

Mean

81.0

61.6

57.5

55.2

47.1

39.7

Constituent 5

1

74.4

52.9

52.1

47.2

40.9

31.0

2

70.9

57.6

54.6

53.6

56.4

45.3

Mean

72.6

55.2

53.4

50.4

48.6

38.1

Repl. = Replicate

Table 10. Data of Kinetic Fit and DTx Values of the test item - 20 μg/L

Endpoint / Statistic

Single First Order (SFO)

Constituent

Constituent 9

Constituent 10

Constituent 6

Constituent 7

C0

122.6
± 6.179

121.6
± 5.657

88.34
± 2.389

86.68
± 3.881

Initial value for fitting

100

100

100

100

K1 (1/d)

0.03545
± 0.005252

0.03272
± 0.005029

0.03565
± 0.002825

0.03186
± 0.004459

Initial value for fitting

0.1

0.1

0.1

0.1

r2

0.842

0.8208

0.9435

0.8393

c2 error (parent)

9.47

7.82

4.64

5.30

 

DTx Values in days

DT50

19.6

21.2

19.4

21.8

Endpoint / Statistic

Single First Order (SFO)

Constituent

Constituent 2

Constituent 3

Constituent 4

Constituent 5

C0

109.0
± 3.504

100.8
± 2.810

104.3
± 4.366

105.5
± 3.51

Initial value for fitting

100

100

100

100

K1 (1/d)

0.03538
± 0.003349

0.03083
± 0.002613

0.02885
± 0.003258

0.02753
± 0.002415

Initial value for fitting

0.1

0.1

0.1

0.1

r2

0.9205

0.9462

0.8995

0.9281

c2 error (parent)

4.78

5.64

12.1

7.07

 

DTx Values in days

DT50

19.6

22.5

24.0

25.2

 

Table 11. Data of Kinetic Fit and DTx Values of the test item - 100 µg/L

Endpoint / Statistic

Single First Order (SFO)

Constituent

Constituent 9

Constituent 10

Constituent 11

Constituent 6

Constituent 1

C0

110.7
± 4.971

107.1
± 4.513

110.4
± 4.752

89.83
± 3.081

111.4
± 8.292

Initial value for fitting

100

100

100

100

100

K1 (1/d)

0.004996
± 0.004922

0.005047
± 0.004662

0.05832
± 0.005457

0.05007
± 0.003766

0.01857
± 0.003106

Initial value for fitting

0.1

0.1

0.1

0.1

0.1

r2

0.9343

0.9402

0.9432

0.9588

0.7066

c2 error (parent)

19.6

17.9

17.8

12.7

20.6

 

DTx Values in days

DT50

13.9

13.7

11.9

13.8

37.3

DT90

46.1

45.6

39.5

46

124

Endpoint / Statistic

Single First Order (SFO)

Constituent

Constituent 7

Constituent 2

Constituent 3

Constituent 4

Constituent 5

C0

88.18
± 2.564

99.75
± 3.903

97.27
± 3.336

65.80
± 3.173

94.28
± 3.747

Initial value for fitting

100

100

100

100

100

K1 (1/d)

0.04287
± 0.002735

0.05061
± 0.004342

0.04673
± 0.003519

0.03524
± 0.002539

0.02748
± 0.002356

Initial value for fitting

0.1

0.1

0.1

0.1

0.1

r2

0.9668

0.9471

0.9569

0.9522

0.9212

c2 error (parent)

9.96

14.8

12.2

9.18

10.9

 

DTx Values in days

DT50

16.2

13.7

14.8

19.7

25.2

DT90

53.7

45.5

49.3

65.3

83.8

 

Table12. Identified metabolites

Metabolite
(M)

Molecular
Formula

Retention Time
[min]

Accurate Mass
Adduct [M+H]+
m/z

Collision Energy
[V]

4

C26H43NO5

6.58

450.3214

20

5

C26H45NO5

6.53

452.3371

20

9

C29H51NO6

6.66

510.3789

20

10

C31H54N2O7

5.47

567.4004

20

11

C31H56N2O7

5.44

569.4160

20

20

C34H58N2O8

5.53

623.4266

20, 30

13

C34H60N2O8

5.51

625.4422

20

14

C34H62N2O8

5.48

627.4579

20, 30

17

C37H64N2O9

5.59

681.4685

30

18

C37H66N2O9

5.54

683.4841

30

19

C37H68N2O9

5.53

685.4998

30

Table 13. Link between Metabolites and corresponding Constituents of the Test Item

           DBE = Double Bond Equivalents

 

Constituent

Metabolite

 

No.

6

M4

M5

 

DBE

4

6

5

 

Mass

Fragment Ions [amu]

291.2319

289.2162
261.2213

291.2319

 

Fragment Ions

[C19H31O2]+

[C19H29O2]+
[C18H29O]+

[C19H31O2]+

 

No.

7

M9

--

 

DBE

4

5

--

 

Mass
Fragment Ions [amu]

291.2319
275.2369

289.2162

--

 

Fragment Ions

[C19H31O2]+

[C19H31O]+

[C19H29O2]+

--

 

 

Constituent

Metabolite

No.

2

M10

M11

--

DBE

4

6

5

--

Mass
Fragment Ions [amu]

466.3527
408.3108
303.2319

464.3371
303.2319

464.3371
408.3108
303.2319

--

Fragment Ions

[C27H48NO5]+
[C24H42NO5]+
[C20H31O2]+

[C27H46NO5]+
[C20H31O2]+

[C27H46NO5]+
[C24H42NO4]+
[C20H31O2]+

--

No.

3

M20

M13

M14

DBE

4

7

6

5

Mass
Fragment Ions [amu]

524.3946

466.3527
303.2319

464.3371
303.2319

464.3371
303.2319

466.3527
464.3371
303.2319

Molecular Formula Fragment Ions

[C30H54NO6]+ [C27H48NO5]+
[C20H31O2]+

[C27H46NO5]+
[C20H31O2]+

[C27H46NO5]+
[C20H31O2]+

C27H48NO5]+
[C27H46NO5+
[C20H31O2]+

No.

4

M17

M18

M19

DBE

4

7

6

5

Mass
Fragment Ions [amu]

524.3946
466.3527
303.2319

303.2319

522.3789
464.3371
303.2319

522.3789
303.2319

Fragment Ions

[C30H54NO6]+ [C27H48NO5]+ [C20H31O2]+

[C20H31O2]+

[C30H52NO6]+
[C27H46NO5]+
[C20H31O2]+-

[C30H52NO6]+ [C20H31O2]+

Table 14 Data of Kinetic Fit and DTxValues of Metabolites100 µg/L

Endpoint / Statistic

100 µg/L

Model

Single First Order (SFO)

Metabolite

M4

M5

M9

M10

M11

M13

C0

111.2
± 10.48

117.5
± 17.20

115.0
± 21.86

103.5
± 14.87

96.13
± 18.43

97.08
± 15.49

Initial value for fitting

100

100

100

100

100

100

K1 (1/d)

0.05278
± 0.008055

0.03289
± 0.007743

0.03432
± 0.01163

0.05671
± 0.01504

0.08724
± 0.03072

0.09153
± 0.02702

Initial value for fitting

0.1

0.1

0.1

0.1

0.1

0.1

r2

0.8539

0.6742

0.5601

0.7259

0.6278

0.7129

c2 error (parent)

23.6

29.5

35.7

19.6

27.7

21.1

 

DTx Values in days

DT50

13.1

21.1

20.2

12.2

7.95

7.57

DT90

43.6

70

67.1

40.6

26.4

25.2

Endpoint / Statistic

100 µg/L

 

Model

Single First Order (SFO)

 

Metabolite

M14

M17

M18

M19

M20

 

C0

111.1
± 19.09

99.01
± 11.64

98.12
± 12.67

110.5
± 18.34

110.2
± 18.39

 

Initial value for fitting

100

100

100

100

100

 

K1 (1/d)

0.05111
± 0.01475

0.08286
± 0.01779

0.08867
± 0.02108

0.04813
± 0.01347

0.05065
± 0.01596

 

Initial value for fitting

0.1

0.1

0.1

0.1

0.1

 

r2

0.6403

0.8188

0.7913

0.6472

0.6472

 

c2 error (parent)

28.8

12.8

14.8

23.1

24.4

 

 

 

 

DT50

13.6

8.37

7.82

14.4

13.7

 

DT90

45.1

27.8

26.0

47.8

45.5

 

Validity criteria:
The reference item aniline was degraded (> 50%) within 6 days.
Observed value:
The reference item aniline was degraded 60% within 6 days.
Validity criteria fulfilled:
yes
Conclusions:
In a biodegradation simulation study that was performed in accordance with OECD TG 309, the DT50 values of the test item at 20 µg/L applied concentration in surface water were in a range of 19.4 to 25.2 days. The DT50 values of the test item at 100 µg/L applied concentration in surface water were in a range of 11.9 to 37.3 days. Metabolites were formed by oxidation of the primary or secondary alcohol moieties of the constituents. Eleven metabolites (M) were identified in the test. The DT50 values of the metabolites of the test item at 100 µg/L applied concentration in surface water were in a range of 7.57 to 14.4 days.
Executive summary:

The aerobic mineralization of the test item in surface water was determined in a simulation biodegradation test over a test period of 90 days according to OECD TG 309 and in compliance with GLP criteria. The study was carried out at 12 ± 2 °C, pH 7.50 – 7.85, 9.29 – 9.60 mg O2/L oxygen concentration in darkness. Duplicate per sample per concentration for test item and sterile controls were included in the test. In addition, one blank control, one sterile blank control, two reference controls and two solvent controls (methanol) were included in the experimental set-up as well. Sampling for determination of the transformation was carried out directly after application and at 13 additional sampling points (day 2, 5, 8, 12, 14, 20, 33, 43, 51, 57, 62, 75 and day 90). The amount of test item constituents and degradation products in the test solution and sterile test item solution was determined by LC-HRMS. Residual activity in the test solution and dissolved 14CO2 (after acidification of test solution) was determined by LSC.

The test item is a UVCB, a reaction product of p-nonylphenol, formaldehyde and diethanolamine, propoxylated. Thus, it cannot be radiolabelled, in turn, analysis of Non-extractable residues (NER) was not possible. Eleven constituents (target compounds) were known. The results showed that transformation of most constituents started in within 5 – 12 days after application. About 50 % transformation was reached within 33 - 43 days, after 62 days the detected concentration was < LOQ or even not detectable. For constituent 1 transformation started only on day 43 and the transformation reached 50% after 57 days and 65 % on day 62. At day 75 the concentration was < LOQ. Metabolites were formed by oxidation of the primary or secondary alcohol moieties of the constituents. Eleven metabolites (M) were identified in the test. M4 and M5 were identified in Constituent 6 transformation products. M9 was identified in Constituent 7 transformation products. M10 and M11 were identified in Constituent 2 transformation products. M13, M14 and M20 were identified in Constituent 3 transformation products. M17, M18 and M19 were identified in Constituent 4 transformation products.

Based on the findings, the DT50 values of the test item at 20 µg/L applied concentration in surface water was in a range of 19.4 to 25.2 days. The DT50 values of the test item at 100 µg/L applied concentration in surface water were in a range of 11.9 to 37.3 days. The DT50 values of the metabolites of the test item at 100 µg/L applied concentration in surface water was in a range of 7.57 to 14.4 days.

Description of key information

The DT50 values of the test item at 20 µg/L applied concentration in surface freshwater were in a range of 19.4 to 25.2 days. The DT50 values of the test item at 100 µg/L applied concentration in surface freshwater were in a range of 11.9 to 37.3 days. The DT50 values of the metabolites of the test item at 100 µg/L applied concentration in surface freshwater were in a range of 7.57 to 14.4 days.

Key value for chemical safety assessment

Half-life in freshwater:
37.3 d
at the temperature of:
12 °C

Additional information

The aerobic mineralization of the test item in surface water was determined in a simulation biodegradation test over a test period of 90 days according to OECD TG 309 and in compliance with GLP criteria. The study was carried out at 12 ± 2 °C, pH 7.50 – 7.85, 9.29 – 9.60 mg O2/L oxygen concentration in darkness. Duplicate per sample per concentration for test item and sterile controls were included in the test. In addition, one blank control, one sterile blank control, two reference controls and two solvent controls (methanol) were included in the experimental set-up as well. Sampling for determination of the transformation was carried out directly after application and at 13 additional sampling points (day 2, 5, 8, 12, 14, 20, 33, 43, 51, 57, 62, 75 and day 90). The amount of test item constituents and degradation products in the test solution and sterile test item solution was determined by LC-HRMS. Residual activity in the test solution and dissolved 14CO2 (after acidification of test solution) was determined by LSC.

The test item is a UVCB, a reaction product of p-nonylphenol, formaldehyde and diethanolamine, propoxylated. Thus, it cannot be radiolabelled, in turn, analysis of Non-extractable residues (NER) was not possible. Eleven constituents (target compounds) were known. The results showed that transformation of most constituents started in within 5 – 12 days after application. About 50 % transformation was reached within 33 - 43 days, after 62 days the detected concentration was < LOQ or even not detectable. For constituent 1 transformation started only on day 43 and the transformation reached 50% after 57 days and 65 % on day 62. At day 75 the concentration was < LOQ. Metabolites were formed by oxidation of the primary or secondary alcohol moieties of the constituents. Eleven metabolites (M) were identified in the test. M4 and M5 were identified in Constituent 6 transformation products. M9 was identified in Constituent 7 transformation products. M10 and M11 were identified in Constituent 2 transformation products. M13, M14 and M20 were identified in Constituent 3 transformation products. M17, M18 and M19 were identified in Constituent 4 transformation products.

Based on the findings, the DT50 values of the test item at 20 µg/L applied concentration in surface water was in a range of 19.4 to 25.2 days. The DT50 values of the test item at 100 µg/L applied concentration in surface water were in a range of 11.9 to 37.3 days. The DT50 values of the metabolites of the test item at 100 µg/L applied concentration in surface water was in a range of 7.57 to 14.4 days.

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