Study initiation Study Title Data Requirements

I Test Guidelines **~·** ·--- Author: Study completion End of Experimental Sponsor: Testing facility: Laboratory Project Identification Study No. Page
1 of 22Study No.: This pageis intentionally left blank for the purpose of submitting administrative information that is required by regulations promulgated by various countries. Page 2 of 22CONTENTS GLP DECLARATION ARCHIVATION
OF DOCUMENTS AND TEST ARTICLES QUALITY ASSURANCE STATEMENT GLP CERTIFICATE SUMMARY INTRODUCTION EXPERIMENTAL PROCEDURE Test item Test organisms Pre-treatment

of the inoculum Pre-treatment

of the testItem Pre-treatment

of the reference compound Pre-treatment

of the toxicity control Exposure conditions Chemical analysis Applied SOPs and methods RESULTS Comments DEGRADATION CURVES Page 3of 22 Study No.: Page 4 4 6 7 8 9 10 11 12 12 14 14 14 14 15 15 16 21 22Study No.: GLP DECLARATION This study was conducted

in compliance with the OECD principles
of Good Laboratory **Practice~**and with the

of Good Laboratory Practice according
to **-~Chemical**

Law

· Date

I Signature Study Director ARCHIVATION
OF DOCUMENTS AND TEST ARTICLES Study plans, raw data and study reports are archived at the GLP archives of the test facility. Test articles are archived The sponsor must ensure that information submitted about the test item (identity testing or information about physical and chemical properties)

is complete and that it complies with the principles of Good Laboratory Practice. The customer

of the test item used where sampling has not been performed
Page 4 of 22

I !·· i I I I I I I I I I i I I i !Study No. This pageis intentionally left blank for the purpose of submitting administrative information that is required by regulations promulgated by various countries. Page 5 of 22Study No.: QUALITY ASSURANCE STATEMENT This report was audited Quality Management that the final report The dates of Quality Assurance inspections and audits are given below. During the on- site inspections procedures applicable
to this type of short time study were inspected. Audits study plan review process based inspection final report review Date:

Dates ofQA inspection Dates of report ·- **.~**.---- -· Quality Assurance Unit: Page 6 of 22Please note:
GLP CERTIFICATE GLP-Bnchelnlgu.ng/St•t•ment

of GLP Compllanc• (gemla/accordlng

to 11tb Ab».
1 Chemlkallenguetz} Elne **GlP~**zur 0beTWachUng det EJr\halll.mg cMr AUtUmtlll
or C'Oflfonntly Mth GUO IIC'COI'dl11-'
10 GLP·Grundfltuo gernU Chemikallengnetz hrw. RlcNinie **r:Mmlhlkn~**:.d Olmttw IU3201CEC

1t : 8&1320iEGwurde durchg.tiltwt
In: Pr!ifungen nach Katogorien Areas
o f EJq>ertise **(;end~.**OlemYW

·GLP Nt. 5.3o«CO gt,ifanct) ltce«dkki ChemllwV.Q,.
,...,5.31'0f.CO **~)** Kategorie

1
category

1 Prfifungen zur Sest immung der **ph~lltalltch-**physlcal-dlemlcal tMii'lg ehetnischen Eigensdlaft•l und Geheltsbesllmnungen Kategono

4
eategory

-4 Okoloxikologlsche Prllfungen zur Bestlmmung der environmental toxicity tluclles on aqooUc ond Auswlr1c:unoe" auf aqualische und letrestriseh.e t.,..trlal ot;anlsma Organ! **~men** ""'-

5
catogo<y

5 PrQfungen zum Verhalten 1m Boden, IM Wasser und In studies on behavtour

In water, sol and air; dar Luh: PrOfungen **n.~r**Bloakkumuletion und zur bloacc;umulatlon Melabolisrerung KArl_.,

8 colegoryl Analytrsthe PrUfungen an bio{ogischen Uaterlalien analytlcal and dlnlcal chemlstty tesllng Datum der lnspektion

Date of Inspection **~**

.

.. .**-·---~·** Study No.: Die geoannte Pt\'tfek'lrtchtung beftndM Sleh 1m natr GLP-Oberwlld\unoSwrlahren

und -Mtd IIIQIIIImiB,iy •uf GLP Complianl.:a PfOUtllnlftMI .,u;S Is lnspect.d on I **,.g~ar** E inhal' tung derGLP·GtuMsllze Oberwecht. bests. Auf der GtuncLJge des **~'fidl~**Md hivm¥\. B;,.w.ll.KttiwiMp«::onNpartlteanbetonflrrned

,th attia be.stlllgt. da» In dleser PrOfelnrk::htunu
die Dben **tettfaciJtyllebleto~lhe•foremenlionedstuc:lesin** genann.ten PrQfungen unter Einr'oltung

·der GLP

· toft'C)I

I anoo wtlb tho Prtndpktaot GLP. Grundsltl' **du~.rotvt**Mrden kOnnen. Page 7 of 22SUMMARY to assess the Study No

.:

of Inherent operating Regulation (EC) No in a mineral medium was inoculated with 40 % sludge of

cipal sewage treatment plants (STP) and 20 % sludge of an industrial STP and incubated under aerobic conditions. During this period, degradation was followed by continuous automated BOD determinations. Therefore,

86 % degradation after
7 days 1 00 % degradation after 14 days 100% degradation after 21 days 1 00% degradation after 27* days considered
to be inherently biodegradable. The reference compound sodium benzoate showed 68 % degradation after 7 days and 67% degradation after 14 days. * By mistake, programming

of the automated BOD-meter (OxiTop bottles) ended one day earlier than foreseen

in the study plan (i.e. on day 27). The test duration

is within the limits given in the guideline (14-28d) and, as 100% biodegradation were observed, has no influence on the overall study result. There
is indication of biodegradation, and clearly above the pass level for the test item of 70 %. Page 8 of 22Study No.: INTRODUCTION to assess the

of The study was conducted

in accordance with the OEC **edified**MITI **~**(based on standard CD Guideline _.-ancf
the Council Regulation (EC) No **~method**._"Manometric Respirometry Test"-· with fresh inocula **~nt**sources. A measured volume of inoculated mineral medium, containing

a known concentration

of 26.2 mg active ingredient (a.i.)/L of the test item was stirred in a closed flask at a constant temperature (25 ± 2 oc) for up to 28 days. The consumption

of oxygen (BOD) was determined
by measuring the drop in pressure

in the automated OxiTop respirometer flask. Evolved carbon dioxide was absorbed

in sodium hydroxide. The amount of oxygen taken up by the test item (corrected for uptake by blank inoculum, run in parallel) was expressed as a percentage
of chemical oxygen demand (COD). Page 9 of 22Study No. EXPERIMENTAL PROCEDURE A suspension
of 26.2 mg (a.i.)/L test item in amineral medium was inoculated and incubated under aerobic conditions

in the dark. The endogenous activity of the inoculum was checked running parallel blanks with inoculum but without test item. A reference compound (sodium benzoate) wasrunin parallel to check the operation
of the procedures. A toxicity control (test item and reference compound mixed) wasrunin parallel, to ensure that the chosen concentration

of the test item wasnot inhibitory
to microorganisms. Degradation was followed by the determination

of oxygen uptake and measurements were taken at frequent intervals to allow the identification
of the beginning and end of biodegradation. The test lasted 28 days, but programming

of the automated BOD-meter (OxiTop bottles) ended one day earlier than foreseen
in the study plan (i.e. on day 27). Because of the nature of biodegradation and of the mixed bacterial populations used as inoculum, determinations

of test item and inoculum blank were carried out in triplicate, for the reference compound

in duplicate. The oxygen uptake was calculated from the readings taken at regular and frequent intervals, using the method given by the manufacturer

of the equipment.
At the end of incubation, the pH of the contents of the flasks was measured. A correction of the oxygen consumed by nitrification wasnot necessary, because of the determination

of the chemical oxygen demand of the test item. Start of the incubation period

: End of the incubation period

: Page 10 of 22Study MATERIALS AND METHODS Test item Test item (Product name): Drying residue Content of active ingredient Partie Charge Expiry date Appearance Storage Data of active ingredient Chemical name CAS number Empirical formula Structural formula Molecular weight Stability of test concentrations during exposure

: 35.05% 31.94% liquid room temperature 376.3 g/mol Page 11 of 22Study No.: MATERIALS AND METHODS Test organisms Type Origin Date of collections Concentration

of inoculum (dry weight) Pre-treatment

of the inoculum mixed population
of aquatic microorganisms (activated sludge) 2 x40% activated sludge of an aeration tank of two different wastewater treating **predominant~** domestic
water authority ... sewage treatment plant treating predominantly sewage of industrial origi

.

---....,_. --

**~** ]._

. 100 mg/L suspended solids (ss) for all treatments with test item and toxicity control and 30 mg/L (ss) in the reference compound, respectively -**t~e**three sludges were **~ixed**proportionally ¥Is described before ¥

. -the sludge was washed twice by adding mineral medium and centrifuging for 1 0 min at 2000 rpmand 20oc and decanting off the supernatant -then the sludge wasfirst dissolved

in mineral medium and afterwards stored at room temperature under continuous shaking with aeration until usefor two**~ays**

.; **:~**

\f • ' 1.076 gof the wet sludge were **d~ied**in orderW calculate

;the amount of wet sludge to achieve

a concentration

of activated sludge of·5 g/1:. (ss) Page 12 of 22Study No.

: MATERIALS AND METHODS Mineral medium The mineral medium was prepared from stock solutions of mineral components described as follows. 1. Mineral salt solution 2 .Magnesium sulphate solution 3. Calcium chloride solution 4. Iron (Ill) chloride solution Potassium phosphate, monobasic (KH2P04) 8.5 g di-Potassium hydrogen phosphate (K2HP04) 21.75
g di-Sodium hydrogen phosphate dodecahydrate (Na2HP04

·12 H20)

44.6 g Ammonium chloride (NH4CI)

1.7 g The components were dissolved
in deionised water and filled up to 1 litre. The pH of the solution was adjusted to 7 .4. Magnesium sulphate heptahydrate (MgS04

·7 H20)
22.5g The substance was dissolved
in deionised water and filled up to 1 litre. Calcium chloride dihydrate (CaCI2

·2 H20)

36.4 g The substance was dissolved
in deionised water and filled up to 1 litre. Iron (Ill) chloride hexahydrate (FeCia

·6 H20)
0.25g The substance was dissolved
in deionised water and filled up to 1 litre. To prepare the mineral medium

3 ml each of the mineral salt solution

,the magnesium sulphate-, calcium chlo
ri de- and the iron (Ill) chloride solution are mixed and made up to 1000 ml with deionised water. Page 13 of 22Study No.: MATERIALS AND METHODS Pre-treatment
of the test item 20.5 mg of the test item were weighed out on piece of plastic. Both were added into the test flasks, which had already been filled with 200 ml of mineral medium. Afterwards the flask content was made up to 250 ml with mineral medium containing inoculum to give a test concentration

of 26.2 mg test item (a.i.)/L. Pre-treatment
of the reference compound Reference compound name

: sodium h""''" 7 "'"'td Purity

: 100.1% Batch-no.

:- 25 mg of the reference compound were weighed out on aluminium foil. Both were added into the test flasks, which had already been filled with 200 ml of mineral medium. Afterwards the flasks were made up to 250 ml with mineral medium containing inoculum
to give a test concentration

of 1 00 mg reference compound/L. Pre-treatment
of the toxicity control 20.5 mg of the test item and 7
.5 mg of the reference compound were weighed out on piece of plastic

I aluminium foil, respectively. Both were added to the test flasks, which had already been filled with 200 ml of mineral medium. Afterwards the flask content was made up to 250 ml with mineral medium containing inoculum
to give a test concentration

of 26.2 mg/L test item and 30 mg/L reference compound. Exposure conditions Test volume Test apparatus Mixing Incubation time Incubation temperature 250ml OxiTop System (WTW) 1 magnetic stirrer per test vessel 28 days 25 ± 2 oc Page 14 of 22Chemical analysis COD Method

Study No.: MATERIALS AND METHODS Determination

of Chemical Oxygen Demand of solids and substances which arenot watersoluble (COD). Calibration Standard Test apparatus Potassium hydrogen phthalate Titrator

I Methrohm 7301759 Applied SOPs and methods SOP

I Method 00090 Version

1 2011-0619601-070 Deviations Function Manom. Respirometry Test Determination

of Chemical Oxygen Demand of solids and substances In deviation
to the SOP 00090 Version
1, in all treatments with test item the test was conducted with 100 mg/L (dry weight) sludge inoculum of three origins (see test organisms) and 26.2 mgll test item. The mineral medium used is that of the "Modified MITI Test (II)" (OECD 302C) and is described in the report. All resulting deviations are documented

in the rawdata and in the report. Page 15 of 22Table 1: RESULTS pH-values after 28 days flask no. 1 2 3 4 5 6 7 8 9 Study No. pH-value 7.2 7.3 7.3 8.3 8.5 6.4 6.5 6.4 6.7 Page 16 of 22Study No.. RESULTS Table 2: BOD values (mg 02/L) after n days blankreference toxicity testitem blank mean compound control flask no.

6 7 8 1

2
3 4 5 9 code*
a1 a2 a3 b1 b2 b3 bm
r1 r2
t1 time (d) 0

0

0
0
0
0

00.0

0

0

0 1 -5.5 9.7-8.3 -2.8 -1.4 -5.5
0.0 26.3 19.4-4.1 2 -1.4 19.4 -5.5 2.84.11.4 2.8 44.2 45.6
-1.4 3 5.5 35.9-1.4 6.9 6.9 5.5 6.4 101 1054.1 4 22.1 48.4 6.9 9.712.4 9.7 10.6 11511915.2 5 27.7 58.1 18.0 15.2 15.2 15.2 15.2 119 12620.7 6 34.6 66.4 24.9 18.0 18.0 18.0 18.0 126131 26.3 7 37.3 73.3 29.022.1 20.7 20.721.2 135135 30.4 8 42.9 78.8 34.6 24.923.5 23.524.0 136140 34.6 9 48.4 85.7 38.7 30.4 27.727.7 28.6 138144 38.7 10 56.7 89.9 42.9 31.829.0 29.029.9 13814744.2 11 62.2 92.6 45.6 33.2 30.4 30.431.3 139148 49.8 12 65.0 96.8 49.8 37.333.2 34.6 35.0 14015155.3 13 69.1 99.5 55.3 40.1 34.6 35.9 36.9 146152 56.7 14 71.9 101 59.5 38.735.9 35.9 36.8 14515358.1 15
71

.910263.6 41.5 35.9 37.3 38

.2 146153 59.5 16 74.7 10466.4 42.9 37.3 40.1 40.1 14715662.2 17 70.5 10165.0 40.1 33.2 35.9 36.4 150152 59.5 18 74.7 10667.745.6 37.3 41.5 41.5 148 15662.2 19 78.8 108 69.1 45.6 40.1 42.9 42.9 149 15865.0 20 76.0 105 69.1 45.6 37.3 41.5 41.5 148 156 65.0 21 77.4 10670.5 48.4 38.7 44.2 43.8 151 15866.4 22 78.8 106 70.5 49.838.7 45.6 44.7 151 158 66.4 23 81.6 10873.3 49.840.1 45.6 45.2 152 16066.4 24 83.0 10973.3 51.2 41.547.0 46.6 153160 69.1 25 83.011173.3 52.5 41.548.4 47.5 151 15967.7 26 85.7 11174.7 53.942.9 49.8 48.9 152 160 69.1 27 88.511174.7 53.942.9 49.8 48.9 152160 69.1 Comments: *Abbreviations according
totest guideline. Page 17 of 22RESULTS Table3: Degradation
oftestitem Testconcentration (a.i.) Chemical oxygendemand (analytically determined) correctedBOD testitem (a-bm) mg02/L) time (d) a1a2 a3 00.00.00.0 10.09.70.0 20.016.60.0 30.029.50.0 411.537.80.0 512.542.92.8 616.648.46.9 716.152.17.8 818.954.810.6 919.857.110.1 1026.860.013.0 1130.961.314.3 1230.061.814.8 1332.262.618.4 1435.164.222.7 1533.763.825.4 1634

.663.926.3 1734.164.628.6 1833.264.526.2 1935.965.126.2 2034.563.527.6 2133.662.226.7 2234.161.325.8 2336.462.828.1 2436.462.426.7 2535.563.525.8 2636.862.125.8 2739

.668.732.4 Comments:none
26.2 mg/L 0.524 mg02/mg % degradation a1a2a3 0
0

0 071

0 0100

0 0100

0 84100

0 9110020 10010050 10010057 10010077 10010074 10010094 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 100100100 Page18of22 StudyNo.: % degradation mean 0 24 33 33 61 71 84 86 93 91 98 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100Study N RESULTS Table 4: Degradation
of reference compound Test concentration

1 00 mg/L Empirical formula

C1 Hs Na 02 Molecular weight
144.1 g/mol Theoretical oxygen demand 1.665 mg 0/mg corrected BOD reference compound

% degradation

% degradation (r-bm)

mean (mg 02/L) time (d)
r1
r2

r1

r2 0 0.0 0.0

0

0

0 1 26.3 19.4 16 12

14 2 41.4 42.8 25 26

25 3 94.6 98.6 57 59

58 4 104.4 108.4
63
65

64 5 103.8 110.8
62
67

64 6 108.0 113.0
65
68

66 7 113

.8 113

.8 68
68

68 8 112.0 116.0
67
70

68 9 109.4 115.4
66
69

68 10 108.1 117.1
65
70

68 11 107.7 116.7
65
70

67 12 105.0 116.0
63
70

66 13 109

.1 115.1
66
69

67 14 108.2 116.2
65
70

67 15 107.8 114.8
65
69

67 16 106.9 115.9
64
70

67 17 113.6 115.6
68
69

69 18 106.5 114.5

64
69

66 19 106.1 115.1

64
69

66 20 106.5 114.5

64
69

66 21 107.2 114.2

64
69

67 22 106.3 113.3

64
68

66 23 106.8 114.8
64
69

67 24 106.4 113.4

64
68

66 25 103.5 111

.5 62
67

65 26 103.1 111.1
62
67

64 27 103.1 111.1

62
67

64 Comments: none Page 19 of22RESULTS Table 5: Toxicity control Test concentration test item (a.i.) Test concentration reference compound corrected BOD toxicity control (t-bm)
% degradation (mg Oz/L) time (d)

t1

t1 0

0.0 1

0.0 2

0.0 3

0.0 4

4.6 5

5.5 6

8.3 7

9.2 8

10.6 9

10.1 10

14.3 11

18.5 12

20.3 13

19.8 14

21.3 15

21.3 16

22.1 17

23.1 18

20.7 19

22.1 20

23.5 21

22.6 22

21.7 23

21.2 24

22.5 25

20.2 26

20.2 27

20.2
Study 26.2 mg/L 30 mg/L 0 0 0 0 7 9 13 15 17 16 22 29 32 31 33 33 35 36 33 35 37 36 34 33 35 32 32 32 Comments: The used concentrations

of the test item did not show toxic effects to bacteria. Page 20 of 22Study No.: RESULTS ation of 100 °/o was determined for s considered
to be "Inherently The degradation curves are given in figure 1. Comments The validity criteria of the test method were met: The percent degradation
of the reference compound sodium benzoate reached the level of<!: 40% after 7 days and<!: 65% after 14 days. No toxicity of the test item was observed

in the toxicity control. * By mistake, programming

of the automated BOD-meter (OxiTop bottles) ended one day earlier than foreseen

in the study plan (i.e. on day 27). The test duration

is within the limits given in the guideline (14-28d) and, as 100% biodegradation were observed, has no influence on the overall study result. There
is indication of biodegradation, and clearly above the pass level for the test item of 70 %. Page 21 of 22:fl. II :s c" 0 **~** "' ., Q 100 90 80 70 60 50 40 30 20 10

Study No. DEGRADATION CURVES 0 1 2 3 4 5 6 7 8 9

10 11 12 13 14 15 16 17 18 19

20 21 22 23 24 25 26 27 28 Tlme,[t]"d Fig. 1 Degradation curves compound sodium n.on..,.,,,..,,1to toxicity control and reference Page 22 of22