# Annex B



# Spirotetramat

B.8 Fate and behaviour in the environment

Reviewed by: AGES, Austria

Peer – reviewed by: US-EPA, USA PMRA, Canada

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Contents:

B.8 Fate and behaviour in the environment..............

................................................... ...............5

B.8.1 Route and rate of degradation in soil ..............

................................................... ................10

B.8.1.1 Route of degradation (OECD Annex IIA, 7.1) .........

................................................... .........10
B.8.1.1.1 Aerobic degradation in soil (OECD Annex IIA, 7.1.1)

................................................... ..........10
B.8.1.1.2 Anaerobic degradation in soil (OECD Annex IIA, 7.1. 2)................................................. ........36
B.8.1.1.3 Soil photolysis (OECD Annex IIA, 7.1.3) ............

................................................... .................42
B.8.1.1.4 Summary on route of degradation in soil (laboratory ) .................................................. ..........50

B.8.1.2 Rate of degradation in soil (OECD Annex IIA, 7.2, I IIA, 9.1) .......................................... ...54
B.8.1.2.1 Aerobic degradation rate of the active substance at 20 °C (OECD Annex IIA, 7.2.1,

IIIA, 9.1.1) .......................................

................................................... .....................................54
B.8.1.2.2 Aerobic degradation of the active substance at 10 ° C (OECD Annex IIA, 7.2.2)...................58
B.8.1.2.3 Aerobic degradation of relevant metabolites at 20 ° C (OECD Annex IIA, 7.2.3) ...................59
B.8.1.2.4 Anaerobic degradation rate of the active substance (OECD Annex IIA, 7.2.4, IIIA,

9.1.2).............................................

................................................... .......................................73
B.8.1.2.5 Anaerobic degradation rate of relevant metabolites (OECD Annex IIA, 7.2.5) ......................73
B.8.1.2.6 Summary on the rate of degradation in laboratory so il studies ......................................... .....74

B.8.1.3 Field studies (OECD Annex IIA, 7.3, IIIA, 9.2).....

................................................... .............79
B.8.1.3.1 Soil dissipation testing (OECD Annex IIA, 7.3.1, II IA, 9.2.1) ......................................... .........79
B.8.1.3.2 Soil residue testing...............................

................................................... ..............................102
B.8.1.3.3 Soil accumulation testing on relevant soils........

................................................... ................102
B.8.1.3.4 Supplemental studies ...............................

................................................... .........................102
B.8.1.3.5 Summary on field dissipation studies ...............

................................................... .................115

B.8.2 Adsorption, desorption and mobility in soil ........

................................................... .........118

B.8.2.1 Mobility studies (OECD Annex IIA, 7.4, IIIA, 9.3) ..

................................................... ........118
B.8.2.1.1 Adsorption and desorption of the active substance ( OECD Annex IIA, 7.4.1) .....................118
B.8.2.1.2 Adsorption and desorption of relevant metabolites, degradation and reaction products

(OECD Annex IIA, 7.4.2) ............................

................................................... .......................123
B.8.2.1.3 Column leaching studies with the active substance ( OECD Annex IIA, 7.4.3, IIIA,

9.3.1).............................................

................................................... .....................................139
B.8.2.1.4 Column leaching studies with relevant metabolites ( OECD Annex IIA, 7.4.4, IIIA, 9.3.1)....139
B.8.2.1.5 Aged residue column leaching studies (OECD Annex II A, 7.4.5) ........................................15 0
B.8.2.1.6 Leaching (TLC) (OECD Annex IIA, 7.4.6) .............

................................................... ............150
B.8.2.1.7 Lysimeter studies (OECD Annex IIA, 7.4.7, Annex III A, 9.3.2) .......................................... ..150
B.8.2.1.8 Field leaching studies (OECD Annex IIA, 7.4.8, Anne x IIIA, 9.3.3)..................................... .150
B.8.2.1.9 Volatility laboratory and field study (OECD Annex I IA, 7.4.9, IIIA, 9.3.4, 9.3.5)...................15 0
B.8.2.1.10 Summary on the mobility in soil ....................

................................................... .....................151

B.8.3 Predicted environmental concentration in soil (PEC S 

### ) (OECD Annex IIIA, 9.4, 9.5) ....154

B.8.4 Fate and behaviour in the water....................

................................................... .................155

B.8.4.1 Hydrolytic degradation (OECD Annex IIA, 7.5) .......

................................................... ......155

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.4.2 Photolytic degradation (OECD Annex IIA, 7.6) .......

................................................... ......160

B.8.4.3 Ready biodegradability of the active substance (OEC D Annex IIA, 7.7).......................171

B.8.4.4 Degradation in aquatic systems (OECD Annex IIA, 7.8 ).................................................1 72
B.8.4.4.1 Aerobic biodegradation in aquatic systems (OECD Ann ex IIA, 7.8.1) .................................172
B.8.4.4.2 Anaerobic biodegradation in aquatic systems (OECD A nnex IIA, 7.8.2) .............................172
B.8.4.4.3 Water/sediment studies (OECD Annex IIA, 7.8.3).....

................................................... ........172
B.8.4.4.4 Supplemental studies ...............................

................................................... .........................181
B.8.4.4.5 Summary on fate & behaviour in aquatic systems .....

................................................... .......183

B.8.4.5 Degradation in the saturated zone (OECD Annex IIA, 7.9) .............................................18 8

B.8.5 Impact on water treatment procedures (OECD Annex II IA, 9.6.4)..................................189

B.8.6 Predicted environmental concentration in ground wat er for the active substance

and metabolites (PEC GW 

### ) (OECD Annex AIII, 9.6)........................... .................................190

B.8.7 Predicted environmental concentration in surface wa ter and sediment for the

active substance and metabolites (PEC SW 

### , PEC SED 

### ) (OECD Annex AIII, 9.7, 9.8).........190

B.8.8 Fate and behaviour in the air......................

................................................... ....................191
B.8.8.1 Rate and route of degradation in air (OECD Annex II A, 7.10, IIIA, 9.9)...............................1 91

B.8.9 Predicted environmental concentration in the air (P EC A 

### ) ..............................................193

B.8.10 Definition of the residues (OECD Annex IIA, 7.11)..

................................................... .....194

B.8.11 Monitoring data (OECD Annex IIA, 7.12) .............

................................................... ..........194

B.8.12 Other/special studies (OECD Annex IIA, 7.13, AIII 9 .10) ............................................... ..194

B.8.13 References relied on ...............................

................................................... ........................201

Appendix:

B.8.3 Predicted environmental concentration in soil (PEC S 

### ) for the active substance

and relevant metabolites (OECD Annex IIIA, 9.4, 9.5 ).................................................. ...210

B.8.6 Predicted environmental concentration in groundwate r (PEC GW 

### ) for the active

substance and relevant metabolites (OECD Annex IIIA , 9.6) .........................................214

B.8.7 Predicted environmental concentration in surface wa ter (PEC SW 

### ) and sediment

(PEC SED 

### ) for the active substance and relevant metabolites (OECD Annex IIIA, 9.7, 9.8)..........................................

................................................... ....................................220

B.8.9 Predicted environmental concentration in the air (P EC A 

### ) (OECD Annex IIIA, 9.9).......226

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Abbreviations used in the DAR:

AR applied radioactivity
ASE accelerated solvent extraction BBCH Biologische Bundesanstalt, Bundessortenamt an d Chemical Industry CH crop height DAT days after treatment DFOP double first order in parallel DM dry mass ESI electro spray ionisation EU European Union FOCUS Forum for co-ordination of pesticide fate mod els and their use FOMC first order multi compartment (Gustafson-Holde n) GAP good agricultural practice
HAT hours after treatment HPLC high pressure liquid chromatography HS Hockey-Stick LC liquid chromatography
LOD limit of detection LOQ limit of quantification LSC liquid scintillation counting
MC moisture content MS mass spectroscopy MWHC maximum water holding capacity
NER non-extractable residues NMR nuclear magnetic resonance OD oil dispersion RAD radioactivity detector ROI region of interest RP reversed phase SFO single first order SFO-RB single first order reversible binding TLC thin layer chromatography
TRR total recovered residues
UV ultraviolet WHC water holding capacity

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8 Fate and behaviour in the environment

Spirotetramat is a new systemic broad spectrum insecticide inten ded for the control of sucking pests. Spirotetramat belongs to the chemical class of tetr amic acid derivates (ketoenoles).

Representative formulation The product OD 150 is formulated as an oil dispersion containing 150 g spirotetramat L -1 

### and is used as systemic broad spectrum insecticide against sucking pests. The maximum total application rate in a sea son is 288 g ai ha -1 

### . The intended EU use patterns are described in Tab le B.8-1.

Table B.8-1: Intended uses of spirotetramat as form ulation OD 150.

Culture Use Numberof applications Interval

[d] Application

timing BBCH

code Crop interception [%] Application rate

[kg ai ha -1 ]

Lettuce, head and leafy F a Max. 2 14 BBCH 48 at last application 48 70 0.072

Lettuce, head and leafy G b Max. 2 14 BBCH 48 at last application 48 70 0.072

Citrus, all types F 2 21 BBCH 78 at last application 78 70 0.096 m -1 CH c

, max. 0.288 (max. CH = 3 m)

a F denotes field use
b G denotes glasshouse use

c CH denotes crop height



### Isomerism of the parent substance Two stereo isomers of spirotetramat, cis- and trans -isomer, are possible. The cis-isomer is the active substance, the trans-isomer (AE 1302942) is declare d as an isomeric impurity in a maximum content of 1.5 % (15 g kg -1 

### ). The cis/trans isomers ratio is about 98/2.

Labelling of the parent substance used for environm ental fate studies In environmental fate studies spirotetramat was con ducted using one or two labelling positions (Table B.8-2) as well as the non-labelled parent compound.

Table B.8-2: Labels of spirotetramat used in the en vironmental studies.

Applicants name Abbreviation used in the DAR Structure Mass [g mol -1 ] Chemical name

Azaspirodecenyl-3- 14 C [3- 14 C] NH

O O O O

CH 3

CH 3 H 3 C H O CH 3



### * : #1

Azaspirodecenyl-5- 14 C [5- 14 C] NH

O O O O

CH 3

CH 3 H 3 C H O CH 3



### * : #2 373.45 IUPAC: cis-3-(2,5- dimethylphenyl)-8- methoxy-2-oxo-1- azaspiro[4.5]dec-3-en-4- yl ethyl carbonate

CA index : Carbonic acid, cis-3-(2,5- dimethylphenyl)-8- methoxy-2- oxo-1- azaspiro[4.5]dec-3-en-4- yl ethyl ester

* Indicates labelling

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Other radiolabels were not investigated since the p athway of degradation is sufficiently understood us ing those two radiolabels. When using the [3- 14 

### C] label (benzylic position) the metabolization cou ld be followed until dimethyl-benzoic acid. An oxidative degradati on leading to an elimination of the 14 

### C atom cannot be envisioned without further oxidation of the phenyl system leading to small chain aliphatic compounds j ust containing C, H and O. Such compounds are relativel y simple parts in the natural carbon cycle of soil. Supportive is that high mineralization rates were d escribed for molecules where the respective phenyl ring was labelled instead of the carboxyl group (in : ‘Metabolic Pathways of Agrochemicals’, TR Roberts (Ed.), The Royal Society of Chemistry Information Services , Cambridge, UK, 1998) ). The oxidative degradation of the phenyl ring of aromatic compounds under ring cl eavage by e. g. mono- and dioxygenases leading to degradates entering classical biochemical pathways like the tricarboxylic acid cycle is extensively de scribed in the literature (for a review see Dagley S (1971) Catabolism of aromatic compounds by microorganisms, Adv. Microb. Physiol. 6, 1-46 ); for anaerobic degradation of respective aromatic compounds see Harwood CS, Burchhardt G, Herrmann H, Fuchs G (1999) Anaero bic metabolism of aromatic compounds via the benzoyl-CoA pathway, FEMS Microbiology Reviews 22, 439-458 ). Therefore, it is regarded very unlikely that degradates “beyond” the dimethylbenzoic acid would play any significant role in the environment. It is rather assumed that, as soon as all the labels carrying MA -amide derived degradates are metabolically cut dow n to the respective benzoic acids, these would quickly d egrade to CO 2 

### without the formation of any major phenylspecific degradate in soil.

In the case of the related BCS ketoenol compound Sp iromesifen (Oberon®) a third (phenyl-) label was investigated (in this case concerning the three-met hyl-group-derived mesitylen hydrocarbon). However results showed that no phenyl-label specific metabo lites were found. In the case of the related BCS ke toenol compound Spirodiclofen (Envidor®) the aerobic soil degradation of the related [ 14 

### C-phenyl]-2,4- dichlorobenzoic acid was investigated using four so ils and yielded a mean DT 50 

### value of 9.7 days. From this it is expected that 2,5-dimethylbenzoic acid would be degraded even faster.

Chemical name, structure, molecule mass and occurre nce of metabolites Chemical name, structure, molecule mass and occurre nce of metabolites which were observed in environmental compartments or which were used as re ference substances are given in Table B.8-4.

Table B.8-3: Chemical name, structure, molecule mas s and occurrence of investigated and/or observed metabolites of spirotetramat.

Substance Chemical name Structure Mole

mass [g mol -1 ] Occurrence

(maximum amounts based on mean of replicates)

[% of AR]

Spirotetramatenol CAS: cis-3-(2,5-

Dimethylphenyl)- 4-hydroxy-8- methoxy-1-azaspiro[4.5]dec3-en-2-one 301.39 Aerobic soil: 100.0 (worst case assumption) Aerobic outdoor: 7.8 Anaerobic soil: 54.6 Field soil: na a Soil photolysis: 10.1 Hydrolysis (pH 7): 91.8 Photolysis water: 81.9 Aerobic water/sed.: 79.7 / 41.2

Spirotetramatketohydroxy CAS: cis-3-(2,5-

Dimethylphenyl)- 3-hydroxy-8- methoxy-1- azaspiro[4.5]deca n-2,4-dione 317.39 Aerobic soil: 24.0 Aerobic outdoor: 25.3 Anaerobic soil: 19.3 Field soil: na a Soil photolysis: 20.9 Hydrolysis: - Photolysis water: - Aerobic water/sed.: 17.4 / 42.8

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Substance Chemical name Structure Mole

mass [g mol -1 ] Occurrence

(maximum amounts based on mean of replicates)

[% of AR]

Spirotetramat-MAamide IUPAC: 2 cis-1-

[[(2,5- Dimethylphenyl)hy droxyacetyl]amino ]-4- methoxycyclohexa necarboxylic acid 335.40 Aerobic soil: 5.2 Aerobic outdoor: 6.2 Anaerobic soil: 7.2 Field soil: 4.5 Soil photolysis: - Hydrolysis: - Photolysis water: - Aerobic water/sed.: 7.8 / 2.1

Spirotetramatdi-hydroxy IUPAC: (5s,8s)-3- (2,5- dimethylphenyl)- 3,4-dihydroxy-8- methoxy-1- azaspiro[4.5]deca n-2-one 319.40 Aerobic soil: - Aerobic outdoor: - Anaerobic soil: 3.2 Field soil: - Soil photolysis: - Hydrolysis: - Photolysis water: - Aerobic water/sed.: 4.3 / 5.5

Spirotetramatdesmethyl-enol IUPAC: (5s,8s)-3- (2,5- dimethylphenyl)- 4,8-dihydroxy-1-azaspiro[4.5]dec3-en-2-one 287.36 Aerobic soil: 2.0 Aerobic outdoor: - Anaerobic soil: - Field soil: - Soil photolysis: - Hydrolysis: - Photolysis water: - Aerobic water/sed.: -

Spirotetramatoxo-enol IUPAC: 3-(2,5-

dimethylphenyl)-4- hydroxy-1-azaspiro[4.5]dec3-ene-2,8-dione 285.35 Aerobic soil: 1.1 Aerobic outdoor: - Anaerobic soil: - Field soil: - Soil photolysis: - Hydrolysis: - Photolysis water: - Aerobic water/sed.: 6.0 / 2.3

Dimethyl-benzoic acid IUPAC: 2,5-

Dimethyl-benzoic acid 150.18 Aerobic soil: - Aerobic outdoor: 3.3 Anaerobic soil: - Field soil: - Soil photolysis: 21.8 b Hydrolysis: - Photolysis water: - Aerobic water/sed.: -

Spirotetramatglyoxylic amide IUPAC: (1s,4s)-1- {[(2,5- dimethylphenyl)(o xo)acetyl]amino}- 4- methoxycyclohexa necarboxylic acid 333.39 Aerobic soil: - Aerobic outdoor: 2.3 Anaerobic soil: - Field soil: - Soil photolysis: 4.1 Hydrolysis: - Photolysis water: - Aerobic water/sed.: -

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Substance Chemical name Structure Mole

mass [g mol -1 ] Occurrence

(maximum amounts based on mean of replicates)

[% of AR]

4-Methoxycyclohexanone IUPC: 4-methoxycyclohexanone 128.17 Aerobic soil: - Aerobic outdoor: - Anaerobic soil: - Field soil: - Soil photolysis: 10.0 Hydrolysis: - Photolysis water: 17.5 Aerobic water/sed.: -

4-Methoxycyclohexylaminocarboxylic acid IUPC: (1s,4s)-1-

amino-4- methoxycyclohexa necarboxylic acid 173.21 Aerobic soil: - Aerobic outdoor: - Anaerobic soil: - Field soil: - Soil photolysis: - Hydrolysis: - Photolysis water: 11.3 Aerobic water/sed.: -

Spirotetramatoxo-ketohydroxy IUPAC: 3-(2,5-

dimethylphenyl)-3- hydroxy-1- azaspiro[4,5]deca ne-2,4,8-trione 301.35 Aerobic soil: << 0.5 Aerobic outdoor: - Anaerobic soil: - Field soil: - Soil photolysis: - Hydrolysis: - Photolysis water: - Aerobic water/sed.: -

Spirotetramatphoto-cyclopentyl IUPAC: (1s,4s)-8'- hydroxy-4- methoxy-5'-methyl-2'Hspiro[cyclohexane -1,1'-indeno[1,2- c]pyrrol]-3'(8'H)-

one NH

O O H

O H

299.4 Aerobic soil: - Aerobic outdoor: - Anaerobic soil: - Field soil: - Soil photolysis: - Hydrolysis: - Photolysis water: 42.9 c
Aerobic water/sed.: -

Spirotetramatphoto-2- methylcarbonate IUPAC: ethyl 2- [(5s,8s)-8- methoxy-2-oxo-1-azaspiro[4.5]dec3-en-3-yl]-4- methylbenzyl carbonate O O O

NH

O O H

373.4 Aerobic soil: - Aerobic outdoor: - Anaerobic soil: - Field soil: - Soil photolysis: - Hydrolysis: - Photolysis water: 19.3 c
Aerobic water/sed.: -

Spirotetramatphoto-2-hydroxymethyl IUPAC: (5s,8s)-3- [2- (hydroxymethyl)- 5-methylphenyl]- 8-methoxy-1-azaspiro[4.5]dec3-en-2-one NH

O O H

O H

301.4 Aerobic soil: - Aerobic outdoor: - Anaerobic soil: - Field soil: - Soil photolysis: - Hydrolysis: - Photolysis water: 22.9 c
Aerobic water/sed.: -

Spirotetramatphoto-2-formyl IUPAC: 2-[(5s,8s)-

8-methoxy-2-oxo1-azaspiro[4.5]dec3-yl]-4- methylbenzaldehy de NH

O O H

O

299.4 Aerobic soil: - Aerobic outdoor: - Anaerobic soil: - Field soil: - Soil photolysis: - Hydrolysis: - Photolysis water: 11.5 c
Aerobic water/sed.: -

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Substance Chemical name Structure Mole

mass [g mol -1 ] Occurrence

(maximum amounts based on mean of replicates)

[% of AR]

Spirotetramatenol-dimer 1 d Not available 600 Aerobic soil: 5.0 Aerobic outdoor: 1.5 Anaerobic soil: 7.0 Field soil: < 0.2 Soil photolysis: 2.3 Hydrolysis: - Photolysis water: - Aerobic water/sed.: -

Spirotetramatenol-dimer 2 d Not available 600 Aerobic soil: 3.6 Aerobic outdoor: 0.9 Anaerobic soil: 4.6 Field soil: < 0.2 Photolysis water: - Soil photolysis: - Hydrolysis: - Aerobic water/sed.: -

a Not applicable owing to distinct storage instabili ty of spirotetramat-enol (degrading to spirotetrama t-ketohydroxy during frozen storage)
b In an outdoor metabolism study, conducted with the formulation on bare soil including natural sunligh t, dimethyl-benzoic acid was shown not to exceed 3.3 % of AR. Not considered rel evant for environmental risk assessment.

c Considered as photo-rearrangement products owing t o the highly artificial experimental conditions (st erile buffer solution). Not considered to occur under environmental conditions (as demonstrated in an additional photolysis study conducted in sterile natural water). Not considered relevant for environmental r isk assessment.
d Considered as artefacts owing to hot-spot applicat ion in the laboratory, formation under environmenta l conditions (formulated ai) negligible. Not considered relevant for environment al risk assessment.



### Table B.8-4 gives an overview on metabolites of spi rotetramat observed in environmental compartments
which are considered relevant for further risk asse ssment (EU risk assessment).

Table B.8-4: Summary of metabolites [maximum occurr ence in % of AR] of spirotetramat considered relevant for a further risk assessment owing to tri gger exceedance (EU risk assessment).

Compartment Soil Soil with regard to groundwater Surface
water Sediment

Trigger > 10 % > 10 % or 2 consecutive samples > 5 %

or steady increase > 10 % > 10 % Remarks

Spirotetramat-enol 100.0 a 100.0 a 79.7 41.2 Spirotetramat-ketohydroxy 24.0 b 24.0 b 17.4 42.8 Spirotetramat-MA-amide < b 5.2 b < < 4-Methoxy-cyclohexanone 10.0 - - - Photolysis produc t4-Methoxy-cyclohexylaminocarboxylic acid - - 11.3 - Photolysis product

< denotes ‘observed below the trigger’ - denotes ‘not observed’

a Worst-case assumption
b Data derived from the spirotetramat-enol laborator y degradation study

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1 Route and rate of degradation in soil

B.8.1.1 Route of degradation (OECD Annex IIA, 7.1)

B.8.1.1.1 Aerobic degradation in soil (OECD Annex I IA, 7.1.1)

The route of aerobic degradation (metabolism) of sp irotetramat in soils was established in one laborat ory study (which was also used for degradation rate cal culations) and one supplemental outdoor study:

• Aerobic degradation/metabolism in 4 soils (20 °C, [3- 14 

### C] label)

• Aerobic degradation under outdoor conditions in 2 soils ([3

- 14 

### C] label)

Further, a metabolism/degradation study was conduct ed with spirotetramat-enol, which is the major metabolite of spirotetramat. This study was perform ed since the parent study was not considered valid for quantitative distribution pattern of the metabolite s owing to analytical reasons.

• Aerobic degradation/metabolism of spirotetramat-enol in 4 soils (20 °C, [3- 14 

### C] and [5- 14 

### C] label)

Reference: Aerobic degradation/metabolism of BYI8330 in four d ifferent soils Author(s), year: Babczinski, P., 2005 Study/report number: M-256849-02-2, MEF-04/169, ame nded 05.05.2006 Guideline(s): US EPA, Subdivision N, Section 162-1 (1982); OECD 307 (2002); 95/36/EC amending 91/414/EEC; Japanese MAFF, 12 Nohsan 8147 (2000, 2001)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: [Azaspirodecenyl-3- 14 

### C]-spirotetramat (sample ID: BECH 0755); specific radioactivity 3.71 MBq mg -1 

### (100.2 μCi mg -1 

### ); Radiochemical purity: > 99 % (acc. radio-HPLC and -TLC); chemical purity: > 99 % (HPLC , UV detection at 210 nm); identity and purity of the stock solution was check ed Reference substances: Spirotetramat (unlabelled), s pirotetramat-enol, spirotetramat-ketohydroxy, spirotetramat-MA-amide, spirotetramat-desmethyl-eno l, spirotetramat-oxo-enol, mandelic acid, dimethyl-benzoic acid, spirotetramat -enol-dimer 1 Test system: Static incubation test system Acclimatisation time: 20 – 39 days (application 37 - 42 days after sampling) Test conditions: Aerobic conditions; for ‘Molino’ s oil (US soil, metabolism soil) replicates were set up, for EU soils (kinetic soils) all sampling dates except of DAT-0 were analyzed via single test flasks Moisture adjusted soils (50 % MWHC [EU soils] or 75 % of 1/3 bar moisture [US soil]) maintained at 20 ± 1 °C in the dark. At DAT- 0 the absolute microbial biomass carbon contents corresponded to 6.4, 3.1, 2 .6 and 1.9 % relative to the soil organic carbon content of the soils ‘Hoefchen’ , ‘Laacherhof AXXa’, ‘Laacherhof AIII’ and ‘Molino’, respectively Test duration: EU soils – 50 days; the EU soils exp eriments were terminated earlier as after one
or two weeks at least 90 % of the test item was tra nsformed and at least > 5 %
CO 2 

### was formed (according OECD 307) US soil – 360 days (according EPA § 162-1) Sampling intervals: EU soils – 0 (duplicate sample) , 0.25, 1, 3, 7, 14, 30 and 50 days (single samples ) US soil – 0, 0.25, 1, 3, 7, 14, 30, 50, 86, 126, 17 9, 270 and 360 days (duplicate

### – 11 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

samples) Application: EU soils – 0.128 mg kg -1 

### (nominal) dry soil, US soil – 0.768 mg kg -1 

### (nominal) dry soil, equivalent to 0.288 kg ai ha -1 

### (calculation based on homogeneous distribution for 2.5 cm soil depth [EU soils] and 15 cm soil dep th [US soil] and 1.5 g cm -3 

### soil density); supplied test item was dissolved in 2 mL acetonitrile A so-called “hot spot application” (soil spiking by a pipette with a low volume of higher concentrated organic test solution) was used as the test item is highly degradable and has a low water solubility Analyses: Ambient extraction procedure : 100 g dry mass (DM) soil samples were extracted 3 times with 80 mL of acetonitrile/water (1:1, v/v, c ontaining 0.5 % formic acid) Aggressive extraction procedure : To obtain information about the bound residues the residual soil already extracted using the ambie nt procedure was additionally extracted twice with 80 mL of acetonitrile/1N hydro chloric acid (1:1, v/v, pH ca. 1). Finally the samples were extracted with pure aceton itrile (1 x 50 mL) using the same technique. The spirotetramat residues were radio-assayed by LS C and analyzed separately in the combined ambient and combined aggravated ext racts by HPLC on reversed phase (C-18) columns. Solid samples (i.e. soil and paper filters) were combusted
and 14 

### C levels were measured using LSC. In order to ident ify parent compound and the transformation products co-chromatography a nd spectrometric methods
were used. Volatile traps: Organic volatiles: polyurethane foa m, CO 2 

### : soda lime, CO 2 

### identity confirmed by
BaCO 3 

### precipitation Analytical techniques: LSC, RP-HPLC, Si-60 silica T LC, LC-MS/MS, NMR
Detection limits: For LSC: LOQ ≤ 0.2 % of AR, for CO 2 

### samples LOQ < 0.1 % of AR;
for HPLC: LOQ ≤ 0.2 % of AR Kinetic evaluation: Simple first order (SFO) kineti cs; ModelManager, vers. 1.1., C 0 

### calculated by the model; only parent modelled

Table B.8.1.1.1-1: Main soil properties.

Parameter ‘Molino’ ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’

Source Florida, USA Monheim, Germany Monheim,

Germany Burscheid,

Germany

Texture (USDA) Tifton fine sandy loam Sandy loam Silt loam Silt

Soil taxonomic classification (USDA) Fine-loamy, kaolinitic, thermic Plinthic Kandiudults Sandy, mixed,

mesic Typic Cambudolls Loamy, mixed,

mesic Typic Argudalfs Loamy, mixed,

mesic Typic Argudalfs

SCS soil series Ultisol Mollisols, Udolls Alfisols, Udalfs Alfisols, Udalfs

Sand (USDA) [%] 77.3 72.4 36.9 8.5 Silt (USDA) [%] 12.7 22.6 51.1 81.3 Clay (USDA) [%] 10.0 5.0 12.0 10.2 pH (H 2 O) 6.1 7.2 7.4 7.6 pH (CaCl 2 ) 5.4 6.3 6.5 6.7 pH (KCl) nd 6.4 nd 7.0 Organic matter [%] 1.60 1.75 1.43 3.63 Organic carbon a [%] 0.93 1.02 0.83 2.11 Carbonate (CaCO 3 ) [%] < 0.1 < 0.1 < 0.1 0.9 CEC [meq/100 g DM] 6 8 8 15 WHC at pF2.5 (FC at 0.33 bar) [g/100 g DM] 18.4 29. 1 20.7 35.6

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Parameter ‘Molino’ ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’

MWHC [g/100 g DM] Not available 34.4 36.4 63.1
Microbial C b [mg microbial C/kg soil DM] start 180 312 213 1341
Microbial C b [mg microbial C/kg soil DM] DAT-59 118 261 202 730
Microbial C b [mg microbial C/kg soil DM] DAT-360 77 - - -

nd denotes not determined

a Organic C = organic matter / 1.724
b Substrate induced respiration



### Since preliminary experiments with a so-called ‘bul k soil application’ showed low DAT-0 recoveries and the test item and/or its degradate(s) quickly degraded and firmly bound to the soil, for the present study a socalled ‘hot-spot’ application was used. Due to this application form an immediate DAT-0 extraction was possible with high efficiency (93.1 to 99.6 %).

Under the used acidic extraction solvents the test item showed good stability values whereas the major soil degradate spirotetramat-enol showed an instability under these processing conditions. Using slightly a lkaline conditions, enol degradation could be largely preve nted (additional experiment, Figure B.8.1.1.1-1). I n the absence of soil, spirotetramat-enol was stable unde r both the acidic and alkaline processing condition s. However, alkaline processing conditions would be de trimental for the stability of the parent compound. Therefore, within this parent degradation study it was not possible to use a processing procedure guaranteeing for the stability of the parent and its major degradation product. To overcome these d ifficulties, an additional aerobic degradation study using an al kaline extraction procedure for spirotetramat-enol was conducted (see separate study, Babczinski, 2006b).

Figure B.8.1.1.1-1: Additional experiment: 0-DAT recovery of applied spirotetramat-enol using acidic or slightly alkaline extraction.

Findings: All values are given as mean values of the replicat es.

Total mass balance was in the range of 85.7 % to 10 0.5 % of AR whereas only two out of 37 mass balance values were below 90 % (85.7 and 86.8 %).

At the end of the incubation time the amount of 14 

### CO 2 

### reached 15.3 % of AR in soil ‘Molino’ (DAT-360) an d 12.2 to 19.4 % of AR in the EU soils (DAT-50). The highest CO 2 

### formation rate was formed in the soil ‘Hoefchen’ expected due to its highest content of m icrobial biomass. Compared to the other soils, soil
‘Molino’ showed at DAT-50 the lowest mineralization rate. No other volatile compounds were detected.

### – 13 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

The portion of non-extractable radioactivity ( NER ) rapidly increased reaching its peaks at levels of 21.3 % of AR to 35.2 % of AR after one to three days after application. At the termination of the studies (US soil: DAT-360, EU soils: DAT-50) bound residues reached a level of 21.5 % of AR to 31.0 % of AR. The highest value was measured in ‘Molino’ soil (mean value 35. 2 % of AR, DAT-3). A partitioning of the heterogene ous NER into humic acid, fulvic acid and humin like fra ctions was observed, with a major portion belonging to the fulvic acid fraction and with a significant portion bound to soil in the insoluble humin fraction.

Under standard conditions (20 ± 1 °C, 50 % MWHC [EU soils] or 75 % of 1/3 bar moisture [US soil]) a ra pid decline of spirotetramat was observed in all four s oils, dropping below 10 % of AR after one day. It w as shown that the degradation of spirotetramat correla tes with the soil biomass content indicating that t he degradation is, at least in part, microbial. The fa st decline of the parent was accompanied in all soi ls by a rapid appearance of the major degradates spirotetramat-enol (max. 24.3 % of AR, DAT-3) and spirotetramat-ketohydroxy (max. 16.3 % of AR, DAT-1). Beside of these two ma in metabolites two spirotetramat-enol-dimers ( spirotetramat-enol-dimer 1 : max. 12.7 % of AR, DAT-7; spirotetramat-enoldimer 2 : max. 9.2 % of AR, DAT-30) and spirotetramat-MA-amide (max. 6.4 % of AR, DAT-179) were identified. Two minor metabolites were spirotetrama t-desmethyl-enol (max. 3.7 % of AR, DAT-30/179) and spirotetramat-oxo-enol (max. 1.2 % of AR, DAT-50). Further, the large number of minor peaks (not ident ified) were summed up under unidentified radioactivity (ma x. single peak < 5 % of AR).

It was not possible to calculate valid DT 50 

### values for the soil degradates due to plateauing t owards study termination in several cases. This effect was expla ined by several reasons: influence of the microbiol ogical activity due to the application solvent, the “hot-s pot” application has caused ‘artifactual’ formation of some transformation products (enol-dimers), influence of the level of spirotetramat-enol and other degradat es due to reformation of spirotetramat-enol from spirotetr amat-enol-dimers and due to the chosen processing procedure (acidic extraction procedure) which is ex pected as partly destructive for the spirotetramat- enol, respectively. For these reasons, separate studies u sing spirotetramat-enol as test item were performed . The
metabolite kinetics is not assessed in detail withi n this study. The calculated DT 50 

### values of spirotetramat are
listed in Table B.8.1.1.1-6.

Dimeric spirotetramat-enol soil “degradates”:
The dimerization issue is discussed in detail under point B.8.11 (position paper Hellpointner et al., 2007).

Metabolic pathway: Spirotetramat was metabolized via three degradation pathways which all started with the hydrolytic cle avage of the spirotetramat ester into the spirotetramat-e nol. In the major degradation pathway the spirotetr amatenol will be oxidized at the benzylic carbon positi on into spirotetramat-ketohydroxy which is hydrolyt ically opened into the spirotetramat-MA-amide and finally mineralized into CO 2 

### . In the second (minor) pathway, demethylation of the spirotetramat-enol into spirot etramat-desmethyl-enol and oxidation of this degrad ate into the spirotetramat-oxo-enol occurred. In a thir d degradation pathway, the spirotetramat-enol-dimer 1 and spirotetramat-enol-dimer 2 were pretended to be bui lt through oxidative enol dimerization, but this pa thway is regarded as non relevant under environmental condit ions (for more detailed information see point B.8.1 1, position paper Hellpointner et al., 2007).

### – 14 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.1.1-2: Distribution and recovery of app lied radioactivity [% of AR] after application of [3- 14 

### C] labelled spirotetramat to 4 soils (mean of dupli cate samples in ‘Molino’ soil).

Extractable Soil DAT Organic/

aqueous extract Aggravated
extract Total CO 2 NER Total

0 94.2 3.5 97.7 nd 0.4 98.1
0.25 68.1 12.2 80.3 0.5 19.7 100.5

1 40.1 22.8 63.0 1.7 31.8 96.5
3 30.0 25.7 55.7 3.7 35.2 94.6

7 30.4 25.8 56.2 5.9 33.0 95.1

14 27.1 24.8 51.9 7.6 30.5 90.1
30 33.1 24.3 57.5 8.4 31.5 97.4
50 31.1 24.6 55.6 9.7 30.3 95.6
86 30.6 24.3 54.9 15.5 29.9 100.4
126 31.9 26.6 58.5 12.1 27.6 98.2
179 28.9 24.0 52.8 15.4 27.8 96.0
270 29.5 25.7 55.1 14.8 27.2 97.1 ‘Molino’

360 26.0 25.9 51.8 15.3 27.0 94.2

0 a 93.1 2.9 96.0 nd 0.1 96.2
0.25 62.1 8.0 70.1 0.3 16.4 86.8

1 53.8 11.9 65.7 1.8 27.2 94.8
3 55.7 12.0 67.7 3.1 25.4 96.1

7 50.0 12.4 62.3 6.3 26.2 94.8

14 43.6 9.5 53.1 8.3 24.3 85.7
30 42.5 12.1 54.6 10.0 27.9 92.5 ‘Laacherhof AXXa’

50 44.2 12.7 56.9 12.2 25.5 94.7

0 a 91.8 2.6 94.4 nd 0.1 94.5
0.25 72.8 8.2 81.1 0.2 14.1 95.4

1 61.1 10.9 71.9 1.5 21.3 94.7
3 60.7 12.5 73.2 3.0 21.0 97.1

7 63.6 10.9 74.6 2.1 19.0 95.7

14 57.8 11.1 68.9 6.9 17.4 93.2
30 55.0 11.3 66.3 10.7 20.6 97.6 ‘Laacherhof AIII’

50 49.1 13.6 62.7 15.4 21.5 99.6

0 a 94.8 3.5 98.2 nd 0.3 98.5
0.25 49.6 16.3 65.8 0.3 32.3 98.4

1 48.7 15.1 63.8 1.3 30.9 96.0
3 39.2 18.7 57.9 3.2 34.5 95.6

7 41.4 19.9 61.3 4.8 32.8 99.0

14 27.7 18.3 46.1 11.4 33.8 91.2
30 30.3 18.7 49.1 13.3 34.6 97.0 ‘Hoefchen’

50 28.4 18.8 47.2 19.4 31.0 97.6

nd denotes not determined

a Mean of duplicate samples



### Table B.8.1.1.1-3: Characterization [% of AR] of no n-extractable soil residues (NER). Sampling DAT-3 DAT-7 DAT-30 DAT-50 a

Soil ‘Hoefchen’ ‘Molino’ ‘Laacherhof AXXa’ ‘Laacherhof AIII’

Total NER 33.8 32.4 27.0 20.9
Fraction humic acids 11.2 13.5 9.7 3.9
Fraction fulvic acids 16.0 16.7 12.9 12.8
Fraction humin substances 8.6 5.6 5.0 4.2 Processing balance [%] 105.7 110.2 102.1 100.1

a ‘DAT-60’ stated in the original report probably a typo– 15 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.1.1-4: Characterization of extractableradioactivity [% of AR] after application of [3-14C] labelled spirotetramat to ‘Molino’ soil (individualreplicates).SpirotetramatSpirotetramat-desmethyl-enolSpirotetramat-oxo-enolSpirotetramat-enolSpirotetramat-enol-dimer 1Spirotetramat-enol-dimer 2Spirotetramat-ketohydroxySpirotetramat-MA-AmideUnidentifiedaSoil DATA B A B A B A B A B A B A B A B A B093.5 98.6 nd nd nd nd nd nd nd nd nd nd nd nd nd nd2.3 1.00.2556.2 48.1 nd nd nd nd 4.4 5.6 nd nd nd nd6.69.4 0.3 0.7 14.2 15.1115.0 15.7 0.9 1.2 nd nd8.89.6 0.6 0.5 1.3 1.67.59.8 2.1 2.8 27.1 21.438.1 5.9 0.9 1.1 nd 0.311.115.41.1 0.3 0.5 0.610.613.5 1.7 2.2 22.7 15.577.2 5.8 3.8 2.4 nd 1.010.310.7 1.7 1.6 0.4 0.412.810.8 4.1 4.3 16.7 18.4144.5 4.5 2.1 1.3 0.6 0.99.47.83.62.0 2.2 0.97.67.9 3.5 4.9 21.2 19.0305.8 4.3 2.94.5nd nd5.27.2 3.4 2.8 2.1 1.59.49.35.6 5.3 21.9 23.7503.9 4.2 3.1 2.0 nd1.28.26.6 3.5 2.1 2.8 1.010.89.05.2 4.7 19.0 24.0863.7 3.7 4.3 2.3 nd nd7.0 4.0 1.6 1.53.21.312.68.65.15.7 18.3 26.71263.3 2.6 3.1 3.3 nd nd6.58.2 3.0 1.3 1.4 0.612.715.75.46.6 21.2 22.21793.7 3.3 3.8 3.7 0.8 nd 4.9 4.2 2.0 1.6 1.7 0.913.58.66.66.2 17.4 22.72702.4 4.0 3.5 2.7 nd nd 3.6 4.9 1.2 0.9 1.6 nd11.214.25.57.025.8 21.7‘Molino’3603.5 3.5 0.9 2.0 nd nd 2.5 2.9 0.5 0.4 0.9 0.713.713.57.05.4 24.1 22.2nd denotes not determinedaUp to 18 HPLC peaks/DAT; max. value for a single peak: 4.0 % of AR– 16 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.1.1-5: Characterization of extractableradioactivity [% of AR] after application of [3-14C] labelled spirotetramat to ‘Laacherhof AXXa’,‘Laacherhof AIII’ and ‘Hoefchen’ soil (HPLC, no replicates available).Soil DAT SpirotetramatSpirotetramatdesmethylenolSpirotetramat-oxo-enolSpirotetramat-enolSpirotetramat-enol-dimer 1Spirotetramat-enol-dimer 2Spirotetramat-ketohydroxySpirotetramat-MA-amideUnidentifieda0b91.8 nd nd nd nd nd nd nd 4.30.2538.3 0.2 nd6.8 2.3 3.110.6 0.4 8.418.3 1.9 nd18.8 2.46.216.32.3 9.733.1 1.51.019.03.55.014.8 3.0 16.973.52.7nd11.97.0 2.712.86.015.8142.6 1.6 0.410.27.05.77.4 4.4 13.7302.2 2.6 nd8.87.45.55.1 3.5 19.5‘LaacherhofAXXa’502.5 1.8 nd9.57.56.95.9 3.1 19.60b91.6 nd nd nd nd nd nd nd 2.80.2541.2 nd nd9.4 4.4 4.210.5 0.2 11.118.9 0.4 nd20.8 3.37.215.01.9 14.333.4 nd nd24.33.56.314.3 3.1 18.273.52.7nd15.112.74.212.95.418.1142.8 1.9 nd15.59.48.98.4 3.9 18.2303.0 2.1 nd11.49.89.26.6 3.3 20.8‘LaacherhofAIII’503.4 2.3 nd9.97.78.15.6 3.3 22.30b93.2 nd nd nd nd nd nd nd 50.2511.6 1.0 0.611.6 4.3 3.714.72.4 1615.8 0.6 nd17.2 2.1 3.89.4 2.8 22.231.9 1.7 nd16.92.5 2.111.8 4.8 16.073.72.3 1.114.15.71.611.05.316.4142.02.30.48.4 4.2 3.5 4.3 2.8 18.2302.1 1.6 nd9.85.5 3.8 3.8 2.0 20.5‘Hoefchen’502.6 1.6 nd10.4 4.96.02.4 1.1 18.3nd denotes not determinedaTotal % of AR for unidentified discrete componentsand streaking, none exceeding 5 % of AR individuallybMean of two replicates

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.1.1-6: Calculated DT 50 

### and DT 90 

### values [days] of [3- 14 

### C] labelled spirotetramat in 4 soils based on SFO kinetics.

Soil DT 50 [days] DT 90 [days] R 2 Kinetics

‘Molino’ 0.327 1.090 0.974 SFO
‘Laacherhof AXXa’ 0.210 0.697 0.991 SFO
‘Laacherhof AIII’ 0.232 0.770 0.989 SFO
‘Hoefchen’ 0.083 0.277 0.991 SFO



### Conclusion: Spirotetramat was shown to rapidly biodegrade in al l four tested soils (1 US soil and 3 EU soils) unde r standard aerobic conditions, applied at a concentra tion equivalent to 0.288 kg ai ha -1 

### . Dissipation followed
SFO kinetics with calculated half-lives of 0.08 – 0 .33 days at 20 °C. The evolved 14 

### CO 2 

### (no volatile organics) was about 12.2 – 19.4 % of AR at study termination (DAT-50, EU soils) whereas soil ‘Hoefchen’ showed t he highest mineralization rate pretended caused by its highest content of microbial biomass. The US soil accounted for 9.7 % of AR (DAT-50) and increased to 15.3 % of AR at study termination (DAT-360). Besid es

14 

### CO 2 

### a portion of the applied radioactivity was found to be integrated into NER (max. 35.2 % of AR in soil ‘Molino’, mean value/DAT-3). This portion of radioa ctivity timely occurs very fast after application.
Fractionation demonstrated the NER residue to be of heterogeneous nature with a major portion belongin g to the fulvic acid fraction. The metabolites were i dentified as spirotetramat-enol (max. 24.3 % of AR, mean value, DAT-3), spirotetramat-ketohydroxy (max. 16.3 % of AR, mean value, DAT-1), spirotetramat-enoldimer 1 (max. 12.7 % of AR, mean value, DAT-7), spirotetramat-enol-dimer 2 (max. 9.2 % of AR, mean value, DAT-30), spirotetramat-MA-amide (max. 6.4 % of AR, mean value of replicates, DAT-1 79), spirotetramat-desmethyl-enol (max. 3.7 % of AR, mea n value, DAT-179) and spirotetramat-oxo-enol (max. 1.2 % of AR, mean value, DAT-7).

In the context of the relevance as soil ‘major’ met abolites (> 10 % of AR), spirotetramat-enol and spirotetramat-ketohydroxy should be taken into acco unt. With regard to the groundwater also spirotetra matMA-amide has to be considered further (following th e trigger for the EU risk assessment). The spirotet ramatenol-dimers are regarded as artifactual degradates (for details refer to B.8.11, position paper Hellpo intner et al., 2007) and, therefore, both are not relevant fo r further consideration.

It was assumed that the present study gives just va lid data related to the disappearance of spirotetra mat, but not the true quantitative distribution pattern of t he metabolites resulting from the rapidly formed sp irotetramatenol. Consequently, the kinetic evaluation was base d on the spirotetramat-enol study (Babczinski, 2006 b), which avoids the existing enol-stability problems ( acidic extraction conditions needed for spirotetram at) in the parent study. Further, the artificial hot spot/orga nic solvent application needed in the parent study leading to artificially high dimer amounts were regarded as un certainty factors in the metabolic profiling (see B .8.11, position paper Hellpointner et al., 2007).

A second label study was considered unnecessary sin ce significant mineralization was observed and no metabolite indicated molecule breakage at significa nt amounts.

Reviewer comments:

• More detailed information on the dimerization issu e is summarized under point B.8.11 (Position paper Hellpointner et al., 2007).

• Degradation kinetics of the parent was re-calculat ed in a separate study (Ramanarayanan and Röpke, 2006).

• Owing to the observed instability of spirotetramat -enol under the acidic extraction used in this stud y, the additionally conducted spirotetramat-enol degra dation study (Babczinski, 2006b), which avoids these problems, is considered the only reliable bas is for the determination of maximum occurrence

### – 18 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

and kinetic behaviour of spirotetramat-enol and the subsequent metabolites (spirotetramatketohydroxy and spirotetramat-MA-amide) in aerobic soils degradation.

• On request the notifier (BCS) provided further inf ormation in respect to the
identification/characterization of the NER fraction : BCS: The studies were conducted according to establ ished guidelines. Harsh extraction methods were employed to extract as much of NER fro m the soil as possible. The aggressive method used in the current study (2 times ACN/1 N H Cl, pH 1) solubilized a lot of the soil matrix (dark brown extracts). Further strong alkaline extr action (0.5 M NaOH) was conducted for BR
characterization as humic and fulvic substances. Th ese treatments are considered as adequate
for BR identification/ characterization in the sens e of citations [1] to [5] [see below] . Any new methods to characterize NER (or BR) would not add m uch value to the risk assessment because NER is immobile and not readily bioavailabl e or bioaccessable. [1] FOCUS (2006) “Guidance document on estimating pe rsistence and degradation kinetics from environmental fate studies on pesticides in EU regis tration” Report of the FOCUS Work Group on Degradation Kinetics, EC Document reference Sanco/1005 8/2005 version 2.0 [2] SETAC (1995) Procedures for assessing the environme ntal fate and ecotoxicity of pesticides, ed. M R Lynch [3] Pesticide Reregistration Rejection Rate Analysis Environmental Fate, US EPA, EPA 738-R-93-0110,

1993 [4] ECPA Position Paper on Soil Non-Extractable Residues, Brussels, March 3, 2000, D/00/SuM/5277 [5] SCP Opinion on Draft GD on persistence in soil (D G IY-9188/IV/97-Rev. 5, 1998) dated 1999

### – 19 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: Outdoor Metabolism of [Azaspirodecenyl-3- 14 

### C]BYI08330 in two soils Author(s), year: Babczinski, P., 2006a Study/report number: M-270597-01-2, MEF-06/041 Guideline(s): Supportive non-guideline study, but a pproached to US EPA, Subdivision N, Section 162-1 (1982); OECD 307 (2002); EC-Directiv e 91/414/EEC Annex I Part 7 and Annex II Part 9; SETAC Procedures for Assessi ng the Environmental Fate and Ecotoxicity of Pesticides.
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: [Azaspirodecenyl-3- 14 

### C]-spirotetramat (sample ID: BECH 1569); specific radioactivity 3.67 MBq mg -1 

### (99.1 mCi mg -1 

### ); radiochemical purity: > 99 % (acc. radio-HPLC) and > 98 % (acc. radio -TLC); identity and purity of the stock solution
was checked Reference substances: Spirotetramat (unlabelled), s pirotetramat-enol, spirotetramat-ketohydroxy, spirotetramat-MA-amide, spirotetramat-glyoxylic-ami de, dimethyl-benzoic acid, methoxycyclohexanone, glyoxylic amide, spirotetrama t-enol-dimer 1, spirotetramat-enol-dimer 2 Test system: Two semi-disturbed (without sieving) s oils (‘Laacherhof AIII’ [EU soil] and ‘Molino’ [US soil]) were poured into a planting container ap prox. 1 m 2 

### area which was divided into two halves by a separating plate. Soil s were placed for 127 days under outdoor climatic conditions (open system, pro tected for rainfall, exposed to natural temperature, humidity and light-dark cycles ). The bottom was open for drainage. Non-balance-study since no volatiles and residues other than those in the treated soil were addressed; the soils were kep t bare and free of weeds, and irrigation was performed for adequate moisture. Test duration: 127 days (sampling intervals: 1, 7, 14, 28, 63, 127 days (207 days – for TRR determination only) Application: [Azaspirodecenyl-3- 14 

### C]-spirotetramat formulated as an OD 100 was applie d at 94.6 % of the highest recommended single use rate f or field application (288 g/ha); application was carried out using a computer controlled track sprayer fitted with a flat fan nozzle. Application date: 2004-06-2 8
Weather data:

Sampling: Four soil cores (5 cm in diameter, 20 cm in depth) were randomly taken from each half of the soil container at each sampling date. T he upper 10 cm of the sampled soil cores were air-dried at ambient temperature (o vernight), homogenized and their dry weight determined. Then a tenth of the we ight of each core was taken, combined and mixed, and the mixed sample used for e xtraction. The remaining sample was used for TRR determination by combustion /LSC. Analyses: Soil samples were extracted three times w ith acidic acetonitrile/water/formic acid (1000/1000/1 each 80 mL, conventional method), foll owed by twice acetonitrile/1N HCl (1/1; each 80 mL, aggressive method). Conventio nal and aggressive extracts

### – 20 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

were combined, radio-assayed by LSC and analyzed se parately in the combined ambient and combined aggravated extracts by HPLC. F rom DAT-28 on, the profiling HPLC method was changed (original HPLC me thod 2 [Kromasil] was replaced by HPLC method 1 [Purospher]). Therefore, all extracts from DAT-1, DAT-7 and DAT-14 were either re-analyzed (DAT-7) or newly processed after storage of the original extract in a freezer (DAT-1 , DAT-14), and the spirotetramat residues were analyzed by HPLC. Radio-TLC was used for confirmatory identification of isolated metabolites only (i.e. H PLC cuts spiked with nonradiolabelled referenced substances). Identificatio n of the transformation products was performed by co-chromatography with reference s ubstances and LC/MS spectrometry. Since the study was conducted under o utdoor conditions, collection
of 14 

### CO 2 

### and volatile organic compounds was not possible, a nd therefore, the total recovered radioactivity was lower than the total ap plied radioactivity (100 % of AR). Solid samples (i.e. soil and paper filters) we re combusted and 14 

### C levels were measured using LSC. Detection limits: LSC: LOQ < 0.1% of AR, HPLC: LOQ ≤ 0.3 % of AR. Analytical techniques: LSC, RP-HPLC, Si-60 silica T LC, LC-MS, LC-MS/MS, NMR Kinetic evaluation: Single first order (SFO) kineti cs; ModelManager, vers. 1.1., C 0 

### was set to 100 %

Table B.8.1.1.1-7: Main soil properties.

Parameter ‘Laacherhof AIIIa’ ‘Molino’

Location Germany USA, Florida Texture (USDA) Loam Sandy loam

Soil taxonomic classification (USDA) Loamy, mixed, mesic Typic Argudalfs Fine-loamy, kaolinitic, thermic Plinthic Kandiudults
SCS soil series Alfisols, Udalfs Ultisol Sand (USDA) [%] 44.7 77.9 Silt (USDA) [%] 38.3 13.6 Clay (USDA) [%] 17.1 8.6 pH (in water) 7.4 6.0 pH (in CaCl 2 ) 6.7 5.4 pH (in KCl) 6.7 5.4 Organic matter [%] 1.7 1.4 Organic carbon a [%] 1.0 0.8 Carbonate (CaCO 3 ) [%] < 0.1 < 0.1 CEC [meq/100 g DM] 9.3 4.2 MWHC [g/100 g DM] 45.6 22.6
Soil biomass b [mg microbial C/kg soil DM] DAT-2 349 99
Soil biomass b [mg microbial C/kg soil DM] DAT-127 301 55

a Organic C = organic matter / 1.724

b Substrate induced respiration



### Findings: The total radioactive residues (TRR) ranged from 73 .8 to 24.9 % of AR for loam soil ‘Laacherhof AIIIa’ at DAT-1 and DAT-127, respectively. The corresponding values for soil ‘Molino’ were 94.6 to 22.8 % of AR after the two respective sampling intervals. Radioactive losses by mineralization, volatilization or translo cation were not determined since the study was conducted u nder outdoor conditions in an “open system”. No drainage water was obtained during the study. From the results of other degradation studies (e.g. Babczinski, P., 2005) it can be concluded that a ma in dissipation path will be via mineralization to 14 

### CO 2 

### rather than volatilization or leaching (Babczinski, P., 2006c). Two inconsistent TRR values (at DAT-63 [soil ‘Laacherhof AIIIa’] and DAT-127 [soil ‘Molino’]) we re obtained probably caused by a local inhomogenity during application. Therefore, additional TRR deter mination was performed late after the termination o f the

### – 21 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

regular study (DAT-127), at DAT 207. These values w ere mentioned as outliers and were therefore correc ted by interpolation. The corrected values were used fo r kinetic evaluation.

Non-extractable residues increased up to 13.2 % and 16.8 % of AR in the two soils at DAT-28 and DAT-63

, respectively, and then decreased towards the end of the study to 8.5 % and 5.4 % of AR in the two soil s, respectively. No 14 

### CO 2 

### were measured since the outdoor study design did n ot allow for analysis of volatile radioactivity.

Kinetics analyses showed that spirotetramat degrade d rapidly with a DT 50 

### of 1.2 and 2.9 days in soils ‘Laacherhof AIIIa’ and ‘Molino’, respectively (mean 2.1 days). The calculated DT 90 

### values were about 4.1 and 9.6 days (mean 6.9 days), respectively. During the course of the study a large number of degradates wa s observed in both soils (at maximum 39 HPLC regions of interest [ROIs] were set at a single sampling interval). The total unidentified radioactivity con sisted on maximum of 14 and 29 HPLC ROIs and diffus e radioactivity per sampling interval for soils ‘Laac herhof AIIIa’ and ‘Molino’, respectively.

As major degradate spirotetramat-ketohydroxy (max. 25.3 % of AR, DAT-14) was detected. Spirotetramatenol was detected with max. 7.8 % of AR (DAT-7). As minor degradates were identified: spirotetramat-MA

amide (max. 6.2 % of AR, DAT-28), dimethyl-benzoic acid (max. 3.3 % of AR, DAT-7), spirotetramatglyoxylic amide (max. 2.3 % of AR, DAT-7), spirotet ramat-enol-dimer 1 (max. 1.5 % of AR, DAT-28), spirotetramat-enol-dimer 2 (max. 0.9 %, DAT-7) and spirotetramat-ketohydroxy-carboxy (max. 1.3 % of AR ). All minor non-identified degradates accounted for m aximum 2.1 % of AR.

Table B.8.1.1.1-8: Distribution and recovery of app lied radioactivity [% of AR] after application of [3- 14 

### C] labelled spirotetramat to two soils under realis tic outdoor conditions.

Soil DAT Extractable CO 2 /volatiles NER Total

1 69.2 na 4.6 73.8

7 63.9 na 9.8 73.7

14 54.6 na 7.9 62.5
28 46.8 na 13.2 60.0
63 34.9 na 8.4 43.3 ‘Laacherhof AIIIa’

127 16.4 na 8.5 24.9

1 87.2 na 7.3 94.6

7 64.5 na 15.2 79.7

14 62.2 na 10.6 72.8
28 50.6 na 14.6 65.2
63 29.1 na 16.8 46.0 ‘Molino’

127 17.4 na 5.4 22.8 na denotes not applicable for this kind of study (‘ open system’)– 22 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.1.1-9: Characterization of extractableradioactivity [% of AR] after application of [3-14C] labelled spirotetramat to two soils under realisticoutdoor conditions.Dose DAT SpirotetramatSpirotetramat-enolSpirotetramat-ketohydroxySpirotetramat-MA-amideSpirotetramat-glyoxylicamideDimethyl-benzoic acidSpirotetramat-enol-dimer 1Spirotetramat-enol-dimer 2Unidentifieda153.6 2.5 6.6 nd 0.8 1.0 1.1 nd 3.6713.45.922.7 2.62.3 3.31.30.911.6143.8 3.323.63.6 1.6 3.0 1.4 0.6 13.7282.5 1.510.56.21.1 2.11.50.7 20.8632.5 1.014.2 2.0 nd 0.4 0.4 0.3 14.1‘LaacherhofAIIIa’1271.0 0.5 1.9 2.8 nd 0.5 nd nd 9.7172.2 3.85.0 0.2 0.2 0.3 0.8 0.3 4.2717.87.820.4 1.8 0.6 1.2 1.10.913.01410.25.425.31.90.7 2.10.8 0.4 15.4284.5 1.920.5 2.0 0.6 0.91.50.6 18.3631.4 0.9 3.14.9 0.71.0 0.2 0.2 16.7‘Molino’1271.0 0.48.0 1.0 < LOQ 0.3 < LOQ < LOQ 6.6nd denotes not detectedLOQ denotes limit of quantificationaUp to 29 detected regions, values for a single non-identified region were < 2.1% of AR

### – 23 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.1.1-10: Calculated DT 50 

### and DT 90 

### values [days] of [3- 14 

### C] labelled spirotetramat in two soils under realistic outdoor conditions based on S FO and DFOP (best fit) kinetics.
Application rate [g ha -1 ] Soil DT 50 [days] DT 90

[days] R 2 Chi 2 error [%] Kinetics a

1.2 4.1 0.981 15.4 SFO ‘Laacherhof AIIIa’ 1.1 8.8 nc 3.7 DFOP b

2.9 9.6 0.990 9.9 SFO 288
‘Molino’ 2.2 13.6 nc 1.5 DFOP b

a For modelling purposes 0-DAT values were set to 10 0.0
b DFOP kinetics data not stated in the study report, calculated by the reviewer



### Conclusion: The outdoor metabolism study in two semi-disturbed soils (which were already used within the laborator y degradation study) was conducted as supportive stud y. The purpose of this study was to test the degradation pattern of spirotetramat under more nat ural conditions. Further, the formulated radiolabel led spirotetramat was used (volume sprayed onto the soi l surface). It was shown that spirotetramat was qui ckly degraded. Following simple first order kinetics (SF O), calculated DT 50 

### values were 1.2 and 2.9 days for ‘Laacherhof AIIIa’ and ‘Molino’ soil, respectively. Following best-fit kinetics (DFOP), DT 50 

### values were 1.1 and 2.2 days, respectively, with DT 90 

### values of 8.8 and 13.6 days. A conservative estima te of first-order halflives for spirotetramat calculated from the best-fi t kinetics (i.e. DFOP-DT 90 

### divided by 3.32) yielded 2.7 and 4.1 days, respectively. Release of CO 2 

### was not measurable since the study was designed as ‘open outdoor’ study. Mineralization could only be addressed by ca lculation of the losses of radioactivity (including possible translocation of radioactivity into deeper soil lay ers whereas no drainage water was obtained during t he study). Within the study > 75 % of AR was lost unti l study termination (DAT-127). The NER fraction increased up to 13.2 % and 16.8 % of AR in the two soils by DAT-28 and by DAT-63, respectively. Beside a large number of minor non-identified degradates (no ne exceeding 2.1 % of AR individually), one major metabolite was detected: spirotetramat-ketohydroxy (max. 25.3 % of AR by DAT-14). Spirotetramat-enol w as detected with amounts > 5 % of AR at two successive DATs in ‘Molino’ soil (max. 7.8 % of AR by DAT-7). Degradates as spirotetramat-MA-amide, dimethyl-benz oic acid, spirotetramat-glyoxylic-amide, spirotetra matenol-dimer 1, spirotetramat-enol-dimer 2 and spirot etramat-ketohydroxy-carboxy were identified as mino r degradates. In comparison to the laboratory degrada tion study three new metabolites were identified wi thin this study: spirotetramat-glyoxylic amide, dimethyl -benzoic acid and spirotetramat-ketohydroxy-carboxy

, whereas the first two also were identified within t he laboratory photolysis study and now confirmed to occur under outdoor conditions as well. Further it has to be concluded that the formation of the spirotetram at-enoldimers within the outdoor study with another applic ation technique (broadcast application) was about < 1.5 %
of AR.

Reviewer comments:

• Study will be used as additional information, only

.

• The considerable lower formation of NER is in stro ng contrast to findings in the parent lab study and the spirotetramat-enol study. For this reason a sta tement was requested by the reviewer.

Notifier’s (BCS) response (position paper Hellpoint ner et al., 2007): Several reasons are expected to lead to reduced NER formation in the 14 

### C-parent outdoor study as compared to the parent lab aerobic soil study:

In the lab studies sieved soils (2 mm particle size) were used in contrast to semi disturbed (nonsieved, stamped) soils in the outdoor study. As a c onsequence, lab soil samples consist of particles with exaggerated surface area on the top and in the upper soil layer compared to the outdoor soils with reduced surface area particles. This will lead to an increased likelihood of applied test item to react with soil surface compon ents (chemisorption) in the lab study. Slightly mechanical soil mixing after application in the lab study is expected to increase this tendency in

### – 24 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

contrast to the outdoor soils treated with formulat ed test item, but untreated mechanically
afterwards.

More surface-located test item in combination with improved outdoor biomass conditions and increased outdoor aerobicity would be expected to r esult in increased enol degradation (oxidation), ketohydroxy formation and finally high er mineralization. The latter is expected to be the reason for the increasing radioactivity balance gaps of this non-balance outdoor study towards the study end.

The described processes are not expected at all to lead to increasing metabolite leaching potential but rather to an increased mineralization from the top soil. This is in line with the results from the US terrestrial field dissipation s tudies were no leaching potential of the soil analytes was observed (parent, enol, ketohydroxy, M A-amide).

### – 25 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: [Azaspirodecenyl-3- 14 

### C]- and [Azaspirodecenyl-5- 14 

### C]-Labelled BYI08330- cis

Enol: Comparative Aerobic Soil Metabolism/Degradati on in Four Soils Author(s), year: Babczinski, P., 2006b Study/report number: M-269304-01-2, MEF-05/157 Guideline(s): EC-Directive 91/414/EEC Annex I Part 7 and Annex II Part 9, OECD Guideline 307, EPA-Guidelines for Aerobic Soil Metabolism Studies, § 162-1, Japanese MAFF Guidelines
GLP: Yes
Deviations: None Validity: Study considered acceptable

Purpose of study: In the aerobic metabolism/degradation study with sp irotetramat it was not possible to determine the ki netics of all degradates since the main degradate spirotet ramat-enol showed a partial instability under the a cidic extraction/processing conditions required for spiro tetramat stability. Further it was assumed that irr egular spirotetramat-enol levels due to artifactual high s pirotetramat-enol-dimer formation (caused by “hot s pot” application technique) did not allow for adequate d egradate kinetic evaluations. Therefore, this separ ate spirotetramat-enol metabolism/degradation study was performed.

Material and methods:
Test substance: • [Azaspirodecenyl-3- 14 

### C]-spirotetramat-enol : sample ID: BECH 00992; specific activity 4.54 MBq mg -1 

### (122.8 μCi mg -1 

### ); radiochemical purity: > 99 % (acc. radio-HPLC and -TLC), identity and pur ity of the stock
solutions were checked

• [Azaspiro-decenyl-5- 14 

### C]-spirotetramat-enol : sample ID: BECH 00995; specific activity 4.99 MBq mg -1 

### (134.94 μCi mg -1 

### ); radiochemical purity: > 99 % (acc. radio-HPLC and -TLC), identity and pur ity of the stock
solutions were checked Reference substances: Spirotetramat-enol (unlabelle d), spirotetramat-ketohydroxy, spirotetramat-MAamide, spirotetramat-enol-dimer 1 Test system: Static incubation test system Test conditions: Study was performed in static incu bation test systems under aerobic conditions in the dark at 20 ± 1 °C. Moisture adjustment after ap plication to either 60 % MWHC (EU soils) or approx. 80 % of 1/3 bar moisture (US soil). At DAT-4 the absolute microbial biomass carbon contents corresponded to 4 .7, 6.0, 2.7 and 1.3 % relative to the soil organic carbon content of the soils ‘Hoefchen’, ‘Laacherhof AXXa’, ‘Laacherhof AIII’ and ‘Molino’, respectively

. For all four soils and all sampling intervals, the two test flasks containing the soils treated with the two differently labelled test item s were considered as replicates. In addition, for DAT-0 samples separate replicates were set up with the [5- 14 

### C]
labelled test item. Test duration: 119 days; sampling intervals: 0, 0.2 5, 1 ,4, 7, 14, 32, 60, 90 and 119 days Application: A higher volume (7.4 to 11 mL) of aque ous application solution for the treatments were used considering that the application techniqu e might have an influence on the metabolic pattern (see Babczinski, P., 2005) US soil – actual rate of [3- 14 

### C]-spirotetramat-enol applied was 53.5 kBq flask -1



### , corresponding to 118 μg kg -1 

### soil (DM); actual rate of [5- 14 

### C]-spirotetramat-enol applied was 62.9 kBq flask -1 

### corresponding to 126 μg kg -1 

### soil (DM), respectively.

### – 26 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

EU soils – actual rate of [3- 14 

### C]-spirotetramat-enol applied was 133.8 kBq flask -1



### , corresponding to 295 μg kg -1 

### soil (DM); actual rate of [5- 14 

### C]-spirotetramat-enol applied was 139.9 kBq flask -1 

### , corresponding to 280 μg kg -1 

### soil (DM), respectively. The calculations were based on homogeneous distribu tion in 15 cm depth of US soil, and in 2.5 cm depth of EU soils. Analyses: Aqueous extraction method : 100 g dry (DM) soil samples of each sampling
interval were extracted once with 80 mL of water co ntaining 0.025 % NH 3 

### at room temperature (aqueous ammonia, resulting in pH value s of approx. 7.0 to 8.5). Ambient extraction procedure : The residual soil after aqueous extraction was
extracted three times with 80 mL of acetonitrile/wa ter (1:1, containing 0.1 % ammonium chloride [w/v] and 0.06 % ammonia [v/v]; p H approx. 8.5) by shaking at room temperature. Aggressive extraction procedure : This was followed by an “aggressive” microwave extraction at 70 °C (10 min) using the same medium as for the conventional
extractions. The spirotetramat-enol residues were radio-assayed by LSC and analyzed separately in the aqueous extract, the combined amb ient extracts and the “aggressive” extract by reversed phase (C-18) HPLC. Solid samples (i.e. soil and paper filters) were combusted and 14 

### C levels were measured using LSC. For identification of test item and transformation prod ucts, co-chromatography and spectrometric methods were used. Volatile traps: Trap attachment (permeable for oxyg en) containing soda lime for absorption of

14 

### CO 2 

### and a polyurethane foam plug for adsorption of vol atile organic compounds (identification of 14 

### C as 14 

### CO 2 

### by BaCO 3 

### precipation) Analytical techniques: LSC, RP-HPLC, Si-60 silica T LC, LC-MS
Detection limits: LSC: LOQ ≤ 0.2 % of AR, for CO 2 

### samples LOQ < 0.1 % of AR; HPLC: LOQ ≤ 0.3 % of AR. Kinetic evaluation: Best fit kinetic model: SFO, FO MC and DFOP; MatLAB, vers. 7.0.4.365; LOD was set to 0.1 % of AR; values below the LOD were repor ted as < 0.1 % of AR.

### – 27 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.1.1-11: Main soil properties.

Parameter ‘Molino’ ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’

Source Florida, USA Monheim,

Germany Monheim,

Germany Burscheid,

Germany Texture (USDA) Sandy loam Sandy loam Silt loam Silt

Soil taxonomic classification (USDA) Fine-loamy, kaolinithic, thermic Plinthic Kandiudults Sandy, mixed,

mesic Typic Cambudolls Loamy, mixed,

mesic Typic Argudalfs Loamy, mixed,

mesic Typic Argudalfs

SCS soil series Ultisol Mollisols, Udolls Alfisols, Udalfs Alfisols, Udalfs

Sand (USDA) [%] 77.3 72.4 36.9 8.5 Silt (USDA) [%] 12.7 22.6 51.1 81.3 Clay (USDA) [%] 10.0 5.0 12.0 10.2 pH (H 2 O) 6.1 7.2 7.4 7.6 pH (CaCl 2 ) 5.4 6.3 6.5 6.7 pH (KCl) nd 6.4 nd 7.0 Organic matter [%] 1.60 1.75 1.43 3.63 Organic carbon a [%] 0.93 1.02 0.83 2.11 Carbonate (CaCO 3 ) [%] < 0.1 < 0.1 < 0.1 0.9 CEC [meq/100 g DM] 6 8 8 15 WHC at pF2.5 (FC at 0.33 bar) [g/100 g DM] 18.4 29. 1 20.7 35.6 MWHC [g/100 g DM] Not available 34.4 36.4 63.1
Microbial C b [mg microbial C/kg DM] 21 days before application 103 627 363 1012

Microbial C b [mg microbial C/kg DM] DAT-4 118 613 226 1001

Microbial C b [mg microbial C/kg DM] DAT-62 native: 88 with solvent: 78 native: 478

with solvent: 422 native: 159

with solvent: 184 native: 686

with solvent: 712

Microbial C b [mg microbial C/kg DM] DAT-119 native: 55 with solvent: 55 native: 366

with solvent: 374 native: 123

with solvent: 77 native: 572

with solvent: 565

nd denotes not determined

a Organic C = organic matter / 1.724
b Substrate induced respiration



### Findings: The DAT-0 recovery showed that for the ‘Molino’ and ‘Laacherhof AIII’ soils the extraction efficacy ra nged between 92.0 % and 103.2 %, 4.2 % to 9.3 % of AR we re still bound on the soil matrix. For the ‘Laacher hof
AXXa’ and ‘Hoefchen’ soils 23.4 % to 28.4 % of AR w as recovered as bound residues and only 76.0 % to
77.6 % of AR were extractable. It was shown that th e degradation of spirotetramat-enol was found to be ongoing during sample processing, starting with the aqueous extraction. Already immediately after application (1 – 2 minutes) spirotetramat-enol oxid ation was leading to spirotetramat-ketohydroxy.

The material balance ranged from 84.8 % to 108.4 % of AR (both labels), with three out of 80 values ra nged below 90 % at 84.8 %, 88.9 % and 89.2 %; one flask was at 76.4 % most probably due to volatile leakage and was rejected.

At study termination the amount of 14 

### CO 2 

### accounted for 16.7 % to 27.8 % of AR when using th e [5- 14 

### C] labelled test item. Higher mineralization levels we re expected with the [3- 14 

### C] test item accounting for 28.2 % to 43.0 % of AR until DAT-119. No volatile organics were evolved from the systems.

The non-extractable residues (NER) ranged from 4.2 % to 28.4 % of AR ([5- 14 

### C] label) and from 8.7 % to 23.8 % of AR ([3- 14 

### C] label) at DAT-0, respectively and increased imme diately up to 47.6 % - 59.6 % of AR

### – 28 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

except of ‘Laacherhof AIII’ soil (NER 27.9 / 26.1 % of AR) at DAT-0.25. At the termination of the stud ies
bound residues reached a level of 44.8 % to 62.5 % of AR (both labels). The non-extractable 14 

### C-residues were lower in the [3- 14 

### C] label experiments as compared to the [5- 14 

### C] label. The same trend was shown by the extractable residues. A partitioning of the het erogeneous NER radioactivity into humic acid, fulvi c acid and humin like fractions was observed, with general ly a major portion of radioactivity in the fulvic a cid fraction at later sampling intervals and with a significant portion still remaining bound to the soil in the in soluble humin
fraction.

Using the aqueous extraction step prior to the conv entional organic/aqueous extraction a significant p art of AR could only be dissolved at the very early DAT (d espite a solubility of 78 mg spirotetramat-enol per litre of water). Extractable 14 

### C residues were > 90 % of AR at DAT-0 for soils ‘Mo lino’ and ‘Laacherhof AIII’ and
between 76.0 % and 77.6 % of AR for soils ‘Laacherh of AXXa’ and ‘Hoefchen’. A decrease to < 25 % of AR in all four soils was observed, ranging from 9.2 % of AR (soil ‘Hoefchen’, [5- 14 

### C] label) to 24.7 % of AR (soil ‘Molino’, [3- 14 

### C] label) at study termination.

During the study a great number of radioactive peak s were detected and quantified. Under unidentified radioactivity all minor peaks plus diffuse radioact ivity was summed up. In the Table B.8.1.1.1-14 all components are given as mean values (except of the unidentified) whereas the two labels were considere d as duplicates. This was only applicable where the r espective component still carried both labels withi n the
molecule.

The test item spirotetramat-enol showed a biphasic decline whereas in the first phase a very fast degr adation was observed (in three soils a degradation of > 80 % of AR within six hours). This phase was matched b y a rapid appearance of the major degradation product s pirotetramat-ketohydroxy. The main metabolite spirotetramat-ketohydroxy accounted for max. 24 % of AR at DAT-1 in soil ‘Laacherhof AIII’. The spirotetramat-enol-dimer 1 (max. 5 % of AR at DAT-60) amounted once to 5 % of AR in one soil and spirotetramat-enol-dimer 2 did not reach the 5 % during the study. Spirotetramat-MA-amide amounted once > 5 % of AR ( 5.2 % of AR at DAT-4). Two further minor metabolites and spirotetramat-desmethylenol (max. 2 % of AR at DAT-4) were identified. In addi tion, as a previously unknown metabolite, spirotetramat-oxo-ketohydroxy (<< 0.5 % of AR), was identified.

A correlation between the biomass content and the d ecline of spirotetramat-enol and the formation of spirotetramat-ketohydroxy and spirotetramat-MA-amid e seems to exist. Further, no correlation was found between decline of spirotetramat-enol and the pH va lue of the soil.

Owing to the visual inspection and the statistical results (chi 2 

### error, R 2 

### ) the SFO model was not the appropriate kinetic model for this substance. In a second step the bi-phasic model FOMC (First-order m ulti compartment) according to Gustafson-Holden showed m uch better results. But the most appropriate model was the DFOP model (double first order in parallel) . The calculated DT 50 

### values using the DFOP model ranged from 0.02 – 0.2 days. The respective DT 90 

### values were in a range from 22.9 – 106.9 days. A recalculation of the degradation kinetic of spirote tramat-enol was conducted (see point B.8.1.2.3, Röp ke, 2006a).

The degradation behaviour of the metabolites spirot etramat-ketohydroxy and spirotetramat-MA-amide was recalculated by a separate study (B.8.1.2.3, Röpke, 2006a).

The minor metabolite spirotetramat-desmethyl-enol a nd the minor “metabolites” spirotetramat-enol-dimer 1 and 2 were not considered kinetically.

### – 29 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Metabolic pathway The degradation pathway of spirotetramat-enol confi rms the pathway already explained in the spirotetra mat metabolism study. In the major pathway the spirotet ramat-enol will be oxidized at the benzylic carbon position into spirotetramat-ketohydroxy which is hy drolytically opened into the spirotetramat-MA-amide and finally mineralized into CO 2 

### . In a second (minor) pathway, demethylation of the spirotetramat-enol into spirotetramat-desmethyl-enol is followed by entire mineralization to CO 2 

### . In a third (minor) pathway spirotetramat-ketohydroxy is demethylated leading t o the spirotetramat-desmethyl-ketohydroxy and mergi ng into spirotetramat-oxo-ketohydroxy by oxidation fol lowed by mineralization to CO 2 

### . In a fourth degradation pathway the spirotetramat-enol-dimer 1 and spirotet ramat-enol-dimer 2 were pretended to be built throu gh oxidative enol dimerization, but this pathway is re garded as non relevant under environmental conditio ns (artifactual character). These results were include d in the proposed overall metabolic pathway of spirotetramat in soil (see Figure B.8.1.1.4-1).

### – 30 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.1.1-12: Distribution and recovery of a pplied radioactivity [% of AR] after application of [3- 14 

### C] and [5- 14 

### C] labelled spirotetramat-enol to four soils incuba ted under aerobic conditions.

Total Extracted NER CO 2 Total Soil DAT [5- 14 C] [3- 14 C] [5- 14 C] [3- 14 C] [5- 14 C] [3- 14 C] [5- 14 C] [3- 14 C]

0 92.0 99.7 9.3 8.7 na na 101.3 108.4
0.25 50.3 48.6 48.5 50.5 0.5 1.5 99.4 100.6

1 46.1 49.8 50.7 50.3 1.0 3.3 97.8 103.5

4 44.0 44.2 50.2 50.3 1.9 6.1 96.1 100.6

7 45.7 43.7 48.4 46.9 2.8 7.9 96.9 98.5

14 39.9 41.0 52.6 49.6 4.2 11.2 96.6 101.8
32 36.8 34.3 52.4 49.4 7.6 17.1 96.8 100.8
60 31.5 28.2 52.7 49.8 10.9 20.1 95.1 98.1
90 37.2 33.2 45.2 42.7 14.0 24.8 96.3 100.7 ‘Molino’

119 24.7 20.3 55.3 52.3 16.7 28.2 96.8 100.8

0 76.0 77.6 28.4 23.6 na na 104.4 101.2
0.25 48.8 50.2 49.7 47.6 0.4 1.3 98.8 99.1

1 48.9 49.4 50.2 48.5 1.0 3.5 100.1 101.3

4 46.0 43.8 46.2 51.6 2.4 8.2 94.6 103.6

7 44.9 41.1 51.1 44.0 3.7 12.3 99.8 97.5

14 38.9 34.5 52.9 51.1 5.5 17.4 97.3 103.1
32 33.9 27.9 54.7 47.6 10.2 23.4 98.8 98.9
60 30.9 24.9 44.9 38.9 13.4 26.5 89.2 90.3
90 35.9 27.3 36.3 40.8 16.8 31.1 88.9 99.2 ‘Laacherhof AXXa’

119 22.9 18.5 54.3 45.5 19.0 32.5 96.2 96.5

0 103.2 99.6 4.2 4.4 na na 107.4 103.9
0.25 72.0 73.3 27.9 26.1 0.2 0.3 100.2 99.7

1 57.6 55.8 43.4 40.9 0.8 2.3 101.9 99.1

4 48.8 48.4 49.6 44.2 2.1 5.8 100.6 98.4

7 47.3 43.2 50.4 45.7 2.9 9.1 100.6 98.0

14 38.9 36.2 56.3 45.0 6.0 16.0 101.2 97.2
32 29.3 24.5 57.8 46.5 11.6 25.8 98.7 96.8
60 21.4 16.9 58.3 46.6 19.2 34.9 98.9 98.4
90 21.4 14.6 53.3 43.1 23.4 40.3 98.1 98.0 ‘Laacherhof AIII’

119 12.8 9.9 57.0 44.8 27.8 43.0 97.5 97.7

0 77.1 76.0 25.6 23.8 na na 102.7 99.8
0.25 38.8 40.3 59.6 54.5 0.3 1.5 98.8 96.3

1 40.0 39.1 57.9 53.3 1.3 5.3 99.2 97.7

4 33.8 30.1 60.1 52.7 3.8 12.4 97.7 95.1

7 29.3 24.8 60.9 53.7 5.7 17.1 95.9 95.5

14 24.6 19.7 59.3 56.7 8.6 8.6 92.6 76.4 a

32 20.9 17.1 63.0 52.5 12.6 27.2 96.5 96.7
60 17.1 12.7 62.0 51.1 17.2 31.9 96.3 95.8
90 19.8 13.9 44.8 41.3 20.3 35.2 84.8 90.4 ‘Hoefchen’

119 11.8 9.2 62.5 51.0 22.0 36.9 96.3 97.1

a Recovery loss caused by flask leakage– 31 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.1.1-13: Characterization [% of AR] of non-extractable soil residues (NER).Sampling DAT-0 DAT-14 DAT-32aDAT-32aDAT-60 DAT-60 DAT-119 DAT-119Soil ‘Hoefchen’‘LaacherhofAXXa’‘LaacherhofAXXa’‘Hoefchen’‘LaacherhofAIII’‘LaacherhofAIII’‘Molino’ ‘Molino’Total NER 49.7 48.0 53.6 62.5 57.7 44.2 54.3 44.1Fraction humic acids 19.1 18.7 20.9 19.1 9.2 6.2 9.6 9.0Fraction fulvic acids 19.2 17.4 20.6 28.8 28.0 23.835.8 28.0Fraction humin substances 15.4 10.0 11.7 18.7 19.214.6 10.2 8.3Processing balance [%] 108.0 95.9 99.2 106.6 97.7 101.0 102.3 102.8a‘DAT-30’ stated in the original report probably atypoTable B.8.1.1.1-14: Characterization of extractableradioactivity [% of AR] after application of [3-14C] and [5-14C] labelled spirotetramat-enol to foursoils incubated under aerobic conditions (means ofboth labels).UnidentifiedaSoil DATSpirotetramat-enolSpirotetramat-Desmethyl-enolSpirotetramat-enol-dimer 1Spirotetramat-enol-dimer 2Spirotetramat-ketohydroxySpirotetramat-MA-Amide[5-14C] [3-14C]078.0 nd 0.2 nd11.7 nd 5.4 6.50.2517.3 0.2 2.5 1.617.0 0.9 9.4 10.6113.4 0.5 1.8 1.717.41.4 10.9 12.5410.9 1.0 2.9 1.815.7 2.4 12.5 6.4714.71.11.2 1.412.0 2.9 14.7 7.91410.7 0.8 3.42.28.22.914.4 10.3327.7 0.93.61.65.4 2.2 16.1 12.2606.1 0.7 2.3 1.3 3.7 1.5 16.6 12.0907.1 0.7 2.3 1.3 3.8 1.321.9 15.5‘Molino’1194.8 0.4 1.8 1.8 2.5 0.9 11.4 9.4053.7 0.2 0.9 nd13.8 nd 7.2 9.40.2514.1 0.1 4.0 2.716.61.3 10.4 10.8114.7 0.5 2.4 3.016.2 2.6 10.9 8.8414.6 1.2 3.5 2.49.32.712.9 9.5717.01.61.8 2.56.0 2.0 13.8 10.51411.9 0.8 4.43.63.0 1.4 14.4 8.5328.7 0.4 4.9 2.8 2.2 0.1 14.4 9.0606.7 0.25.02.3 1.4 0.1 14.2 10.4907.6 0.3 4.1 2.0 1.8 0.220.4 10.8‘LaacherhofAXXa’1196.1 < LOQ 3.7 3.3 0.5 0.1 7.4 6.5‘Laacherhof 083.3 nd 0.3 nd11.2 nd 7.9 5.0– 32 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008UnidentifiedaSoil DATSpirotetramat-enolSpirotetramat-Desmethyl-enolSpirotetramat-enol-dimer 1Spirotetramat-enol-dimer 2Spirotetramat-ketohydroxySpirotetramat-MA-Amide[5-14C] [3-14C]0.2538.0 nd 1.4 1.022.3 0.9 10.3 7.9117.2 0.6 0.9 0.924.01.6 11.0 12.044.8b2.01.2 1.019.85.216.0 13.4711.6 1.6 0.7 1.211.1 4.9 13.814.6147.7 1.5 1.5 1.35.7 3.618.713.7325.7 0.4 1.81.32.2 0.9 16.0 13.5603.7 0.22.00.8 1.1 0.1 13.1 9.5904.0 0.1 1.0 0.6 0.9 0.2 14.6 7.9AIII’1192.7 nd 1.0 1.2 0.6 0.2 6.3 4.9051.6 nd nd nd18.4nd 5.9 7.10.2510.7 0.6 1.6 1.215.7 1.5 8.9 7.5110.5 1.3 1.1 1.413.32.58.210.7412.01.31.81.74.4 1.8 10.7 7.3711.8 1.0 0.9 1.5 1.9 0.7 10.4 7.9148.6 0.2 1.8 1.6 1.3 0.3 9.5 7.1326.3 0.2 1.9 0.9 1.4 0.2 9.8 6.2604.6 < 0.12.10.9 0.5 nd 9.2 4.3906.0 0.1 1.0 0.6 0.6 0.110.86.3‘Hoefchen’1193.8 nd 1.1 1.4 0.3 nd 4.6 3.3nd denotes not determinedaTotal % of AR for unidentified discrete componentsand streaking, none exceeding > 4.9 % of AR individuallybSpirotetramat-enol Dat-4 value was considered as outlier– 33 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.1.1-15: Characterization of extractableradioactivity [% of AR] after application of [3-14C] and [5-14C] labelled spirotetramat-enol to foursoils incubated under aerobic conditions (individual values for each label).SpirotetramatenolSpirotetramat-Desmethyl-enolSpirotetramat-enol-dimer 1Spirotetramat-enol-dimer 2Spirotetramat-ketohydroxySpirotetramat-MA-AmideUnidentifiedaSoil DAT[5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C]076.2 79.7 nd nd nd 0.5 nd nd10.313.0 nd nd 5.4 6.50.2518.7 15.8 nd 0.4 3.2 1.7 2.2 0.916.117.9 0.6 1.2 9.4 10.6113.3 13.5 0.4 0.6 1.8 1.8 1.9 1.516.918.01.0 1.9 10.9 12.549.2 12.5 0.7 1.33.62.3 2.2 1.414.017.3 1.8 3.0 12.5 6.4714.0 15.5 0.61.61.4 1.0 1.5 1.411.112.9 2.33.514.7 7.91410.3 11.1 0.3 1.2 3.2 3.6 2.22.16.99.42.53.3 14.4 10.3327.3 8.2 0.7 1.0 3.53.71.7 1.65.35.5 2.2 2.2 16.1 12.2605.8 6.31.00.5 2.1 2.5 1.5 1.1 3.1 4.3 1.4 1.6 16.6 12.0906.7 7.5 0.8 0.5 2.2 2.4 1.3 1.4 3.3 4.3 1.0 1.621.9 15.5‘Molino’1194.7 4.9 0.3 0.5 2.1 1.42.51.1 2.6 2.4 1.2 0.6 11.4 9.4055.0 52.3 nd 0.3 0.7 1 nd nd13.114.6 nd nd 7.2 9.40.2514.4 13.8 nd 0.3 4.3 3.8 3.4 2.115.417.80.9 1.7 10.410.8113.3 16.1 0.5 0.5 2.6 2.2 3.2 2.816.415.9 2.13.110.9 8.8415.0 14.3 0.9 1.5 3.7 3.3 2.8 2.18.510.12.33.1 12.9 9.5717.9 16.11.3 1.91.9 1.7 2.7 2.35.66.4 1.7 2.2 13.8 10.51411.7 12.0 0.8 0.8 4.44.53.63.72.7 3.4 1.2 1.7 14.4 8.5328.4 9.0 0.3 0.65.5 4.4 3.2 2.5 2.0 2.4 0.1 0.2 14.4 9.0606.5 6.9 0.3 0.15.74.2 2.8 1.7 1.2 1.6 0.1 nd 14.2 10.4906.9 8.2 nd 0.5 4.0 4.3 2.2 1.8 2.2 1.4 0.2 0.220.4 10.8‘LaacherhofAXXa’1196.6 5.6 <LOQ nd 4.4 3.13.92.7 0.7 0.4 nd 0.2 7.4 6.5086.1 80.6 nd nd 0.6 nd nd nd8.513.9 nd nd 7.9 5.00.2538.8 37.2 nd nd 1.6 1.1 1.2 0.819.425.20.6 1.1 10.3 7.9118.9 15.5 0.7 0.5 1.1 0.8 1.0 0.923.524.4 1.4 1.8 11.0 12.043.5b6.0b2.0 2.01.2 1.1 1.2 0.919.719.95.35.016.0 13.4712.9 10.2 1.2 1.9 0.8 0.61.50.912.010.35.2 4.7 13.814.6147.7 7.8 1.2 1.8 1.7 1.4 1.31.25.26.3 3.1 4.018.713.7326.1 5.3 0.3 0.52.21.4 1.6 0.9 2.3 2.1 0.8 0.9 16.0 13.5604.1 3.2 0.2 0.2 2.11.90.8 0.8 1.1 1.1 nd 0.3 13.1 9.5904.3 3.6 nd 0.1 1.1 0.9 0.5 0.6 0.8 0.9 nd 0.5 14.67.9‘LaacherhofAIII’1193.0 2.5 nd nd 1.3 0.8 1.4 1.0 0.5 0.6 0.2 0.2 6.3 4.9‘Hoefchen’ 053.8 49.3 nd nd nd nd nd nd17.319.6nd nd 5.9 7.1– 34 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008SpirotetramatenolSpirotetramat-Desmethyl-enolSpirotetramat-enol-dimer 1Spirotetramat-enol-dimer 2Spirotetramat-ketohydroxySpirotetramat-MA-AmideUnidentifiedaSoil DAT[5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C] [5-14C] [3-14C]0.2510.8 10.6 0.4 0.7 1.7 1.4 1.1 1.314.816.6 1.0 2.1 8.9 7.5113.7 7.3 1.31.31.2 1.1 1.5 1.311.914.82.3 2.78.210.7412.4 11.61.41.1 1.9 1.7 1.71.84.1 4.7 1.7 1.9 10.7 7.3712.9 10.8 0.9 1.0 0.9 0.8 1.5 1.6 2.0 1.9 0.7 0.8 10.4 7.9148.8 8.5 0.5 nd2.01.62.01.2 1.4 1.2 0.4 0.1 9.5 7.1326.5 6.1 0.2 0.32.01.8 0.8 1.1 1.4 1.4 0.1 0.2 9.8 6.2604.8 4.4 nd 0.12.0 2.30.8 1.0 0.4 0.6 nd <LOQ 9.2 4.3906.3 5.8 0.2 <LOQ 0.9 1.0 0.7 0.5 0.8 0.3 0.1 nd10.86.31194.1 3.5 nd nd 1.2 1.0 1.6 1.1 0.3 0.3 nd nd 4.6 3.3nd denotes not determinedaTotal % of AR for unidentified discrete componentsand streaking, none exceeding > 4.9 % of AR individuallybSpirotetramat-enol Dat-4 value was considered as outlier

### – 35 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.1.1-16: Calculated DT 50 

### and DT 90 

### values [days] of spirotetramat-enol in four aerobi c soils based on DFOP kinetics (best fit kinetics).

Soil DT 50 [days] DT 90 [days] R 2 Chi 2 error Kinetics

‘Molino’ 0.09 53.7 0.996 6.9 DFOP
‘Laacherhof AXXa’ 0.02 106.9 0.987 8.7 DFOP
‘Laacherhof AIII’ 0.20 22.9 0.996 7.2 DFOP
‘Hoefchen’ 0.02 74.0 0.993 8.1 DFOP



### Conclusion: Spirotetramat-enol ([3- 14 

### C] and [5- 14 

### C] labelled), the major degradate of spirotetramat, dissipates in soil following biphasic kinetics (best fit) and is thoro ughly mineralized. The calculated DT 50 

### values ranged from 0.02 – 0.20 days (DFOP kinetics), respective DT 90 

### values were in a range of 22.9 – 106.9 days. A pos itive correlation of the spirotetramat-enol dissipation a nd the formation of soil degradates with the soil b iomass was partly evident. No correlation was found betwee n spirotetramat-enol dissipation and the soil pH va lues. At study termination the evolved 14 

### CO 2 

### (no volatile organics) was about 16.7 % to 27.8 % of AR ([5- 14 

### C] labelled test item) and about 28.2 % to 43.0 % of A R ([3- 14 

### C] labelled test item), respectively. Up to > 50 % of
AR was detected in the NER fraction at DAT-1 and re ached a level of 44.8 % to 62.5 % of AR (both label s) at
DAT-119. Extractable residues decreased from > 90 % of AR at DAT-0 to a minimum of 9.2 % of AR at stud y termination.

Besides the test item spirotetramat-enol, spirotetr amat-ketohydroxy, spirotetramat-enol-dimer 1, spirotetramat-enol-dimer 2, spirotetramat-MA-amide, spirotetramat-desmethyl-enol and spirotetramat-oxo

ketohydroxy were identified. In all four soils, spi rotetramat-ketohydroxy was identified as major meta bolite (> 10 % of AR) with amounts up to 24 % of AR (DAT-1 ). All other degradates, with the exception of spirotetramat-MA-amide, were ≤ 5 % of AR (minor). No label-specific degradates we re observed throughout the entire study.

Reviewer comments:

• Labels were handled as replicates. Additional calc ulations by the reviewer confirmed that the degradation of both labels is almost identical. The refore the usage of a mean value is considered acceptable.

• The degradation behaviour of spirotetramat-enol, s pirotetramat-ketohydroxy and spirotetramat-MAamide was investigated in a separate kinetic analys is study (see B.8.1.2.3, Röpke, 2006a).

### – 36 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.1.2 Anaerobic degradation in soil (OECD Annex IIA, 7.1.2)

The route of anaerobic degradation (metabolism) of spirotetramat in soil was investigated in one study (also used for anaerobic degradation rate calculation):

• Aerobic/ anaerobic degradation in one soil (20 °C, [3- 14 

### C] label)

Reference: BYI08330: Anaerobic Soil Metabolism Author(s), year: Menke, U., 2006a Study/report number: M-270739-01-2, MEF-05/515 Guideline(s): OECD Guideline for Testing of Chemica ls, Guideline 307, Aerobic and Anaerobic Transformation in Soil, Adopted Document; US EPA Su bdivision N, Section 162- 2; CAN PMRA DACO 8.2.3.4.4; Japanese MAFF New Test Guidelines for Supporting Registration of Chemical Pesticides, 12 Nohsan 8147
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substances: [Azaspirodecenyl-3- 14 

### C]-spirotetramat; sample ID: BECH 0755; Specific ac tivity 3.71 MBq mg -1 

### (100.2 μCi mg -1 

### ); radiochemical purity: > 99 % (acc. Radio-HPLC and –TLC) Reference substances: Spirotetramat (unlabelled), s pirotetramat-enol, spirotetramat-ketohydroxy, spirotetramat-MA-amide, spirotetramat-di-hydroxy, s pirotetramat-oxo-enol, spirotetramat-enol-dimer 1 Test system: Static incubation test system
Test conditions: The study was performed for approximately one half- life of test item under
aerobic conditions in the dark at 20 ± 1 °C. Thereb y, the test system consisted of Erlenmeyer flasks (300 mL) attached with a trap att achment (permeable for oxygen) containing soda lime for absorption of 14 

### CO 2 

### and a polyurethane foam plug for adsorption of volatile organic compounds. Moisture adjustment after application to 50% of maximum water holding capacit y was carried out for each individual flask by addition of de-ionized water. Following the aerobic incubation phase (just 4.8 ho urs), the samples were flooded with oxygen-depleted de-ionized water (3 cm layer above soil level), set under nitrogen atmosphere, and maintained in the da rk under anaerobic conditions for 180 days at 20 °C. At start of the a naerobic study phase, the trap systems were replaced by sealable two-valve glass s toppers connected with plastic gas sampling bags, closing the flasks to ma intain their nitrogen atmosphere. Test duration: After 4.8 hours aerobic incubation p hase 180 days under anaerobic conditions Sampling intervals: Duplicate test flasks were proc essed for analysis after 4.8 hours of aerobic incubation, and at 0, 0.6, 1, 4, 6, 14, 32, 60, 90, 120 and 180 days after flooding the test systems (anaerobic incubation). Application: [3- 14 

### C]-spirotetramat was applied at an actual rate of 8 0.43 μg 100 g -1 

### soil DM (intended nominal concentration of 76.8 μg per flas k). This rate was equivalent to 105 % of the intended maximum field application rat e of 288 g ha -1 

### (assuming a homogeneous distribution in 2.5 cm topsoil layer). Analyses: Aerobic incubation : soils and traps were removed and subjected for ex traction and analysis

### – 37 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Anaerobic incubation : Firstly, flasks were connected to the volatiles c ombustion oven. After opening the flasks the water layers wer e measured for pH, redox potential and dissolved oxygen content. 30 mL of th e water layer was removed
for CO 2 

### analysis and the rest were separated from the soil layers by decanting and centrifugation and subjected to analysis.

The soil (aerobic and anaerobic) was extracted twic e with 80 mL of acetonitrile/water/formic acid (50:50:0.5, v/v/v) f ollowed by an extraction with 1 x 80 mL acetonitrile. After the organic extraction pr ocedure an acid extraction with twice of 80 mL of acetonitrile/1 M HCl (1:1, v/v) f ollowed. Organic and acid extracts were kept separate for individual chromato graphy profiling. Volume and radioactivity were determined by LSC. Extracted soils and paper filters were
combusted and 14 

### C levels were measured using LSC. Volatile compounds (contained in the PU foam plugs) were extracted with 25 mL of ethyl acetate and radio-assayed by LSC.
CO 2 

### absorbed by soda lime was liberated with HCl and p urged into LS cocktails with nitrogen and subjected to LSC. Volatile traps: Aerobic phase : Organic volatiles: polyurethane (PU) foam, CO 2 

### : soda lime. Anaerobic phase : Test system was closed with gas sampling bags, wh ich allowed for analysis of the gaseous headspace forme d upon anaerobic
incubation. Analytical techniques: LSC, RP-HPLC (C18), TLC, LC- MS, LC-MS/MS, NMR Detection limits: LSC: liquid samples LOQ ≤ 0.73 Bq, for CO 2 

### samples LOQ ≤ 1.03 Bq; HPLC:
LOQ ≤ 0.19 % of AR (9.26 Bq), LOD = 0.06 % of AR (1/3 of LOQ) Kinetic evaluation: SFO, FOMC and DFOP kinetics; Ma tLab 7.0.4.365., C 0 

### calculated by the model; used criterion for best fit model kinetic (to deriv e endpoints): chi 2 

### scaled error and visual assessment of the goodness of fit

Table B.8.1.1.2-1: Main soil properties.

Parameter ‘Laacherhof AIII’

Source Monheim, Germany Texture (USDA) Sandy loam Soil taxonomic classification (USDA) Loamy, mixed, mesic Typic Argudalfs
SCS soil series na Sand (USDA) [%] 52.45 Silt (USDA) [%] 30.90 Clay (USDA) [%] 16.65 pH (H 2 O) 7.4 pH (CaCl 2 ) 6.8 pH (KCl) 6.9 Organic matter [%] 0.69 Organic carbon a [%] 0.4 CEC [meq/100 g DM] 4.8 WHC at 0.33 bar (1/3 bar WHC) [g/100 g DM] 15.5 MWHC [g/100 g DM] 36.4
Microbial Biomass See remarks b

na denotes not available

a Organic C = organic matter / 1.724

b Measurement of initial soil biomass (638 mg microb ial C kg -1 soil DM) indicated that the soil was aerobic viabl e at the beginning of the study (treated with test item). Anaerobic bacteria plate count tests indicated that the soil was anaer obic viable throughout the anaerobic incubation phase.

### – 38 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Findings: Verification of the extraction and chromatographic procedures indicated that the chosen extraction met hod was well suitable and the recovery values of the HP LC method were in an acceptable range (101.8, 99.9 and 98.9 %).

The material balance ranged from 95.6 to 102.8 % (m ean 99.3 % ± 1.7 %).

The forming of 14 

### CO 2 

### during the aerobic incubation was rather low accou nted for max. 0.1 % of AR. The mineralization rate during the anaerobic incubation was in the same range with a maximum of 0.3 % of A R at DAT-90 (single value of replicate). Volatile organi c compounds were neither observed in the aerobic no r in the anaerobic phase (< 0.1 % of AR).

During the short aerobic incubation phase the NER fraction increased from 1.7 % of AR at test start (DAT-(- 0.2)) to 11.7 % of AR at day of flooding. At DAT-0. 6 the amount of non-extractable residues increased to its maximum of 17.5 % of AR. At study termination the b ound residues accounted for 7.9 % of AR (mean values).

The total radioactivity extractable from soil decre ased from DAT-(-0.2) from 99.8 % of AR to 32.1 % of AR at study termination. In contrast the total radioactiv ity extractable from the water layer increased from 24.6 % of AR on the day of flooding to an amount of 58.7 % of AR at DAT-180 (mean values). The unidentified radioactivity was summed up with the diffuse radioa ctivity.

The amount of the test item spirotetramat declined in soil (and entire system) from 95.5 % of AR to < 2 % of AR within 7 days. Beside the test item several meta bolites were identified. Spirotetramat-enol was identified as major metabolite in both layers with a maximum a mount of 27.9 % of AR (DAT-1) in soil, 43.0 % of AR (DAT-180) in the water layer and 54.6 % of AR (DAT- 180) in the entire system, respectively. Beside spirotetramat-enol, spirotetramat-ketohydroxy was identified as a second major metabolite. The m aximum
amount in soil was measured at the same time as for spirotetramat-enol (max. 15 % of AR, DAT-1). In th e water layer the maximum was reached with 6.2 % of A R at DAT-60. In the entire system spirotetramatketohydroxy increased from the maximum of 19.3 % of AR (DAT-1) to 7.7 % of AR at the end of the study. In addition, the following five degradates were identi fied. Spirotetramat-MA-amide amounted in soil to levels of up to 4.4 % of AR (DAT-1), in the water layer up to 5.0 % of AR (DAT-180) and therefore, in the ent ire system up to 7.2 % of AR at the end of the study. T he spirotetramat-enol-dimer 1 reached similar high levels of up to 7.0 % of AR (DAT-6) in the entire s ystem. In soil the highest level of up to 6.7 % of AR was
reached at DAT-6. Spirotetramat-enol-dimer 2 reached its maximum in soil of up to 4.6 % of AR a t the end of the study. Metabolites spirotetramat-di-hydroxy (max. 3.2 % of AR, DAT-32 and 90 in entire system, max. 1.8 % of AR, DAT-32 in soil) and spirotetramat-oxo-enol (max. 1.4 % of AR, not listed in table) were detected only at minor levels (mean values were giv en in all cases). Furthermore a metabolite designat ed
R30 was detected reaching levels of 8.2 % of AR at DAT -0.6 in soil and 8.3 % of AR in the entire system, but
did not occur with levels > 5 % of AR at two succes sive DATs. Attempts to fully elucidate its structur e by LCMS, LC-MS/MS and NMR were not successful. From the results of the LC-MS/MS spectrometric analysis it was concluded by the notifier that the major compon ent was an unknown enol based compound possibly originating from polymeric structures (like the eno l-dimers). All values from the entire system are su mmarized
in Table B.8.1.1.2-3.

Largest individual unknown metabolites reached up t o 1.8 % of AR in the soil layer and 1.7 % of AR in the water layer (single values).

The detected metabolites were almost identical to t hose formed upon the aerobic transformation in the aerobic metabolism study, except of the minor metab olite spirotetramat-di-hydroxy. Spirotetramat-di-hy droxy

### – 39 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

was also formed in the aerobic aquatic metabolism s tudy. Therefore it can be excluded that this metabo lite will be specially formed under anaerobic conditions

.

The kinetic analysis was done for the entire system (water and soil layer) during the anaerobic incuba tion phase. Three kinetic models (SFO, FOMC and DFOP) we re used to evaluate the best fit model. The FOMC
model indicated the lowest chi 2 

### error value and the best visual fit (see Table B.8 .1.1.2-4). The degradation rates of spirotetramat in the total system during t he anaerobic phase were calculated to be 0.06 days (DT 50 

### ) and 1.33 days (DT 90 

### ) with a R 2 

### of 0.993.

Table B.8.1.1.2-2: Distribution and recovery of app lied radioactivity [% of AR] after application of [3- 14 

### C] labelled spirotetramat under aerobic conditions, then turning to anaerobic conditions (mean of duplicate samples).

Soil DAT Water layer Extracted (organic, acid extract) CO 2 Volatiles NER Total recovery

-0.2 a na 99.8 na na 1.7 101.5

0 b na 85.2 0.1 na 11.7 97.0
0.6 24.6 56.5 0.1 < 0.1 17.5 98.7

1 23.1 58.5 0.1 < 0.1 17.2 99.0

4 33.7 47.4 0.2 < 0.1 16.3 97.6
6 33.7 50.0 0.2 < 0.1 15.5 99.4

14 40.0 47.5 0.2 < 0.1 12.4 100.1
32 48.6 39.7 0.2 < 0.1 12.5 101.0
60 51.4 36.8 0.2 < 0.1 12.2 100.5
90 52.6 34.6 0.2 < 0.1 11.6 99.1
120 55.2 32.9 0.1 < 0.1 10.7 98.9 ‘Laacherhof AIII’

180 58.7 32.1 0.1 < 0.1 7.9 98.8 na denotes not applicable

a Aerobic incubation phase (0.2 days)
b Flooding of the test systems– 40 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.1.2-3: Characterization of extractableradioactivity [% of AR] after application of [3-14C] labelled spirotetramat to one sandy loam soilincubated under aerobicconditions, then turning to anaerobic conditions –entire system(individual replicates).SpirotetramatSpiro-tetramat-di-hydroxySpiro-tetramat-MA-amideSpiro-tetramat-enolSpiro-tetramat-ketohydroxyR30Spiro-tetramat-enol-dimer 1Spiro-tetramat-enol-dimer 2UnidentifiedaSoil DATA B A B A B A B A B A B A B A B A B-0.2b94.9 96.2 < LQ < LQ < LQ < LQ 1.5 2.1 0.5 0.5 < LQ< LQ < LQ < LQ < LQ < LQ 2.2 1.80c55.2 62.8 < LQ < LQ 1.1 0.4 9.1 7.811.3 6.9 0.4 0.5 3.6 3.5 0.9 1.0 2.8 3.00.68.8 10.1 0.1 0.2 3.6 3.627.425.915.417.68.3 8.47.3 5.2 3.9 1.7 7.0 8.018.0 5.5 0.3 0.8 4.3 5.432.232.818.420.35.3 4.2 4.0 4.2 1.0 0.9 7.4 8.442.6 3.9 1.5 0.8 3.0 2.936.539.518.016.4 1.7 2.3 6.27.31.2 1.4 7.6 9.561.9 1.0 0.7 1.2 2.1 2.143.247.315.415.8 0.8 0.67.86.2 2.4 2.3 8.9 7.7142.1 1.0 2.2 1.9 1.6 1.953.549.712.916.3 0.7 0.2 4.0 4.2 2.8 3.4 8.3 8.3321.6 1.43.03.3 4.1 3.740.440.415.716.5 0.4 0.5 3.6 4.2 3.6 4.415.1 14.7601.5 1.4 2.3 3.0 4.7 4.345.148.114.416.1 0.3 0.2 1.0 0.9 3.2 2.9 14.4 12.6901.0 0.9 2.63.95.0 5.548.751.512.812.3 0.5 0.5 1.0 1.1 3.7 3.1 11.0 9.41200.9 1.03.03.3 6.7 6.347.946.711.511.5 0.4 0.3 0.6 0.54.14.0 13.3 14.0‘LaacherhofAIII’180< LQ < LQ 2.6 2.87.3 7.156.053.37.6 7.7 0.8 0.7 0.2 0.3 3.75.612.5 13.5LQ denotes LOQaUp to 28 unidentified regions, individual amounts< 1.8 % of AR in the soil and < 1.7 % of AR in thewater layerbAerobic incubation phase (0.2 days)cFlooding of the test systems

### – 41 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.1.2-4: Calculated DT 50 

### and DT 90 

### values [days] of spirotetramat in ‘Laacherhof AIII ’ soil incubated under anaerobic conditions – entire syste m (SFO, FOMC and DFOP kinetics). Soil Compartment DT 50 [days] DT 90 [days] Chi 2 error Kinetics

0.25 0.83 17.95 SFO
0.06 1.33 5.27 FOMC ‘Laacherhof AIII’ Entire system 0.22 0.87 9.23 DFOP



### Conclusion: Under anaerobic conditions spirotetramat declines q uickly with DT 50 

### values of < 1 day in the total system. The two major metabolites spirotetramat-enol and sp irotetramat-ketohydroxy occurred with amounts > 10 % of AR in the entire system. Spirotetramat-enol was rather stable during the anaerobic incubation perio d, spirotetramat-ketohydroxy increased to 19.3 % of AR and decreased < 10 % of AR towards DAT-180. Spirotetramat-MA-amide reached levels > 5 % of AR t owards the end of the study and showed increasing tendency. Spirotetramat-enol-dimer 1 reached its ma ximum during the first 7 days afterwards decreasing to 0.3 % of AR at the end of the study. The second for med dimer, spirotetramat-enol-dimer 2 increased tow ards the end of the study and reached levels up to 4.6 % of AR. During the incubation period the mineraliza tion
was rather low.

The metabolites found in the soil and water layers were almost identical to those identified in the ae robic metabolism study and in the abiotic hydrolysis expe riments except of spirotetramat-di-hydroxy. Spirotetramat-di-hydroxy was formed in the aerobic aquatic metabolism study, therefore it can be concl uded that this metabolite will not specially occur under anaerobic conditions. The unknown metabolite R30, which was not identified by the notifier, is not a releva nt metabolite since the measured levels only once e xceeded the 5 % of AR trigger (EU risk assessment).

Comment (reviewer):

• CO 2 

### identification was not proven owing to the low amo unts formed in this study and the high overall recovery. This is accepted by the reviewer.

### – 42 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.1.3 Soil photolysis (OECD Annex IIA, 7.1.3)

Soil photolysis was investigated in one study:

• Soil photolysis on one soil, [3- 14 

### C] and [5- 14 

### C] labelled spirotetramat

Reference: [Azaspirodecenyl-3- 14 

### C]- and [Azaspirodecenyl-5- 14 

### C]-BYI08330:
Phototransformation on Soil Author(s), year: Heinemann, O., 2005a Study/report number: M-252907-01-2, MEF-04/481 Guideline(s): Commission Directive 95/36/EC amending Council Dire ctive 91/414/EEC, 1995 Pesticide Assessment Guidelines, Subdivision N, Env ironmental Fate, US EPA, 161-3: Photodegradation Studies on Soil, 1982 Canada PMRA, DACO No. 8.2.3.3.1 Considering Draft (2004-02) OECD Guideline for the Testing of Chemicals:
Photo-transformation of Chemicals on Soil Surfaces
GLP: Yes
Deviations: None Validity: Study considered partly valid, only (refe r to comments)

Material and methods:
Test substances: • [Azaspirodecenyl-3- 14 

### C]-spirotetramat: sample ID: BECH 1518; specific activity 3.71 MBq mg -1 

### (100.2 μCi mg -1 

### ); radiochemical purity: > 99 % (acc. Radio-HPLC and – TLC); identity and purity of the stock solutions was checked

• [Azaspiro-decenyl-5- 14 

### C]-spirotetramat: sample ID: BECH 1519; specific activity 4.03 MBq mg -1 

### (108.8 μCi mg -1 

### ); radiochemical purity: > 98 % (acc. Radio-HPLC and – TLC); identity and purity of the stock solutions was checked Reference substances: Spirotetramat (unlabelled), s pirotetramat-enol, spirotetramat-ketohydroxy, dimethyl-benzoic acid. Test system and experimental design: Spirotetramat was applied using both labels at a ra te of 1.9 mg ai kg -1 

### soil, (equivalent to 288 g ai ha -1 

### assuming 1 cm soil depth and 1.5 g cm -3 

### soil density) as drops across the soil surface. Duplicate samples were maintained at 20 ± 1 °C and irradiated for 7 days. Glass flasks filled with 3 g DM of viable soil (3 m m thickness), closed by a quartz glass cover and attached with a trap attachment. Mo isture adjustment after application to 75 % of 1/3 bar moisture was carried out and adjusted in both irradiated and dark samples at each sampling interv al. Control samples were incubated in the dark.
A supplemental

(non-kinetic) experiment (DAT-0 and DAT-7 sampling only)
was conducted with dried soil ‘Molino’ and a second moist soil (‘Laacherhof AIII’) to identify soil specific and/or moisture effects. For metabolite identification two test systems were treated at 10-times of the application rate. Irradiation: Continuously artificial irradiation us ing light from a xenon arc lamp (xenon lamp with < 290 nm cut-off filter, approx. 1115 W m - 

### 2 for [3- 14 

### C] and 1132 W m - 

### 2 for [5-

14 

### C] label). The experimental light intensity of 7 da ys continuous irradiation is equivalent to 37 solar days in June under extreme s unlight conditions at Phoenix, AZ (USA, 33 °N) or to 59 days in June under extreme European conditions in

### – 43 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Greece (Athens, EU, 38 °N).

Sampling intervals: 0, 0.2, 1, 2, 3, 4, and 7 days Supplemental experiment: soil ‘Laacherhof AIII’ sam plings were conducted at DAT-0 and DAT-7, only Soil handling: Air-dried, sieved to 2 mm, stored at 4 °C Analysis: Soil samples were extracted with 5 mL of acetonitrile/water (1/1, v/v, 0.5 % formic acid), two times with 5 mL of acetonitrile/1 M aqueous hydrochloric acid (1/1, v/v), and once with 5 mL of acetonitrile. Rad ioactivity in supernatants were analysed by LSC and analyzed separately in the comb ined organic and combined acidic extracts by RP-HPLC with radioactiv ity detection. Solid samples (i.e. soil and paper filters) were combusted and 14 

### C levels were measured using
LSC. Volatile traps: Organic volatiles: polyurethane foa m, CO 2 

### : soda lime. Analysis techniques: LSC,HPLC; HPLC-MS, HPLC-MS/MS, TLC, MS (ESI), NMR Detection limits: HPLC: LOQ = 0.96 % of AR, TLC: LO Q = 1.3 % of AR. Kinetic evaluation: Simple first order (SFO) kineti cs; ModelManager, vers. 1.1.; arithmetic means were used for the mentioned replicates

Table B.8.1.1.3-1: Main soil properties.

Properties ‘Molino’ ‘Laacherhof AIII’ (supplemental test) Texture (USDA) Tifton fine sandy loam Loam

Soil taxonomic classification (USDA) Fine-loamy, kaolinitic, thermic, plinthic, kandiudults Loamy, mixed, mesic typic Argudalfs

SCS soil series Ultisol Alfisols Sand (USDA) [%] 77.3 49.0 Silt (USDA) [%] 12.7 31.0 Clay (USDA) [%] 10.0 20.0 pH (H 2 O) 6.1 7.1 pH (CaCl 2 ) 5.4 6.5 Organic matter [%] 1.6 2.1 Organic carbon a [%] 0.93 1.2 CEC [meq/ 100 g DM] 6 10.2 Field moisture capacity at 0.33 bar [g water/100 g DM] 8.8 17.1 Microbial Biomass initial [mg microbial C/kg DM] 96 na

a Organic C = organic matter / 1.724 na not applicable

### – 44 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Findings: The average material balances for the irradiated test systems ranged from 95.2 % to 101.7 % of AR ([3-

14 

### C] label) and from 95.9 % to 104.7 % of ([5- 14 

### C] label). The extractable residues decreased from an average of 99.5 % and 103.0 % of AR at DAT-0 to 81. 7 % and 76.5 % of AR at DAT-7 for [3- 14 

### C] and [5- 14 

### C]
label. The rates of non-extractable residues increa sed from 0.1 % of AR (both labels) at day 0 to 9.6 % of AR ([3- 14 

### C] label) and 12.1 % of AR ([5- 14 

### C] label) at the end of the study. The 14 

### CO 2 

### formation was considerably higher in the irradiated system as in the dark cont rols and reached its maximum at DAT-7 with rates of about 3.8 % ([3- 14 

### C] label) and 7.3 % ([5- 14 

### C] label) AR. Organic volatile formation was neglig ible throughout the study (< 1 %).

For the dark controls , the average material balances ranged from 99.6 % to 103.4 % and from 101.6 % to 104.5 % of AR, respectively ([3- 14 

### C] and [5- 14 

### C] label). The extractable radioactivity decreased from an average of 99.5 % / 103.0 % of AR at day 0 to 67.4 % / 73.7 % by day 7, respectively. The NER formatio n was significant higher than built during irradiatio n with rates up to 30.9 % ([3- 14 

### C] label) and 27.5 % ([5- 14 

### C] label) AR at the end of the test. Organic volatile formation was negligible throughout the study (< 1 %).

In the irradiated test systems ([3- 14 

### C] label) the parent compound decreased from an average of 9 8.3 % of AR by DAT-0 to 36.5 % of AR at study termination (7 days). The major metabolite spirotetramatketohydroxy was detected at a maximum of 12.3 % of AR by DAT-7 . The minor degradates dimethylbenzoic acid, spirotetramat-glyoxylic amide and spi rotetramat-enol were formed at a maximum of 4.8 %, 1.1 % and 5.3 % of AR at study termination (DAT-7), res pectively. Spirotetramat-enol-dimer was not detecte d in comparison to the dark control ([3- 14 

### C] label).

In the irradiated test systems ([5- 14 

### C] label) the parent compound decreased from an average of 1 01.4 % of AR by DAT-0 to 30.8 % of AR at the end of the st udy (7 days). In addition the spirotetramat-ketohydroxy (max. 20.9 % of AR, mean of replicates, DAT-2) two further major metabolites were detected. Spirotetramat-enol reached a maximum of 10.1 % of AR at day 1 (mean o f replicates). Metabolite 4- methoxy-cyclohexanone (only detected in the [5- 14 

### C] label samples) reached a maximum of 10.0 % of AR (mean of replicates) by DAT-2 and then decreased to 6.0 % of AR (mean of replicates) at study terminat ion. The minor metabolites identified were spirotetramat -enol-dimer and spirotetramat-glyoxylic amide, whic h were formed at a maximum of 1.9 % of AR (DAT-2, mea n of replicates) and 4.1 % of AR (DAT-2, mean of replicates), respectively. Dimethyl-benzoic acid wa s not formed in the [5- 14 

### C] label test system.

In the dark test systems ([3- 14 

### C] label), spirotetramat decreased from 98.3 % (DAT-0, mean of replicates) to 9.4 % of AR at the end of the study. The two maj or metabolites identified were spirotetramat-enol and spirotetramat

- ketohydroxy with maximal amounts of 19.8 % of AR (DAT-2, mean of replicates) and 33.0 % of AR (DAT-7, mean of replicates), respectively. Mi nor transformation products, spirotetramat-enol-dim er and spirotetramat-glyoxylic amide, reached a maximum of 5.4 % of AR (DAT-4, mean of replicates) and 2.1 % of AR (DAT-2/3, mean of replicates), respectively.

In dark [5- 14 

### C] label test systems the picture was very similar. The two major transf ormation products, spirotetramat-enol and spirotetramat-ketohydroxy , were formed at a maximum of 16.6 % of AR (DAT-4, mean of replicates) and 33.9 % of AR (DAT-7, mean o f replicates), respectively. The minor degradates identified were spirotetramat-enol-dimer 1 and spir otetramat-glyoxylic amide with max. amounts of 8.6 % of AR (DAT-3, mean of replicates) and 3.1 % of AR (DAT -1, mean of replicates), respectively. No dimethylbenzoic acid was detected (neither in [3- 14 

### C] nor in [5- 14 

### C] label test system).

The experimental DT 50 

### values of spirotetramat in irradiated samples ([3- 14 

### C] and [5- 14 

### C] label) were 5.0 days and 2.4 days, respectively. In the dark controls th e experimental DT 50 

### of spirotetramat was 1.2 days ([3- 14 

### C] label) and 0.6 days ([5- 14 

### C] label). It can be concluded that the degradation of spirotetramat was approx. 4

### – 45 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

times faster in the dark systems than in the irradi ated systems. It was not possible to calculate the phototransformation rate constants, obtained by subtract ion ( k irradiated 

### – k

dark 

### ), because it resulted in negative
values.

Supplementary experiment: On dry irradiated ‘Molino’ soil samples, degradatio n of spirotetramat was significantly slower than on moist irradiated samples (based on 7 DAT findings). Howev er, the pattern of degradates was similar to the mo ist soil samples. Almost no degradation of spirotetrama t occurred in the dried soil samples kept under dar k
conditions.

On moist ‘Laacherhof AIII’ soil the parent compound showed a significant faster degradation than on mo ist ‘Molino’ soil based on 7-DAT findings. Formation of degradates under irradiated and dark conditions wa s similar to the moist ‘Molino’ soil. However, the ph otolytic metabolites dimethyl-benzylic acid and spirotetramat-glyoxylic amide were found at distinc t higher levels with 21.8 % of AR and 7.7 % of AR b y 7 DAT, respectively.

Table B.8.1.1.3-2: Distribution and recovery of rad ioactivity [% of AR] after application of spirotetramat ([3- 14 

### C] label, 288 g ai ha -1 

### ) on moist/dry ‘Molino’ soil surface (mean of dupli cate samples).

Conditions DAT Total extracted a 14 CO 2 Org. volatiles NER Recovery

0 99.5 na na 0.1 99.6
0.2 97.7 < 0.1 < 0.1 3.0 100.7

1 96.2 0.1 < 0.1 5.3 101.7
2 93.3 0.3 < 0.1 6.9 100.6
3 88.8 1.0 < 0.1 8.0 97.8

4 87.4 2.1 < 0.1 8.4 97.9 Irradiated (Moist soil)

7 81.7 3.8 < 0.1 9.6 95.2 Dry soil b 7 95.3 3.3 0.0 3.0 101.6

0 99.5 na na 0.1 99.6
0.2 96.7 < 0.1 < 0.1 6.7 103.4

1 80.7 0.3 < 0.1 21.8 102.8
2 73.9 0.5 < 0.1 27.8 102.1
3 68.0 1.0 <0.1 32.8 101.8

4 70.2 1.0 < 0.1 30.9 102.1 Dark (Moist soil)

7 67.4 1.6 < 0.1 30.9 99.9 Dry soil b 7 103.5 0.0 0.0 1.3 104.8 na denotes not applicable

a Including the final acetonitrile extract that was not further considered for analyses due to its low RA content
b Supplemental experiment (only 0 and 7 DAT sampling )

### – 46 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.1.3-3: Distribution and recovery of rad ioactivity [% of AR] after application of spirotetramat ([5- 14 

### C] label, 288 g ai ha -1 

### ) on moist/dry ‘Molino’ soil surface (mean of dupli cate samples).

Conditions DAT Total extracted a 14 CO 2 Org. volatiles NER Recovery

0 103.0 na na 0.1 103.1
0.2 99.5 < 0.1 < 0.1 5.2 104.7

1 95.2 0.6 < 0.1 7.4 103.2
2 87.6 2.3 < 0.1 9.3 99.2
3 86.5 3.2 < 0.1 9.8 99.5

4 81.4 5.3 < 0.1 10.9 97.6 Irradiated (Moist soil)

7 76.5 7.3 < 0.1 12.1 95.9 Dry soil b 7 94.4 1.8 0.0 2.4 98.6

0 103.0 na na 0.1 103.1
0.2 95.1 < 0.1 < 0.1 9.1 104.2

1 76.6 0.3 < 0.1 25.8 102.6
2 73.7 0.7 < 0.1 27.3 101.6
3 74.1 0.9 < 0.1 27.9 103.0

4 75.5 1.0 < 0.1 28.0 104.5 Dark (Moist soil)

7 73.7 1.4 < 0.1 27.5 102.7 Dry soil b 7 102.8 0.0 0.0 1.4 104.2 na denotes not applicable

a Including the final acetonitrile extract that was not further considered for analyses due to its low RA content
b Supplemental experiment (only 0 and 7 DAT sampling )



### Table B.8.1.1.3-4: Distribution and recovery of rad ioactivity [% of AR] after application of spirotetramat, both labels, 288 g ai ha -1 

### ) on moist ‘Laacher Hof AIII’ soil surface (mean of duplicate samples, supplemental experiment, only 0 and 7 DAT samplings).

Conditions Label DAT Total extracted 14 CO 2 Org. volatiles NER Recovery

[3- 14 C] 7 80.7 3.5 0.0 13.2 97.4 Irradiated [5- 14 C] 7 67.4 9.6 0.0 14.8 91.8

[3- 14 C] 7 68.4 3.5 0.0 26.9 98.9 Dark [5- 14 C] 7 70.9 2.0 0.0 25.7 98.6– 47 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.1.3-5: Characterization of extractableradioactivity [% of AR] after application of [3-14C] labelled spirotetramat(288 g ai ha-1) on moist/dry‘Molino’ soil surface (individual replicates).SpirotetramatSpiro-tetramat-enolSpiro-tetramat-enol-dimerDimethyl-benzoicacidSpiro-tetramat-glyoxylic-amideSpiro-tetramat-keto-hydroxyROI 3 ROI 8 ROI 9 ROI 10 ROI 11Unidenti-fieddiffuseRAConditions DATA B A B A B A B A B A B A B A B A B A B A B A B098.1 98.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.00.294.2 95.1 1.4 1.5 0.0 0.0 0.0 0.0 0.0 0.0 1.3 1.3 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0187.0 81.4 2.05.60.0 0.0 0.0 2.7 0.0 0.0 6.4 5.1 0.0 0.0 0.0 0.0 0.00.0 0.0 1.3 0.0 0.0 0.0 0.0273.3 76.4 3.4 1.9 0.0 0.0 3.5 1.9 0.0 0.0 8.3 9.3 0.0 0.0 0.0 2.5 0.0 0.0 2.9 2.3 0.0 0.0 0.0 0.0369.7 67.8 0.0 3.1 0.0 0.03.83.1 0.01.98.3 8.5 0.0 0.0 0.0 0.0 0.0 0.0 2.5 2.3 0.0 0.0 3.52.3453.7 53.8 1.7 2.2 0.0 0.03.84.2 1.4 0.012.910.0 0.0 0.04.6 2.70.0 0.0 2.8 3.7 3.04.93.6 4.5Irradiated(Moist soil)733.0 39.95.65.0 0.0 0.0 3.65.9 2.20.010.913.80.0 0.0 0.0 0.02.6 5.3 4.0 4.0 5.03.511.6 5.6Dry soilb777.0 ± 2.3 0.0 ± 0.0 nd 1.8 ± 0.2 1.6 ± 0.1 7.3 ± 0.1 nd nd nd 0.7 ± 0.7 1.7 ± 0.1 0.8 ± 0.8098.1 98.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.00.292.8 92.0 1.4 2.4 0.0 0.0 0.0 0.0 0.0 0.0 2.0 2.2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0148.6 57.815.4 9.2 2.6 1.8 0.0 0.0 1.5 0.011.810.8 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0226.2 27.319.819.93.7 3.0 0.0 0.01.92.319.018.9 0.0 0.01.00.0 0.0 0.0 1.1 1.4 0.0 0.0 0.0 0.0319.0 17.117.619.0 4.1 4.1 0.0 0.0 1.42.823.122.0 0.7 0.0 0.0 0.0 0.0 0.0 1.5 2.2 0.0 0.0 0.0 0.0413.5 14.515.815.85.1 5.80.0 0.0 1.5 0.028.730.4 1.7 0.0 0.0 0.0 0.0 0.0 2.8 2.2 0.0 0.0 0.0 0.0Dark(Moist soil)710.1 8.813.514.9 5.0 4.2 0.0 0.0 0.0 0.032.133.8 2.4 1.40.0 0.0 0.0 0.03.0 2.90.0 0.0 0.0 0.0Dry soilb798.0 ± 1.7 0.4 ± 0.4 nd 0.0 ± 0.0 0.6 ± 0.6 4.4 ± 0.4 nd nd nd 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0bSupplemental experiment (mean ± STD)Table B.8.1.1.3-6: Characterization of extractableradioactivity [% of AR] after application of [3-14C] labelled spirotetramat(288 g ai ha-1) on moist‘Laacherhof AIII’ soil surface (mean ± STD, supplemental experiment).Conditions DATSpiro-tetramatSpiro-tetramat-enolSpiro-tetramat-enol-dimerDimethyl-benzoicacidSpiro-tetramat-glyoxylic-amideSpiro-tetramat-keto-hydroxyROI 1 ROI 3 ROI 9 ROI 10 ROI 11 ROI 13Unidenti-fieddiffuseRAIrradiated 711.3 ± 2.5 1.2 ± 1.2 4.2 ± 0.521.8 ± 2.0 7.7 ± 1.724.7 ± 4.3 0.0 ± 0.0 0.6 ± 0.6 1.6 ± 1.6 1.7 ± 1.73.2 ± 0.6 0.0 ± 0.0 1.7 ± 0.3Dark 74.0 ± 0.3 9.1 ± 0.614.4 ± 0.0 0.0 ± 0.0 0.0 ± 0.026.9 ± 1.6 5.2 ± 0.0 1.3 ± 1.3 0.0 ± 0.0 0.0 ± 0.00.0 ± 0.0 3.5 ± 0.1 2.2 ± 0.6– 48 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.1.3-7: Characterization of extractableradioactivity [% of AR] after application of [5-14C] labelled spirotetramat(288 g ai ha-1) on moist/dry‘Molino’ soil surface (individual replicates).SpirotetramatSpirotetramatenolSpiro-tetramat-enol-dimerSpiro-tetramat-glyoxylicamideSpiro-tetramat-keto-hydroxy4-Methoxy-cyclo-hexa-noneROI 1 ROI 11 ROI 13 ROI 15 ROI 16 ROI 17 ROI 18Unidenti-fieddiffuseRAConditionsDATA B A B A B A B A B A B A B A B A B A B A B A B A BA B0100.7 102.2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.00.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.00.282.4 81.8 9.410.60.0 0.0 0.0 1.4 5.9 6.1 0.0 0.0 0.0 0.0 0.0 0.0 0.00.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0151.8 62.79.510.71.71.5 3.4 2.519.014.17.8 4.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.00.0 0.0 0.0 0.0 0.0238.5 35.8 7.0 9.5 1.42.3 4.4 3.919.322.510.79.3 0.0 0.0 1.5 0.0 0.0 0.0 0.0 1.5 2.2 2.8 0.0 0.00.0 0.0 0.0 0.0338.9 32.9 8.9 6.8 0.0 0.0 3.6 3.416.822.36.211.40.0 0.02.60.0 0.0 0.0 2.2 2.4 2.3 3.3 0.0 0.0 3.3 3.9 0.0 0.0431.3 35.1 4.2 2.6 0.0 0.0 1.9 2.519.418.98.7 7.5 0.0 0.0 0.0 0.0 0.0 0.0 2.84.2 4.3 6.90.0 0.05.0 4.41.1 0.0Irradiated(Moistsoil)729.7 31.8 3.1 4.9 0.0 1.6 3.0 3.818.215.16.4 5.7 0.0 0.0 0.0 0.0 0.0 0.04.42.1 3.8 2.01.90.0 3.7 4.22.1 3.4Dryb776.9 ± 3.6 1.2 ± 0.4 nd 1.6 ± 0.2 5.6 ± 0.5 5.0 ± 0.1 nd nd nd nd nd nd nd 2.4 ± 1.20100.7 102.2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.00.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.00.277.2 75.910.711.80.8 1.3 0.0 0.0 5.0 5.5 0.0 0.0 0.0 0.0 0.0 0.0 0.00.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0122.8 25.914.513.96.9 6.0 3.2 3.026.123.60.0 0.0 2.1 2.7 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.00.0 0.0 0.0 0.0 0.0216.7 13.013.715.67.4 8.23.32.229.828.70.0 0.0 1.93.90.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.00.0310.8 9.715.315.88.68.62.02.428.731.50.0 0.0 2.4 2.7 0.0 0.0 1.5 2.5 0.0 0.0 0.0 0.0 0.00.0 3.3 0.0 0.0 0.048.6 8.516.117.06.3 6.7 2.12.432.432.80.0 0.0 2.6 2.6 0.0 0.0 1.8 2.3 0.0 0.0 0.0 0.0 0.00.0 2.9 2.8 0.0 0.0Dark(Moistsoil)78.6 5.812.312.78.77.2 0.0 2.034.033.80.0 0.02.82.4 0.0 0.02.8 4.70.0 0.0 0.0 0.0 0.0 0.03.4 3.20.0 0.0Dryb796.4 ± 0.0 1.1 ± 0.3 nd 1.3 ± 0.0 3.5 ± 0.2 0.0 ± 0.0 nd nd nd nd nd nd nd 0.3 ± 0.3bSupplemental experiment (mean ± STD)Table B.8.1.1.3-8: Characterization of extractableradioactivity [% of AR] after application of [5-14C] labelled spirotetramat(288 g ai ha-1) on moist‘Laacherhof AIII’ soil surface (mean ± STD, supplemental experiment).ConditionsDATSpirotetramatSpiro-tetramat-enolSpiro-tetramat-enol-dimerSpiro-tetramat-glyoxylicamideSpiro-tetramat-keto-hydroxy4-Methoxycyclo-hexa-noneROI 1 ROI 3 ROI 13 ROI 15 ROI 16 ROI 18 ROI 20Unidenti-fieddiffuseRAIrrad. 75.0 ± 1.8 2.6 ± 0.2 3.2 ± 0.4 5.2 ± 3.922.8 ± 4.6 3.5 ± 1.5 0.7 ± 0.7 1.2 ± 1.2 1.5 ± 1.52.7 ± 0.1 2.0 ± 0.5 4.9 ± 0.3 1.9 ± 0.0 9.1 ± 0.9Dark 74.1 ± 0.4 9.5 ± 0.710.4 ± 2.3 0.0 ± 0.020.1 ± 0.2 0.0 ± 0.0 3.5 ± 0.7 1.9 ± 0.2 4.5 ± 0.32.5 ± 0.5 0.0 ± 0.0 2.9 ± 0.9 1.5 ± 0.0 8.1 ± 1.1

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.1.3-9: Calculated DT 50 

### and DT 90 

### of spirotetramat on ‘Molino’ soil surface under ir radiated and dark conditions using SFO kinetics ([3- 14 

### C] and [5- 14 

### C] label).

Soil Label Test system DT 50 [days] DT 90 [days] R 2 chi 2 error [%] a Kinetics

Irradiated 5.0 16.6 0.991 2.2 SFO [3- 14 C] Dark 1.2 3.9 0.986 7.4 SFO

Irradiated 2.4 8.1 0.830 15.6 SFO ‘Molino’ [5- 14 C] Dark 0.6 1.9 0.973 13.4 SFO

a calculated by the reviewer



### Conclusion: It was not possible to assess the rate of photodegr adation of spirotetramat on moist soil because most degradation was due to microbial activity (which wa s more active under dark conditions). Under dry conditions microbial activity was reduced and spiro tetramat photodegraded, but this (supplemental) stu dy had too few data points to calculate a half-life. F our major (> 10 % of AR) transformation products we re found in the irradiated soil samples on moist soils , i.e. spirotetramat-enol

, spirotetramat-ketohydroxy

, 4- methoxy-cyclohexanone (only [5- 14 

### C] label) and dimethyl-benzoic acid (only [3- 14 

### C] label). Minor transformation products determined were spirotetram at-glyoxylic amide and spirotetramat-enol dimer.

Reviewer comments:

• For the photolytic metabolite 4-methoxy-cyclohexan one a separate aerobic degradation rate study was conducted (refer to B.8.1.2.3, aerobic degradat ion rate of relevant metabolites).

• Owing to the fact, that the irradiated test system was likely to be strongly affected by decreased biological activity, calculation of DT 50 

### values under environmental conditions is not consi dered to
result in reliable values.

• Metabolite dimethyl-benzoic acid was found > 10 % of AR in the supplemental experim ent (only 7 DAT sampling available) with ‘Laacherhof AIII’ so il with 21.8 % of AR. Maximum observed levels of dimethyl-benzoic acid in the main study (kinetic) p art (‘Molino’ soil) were 4.8 % of AR (mean). Both test systems suffered from low microbial activity o wing to the experimental conditions. In the outdoor metabolism study (B.8.1.1.1; Babszinski, 2006a), co nducted on the same two soils to simulate real environmental conditions including formulated appli cation and natural sunlight, maximum levels of dimethy-benzoic acid found were 3.3 % of AR (decrea sing thereafter) indicating that this metabolite is unlikely to occur at relevant amounts in viable soi ls. Therefore, dimethyl-benzoic acid is not considered a relevant soil photolysis metabolite by the reviewers.

• Reviewers from the US-EPA and from the PMRA requir ed further studies using sterile soils to minimize microbial degradation. On request the noti fier (BCS) provided further information: BCS: ‘The study was conducted according to establis hed guidelines as required with viable soil (EPA § 161-3 and OECD Draft TG dated Feb 2002). As acknowledged by the reviewers, considering all the results on the behaviour of spirotetramat in soils, BCS regards the phototransformation of spirotetramat of no greater impor tance compared to the dark biodegradation. Thus, by using a sterile soil, not much information to real-world exposure would be added. The real world situation is better described by the res ults of an outdoor soil metabolism study, regarded as an adequate supportive study, better th an a highly artificial study with sterile soil. In general the main objective of the photo-transformat ion study on soil surface is not to provide experimental DT 50 

### and DT

90 

### (it is not regarded as a rate of degradation study ), but to elaborate
what further metabolites should be considered for T FD studies and for exposure risk assessments in soil. It is clearly regarded as a ro ute of degradation study.’ However, to overcome several uncertainties related to the photodegradation rate of spirotetramat and formation of photolysis metabolites (see comments a bove), the notifier (BCS) is currently conducting an additional soil photolysis study on sterile soil . This study is expected for submission by end of
2008.

### – 50 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.1.4 Summary on route of degradation in soil ( laboratory)

The route of degradation of spirotetramat was estab lished on 3 EU soils and one US soil in two studies using either spirotetramat or spirotetramat-enol (which i s rapidly released from spirotetramat) as parent. T he latter study (using alkaline extraction) was conducted to overcome metabolite instability observed in the spirotetramat study during the acidic extraction pr ocedure. In this respect, only the degradation stud y using spirotetramat-enol as parent is considered relevant for evaluation of maximum occurrence (and degradat ion rates) of metabolites. However, despite the analyti cal problems, both studies are consistent in their overall degradation pathway of spirotetramat in aerobic soi l.

Aerobic laboratory soil degradation studies were co nducted under static conditions (no flow through systems). Both test systems (static and flow throug h) are options outlined in the test guidelines. Fro m the notifier’s long-term experience with the static sys tems the validity of that test system is fully prov en indicated by evolution of carbon dioxide formed till the end of the aerobic studies. Furthermore, the terminal b iomass determinations indicate a viable test system throug h the entire test duration.

Under standard aerobic conditions (20 °C), the metabolism of spirotetramat ([3- 14 

### C] label) was shown to proceed via three pathways which all start with the hydrolytic cleavage of the spirotetramat ester int o the spirotetramat-enol (Figure 2.5.2-1). Following the major degradation pathway, spirotetramat-enol is ra pidly oxidized at the benzylic carbon position into spiro tetramat-ketohydroxy which is hydrolytically opened into spirotetramat-MA-amide and finally mineralized into CO 2 

### . In the second (minor) pathway demethylation of spirotetramat-enol into spirotetramat-desmethyl-eno l and oxidation of this degradate into spirotetrama t-oxoenol occurred. In a third degradation pathway spiro tetramat-enol-dimer 1 and spirotetramat-enol-dimer 2 were pretended to be built through oxidative enol d imerization. However, this pathway is regarded as n on relevant under environmental conditions, because th e formation of dimers is considered an artificial p rocess mainly caused by the hot-spot application in this l aboratory tests.

As mentioned above, no final conclusion onto the ma ximum occurrence (and degradation rates) of metabolites of spirotetramat could be drawn from th e parent degradation study. In the second degradati on study, using spirotetramat-enol as parent ([3- 14 

### C] and [5- 14 

### C] label) and the same test soils as used in the parent degradation study, spirotetramat-ketohydroxy was observed at maximum amounts of 24.0 % of AR (by 1 DAT), spirotetramat-MA-amide was observed at maximum amounts of 5.2 % of AR by 7 DAT (only one sample > 5 % of AR). No other metabolite was observ ed > 5 % of AR in this study. Spirotetramat-oxoketohydroxy, a metabolite previously not found, was a very minor compound and was not quantifiable. Spirotetramat-desmethyl-enol was observed at maximu m amounts of 1.8 % of AR by 4 DAT. Since the maximum occurrence of spirotetramat-enol can not be adequately derived from the parent study, maximum occurrence of spirotetramat-enol was set to 100.0 % of AR in all risk assessments for conservative rea sons.

Formation of CO 2 

### in the spirotetramat degradation study was in a ra nge of 9.7 – 19.4 % of AR by 50 DAT (i.e. the study termination in case of the EU soils ), in the US soil, CO 2 

### further increased from 9.7 % of AR by 50 DAT to 15.3 % of AR by study termination (360 DA T). No volatile organics could be detected. The soi l with the highest content of microbial biomass showe d the highest mineralization rate. Formation of CO 2 

### in the spirotetramat-enol degradation study was in a r ange of 16.7 – 43.0 % of AR by study termination (1 19 DAT). Significant higher 14 

### CO 2 

### amounts were released from spirotetramat-enol labe lled in the [3- 14 

### C] position. No volatile organics could be detected.

In the spirotetramat study, the portion of non-extr actable radioactivity (NER) rapidly increased (by 1 – 3 DAT), reaching more or less a plateau concentration in the range of approx. 22 – 35 % of AR, hardly decreasing thereafter. A similar but even more pron ounced formation of NER was determined in the spirotetramat-enol study using a slightly alkaline extraction procedure. Directly after application (0 -DAT

### – 51 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

sampling) already 4.2 – 28.4 % of AR was bound to t he soil matrix. Amounts of NER rapidly increased further to reach a plateau concentration by 1 DAT a nd thereafter. Plateauing concentration was in a ra nge of 40 – 60 % of AR. No significant decrease of NER was found with time.

Further attempts were undertaken by the notifier to identify/characterize the pronounced NER fraction in both experiments by means of humic acid fractionation. A partitioning of the heterogeneous NER into humic a cid, fulvic acid and humin like fractions was observed, with the major portion belonging to the fulvic acid fraction and a significant portion bound to soil in the inso luble humin fraction.

During the evaluation, there was some discussion ab out the relevance of this extensive formation of NE R (or bound residues, BR) found in the aerobic lab degrad ation studies (especially, when spirotetramat-enol was used as parent). The general point is that the noti fier’s statement, that ‘ NER is immobile and not (readily)
bioavailable or bioaccessable’ , is not supported (by data). Reviewers from the PM RA believe that studies conducted on sterile and non-sterile soil may help to clarify this point. The EU project ‘FOOTPRINT’ 1 

### points
out that ‘ there is no agreement amongst EU Member States on h ow NER should be treated. The possible key to this question could be NER ‘bioavailability’ (Craven, 2000) 2 

### . The current approach which is most often adopted is to treat soil NER in the same way as per sistent compounds (Craven & Hoy, 2005) 3 

### . Thus, for these authors, the full range of environmental fate , long term effects and crop residue tests required to identify any risks posed by NER are brought into pl ay. This position is however not consensual. The Scientific Committee on Plants (SCP) provided a vie w on the issue in 1999 after analysis of the availa ble
literature 4 

### . The SCP noted that small fractions of NER may be released by a variety of processes and become potentially bioavailable. The opinion of the SCP (1999) is that the fractions of NER which are released are low and any concern over their ecotoxi cology or effect on succeeding crops should have be en addressed during studies required under Annex II an d III for the active substance and relevant metabol ites. Thus, for the SCP, the fractions released from NER have little significance from a regulatory viewpoin t.’

In an additional outdoor study, the degradation of [3- 14 

### C] labelled spirotetramat (spirotetramat formulated as an OD 100) was investigated under more realistic en vironmental conditions including natural sunlight. In comparison to the laboratory degradation study, bes ide the already mentioned metabolites three new metabolites were identified within this study: spir otetramat-glyoxylic amide (maximum 2.3 % of AR by 7 DAT), dimethyl-benzoic acid (3.3 % of AR by 7 DAT) and spirotetramat-ketohydroxy-carboxy (1.3 % of AR)

. Further, it has to be concluded that the formation of the spirotetramat-enol dimers within the outdoor study with another application technique (broadcast appli cation) was about ≤ 1.5 % of AR. Maximum formation of
NER was distinct lower than observed in the laborat ory studies with 16.8 % of AR by 63 DAT decreasing thereafter indicated an enhanced overall degradatio n of the parent and its metabolites in soil under r eal outdoor conditions in comparison to the laboratory degradation studies.

1 FOOTPRINT: Creating tools for pesticide risk assess ment and management in Europe. A project funded by th e European Commission under the 6 th Framework Programme (http://www.eu-footprint.org)
2 Craven A. (2000) Bound residues of organic compounds in the soil: the significance of pesticide persist ence in soil and water: a European regulatory view. Environmental Pollution , 108: 15-18.
3 Craven A. & Hoy S. (2005) Pesticide persistence and b ound residues in soil – regulatory significance. Environmental Pollution , 133: 5-9.
4 SCP Opinion on Draft GD on persistence in soil (DG i Y-9188/IV/97-Rev. 5, 1998), dated 1999

### – 52 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

NH O

O O O

CH 3

CH 3 H 3 C

NH O

O CH 3

CH 3 OH

CO 2 N H

O CH 3

CH 3 OH COOH NH O H

O CH 3

CH 3 OCH 3

NH O H

O CH 3

CH 3 OH

NH O H

O CH 3

CH 3 

### O NH

O CH 3

CH 3

NH O

O CH 3

CH 3 OH OH

NH O

O CH 3

CH 3 OH 

### O

N H

O CH 3

CH 3 O COOH CH 3

CH 3 O COOH OCH 3

NH O

O CH 3

CH 3 O OCH 3

OCH 3

OCH 3

NH O H

O CH 3

CH 3

OCH 3

OCH 3

OCH 3 OCH 3 NH O

O R

R1 OH OCH 3 NH

O H

O CH 3

CH 3 OH OCH 3 BYI08330

Enol

Ketohydroxy

Bound residues (- 2 H)

Enol-dimer 1 2

MA-amide Desmethyl-enol

Oxo-enol Hydrolysis

Oxidation x 2

Hydrolysis Desmethylation

Oxidation Enol-dimer 2

Desmethylation

ketohydroxyDesmethylOxo-ketohydroxy Oxidation 

## *

Benzoic
acid 

## ** Methoxy cyclohexanone Molecule

cleavage

Glyoxylic amide 

## **

Oxidation



## *** x 2 (- 2 H)

Oxidation

Oxidation

Ketohydroxycarboxy Oxidation

R,R 1 = -CH 3 , -COOH Oxidation

Oxidation

cleavage Molecule Dihydroxy

Oxidation Hydratization

*: postulated intermediate; ** and *** were identi fied in studies including light (dependent on radio label used)



### Figure B.8.1.1.4-1: Proposed metabolic pathway for spirotetramat (BYI08330) in soil.

### – 53 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Under anaerobic conditions the degradation pathway is almost identical to the degradation pathway obtained in aerobic soil. It was shown that spirote tramat declines quickly in a subsequently flooded a naerobic
soil situation. The detected metabolites were almos t identical to those formed upon the aerobic transformation in the aerobic metabolism study, exc ept of the minor metabolite spirotetramat-di-hydrox y. But spirotetramat-di-hydroxy was formed in the aerobic aquatic metabolism study. Therefore it can be exclu ded that this metabolite will be specially formed under anaerobic conditions.

It was not possible to assess the rate of photodegradation of spirotetramat on moist soil because most degradation was due to microbial activity (which wa s more active under dark conditions). Under dry conditions microbial activity was reduced and spiro tetramat photodegraded, but the (supplemental) stud y had too few data points to calculate a half-life. I n general, phototransformation on moist soil surfac es is not considered to significantly contribute to the overa ll dissipation of spirotetramat from the environmen t owing to the extremely rapid degradation under moist conditi ons. Four major transformation products (> 10 % of AR) were found in the irradiated soil samples, i.e. spi rotetramat-enol, spirotetramat-ketohydroxy, 4-metho xycyclohexanone (only observable using [5- 14 

### C] labelled parent) and dimethyl-benzoic acid (only observable using [3- 14 

### C] labelled parent). Minor transformation products determined were spirotetramat-glyoxylic amide and spirotetramat-enol dimer. 4-Methoxy-cyclohexano ne and dimethyl-benzoic acid were not found in dark samples indicating that these metabolites are photo lysis metabolites, only. Dimethyl-benzoic acid was shown not to exceed 3.3 % of AR in the outdoor study incl uding natural sunlight. Therefore, dimethyl-benzoic acid is not considered a relevant soil photolysis metabolit e. In case of 4-methoxy-cyclohexanone, which could not be determined in the outdoor study owing to the lab el used, a separate degradation study was conducted (refer to ‘rate of degradation’ section) in order t o derive appropriate degradation rates for this pho tolysis
metabolite in viable soils.

To overcome several uncertainties related to the ph otodegradation rate of spirotetramat and formation of photolysis metabolites (see comments above), the no tifier (BCS) is currently conducting an additional soil photolysis study on sterile soil . This study is expected for submission by end of 2 008.

Table B.8.1.1-10: Summary on maximum occurrence [% of AR] of metabolites of spirotetramat in soil degradation studies (numbers in brackets indicate d ay of maximum occurrence).

Compound Parent degradation study a Spirotetramat

-enol degradation study Outdoor

study Anaerobic

degradation study (entire system) Soil

photolysis study

Spirotetramat-enol 24.3 (3)

- b 7.8 (7) 54.6 (180) 10.1 (1) Spirotetramat-ketohydroxy 16.3 (1) 24.0 (1) 25.3 (14) 19.3 (1) 24.7 (7) Spirotetramat-MA-amide 6.4 (7) 5.2 (4) 6.2 (28) 7.2 (180) nd Spirotetramat-desmethyl-enol 3.7 (179) 2.0 (4) nd nd nd Spirotetramat-oxo-enol 1.2 (50) nd nd nd nd Spirotetramat-glyoxylic amide nd nd 2.3 (7) nd 7.7 ( 7) Spirotetramat-di-hydroxy nd nd nd 3.2 (32, 90) nd Dimethyl-benzoic acid c nd nd 3.3 (7) nd 21.8 (7) 4-Methoxy-cyclohexanone no no no no 10.0 (2) Spirotetramat-enol-dimer 1 d 12.7 (7) 5.0 (6) 1.5 (28) 7.0 (6) 4.2 (7) Spirotetramat-enol-dimer 2 d 9.2 (30) 3.6 (14) 0.9 (7) 4.6 (180) nd

no denotes not detected no denotes not observable (owing to the label used)

a Not considered relevant for evaluation of maximum o ccurrence of metabolites owing to analytical reason s b Set to 100.0 % for risk assessment, worst case ass essment

c Not considered relevant for risk assessment owing t o the fact, that dimethyl-benzoic acid was shown no t to exceed 3.3 % of AR in an representative outdoor study (B.8.1.1.1; Babczinski , 2006a) including natural sunlight (study conducte d with the same soils used in the laboratory soil photolysis study)
d Not considered relevant for risk assessment owing t o artifactual formation during hot-spot application (hardly observed under realistic outdoor and field conditions, see position paper Hel lpointner et al., 2007)

### – 54 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.2 Rate of degradation in soil (OECD Annex IIA , 7.2, IIIA, 9.1)

B.8.1.2.1 Aerobic degradation rate of the active su bstance at 20 °C (OECD Annex IIA, 7.2.1, IIIA,

9.1.1)

Data on the aerobic degradation rate of spirotetram at at 20 °C are available from one degradation rout e and rate study (Babczinski, 2005). In addition, the kin etic evaluation was described in a separate report, in which the methods and results of the respective kinetics calculations were described more in detail.

• Aerobic degradation in four soils (20 °C, [3- 14 

### C] label)

Full study details are given under B.8.1.1.1.

Reference: Degradation of BYI08330 in Four Soils Under Aerobic Conditions: Kinetic
Evaluation Author(s), year: Ramanarayanan, T., Roepke, B., 200 6 Study/report number: M-277096-01-2, MEF-05/249 Guideline(s): FOCUS guidelines (FOCUS, 2000 and FOC US, 2006) for kinetics modelling were
considered GLP: Calculations – not applicable Validity: Study considered acceptable

Material and methods: The degradation behaviour of [3- 14 

### C] labelled spirotetramat was studied in three Euro pean soils and one US soil under laboratory aerobic conditions at 20 °C ( B.8.1.1.1., Babczinski, 2005). A kinetic modelling analysis of residue data of spirotetramat was conducted usin g MatLab 7.0.4. (MatLab, 2005). The degradation kin etic calculations of the major metabolites were based on the spirotetramat-enol degradation study and were calculated in a separate study (Röpke, 2006a).

For moisture normalization the correction factor of each soil was estimated according to the recommendations of the FOCUS working group. Therefo re, FOCUS default values for each individual soil type for both, reference moisture content ( Θ ref 

### ) and study moisture content ( Θ)

 , were used (see Table B.8.1.2.1-1).

For characterization of the degradation behaviour o f spirotetramat single first order (SFO) and bi-pha sic kinetic (FOMC and DFOP) models were used. C 0 

### was calculated by the model.

The goodness of fit was assessed by visual inspecti on and an error criterion based on a chi-square ( 

### χ 2 

### ) test. A t-test was employed to identify the probability t hat a parameter is not significantly different from zero. The
coefficients of determination r 2 

### were also reported for comparative reasons. The ki netics modelling tool is built within MatLab (Vers. 7.0.4). The tool uses in ternal routines (Levenberg-Marquardt) of MATLAB to optimize the model parameters to fit a chosen kinet ics model to the metabolism study data. The objecti ve function used for the optimization of the parameter s was to minimize the sum of squares between the
calculated and observed time series data.

Findings: The results of the SFO calculation showed that spir otetramat degraded very quickly during the first da y to
less than 10 % of AR in all four soils. Afterwards the degradation slowed down. The visual assessment of the
measured and the observed data showed that the SFO kinetic model explains the degradation of

### – 55 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

spirotetramat very well during the initial periods of the study (less than 1 day) and underestimates t he concentrations during the later time points for all soils (especially for ‘Molino’ soil). Therefore bi -phasic models as FOMC and DFOP were used to improve the vi sual and statistical assessment of the fits (according to trigger evaluation).

Comparing the visual fits and the goodness of fit p arameters between the two chosen bi-phasic models i t can be concluded that the DFOP model showed a more suit able fit than the FOMC model. The estimated DT 50 

### values were very close to the values derived from t he SFO model fit, but the DT 90 

### values were somewhat higher.– 56 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.2.1-1: Normalization factors for soil moisture (fMoist).Moisture contentduring study[g 100 g-1]Moisture contentat pF2[g 100 g-1]cfMoistSoilSoil texture(USDA)Study moistureMWHC[g 100 g-1]aMoisture contentat pF 2.5b[g 100 g-1]ΘΘref7.0MoistfΘΘ=ref‘Laacherhof AXXa’ Sandy loam 50 % of MWHC 27 (15) 13.5 19 0.79‘Laacherhof AIII’ Silt loam 50 % of MWHC 32 (21) 16.0 26 0.71‘Hoefchen’ Silt 50 % of MWHC 31 (21) 15.5 27 0.67‘Molino’ Sandy loam 75 % of MC at pF2.5 (27) 15 11.3 19 0.69aMWHC for classified soils taken from the FOCUS report (FOCUS, 2000)bEstimates for classified soils taken from the FOCUS report (FOCUS, 2000)cEstimates for classified soils taken from the FOCUS report (FOCUS, 2000)

### – 57 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.2.1-2: Degradation rates of spirotetram at in aerobic lab studies (20 °C) based on SFO kinetics – corrected by the reviewer. a

Soil C 0

[% of AR] DT 50

[days] DT 90

[days] a χ 2 error [%] R 2 p of t-test DT 50 norm

[days]

‘Laacherhof AXXa’ 91.5 0.21 0.69 8.52 > 0.99 < 0.001 0.17
‘Laacherhof AIII’ 91.2 0.23 0.77 8.81 > 0.99 < 0.001 0.16
‘Hoefchen’ 93.2 0.08 0.28 9.50 > 0.99 < 0.001 0.05
‘Molino’ 94.5 0.33 1.09 21.79 0.99 < 0.001 0.23

a Wrong values were transferred from the result sheet s to the summarizing study report.



### Table B.8.1.2.1-3: Degradation rates of spirotetram at in aerobic lab studies (20 °C) based on FOMC kinetics – corrected by the reviewer. a

Soil C 0

[% of AR] DT 50

[days] DT 90

[days] DT 50recalc b

[days] χ 2 error

[%] R 2 p of t-test DT 50recalc

norm [days]

‘Laacherhof AXXa’ 91.8 0.25 1.04 b 0.31 6.51 b > 0.99 - 0.24
‘Laacherhof AIII’ 91.6 0.23 1.09 b 0.33 7.47 b > 0.99 - 0.23
‘Hoefchen’ 93.2 0.01 0.44 0.13 3.61 > 0.99 - 0.09
‘Molino’ 96.6 0.26 2.61 0.79 16.5 > 0.99 - 0.55

a Wrong values were transferred from the result sheet s to the summarizing study report.
b SFO-DT 50 recalculated from the FOMC-DT 90 divided by 3.32.



### Table B.8.1.2.1-4: Degradation rates of spirotetram at in aerobic lab studies (20 °C) based on DFOP kinetics

.

Soil C 0

[% of AR] DT 50

[days] DT 90

[days] DT 50recalc a

[days] χ 2 error

[%] R 2 p of t-test DT 50recalc

norm [days]

‘Laacherhof AXXa’ 91.7 0.24 0.89 0.27 1.37 > 0.99 < 0.001 0.21
‘Laacherhof AIII’ 91.5 0.26 0.97 0.29 0.81 > 0.99 < 0.001 0.21
‘Hoefchen’ 93.2 0.09 0.34 0.10 4.0 > 0.99 < 0.001 0 .07
‘Molino’ 95.5 0.30 1.26 0.38 6.12 > 0.99 < 0.001 0. 26

a SFO-DT 50 recalculated from the DFOP-DT 90 divided by 3.32.



### Conclusion: The DFOP kinetic model fitted the experimental data best in terms of visual assessment and the χ 2 

### scaled errors. However, SFO is the preferred model for der iving parameters to be used in environmental fate models. The visual assessment of the SFO plots indi cated an acceptable fit of the data to the SFO mode l for three of the four soils (‘Laacherhof AXXa’, ‘Laache rhof AIII’ and ‘Hoefchen’), the χ 2 

### scaled errors were below 10 % indicating good agreement between the modelled and measured data. Despite the fact, that the χ 2



### scaled error in case of the ‘Molino’ soil was estim ated to be 8 % higher than the indicator value sugg ested by the FOCUS group (i.e. 15 %, FOCUS, 2006), the SFO m odel was found to adequately represent all four
studied soils.

It can be concluded that spirotetramat degrades ver y quickly with DT 50norm 

### values < 1 day.

Reviewer comments:

• Normalization procedure was based on FOCUS default values for each individual soil type for both, reference moisture content ( Θ ref 

### ) and study moisture content ( Θ) . Although the study moisture content was measured and is therefore exactly known (except for soil Molino), the approach by the notifier is considered acceptable.

• The notifier stated normalized FOMC- and DFOP-DT 50 

### values in the study report. The reviewer is of the opinion that normalization of non-SFO kinetics is not advisable at all since processes which result in non-SFO kinetics behaviour (e.g. increasi ng adsorption with time, decreasing bioavailability

, etc.) do not necessarily follow common normalizatio n assumptions (moisture, temperature correction

### – 58 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

factors, etc.). Therefore, the reviewers based the normalization procedure on recalculated SFO-DT 50 

### values (i.e. FOMC-DT 90 

### or DFOP-DT 90 

### divided by 3.32), which is generally considered a conservative approach.

B.8.1.2.2 Aerobic degradation of the active substan ce at 10 °C (OECD Annex IIA, 7.2.2)

The rate of aerobic degradation of spirotetramat at 10 °C was calculated by the notifier using the Q 10 

### -value of 2.2 (see Tier 2, AII, 7.2.2): Multiplying the DT 50 

### values of 0.08 to 0.33 days (SFO kinetics) by a fa ctor of 2.2
will result in DT 50 

### values between 0.18 and 0.73 days.

### – 59 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.2.3 Aerobic degradation of relevant metabolit es at 20 °C (OECD Annex IIA, 7.2.3)

Data on the aerobic degradation rate of major spiro tetramat metabolites are available from one degrada tion route and rate study with spirotetramat-enol (B.8.1 .1.1). The kinetic evaluation was described in a se parate report, in which the methods and results of the res pective kinetics calculations were described more i n detail.

• Spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amide:
Aerobic degradation/metabolism in four soils (20 °C, [3- 14 

### C] and [5- 14 

### C]) label) - full study details are given under B.8.1.1.1 (Babczinski, 2006b)

• Methoxycyclohexanone (photolytic metabolite): Aerobic degradation in three soils (20 °C)

Reference: Kinetic Evaluation of Laboratory Soil Degradation S tudies of BYI08330-Enol, BYI08330-Ketohydroxy, BYI08330-MA-Amide Author(s), year: Roepke, B., 2006a Study/Report number: M-277178-03-1, MEF-06/199, ame nded 23.08.2007 Guideline(s): FOCUS guidelines (FOCUS, 2000 and FOC US, 2006) for kinetics modelling were
considered GLP: Calculations - not applicable Validity: Study considered acceptable

Material and methods: The aerobic soil degradation study with spirotetram at-enol (Babczinski, 2006b) was used to estimate ki netic parameters and half-lives of spirotetramat metaboli tes spirotetramat-enol, spirotetramat-ketohydroxy a nd spirotetramat-MA-amide. It was not possible to get adequate formation fractions from the parent degrad ation study since under acidic extraction conditions, whi ch was needed for spirotetramat but was not applica ble for spirotetramat-enol, spirotetramat-enol was found to be partly unstable. For this reason a separate stu dy with spirotetramat-enol was conducted.

In the course of the spirotetramat-enol degradation study spirotetramat-enol showed a biphasic kinetic behaviour, whereas in the first phase (during one d ay) more than 80 % of AR “has degraded”. In the sec ond phase a much slower degradation was shown (during 1 18 days from 10.7 – 17.2 % of AR to 2.7 – 6.1 % of AR, mean of both labels).

The degradate spirotetramat-ketohydroxy was built u p to a maximum of 24.0 % of AR at DAT-1 (mean of both labels) and spirotetramat-MA-amide reached a m aximum of 5.2 % of AR (mean of both labels).

For moisture normalization procedure the appropriat e moisture normalization factor was calculated:– 60 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.2.3-1: Normalization factors for soil moisture (fMoist).Moisture contentduring study[g 100 g-1]Moisture contentat pF2[g 100 g-1]cfMoistSoilSoil texture(USDA)Study moistureMWHC[g 100 g-1]aMoisture contentat pF 2.5b[g 100 g-1]ΘΘref7.0MoistfΘΘ=ref‘Molino’ Sandy loam 80 % of MC at pF2.5 (27) 15 12.0 19 0.72‘Laacherhof AXXa’ Sandy loam 60 % of MWHC 27 (15) 16.2 19 0.89‘Laacherhof AIII’ Silt loam 60 % of MWHC 32 (21) 19.2 26 0.81‘Hoefchen’ Silt 60 % of MWHC 31 (21) 18.6 27 0.77aMWHC for classified soils taken from the FOCUS report (FOCUS, 2000)bEstimates for classified soils taken from the FOCUS report (FOCUS, 2000)cEstimates for classified soils taken from the FOCUS report (FOCUS, 2000)

### – 61 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

All values (both labels singular) measured were use d for kinetic evaluation except of one outlier iden tified for spirotetramat-enol in soil AIII at DAT-4 (both labe ls). Samples with non-detects (nd) were set to 0.5 LOD. Samples marked with < LOQ were also set to 0.5 (LOQ + LOD).

During sampling processing (starting with an aqueou s extraction) spirotetramat-enol degradation starte d and directly after application oxidation yielded to spi rotetramat-ketohydroxy in amounts of up to 4.9 and 3.3 % of AR ([3- 14 

### C] and [5- 14 

### C] label). It was shown in stability tests that 79. 8 % to 90.1 % of HPLC-injected RA for the [3- 14 

### C] labelled test item and 85.7 % to 91.5 % for the [5- 14 

### C] labelled test item were obtained during aqueous DAT-0 extraction. To consider the ongoing d egradation directly at application and extraction procedure an additional DAT-0 was introduced for mo delling purposes assuming 100 % of AR for spirotetramat-enol and 0 % of AR for spirotetramat- ketohydroxy and spirotetramat-Ma-amide. Consequentl y, this approach led to four DAT-0 values for the pare nt and the metabolites, i.e. two measured ones and two fixed to 100.0 % of AR (parent) and 0.0 % of AR (me tabolites), which were taken into account for model ling.

Beside of single first order models (SFO, single fi rst order; FOMC, first order multi compartment; and DFOP, double first order in parallel) a single first orde r reversible binding model (SFO-RB, a bi-phasic mod el consisting of two compartments, an unprotected comp artment where application and degradation occurs an d a protected compartment assuming no degradation, FO CUS, 2006) was considered to most adequately describe the degradation behaviour of metabolite sp irotetramat-enol. The metabolites spirotetramatketohydroxy and spirotetramat-MA-amide were assumed to follow SFO kinetics, only. No weighting of the data was performed. As default value, free fitting of the initial parent amount and fixing amount to z ero for metabolites and equal weighting was used.

As the SFO-RB model is conceptually similar to the kinetic desorption model implemented in PEARL, required sorption and degradation parameter can be directly taken from the SFO-RB parameters:

Pearl SFO-RB
K t 

### k 1 

### k d 

### k 3 

### φ k 2 

### /k 3



### The parameter φ is not a PEARL input parameter per se , the corresponding input parameter is f ne 

### . As outlined in FOCUS (2006), the f ne 

### value can be deduced from φ as follows:

EQ L EQ L

ne 

### K K f

, ,



### ⋅ ⋅ +

= ρ ρ θ φ ,

whereby θ is the volumetric water content of the soil [g g -1 

### ] and ρ is the bulk density [g cm -3 

### ] (data available from the laboratory degradation rate study, Babczin ski, 2006b). In case of spirotetramat-enol, the K L,EQ 

### [L kg -1 

### ] is related to the K d 

### value obtained in the column leaching study using spirotetramat-enol as parent (Babczinski, 2006c).

The proposed metabolic route of spirotetramat-enol in soil was converted to a four-compartment model (assuming SFO, FOMC or DFOP kinetics for the degrad ation of spirotetramat-enol and SFO kinetics for th e degradation of spirotetramat-ketohydroxy and spirot etramat-MA-amide) and a five-compartment model (assuming SFO-RB kinetic model for spirotetramat-en ol and SFO kinetics for spirotetramat-ketohydroxy a nd spirotetramat-MA-amide).

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Figure B.8.1.2.3-1: Four-compartment model for fitt ing spirotetramat-enol (following SFO, FOMC or DFOP kinetics), spirotetramat-ketohydroxy (always S FO kinetics) and spirotetramat-MA-amide (always SFO kinetics) residues in soil.

Figure B.8.1.2.3-2: Five-compartment model for fitt ing spirotetramat-enol (following SFO-RB kinetics), spirotetramat-ketohydroxy (SFO) and spir otetramat-MA-amide (SFO) residues in soil.

The appropriate kinetic model was identified by fol lowing the approach recommended by FOCUS (2006) and was based on visual inspection and the scaled error ε = σ / y.

The goodness of fit was assessed by visual inspecti on and an error criterion based on a chi-square ( 

### χ 2 

### ) test. A t-test was employed to identify the probability t hat a parameter is not significantly different from zero. The
coefficients of determination r 2 

### were also reported for comparative reasons. The ki netics modelling tool is built within MatLab (Vers. 7.0.4). The tool uses in ternal routines (Levenberg-Marquardt) of MatLab to optimize the model parameters to fit a chosen kinet ics model to the metabolism study data.

Findings: In a first step, considering the evaluation of mode lling and trigger endpoints, SFO kinetics was fitte d to all unmodified data. The visual inspection and the scal ed error ε with a range from 20.9 – 37.4 % for spirotetramat-enol, 24.2 – 42.4 % for spirotetramat -ketohydroxy and 29.3 – 43.8 % for spirotetramat-MA

-

### – 63 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

amide indicated that the SFO kinetic model was not the appropriate kinetic model. Further, the t-test probability of 0.25 for spirotetramat-MA-amide was not acceptable.

Therefore, bi-phasic models such as FOMC and DFOP w ere tested for spirotetramat-enol. The statistical results and the visual inspection of the fits demon strated that the FOMC fits represented a relatively appropriate kinetic model to describe the dissipati on of the test item (chi 2 

### scaled-error values in the range of 5.6 – 19.0 % for spirotetramat-enol (FOMC), 15.2 – 28.3 % for spirotetramat-ketohydroxy (SFO) and 29.1 – 45.3 % for spirotetramat-MA-amide (SFO)). The bi-ex ponential model DFOP was checked for the test item and was found to lead to an improved fit. The respe ctive chi 2 

### scaled-error values ranged from 2.4 – 5.6 % for spirotetramat-enol (DFOP), 19.2 – 36.8 % for spirot etramat-ketohydroxy (SFO) and 28.5 – 41.8 % for spirotetramat-MA-amide (SFO). Moreover the visual i nspection of the fits demonstrated that the DFOP fi ts represented the most appropriate kinetic model for the spirotetramat-enol dissipation in the four soil s.

Using the DFOP model, DT 50 

### values of spirotetramat-enol (best fit kinetic) in the four soils tested range from 0.02 – 0.18 days (non-normalized data). The DT 90 

### values in the four soils range from 10.9 – 40.9 da ys (summarized in Table B.8.1.2.3-2).

Table B.8.1.2.3-2: Best fit DT 50 

### and DT 90 

### values [days] of spirotetramat-enol based on DFOP k inetics (non-normalized data).

Soil DT 50 [days] DT 90

[days] DT 50recalc a

[days] χ 2 error [%] t-probability

‘Molino’ 0.06 28.9 8.7 5.5 < 0.001
‘Laacherhof AXXa’ 0.02 40.9 12.3 5.6 < 0.001
‘Laacherhof AIII’ 0.18 16.8 5.1 2.4 < 0.001
‘Hoefchen’ 0.02 10.9 3.3 5.2 < 0.001

a SFO-DT 50 recalculated from the DFOP-DT 90 divided by 3.32.



### For the estimation of modelling parameters the SFO- RB model was tested and was found to be the most suitable kinetic model which fitted the analyzed da taset best. Spirotetramat-enol is rapidly degraded in the equilibrium domain but also rapidly transferred int o a stronger bound soil compartment (‘protected compartment’), where it is not susceptible to degra dation and only slowly transferred back into the equilibrium domain from which it degrades to spirot etramat-ketohydroxy following SFO kinetics. The vis ual
assessments and the calculated scaled errors indica ted acceptable fits. The range of the chi 2 

### error is 6.4 – 11.0 % for spirotetramat-enol (SFO-RB), 17.7 – 33.8 % for spirotetramat-ketohydroxy (SFO) and 27.4 – 40.0 % for spirotetramat-MA-amide (SFO). From the s caled error and visual assessment the fits are excellent for spirotetramat-enol and altogether acc eptable for spirotetramat-ketohydroxy and spirotetr amatMA-amide. The t-test probability is < 0.001 for spi rotetramat-enol and spirotetramat-ketohydroxy and r anged from < 0.001 – 0.029 for spirotetramat-MA-amide, in dicating that all parameters are significant ( p < 0.05).

The DT 50/90 

### values were calculated from optimized first-order r ate constants and were normalized to the soil moisture at field capacity using estimated moisture correction factors (see Table B.8.1.2.3-1). The
normalized DT 50 

### values ranged from 0.02 – 0.13 days for spirotetra mat-enol, from 1.2 – 12.1 days for spirotetramat-ketohydroxy and from 0.2 – 3.9 days f or spirotetramat-MA-amide (summarized in Table B.8.1.2.3-7).

As the estimated biphasic SFO-RB half-lives of spir otetramat-enol are not applicabel to be used for e. g. 1 st



### tier PEC Soil 

### and PEC SW 

### model approaches (EU risk assessment) the recalcul ated DT 50 

### value from the DT 90 

### value of the biphasic FOMC model (SFO-DT 50 

### = FOMC-DT 90 

### / 3.32) is suggested for modelling purposes. The visual assessments of the FOMC fits were accept able and the calculated scaled chi 2 

### errors ranged from 5.6 – 19 % for spirotetramat-enol.

The kinetic evaluations for all four soils are summ arized in the tables B.8.1.2.3-3 to B.8.1.2.3.-6.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.2.3-3: Kinetic evaluation of the spirot etramat-enol degradation study based on SFO  SFO  SFO kinetics for spirotetramat-enol, spirotetramat -ketohydroxy and spirotetramat-MAamide, respectively.

Compound Kinetics Parameter ‘Molino’ ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’

DT 50 [days] 0.098 0.022 0.22 0.020

DT 50 norm a [days] 0.068 0.017 0.16 0.013

DT 90 [days] 0.33 0.072 0.73 0.066

χ 2 error [%] 28.8 37.4 20.9 31.3

R 2 0.98 0.98 0.98 0.99Spirotetramatenol SFO

t-probability < 0.001 < 0.001 < 0.001 < 0.001

DT 50 [days] 16.6 4.14 4.87 1.49
DT 50 norm a [days] 12.0 3.68 3.94 1.15

DT 90 [days] 55.1 13.8 16.2 4.93 Formation fraction [0 ... 1] 0.19 0.19 0.33 0.22

χ 2 error [%] 31.1 24.2 37.8 42.4

R 2 0.79 0.93 0.86 0.86Spirotetramatketohydroxy SFO

t-probability 0.010 0.022 < 0.001 0.017

DT 50 [days] 5.32 1.09 1.76 0.30
DT 50 norm a [days] 3.83 0.97 1.43 0.23

DT 90 [days] 17.7 3.62 5.86 0.99 Formation fraction [0 ... 1] Fixed to 1.0 during kine tic evaluation

χ 2 error [%] 33.8 29.3 37.5 43.8

R 2 0.76 0.86 0.89 0.82SpirotetramatMA-amide SFO

t-probability 0.18 0.22 0.04 0.25

a DT 50 normalized to 20 °C and 100 % FC (pF2).

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.2.3-4: Kinetic evaluation of the spirot etramat-enol degradation study based on FOMC  SFO  SFO kinetics for spirotetramat-enol, spirotetramat -ketohydroxy and spirotetramatMA-amide, respectively.

Compound Kinetics Parameter ‘Molino’ ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’

DT 50 [days] 0.060 0.018 0.12 0.016
DT 90 [days] 3.64 9.78 5.81 2.63

DT 50recalc a [days] 1.10 2.95 1.75 0.79

DT 50recalc norm ab [days] 0.80 2.64 1.42 0.61 alpha 0.4116 0.2594 0.4374 0.3235
beta 0.0136 0.0014 0.0302 0.0021

χ 2 error [%] 18.97 16.0 5.6 16.9

R 2 0.97 0.97 0.99 0.98Spirotetramatenol FOMC

t-probability nc nc nc nc

DT 50 [days] 11.7 3.5 4.6 1.3
DT 50 norm b [days] 8.5 3.1 3.7 1.0

DT 90 [days] 38.9 11.8 15.3 4.4 Formation fraction [0 ... 1] 0.22 0.24 0.35 0.27

χ 2 error [%] 22.5 15.2 16.8 28.3

R 2 0.90 0.98 0.96 0.92Spirotetramatketohydroxy SFO

t-probability nc nc nc nc

DT 50 [days] 3.2 0.9 1.7 0.3
DT 50 norm b [days] 2.3 0.8 1.4 0.2

DT 90 [days] 10.8 3.0 5.5 0.8 Formation fraction [0 ... 1] Fixed to 1.0

χ 2 error [%] 39.4 29.1 38.0 45.3

R 2 0.71 0.87 0.88 0.75SpirotetramatMA-amide SFO

t-probability nc nc nc nc

a SFO-DT 50 recalculated from FOMC-DT 90 divided by 3.32.
b DT 50 normalized to 20 °C and 100 % FC (pF2).

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.2.3-5: Kinetic evaluation of the spirot etramat-enol degradation study based on DFOP  SFO  SFO kinetics for spirotetramat-enol, spirotetramat -ketohydroxy and spirotetramatMA-amide, respectively.

Compound Kinetics Parameter ‘Molino’ ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’

DT 50 [days] 0.06 0.02 0.18 0.02
DT 90 [days] 28.9 40.9 16.8 10.9

DT 50recalc a [days] 8.7 12.3 5.1 3.3

DT 50recalc norm ab [days] 6.3 10.9 4.1 2.5
k 1 [days -1 ] 15.7 40.5 5.20 41.8
k 2 [days -1 ] 0.011 0.0098 0.030 0.011

g c [%] 0.86 0.85 0.83 0.89

χ 2 error [%] 5.54 5.6 2.4 5.2

R 2 0.99 0.99 0.99 0.99Spirotetramatenol DFOP

t-probability < 0.001 < 0.001 < 0.001 < 0.001

DT 50 [days] 16.7 4.2 5.1 1.5
DT 50 norm b [days] 12.1 3.8 4.1 1.2

DT 90 [days] 55.6 13.8 16.9 5.0 Formation fraction [0 ... 1] 0.21 0.22 0.36 0.24

χ 2 error [%] 24.2 19.2 28.8 36.8

R 2 0.86 0.96 0.9 0.89Spirotetramatketohydroxy SFO

t-probability < 0.001 < 0.001 < 0.001 < 0.001

DT 50 [days] 5.5 1.1 1.9 0.3
DT 50 norm b [days] 4.0 1.0 1.5 0.2

DT 90 [days] 18.1 3.6 6.3 1 Formation fraction [0 ... 1] Fixed to 1.0

χ 2 error [%] 30.8 28.5 30.9 41.8

R 2 0.77 0.87 0.91 0.82SpirotetramatMA-amide SFO

t-probability 0.005 < 0.001 0.001 0.029

a SFO-DT 50 recalculated from DFOP-DT 90 divided by 3.32.
b DT 50 normalized to 20 °C and 100 % FC (pF2).

c g denotes the size [0 ... 1] of the fast degrading fraction.

### – 67 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.2.3-6: Kinetic evaluation of the spirot etramat-enol degradation study based on SFO-RB  SFO  SFO kinetics for spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramatMA-amide, respectively.

Compound Kinetics Parameter ‘Molino’ ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’

C 0 100.6 100.3 96.2 100.7
k 1 2.786 7.701 1.548 9.032
k 2 2.145 6.045 0.859 4.701
k 3 0.013 0.012 0.037 0.012
k 4 0.041 0.166 0.136 0.456
k 5 0.127 0.632 0.365 2.222
k 6 10.762 26.780 2.798 28.982 Rates in the overall model -

k 1 + k 6 13.547 34.481 4.346 37.114

DT 50 a [days] 0.05 0.02 0.16 0.02
DT 50 norm b [days] 0.04 0.02 0.13 0.02

DT 90 [days] 0.17 0.07 0.53 0.06

χ 2 error [%] 6.75 6.8 10.98 6.36

R 2 0.997 0.997 0.993 0.996 t-probability < 0.001 < 0.001 < 0.001 < 0.001

K Lin eq c 0.790 0.540 0.780 0.700Spirotetramatenol SFO-RB

f ne d 193.2 693.1 28.9 593.7

DT 50 [days] 16.7 4.2 5.1 1.5

DT 50 norm b [days] 12.1 3.8 4.1 1.2

DT 90 [days] 55.6 13.9 16.9 5.1 Formation fraction [0 ... 1] 0.21 0.22 0.36 0.24

χ 2 error [%] 22.3 17.7 27.2 33.8

R 2 0.859 0.956 0.874 0.889Spirotetramatketohydroxy SFO

t-probability < 0.001 < 0.001 < 0.001 < 0.001

DT 50 [days] 5.4 1.1 1.8 0.3

DT 50 norm b [days] 3.9 1.0 1.5 0.2

DT 90 [days] 18.1 3.6 6.3 1 Formation fraction [0 ... 1] Fixed to 1.0 during kine tic evaluation

χ 2 error [%] 29.6 27.4 29.6 40

R 2 0.771 0.866 0.913 0.823SpirotetramatMA-amide SFO

t-probability 0.0047 < 0.001 0.001 0.029

a DT 50 resulting from sum of degradation rates k 1 and k 6 b DT 50 normalized to 20 °C and 100 % FC (pF2).

c K Lin eq = linearised sorption coefficient (FOCUS, 2006), equ ivalent to the K d value obtained in the column leaching study (Babcz inski, 2006c)
d f ne = factor for describing the ratio between the equi librium and non-equilibrium Freundlich coefficients (FOCUS, 2006)

### – 68 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion: The kinetic evaluation of the aerobic soil degradat ion of the metabolites spirotetramat-enol, spirotet ramatketohydroxy and spirotetramat-MA-amide was performe d according to the FOCUS (2006) recommendations. Spirotetramat-enol, main metabolite of spirotetrama t, spirotetramat-ketohydroxy and spirotetramat-MA-a mide degraded quickly and are expected not to accumulate in soil.

Table B.8.1.2.3-7: Summary on normalized DT 50 

### values of spirotetramat-enol, spirotetramatketohydroxy and spirotetramat-MA-amide according to the SFO-RB  SFO  SFO modelling approach (best fit model).

DT 50 normalized to 20°C and pF2

Spirotetramat-enol (SFO-RB) Spirotetramatketohydroxy (SFO) SpirotetramatMA-amide (SFO) Soil

DT 50 norm

[days] f ne [0 ... 1] k d a [days -1 ] DT 50 norm [days] DT 50 norm

[days]

‘Molino’ 0.04 193.2 0.013 12.1 3.9
‘Laacherhof AXXa’ 0.02 693.1 0.012 3.8 1.0
‘Laacherhof AIIII’ 0.13 28.9 0.037 4.1 1.5
‘Hoefchen’ 0.02 593.7 0.012 1.2 0.2

a k d (PEARL) = k 3 (SFO-RB)



### Reviewer comments:

• Outlier (spirotetramat-enol, in soil Laacherhof AI II, DAT-4) removed was not justified in the origina l report (Babczinski, 2006b), but is accepted by the reviewer.

• It was accepted by the reviewer that only the spir otetramat-enol study was considered relevant for the degradation kinetics calculations of the spirot etramat metabolites. In a qualitative consistency check (simulated degradation of spirotetramat and i ts metabolites using the derived kinetic parameter set plotted against the measured data for the spiro tetramat-parent study) it was shown that the simulated results appeared to be plausible in relat ion to the measured data also in the spirotetramat degradation study. However, comparatively high resi duals especially in case of spirotetramatketohydroxy and spirotetramat-MA-amide demonstrated that due to the analytical bias of the metabolite data these fits should not be used quant itatively.

• Beside the SFO-RB approach, the DFOP kinetics is t he ‘best fit’ model for the degradation of spirotetramat-enol. However, the notifier derived 1 st 

### tier SFO-DT 50 

### values for spirotetramat-enol from FOMC kinetics (recalculated from the FOMC-DT 90 

### ). Following the FOCUS kinetic report, recalculated
SFO-DT 50 

### should be based on FOMC kinetics, if i) FOMC kinet ics yields a sufficient fit to the measured data and ii) if less than 10 % of the pare nt (spirotetramat-enol) remains by study termination. Therefore, the approach suggested by t he notifier is accepted by the reviewer.

### – 69 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: [Carbonyl- 14 

### C]-4-Methoxycyclohexanone: Aerobic Soil Degradation in Three
Soils Author(s), year: Dehner, D., Heinemann, O., 2006 Study/report number: M-269022-02-2, MEF-05/485, ame nded 18.07.2006 Guideline(s): EPA Guideline for Aerobic Soil Metabolism, Subdivis ion N, Section 162-1,
EC-Directive 91/414/EEC Annex I Part 7 and Annex II Part 9,
OECD Guideline 307: Aerobic and Anaerobic Transform ation in Soil (2002)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: [Carbonyl- 14 

### C]-4-methoxy-cyclohexanone (CAS name: cyclohexanone , 4- methoxy); sample ID: BECH 1831 (reference synthesis : KML 3292-3), specific activity: 4.71 MBq mg -1 

### (= 127.31 μCi mg -1 

### ); radiochemical purity: 97.0 % (acc. radio-HPLC); Chemical purity: 96.9 % (GC-FID) Reference substances: [Carbonyl- 14 

### C]-4-methoxycyclohexanone, no other reference items were used Test system: Static incubation test system Acclimatisation time: 5 days
Test conditions: Aerobic conditions were maintained . Moisture adjusted soils (55 % MWHC) were placed in the dark at 20 °C. At DAT-0 the absolute microbial biomass carbon contents corresponded to 0.5 %, 2.6 % and 3.2 % rel ative to the soil organic carbon content of the soils ‘Molino’, ‘Laacherhof A III’ and ‘Hoefchen’, respectively. Number of replications: Duplicate samples
Test duration: Soil ‘Hoefchen’ and ‘Laacherhof AIII ’: 14 days Soil ‘Molino’: 3 days (because of the fast degradat ion observed in the EU soils) Sampling intervals: Soil ‘Hoefchen’ and ‘Laacherhof AIII’: 0 (approx. 1 h after treatment), 1, 3, 7 an d 14 days Soil ‘Molino’: 0 (approx. 1 h after treatment), 3 h , 7 h, 1, 2 and 3 days Application: 0.133 mg ai kg -1 

### dry soil (nominal) equivalent to 50 g ai ha -1 

### (nominal, calculation based on homogeneous distribution for 2.5 cm soil d epth and 1.5 g/cm 3 

### soil density), supplied test item was dissolved in water and drop wise applied to the soil surface. No instability in the stock solution was observed. Analyses: All soil samples were extracted directly at the day of sampling.
Extraction Procedure EU soils: Extraction was done by shaking three times at room temperature with 80 mL of acetonitrile/water/formic acid (100/100/0.5, v/v/v) , then twice with 80 mL of acetonitrile/1 M aqueous HCl (1/1, v/v) and once wi th 80 mL of pure acetonitrile, using a mechanical shaker. Due to unforeseen radioactivity losses the extracti on procedure was changed at the last two sampling days (DAT-7 and DAT-14): The first extraction was done by shaking twice with 80 mL of acetonitrile/1 M aqueou s HCl (1/1, v/v) in the test flask fitted with a trap for volatiles. After each shaking period the test flasks were placed into a desiccator to transfer 14 

### CO 2 

### into the trap (at day 7 a new trap for volatiles was placed on the test flask). Soils were further extracted by three cycles with 80 mL of acetonitrile/water/formic acid (100/1 00/0.5, v/v/v), one cycle with 80 mL of acetonitrile/1 M aqueous HCl (1/1, v/v) and o nce with 80 mL pure acetonitrile, using a mechanical shaker.
Extraction Procedure US soil:

### – 70 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

80 mL of acetonitrile/1 M aqueous HCl (1/1, v/v) we re poured into the incubation system fitted with a trap for volatiles



## . After each shaking period the test flasks were placed into a desiccator to transfer 14 

### CO 2 

### into the trap. Soils were further extracted by one cycle with 80 mL of acetonitrile/1 M aqueous HCl (1/1, v/v), three cycles with 80 mL of acetonitrile/water/formic acid (100/100/0.5, v/v/v) and one cycle with 80 mL pure acetonitrile, using a mechani cal shaker.

The combined extractable residues were radio-assaye d by LSC and analyzed by reversed-phase HPLC (HPLC-flow-through 14 

### C-radioactivity detection), with exemplary result verification in normal phase TLC ( TLC- 14 

### C-phosphor-imaging).

In order to determine the non-extractable soil resi dues air-dried extracted soils and paper filters were combusted and 14 

### C levels were measured using LSC. Volatile traps: Static volatile trap attachment for CO 2 

### (soda lime) and organic volatiles (polyurethane foam), permeable for oxygen (identifi cation of 14 

### C as 14 

### CO 2 

### by
BaCO 3 

### -precipation) Analytical techniques: LSC, RP-HPLC, Si-60 silica T LC, LC-MS
Detection limits: LSC: for raw extracts LOQ ≤ 0.55 % of AR, for combustion LOQ ≤ 0.05 % of AR;
for CO 2 

### samples LOQ < 0.01 % of AR; HPLC: LOD = 0.9 % of A R, LOQ was set
to three times the LOD: 2.7 % of AR. TLC: LOD = 1.5 % of AR, LOQ was set to
three times the LOD: 4.5 % of AR Kinetic evaluation: SFO and FOMC kinetics, MatLab v ers. 7.0.4.365; values < LOD were set to 0.5
LOD. Values between LOD and LOQ were set to the mea sured values.

Table B.8.1.2.3-8: Main soil properties.

Property ‘Hoefchen’ ‘Laacherhof AIIIa’ ‘Molino’

Location Germany/Burscheid Germany/Monheim USA/Pensacola Texture (USDA) Silt loam Loam Loamy sand

Soil taxonomic classification (USDA) Loamy, mixed, mesic Typic Argudalfs Loamy, mixed, mesic Typic Argudalfs Fine-loamy, kaolinitic, thermic Plinthic Kandiudults SCS soil series Alfisols, Udalfs Alfisols, Udalfs Ultis ol Sand (USDA) [%] 29.0 43.0 78.4 Silt (USDA) [%] 52.0 34.0 14.5 Clay (USDA) [%] 19.0 23.0 7.1 pH (H 2 O) 6.8 7.0 6.7 pH (CaCl 2 ) 6.5 6.6 5.9 pH (saturated paste) 6.9 7.0 6.4 Organic matter a [%] 4.1 1.7 1.2 Organic carbon [%] 2.4 1.0 0.7 CEC [meq/100 g DM] 12.8 9.3 5.0 Moisture content [g/100 g DM] MWHC WHC at 1/3 bar 65.0 20.6 45.1 13.0 34.6 8.8
Biomass b [mg microbial C/kg soil DM] start DAT-0: 756 DAT-0: 260 DAT-0: 32
Biomass b [mg microbial C/kg soil DM] end DAT-14: 670 DAT-59: 270 DAT-3: N/A

a Organic C = organic matter / 1.724
b Substrate induced respiration N/A not applicable

### – 71 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Findings: All values are given as mean values of the replicat es.

The material balances ranged from 71.3 % to 97.8 % of AR for ‘Hoefchen’ soil, from 70.4 % to 99.0 % fo r ‘Laacherhof AIII’ soil and from 95.4 % to 99.7 % of AR for ‘Molino’ soil, respectively. According to t he study authors the losses of radioactivity in soils ‘Hoefc hen’ and ‘Laacherhof’ at days 1 and 3 was caused by
liberation of 14 

### CO 2 

### bound in the soil during the extraction procedure using acidic solvent. Therefore the following extraction procedure was changed whereas the first extraction step was performed with test systems closed with a soda lime trap. It was shown that the following balances were found > 96 % of AR

.

In all three tested soils a high release of 14 

### CO 2 

### was observed reaching levels up to 74.1 %, 75.8 % and 66.3 % of AR (in soils ‘Hoefchen’, ‘Laacherhof AIII ’ and ‘Molino’, respectively) at study termination. No significant amounts (< 0.2 % of AR) of other volati le compounds were detected.

A rapid decrease of extractable residues occurred d uring the first incubation days in soils ‘Hoefchen’ and ‘Laacherhof AIII’ (‘Hoefchen’/’Laacherhof AIII’: fr om 97.1/98.6 % of AR at DAT-0 to 6.6/8.6 % of AR at DAT1). In soil ‘Molino’ the decrease was considerably slower than in the European soils.

All transformation products observed were minor whi ch amounts did not exceed 5 % of AR.

The portion of non-extractable radioactivity rapidl y increased reaching its peaks at levels of 22.6 to 24.9 % of AR after one to two days after application. A sligh t decrease in NER until study termination was obser ved.

The laboratory half-live of 4-methoxy-cyclohexanone in the European soils was < 1 day. It was not poss ible
to calculate statistical DT 50/90 

### values due to the fast dissipation and insufficient data points. For soil ‘Molino’ a
DT 50 

### of 0.6 days and a DT 90 

### of 1.8 days was calculated according to simple fir st order kinetics.

Table B.8.1.2.3-9: Distribution and recovery of app lied radioactivity [% of AR] after application of [carbonyl- 14 

### C]-4-methoxy-cyclohexanone to three soils (mean of duplicate samples).

Soil DAT Extractable CO 2 NER Total

0 97.1 na 0.7 97.8

1 6.6 39.7 24.9 71.3
3 4.5 55.1 22.6 82.3

7 3.9 70.1 22.3 96.3 ‘Hoefchen’

14 3.2 74.1 20.2 97.5

0 98.6 na 0.4 99.0

1 8.6 37.4 24.3 70.4
3 5.0 49.7 22.6 77.5

7 4.1 72.8 20.9 97.8 ‘Laacherhof AIIIa’

14 3.2 75.8 19.6 98.6

0 99.5 na 0.2 99.7
0.13 94.9 2.9 1.6 99.3
0.29 82.1 10.4 4.8 97.2

1 31.9 46.6 18.3 96.8
2 10.8 62.0 22.6 95.4 ‘Molino’

3 12.2 66.3 18.9 97.4 na denotes not analysed

### – 72 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.2.3-10: Characterization of extractable radioactivity [% of AR] after application of [carbonyl- 14 

### C]-4-methoxy-cyclohexanone to three soils (individu al replicates).4-Methoxycyclohexanone ROI 1 ROI 2 ROI 3 ROI 4 Soil DAT
A B A B A B A B A B

0 92.1 96.3 3.0 2.8 nd nd nd nd nd nd

1 nd 0.9 nd nd 2.0 2.0 1.0 1.0 2.8 2.7
3 nd nd nd nd nd nd 1.4 1.4 3.2 3.1

7 nd nd nd nd nd nd 1.8 0.9 2.1 2.9 ‘Hoefchen’

14 nd nd nd nd nd nd 1.4 nd 1.6 2.6

0 99.6 97.7 nd nd nd nd nd nd nd nd

1 nd 1.0 nd nd 3.3 2.6 1.4 1.2 3.2 4.0
3 nd nd nd nd nd nd 1.5 1.4 3.5 3.7

7 nd nd nd nd nd nd 2.1 1.0 2.0 3.1 ‘Laacherhof AIIIa’

14 nd nd nd nd nd nd nd 1.6 2.5 1.6

0 98.4 100.6 nd nd nd nd nd nd nd nd
0.13 94.3 95.5 nd nd nd nd nd nd nd nd
0.29 81.5 82.7 nd nd nd nd nd nd nd nd

1 23.1 22.3 nd nd 4.4 4.7 nd nd 3.6 2.9
2 3.9 3.6 nd nd 2.7 2.6 nd nd 3.7 3.5 ‘Molino’

3 8.8 2.4 nd nd 2.4 2.5 nd nd 4.3 2.4

nd denotes not determined



### Table B.8.1.2.3-11: Calculated DT 50 

### and DT 90 

### of [carbonyl- 14 

### C]-4-methoxy-cyclohexanone in three soils using SFO and FOMC kinetics.

SFO Soil DT 50 [days] DT 90 [days] Best fit model, χ 2 error
‘Hoefchen’ < 1 < 1 not evaluated a

‘Laacherhof’ < 1 < 1 not evaluated a

‘Molino’ 0.6 1.8 SFO, 11.2 %

a For the European soils no statistical DT 50 /DT 90 values were calculated due to the very short dissi pation and therefore the lack of data points after the treatment.



### Conclusion: The photolysis metabolite 4-methoxy-cyclohexanone d egrades very quickly in all three tested soils unde r aerobic conditions. The half-live is < 1 day. No ma jor degradates ( ≥ 5 % of AR) were detected. The major transformation product was 14 

### CO 2 

### formed with maximum amounts of 66.3 % to 75.8 % of AR at study
termination. The NER fraction increased fast to its peak level up to ~25 % of AR in soils ‘Hoefchen’ a nd
‘Laacherhof AIII’ and 23 % of AR in soil ‘Molino’. At study termination the NER declined to ≤ 20 % of AR. No volatile organic compounds were formed during the i ncubation time.

Reviewer comments:

• Microbial biomass carbon content in ‘Molino’ soil is rather low (corresponding to 0.5 % of total carb on content). According to the OECD guideline 307 the m icrobial biomass of at least 1 % of total organic
carbon is recommended.

### – 73 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.2.4 Anaerobic degradation rate of the active substance (OECD Annex IIA, 7.2.4, IIIA, 9.1.2)

The anaerobic degradation rate of spirotetramat was investigated in one study which was also used to elucidate the route of degradation under aerobic co nditions:

• Aerobic/ anaerobic degradation in one soil (20 °C, [3- 14 

### C] label)

Full details including degradation rates are given under B.8.1.1.2.

B.8.1.2.5 Anaerobic degradation rate of relevant me tabolites (OECD Annex IIA, 7.2.5)

The anaerobic degradation rates of relevant metabol ites of spirotetramat were investigated in one stud y which was also used to elucidate the route of degra dation under aerobic conditions:

• Aerobic/ anaerobic degradation in one soil (20 °C, [3- 14 

### C] label)

Full details are given under B.8.1.1.2.

### – 74 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.2.6 Summary on the rate of degradation in lab oratory soil studies

Note: In contrast to the EU risk assessment, the Canadia n and US risk assessment is based on nonnormalized soil degradation rates, and deviating in put data are used for calculation and modelling of predicted/expected environmental concentrations (e. g. the 80 th 

### percentile, or the 90 th 

### percentile of the confidence interval (C.I.) on the arithmetic mean, etc.). Respective values are stated in the summary tables together with the endpoints considered relevant for the EU risk assessment.

The laboratory soil degradation rate of spirotetram at and metabolites was investigated in 3 EU soils a nd 1 US soil with the following range of soil properties (pH, organic C, clay content) using [3- 14 

### C] labelled spirotetramat or [5- 14 

### C] and [3- 14 

### C] labelled spirotetramat-enol as parent.

• pH (CaCl 2 

### ) 5.4 – 6.7

• organic C 0.93 – 2.1 %

• clay content 5.0 – 12.0 %

No degradation rates are available for more alkalin e soil (with a pH > 7). However, degradation of spirotetramat was extremely fast in the pH range te sted and no dependency on soil pH was observed. Fro m hydrolysis behaviour it can be deduced that degrada tion of spirotetramat is likely to be even faster a t higher pH values. In field studies conducted on 4 US soils with a pH range of 7.0 – 8.1 (pH measured in water

, equivalent to approx. pH 5.5 – 7.3 in CaCl 2 

### ), no dependence on soil pH was observed. In this r espect, it is highly unlikely that degradation of spirotetramat u nder more alkaline conditions will significantly di ffer from degradation rates observed under more acidic condit ions.

Under aerobic conditions , spirotetramat was found to be a very fast degradi ng compound following biphasic degradation. Following best fit kinetics ( i.e. double first order in parallel kinetics, DFOP) , nonnormalized degradation half-life (DegT 50 

### ) values of spirotetramat were in a range of 0.09 – 0.30 days (chi 2



### error ≤ 6.1 %); respective DegT 90 

### was in a range of 0.34 – 1.26 days. Since most env ironmental models are not capable to handle DFOP kinetics, a conservative SFO-DegT 50 

### for modelling may be derived from the DFOP-DegT 90 

### divided by 3.32. This procedure results in a DegT 50recalc 

### in the range of 0.10 – 0.38 days.

Applying SFO kinetics (chi 2 

### error ≤ 21.8 % for one soil, for three soils chi 2 

### error ≤ 9.5 %), non-normalized DegT 50 

### values of spirotetramat were in a range of 0.08 – 0.33 days, respective DegT 90 

### values were in a range of 0.28 – 1.09 days. Following normalization to 20 °C and pF2, obtained DegT 50norm 

### values were in a range of 0.05 – 0.23 days.

The biodegradation of spirotetramat was additionall y investigated in two soils under outdoor climatic
conditions considered more realistic for the indent ed use (metabolism studies). In close agreement to the lab results, the parent compound was quickly degraded w ith a DT 50 

### of 1.2 and 2.9 days (‘Laacherhof AIIIa’ and ‘Molino’ soil, respectively, both SFO kinetics, R 2 

### ≥ 0.98).

Owing to analytical reasons (instability of spirote tramat-enol during acidic extraction), degradation rates of the metabolites spirotetramat-enol, spirotetramat-k etohydroxy and spirotetramat-MA-amide were obtained from the additional spirotetramat-enol laboratory d egradation study, only. This study was conducted on the same soils that were used for the parent degradatio n study. However, a slightly alkaline extraction pr ocedure was applied to ensure spirotetramat-enol stability during extraction.

Similar to the parent, spirotetramat-enol dissipated following pronounced biphasic kinetics, with an extremely quick first phase and a significantly slo wer second phase. This extremely biphasic behaviour of spirotetramat-enol can be explained by an extensive oxidation of the compound leading either to format ion of

### – 75 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

spirotetramat-ketohydroxy or to extensive binding t o the soil matrix. The ‘nature’ of the extensive bi nding to the soil matrix is not fully understood, yet. Howev er, binding of spirotetramat-enol to the soil matri x is partly reversible under conditions of extraction, releasin g a fraction which mainly contributes to the second and slower phase of the biphasic degradation kinetics. Both processes (formation of spirotetramat-ketohydr oxy and binding to the soil matrix) are considered to t ake place almost immediately once in contact with s oil (under aerated conditions, oxidation process) and a re likely to be competitive factors depending on th e available oxygen.

The best fit kinetic model for degradation of spiro tetramat-enol is the DFOP model (chi 2 

### error ≤ 5.6 %). This model yielded a DegT 50 

### for spirotetramat-enol in the range of 0.02 – 0.18 days. Respective DegT 90 

### values were in a range of 10.9 – 40.9 days. Since most env ironmental models are not capable to handle DFOP kinetics, a conservative SFO-DegT 50 

### for modelling may be derived from the DFOP-DegT 90 

### divided by 3.32. This procedure results in a DT 50recalc 

### in the range of 3.3 – 12.3 days.

According to first order multi compartment (FOMC) k inetics, DegT 50 

### values of spirotetramat-enol were in a range of 0.02 – 0.12 days (chi 2 

### error ≤ 19.0 %), respective DegT 90 

### values were in a range of 2.6 – 9.8 days. Similar to the DFOP kinetics, a conservative SFO-De gT 50 

### for modelling may be derived from the FOMCDegT 90 

### divided by 3.32. This procedure results in a DegT 50recalc 

### in the range of 0.8 – 3.3 days. Following normalization to 20 °C and pF2, SFO-DegT 50recalc 

### values were in a range of 0.6 – 2.6 days. Since re sidues of spirotetramat-enol were clearly < 10 % of AR by stu dy termination, the normalized SFO-DegT 50recalc 

### deriving from FOMC kinetics is recommended by FOCUS as suita ble and conservative parameter for 1 st 

### tier modelling if a higher tier approach (e.g. SFO-RB ki netics, see following test) is not applicable (EU r isk assessment only).

The highly specific degradation behaviour of spirot etramat-enol was considered to be adequately explai ned by a simple first order-reversible binding (SFO-RB) model, which is equivalent to the (non-equilibrium ) kinetic sorption model in PEARL. In the SFO-RB model, (extr actable) spirotetramat-enol is considered to consis t of two fractions, one is mobile and degradable (to spi rotetramat-ketohydroxy), the other fraction is tigh tly bound to the soil matrix (where it is protected against f urther degradation assuming zero degradation). Sinc e binding to the soil matrix is considered reversible , a slow and steady re-transfer of bound spirotetra mat-enol into the mobile and degradable spirotetramat-enol f raction is assumed. Degradation of the mobile fract ion of spirotetramat-enol and transfer rates between mobil e and bound fraction are considered to follow SFO kinetics. The re-transfer (desorption) rate of the immobile spirotetramat-enol into the mobile fractio n is adequately reflected by the PEARL parameter k d 

### (kinetic sorption rate constant, [days -1 

### ]), the ratio between
immobilization and mobilization rates is mirrored b y the PEARL parameter f ne 

### (ratio between non-equilibrium and equilibrium sorption sites, dimensionless).

Consequently, residue data from the spirotetramat-e nol degradation study were fitted to a multi-compar tment model with spirotetramat-enol (parent) following SF O-RB kinetics and spirotetramat-ketohydroxy and spirotetramat-MA-amide linked in series (both follo wing SFO kinetics).

According to the SFO-RB approach, DegT 50 

### of spirotetramat-enol (degradation of the mobile and degradable fraction) was in a range of 0.02 – 0.16 days (SFO kinetics, chi 2 

### error ≤ 11.0 %, R 2 

### ≥ 0.99). The DegT 50 

### of the transfer rate into the stronger bound fract ion was estimated to be in a range of 0.11 – 0.18 days, the DegT 50 

### of the slow re-transfer back from the bound into t he degradable fraction was in a range of 18.7 – 57.8 days (the latter transfer rate is equiv alent to the desorption rate, K d 

### , required for kinetic adsorption modelling in PEARL). From the ratio of t he two transfer rates, the f ne 

### value can be deduced which is needed as further input value for kinetic adsorp tion modelling in PEARL. For spirotetramat-enol f ne 

### values were in a range of 29 – 693. Since formation fracti on of spirotetramat-enol from the parent, i.e. spir otetramat, is not known, a worst case formation fraction of 10 0 % is assumed for environmental modelling purposes

.

### – 76 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Additional modelling of a soil column study conduct ed on four soils (Babczinski, 2006c, Hammel, 2007), based on a kinetic adsorption approach using the tr ansport model HYDRUS (conceptual equivalent to PEARL) clearly demonstrated that the SFO-RB/kinetic adsorption approach adequately describes the behaviour of spirotetramat-enol observed in the col umn leaching experiments.

Following the overall best-fit modelling approach ( i.e. SFO-RB  SFO  SFO kinetics for spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amid e, respectively), degradation DegT 50 

### of spirotetramatketohydroxy in soil was calculated to be in a range of 1.5 – 1 6.7 days (SFO kinetics, chi 2 

### error ≤ 33.8 %, R 2



### ≥ 0.86). Respective DegT 90 

### values were in a range of 5.1 – 55.6 days. Formati on fraction of spirotetramatketohydroxy from its precursor, spirotetramat-enol, was calculated to be in a range of 20.6 – 35.6 % w ith an
arithmetic mean of 25.7 %. Normalization to 20 °C a nd pF2 resulted in DegT 50 

### values in a range of 1.2 – 12.1 days.

According to the SFO-RB  SFO  SFO kinetics approach, degradation DegT 50 

### of spirotetramat-MAamide in soil was calculated to be in a range of 0.3 – 5 .4 days (SFO kinetics, chi 2 

### error ≤ 40.0 %, R 2 

### ≥ 0.77). Respective DegT 90 

### was in a range of 1.0 – 18.1 days. Formation fract ion of spirotetramat-MA-amide from its precursor, spirotetramat-ketohydroxy, was fixed to 100 % during fitting procedure. Normalization to 20 °C and pF2 resulted in DegT 50 

### values in a range of 0.2 – 3.9 days.

It may be annotated, that degradation rates and for mation fractions of spirotetramat-ketohydroxy and spirotetramat-MA-amide obtained from the more simpl istic SFO  SFO  SFO, FOMC  SFO  SFO or
DFOP  SFO  SFO kinetics approaches (lower tier approaches) ar e almost identical to the data obtained from the higher tier approach SFO-RB  SFO  SFO. This is reasonable since the release of spiro tetramatketohydroxy from spirotetramat-enol is almost exclu sively attributed to the initial and fast degradati on phase of the (mobile) spirotetramat-enol fraction. The fo rmation of spirotetramat-ketohydroxy from the secon d and slower phase of the strongly bound (immobile) spiro tetramat-enol fraction is considerably almost negli gible. In this respect, a reasonable groundwater risk asse ssment may be solely based on obtained SFO degradation rates for all, the parent and all metab olites including spirotetramat-enol. The strongly b ound spirotetramat-enol fraction in the soil is consider ed immobile (no potential for leaching) and does no t significantly attribute to the further formation of metabolites owing to its rather slow degradation.

In a separate degradation study, the soil photolysi s metabolite 4-methoxy-cyclohexanone was shown to degrade very quickly in all three tested soils unde r aerobic conditions. The half-live was < 1 day.– 77 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.2.6-1: Summary on laboratory non-normalized, recalculatedand normalized recalculatedDegT50and DegT90values [days] of spirotetramatand spirotetramat-enol (both used as parent in therespective studies) in soil according to DFOP (bestfit for both compounds), FOMC and SFOkinetics.DFOP (best fit kinetics) FOMC SFOCompound SoilDegT50[days]DegT90[days]DegT50recalca[days]Norm.DegT50recalc[days]DegT50[days]DegT90[days]DegT50recalca[days]Norm.DegT50recalc[days]DegT50[days]DegT90[days]Norm.DegT50[days]‘Laacherhof AXXa’ 0.24 0.89 0.27 0.21 0.23 1.04 0.31 0.24 0.21 0.69 0.17‘Laacherhof AIII’ 0.26 0.97 0.29 0.21 0.25 1.09 0.33 0.23 0.23 0.77 0.16‘Hoefchen’ 0.09 0.34 0.10 0.07 0.01 0.44 0.13 0.090.08 0.28 0.05‘Molino’ 0.30 1.26 0.38 0.26 0.26 2.61 0.79 0.55 0.33 1.09 0.23Geometric mean (EU)ncnc0.230.17ncnc0.320.230.190.630.1380th%ile (PMRA)ncnc0.33ncncnc0.51nc0.270.90ncSpirotetramat(parent in study)90th%ile C.I. on meanb(US-EPA)ncnc0.36ncncnc0.62nc0.300.98nc‘Laacherhof AXXa’ 0.02 28.9 12.3 10.9 0.02 9.8 2.952.64 0.022 0.072 0.017‘Laacherhof AIII’ 0.18 40.9 5.1 4.1 0.12 5.8 1.75 1.42 0.22 0.73 0.16‘Hoefchen’ 0.02 16.8 3.3 2.5 0.02 2.6 0.79 0.61 0.020 0.066 0.013‘Molino’ 0.06 10.9 8.7 6.3 0.06 3.6 1.10 0.80 0.0980.33 0.068Geometric mean (EU)ncnc6.55.2ncnc1.461.160.0550.180.03980th%ile (PMRA)ncnc10.1ncnc7.42.23nc0.150.49ncSpirotetramatenol(parent in study)90th%ile C.I. on meanb(US-EPA)ncnc10.6ncnc8.12.43nc0.170.56ncaConservative SFO-DT50recalculated from DFOP-DT90and FOMC-DT90, respectively, divided by 3.32.bCalculated by [arithmetic mean + (t90,n-1× SD) / n0.5], wherebyt90,n-1= one-sided studentstvalue atα= 0.1, SD = standard deviation and n = number of data available

### – 78 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.2.6-2: Summary on non-normalized and normalized DegT 50 

### and DegT 90 

### values [days] of spirotetramat-enol, spirotetramat-ketohydroxy and s pirotetramat-MA-amide in soil obtained from the degradation study using spirotetramat-enol as paren t (multi-compartment modelling accounting for SFO-RB kinetics of spirotetramat-enol).

Compound Soil DegT 50 [days] DegT 90

[days] Norm.

DegT 50 [days] f ne [-] k d

[days -1 ] Formation

fraction [0 ... 1] Kinetics

‘Laacherhof AXXa’ 0.02 0.07 0.02 693 0.012 - SFO-RB
‘Laacherhof AIII’ 0.16 0.53 0.13 28.9 0.037 - SFO-RB
‘Hoefchen’ 0.02 0.06 0.02 594 0.012 - SFO-RB
‘Molino’ 0.05 0.17 0.04 193 0.013 - SFO-RB Geometric mean (EU) 0.04 0.14 0.03 377 a 0.016 - 80 th %ile (PMRA) 0.09 0.31 nc - - -Spirotetramatenol (parent in study)

90 th %ile of C.I. on

mean b (US-EPA) 0.12 0.39 nc - - -

‘Laacherhof AXXa’ 4.2 13.9 3.8 - - 0.223 SFO
‘Laacherhof AIII’ 5.1 16.9 4.1 - - 0.356 SFO
‘Hoefchen’ 1.5 5.1 1.2 - - 0.243 SFO
‘Molino’ 16.7 55.6 12.1 - - 0.206 SFO Geometric mean (EU) 4.8 16.1 3.9 - - 0.257
80 th %ile (PMRA) 9.7 32.3 nc - - -Spirotetramatketohydroxy

90 th %ile of C.I. on

mean b (US-EPA) 12.4 41.2 nc - - -

‘Laacherhof AXXa’ 1.1 3.6 1.0 - - 1.00 c SFO
‘Laacherhof AIII’ 1.8 6.3 1.5 - - 1.00 c SFO
‘Hoefchen’ 0.3 1.0 0.2 - - 1.00 c SFO
‘Molino’ 5.4 18.1 3.9 - - 1.00 c SFO Geometric mean (EU) 1.3 4.5 1.0 - - 1.00
80 th %ile (PMRA) 3.2 11.0 nc - - -SpirotetramatMA-amide

90 th %ile of C.I. on

mean b (US-EPA) 4.0 13.4 nc - - -

a Arithmetic mean
b Calculated by [arithmetic mean + ( t 90,n-1 × SD) / n 0.5 ], whereby t 90,n-1 = one-sided students t value at α = 0.1, SD = standard deviation and n = number of data available

c Fixed to maximum (i.e. 1.00) during fitting proced ure

### – 79 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.3 Field studies (OECD Annex IIA, 7.3, IIIA, 9 .2)

B.8.1.3.1 Soil dissipation testing (OECD Annex IIA, 7.3.1, IIIA, 9.2.1)

Field dissipation of spirotetramat and metabolites was investigated in four US field trials:

• New York, US

• Florida, US

• California, US

• Washington, US

Reference: Terrestrial field dissipation of BYI08330 in New Yo rk Soil, 2004 Author(s), year: Lenz, M., 2006a Study/Report number: M-277191-01-1, MEFNY004 Guideline(s): US-EPA No. 164-1: Terrestrial field d issipation, PMRA Data Code No. 8.3.2.1
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: Spirotetramat (unlabelled) as oil d ispersion formulation BYI08330 100 OD containing 102.02 g ai L -1 

### , batch 08030/0110(0073)
Internal standard: 13 

### C-spirotetramat, 13 

### C-spirotetramat-enol, 13 

### C-spirotetramat-ketohydroxy and 13 

### Cspirotetramat-MA-amide Plot preparation and
maintenance: Bare ground plot (77.7 × 4.6 m, 360 m 2 

### ) divided into three subplots (25.9 × 4.6 m,
120 m 2 

### ), one control plot (7.3 × 4.6 m, 34 m 2 

### ), plots maintained fallow with nonselective herbicides (glyphosate, paraquat) Application: 439 g ai ha -1 

### as a liquid spray, 220 L ha -1 

### , liquid broadcast spray (boom sprayer) Application date: 14 th 

### June, 2004 Study duration: 542 days Irrigation: 14 irrigation events, seasonally applie d as needed, 13 – 14 mm each time Application
verification: • Whatman filter pads (24 cm diameter, 452 cm 2 

### ), two per subplot, immediately sampled after application

• soil pans (approx. 24 x 33 cm, 797 cm 2 

### ) with 450 g of soil, one per subplot, immediately sampled after application Sampling: 5 cores (5.7 cm diameter) from each subpl ot per sampling data, composited into one sample per soil depth, 122 cm sampling depth, 15 cm soil core length, soil cores were analysed until two residue free layers were fo und Sample processing: • filter pads: extracted with 5 ml formic acid and 4 95 ml of 4 g L -1 

### cysteine hydrochloride in 2.5 % ethyl acetate in water/aceto nitrile (50/50, v/v) solution, aliquot fortified with internal 13 

### C standard solution, filtered and measured by
LC/MS/MS

• Soil pans: same method used for the soil samples

• Soil samples: microwave extraction (twice) of 20 g soil with 50 ml of acetonitrile/water (50/50, v/v) containing 8 g L -1 

### cysteine hydrochloride and 5 % ethyl acetate at 70 °C for 30 min, aliquot fortif ied with internal 13 

### C standard solution, filtered and measured by LC/MS/MS (analyt ical method FN-002- S05-03) Sample storage length: Soil samples stored frozen for maximum 786 days (an alysis period: 20 th 

### October
2005 – 9 th 

### June 2006)

### – 80 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Monitoring: Daily weather conditions, soil moisture at 5 and 30 cm soil depth Analytical techniques: LC/MS/MS LOQ = 5 μg kg -1 

### (each analyte) LOD = 0.5 μg kg -1 

### (spirotetramat, spirotetramat-enol), 0.9 μg kg -1 

### (spirotetramatketohydroxy), 2.1 μg kg -1 

### (spirotetramat-MA-amide) Fortification solution: Mixture of spirotetramat, s pirotetramat-enol, spirotetramatketohydroxy and spirotetramat-MA-amide (fortificati on at 5.0 μg kg -1 

### each)
Kinetic evaluation: • Parent and total combined residues: simple first o rder (SFO) kinetics:

• Spirotetramat-enol and spirotetramat-ketohydroxy: Spirotetramat SFO 

###  (spirotetramat-enol + spirotetramat-ketohydroxy) SFO 

###  sink (no additional pathways to sink) (additional e valuation by the reviewer):

All data without normalization, MATLAB 7.0.4

Table B.8.1.3.1-1: Study site.
Location New York, USA Slope gradient: 0 % Depth to groundwater: > 1.8 m 30 years average rainfall: 974 mm 30 years mean temperature: 9.1 °C Weather conditions during study: Within 30 years no rmal level, total water input 122 % of average rain fall Study mean temperature: 8.1 °C (June 2004 – May 2005 ), 13.9 °C (June 2005 – December 2005) Study rainfall: 941 mm (June 2004 – May 2005), 728 m m (June 2005 – December 2005) Study irrigation: 80 mm (June 2004 – May 2005), 106 mm (June 2005 – December 2005) Total precipitation (rainfall + irrigation): 1021 m m (June 2004 – May 2005), 834 mm (June 2005 – Decem ber 2005) Crops grown: 2001: alfalfa, 2002: alfalfa, 2003: al falfa

Pesticides used: 2001: Fusilade, Balan, Buctril, Pursuit, Baythroid, Po st Plus, Kerb, Sencor, 2002: Baythroid, Buctril, Post Plus, 2003: Velpar, Bayt hroid, Kerb
Fertilizer: Not available Plot preparation: Tillage 

### Parent Spirotetramat-enol +

Spirotetramat-ketohydroxy Sink Parent Sink

Combined residues Sink

### – 81 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.3.1-2: Soil properties. Soil depth [cm] Property 0 – 15 15 – 30 30 – 45 45 – 60 60 – 75 75 – 90 90 – 105 105 – 122

Texture (USDA) Loamy sand Loamy

sand Loamy

sand Sand Sand Sand Sand Sand

Sand (USDA) [%] 85 85 85 89 89 91 91 91 Silt (USDA) [%] 6 7 9 6 6 5 7 7 Clay (USDA) [%] 9 8 6 5 5 4 2 2 pH (soil/water, 1/1, m/v) 6.3 6.3 5.8 5.3 5.1 5.2 5 .4 5.5 pH (0.01 M CaCl 2 ) 5.5 5.5 5.0 4.6 4.5 4.5 4.6 4.6 Organic carbon [%] a 0.75 0.70 0.29 0.23 0.17 0.06 0.06 0.12 CEC [meq kg -1 ] 45 45 43 38 34 29 27 30 Bulk density [g cm -3 ] 1.40 1.38 1.41 1.43 1.44 1.47 1.49 1.46 MC (1/3 bar) [%] 8.9 8.2 6.3 5.6 6.0 4.3 4.1 4.4

a Organic carbon = organic matter / 1.724



### Findings: Daily soil moisture monitored at a soil depth of 30 cm was above field capacity (at 1/3 bar) throughou t the study duration. Soil moisture at 5 cm was pushed be yond field capacity by frequent rainfall and irriga tion. Therefore, sufficient water was applied to the test plot to promote leaching of the test substance.

Mean recovery of fortified control soil samples (5. 0 μg kg -1 

### of each analyte) were 107, 112, 96 and 112 % for spirotetramat, spirotetramat-enol, spirotetramat-ke tohydroxy and spirotetramat-MA-amide, respectively.

Application verification filter paper and soil pans showed an average parent equivalent recovery of 94 % and 96 % of the application rate, respectively. Average 0 DAT concentration of spirotetramat was 74.1 μg k g -1



### . Spirotetramat and its transformation products were not found below 15 cm of soil depth. Spirotetramat degraded to < LOD (0.5 μg kg -1 

### ) by 14 DAT, major degradation products formed were spirotetramat-enol (maximum 46.0 μg ai kg -1 

### by 1 DAT declining to 4.2 μg ai kg -1 

### by study termination) and spirotetramatketohydroxy (26.0 μg kg -1 

### by 1 DAT declining to < LOD by study termination).– 82 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.3.1-3: Total average residues [μg kg-1] in soil samples (0 – 45 cm layer) collected fromNow York bare ground plot.Spirotetramat Spirotetramat-enol Spirotetramat-ketohydroxy Spirotetramat-MA-amideDAT0 – 15cm15 – 30cm30 – 45cmaTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmaTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmaTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmaTotal(0 – 45cm)-4< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD074.1 na na 74.1 8.7 na na 8.7 12.2 na na 12.2 < LODna na < LOD117.7 < LOD < LOD 17.746.0< LOD < LOD46.0 26.0< LOD < LOD26.0< LOD < LOD < LOD < LOD32.2a< LOD < LOD 2.2a40.3 < LOD < LOD 40.3 24.5 < LOD < LOD 24.52.7a< LOD < LOD2.7a71.3a< LOD < LOD 1.3a32.2 < LOD < LOD 32.2 22.6 < LOD < LOD 22.6 < LOD< LOD < LOD < LOD14< LOD < LOD < LOD < LOD 39.5 < LOD < LOD 39.5 16.9< LOD < LOD 16.9 < LOD < LOD < LOD < LOD28< LOD < LOD < LOD < LOD 16.8 < LOD < LOD 16.8 12.0< LOD < LOD 12.0 < LOD < LOD < LOD < LOD60< LOD < LOD < LOD < LOD 14.3 < LOD < LOD 14.3 3.0a< LOD < LOD 3.0a< LOD < LOD < LOD < LOD121< LOD < LOD < LOD < LOD 8.3 < LOD < LOD 8.3 1.5a< LOD < LOD 1.5a< LOD < LOD < LOD < LOD179< LOD < LOD < LOD < LOD 8.8 < LOD < LOD 8.8 1.3a< LOD < LOD 1.3a< LOD < LOD < LOD < LOD289< LOD < LOD < LOD < LOD 11.6 < LOD < LOD 11.6 1.8a< LOD < LOD 1.8a< LOD < LOD < LOD < LOD365< LOD < LOD < LOD < LOD 4.8a< LOD < LOD 4.8a1.2a< LOD < LOD 1.2a< LOD < LOD < LOD < LOD542< LOD < LOD < LOD < LOD 4.2a< LOD < LOD 4.2a< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LODaResidues < LOQ (5 μg kg-1)LOD = limit of detection (0.5 μg kg-1for spirotetramat and spirotetramat-enol, 0.9 μg kg-1for spirotetramat-ketohydroxy, 2.1 μg kg-1for spirotetramat-MA-amide)na = not analysed

### – 83 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.3.1-4: Average residues [μg kg -1 

### ] of spirotetramat and average parent equivalents [ μg ai equiv kg -1 

### ] of spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amide in soil samples from 0 – 45 cm colleted from New York bare ground plots. Bare ground plot DAT Spirotetramat Spirotetramat-enol Spirotetramatketohydroxy Spirotetramat-MAamide Total

-4 < LOD < LOD < LOD < LOD < LOD
0 74.1 10.8 14.4 < LOD 99.3

1 17.7 57.0 30.6 < LOD 105.3
3 2.2 49.9 28.8 2.7 83.6

7 1.3 39.9 26.6 < LOD 67.8

14 < LOD 48.9 19.9 < LOD 68.8
28 < LOD 20.8 14.1 < LOD 34.9
60 < LOD 17.7 3.5 < LOD 21.2

121 < LOD 10.3 1.8 < LOD 12.1
179 < LOD 10.9 1.5 < LOD 12.4
289 < LOD 14.4 2.1 < LOD 16.5
365 < LOD 5.9 1.4 < LOD 7.3
542 < LOD 5.2 < LOD < LOD 5.2



### Table B.8.1.3.1-5: Calculated dissipation half-life [days] of spirotetramat, spirotetramat-enol, spirotetramat-ketohydroxy and total combined residu es in New York bare ground soil (nonnormalized) based on SFO kinetics.

Study site Substance DT 50 [days] DT 90

[days] R 2 χ 2

error [%] Formation

fraction [0 ... 1] Kinetics

Spirotetramat 0.5 1.6 0.94 5.1 - SFO Sum of spirotetramat-enol + spirotetramat ketohydroxy b 31.6 105 nc 16.6 1.0 a All SFO in series New York bare ground

Total combined residues c 23.4 77.8 0.93 16.4 - SFO

a No additional pathways to sink assumed, therefore f ixed to 1.0 during modelling
b Calculated by the reviewer (see comments)

c Total combined residues: sum of spirotetramat, spi rotetramat-enol, spirotetramat-ketohydroxy, spirote tramat-MA-amide



### Conclusion: Spirotetramat dissipated rapidly under field condit ions on bare ground plots with a DT 50 

### of 0.49 days and a
DT 90 

### of 1.62 days (R 2 

### = 0.94, SFO kinetics, χ 2 

### error = 5.1 %). Residues of spirotetramat and its transformation products were not found > LOD below the 0 – 15 cm soil layer. Total combined residues dissipated with a DT 50 

### of 23.4 days. Combined residues of spirotetramat-e nol and spirotetramat-ketohydroxy degraded with a DT 50 

### of 31.6 days.

Reviewer comments:

• Owing to the distinct storage instability of spiro tetramat-enol (degrading to spirotetramat-ketohydro xy and bound to the soil matrix during freeze storage, see Netzband, 2006), residues of spirotetramatenol and spirotetramat-ketohydroxy should not be ev aluated individually.

• Samples were screened for spirotetramat-enol dimer 1 and spirotetramat-enol dimer 2 (position paper Hellpointner et al., 2007). Both dimers stay ed below a concentration of 0.2 % of applied during the whole course of the trial. In this respe ct, dimerization of spirotetramat-enol, frequently observed in laboratory studies after hot-spot appli cation, has to be considered negligible under
environmental conditions.

### – 84 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: Terrestrial field dissipation of BYI08330 in Florid a Soil, 2004 Author(s), year: Lenz, M., 2006b Study/Report number: M-277184-01-1, MEFNX055 Guideline(s): US-EPA No. 164-1: Terrestrial field d issipation, PMRA Data Code No. 8.3.2.1
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: Spirotetramat (unlabelled) as oil d ispersion formulation BYI08330 100 OD containing 102.02 g ai L -1 

### , batch 08030/0110(0073)
Internal standard: 13 

### C-spirotetramat, 13 

### C-spirotetramat-enol, 13 

### C-spirotetramat-ketohydroxy and 13 

### Cspirotetramat-MA-amide Plot preparation and
maintenance: Two treatments (test plots):

• Bare ground

• Cropped (bush beans ‘Blue Lake’, planted 23 rd 

### April, 2004) Both plots (each 3.0 × 36.6 m, 110 m 2 

### ) divided into three subunits (3.0 × 12.2 m, 37

m 2 

### ), one control plot (3.0 × 12.2 m, 37 m 2 

### ), bare ground plot maintained fallow with a non-selective herbicide (glyphosate) Application: 439 g ai ha -1 

### as a liquid spray, 250 L ha -1 

### , liquid broadcast spray (CO 2 

### backpack sprayer) Application date: 27 th 

### April, 2004 Study duration: 540 days Irrigation: 28 irrigation events, seasonally applie d as needed, 8 – 46 mm each time Application
verification: • Whatman filter pads (24 cm diameter, 452 cm 2 

### ), 2 pads per subunit, immediately sampled after application

• soil pans (248 cm 2 

### ) with 85 g of soil, 1 pan per subunit, immediately sampled after application Sampling: 5 cores (5.7 cm diameter) from each subpl ot per sampling data, composited into one sample per soil depth, 122 cm sampling depth, 15 cm soil core length, soil cores were analysed until two residue free layers were fo und Sample processing: • filter pads: extracted with 5 ml formic acid and 4 95 ml of 4 g L -1 

### cysteine hydrochloride in 2.5 % ethyl acetate in water/aceto nitrile (50/50, v/v) solution, aliquot fortified with internal 13 

### C standard solution, filtered and measured by
LC/MS/MS

• Soil pans: same method used for the soil samples

• Soil samples: microwave extraction (twice) of 20 g soil with 50 ml of acetonitrile/water (50/50, v/v) containing 8 g L -1 

### cysteine hydrochloride and 5 % ethyl acetate at 70 °C for 30 min, aliquot fortif ied with internal 13 

### C standard solution, filtered and measured by LC/MS/MS (analyt ical method FN-002- S05-03) Sample storage length: Soil samples stored frozen for maximum 786 days (An alysis period: 21 st 

### October
2005 – 19 th 

### June 2006) Monitoring: Daily weather conditions, soil moisture at 5 and 30 cm soil depth Analytical techniques: LC/MS/MS LOQ = 5 μg kg -1 

### (each analyte) LOD = 0.5 μg kg -1 

### (spirotetramat, spirotetramat-enol), 0.9 μg kg -1 

### (spirotetramatketohydroxy), 2.1 μg kg -1 

### (spirotetramat-MA-amide) Fortification solution: Mixture of spirotetramat, s pirotetramat-enol, spirotetramatketohydroxy and spirotetramat-MA-amide (fortificati on at 5.0 μg kg -1 

### each)

### – 85 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Kinetic evaluation: • Parent and total combined residues: simple first o rder (SFO) kinetics:

• Spirotetramat-enol and spirotetramat-ketohydroxy: Spirotetramat SFO 

###  (spirotetramat-enol + spirotetramat-ketohydroxy) SFO 

###  sink (no additional pathways to sink) (additional e valuation by the reviewer):

All data without normalization, MATLAB 7.0.4

Table B.8.1.3.1-6: Study site. Location Seminole County, Florida, USA Slope gradient: ≤ 1 % Depth to groundwater [m]: > 0.9 30 years average rainfall: 1228 mm 30 years mean temperature: 22.7 °C Weather conditions during study: Within 29 years no rmal level, total water input 134 % of average rain fall Study rainfall: 1544 mm (April 2004 – March 2005), 97 4 mm (April 2005 – October 2005) Study irrigation: 218 mm (April 2004 – March 2005), 1 06 mm (April 2005 – October 2005) Total precipitation (rainfall + irrigation): 1763 m m (April 2004 – March 2005), 1201 mm (April 2005 – Oc tober 2005) Crops grown: 2000: fallow, 2001: fallow, 2002: fall ow, 2003: sweetcorn, 2004: fallow Pesticides used: 2003: Warrior, Lambda-cyhalothrin Fertilizer used: 2003: 10-10-10, 34-0-0 Plot preparation: Tillage



### Table B.8.1.3.1-7: Soil properties (updated numbers according to the position paper Hellpointner et al., 2007). Soil depth [cm] Property 0 – 15 15 – 30 30 – 45 45 – 60 60 – 75 75 – 90 90 – 105 105 – 122 Texture (USDA) Sand Sand Sand Sand Sand Sand Sand Sand Sand (USDA) [%] 95 96 96 95 94 92 94 92 Silt (USDA) [%] 3 2 2 3 3 4 3 4 Clay (USDA) [%] 2 2 2 3 3 4 3 4 pH (soil/water, 1/1, m/v) 7.1 7.0 6.9 6.9 6.9 6.8 6 .9 6.8 pH (0.01 M CaCl 2 ) na na na na na na na na Organic carbon [%] a 0.2 0.1 0.1 0.2 0.3 0.5 0.3 0.5 CEC [meq kg -1 ] 15 10 9 14 26 42 26 42 Bulk density [g cm -3 ] 1.4 1.4 1.4 1.4 1.4 1.3 1.4 1.3 MC (1/3 bar) [%] 1.6 1.5 1.4 1.7 2.8 3.9 2.8 3.9 MC (15 bar) [%] 1.3 1.2 1.0 1.1 1.4 1.9 1.4 1.0

a Organic carbon = organic matter / 1.724



### Findings: Daily soil moisture monitored at a soil depth of 5 and 30 cm was above field capacity (at 1/3 bar) thr oughout the study duration. Therefore, sufficient water was applied to the test plot to promote leaching of th e test
substance. Parent Sink

Combined residues Sink

Parent Spirotetramat-enol +

Spirotetramat-ketohydroxy Sink

### – 86 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Mean recovery of fortified control soil samples (5. 0 and 50.0 μg kg -1 

### of each analyte) were 103, 103, 91 and 94 % for spirotetramat, spirotetramat-enol, spirote tramat-ketohydroxy and spirotetramat-MA-amide, respectively.

Bare soil plot: Application verification filter paper and soil pans showed an average parent equivalent recovery of 85 % and 95 % of the application rate, respectively. Average 0 DAT concentration of spirotetramat in the 0 – 15 cm soil segment was 37.9 μg kg -1 

### . Residues of spirotetramat were less than LOD (0.5 μg kg -1 

### ) by 14 DAT. Spirotetramat was not found below 15 cm of soil dep th with the exception of the 1 DAT sampling, where residues of spirotetramat were almost equally distr ibuted within 0 – 30 cm of soil depths (residues in the 30 – 45 cm layer < LOQ). Major transformation products w ere spirotetramat-enol and spirotetramat-ketohydrox y. Maximum of total concentration of spirotetramat-eno l was 30.3 μg kg -1 

### by 1 DAT declining to non detectable amounts by 185 DAT. The total concentration of spir otetramat-ketohydroxy reached a maximum of 46.3 μg kg -1 

### by 1 DAT, residues completely dissipated to below the LOD by 126 DAT. Residues of metabolites were not detectable > LOD below 30 cm of soil depth with the exception of the 1 DAT sampling (amounts < LOQ ).

Cropped plot: Application verification filter paper and soil pans showed an average parent equivalent recovery of 82 % and 98 % of the application rate, respectively. Average 0 DAT concentration of spirotetramat in the 0 – 15 cm soil segment was 59.1 μg kg -1 

### . Residues of spirotetramat were less than LOD (0.5 μg kg -1 

### ) by 14 DAT. Spirotetramat was not found below 15 cm of soil dep th with the exception of the 1 DAT sampling, where residues of spirotetramat were almost equally distr ibuted within 0 – 30 cm of soil depths (residues in the 30 – 45 cm layer < LOQ). Major transformation products w ere spirotetramat-enol and spirotetramat-ketohydrox y. Maximum of total concentration of spirotetramat-eno l was 36.5 μg kg -1 

### by 1 DAT declining to non detectable amounts at the end of the study. The total concentr ation of spirotetramat-ketohydroxy reached a maximu m of 47.6 μg kg -1 

### by 1 DAT, residues completely dissipated to below the LOD by 185 DAT. Residues of metabolites were not detectable > LOQ (i.e. 5 μg kg -1 

### ) below 30 cm of soil depth (few finding close to t he LOD, especially in 1 DAT samples).– 87 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.3.1-8: Average residues [μg kg-1] in soil samples (0 – 45 cm layer) collected fromFlorida bare ground plot.Spirotetramat Spirotetramat-enol Spirotetramat-ketohydroxy Spirotetramat-MA-amideDAT0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)-1< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD037.9 na na 37.928.5na na 28.5 32.6 na na 32.8 < LOD na na < LOD123.5 25.5 2.7a51.7 20.87.7 1.8a30.332.812.1 1.4a46.3< LOD < LOD < LOD < LOD39.1 < LOD < LOD 9.1 15.4 < LOD < LOD 15.438.3< LOD < LOD 38.3 < LOD < LOD < LOD < LOD71.3a< LOD < LOD 1.3 16.6 3.2a< LOD 19.8 19.7 7.2 < LOD 26.9 < LOD < LOD < LOD <LOD14< LOD < LOD < LOD < LOD 12.5 2.0a< LOD 14.5 15.0 2.7a< LOD 17.7 < LOD < LOD < LOD < LOD28< LOD < LOD < LOD < LOD 5.2 < LOD < LOD 5.2 4.1a3.8a< LOD 7.9 < LOD < LOD < LOD < LOD59< LOD < LOD < LOD < LOD 5.1 0.8a< LOD 5.9 2.3a1.4a< LOD 3.7a< LOD < LOD < LOD < LOD126< LOD < LOD < LOD < LOD 3.2 < LOD < LOD 3.2a< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD185< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD266na na na na na na na na na na na na na na na na367na na na na na na na na na na na na na na na na540na na na na na na na na na na na na na na na naaResidues < LOQ (5 μg kg-1)LOD = limit of detection (0.5 μg kg-1for spirotetramat and spirotetramat-enol, 0.9 μg kg-1for spirotetramat-ketohydroxy, 2.1 μg kg-1for spirotetramat-MA-amide)na = not analysed– 88 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.3.1-9: Total average residues [μg kg-1] in soil samples (0 – 45 cm layer) collected fromFlorida cropped plot.Spirotetramat Spirotetramat-enol Spirotetramat-ketohydroxy Spirotetramat-MA-amideDAT0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)-1< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD059.1 na na 59.133.3na na 33.3 41.8 na na 41.8 < LOD na na < LOD135.2 35.5 2.7a70.7 22.612.9 1.0ab36.530.5 17.1 < LOD47.6< LOD < LOD < LOD < LOD318.2 < LOD < LOD 18.2 16.1 < LOD < LOD 16.146.4< LOD < LOD 46.4 < LOD < LOD < LOD < LOD70.8a< LOD < LOD 0.8 8.6 8.9 0.8a17.5 19.317.7< LOD 37.0 < LOD < LOD < LOD < LOD14< LOD < LOD < LOD < LOD 7.1 2.3a< LOD 9.4 13.3 4.5a< LOD 17.8 < LOD < LOD < LOD < LOD28< LOD < LOD < LOD < LOD 6.7 < LOD < LOD 6.7 5.5 2.3a< LOD 7.8 < LOD < LOD < LOD < LOD59< LOD < LOD < LOD < LOD 3.5a1.1a< LOD 4.6a1.8a1.7a< LOD 3.5a< LOD < LOD < LOD < LOD126< LOD < LOD < LOD < LOD 3.1a1.2a< LOD 4.3a0.7a0.7a< LOD 1.4a< LOD < LOD < LOD < LOD185< LOD < LOD < LOD < LOD 1.9a1.0a< LOD 2.9a< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD266na na na na na na na na na na na na na na na na367na na na na na na na na na na na na na na na na540na na na na na na na na na na na na na na na naaResidues < LOQ (5 μg kg-1)bIn the original study determined as < LOD (n.d.).However, according to the individual replicates presented in an additional table, this value should be1.0 (mean of 3 replicates)LOD = limit of detection (0.5 μg kg-1for spirotetramat and spirotetramat-enol, 0.9 μg kg-1for spirotetramat-ketohydroxy, 2.1 μg kg-1for spirotetramat-MA-amide)na = not analysed

### – 89 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.3.1-10: Average residues [μg kg -1 

### ] of spirotetramat and parent equivalents [μg ai eq uiv kg 1 

### ] of spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amide in soil samples from 0 – 45 cm colleted from Florida bare ground and cro pped plots.
Bare ground plot Cropped plot

DAT Spirotetramat Spirotetramat -enol Spirotetramatketohydroxy SpirotetramatMAamide Total Spirotetramat Spirotetramatenol Spirotetramatketohydroxy SpirotetramatMAamide Total

-1 < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < L OD < LOD
0 37.9 35.5 38.4 < LOD 111.8 59.1 41.3 49.2 < LOD 149.6

1 51.7 35.3 54.5 < LOD 141.5 70.7 45.2 56.0 < LOD 171.9
3 9.1 19.1 45.1 < LOD 73.3 18.2 19.9 54.6 < LOD 92.7

7 1.3 24.5 31.7 < LOD 57.5 0.8 21.7 43.5 < LOD 66.0

14 < LOD 18.0 20.8 < LOD 38.8 < LOD 11.6 20.9 < LOD 32 .5
28 < LOD 6.4 9.3 < LOD 15.7 < LOD 8.3 9.2 < LOD 17.5

59 < LOD 7.3 4.4 < LOD 11.7 < LOD 5.7 4.1 < LOD 9.8
126 < LOD 4.3 < LOD < LOD 4.3 < LOD 5.3 1.6 < LOD 6.9

185 < LOD < LOD < LOD < LOD < LOD < LOD 3.6 < LOD < LOD 3.6
266 na na na na na na na na na na 367 na na na na na na na na na na 540 na na na na na na na na na na



### Table B.8.1.3.1-11: Calculated dissipation half-lif e [days] of spirotetramat, spirotetramat-enol, and spirotetramat-ketohydroxy and of total combined res idues in Florida bare ground and cropped soil plots (non-normalized) based on SFO kinetics.

Study site Substance DT 50 [days] DT 90

[days] R 2 χ 2

error [%] Formation

fraction [0 ... 1] Kinetics

Spirotetramat 0.9 2.9 0.96 2.2 - SFO Sum of spirotetramat-enol + spirotetramat-ketohydroxy b 6.6 21.9 nc 26.9 1 a All SFO in series Florida, bare ground plot Total combined residues b 7.6 25.2 0.86 22.3 - SFO

Spirotetramat 1.0 3.3 0.96 0.4 - SFO Sum of spirotetramat-enol + spirotetramat-ketohydroxy b 4.8 16.0 nc 26.9 1 a All SFO in series Florida, cropped plot

Total combined residues c 5.7 19.0 0.90 18.7 - SFO

a No additional pathways to sink assumed, therefore f ixed to 1 during modelling
b Calculated by the reviewer (see comments)

c Total combined residues: sum of spirotetramat, spi rotetramat-enol, spirotetramat-ketohydroxy, spirote tramat-MA-amide



### Conclusion: Spirotetramat dissipated rapidly under field condit ions on bare ground and cropped plots with a DT 50 

### of 0.9 and 1.0 days, respectively (SFO kinetics, R 2 

### = 0.96, χ 2 

### error ≤ 2.2 %). Respective DT 90 

### values were 2.9 and 3.3 days, respectively. Total combined residues dis sipated with a DT 50 

### of 7.6 and 5.7 days in bare ground and cropped plots, respectively. Residues of spirot etramat and its transformation products were not fo und > LOQ below the 0 – 30 cm soil layer. Combined residu es of spirotetramat-enol and spirotetramat-ketohydr oxy degraded with a DT 50 

### of 6.6 and 4.8 days in the bare ground and cropped plot, respectively.

Reviewer comments:

• Owing to the distinct storage instability of spiro tetramat-enol (degrading to spirotetramat-ketohydro xy and bound to the soil matrix during freeze storage, see Netzband, 2006), residues of spirotetramatenol and spirotetramat-ketohydroxy should not be ev aluated individually.

### – 90 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: Terrestrial field dissipation of BYI08330 in Califo rnia Soil, 2004 Author(s), year: Lenz, M., 2006c Study/Report number: M-277186-01-1, MEFNY002 Guideline(s): US-EPA No. 164-1: Terrestrial field d issipation, PMRA Data Code No. 8.3.2.1
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: Spirotetramat (unlabelled) as oil d ispersion formulation BYI08330 100 OD containing 102.02 g ai L -1 

### , batch 08030/0110(0073)
Internal standard: 13 

### C-spirotetramat, 13 

### C-spirotetramat-enol, 13 

### C-spirotetramat-ketohydroxy and 13 

### Cspirotetramat-MA-amide Plot preparation and
maintenance: Two treatments (test plots):

• Bare ground

• Cropped (tomatoes ‘Germain’s Seed’, planted 20 th 

### May, 2004) Both plots (each 90 × 6 m, approx. 540 m 2 

### ) divided into three subplots (30 × 6 m), one control plot (3 × 6 m, 18 m 2 

### ), bare ground plot maintained fallow with a nonselective herbicides (glufosinate, glyphosate) Application: 439 g ai ha -1 

### as a liquid spray, 187 L ha -1 

### , liquid broadcast spray Application date: 26 th 

### May, 2004 Study duration: 540 days Irrigation: 84 irrigation events, seasonally applie d as needed, 5 – 44 mm each time Application
verification: • Gelman solvent saturation pads (13.7 × 22 cm, 301 cm 2 

### ), 2 pads per subplot, immediately sampled after application

• soil pans (20.3 x 29.8 cm, 607 cm 2 

### ) with 650 g of soil, 1 pan per subplot, immediately sampled after application Sampling: 5 cores (5.7 cm diameter) from each subpl ot per sampling data, composited into one sample per soil depth, 122 cm sampling depth, 15 cm soil core length, soil cores were analysed until two residue free layers were fo und Sample processing: • saturation pads: extracted with 5 ml formic acid a nd 495 ml of 4 g L -1


### cysteine hydrochloride in 2.5 % ethyl acetate in wa ter/acetonitrile (50/50, v/v) solution, aliquot fortified with internal 13 

### C standard solution, filtered and measured by LC/MS/MS

• Soil pans: same method used for the soil samples

• Soil samples: microwave extraction (twice) of 20 g soil with 50 ml of acetonitrile/water (50/50, v/v) containing 8 g L -1 

### cysteine hydrochloride and 5 % ethyl acetate at 70 °C for 30 min, aliquot fortif ied with internal 13 

### C standard solution, filtered and measured by LC/MS/MS (analyt ical method FN-002- S05-03) Sample storage length: Soil samples stored frozen for maximum 778 days (an alysis period: 6 th 

### April 2006 –
13 th 

### July 2006) Monitoring: Daily weather conditions, soil moisture at 5 and 30 cm soil depth Analytical techniques: LC/MS/MS LOQ = 5 μg kg -1 

### (each analyte) LOD = 0.5 μg kg -1 

### (spirotetramat, spirotetramat-enol), 0.9 μg kg -1 

### (spirotetramatketohydroxy), 2.1 μg kg -1 

### (spirotetramat-MA-amide) Fortification solution: Mixture of spirotetramat, s pirotetramat-enol, spirotetramatketohydroxy and spirotetramat-MA-amide (fortificati on at 5.0 μg kg -1 

### each)
Kinetic evaluation: • Parent and total combined residues: simple first o rder (SFO) kinetics

### – 91 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

• Spirotetramat-enol and spirotetramat-ketohydroxy: Spirotetramat SFO 

###  (spirotetramat-enol + spirotetramat-ketohydroxy) SFO 

###  sink (no additional pathways to sink)

All data without normalization, MATLAB 7.0.4

Table B.8.1.3.1-12: Study site.
Location Fresno, California, USA Slope gradient: 0 % Depth to groundwater: 9.1 – 22.9 m 30 years average rainfall: 285 mm 30 years mean temperature: 17.3 °C Weather conditions during study: Within 30 years no rmal level, total water input 1590 % of average rai nfall Rainfall: 363 mm (May 2004 – April 2004), 43 mm (Ma y 2005 – November 2005) Irrigation: 739 mm (May 2004 – April 2004), 500 mm (May 2005 – November 2005) Total precipitation (rainfall + irrigation): 1102 m m (May 2004 – April 2004), 543 mm (May 2005 – Novemb er 2005) Crops grown: 1999: fallow, 2000: fallow, 2001: bare soil, 2002: bare soil, 2003: bare soil Pesticides used: 1999: Pyrimethanil, 2000: none, 2001 : Roundup, 2002: Roundup, 2003: Roundup
Fertilizer used: Information not available Plot preparation: Tillage



### Table B.8.1.3.1-13: Soil properties of the bare gro und test plot. Soil depth [cm] Property 0 – 15 15 – 30 30 – 45 45 – 60 60 – 75 75 – 90 90 – 105 105 – 122

Texture (USDA) Sandy loam Sandy

loam Sandy

loam Sandy

loam Sandy

loam Sandy

loam Sandy

loam Sandy

loam Sand (USDA) [%] 67 67 67 67 69 67 64 61 Silt (USDA) [%] 24 24 24 24 22 24 26 28 Clay (USDA) [%] 9 9 9 9 9 9 10 11 pH (soil/water, 1/1, m/v) 7.2 7.2 7.2 7.3 7.6 8.5 8 .5 8.7 pH (0.01 M CaCl 2 ) na na na na na na na na Organic carbon [%] a 0.29 0.23 0.17 0.17 0.12 0.12 0.12 0.12 CEC [meq kg -1 ] 57 59 59 60 62 81 96 107 Bulk density [g cm -3 ] 1.42 1.41 1.40 1.33 1.26 1.26 1.29 0.97 MC (1/3 bar) [%] 8.1 8.4 8.3 8.2 8.3 11.2 14.3 26.8

a Organic carbon = organic matter / 1.724 

### Parent Spirotetramat-enol +

Spirotetramat-ketohydroxy Sink Parent Sink

Combined residues Sink

### – 92 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.3.1-14: Soil properties of the cropped test plot. Soil depth [cm] Property 0 – 15 15 – 30 30 – 45 45 – 60 60 – 75 75 – 90 90 – 105 105 – 122

Texture (USDA) Sandy loam Sandy

loam Sandy

loam Sandy

loam Sandy

loam Sandy

loam Sandy

loam Sandy

loam Sand (USDA) [%] 71 71 71 71 71 71 69 65 Silt (USDA) [%] 22 22 22 22 22 22 27 30 Clay (USDA) [%] 7 7 7 7 7 7 4 5 pH (soil/water, 1/1, m/v) 6.8 6.9 6.9 7.0 7.3 7.3 7 .8 8.1 pH (0.01 M CaCl 2 ) na na na na na na na na Organic carbon [%] a 0.29 0.12 0.12 0.12 0.12 0.06 0.06 0.12 CEC [meq kg -1 ] 61 56 56 60 63 70 91 113 Bulk density [g cm -3 ] 1.48 1.46 1.42 1.47 1.48 1.48 1.42 1.24 MC (1/3 bar) [%] 7.6 7.7 7.2 7.0 6.9 7.4 12.3 23.1

a Organic carbon = organic matter / 1.724



### Findings: Daily soil moisture monitored at a soil depth of 5 and 30 cm were above field capacity (at 1/3 bar) th roughout the study duration. Therefore, sufficient water was applied to the test plot to promote leaching of th e test
substance.

Mean recovery of fortified control soil samples (5. 0 μg kg -1 

### of each analyte) were 98, 95, 93 and 101 % for spirotetramat, spirotetramat-enol, spirotetramat-ke tohydroxy and spirotetramat-MA-amide, respectively.

Bare soil plot: Application verification saturation paper and soil pans showed an average parent equivalent recovery o f 97 % and 132 % of the application rate, respectively. Average 0 DAT concentration of spirotetramat in the 0 – 15 cm soil segment was 100.0 μg kg -1 

### . Residues of spirotetramat were less than LOD (0.5 μg kg -1 

### ) by 14 DAT. Spirotetramat and its transformation products were not found below 15 cm of soil depth. Major transformation products were spirotetramat-enol and spirotetramat-ketohydroxy. Maximum of total concentration of spirotetramat-enol from 0 – 45 cm soil depth was 13.6 μg kg -1 

### by 2 DAT declining to non detectable amounts at the end of the study. The tot al concentration of spirotetramat-ketohydroxy reach ed a maximum of 52.6 μg kg -1 

### by 2 DAT, residues completely dissipated to below the LOD by study termination.

Cropped plot: Application verification saturation paper and soil pans showed an average parent equivalent recovery o f 89 % and 119 % of the application rate, respectively. Average 0 DAT concentration of spirotetramat in the 0 – 15 cm soil segment was 89.4 μg kg -1 

### . Residues of spirotetramat were less than LOD (0.5 μg kg -1 

### ) by 7 DAT. Spirotetramat and its transformation products were not found below 15 cm of soil depth (with the excep tion of 1 DAT sampling). Major transformation products were spirotetramat-enol and spirotetramat-ketohydroxy. Maximum of total concentration of spirotetramat-eno l from 0 – 45 cm soil depth was 9.8 μg kg -1 

### by 7 DAT declining to non detectable amounts at the end of t he study. The total concentration of spirotetramatketohydroxy reached a maximum of 43.6 μg kg -1 

### by 2 DAT, residues completely dissipated to below the LOD by 120 DAT.– 93 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.3.1-15: Total average residues [μg kg-1] in soil samples (0 – 45 cm layer) collected fromCalifornia bare ground plot.Spirotetramat Spirotetramat-enol Spirotetramat-ketohydroxy Spirotetramat-MA-amideDAT0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)-1< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD0100.0na na100.02.6ana na 2.6a9.8 na na 9.8 < LOD na na < LOD168.0 < LOD < LOD 68.0 7.0 < LOD < LOD 7.0 46.9 < LOD < LOD 46.9 < LOD < LOD < LOD < LOD223.4 < LOD < LOD 23.413.6< LOD < LOD13.6 52.6< LOD < LOD52.62.3a< LOD < LOD 2.3a70.6a< LOD < LOD 0.6a13.1 < LOD < LOD 13.1 38.9 < LOD < LOD 38.93.6a< LOD < LOD3.6a14< LOD < LOD < LOD < LOD 9.4 < LOD < LOD 9.4 22.2 <LOD < LOD 22.2 < LOD < LOD < LOD < LOD28< LOD < LOD < LOD < LOD 5.1 < LOD < LOD 5.1 11.8 <LOD < LOD 11.8 2.7a< LOD < LOD 2.7a62< LOD < LOD < LOD < LOD 2.9a< LOD < LOD 2.9a3.7a< LOD < LOD 3.7a< LOD < LOD < LOD < LOD120< LOD < LOD < LOD < LOD 0.8a< LOD < LOD 0.8a1.2a< LOD < LOD 1.2a< LOD < LOD < LOD < LOD187na na na na na na na na na na na na na na na na275na na na na na na na na na na na na na na na na357na na na na na na na na na na na na na na na na538na na na na na na na na na na na na na na na naaResidues < LOQ (5 μg kg-1)LOD = limit of detection (0.5 μg kg-1for spirotetramat and spirotetramat-enol, 0.9 μg kg-1for spirotetramat-ketohydroxy, 2.1 μg kg-1for spirotetramat-MA-amide)na = not analysed– 94 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.3.1-16: Total average residues [μg kg-1] in soil samples (0 – 45 cm layer) collected fromCalifornia cropped plot.Spirotetramat Spirotetramat-enol Spirotetramat-ketohydroxy Spirotetramat-MA-amideDAT0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)-1< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD089.4na na89.43.0ana na 3.0a5.9 na na 5.9 < LOD na na < LOD155.9 1.8a0.6a58.3 4.5a< LOD < LOD 4.5a37.7 1.1a< LOD 38.8 < LOD < LOD < LOD < LOD219.0 < LOD < LOD 19.0 6.7 < LOD < LOD 6.743.6< LOD < LOD43.6< LOD < LOD < LOD < LOD7< LOD < LOD < LOD < LOD9.8< LOD < LOD9.830.4 < LOD < LOD 30.4 4.0a< LOD < LOD 4.0a14< LOD < LOD < LOD < LOD 9.7 < LOD < LOD 9.7 27.9 <LOD < LOD 27.94.5a< LOD < LOD4.5a28< LOD < LOD < LOD < LOD 4.6a< LOD < LOD 4.6a13.6 < LOD < LOD 13.6 2.7a< LOD < LOD 2.7a62< LOD < LOD < LOD < LOD 2.3a< LOD < LOD 2.3a2.7a< LOD < LOD 2.7a< LOD < LOD < LOD < LOD120< LOD < LOD < LOD < LOD 0.9a< LOD < LOD 0.9a< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD187na na na na na na na na na na na na na na na na275na na na na na na na na na na na na na na na na357na na na na na na na na na na na na na na na na538na na na na na na na na na na na na na na na naaResidues < LOQ (5 μg kg-1)LOD = limit of detection (0.5 μg kg-1for spirotetramat and spirotetramat-enol, 0.9 μg kg-1for spirotetramat-ketohydroxy, 2.1 μg kg-1for spirotetramat-MA-amide)na = not analysed

### – 95 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.3.1-17: Average residues [μg kg -1 

### ] of spirotetramat and parent equivalents [μg ai eq uiv. kg -1 

### ] of spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amide in soil samples from 0 – 45 cm colleted from California bare ground and cropped plots.
Bare ground plots Cropped plots

DAT Spirotetramat Spirotetramatenol Spirotetramatketohydroxy SpirotetramatMAamide Total Spirotetramat Spirotetramatenol Spirotetramatketohydroxy SpirotetramatMAamide Total

-1 < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < L OD < LOD
0 100.0 3.2 11.5 < LOD 114.7 89.4 3.7 6.9 < LOD 100.0

1 68.0 8.7 55.2 < LOD 131.9 58.3 5.6 45.7 < LOD 109.6

2 23.4 16.9 61.9 2.6 104.8 19.0 8.3 51.3 < LOD 78.6

7 0.6 16.2 45.8 4.0 66.6 < LOD 12.1 35.8 4.0 51.9

14 < LOD 11.6 14.4 < LOD 26.0 < LOD 12.0 32.8 4.5 49.3
28 < LOD 6.3 13.9 3.0 23.2 < LOD 5.7 16.0 2.7 24.4

62 < LOD 3.6 4.4 < LOD 8.0 < LOD 2.8 3.2 < LOD 6.0
120 < LOD 1.0 1.4 < LOD 2.4 < LOD 1.1 < LOD < LOD 1.1

187 na na na na na na na na na na 275 na na na na na na na na na na 357 na na na na na na na na na na 538 na na na na na na na na na na



### Table B.8.1.3.1-18: Calculated dissipation half-lif e [days] of spirotetramat, combined residues of spirotetramat-enol and spirotetramat-ketohydroxy, a nd total combined residues in California bare ground and cropped soil plots (non-normalized) base d on SFO kinetics

Study site Substance DT 50 [days] DT 90

[days] R 2 χ 2

error [%] Formation

fraction [0 ... 1] Kinetics

Spirotetramat 1.0 3.5 0.98 12.1 - SFO Sum of spirotetramat-enol + spirotetramat-ketohydroxy 7.6 25.4 0.94 14.8 1 a All SFO in series California, bare ground plots Total combined residues b 8.4 27.9 0.95 11.3 - SFO

Spirotetramat 1.0 3.4 0.97 12.6 - SFO Sum of spirotetramat-enol + spirotetramat-ketohydroxy 8.7 28.9 0.90 22.4 1 a All SFO in series California, cropped plots

Total combined residues b 10.2 33.9 0.91 13.7 - SFO

a No additional pathways to sink assumed, therefore f ixed to 1 during modelling
b Total combined residues: sum of spirotetramat, spi rotetramat-enol, spirotetramat-ketohydroxy and spir otetramat-MA-amide



### Conclusion: Spirotetramat dissipated rapidly under field condit ions on bare ground and cropped plots with a DT 50 

### of 1.0 days in both systems (SFO kinetics, R 2 

### ≥ 0.97, χ 2 

### error ≤ 12.6 %). Respective DT 90 

### values were 3.5 and 3.4 days, respectively. Total combined residues dissipa ted with a DT 50 

### of 8.4 and 10.2 days in bare ground and cropped plots, respectively. Residues of spirotetra mat and its transformation products were not found > LOD below the 0 – 15 cm soil layer with the exception o f the 1 DAT sampling (cropped plots only).

Reviewer comments:

• Owing to the distinct storage instability of spiro tetramat-enol (degrading to spirotetramat-ketohydro xy and bound to the soil matrix during freeze storage, see Netzband, 2006), residues of spirotetramatenol and spirotetramat-ketohydroxy should not be ev aluated individually.

### – 96 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: Terrestrial field dissipation of BYI08330 in Washin gton Soil, 2004 Author(s), year: Lenz, M., 2006d Study/Report number: M-277189-01-1, MEFNY003 Guideline(s): US-EPA No. 164-1: Terrestrial field d issipation, PMRA Data Code No. 8.3.2.1
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: Spirotetramat (unlabelled) as oil d ispersion formulation BYI08330 100 OD containing 102.02 g ai L -1 

### , batch 08030/0110(0073)
Internal standard: 13 

### C 3 

### -spirotetramat, 13 

### C 3 

### -spirotetramat-enol, 13 

### C 3 

### -spirotetramat-ketohydroxy , 13 

### C 3 

### - spirotetramat-MA-amide Plot preparation and
maintenance: Two treatments (test plots):

• Bare ground

• Cropped (Yellow Sweet Spanish onions, planted 1 st 

### June, 2004; Sunstar Spring Wheat, planted 4 th 

### May, 2005) Both plots (each 45.7 × 18.3 m, approx. 836 m 2 

### ) divided into three subplots (15.2 × 18.3 m), one control plot (6 × 36.6 m), pesticides used during study: glyphosphate, oxyfluo rfen, bromoxynil, sethoxydim, paraquat, metribuzin, fluazifop, 2,4-D Application: 439 g ai ha -1 

### as a liquid spray, 229 L ha -1 

### , liquid broadcast spray Application date: 8 th 

### June, 2004 Study duration: 540 days Irrigation: 65 irrigation events, seasonally applie d as needed, 3 – 12 mm each time Application
verification: • Gelman saturation pads (13.7 × 22 cm, 301 cm 2 

### ), 2 pads per subplot, immediately sampled after application

• 6 soil pans (30.5 cm diameter, 779 cm 2 

### ) with 650 g of soil, 1 pan per subplot, immediately sampled after application Sampling: 5 cores (5.7 cm diameter) from each subpl ot per sampling data, composited into one sample per soil depth, 122 cm sampling depth, 15 cm soil core length, soil cores were analysed until two residue free layers were fo und Sample processing: • filter pads: extracted with 5 ml formic acid and 4 95 ml of 4 g L -1 

### cysteine hydrochloride in 2.5 % ethyl acetate in water/aceto nitrile (50/50, v/v) solution, aliquot fortified with internal 13 

### C standard solution, filtered and measured by
LC/MS/MS

• Soil pans: same method used for the soil samples

• Soil samples: microwave extraction (twice) of 20 g soil with 50 ml of acetonitrile/water (50/50, v/v) containing 8 g L -1 

### cysteine hydrochloride and 5 % ethyl acetate at 70 °C for 30 min, aliquot fortif ied with internal 13 

### C standard solution, filtered and measured by LC/MS/MS (analyt ical method FN-002- S05-03) Sample storage length: Soil samples stored frozen for maximum 752 days (an alysis period: 6 th 

### April 2006 –
28 th 

### June 2006) Monitoring: Daily weather conditions, soil moisture at 5 and 30 cm soil depth Analytical techniques: LC/MS/MS LOQ = 5 μg kg -1 

### (each analyte) LOD = 0.5 μg kg -1 

### (spirotetramat, spirotetramat-enol), 0.9 μg kg -1 

### (spirotetramatketohydroxy), 2.1 μg kg -1 

### (spirotetramat-MA-amide) Fortification solution: Mixture of spirotetramat, s pirotetramat-enol, spirotetramat-

### – 97 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

ketohydroxy and spirotetramat-MA-amide (fortificati on at 5.0 μg kg -1 

### each)
Kinetic evaluation: • Parent and total combined residues: simple first o rder (SFO) kinetics

• Spirotetramat-enol and spirotetramat-ketohydroxy: Spirotetramat SFO 

###  (spirotetramat-enol + spirotetramat-ketohydroxy) SFO 

###  sink (no additional pathways to sink)

All data without normalization, MATLAB 7.0.4

Table B.8.1.3.1-19: Study site.

Bare soil plots Cropped plots Location Ephrata, Washington, USA Slope gradient: 0 - 1 % Depth to groundwater: 22.9 ± 3 m 30 years average rainfall: 204 mm 30 years mean temperature: 11.3 °C Weather conditions during study: Temperature within 30 years normal level, total water input 878 % of average rainfall Rainfall 159 mm (June 2004 – May 2005), 81 mm (June 2005 – Nov 2005) Irrigation 640 mm (June 2004 – May 2005), 549 mm (J une 2005 – Nov 2005) Total precipitation (rainfall + irrigation) 799 mm (June 2004 – May 2005), 630 mm (June 2005 – Nov 200 5) Crops grown: 2001: wheat, potatoes, 2002: wheat, 2003: fallow, 2004: fallow 2001: Potato, 2002: Potato, 2003: Peas, 2004: Onion Pesticides used: 2001: Assure II, Gramoxone Extra, 2002: Roundup Ultra, 2003: Roundup Ultra, 2004: Fusilade, Gramoxone Extra, Goal, Touchdown, Buctril, Poast 2001: Lexone DF, Poast, Diquat, Matrix, Temik, 2002: Roundup Ultra, Gramoxone Extra, Sencor, 2003: Treflan, 2004: Goal, Touchdown, Buctril, Poast, Fusilade, Gramoxone Extra, Sencor
Fertilizer used: None None Plot preparation: Tillage, 1 st June, 2004 Tillage, 1 st June, 2004



### Table B.8.1.3.1-20: Soil properties of the bare gro und test plot. Soil depth [cm] Property 0 – 15 15 – 30 30 – 45 45 – 60 60 – 75 75 – 90 90 – 105 105 – 122

Texture (USDA) Sandy loam Loamy

sand Loamy

sand Loamy

sand Sand Sand Sand Sand

Sand (USDA) [%] 73 79 85 85 87 89 89 94 Silt (USDA) [%] 22 17 13 14 12 10 10 5 Clay (USDA) [%] 5 4 2 1 1 1 1 1 pH (soil/water, 1/1, m/v) 8.1 7.8 7.8 8.0 8.0 8.3 8 .5 8.5 pH (0.01 M CaCl 2 ) na na na na na na na na Organic carbon [%] a 0.41 0.23 0.17 0.17 0.12 0.06 0.06 0.06 CEC [meq kg -1 ] 120 124 127 118 116 114 122 104 Bulk density [g cm -3 ] 1.39 1.48 1.58 1.62 1.66 1.65 1.65 1.68 

### Parent Spirotetramat-enol +

Spirotetramat-ketohydroxy Sink Parent Sink

Combined residues Sink

### – 98 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Soil depth [cm] Property 0 – 15 15 – 30 30 – 45 45 – 60 60 – 75 75 – 90 90 – 105 105 – 122 MC (1/3 bar) [%] 12.1 10.2 8.1 7.9 6.5 6.0 5.9 4.7

a Organic carbon = organic matter / 1.724



### Table B.8.1.3.1-21: Soil properties of the cropped test plot. Soil depth [cm] Property 0 – 15 15 – 30 30 – 45 45 – 60 60 – 75 75 – 90 90 – 105 105 – 122

Texture (USDA) Loamy sand Loamy

sand Loamy

sand Loamy

sand Sand Sand Sand Sand

Sand (USDA) [%] 77 77 85 85 87 88 91 93 Silt (USDA) [%] 18 20 12 12 12 11 8 6 Clay (USDA) [%] 5 3 3 3 1 1 1 1 pH (soil/water, 1/1, m/v) 8.0 8.2 8.2 8.3 8.3 8.4 8 .6 8.7 pH (0.01 M CaCl 2 ) na na na na na na na na Organic carbon [%] a 0.35 0.17 0.17 0.17 0.12 0.06 0.06 0.00 CEC [meq kg -1 ] 121 122 135 128 113 110 105 92 Bulk density [g cm -3 ] 1.52 1.57 1.57 1.56 1.65 1.68 1.70 1.69 MC (1/3 bar) [%] 11.0 9.4 9.0 8.5 7.0 6.1 5.8 4.9

a Organic carbon = organic matter / 1.724



### Findings: Daily soil moisture monitored at a soil depth of 5 and 30 cm was above field capacity (at 1/3 bar) thr oughout the study duration. Therefore, sufficient water was applied to the test plot to promote leaching of th e test
substance.

Mean recovery of fortified control soil samples wer e 98, 98, 97 and 98 % for spirotetramat, spirotetra matenol, spirotetramat-ketohydroxy and spirotetramat-M A-amide, respectively.

Bare soil plot: Application verification filter paper and soil pans showed an average parent equivalent recovery of 98 % and 80 % of the application rate, respectively. Average 0 DAT concentration of spirotetramat in the 0 – 15 cm soil segment was 85.4 μg kg -1 

### . Residues of spirotetramat declined to less than L OD (0.5 μg kg -1 

### ) by 14 DAT. Spirotetramat and its transformation products were not found > LOD below 15 cm of soil depth. Major transformation products were spirotetramat-enol and spirotetramat-ketohydroxy. Maximum of total concentration of spirotetramat-enol was 19.3 μg kg -1 

### by 0 DAT declining to non detectable amounts by st udy termination. The total concentration of spirotetram at-ketohydroxy reached a maximum of 79.5 μg kg -1 

### by 1 DAT, residues completely dissipated to below the LO D by study termination.

Cropped plot: Application verification filter paper and soil pans showed an average parent equivalent recovery of 10 6 % and 83 % of the application rate, respectively. Ave rage 0 DAT concentration of spirotetramat in the 0 – 15 cm soil segment was 69.7 μg kg -1 

### . Residues of spirotetramat declined to less than L OD (0.5 μg kg -1 

### ) by 7 DAT. Spirotetramat and its transformation products were not found > LOD below 15 cm of soil depth. Major transformation products were spirotetramat-enol and spirotetramat-ketohydroxy. Maximum of total concentration of spirotetramat-enol was 24.1 μg kg -1 

### by 0 DAT declining to non detectable amounts at th e end of the study. The total concentration of spirot etramat-ketohydroxy reached a maximum of 84.6 μg kg -1 

### by 1 DAT, residues completely dissipated to below the LOD by 58 DAT.– 99 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.3.1-22: Total average residues [μg kg-1] in soil samples (0 – 45 cm layer) collected fromWashington bare ground plot.Spirotetramat Spirotetramat-enol Spirotetramat-ketohydroxy Spirotetramat-MA-amideDAT0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)-4< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD085.4na na85.4 19.3na na19.336.6 na na 36.6 < LOD na na < LOD128.5 < LOD < LOD 28.5 8.2 < LOD < LOD 8.279.5< LOD < LOD79.5< LOD < LOD < LOD < LOD32.1a< LOD < LOD 2.1a6.6 < LOD < LOD 6.6 71.8 < LOD < LOD 71.83.5a< LOD < LOD3.5a70.5a< LOD < LOD 0.5a4.0a< LOD < LOD 4.0a53.4 < LOD < LOD 53.4 < LOD < LOD < LOD < LOD14< LOD < LOD < LOD < LOD 2.4a< LOD < LOD 2.4a26.6 < LOD < LOD 26.6 < LOD < LOD < LOD < LOD28< LOD < LOD < LOD < LOD 1.7a< LOD < LOD 1.7a12.4 < LOD < LOD 12.4 < LOD < LOD < LOD < LOD58< LOD < LOD < LOD < LOD 0.7a< LOD < LOD 0.7a5.3 < LOD < LOD 5.3 < LOD < LOD < LOD < LOD120< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD 3.2a< LOD < LOD 3.2a< LOD < LOD < LOD < LOD175< LOD < LOD < LOD < LOD 1.2a< LOD < LOD 1.2a2.8a< LOD < LOD 2.8a< LOD < LOD < LOD < LOD269na na na na na na na na na na na na na na na na363na na na na na na na na na na na na na na na na538na na na na na na na na na na na na na na na naaResidues < LOQ (5 μg kg-1)LOD = limit of detection (0.5 μg kg-1for spirotetramat, spirotetramat-enol, 0.9 μg kg-1for spirotetramat-ketohydroxy, 2.1 μg kg-1for spirotetramat-MA-amide)na = not analysed– 100 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.3.1-23: Total average residues [μg kg-1] in soil samples (0 – 45 cm layer) collected fromWashington cropped plot.Spirotetramat Spirotetramat-enol Spirotetramat-ketohydroxy Spirotetramat-MA-amideDAT0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)0 – 15cm15 – 30cm30 – 45cmTotal(0 – 45cm)-4< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD069.7na na69.7 24.1na na24.139.0 na na 39.0 < LOD na na < LOD117.3 < LOD < LOD 17.3 8.7 < LOD < LOD 8.784.6< LOD < LOD84.6< LOD < LOD < LOD < LOD30.5a< LOD < LOD 0.5a4.7a< LOD < LOD 4.7a83.4 < LOD < LOD 83.43.0a< LOD < LOD3.0a7< LOD < LOD < LOD < LOD 3.6a< LOD < LOD 3.6a40.5 < LOD < LOD 40.5 2.3a< LOD < LOD 2.3a14< LOD < LOD < LOD < LOD 2.1a< LOD < LOD 2.1a14.2 < LOD < LOD 14.2 < LOD < LOD < LOD < LOD28< LOD < LOD < LOD < LOD 1.9a< LOD < LOD 1.9a3.2a< LOD < LOD 3.2a< LOD < LOD < LOD < LOD58< LOD < LOD < LOD < LOD 0.9a< LOD < LOD 0.9a< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD120< LOD < LOD < LOD < LOD 0.7a< LOD < LOD 0.7a< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD175< LOD < LOD < LOD < LOD 0.9a< LOD < LOD 0.9a< LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD269na na na na na na na na na na na na na na na na363na na na na na na na na na na na na na na na na538na na na na na na na na na na na na na na na naaResidues < LOQ (5 μg kg-1)LOD = limit of detection (0.5 μg kg-1for spirotetramat, spirotetramat-enol, 0.9 μg kg-1for spirotetramat-ketohydroxy, 2.1 μg kg-1for spirotetramat-MA-amide)na = not analysed

### – 101 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.3.1-24: Average residues [μg kg -1 

### ] of spirotetramat and parent equivalents of spirotetramat-enol, spirotetramat-ketohydroxy and s pirotetramat-MA-amide in soil samples from 0 – 45 cm colleted from Washington bare ground and crop ped plots.
Bare ground plot Cropped plot

DAT Spirotetramat Spirotetramatenol Spirotetramatketohydroxy SpirotetramatMAamide Total Spirotetramat Spirotetramatenol Spirotetramatketohydroxy SpirotetramatMAamide Total

-4 < LOD < LOD < LOD < LOD < LOD < LOD < LOD < LOD < L OD < LOD
0 85.4 23.9 43.1 < LOD 152.4 69.7 29.9 45.9 < LOD 145.5

1 28.5 10.2 93.5 < LOD 132.2 17.3 10.8 99.5 < LOD 127.6
3 2.1 8.2 84.5 3.9 98.7 0.5 5.8 98.1 3.3 107.7

7 0.6 5.0 62.8 < LOD 68.4 < LOD 4.5 47.7 2.6 54.8

14 < LOD 3.0 31.3 < LOD 34.3 < LOD 2.6 16.7 < LOD 19.3

28 < LOD 2.1 14.6 < LOD 16.7 < LOD 2.4 3.8 < LOD 6.2

58 < LOD 0.9 6.2 < LOD 7.1 < LOD 1.1 < LOD < LOD 1.1
120 < LOD < LOD 3.8 < LOD 3.8 < LOD 0.9 < LOD < LOD 0.9

175 < LOD 1.5 3.3 < LOD 4.8 < LOD 1.1 < LOD < LOD 1.1
269 na na na na na na na na na na 363 na na na na na na na na na na 538 na na na na na na na na na na LOD = limit of detection (0.5 μg kg -1 for spirotetramat, spirotetramat-enol, 0.9 μg kg -1 for spirotetramat-ketohydroxy, 2.1 μg kg -1 for spirotetramat-MA-amide) na = not analysed



### Table B.8.1.3.1-25: Calculated dissipation half-lif e [days] of spirotetramat, combined residues of spirotetramat-enol and spirotetramat-ketohydroxy, a nd total combined residues in Washington bare ground and cropped soil plots (non-normalized) base d on SFO kinetics.

Study site Substance DT 50 [days] DT 90

[days] R 2 χ 2

error [%] Formation

fraction [0 ... 1] Kinetics

Spirotetramat 0.4 1.4 0.99 1.6 - SFO Sum of spirotetramat-enol + spirotetramat-ketohydroxy 5.2 17.4 0.98 19.5 1 a All SFO in series Washington, bare ground plot Total combined residues a 6.3 21.0 0.98 8.5 - SFO

Spirotetramat 0.3 1.1 0.99 0.4 - SFO Sum of spirotetramat-enol + spirotetramat-ketohydroxy 4.6 15.4 0.98 7.4 1 a All SFO in series Washington, cropped plot

Total combined residues a 5.0 16.7 0.98 5.0 - SFO

a No additional pathways to sink assumed, therefore f ixed to 1 during modelling
b Total combined residues: sum of spirotetramat, spi rotetramat-enol, spirotetramat-ketohydroxy and spir otetramat-MA-amide



### Conclusion: Spirotetramat rapidly dissipated under field condit ions on bare ground and cropped plots with a DT 50 

### of 0.4 and 0.3 days, respectively (SFO kinetics, R 2 

### = 0.99, χ 2 

### error ≤ 1.6 %). Respective DT 90 

### values were 1.4 and 1.1 days, respectively. Total combined residues dis sipated with a DT 50 

### of 6.3 and 5.0 days in bare ground and cropped plots, respectively. Residues of spirot etramat and its transformation products were not fo und > LOD below the 0 – 15 cm soil layer.

Reviewer comments:

• Owing to the distinct storage instability of spiro tetramat-enol (degrading to spirotetramat-ketohydro xy and bound to the soil matrix during freeze storage, see Netzband, 2006), residues of spirotetramatenol and spirotetramat-ketohydroxy should not be ev aluated individually.

### – 102 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.3.2 Soil residue testing

This point is covered by point B.1.1.3.1.

B.8.1.3.3 Soil accumulation testing on relevant soi ls

This point is covered by point B.1.1.3.1.

B.8.1.3.4 Supplemental studies

Reference: In house laboratory validation of an analytical met hod for the determination of BYI08330 and its metabolites BYI08330-enol, BYI0 8330-ketohydroxy and BYI08330-MA-amide in soil and sediment by LC-MS/MS Author(s), year: Netzband, D. J., 2006a Study/Report number: M-277365-01-1, RAFNX012 Guideline(s): US-EPA Test Guidelines OPPTS 850-7100 (1996), OPPTS 860.1340 (1996)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substances: Spirotetramat, spirotetramat-enol, spirotetramat-ketohydroxy, spirotetramat-MAamide (all unlabelled)
Internal standard: 13 

### C 3 

### -Spirotetramat, 13 

### C 3 

### -spirotetramat-enol, 13 

### C 3 

### -spirotetramat-ketohydroxy,

13 

### C 3 

### -spirotetramat-MA-amide Test system: Two soil samples (North Rose, New York , study nr. MEFNY004, Lenz, 2006a, and Ephrata, Washington, study nr. MEFNY003, Lenz, 2006 d) and one sediment (Columbia, SC); 13 replicates for each soil, 7 repl icates for the sediment. The soils used are from the field dissipation study sites (B.8.1.3.1). The soil properties are already listed in monograph tables ( New York: Table B.8.1.3.1-2, Washington: Table B.8.1.3.1-19) A detailed sediment characterisation (SC) is not av ailable since that was not mentioned in the respective study protocol Fortification: 5 and 25 μg kg -1 

### of each analyte separately Internal laboratory
validation: An Intern laboratory validation (ILV) on the method has been performed with the soils ‘Hoefchen’ and ‘Laacherhof’, spiking level 5 and 50 μg kg -1 

### (for details refer to following study) Sample processing: Analytical method FN-002-S05-02 (updated to FN-002-S05-03): 20 g of soil are mixed with [25 mL of a solution of ethyl acetate (5 %, v/v) and cysteine hydrochloride (8 g L -1 

### ) in water, 0.5 mL formic acid, 25 mL acetonitrile] , fortified
with 1 ml of 13 

### C internal standard solution and extracted by micro wave extraction (70 °C, 30 min). Extraction (without addition of in ternal standard) is repeated once. Following extraction samples are filtered (0.45 μm) to await LC/MS/MS analysis. Analytical techniques: LC/MS/MS (screening for spir otetramat-enol dimer 1 and 2 possible)
Limit of detection (LOD): Defined as: (standard deviation (fortification 5 μg kg -1 

### ) × t 0.99 

### ) + average apparent residue in untreated control;
where t 0.99 

### is the one-tailed t-statistic at the 99 % confiden ce level of n-1 replicates (i.e. 2.68 in case of n = 13 and 3.14 in case of n = 7 )

### – 103 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Limit of quantification (LOQ): Defined as: (standard deviation (fortification 5 μg kg -1 

### ) × 5) + average apparent
residue in untreated control

Findings: Calibration solutions of each analyte are stable ov er a minimum of 4 months when refrigerated.

Table B.8.1.3.4-1: Mean recover and relative standa rd deviation (RSD) of analytes of freshly spiked soil and sediment samples.

Soil Sediment

Analyte Spiking level [μg kg -1 ] Overall mean [%] RSD [%] Overall mean [%] RSD [%]

5 101 3.6 106 3.1 Spirotetramat 25 106 1.6 108 3.2

5 93 16.9 108 16.2 Spirotetramat-enol 25 84 11.7 84 3.9

5 98 6.9 110 5.6 Spirotetramat-ketohydroxy 25 103 5.7 105 7.2

5 105 3.3 117 3.4 Spirotetramat-MA-amide 25 103 3.5 108 5.9



### Table B.8.1.3.4-2: Calculated LODs [μg kg -1 

### ] and LOQs [μg kg -1 

### ] for spirotetramat, spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amid e in soil and sediment samples.

Matrix Analyte Calculated LOD [μg kg -1 ] Calculated LOQ [μg kg -1 ]

Spirotetramat 0.5 0.9 Spirotetramat-enol 0.5 0.9 Spirotetramat-ketohydroxy 2.1 3.9 Soil

Spirotetramat-MA-amide 0.9 1.7

Spirotetramat 0.5 0.8 Spirotetramat-enol 0.6 1.0 Spirotetramat-ketohydroxy 2.7 4.4 Sediment

Spirotetramat-MA-amide 1.0 1.5



### Table B.8.1.3.4-3: Results (mean recover and relati ve standard deviation, RSD) of the ILV using the soils ‘Hoefchen’ and ‘Laacherhof’ freshly spiked wi th each analyte (overall mean of two spiking levels, 5 and 50 μg kg -1 

### ).

Hoefchen Laacherhof

Analyte Overall mean [%] RSD [%] Overall mean [%] RSD [%] Spirotetramat 94 1.8 92 1.4 Spirotetramat-enol 91 2.8 91 1.7 Spirotetramat-ketohydroxy 85 1.7 83 2.8 Spirotetramat-MA-amide 89 13.7 94 13.1



### Conclusion: The analytical method FN-002-S05-02 (updated to FN- 002-S05-03 after revision) is suitable for the determination of spirotetramat and its metabolites spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amide in soil and sediment by LC/M S/MS. The average recoveries for all analytes were
between 70 and 120 % with a relative standard devia tion (RSD) of less than 20 %. The method was shown capable of quantifying spirotetramat and its metabo lites at the targeted LOQ of 5 μg kg -1



### .

Reviewer comments:

• Details on the ILV method using the soils ‘Hoefche n’ and ‘Laacherhof’ are given in the following OLV study report.

### – 104 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: Independent laboratory validation of method FN-002- S05-02 for the
determination of BYI08330 and its metabolites BYI08 330-enol, BYI08330- keto-hydroxy and BYI08330-MA-amide in soil and sedi ment by LC/MS/MS Author(s), year: Brumhard, B., 2006 Study/Report number: M-276038-01-2, MR-06/037 Guideline(s): EC Guidance Document SANCO/825/00 rev . 7 (2004); Commission Directive 96/46/EC amending Council Dire ctive 91/414/EEC (1996); BBA Guideline: Residue Analytical Methods for Post- Registration Control Purposes (1998); US EPA Residue Chemistry Test Guidelines, OPPTS 860 .1430 (1996); US EPA Ecological Effects Test Guidelines, OPPTS 85 0.7100 (1996)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substances: Spirotetramat, spirotetramat-enol, spirotetramat-ketohydroxy, spirotetramat-MAamide (all unlabelled)
Internal standard: 13 

### C 3 

### -Spirotetramat, 13 

### C 3 

### -spirotetramat-enol, 13 

### C 3 

### -spirotetramat-ketohydroxy,

13 

### C 3 

### -spirotetramat-MA-amide Test system: • ‘Hoefchen’ soil, silt loam (USDA), 1.58 % C org



### • ‘Laacherhof’ soil, sandy loam (USDA), 2.06 % C org 

### For a detailed characterization of these soils, whi ch were also used in several laboratory studies, refer to B.1.1.1. Fortification: 5 and 50 μg kg -1 

### of each analyte separately Sample processing: Analytical method FN-002-S05-02: 20 g of soil are mixed with [25 mL of a solution of ethyl acetate (5 %, v/v) and cysteine hydrochlor ide (8 g L -1 

### ) in water, 0.5 mL formic acid, 25 mL acetonitrile], fortified with 1 ml of 13 

### C internal standard solution and extracted by microwave extraction (70 °C, 30 mi n). Extraction (without addition of internal standard) is repeated once. Fo llowing extraction samples are filtered (0.45 μm) to await LC/MS/MS analysis. Analytical techniques: LC/MS/MS LOQ: 5 μg kg -1 

### each

Findings:

Table B.8.1.3.4-4: Recovery rates of spirotetramat, spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amide.

Spirotetramat Spirotetramatenol Spirotetramatketohydroxy SpirotetramatMA-amide Fortification [μg kg -1 ] Soil Mean [%] RSD

[%] Mean

[%] RSD

[%] Mean

[%] RSD

[%] Mean

[%] RSD

[%]

5 ‘Hoefchen’ 95 2.1 93 1.2 85 2.2 78 1.6
5 ‘Laacherhof’ 93 0.7 92 1.9 85 2.9 83 2.5
5 Mean 94 1.8 92 1.8 85 2.5 80 3.9

10 ‘Hoefchen’ 93 1.1 89 1.3 85 1.2 101 1.2
10 ‘Laacherhof’ 92 1.7 91 1.5 82 2.1 106 1.9
10 Mean 92 1.7 90 1.8 83 2.1 103 3.0

Overall mean 93 1.9 91 2.3 84 2.4 92 13.3

### – 105 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion:
The obtained data demonstrate the successful valida tion of the analytical method FN-002_s05-02 for the determination of spirotetramat and its metabolites spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amide in soil. Overall mean recove ries were within a range of 80 – 94 % and thus in f ull compliance with the guideline requirements. No sign ificant changes were made to the method. The method is considered applicable as an enforcement method.

Comments (RMS):
None

Reference: Stability of BYI08330, BYI08330-enol, BYI08330-keto hydroxy and BYI08330- MA-amide in soil during frozen storage, USA, 2005 ( reported through a maximum of 334 days storage) Author(s), year: Netzband, D., 2006b Study/Report number: M-277371-01-1, RAFNX018 Guideline(s): Residue Chemistry Test Guidelines, OP PTS 860.1380 Storage Stability
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods:
Test substances: Experiment 1: Spirotetramat, spirotetramat-enol, spirotetramat-k etohydroxy and spirotetramat-MA-amide (all unlabelled) Experiment 2: [Azaspirodecenyl-3- 14 

### C]-spirotetramat-enol, purity 97.6 %,

ID No. C-1067, 2.10 MBq mL -1



### Internal standard: 13 

### C 3 

### -spirotetramat, 13 

### C 3 

### -spirotetramat-enol, 13 

### C 3 

### -spirotetramat-ketohydroxy and

13 

### C 3 

### -spirotetramat-MA-amide
Test soils: Soils from field trials:

• North Rose, New York (study nr. MEFNY004, Lenz, 20 06a)

• Oviedo, Florida (study nr. MEFNX055, Lenz, 2006b)

• Fresno, California (study nr. MEFNY002, Lenz, 2006 c)

• Ephrata, Washington (study nr. MEFNY003, Lenz, 200 6d)
Fortification: Experiment 1: 20 g soil fortified to a level of 25 μg kg -1 

### with each analyte separately Experiment 2: 5 g soil fortified with 10.4 kBq 14 

### C-spirotetramat-enol Sample set: Both experiments:

• Control samples

• Fortified samples (storage samples)

• Freshly fortified samples (for each analysis date) Storage conditions: Experiment 1: Fortified soil samples (with non-labelled test ite ms) were stored frozen at -26.8 to 1.3 °C (mean -22.7 °C) over a pe riod of 334 days Experiment 2: Fortified soil samples (with 14 

### C-spirotetrmat-enol) were stored

frozen at -21.7 to -15.8 °C (mean -19.1 °C) over a period of 20 days Sample processing: Experiment 1/Experiment 2: Soils were extracted with an acidic extraction solution in the presence of cysteine hydrochloride and utilizing microwave extraction. Samples were fortified with i nternal 13 

### C standard. Sample aliqouts were analyzed by LC/MS/MS

### – 106 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

(analytical method FN-002-S05-03) Experiment 2: Soil residues after extraction were radio-assayed by combustionLSC. Analytical techniques: Experiment 1: LC/MS/MS Experiment 2: LSC, HPLC-UV/RAD

Findings:

Experiment 1: Spirotetramat and its metabolites spirotetramat-ket ohydroxy and spirotetramat-MA-amide showed no evidence of any degradation in the four soils durin g a maximum storage interval of 334 days in frozen storage. However, reduced spirotetramat-enol recove ry was observed for each of the aged soils. Inspect ion of the results showed that significant spirotetrama t-enol loss occurred during the first month of stor age after which spirotetramat-enol loss continued at slower r ate.

Table B.8.1.3.4-5: Experiment 1: Mean recovery [% of applied] of spirotetramat, spi rotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amid e from frozen stored samples (corrected for fresh recovery).

Soil DAT Spirotetramat Spirotetramat-enol Spirotetramatketohydroxy Spirotetramat-MAamide

0 94 66 94 93
29 87 43 92 112
70 88 51 101 122 New York

334 83 37 101 107

0 105 100 84 103
30 99 67 81 118
70 109 57 80 120 Florida

329 100 24 81 106

0 102 72 74 95
28 101 45 74 127
68 97 41 76 116 California

325 101 55 75 108

0 101 93 91 95
24 101 47 89 128

71 97 32 43 116 Washington

329 104 43 83 100



### The results of the aged samples fortified with spir otetramat-enol were further examined and it was obs erved that while no spirotetramat or spirotetramat-MA-ami de residues were observed, spirotetramat-ketohydrox y was present in each of the samples. After combining spirotetramat-enol and spirotetramat-ketohydroxy (expressed as spirotetramat-enol equivalents) recov eries obtained were still lower than expected (57 – 64 % of applied by 334 DAT).

### – 107 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.3.4-6: Experiment 1: Mean recovery [μg kg -1 

### ] of spirotetramat-enol and spirotetramatketohydroxy in samples fortified with spirotetramat -enol (25 μg kg -1 

### ) and stored frozen (corrected for fresh recovery).

Spirotetramat-enol Spirotetramatketohydroxy Total Soil DAT Spirotetramat-enol equivalents [μg kg -1 ] [% of applied]

0 17.7 0.6 18.3 68
29 12.2 1.6 13.9 49
70 14.0 1.4 15.4 56 New York

334 7.6 4.9 12.6 61

0 23.2 0.3 23.5 102
30 17.5 2.0 19.5 75
70 15.5 2.4 17.9 66 Florida

329 5.2 8.7 13.9 64

0 15.7 0.9 16.6 76
28 11.9 1.7 13.6 51
68 10.8 1.6 12.4 47 California

325 12.5 1.5 13.9 62

0 20.8 0.3 21.2 95
24 11.7 3.4 15.1 61

71 8.5 3.2 11.6 44 Washington

329 9.1 2.9 12.0 57



### Experiment 2: As a supplement to these results, a shorter 14 

### C-spirotetramat-enol storage stability trial was pe rformed in order to determine the level of any un-extracted re sidues present in the soil and to determine the pre sence of any additional analytes formed by the degradation o f spirotetramat-enol while stored frozen.

The results show that the mass balance of the radio activity was in a range of 92 – 102 % of AR, by 20 DAT of frozen storage, between 3 – 15 % of AR remained bound to the soil (NER). Freshly spiked samples onl y contained spirotetramat-enol. Aged samples containe d both spirotetramat-enol and spirotetramatketohydroxy. No additional peaks were observed in t he aged samples.

Table B.8.1.3.4-7: Experiment 2: Mean recovery [% of applied] of spirotetramat-enol , spirotetramatketohydroxy, total extractable residues and non ext ractable residues in samples fortified with 14 

### Cspirotetramat-enol and stored frozen (corrected for fresh recovery).

Soil DAT Spirotetramatenol Spirotetramatketohydroxy Total Extracted Bound Total recovery

0 97 0 na na na
3 55 14 86 9 94 New York

20 43 14 85 15 100

0 86 0 na na na
3 38 40 89 3 93 Florida

20 29 44 94 3 92

0 94 0 na na na
3 52 16 90 6 93 California

20 44 14 88 8 100

0 95 0 na na na
3 47 43 87 7 96 Washington

20 19 54 92 8 102

### – 108 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion: Spirotetramat and its metabolites spirotetramat-ket ohydroxy and spirotetramat-MA-amide showed no evidence of any degradation in the four soils durin g a maximum storage interval of 334 days in frozen storage. Spirotetramat-enol recoveries significantl y declined during storage (24 – 55 % of applied by 334 DAT), with the majority of the loss occurring durin g the first 30 days of storage (43 – 67 % of applie d by 30 DAT). The primary cause of these low recoveries was degradation of spirotetramat-enol to spirotetramat

ketohydroxy (7 – 40 % equivalents of applied by 334 DAT) and binding of the analyte to soil.

Reviewer comments:

• Owing to the distinct storage instability of spiro tetramat-enol, kinetic evaluation of residues of spirotetramat-enol and spirotetramat-ketohydroxy in all US field trials is not advisable. Since it was shown, that during sample storage most of spirotetr amat-enol degrades into spirotetramatketohydroxy, it is seems more appropriate to calcul ate an overall DT 50 

### for the combined residues of spirotetramat-enol and spirotetramat-ketohydroxy.

• The submitted storage stability study report until 334 days of storage is an interim report. The storage stability studies using non-radiolabelled ( for all analytes) and radiolabelled material (only for BYI08330-enol) are ongoing. The 786 days of storage will be completed in November 2007 for the study involving non-radiolabelled material and Sept ember 2008 for the study involving radiolabelled
material.

Reference: Environmental behaviour of spirotetramat and its me tabolites in U.S. field dissipation trials Author(s), year: Röpke, B., Ramanarayanan, T., 2007 Study/Report number: -, MEF-07/120 Guideline(s): Calculations - not applicable GLP: Calculations - not applicable Deviations: Calculations - not applicable Validity: Study considered as additional information, only – study not essential

Material and methods: A kinetic data package for spirotetramat (Ramanaray anan, 2006) and its soil metabolites spirotetramat- enol, spirotetramat-ketohydroxy and spirotetramat-MA-amid e (Röpke, 2006a) was derived from laboratory aerobi c soil degradation data (Babczinski, 2005 and 2006b) for use as input parameters in environmental fate models, and in the environmental risk assessment. I n the laboratory study spirotetramat-enol showed a very rapid initial degradation phase which changed into a slow degradation after approximately one day. Thi s extremely biphasic degradation behaviour was then d escribed by a single first order reversible binding (SFORB) model. Due to this extreme biphasic degradation pattern it was not possible to derive adsorption/desorption values for spirotetramat-enol with a standard batch equilibrium study according to OECD 107. Hence, K OC 

### values for equilibrium and non-equilibrium sorptio n were derived from a column leaching study (Babczinski, 2006c). The SFO-RB kine tic model and the equilibrium sorption K OC 

### from the column leaching study were then converted into a co nsistent kinetic sorption model according to the PEARLneq (FOCUS, 2000) model which can be implement ed in environmental fate models such as PEARL.

The object of this study was to check whether the p arameter set, used in predictive environmental simu lation models, adequately describes the behaviour of spiro tetramat and metabolites under realistic outdoor conditions. Soil residue measurements of spirotetra mat and metabolites were available from four terres trial field dissipation trials conducted in the U. S. (Le nz, 2006a, 2006b, 2006c, 2006d). The terrestrial fi eld dissipation study in Florida was not used in this e xercise owing to the extreme soil and weather condi tions (extreme rainfall event in hurricane season, extrem ely light soil, 95 % sand, 0.2 % C org 

### ).

### – 109 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Flow and transport simulations for the respective f ield sites in consideration of their environmental parameters (soil, climate) were performed with FOCU S PEARL 2.2.2. Results were compared to the measured concentrations of spirotetramat and its me tabolites from the respective field studies.

Figure B.8.1.3.4-1: Multi-compartment approach (inc luding SFO-RB kinetics for spirotetramat-enol) used for modelling the behaviour of spirotetramat a nd metabolites in three US field trials (implemented into PEARL).

Spirotetramat (SFO)

Spirotetramat-enol (mobile)

Spirotetramat-ketohydroxy (SFO)

Spirotetramat-MA-amide (SFO)

Sink 100 %

26 % 74 %

100 %

100 % Spirotetramat-enol (protected) SFORB

### – 110 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.1.3.4-8: Parameter set used for modelling the behaviour of spirotetramat and metabolites in three US field trials. Compound Parameter Value Remarks

Mole mass [g mol -1 ] 373.5
Soil DT 50 [days] 0.13 SFO, normalized, geometric mean, n = 4
K FOC [L kg -1 ] 281 Batch, arithmetic mean, n = 5
1/n 0.94 Batch, arithmetic mean, n = 5 Water solubility [mg L -1 ] 29.9 20 °C, pH 7 Vapour pressure [Pa] 5.6 × 10 -9 20 °C Spirotetramat

Plant uptake 0.5

Mole mass [g mol -1 ] 301.4

Soil DT 50 [days] 0.10 SFO, normalized, geometric mean, n = 4, original value of 0.04 days substituted by the lowest limit of 0.1 days in PEARL (see comments) Formation fraction [%] (from parent) 100 Not known, conservative assessment

f ne [-] 99 Kinetic sorption parameter (SFO-RB), arithmetic mean, n

= 4, original value of 377 substituted by the upper limit of 99 in PEARL (see comments)

k d [days -1 ] 0.0159 Kinetic sorption parameter (SFO-RB), geometric mean, n

= 4
K OC [L kg -1 ] 55 Column leaching, arithmetic mean, n = 4

1/n 1.0 Set to 1.0 to adequately account for SFO-RB kinetics, conservative assessment Water solubility [mg L -1 ] 2700 20 °C, pH 7 Vapour pressure [Pa] 3.64 × 10 -10 25 °C (US EPA 2000)Spirotetramatenol

Plant uptake 0.5

Mole mass [g mol -1 ] 317.4
Soil DT 50 [days] 3.8 SFO, normalized, geometric mean, n = 4 Formation fraction [%] (from spirotetramat-enol) 26

K FOC [L kg -1 ] 63.7 Batch, arithmetic mean, n = 5
1/n 0.92 Batch, arithmetic mean, n = 5 Water solubility [mg L -1 ] 288 20 °C, pH 7 Vapour pressure [Pa] 2.21 × 10 -10 25 °C (US EPA 2000)Spirotetramatketohydroxy

Plant uptake 0.5

Mole mass [g mol -1 ] 335.4
Soil DT 50 [days] 1.0 SFO, normalized, geometric mean, n = 4 Formation fraction [%](from Spirotetramatketohydroxy) 100 Fixed to 100 % during fitting procedure

K FOC [L kg -1 ] 9.3 Batch, arithmetic mean, n = 5
1/n 0.95 Batch, arithmetic mean, n = 5 Water solubility [mg L -1 ] 69.74 20 °C, pH 7 Vapour pressure [Pa] 1.56 × 10 -11 25 °C (US EPA 2000)SpirotetramatMA-amide

Plant uptake 0.5



### Table B.8.1.3.4-9: Parameterization of soil and cli mate. Simulation domain, discretisation and boundary condi tions 122 cm soil profile, 2.5 cm intervals, free d rainage

Soil parameterization Hydraulic parameter (van Genuchten and K sat parameter) derived by ROSETTA Vegetation Bare ground and cropped plots evaluated separately

Temperature and moisture normalization FOCUS default values, Q 10 = 2.2, Walker exponent of moisture response = 0.7

### – 111 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Findings:

A

B

C

Figure B.8.1.3.4-2: Measured (squares) and modelled (lines) amounts of spirotetramat (A), of combined residues of spirotetramat-enol and spirote tramat-ketohydroxy (B) and of spirotetramatMA-amide (C) in 0 – 15 cm soil depth of the Califor nia site (bare ground plots).

### – 112 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

A

B

C

Figure B.8.1.3.4-3: Measured and modelled amount of spirotetramat (A), of combined residues of spirotetramat-enol and spirotetramat-ketohydroxy (B ) and of spirotetramat-MA-amide (C) in 0 – 15 cm soil depth of the New-York site (bare ground plots).

### – 113 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

A

B

C

Figure B.8.1.3.4-4: Measured and modelled amounts o f spirotetramat (A), of combined residues of spirotetramat-enol and spirotetramat-ketohydroxy (B ) and of spirotetramat-MA-amide (C) in 0 – 15 cm soil depth of the Washington site (bare ground plots).

### – 114 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion: The comparison of the measured soil residue of spir otetramat and its metabolites from three US field s tudies to simulated soil concentrations based on the param eter set derived by Ramanarayanan (2006) and Röpke (2006a) overall yielded good agreement between the uncalibrated model simulations and measurements. No significant residues of the active ingredient or it s metabolites below 15 cm of depth were predicted b y the model and this was confirmed by the field study dat a. In general it can be concluded that the environm ental behaviour of spirotetramat and its metabolites unde r field conditions is adequately and rather conserv atively described by the environmental fate (adsorption and degradation) data and kinetic sorption model which was derived from laboratory studies and used in the env ironmental exposure assessment for the EU calculati ons.

Reviewer comments:

• Study not considered essential. Study results included for additional information o nly.

• Owing to the low stability of spirotetramat-enol e ven under frozen storage conditions (degrading to spirotetramat-ketohydroxy and formation of NER), mo delled data for spirotetramat-enol and spirotetramat-ketohydroxy can be only compared to t he combined residues of these two metabolites
measured in the field trials.

• Owing to numerical restrictions of the PEARL model , no longer DT 50 

### values than 0.1 days (lower limit) and f ne 

### values of 99 (upper limit) can be used as substitu tes in PEARL modelling. With this restricted parameter set as used in PEARL leaching assessments, the formation of spirotetramatketohydroxy is underpredicted by approx. 21 % on av erage. An increase in the formation fraction of spirotetramat-ketohydroxy by this magnitude is not expected to have a significant impact on the groundwater risk assessment (position paper, Hellpo intner et al., 2007).

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.1.3.5 Summary on field dissipation studies

Four field dissipation studies (including bare and cropped soils) were conducted with spirotetramat on a range of soil types in the USA, representing rather worst-case scenarios in respect to degradation (lo w amounts of organic C) and leaching behaviour (sand y soils with low contents of clay). Spirotetramat w as applied once at 439 g ai ha -1 

### (single application) formulated as the oil dispers ion BYI08330 100 OD.

Dissipation of spirotetramat in field trials was ra pid with a dissipation half-life (DT 50 

### ) in a range of 0.3 – 1.0 day (SFO kinetics), respective DT 90 

### values were in a range of 1.1 – 3.5 days. Since al most no transfer of spirotetramat into soil layers below 15 cm was obse rved and volatilization is considered to be minimal (vapour pressure 5.6 × 10 -9 

### Pa), dissipation of spirotetramat is considered mo re or less consistent with degradation. No differences were observed between b are ground and cropped plots. Owing to the short ha lflife of spirotetramat in soil, potential of accumul ation and carry over of spirotetramat from one year to the other year is considered negligible.

Quantification of metabolite residues in the field trial was strongly affected by the low storage stability of spirotetramat-enol . In soil storage experiments (spiked soil samples, stored over a time period of 334 days, using the same soils from the four field trials), i t was demonstrated that spirotetramat-enol extensiv ely degrades to spirotetramat-ketohydroxy and is also s trongly bound to the soil (NER) even under frozen storage conditions. Decline of spirotetramat-enol w as most pronounced during the first 30 days of stor age. In this respect, residues of spirotetramat-enol found in the field trials (including sample storage time of several months) are likely to be significantly underestimat ed, whereas residues of spirotetramat-ketohydroxy a re vice

versa overestimated. Therefore, an individual kinetic ev aluation of spirotetramat-enol and spirotetramatketohydroxy residues measured in the field trials i s not advisable at all. Spirotetramat, spirotetrama tketohydroxy and spirotetramat-MA-amide are consider ed stable under conditions of frozen storage.

Based on combined residues of spirotetramat-enol an d spirotetramat-ketohydroxy, DT 50 

### values in a range of 4.6 – 31.6 days were found (SFO kinetics). Respecti ve DT 90 

### values were in a range of 15.4 – 105 days. No significant differences were found between bare gro und and cropped plots.

Total combined residues (spirotetramat, spirotetram at-enol, spirotetramat-ketohydroxy and spirotetrama tMA-amide) were found to dissipate with a DT 50 

### in a range of 5.0 – 23.4 days (SFO kinetics), resp ective DT 90 

### values were in a range of 16.7 – 77.8 days. No diff erences between plot treatments were found at all. Potential of carry over of any residue from one yea r to the other year is considered minimal.

Residues of spirotetramat did not move below the su rface layer (0 – 15 cm) in all sites, except Florid a where residues of spirotetramat-enol and spirotetramat-ke tohydroxy were detected above the LOQ (i.e. 5 μg kg -1 

### ) at 15 – 30 cm layer between 1 and 7 DAT. Thereafter, r esidues completely degraded to less than LOQ and
LOD. It should be noted that the Florida test site represents worst-case conditions with heavy rainfal l, very light soil (95 % sand in the top layer) and low org anic matter (0.2 %). Based on field dissipation dat a, leaching and groundwater contamination is not likel y to occur after application of spirotetramat accor ding to
GAP.

In order to validate degradation and kinetic adsorp tion/desorption parameters of spirotetramat and its metabolites obtained in laboratory experiments, the US field trials conducted in California, New York and Washington were fully re-modelled with the identica l parameter set as suggested for the groundwater ri sk assessment (implementing SFO-RB kinetics for spirot etramat-enol). From the close fit between measured and modelled data (uncalibrated model based on labo ratory data), it can be concluded that the adsorption/desorption and degradation parameter set , suggested for the environmental risk assessment,

### – 116 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

adequately reflects the transport and degradation b ehaviour of spirotetramat and its metabolites under real
field conditions.– 117 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.1.3.5-1: Summary on dissipation half-lives of spirotetramat in four US dissipation field trials (non-normalized data).TexturepH(CaCl2)pH(soil/water,1/1, m/v)Organic C[%]Clay[%]Site Crop0 – 15 cmDT50[days]DT90[days]R2χ2error[%]Kinetics‘New York’, US Bare ground Loamy sand 5.5 6.3 0.709 0.5 1.6 0.94 5.1 SFOBare ground Sand - 7.1 0.23 2 0.9 2.9 0.96 2.2 SFO‘Florida’, USBush beans Sand - 7.1 0.23 2 1.0 3.3 0.96 0.4 SFOBare ground Sandy loam - 7.2 0.29 9 1.0 3.5 0.98 12.1 SFO‘California’, USTomatoes Sandy loam - 6.8 0.29 7 1.0 3.4 0.97 12.6SFOBare ground Sandy loam - 8.1 0.41 5 0.4 1.4 0.99 1.6 SFO‘Washington’, USOnions Loamy sand - 8.0 0.35 5 0.3 1.1 0.99 0.4 SFOGeometric mean (EU)0.72.2---80th%ile (PMRA)1.03.4---90th%ile of C.I. on meana(US-EPA)0.93.0---aCalculated by [arithmetic mean + (t90,n-1× SD) / n0.5], wherebyt90,n-1= one-sided studentstvalue atα= 0.1, SD = standard deviation and n = number of data availableTable B.8.1.3.5-2: Summary on dissipation half-lives of combined residues of spirotetramat-enol and spirotetramat-ketohydroxy and of total combinedresidues in four US dissipation field trials (0 – 15 cm, non-normalized data).Sum of spirotetramat-enol and spirotetramat-ketohydroxy Total combined residuesSite CropDT50[days] DT90[days]χ2error [%]R2DT50[days] DT90[days]χ2error [%]R2‘New York’, US Bare ground 31.6 105 16.6 nc 23.4 77.8 16.4 0.93Bare ground 6.6 21.9 26.9 nc 7.6 25.3 22.3 0.86‘Florida’, USBush beans 4.8 16.0 26.9 nc 5.7 19.0 18.7 0.90Bare ground 7.6 25.4 14.8 0.94 8.4 27.9 11.3 0.95‘California’, USTomatoes 8.7 28.9 22.4 0.90 10.2 33.9 13.7 0.91Bare ground 5.2 17.4 19.5 0.98 6.3 21.0 8.5 0.98‘Washington’, USOnions 4.6 15.4 7.4 0.98 5.0 16.7 5.0 0.98Geometric mean (EU)7.725.6--8.327.7--80th%ile (PMRA)8.528.2--9.832.7--90th%ile of C.I. on meana(US-EPA)15.250.4--13.043.2--nc denotes not calculatedaCalculated by [arithmetic mean + (t90,n-1× SD) / n0.5], wherebyt90,n-1= one-sided studentstvalue atα= 0.1, SD = standard deviation and n = number of data available

### – 118 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.2 Adsorption, desorption and mobility in soil

B.8.2.1 Mobility studies (OECD Annex IIA, 7.4, IIIA , 9.3)

B.8.2.1.1 Adsorption and desorption of the active s ubstance (OECD Annex IIA, 7.4.1)

The adsorption/desorption of spirotetramat was inve stigated in one study using [3- 14 

### C] labelled parent.

Reference: BYI08330: Adsorption/desorption in five soils Author(s), year: Haas, M., 2005 Study/report number: M1311388-5, MEF-04/373 Guideline(s): OECD Guidelines 106 (2000), US-EPA N 163-1 (1982), Canadian PMRA DACO 8.2.4.2 (1987) 1 

### , Japanese MAFF New Test Guidelines for Supporting Registration of Pesticides (2000 and 2001)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: [Azaspirodecenyl-3- 14 

### C]-spirotetramat, 3.67 MBq mg -1



### , > 98 % radiochemical purity (HPLC), batch BECH 1537 and 1597 Reference substance: Spirotetramat (unlabelled), sp irotetramat-enol, spirotetramat-ketohydroxy Soils: Air dried, 2-mm sieved Soil/solution ratio: 1/10, m/v Temperature: 20 ± 1 °C Test concentrations: 0.01, 0.03, 0.1, 0.3 and 1.0 m g L -1



### in 0.01 M CaCl 2 

### (containing 50 mg L -1 

### mercuric(II)chloride for sterilization)
Co-solvent: None Pre-equilibration time: 48 hrs Equilibration time: 3 hrs for adsorption, 3 hrs for desorption (highest test concentration: 3 desorpti on cycles) Extraction: After adsorption remaining soil was ext racted twice with 20 ml acetonitrile/water (1/1, v/v). After extraction the soil residues were radio-assayed by combustionLSC. Analytical techniques: LSC, HPLC-UV/RAD, HPLC-MS/MS (ESI)
LOQ = 1 % of AR Calculations: The amount adsorbed was calculated by subtracting the equilibrium concentration in the solution from the initial concentration appl ied. The Freundlich adsorption isotherm is described as C s 

### = K F(ads) 

### × C w (1/n) 

### , whereby
C s 

### is the equilibrium concentration of the test item in the soil after adsorption [μg g -1 

### ], C w 

### the equilibrium concentration in the solution [μg mL -1 

### ], K F(ads) 

### is the Freundlich adsorption coefficient and 1/n the Freun dlich exponent. K F(ads) 

### and 1/n were obtained by plotting log(C s 

### ) vs. log(C w 

### ) to give 1/n as the slope and log(K F(ads) 

### ) as the intercept of the linear regression. Desorp tion (based on the desorption constant K F(des) 

### ) was calculated in a similar way. In addition, the linear adsorption coefficient K d(ads) 

### was obtained by linear regression plotting C W 

### vs. C S 

### without data transformation (calculated by the reviewer).

1 The Canadian guideline is improperly cited in the study report. It should be cited as: Agriculture Can ada (1987) Trade Memorandum T-1-255, Guidelines for Determining Envir onmental Chemistry and Fate of Pesticides.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Based on the organic carbon [%], the Freundlich ads orption coefficient is expressed as K FOC(ads) 

### = (K F(ads) 

### × 100) / C org 

### (Freundlich desorption and linear adsorption/desorption coefficients are expressed in a similar way).

Table B.8.2.1.1-1: Physico-chemical characteristics of test soils used for adsorption/desorption of spirotetramat.

Soil ‘Laacherhof

AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

Batch ID 20040122 20030729 20040109 20030324 980803

Geographic location Monheim/NorthrhineWestfalia/ Germany Monheim/

NorthrhineWestfalia/ Germany Burscheid/

NorthrhineWestfalia/ Germany Pensacola

Florida/ USA Saskatchewan/

Canada

Texture (USDA) Loamy sand Sandy loam Silt loam Sandy loa m Loam

Soil taxonomic classification (USDA) Sandy, mixed, mesic Typic Cambudolls Loamy, mixed,

mesic Typic Argudalfs Inseptic

Hapludalf Fine-loamy,

kaolinitic, thermic, Plinthic Kandiudults Orthic dark brown

Sand (USDA) [%] 83.0 52.5 19.0 77.3 27.3 Silt (USDA) [%] 10.0 30.9 66.0 12.7 48.3 Clay (USDA) [%] 7.0 16.7 15.0 10.0 24.4 pH (CaCl 2 ) 6.1 6.8 5.9 5.4 4.7 pH (water) 6.5 7.4 6.3 6.1 5.2 pH (KCl) 6.2 6.9 5.8 nd nd Organic carbon [%] 2.38 0.87 2.33 0.93 2.33 CEC [meq kg -1 ] 110 48 129 60 199 WHC (0.33 bar) [%] 12.6 20.3 21.4 18.4 nd MWHC [%] 34.4 37.3 63.1 nd nd



### Findings: In the pre-test, it was obvious that spirotetramat rapidly degrades to spirotetramat-enol in the prese nce of soil. Hence, to prevent microbial degradation of sp irotetramat the test soils were sterilized by soil equilibration in 0.01 M CaCl 2 

### containing 50 mg L -1 

### mercuric(II)chloride for two days. Under sterile c onditions (preliminary parental mass balance test), spirotetr amat was stable for 3 hrs of adsorption equilibrium time in all soils (test item recovery: 91.8 – 93.7 % of AR) with the exception of the ‘Hoefchen’ soil, where a pprox. 10 % of the test-item was degraded into spirotetramat- enol. The test item did not adsorb to the wall surf ace of
the test vessel.

The overall material balance for all concentrations for individual samples was in the range of 72.0 – 125.1 %
of AR with a mean of 97.6 % of AR. After 3 hrs of e quilibration 17.2 – 46.7 % of AR was adsorbed to th e test soils. The calculated adsorption constants K F(ads) 

### of the Freundlich isotherms for the five test soil s were in a range of 3.70 – 4.79 L kg -1 

### , the Freundlich exponent 1/n was in a range of 0.8 23 – 1.042. The desorption
K F(des) 

### values were 4 – 8 times higher than those obtained for adsorption indicating a strong binding of the
test substance once adsorbed to the soil.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.2.1.1-2: Preliminary test I: Equilibratio n test adsorption . Adsorption of the test-item after a shaking period of 0 – 24 hrs (expressed as [% of AR ] remaining in the solution and purity [% ai]), soil/solution ration 1/10 (m/v), test concentration 1.08 mg L -1 

### , containing 50 mg L -1 

### mercuric(II)chloride for sterilization.

‘Laacherhof AXXa’ ‘Laacherhof AIII’ ‘Hoefchen’ ‘Mol ino’ ‘Saskatoon’ HAT [% of AR] [% ai] [% of AR] [% ai] [% of AR] [% ai] [% of AR] [% ai] [% of AR] [% ai]

0 74.1 100.0 78.1 100.0 69.8 100.0 79.2 100.0 64.9 10 0.0
0.5 72.9 100.0 74.9 100.0 65.7 97.5 76.9 100.0 62.6 100 .0

1 72.9 100.0 68.7 100.0 64.6 91.0 75.2 100.0 57.9 100 .0
3 66.1 98.0 70.8 100.0 66.4 88.0 76.1 100.0 58.1 100. 0
6 71.4 100.0 73.0 100.0 70.7 79.9 77.9 95.4 68.6 89.5
24 58.7 61.7 69.2 100.0 65.0 36.9 71.2 90.4 65.0 67.5



### Table B.8.2.1.1-3: Parental mass balance . Recover of test item [% of AR] in soil after prel iminary equilibrium test adsorption and extraction (3 hrs e quilibrium time, soil/solution ration 1/10 (m/v), t est concentration 1.08 mg L -1 

### , containing 50 mg L -1 

### mercuric(II)chloride for sterilization).

Compartment ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

Solution 64.8 70.8 58.4 76.1 58.1
ACN/water extract 1 23.9 16.4 27.4 15.6 21.5
ACN/water extract 2 3.1 5.5 1.7 2.0 12.3 Test item

Recovery 91.8 92.7 87.5 93.7 91.9

Unidentified in solution 1.3 a 0.0 8.0 a 0.0 0.0
Unidentified in ACN/water extracts 0.0 0.0 0.0 0.0 0 .0
NER 1.7 2.8 1.8 0.5 2.5 Total recovery 94.8 95.5 97.3 94.2 94.4

a Mostly spirotetramat-enol– 121 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.2.1.1-4: Calculated non-linear (Freundlich) and linear adsorptionisotherm parameter for spirotetramat.Non-linear isotherm (Freundlich) Linear isothermSoil TexturepH(CaCl2)pH (sup.)aCorg[%]KF(ads)[L kg-1]KFOC(ads)[L kg-1]1/n R2Kd(ads)[L kg-1]KOC(ads)[L kg-1]R2‘Laacherhof AXXa’ Loamy sand 6.1 6.6 2.38 4.79 2011.001 0.980 4.38 184 0.988‘Laacherhof AIII’ Sandy loam 6.8 6.2 0.87 3.78 4350.892 0.993 3.80 437 0.972‘Hoefchen’ Silt loam 5.9 6.9 2.33 4.10 176 0.945 0.994 4.69 201 0.997‘Molino’ Sandy loam 5.4 6.6 0.93 4.05 435 0.823 0.976 3.58 385 0.809‘Saskatoon’ Loam 4.7 6.3 2.33 3.70 159 1.042 0.9035.52 237 0.914apH in the supernatant of the test solution.Table B.8.2.1.1-5: Calculated Freundlich desorptionisotherm parameter for spirotetramat.1stdesorptionSerial desorption(only highest test concentration)bSoil TexturepH(CaCl2)pH(sup.)aCorg[%]KF(des)[L kg-1]KFOC(des)[L kg-1]1/n R2KFser[L kg-1]KFOCser[L kg-1]1/n‘Laacherhof AXXa’ Loamy sand 6.1 6.6 2.38 40.7 17101.207 0.900 3.03 127 0.482‘Laacherhof AIII’ Sandy loam 6.8 6.2 0.87 22.8 26190.985 0.988 2.69 310 0.430‘Hoefchen’ Silt loam 5.9 6.9 2.33 14.2 610 0.952 0.986 3.19 137 0.504‘Molino’ Sandy loam 5.4 6.6 0.93 33.6 3616 0.949 0.997 2.44 263 0.388‘Saskatoon’ Loam 4.7 6.3 2.33 23.0 988 1.315 0.9234.18 179 0.621apH in the supernatant of the test solutionbCalculated by the reviewer

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion: The adsorption constants K F(ads) 

### of spirotetramat calculated on the basis of the Fr eundlich isotherms for the five test soils ranged from 3.70 – 4.79 L kg -1 

### . Respective calculated K FOC(ads) 

### values were in the range of 159 – 435 L kg -1 

### , 1/n values were in a range of 0.89 – 1.04. Spirot etramat is classified as medium mobile according to the classification scheme of McCall et al. (1981) 1 

### and moderately mobile according to FAO 2



### classification.

Reviewer comments:

• Linear adsorption constants (K OC(ads) 

### ) were close to the Freundlich adsorption constants with a range of 184 – 437 L kg -1



### .

1 McCall, J.P., D.A. Laskowsky, R.L. Swann, and H.J. D ishburger. 1981. Measurement of sorption coefficien ts of organic chemicals and their use in environmental fate analy sis. Pages 89-109 In Test protocols for environmental fate & movement of toxicants. Proceedings of a symposium. Association of Official Analytical Chemists. 94 th Annual Meeting, October 21-22, 1980. Washington, DC.
2 Food and Agriculture Organization of the United Na tions. FAO PESTICIDE DISPOSAL SERIES 8. Assessing Soil Contamination: A Reference Manual. Appendix 2. Paramet ers of pesticides that influence processes in the s oil. Editorial Group, FAO Information Division: Rome, 2000.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.2.1.2 Adsorption and desorption of relevant met abolites, degradation and reaction products

(OECD Annex IIA, 7.4.2)

The adsorption/desorption behaviour of the followin g metabolites of spirotetramat was investigated:

• Spirotetramat-enol (batch equilibrium experiment, aged adsorption study)

• Spirotetramat-ketohydroxy (batch equilibrium exper iment)

• Spirotetramat-MA-amide (batch equilibrium experime nt)

• Spirotetramat-enol dimer 1 and dimer 2 (HPLC metho d)

Reference: Adsorption/desorption of BYI08330-cis-Enol in five different soils Author(s), year: Hein, W., 2005a Study/Report number: M-267858-01-2, IM2000 Guideline(s): 95/36/EC (1995), OECD Guidelines 106 (2000), US-EPA N 163-1 (1982), Canadian PMRA DACO 8.2.4.2 (1987) 1



### GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: [Azaspirodecenyl-3- 14 

### C]-spirotetramat-enol, 4.54 MBq mg -1



### , > 98 % radiochemical purity (HPLC), batch BECH 0917 and 1626 Reference substance: Spirotetramat-enol (unlabelled ), spirotetramat-ketohydroxy Soils: Air dried, 2-mm sieved Soil/solution ratio: 1/1, m/v Temperature: 20 ± 1 °C Test concentrations: No definitive test conducted, test concentrations used for preliminary test was 1.0 mg L -1 

### in 0.01 M CaCl 2 

### (either with or without 50 mg L -1 

### mercuric(II)chloride as biocide)
Co-solvent: None Preliminary tests: Preliminary tests were conducted to ensure appropriate soil/solution ratio, appropriate equilibrium time, sorption to the test vessel and stability of the test
item in the test solution Pre-equilibration time 16 hrs Equilibration time: No definitive test conducted ow ing to instability of test item in preliminary test s Sample processing: After phase separation, soil sam ples were extracted twice with 40 ml of acetonitrile/water (1/1, v/v, adjusted to approx. p H 9 with ammonium hydroxide, additional buffered by the addition of 1 g L -1 

### ammonium chloride). Extracts were combined, radio-assayed by LSC and analysed by HPLC

. Analytical techniques: LSC, HPLC-UV/RAD Calculations: No definitive test conducted owing to instability of test item in preliminary tests

1 The Canadian guideline is improperly cited in the study report. It should be cited as: Agriculture Can ada (1987) Trade Memorandum T-1-255, Guidelines for Determining Envir onmental Chemistry and Fate of Pesticides.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.2.1.2-1: Physico-chemical characteristics of test soils used for adsorption/desorption of spirotetramat-enol.

Soil ‘Laacherhof

AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

Batch ID 20020822 20020822 20040607 81704-S 980803

Geographic location Monheim/NorthrhineWestfalia/ Germany Monheim/

NorthrhineWestfalia/ Germany Burscheid/

NorthrhineWestfalia/ Germany Pensacola

Florida/ USA Saskatchewan/

Canada

Texture (USDA) Sandy loam Silt loam Silt loam Sandy loam Loam Sand (USDA) [%] 72.4 36.9 12.7 76 27.3 Silt (USDA) [%] 22.6 51.1 72.5 16 48.3 Clay (USDA) [%] 5.0 12.0 14.7 8 24.4 pH (CaCl 2 ) 6.3 7.6 6.1 6.2 4.7 pH (water) 6.9 6.8 6.7 6.3 5.2 pH (KCl) 6.4 nd 6.4 5.4 nd Organic carbon [%] 1.47 0.88 2.62 0.87 2.3 CEC [meq kg -1 ] 103 98 160 50 199 WHC (0.33 bar) [%] 29.1 20.7 31.3 7.4 nd MWHC [%] 34.4 36.4 72.7 23.0 nd



### Findings: Spirotetramat-enol was found to be stable in the 0. 01 M CaCl 2 

### solution over a time period of 48 hrs, no adsorption to the test vessel was observed.

From preliminary soil/solution ratio test (‘Hoefche n am Hohenseh 4a’ and ‘Molino’ soil), it was obviou s that spirotetramat-enol was not stable after an adsorpti on phase of 24 hours, neither without nor with a bi ocide. Adsorption rate was in a range of 17.1 – 69.9 %, in the supernatant spirotetramat-enol accounted for 3 6.1 – 90.5 % of total peak areas depending on the soil/so lution ratio. Investigating the adsorption equilibr ium at a soil ratio of 1/2 (m/v), no plateau concentration c ould be observed within 48 hrs. The rapid disappear ance of the test item during the test can be mainly explain ed by transformation to spirotetramat-ketohydroxy ( and other degradation products) and strong binding to t he soil matrix. With parent mass balance below 90 % the quality criteria as set out in the current OECD gui deline could not be met.

Table B.8.2.1.2-2: Total recovery of radioactivity and spirotetramat-enol mass balance for preliminary equilibrium tests on adsorption in the absence and presence of biocide. Total AR Spirotetramat-enol

Soil Biocide DAT Supernatant Extract Soil residues (NER) Total Supernatant Extract Total

0.25 70.7 6.7 20.4 97.8 67.2 5.6 72.8

1 30.6 14.8 51.8 97.2 20.6 7.1 27.7 Without biocide 2 21.1 14.8 60.6 96.4 8.1 5.9 14.0

0.25 73.7 7.0 17.6 98.3 69.7 6.2 75.8

1 48.7 13.8 35.4 97.8 37.5 8.8 46.3 ‘Hoefchen’

With

biocide 2 36.8 16.5 41.5 94.7 23.7 8.9 32.6

0.25 82.0 9.4 7.5 98.8 79.6 8.9 88.5

1 55.8 17.3 25.0 98.1 50.2 12.1 62.3 Without biocide 2 31.3 20.8 46.1 98.1 20.7 9.7 30.4

0.25 78.3 11.4 9.9 99.5 75.5 10.4 86.0

1 57.4 18.8 20.5 96.7 50.0 12.2 62.2 ‘Molino’

With

biocide 2 45.5 22.9 29.6 97.9 35.0 12.3 47.2

### – 125 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion: No definitive test data regarding the adsorption/de sorption characteristics of spirotetramat-enol coul d be derived within this study. The rapid disappearance of spirotetramat-enol during preliminary test can b e mainly attributed to transformation to spirotetrama t-ketohydroxy (and other degradation products) and strong binding to the soil matrix. With parent mass balanc e below 90 % the quality criteria as set out in the current OECD guideline could not be met.

Reviewer comments:
None

Reference: [Azaspirodecenyl-3-14C]BYI08330-enol: Time-dependen t sorption in soils Author(s), year: Menke, U., 2006b Study/report number: M131 1375-1, MEF-05/222 Guideline(s): Supportive non-guideline study, based on parts of OECD Guideline 106 (2001) and US-EPA N 163-1 (1982)
GLP: Yes Deviations: Supportive non-guideline study - not ap plicable Validity: Study considered as additional information, only – study not essential

Material and methods: Test substance: [Azaspirodecenyl-3- 14 

### C]-spirotetramat-enol, 4.54 MBq mg -1



### , 96.8 % radiochemical purity (HPLC), batch BECH 1539 Reference substances: Spirotetramat-enol (unlabelle d), spirotetramat, spirotetramat-ketohydroxy, spirotetramat-MA-amide Application: 20 g d.w. soil (air dried, approx. 25 – 30 % of MWHC) were fortified with spirotetramat-enol (in 1.2 mL of 0.01 M NaHCO 3 

### solution) at a spike level of 310 μg kg -1 

### (brought to a MWHC of 50 % with distilled water). Co-solvent: 0.22 % of methanol in application solut ion Aging: After application of the test item the spike d samples were incubated for 0, 1, 5 and
24 hrs at 20 ± 1 °C in the dark at 50 % of MWHC. Desorption: After aging, spiked soil samples were o verlaid with 20 ml of 0.01 M CaCl 2 

### solution resulting in a soil to solution ratio of 1/1 (m/v) and shaken for 0.03, 1.03, 3, 5 and 24 hrs. After phase separation, supernatants were r adio-assayed by LSC and analysed by HPLC. Extraction procedure: After each desorption period, soil samples were extracted three times with acetonitrile/water/ammonia/ammonium chloride (500/5 00/2.5/1.0, v/v/v/w), extracts were combined and radio-assayed by LSC. So il residues were additionally extracted with the same solvent using a microwave at 70 °C and the extract radio-assayed by LSC. Both extract were fur ther analysed by HPLC. Non extractable residues (NER) were analysed by combust ion-LSC. Analytical techniques: LSC, HPLC-UV/RAD, LC-MS/MS
LOQ ≤ 0.4 % of AR
Calculations: Amounts of ‘test item adsorbed to the soil’ (C extr. 

### ) are calculated from the sum of residue amounts in the organic and aggressive extra cts.

### – 126 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.2.1.2-3: Soils used for adsorption/desorp tion of spirotetramat.

Soil ‘Hoefchen’ ‘Molino’

Batch ID 20040607 20040427

Geographic location Burscheid/Northrhine Westfalia/ Germany Pensacola/Florida/ USA Texture (USDA) Silt loam Sandy loam Sand (USDA) [%] 15.6 77.3 Silt (USDA) [%] 68.1 12.7 Clay (USDA) [%] 16.2 10.0 pH (CaCl 2 ) 6.1 5.4 pH (water) 6.7 6.0 pH (KCl) 6.4 5.4 Organic carbon [%] 2.62 0.8 CEC [meq kg -1 ] 160 42 MWHC [%] 67.8 26.3



### Findings:
The test item was stable in 0.01 M CaCl 2 

### solution over a period of 24 hrs (93.5 % of test i tem recovered), no indications for adsorption to the surface of the te st vessels was observed.

The total recovery of radioactivity was in a range of 91.1 – 98.9 % of AR for ‘Hoefchen’ soil and 96.0 – 99.0
% of AR for ‘Molino’ soil. Formation of NER was a s trong time-depending process even in samples withou t aging period and was increasing with increasing tim e of desorption. In non-aged samples of ‘Hoefchen’ soil
NER increased from 19.8 to 53.7 % of AR from 0.03 t o 24 hrs of desorption phase, in of ‘Molino’ soil a similar increase was observed (13.7 to 49.6 % of AR).

The instability of spirotetramat-enol in the test s ystem once in contact with soil was pronounced, aft er an aging period of 24 hrs and a desorption phase of 24 hrs only 17.5 and 18.6 % of AR could be recovered in the extracts of ‘Hoefchen’ and ‘Molino’ soil, respe ctively. The amount of total radioactivity desorbed by the aqueous CaCl 2 

### solution rapidly decreased with time of aging from 52.0 to 5.4 % of AR and 55.1 to 4.8 % of AR in ‘Hoefchen’ and ‘Molino’ soil (each after 0.03 hrs of desorption). Correspondingly the amount of radioactivity in the organic extracts (sum of ambie nt and aggressive extract) increased reaching a ‘s aturation
level’ of about 40 – 45 % of AR for ‘Hoefchen’ soil already after 1 hr of aging and about 48 – 50 % of AR for ‘Molino’ soil at the 5 hrs time point. An extremely fast adsorption process became evident when consid ering the amount of spirotetramat-enol desorbed by the aq ueous solution (mobile fraction of spirotetramat-en ol). Already within the time period of 0.03 hrs necessar y for handling about half of the applied amount of test item was adsorbed to soil and thus only 49.0 and 52.1 % of AR (‘Hoefchen’ and ‘Molino’ soil) were left in t he desorption phase. No spirotetramat-enol was desorba ble with 0.01 M CaCl 2 

### after 5 hrs of aging in both soils.

Due to the very rapid degradation/dissipation of sp irotetramat-enol in the desorption phase in presenc e of soil no stable equilibrium was measurable. Therefor e it was in fact not possible to derive a K d 

### or a K OC 

### using the Freundlich adsorption isotherm. However, using extracted amounts of spirotetramat-enol in soil as adsorbed part and in the limit of detection as the concentration of test item desorbed, the K OC 

### resulted in values of about 1200 L kg -1 

### for ‘Hoefchen’ soil and about 4000 L kg -1 

### for ‘Molino’ soil.

### – 127 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.2.1.2-4: Material balance of radioactivit y [% of AR] in dependence of aging and desorption period (mean value).

Extractable Soil Aging

period [hrs] Desorption

period [hrs] CaCl 2

solution Organic

extract Aggressive

extract Total NER Total
recovery

0.03 52.0 20.6 6.4 79.1 19.8 98.9
1.03 7.9 22.0 16.9 46.8 45.7 92.4
3 5.4 22.0 16.9 44.2 46.8 91.1
5 5.2 19.5 16.7 41.4 52.2 93.6 0

24 5.4 19.9 17.3 42.6 53.7 96.4

0.03 16.1 31.3 8.8 56.2 37.6 93.9 1 24 5.2 30.1 14.7 50.0 44.1 94.1

0.03 6.0 31.4 9.2 46.6 46.5 93.1 5 24 5.4 30.2 13.0 48.6 44.4 93.0

0.03 5.0 29.6 12.0 46.6 46.9 93.5 ‘Hoefchen’

24 24 5.0 30.0 10.7 45.7 49.4 95.1

0.03 55.1 21.3 6.1 82.5 13.7 96.2
1.03 21.8 29.5 14.5 65.7 32.1 97.8
3 8.0 29.0 17.9 54.9 44.1 99.0
5 6.5 29.9 14.6 51.1 47.8 98.9 0

24 4.8 29.6 14.6 49.0 49.6 98.6

0.03 21.9 36.0 9.7 67.6 29.1 96.8 1 24 5.2 35.0 13.2 53.3 45.6 98.9

0.03 8.0 36.4 11.8 56.2 42.0 98.2 5 24 5.2 36.8 12.7 54.7 43.5 98.2

0.03 4.9 36.0 10.9 51.8 44.2 96.0 ‘Molino’

24 24 5.0 34.7 12.5 52.2 43.9 96.1



### Table B.8.2.1.2-5: Characterization of the CaCl 2 

### solution and extractable radioactivity [% of AR] i n dependence of the aging and desorption period (mean value). Spirotetramat-enol Degradates

Soil Aging

period [hrs] Desorption

period [hrs] CaCl 2

solution Both

organic
extracts Total CaCl 2 solution Both

organic
extracts Total

0.03 49.0 14.7 63.7 3.1 12.4 15.5
1.03 2.6 21.2 23.8 5.3 17.7 23.0
3 < LOQ 20.0 20.0 5.4 18.9 24.3

5 < LOQ 18.7 18.7 5.2 17.4 22.6 0

24 < LOQ 18.9 18.9 5.4 18.3 23.7

0.03 11.4 19.3 30.7 4.7 20.8 25.5 1 24 < LOQ 19.3 19.3 5.2 25.5 30.7

0.03 < LOQ 18.6 18.6 6.0 22.0 28.0 5 24 < LOQ 21.4 21.4 5.4 21.7 27.1

0.03 < LOQ 21.7 21.7 5.0 19.9 24.9 ‘Hoefchen’

24 24 < LOQ 17.5 17.5 5.0 23.2 28.2

0.03 52.1 19.3 71.4 3.0 8.1 11.1
1.03 16.8 27.2 44.0 5.0 16.8 21.8
3 < LOQ 27.2 27.2 8.0 19.7 27.7

5 < LOQ 23.5 23.5 6.5 21.1 27.6 0

24 < LOQ 18.7 18.7 4.8 25.5 30.3

0.03 17.9 25.4 43.3 4.0 20.3 24.3 1 24 < LOQ 25.4 25.4 5.2 22.7 27.9

0.03 < LOQ 18.0 18.0 8.0 30.2 38.2 ‘Molino’

5 24 < LOQ 19.9 19.9 5.2 29.5 34.7

### – 128 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Spirotetramat-enol Degradates

Soil Aging

period [hrs] Desorption

period [hrs] CaCl 2

solution Both

organic
extracts Total CaCl 2 solution Both

organic
extracts Total

0.03 < LOQ 15.9 15.9 4.9 30.9 35.8 24 24 < LOQ 18.6 18.6 5.0 28.6 33.6



### Table B.8.2.1.2-6: Calculation of desorbed and adso rbed concentration and of K d 

### and K OC 

### values of spirotetramat-enol in dependence of aging and desor ption period.

Soil Aging

period [hrs] Desorption

period [hrs] C des

[μg mL -1 ] C extr.

[μg g -1 ] K d

[L kg -1 ] K OC

[L kg -1 ]

0.03 0.27 -0.068 a -0.25 -9.7
1.03 0.02 0.061 4.4 169
3 0.002 0.062 32.1 1225
5 0.002 0.058 30.1 1149 0

24 0.002 0.059 30.4 1161

0.03 0.61 0.035 0.6 22 1 24 0.002 0.070 26.9 1386

0.03 0.002 0.058 29.9 1142 5 24 0.002 0.067 34.5 1317

0.03 0.002 0.067 34.9 1332 ‘Hoefchen’

24 24 0.002 0.054 28.0 1069

0.03 0.25 -0.047 a -0.18 -23.1
1.03 0.08 0.051 0.6 79
3 0.011 0.080 7.5 940
5 0.002 0.072 37.3 4659 0

24 0.002 0.058 29.7 3712

0.03 0.097 0.039 0.4 50 1 24 0.002 0.063 32.5 4068

0.03 0.006 0.054 17.7 2208 5 24 0.002 0.061 31.6 3952

0.03 0.002 0.048 16.3 2039 ‘Molino’

24 24 0.002 0.057 29.5 3687

a Theoretically this value should be zero



### Conclusion: Sorption and binding of spirotetramat-enol to soil was extremely fast and increased rapidly with aging time in soil. The portion not tightly bound to the soil, i. e. the portion that is releasable by aqueous soluti on from soil (mobile fraction), is degraded within a few hours ( mainly to spirotetramat-ketohydroxy). From these re sults it can be concluded that spirotetramat-enol is absent from the soil pore water (either degraded or tightl y bound to soil) within a very short period of time.

Reviewer comments:

• Results of this study are considered as additional information, only. Study not considered
essential.

### – 129 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: 14 

### C-BYI08330-ketohydroxy: Adsorption/desorption in fi ve soils Author(s), year: Hein, W., 2005b Study/Report number: M-267955-02-2, IM2001, amended 30.08.2006 Guideline(s): 95/36/EC (1995), OECD Guidelines 106 (2000), US-EPA N 163-1 (1982), Canadian PMRA DACO 8.2.4.2 (1987) 1



### GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: [Azaspirodecenyl-3- 14 

### C]-spirotetramat-ketohydroxy, 4.31 MBq mg -1



### , > 98 % radiochemical purity (HPLC), batch BECH 1588 and 1619 Reference substance: Spirotetramat-ketohydroxy (unl abelled) Soils: Air dried, 2-mm sieved Soil/solution ratio: 1/1, m/v (‘Laacherhof AXXa’, ‘ Laacherhof AIII’) 1/2, m/v (‚Hoefchen’, ‚Molino’, ‚Saskatoon’) Temperature: 20 ± 1 °C Test concentrations: 0.01, 0.03, 0.1, 0.3 and 1.0 m g L -1



### in 0.01 M CaCl 2 

### (containing 50 mg L -1 

### mercuric(II)chloride as biocide)
Co-solvent: None Pre-equilibration time 16 hrs Equilibration time: 24 hrs for adsorption, 24 hrs f or desorption (highest test concentration: 3 desorption cycles) Extraction: After desorption remaining soil was ext racted two times with 20 ml acetonitrile followed by three times with acetonitrile/water (1/ 1, v/v) + 0.5 % formic acid (pH 2.5). After extraction the soil residues were radio -assayed by combustion-LSC. Analytical techniques: LSC, HPLC-UV/RAD, HPLC-MS/MS (ESI) Calculations: The amount adsorbed was calculated by subtracting the equilibrium concentration in the solution from the initial concentration appl ied. The Freundlich adsorption isotherm is described as C s 

### = K F(ads) 

### × C w (1/n) 

### , whereby
C s 

### is the equilibrium concentration of the test item in the soil after adsorption [μg g 1 

### ], C w 

### the equilibrium concentration in the solution [μg mL -1 

### ], K F(ads) 

### is the Freundlich adsorption coefficient and 1/n is the F reundlich exponent. K F(ads) 

### and 1/n were obtained by plotting log(C s 

### ) vs. log(C w 

### ) to give 1/n as the slope and log(K F(ads) 

### ) as the intercept of the linear regression. Desorp tion (based on the desorption constant K F(des) 

### ) was calculated in a similar way. In addition, the linear adsorption coefficient K d(ads) 

### was obtained by linear regression plotting C W 

### vs. C S 

### without data transformation (calculated by the reviewer). Based on the organic carbon [%], the Freundlich ads orption coefficient is expressed as K FOC(ads) 

### = (K F(ads) 

### × 100) / C org 

### (Freundlich desorption and linear adsorption/desorption coefficients are expressed in a similar way).

1 The Canadian guideline is improperly cited in the study report. It should be cited as: Agriculture Can ada (1987) Trade Memorandum T-1-255, Guidelines for Determining Envir onmental Chemistry and Fate of Pesticides.

### – 130 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.2.1.2-7: Physico-chemical characteristics of test soils used for adsorption/desorption of spirotetramat-ketohydroxy.

Soil ‘Laacherhof

AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

Batch ID 20040607 20040607 20040607 81704-S 81004-S

Geographic location Monheim/NorthrhineWestfalia/ Germany Monheim/

NorthrhineWestfalia/ Germany Burscheid/

NorthrhineWestfalia/ Germany Pensacola

Florida/ USA Saskatchewan/

Canada

Texture (USDA) Sandy loam Silt loam Silt loam Sandy loam Clay loam Sand (USDA) [%] 72.7 42.0 12.7 76 28 Silt (USDA) [%] 18.4 44.8 72.5 16 44 Clay (USDA) [%] 8.9 13.2 14.7 8 28 pH (CaCl 2 ) 6.0 6.4 6.1 6.2 5.5 pH (water) 6.6 6.9 6.7 6.3 5.6 pH (KCl) 6.3 6.7 6.4 5.4 4.8 Organic carbon [%] 1.30 1.10 2.62 0.87 2.44 Carbonate [%] < 0.1 < 0.1 < 0.1 0.3 0.7 CEC [meq kg -1 ] 64 96 160 50 196 WHC (0.33 bar) [%] 19.5 23.9 31.3 7.4 23.9 MWHC [%] 38.3 43.5 72.7 23.0 44.5



### Findings: In the pre-test, it was obvious that spirotetramat- ketohydroxy was not stable in the presence of soil. Hence, to prevent microbial degradation of the test item t he test soils were sterilized by soil equilibration in 0.01 M
CaCl 2 

### containing 50 mg L -1 

### mercuric(II)chloride for two days. Under sterile c onditions (preliminary parental mass balance test), spirotetramat-ketohydroxy was s table for 24 hrs of adsorption equilibrium time in 3 soils (test item recovery: 90.7 – 93.7 % of AR). In the ‘ Laacherhof AXXa’ and ‘Laacherhof AIII’ total recove ry of the test item was slightly below 90 % of AR (88.6 % and 87.6 % of AR, respectively). The test item did not
adsorb to the wall surface of the test vessel.

The overall material balance for all concentrations for individual samples was in the range of 92.5 – 98.6 % of AR. After 24 hrs of equilibration 29.8 – 63.0 % of AR was adsorbed to the test soils. The calculate d adsorption constants K F(ads) 

### of the Freundlich isotherms for the five test soil s ranged from 0.516 – 2.21 L kg -1


### (R 2 

### ≥ 0.999), the Freundlich exponent 1/n was in a range of 0.915 – 0.929. The desorption K F(des) 

### values were 1.3 – 1.7 times higher than those obtained for adsorption.

Table B.8.2.1.2-8: Determination of soil/solution r atio : Adsorption of the test item [%] after 24 hrs shaking period at different soil/solution ratios (t est concentration 0.97 mg L -1 

### , mean of replicates).

Soil/solution ratio (m/v) ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

1/1 43.2 39.0 57.7 59.6 75.1
1/2 29.7 25.9 42.6 39.7 58.6

### – 131 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.2.1.2-9: Determination of the adsorption equilibrium time : Adsorption of the test item (expressed as test item equiv. in the adsorption so lution, [mg parent equiv. L -1 

### ]) after a shaking period of 2 – 96 hrs (soil/solution ration 1/1 (m/v ) for ‘Hoefchen’, ‘Molino’ and ‘Saskatoon’ soil and 1/2 (m/v) for ‘Laacherhof AXXa’ and ‘Laacherhof AIII’ s oil, test concentration 0.98 mg L -1 

### , preequilibration time approx. 16 hrs, containing 50 mg L -1 

### mercuric(II)chloride as biocide).

HAT ‘Laacherhof

AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

2 0.65 0.65 0.63 0.72 0.52

4 0.63 0.64 0.62 0.69 0.50
6 0.63 0.64 0.62 0.68 0.48
24 0.59 (0.54) a 0.60 (0.56) 0.59 (0.54) 0.63 (0.59) 0.44 (0.42)
30 0.59 0.61 0.58 0.64 0.42
48 0.58 (0.54) 0.60 (0.55) 0.58 (0.53) 0.60 (0.56) 0.4 1 (0.39)
72 0.56 (0.52) 0.59 (0.54) 0.57 (0.53) 0.59 (0.55) 0.4 0 (0.37)
96 0.57 (0.53) 0.59 (0.54) 0.57 (0.53) 0.59 (0.56) 0.3 8 (0.34)

a Number in parentheses give test item concentration [mg parent L -1 ]



### Table B.8.2.1.2-10: Determination of the desorption equilibrium time : Desorption of the test item (expressed as test item equiv. in the adsorption so lution, [mg parent equiv. L -1 

### ]) after a shaking period of 2 – 72 hrs (soil/solution ration 1/1 (m/v ) for ‘Hoefchen’, ‘Molino’ and ‘Saskatoon’ soil and 1/2 (m/v) for ‘Laacherhof AXXa’ and ‘Laacherhof AIII’ s oil, test concentration 0.93 mg L -1 

### , preequilibration time approx. 16 hrs, adsorption time 24 hrs, containing 50 mg L -1 

### mercuric(II)chloride as biocide).

HAT ‘Laacherhof

AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

2 0.29 0.28 0.27 0.19 0.21

4 0.29 0.28 0.27 0.20 0.22
6 0.29 0.30 0.22 0.25 0.22
24 0.30 0.30 0.28 0.20 0.22
48 0.29 0.30 0.27 0.21 0.22
72 0.28 0.29 0.27 0.20 0.21



### Table B.8.2.1.2-11: Parental mass balance : Recover of test item [% of AR] in soil after the preliminary adsorption equilibrium test and extraction (24 hrs equilibrium time, soil/solution ration 1/1 (m/v) fo r ‘Hoefchen’, ‘Molino’ and ‘Saskatoon’ soil and 1/2 ( m/v) for ‘Laacherhof AXXa’ and ‘Laacherhof AIII’ soil, test concentration 0.98 mg L -1 

### , pre-equilibration time approx. 16 hrs, containing 50 mg L -1 

### mercuric(II)chloride as biocide).

Compartment ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

Solution 37.6 37.1 42.2 51.0 33.9
Extraction 1 45.9 44.2 44.5 36.5 53.4
Extraction 2 5.1 6.3 4.0 3.6 6.3 Test item

Recovery 88.6 87.6 90.7 91.2 93.7

Unidentified RA in solution 3.5 3.1 3.6 3.2 1.6
Unidentified RA in extracts 0.0 0.2 0.0 0.0 0.0

NER 1.1 1.6 1.1 0.9 1.7 Total recovery 93.2 92.5 95.3 95.2 96.9– 132 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.2.1.2-12: Calculated non-linear (Freundlich) and linear adsorptionisotherm parameter for spirotetramat-ketohydroxy.Non-linear isotherm (Freundlich) Linear isothermSoil Texture pH (CaCl2) pH (sup.)aCorg[%]KF(ads)[L kg-1]KFOC(ads)[L kg-1]1/n R2Kd(ads)[L kg-1]KOC(ads)[L kg-1]R2‘Laacherhof AXXa’ Sandy loam 6.0 6.7 1.30 0.533 41.0 0.920 1.000 0.560 43.1 0.998‘Laacherhof AIII’ Silt loam 6.4 6.9 1.10 0.516 46.90.929 1.000 0.529 48.1 0.997‘Hoefchen’ Silt loam 6.1 6.6 2.62 1.08 41.2 0.927 1.000 1.10 42.0 0.996‘Molino’ Sandy loam 6.2 6.7 0.87 0.862 99.1 0.918 1.000 0.867 99.7 0.994‘Saskatoon’ Clay loam 5.5 6.0 2.44 2.21 90.4 0.9151.000 2.28 93.4 0.994apH in the supernatant of the test solution.Table B.8.2.1.2-13: Calculated non-linear (Freundlich) desorptionisotherm constants for spirotetramat-ketohydroxy.1stDesorptionSerial desorption(only highest test concentration)Soil TexturepH(CaCl2)pH (sup.)aCorg[%]KF(des)[L kg-1]KFOC(des)[L kg-1]1/n R2KFser[L kg-1]KFOCser[L kg-1]1/n R2‘Laacherhof AXXa’ Sandy loam 6.0 6.7 1.30 0.668 51.4 0.933 1.000 0.442 34.0 0.566 0.999‘Laacherhof AIII’ Silt loam 6.4 6.9 1.10 0.713 64.80.954 0.999 0.415 37.7 0.522 1.000‘Hoefchen’ Silt loam 6.1 6.6 2.62 1.60 61.2 0.983 1.000 0.922 35.2 0.581 0.992‘Molino’ Sandy loam 6.2 6.7 0.87 1.47 170 0.885 0.999 0.636 73.1 0.312 0.998‘Saskatoon’ Clay loam 5.5 6.0 2.44 2.84 116 0.902 0.997 1.51 61.7 0.509 0.999apH in the supernatant of the test solution.

### – 133 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion: The adsorption constants K F(ads) 

### of spirotetramat-ketohydroxy calculated based on t he Freundlich isotherms for the five test soils ranged from 0.516 – 2.21 L kg -1 

### . The respective calculated K FOC(ads) 

### values were in the range of 41.0 – 99.1 L kg -1 

### , 1/n values were in a range of 0.915 – 0.929. Spir otetramat-ketohydroxy is
classified as highly mobile according to the classification scheme of McCall e t al. (1981) 1 

### and mobile according to FAO 2 

### classification.

Reviewer comments:

• Linear adsorption constants (K OC(ads) 

### ) were close to the Freundlich adsorption constants with a range of 42.0 – 99.7 L kg -1



### .

Reference: [ 14 

### C]-BYI08330-MA-amide: Adsorption to and desorption from five soils Author(s), year: Simmonds, M., Early, E., 2005 Study/Report number: M-263686-01-2, CX/04/070 Guideline(s): OECD Guidelines 106 (2000), US-EPA N 163-1 (1982), Canadian PMRA DACO 8.2.4.2 (1987) 3 

### , Japanese MAFF new Test Guidelines for Supporting Registration of Pesticides (2000 and 2001)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: [ 14 

### C]-spirotetramat-MA-amide (see structure for labell ing, which is indicated by an asterisk), 4.08 MBq mg -1 

### , > 98 % radiochemical purity (HPLC), batch BECH 16 21

Reference substance: Spirotetramat-MA-amide (unlabe lled) Soils: Air dried, 2-mm sieved Soil/solution ratio: 1/1, m/v Temperature: 24 ± 2 °C (adsorption), 20 ± 2 °C (des orption) Test concentrations: 0.005, 0.01, 0.05, 0.1 and 0.5 mg L -1



### in 0.01 M CaCl 2 

### Co-solvent: Acetonitrile ( ≤ 0.4 %, v/v) Pre-equilibration time 16 hrs Equilibration time: 24 hrs for adsorption, 2 hrs fo r desorption (highest test concentration: 3 desorpt ion cycles) Extraction: After desorption remaining soil was ext racted twice with 40 ml of acetonitrile/water (80/20, v/v, adjusted to pH 4 with formic acid) fol lowed by one extraction with 40

1 McCall, J.P., D.A. Laskowsky, R.L. Swann, and H.J. D ishburger. 1981. Measurement of sorption coefficien ts of organic chemicals and their use in environmental fate analy sis. Pages 89-109 In Test protocols for environmental fate & movement of toxicants. Proceedings of a symposium. Association of Official Analytical Chemists. 94 th Annual Meeting, October 21-22, 1980. Washington, DC.
2 Food and Agriculture Organization of the United Na tions. FAO PESTICIDE DISPOSAL SERIES 8. Assessing Soil Contamination: A Reference Manual. Appendix 2. Paramet ers of pesticides that influence processes in the s oil. Editorial Group, FAO Information Division: Rome, 2000.
3 The Canadian guideline is improperly cited in the study report. It should be cited as: Agriculture Can ada (1987) Trade Memorandum T-1-255, Guidelines for Determining Envir onmental Chemistry and Fate of Pesticides.

### – 134 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

ml of acetonitrile. After extraction the soil resid ues were radio-assayed by
combustion-LSC. Analytical techniques: LSC, HPLC-UV/RAD, LC-MS/MS
LOQ = 0.004 % of AR Calculations: The amount adsorbed was calculated by subtracting the equilibrium concentration in the solution from the initial concentration appl ied. The Freundlich adsorption isotherm is described as C s 

### = K F(ads) 

### × C w (1/n) 

### , whereby
C s 

### is the equilibrium concentration of the test item in the soil after adsorption [μg g -1 

### ], C w 

### the equilibrium concentration in the solution [μg mL -1 

### ], K F(ads) 

### is the Freundlich adsorption coefficient and 1/n is the Fr eundlich exponent. K F(ads) 

### and 1/n were obtained by plotting log(C s 

### ) vs. log(C w 

### ) to give 1/n as the slope and log(K F(ads) 

### ) as the intercept of the linear regression. Desorp tion (based on the desorption constant K F(des) 

### ) was calculated in a similar way. In addition, the linear adsorption coefficient K d(ads) 

### was obtained by linear regression plotting C W 

### vs. C S 

### without data transformation (calculated by the reviewer). Based on the organic carbon [%], the Freundlich ads orption coefficient is expressed as K FOC(ads) 

### = (K F(ads) 

### × 100) / C org 

### (Freundlich desorption and linear adsorption/desorption coefficients are expressed in a similar way).

Table B.8.2.1.2-14: Physico-chemical characteristic s of test soils used for adsorption/desorption of spirotetramat-MA-amide.

Soil ‘Laacherhof

AXXa’ ‘Laacherhof

AIIIa’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

Batch ID 04/73 04/72 04/71 04/67 04/68

Geographic location Monheim/NorthrhineWestfalia/ Germany Monheim/

NorthrhineWestfalia/ Germany Burscheid/

NorthrhineWestfalia/ Germany Pensacola

Florida/ USA Saskatchewan/

Canada

Texture (USDA) Sandy loam Silt loam Silt loam Loamy sand Loam
Soil taxonomic classification (USDA) Sandy, mixed, mesic Typic Cambudoll Loamy, mixed,

mesic Typic Argudalfs Loamy, mixed,

mesic Typic Argudalfs Ultisol na

Sand (USDA) [%] 72.7 45.0 12.7 78 30 Silt (USDA) [%] 18.4 40.1 72.5 15 45 Clay (USDA) [%] 8.9 14.9 14.7 7 25 pH (CaCl 2 ) 6.3 6.7 6.5 5.7 5.4 pH (water) 6.9 7.5 7.1 6.1 5.7 pH (KCl) 6.3 6.7 6.3 Nd nd Organic carbon [%] 1.70 0.91 2.07 0.7 2.3 CEC [meq kg -1 ] 89 87 141 47 212 WHC (0.33 bar) [%] 19.5 23.9 31.3 9.0 24.7 MWHC [%] 42.3 38.7 66.2 nd nd



### Findings: No significant breakdown of spirotetramat-MA-amide in any soil was observed (> 90 % as test-item).

The overall material balance for all concentrations for individual samples was in the range of 90.5 – 100.9 % of AR. After 24 hrs of equilibration 4.7 – 21.6 % o f AR was adsorbed to the test soils. The calculated adsorption constants K F(ads) 

### of the Freundlich isotherms for the five test soil s ranged from 0.06 – 0.18 L kg -1


### (R 2 

### ≥ 0.998), the Freundlich exponent 1/n was in a range of 0.80 – 1.07. The desorption K F(des) 

### values were 1.6 – 2.3 times higher than those obtained for adso rption.

### – 135 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.2.1.2-15: Determination of the soil/solut ion ratio : Adsorption of the test item [%] after 24 hrs shaking period at different soil/solution ratios (t est concentration 0.50 mg L -1 

### ).

Soil/solution ratio (m/v) ‘Laacherhof AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

1/1 4.8 4.4 9.2 14.4 10.7
1/2 3.0 5.7 6.0 9.5 6.0
1/3 1.8 6.8 3.4 7.0 4.9



### Table B.8.2.1.2-16: Determination of the adsorption equilibrium time : Adsorption of the test item [%] after a shaking period of 24 – 144 hrs (soil/soluti on ration 1/1 (m/v), test concentration 0.50 mg L -1



### , pre-equilibration time approx. 16 hrs).

HAT ‘Laacherhof

AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

24 6.7 6.2 11.2 14.3 8.2
48 6.2 6.9 8.7 14.8 10.0
72 7.1 8.1 8.8 17.4 13.9
96 7.8 7.4 8.7 20.0 16.5
120 8.5 6.8 8.5 21.0 17.1

144 7.3 7.8 9.4 22.4 16.6



### Table B.8.2.1.2-17: Determination of the desorption equilibrium time : Desorption [%] of the test item after a shaking period of 1 – 48 hrs (soil/solution ration 1/1 (m/v), test concentration 0.50 mg L -1 

### , preequilibration time approx. 16 hrs, adsorption time 48 hrs).

HAT ‘Laacherhof

AXXa’ ‘Laacherhof

AIII’ ‘Hoefchen’ ‘Molino’ ‘Saskatoon’

1 31.2 29.4 28.2 28.6 28.6
2 32.0 30.6 32.2 29.5 36.1
24 31.2 29.3 32.2 29.7 28.5
48 30.7 29.9 33.1 29.5 34.9– 136 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.2.1.2-18: Calculated non-linear (Freundlich) and linear adsorptionisotherm parameter for spirotetramat-MA-amide.Non-linear isotherm (Freundlich) Linear isothermSoil Texture pH (CaCl2) pH (sup.)aCorg[%]KF(ads)[L kg-1]KFOC(ads)[L kg-1]1/n R2Kd(ads)[L kg-1]KOC(ads)[L kg-1]R2‘Laacherhof AXXa’ Sandy loam 6.3 nd 1.7 0.075 4.4 1.07 1.000 0.071 4.2 0.997‘Laacherhof AIII’ Silt loam 6.7 nd 0.91 0.059 6.5 0.96 0.999 0.064 7.0 0.998‘Hoefchen’ Silt loam 6.5 nd 2.07 0.104 5.0 0.80 0.998 0.116 5.6 0.976‘Molino’ Loamy sand 5.7 nd 0.7 0.179 25.5 0.98 0.999 0.179 25.6 0.997‘Saskatoon’ Loam 5.4 nd 2.3 0.117 5.1 0.93 0.999 0.126 5.5 0.994apH in the supernatant of the test solution.Table B.8.2.1.2-19: Calculated desorptionisotherm parameter for spirotetramat-MA-amide.1stDesorptionSerial desorption(only highest test concentration)bSoil TexturepH(CaCl2)pH(sup.)aCorg[%]KF(des)[L kg-1]KFOC(des)[L kg-1]1/n R2KFser[L kg-1]KFOCser[L kg-1]1/n‘Laacherhof AXXa’ Sandy loam 6.3 nd 1.7 0.13 7.8 0.99 0.994 0.04 2.4 0.229‘Laacherhof AIIIa’ Silt loam 6.7 nd 0.91 0.14 14.90.96 0.994 0.04 4.4 0.183‘Hoefchen’ Silt loam 6.5 nd 2.07 0.19 9.0 0.80 1.000 0.06 2.9 0.165‘Molino’ Loamy sand 5.7 nd 0.7 0.37 52.6 0.98 0.9980.10 14.3 0.255‘Saskatoon’ Loam 5.4 nd 2.3 0.22 9.7 0.91 0.999 0.07 3.0 0.180apH in the supernatant of the test solutionbCalculated by the reviewer

### – 137 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion: The adsorption constants K F(ads) 

### of spirotetramat-MA-amide calculated based on the Freundlich isotherms for the five test soils ranged from 0.06 – 0.18 L kg -1 

### . The respective calculated K FOC(ads) 

### values were in the range of 4.4 – 25.5 L kg -1 

### , 1/n values were in a range of 0.80 – 1.07. Spirot etramat-MA-amide is classified as very highly mobile

according to the classification scheme of McCall e t al. (1981) 1 

### and highly mobile according to
FAO 2 

### classification.

Reviewer comments:

• Linear adsorption constants (K OC(ads) 

### ) were close to the Freundlich adsorption constants with a range of 4.2 – 25.6 L kg -1



### .

Reference: Estimation of the adsorption coefficient (KOC) of B YI 08330-enol dimer 1 (ROI 6, 14 

### C-labelled) and BYI 08330-enol dimer 2 (ROI 7, 14 

### C-labelled) on soil using High performance Liquid Chromatography (HPLC) Author(s), year: Neumann, B., Klempner, A., 2006 Study/report number: M-274184-01-2, MEF-06/123, ame nded 01.06.2007 Guideline(s): OECD Guideline 121 (2001)
GLP: Yes
Deviations: None Validity: Study considered as additional information, only – study not essential

Material and methods:
Test substances: • BYI 08330-enol dimer 1, 0.74 MBq mg -1



### • BYI 08330-enol dimer 2, 0.74 MBq mg -1 

### Both test items isolated from aerobic soil degradat ion/metabolism study conducted with spirotetramat in four soils (Babczin ski, 2005)
Reference substances: NaNO 3 

### , aldicarb, atrazine, spirotetramat-ketohydroxy, sp irotetramat, methiocarb, triadimenol, linuron, spirodiclofen-ketohydroxy, py razophos, phenanthrene, spirodiclofen, 4,4’-DDT Chromatography
column: ZORBAX CN, 5 μm, 250 × 4.6 mm

Mobile phase: • pH 6.0: methanol/0.01 M citrate buffer (55/45, v/v )

• pH 1.7: methanol/0.05 M phosphoric acid buffer (55 /45, v/v) Isocratic elution, 1.5 mL min -1 

### Calculation: Capacity factor (k’) = (RT substance 

### – RT NaNO3 

### )/ RT NaNO3 

### , RT NaNO3 

### is regarded as column dead time, regression analysis log(k’) vs. l og(K OC 

### )

1 McCall, J.P., D.A. Laskowsky, R.L. Swann, and H.J. D ishburger. 1981. Measurement of sorption coefficien ts of organic chemicals and their use in environmental fate analy sis. Pages 89-109 In Test protocols for environmental fate & movement of toxicants. Proceedings of a symposium. Association of Official Analytical Chemists. 94 th Annual Meeting, October 21-22, 1980. Washington, DC.
2 Food and Agriculture Organization of the United Na tions. FAO PESTICIDE DISPOSAL SERIES 8. Assessing Soil Contamination: A Reference Manual. Appendix 2. Paramet ers of pesticides that influence processes in the s oil. Editorial Group, FAO Information Division: Rome, 2000.

### – 138 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Findings: Average capacity factors (k’) were derived for each compound and a linear function was established for log(k’) values vs. log(K OC 

### ) values with a slope of 0.253 and 0.295 at pH 6.0 and pH 1.7, respectively, and an intercept of -0.478 and -0.628, respectively (R 2 

### ≥ 0.935).

y = 0.2528 x - 0.478

R 2 = 0.9348

-0.2 0.0 0.2 0.4 0.6 0.8 1.0

0 1 2 3 4 5 6

log(K OC )log(k') Reference substances Spirotetramat-enol dimer 1 Spirotetramat-enol dimer 2

y = 0.2948 x - 0.6284

R 2 = 0.9665

-0.2 0.0 0.2 0.4 0.6 0.8 1.0

0 1 2 3 4 5 6

log(K OC )log(k') Reference substances Spirotetramat-enol dimer 1 Spirotetramat-enol dimer 2



### Figure B.8.2.1.2-1: Calibration plots log(k’) vs. l og(K OC 

### ) and linear regression lines (left: at pH 6.0, right: at pH 1.7).

Table B.8.2.1.2-20: Calculated K OC 

### values [L kg -1 

### ] of spirotetramat-enol dimer 1 and dimer 2 accordi ng to the HPLC method (corrected by the reviewer). Spirotetramat-enol dimer 1 Spirotetramat-enol dimer 2 System log(k’) log(K OC ) K OC [L kg -1 ] log(k’) log(K OC ) K OC [L kg -1 ] pH 6.0 0.343 3.25 1771 0.405 3.49 3115 pH 1.7 0.306 3.17 1477 0.409 3.52 3301

Arithmetic mean - - 1624 - - 3208



### Conclusion: According to the HPLC method, spirotetramat-enol di mer 1 and spirotetramat-enol dimer 2 can be
considered as immobile in soil with estimated K OC 

### values of approx. 1624 L kg -1 

### and 3208 L kg -1 

### . It can be concluded that both test items do not have ionic (a cidic or alkaline) properties. Spirotetramat-enol d imer 1 is
classified as lowly mobile according to the classification scheme of McCall e t al. (1981) and slightly mobile according to FAO classification, spirotetramat-enol dimer 2 is classified as slightly mobile according to the classification scheme of McCall et al. (1981) 1 

### and according to FAO 2 

### classification.

Reviewer comments:

• Study not considered essential. Study results included for additional information o nly.

• In the study report, log(K OC 

### ) was incorrectly set as the dependent variable (y axis) in the calibration plots/curves. This was corrected by the reviewer (r esulting in a slight change of the results).

• Reverence substances also include spirotetramat an d spirotetramat-ketohydroxy which were both
well in line with the other test substances confirm ing the results from individual adsorption/desorpti on batch equilibrium experiments with these two substa nces.

1 McCall, J.P., D.A. Laskowsky, R.L. Swann, and H.J. D ishburger. 1981. Measurement of sorption coefficien ts of organic chemicals and their use in environmental fate analy sis. Pages 89-109 In Test protocols for environmental fate & movement of toxicants. Proceedings of a symposium. Association of Official Analytical Chemists. 94 th Annual Meeting, October 21-22, 1980. Washington, DC.
2 Food and Agriculture Organization of the United Na tions. FAO PESTICIDE DISPOSAL SERIES 8. Assessing Soil Contamination: A Reference Manual. Appendix 2. Paramet ers of pesticides that influence processes in the s oil. Editorial Group, FAO Information Division: Rome, 2000.

### – 139 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.2.1.3 Column leaching studies with the active s ubstance (OECD Annex IIA, 7.4.3, IIIA, 9.3.1)

Column leaching studies were not performed for the active studies. Reliable adsorption/desorption para meter could be derived with an equilibrium batch study (B .8.2.1.1).

B.8.2.1.4 Column leaching studies with relevant met abolites (OECD Annex IIA, 7.4.4, IIIA, 9.3.1)

A column leaching studies was performed with spirot etramat-enol on four soils using [3- 14 

### C] labelled test item. This study was investigated in an additional modelling study using the transport model HYDRUS.

Reference: BYI 08330-enol: Soil column leaching Author(s), year: Babczinski, P., 2006c Study/report number: M-270280-02-2, MEF-05/356, ame nded 22.05.2006 Guideline(s): OECD Guideline 312 (2003), US-EPA N § 163-1 (1982), SETAC (1995), CAN Environmental Chemistry and Fate – Guidelines for R egistration of Pesticides (1987), PMRA Harmonization of environmental chemist ry and fate data requirements under NAFTA (2001)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: [Azaspirodecenyl-3- 14 

### C]-spirotetramat-enol, 4.54 MBq mg -1



### , > 99 % of AR radiochemical purity (HPLC), batch BEC H 1610 Reference substances: Spirotetramat-enol (unlabelle d), spirotetramat-ketohydroxy, spirotetramat-MAamide Application rate: 30.3 μg spirotetramat-enol column -1 

### (equivalent to 153 g met ha -1 

### under the assumption of 288 g ai ha -1 

### , a worst case formation rate of 50 % and additiona l 1/3 amount owing to analytical reasons) Soil column: 5 cm diameter, 30 cm length, soil air dried soil: 583 – 836 g corresponding to 760 – 900 g fresh soil, column leaching tests were perf ormed in duplicate Column conditions: Saturated flow conditions, level of ponding CaCl 2 

### solution approx. 1 cm
Elution: 1000 ml of 0.01 M CaCl 2 

### solution (corresponding to 50.8 cm, pH approx. 6) over a period of 5 days (equivalent to approx. 200 ml day -1 

### ), leachate collected in 20 ml fractions, each group of 5 fractions combined to yi eld 10 × 100 ml fractions, residual eluent from the column (up to 130 cm) addi tionally collected (11 th 

### fraction) Column handling: After termination of the leaching process, the column was frozen and cut into 5 × 6 cm segments. Soil segments were extracted three t imes with 120 ml of acetonitrile/water (1/1, v/v) containing 0.1 % ammo nium chloride (w/v) and 0.06 % ammonia (v/v), pH approx. 8.6. The extracts were co mbined and radio-assayed by
LSC. The soil residues were further extracted with 120 ml of the same extractant using a microwave extractor for 10 min at 70 °C, th e extract was radio-assayed by LSC. All extracts were concentrated (SpeedVac) and analysed by HPLC. Remaining soil debris was radio-assed by combustion -LSC. Volatile traps: Not in the scope of the study, not applied Analytical techniques: LSC, HPLC-UV/RAD, LC-MS/MS
LOQ ≤ 0.24 % of AR (HPLC, soil segments), ≤ 0.02 % (HPLC, leachates)
Calculations: K d 

### values of spirotetramat-enol (mobile fraction) wer e calculated using the following formula (Ketelle and Boyd, 1947 [1]; Swob oda and Thomas, 1968 [2]):

### – 140 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008



### W V

V V k v

v p d 

### ⋅

       

− = 1

V p 

### = Volume of water required to leach 50 % of the te st compound through the soil column [mL]
V v 

### = Void volume of the column [mL] W = Soil dry weight in the column [g]

[1] Ketelle, B. H., Boyd, G. E. (1947) The exchange adsorption of ions from aqueous solutions by organic zeolithes. IV. The sep aration of the Yttrium group
rare earths. J. Amer. Chem. Soc. 69 , 2800-2812 [2] Swoboda, A. R., Thomas, G. W. (1968) Movement o f parathion in soil columns. J. Agric. Food Chem. 16 , 923-927

Table B.8.2.1.4-1: Physico-chemical characteristics of test soils used for the column study with spirotetramat-enol.

Soil ‘Laacherhof AXXa’ ‘Laacherhof AIIIa’ ‘Hoefchen ’ ‘Molino’

Batch ID 20040929 20040929 20040424 20040427

Geographic location Monheim/ Northrhine-Westfalia/ Germany Monheim/

Northrhine-Westfalia/ Germany Burscheid/

Northrhine-Westfalia/ Germany Pensacola Florida/ USA

Texture (USDA) Sandy loam Loam Silt loam Sandy loam
Soil taxonomic classification (USDA) Sandy, mixed, mesic Typic Cambudolls Sandy, mixed, mesic Typic Cambudolls Loamy, mixed, mesic Typic Argudalfs Fine-loamy, kaolinitic, thermic Plinthic Kandiudults SCS Soil series Mollisols, Udolls Alfisols, Udalfs Alfis ols, Udalfs Ultisol Sand (USDA) [%] 69 49 19 77 Silt (USDA) [%] 21 31 63 13 Clay (USDA) [%] 10 20 18 10 pH (CaCl 2 ) 6.1 6.5 6.4 5.4 pH (water) 6.6 7.1 6.8 6.0 pH (KCl) 6.1 6.4 6.2 5.4 pH (satd. paste) 6.6 7.1 6.8 na Organic carbon [%] 2.0 1.2 2.4 0.8 CEC [meq kg -1 ] 109 102 141 42 WHC (0.33 bar) [%] 13.5 17.1 25.6 8.8 MWHC [%] 29.1 25.7 67.5 22.6



### Findings: The total recovery of radioactivity of the column i ncluding the leachates was in a range of 86.9 – 96. 8 % of AR (individual values). Since no volatile traps wer e used complete recovery is not achievable. The tot al radioactivity detected in the leachates was in a ra nge of 2.6 – 17.0 % of AR; from the column segments 70.0
– 93.4 % of AR could be recovered. Total NER in col umn segments was in a range of 29.5 – 42.2 % of AR.

Table B.8.2.1.4-2: Material balance [% of AR] at th e end of the column study (5 DAT, mean of two replicates).

Soil column

Soil Extractable NER Sub-total Leachate Total

‘Laacherhof AXXa’ 37.8 42.2 80.1 12.3 92.4
‘Laacherhof AIIIa’ 33.5 36.5 70.0 17.0 87.0
‘Hoefchen’ 41.3 40.0 81.3 8.9 90.2
‘Molino’ 63.8 29.5 93.4 2.6 96.0

### – 141 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

In the soil column recovered radioactivity was focu ssed on the top soil segment (0 – 6 cm), 64.2 – 77. 6 % of AR was found within the uppermost segment. The rema ining 5 segments (6 – 30 cm) covered 5.8 – 15.8 %
of AR.

Table B.8.2.1.4-3: Characterization of total and ex tractable radioactivity [% of AR] in soil column segments at the end of the column study (5 DAT, mean of two replicates). Soil column segment [cm] Soil 0 – 6 6 – 12 12 – 18 18 – 24 24 – 30 Total

‘Laacherhof AXXa’ 69.4 2.5 2.3 2.8 3.1 80.1
‘Laacherhof AIIIa’ 64.2 2.3 1.1 1.1 1.3 70.0
‘Hoefchen’ 70.9 3.3 2.0 2.2 2.8 81.3 Total radioactivity

‘Molino’ 77.6 9.2 3.9 1.8 0.8 93.4

‘Laacherhof AXXa’ 29.3 1.6 1.8 2.4 2.6 37.8
‘Laacherhof AIIIa’ 29.6 1.4 0.7 0.8 1.0 33.5
‘Hoefchen’ 33.6 2.3 1.4 1.7 2.3 41.3 Extractable radioactivity

‘Molino’ 49.7 8.4 3.5 1.6 0.6 63.8

‘Laacherhof AXXa’ 10.2 nd nd nd nd 10.2
‘Laacherhof AIIIa’ 9.6 nd nd nd nd 9.6
‘Hoefchen’ 11.5 0.1 nd nd nd 11.6Spirotetramatenol

‘Molino’ 16.5 1.4 nd nd nd 17.9

‘Laacherhof AXXa’ 1.4 1.1 1.5 2.0 2.4 8.4
‘Laacherhof AIIIa’ 0.8 0.7 0.5 0.6 0.7 3.3
‘Hoefchen’ 1.8 0.8 1.0 1.3 1.8 6.7Spirotetramatketohydroxy

‘Molino’ 3.7 2.9 2.8 1.1 0.5 11.0

‘Laacherhof AXXa’ nd nd 0.1 0.1 0.1 0.3
‘Laacherhof AIIIa’ nd nd nd nd nd nd
‘Hoefchen’ 0.1 0.1 0.1 0.2 0.1 0.6SpirotetramatMA-amide

‘Molino’ nd nd 0.1 0.1 nd 0.2

‘Laacherhof AXXa’ 8.8 nd nd nd nd 8.8
‘Laacherhof AIIIa’ 9.3 0.3 nd nd nd 9.6
‘Hoefchen’ 9.4 0.2 nd nd nd 9.6Spirotetramatenol-dimer 1

‘Molino’ 11.8 1.9 nd nd nd 13.7

‘Laacherhof AXXa’ 4.8 nd nd nd nd 4.8
‘Laacherhof AIIIa’ 5.9 nd nd nd nd 5.9
‘Hoefchen’ 6.1 nd nd nd nd 6.1Spirotetramatenol-dimer 2

‘Molino’ 10.7 0.1 nd nd nd 10.8

‘Laacherhof AXXa’ 4.1 0.6 0.2 0.3 0.4 5.6
‘Laacherhof AIIIa’ 3.9 0.4 0.2 0.2 0.3 5.0
‘Hoefchen’ 4.5 1.0 0.3 0.2 0.5 6.5 Unidentified radioactivity

‘Molino’ 7.0 2.1 0.5 0.4 0.1 10.1– 142 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.2.1.4-4: Amounts of total radioactivity,spirotetramat-enol, spirotetramat-ketohydroxy and spirotetramat-MA-amide in leachate fractions[% of AR] (5 DAT, mean of two replicates).Leachate fractionaSoil1 2 3 4 5 6 7 8 9 10 11 Total‘Laacherhof AXXa’ < 0.1 < 0.1 0.3 1.51.81.5 1.3 1.2 1.4 1.6 1.6 12.3‘Laacherhof AIIIa’ < 0.1 0.1 0.9 1.9 2.62.92.7 2.2 1.7 1.3 0.6 17.0‘Hoefchen’ < 0.1 < 0.1 < 0.1 0.3 0.7 1.0 1.1 1.1 1.3 1.51.98.9Totalradioactivity‘Molino’ < 0.1 < 0.1 0.1 0.40.50.4 0.3 0.3 0.2 0.2 0.2 2.6‘Laacherhof AXXa’ na na na 0.1 0.30.5 0.5 0.50.4 0.3 0.3 2.8‘Laacherhof AIIIa’ na na < 0.1 nd < 0.1 < 0.1 0.20.3 0.3 0.30.2 1.3‘Hoefchen’ na na na nd 0.1 0.3 0.4 0.40.50.40.52.7Spirotetramatenol‘Molino’ na na na nd nd nd nd < 0.1 < 0.1 < 0.1 < 0.1 0.1‘Laacherhof AXXa’ na na na < 0.1 0.1 < 0.1 0.1 0.30.6 0.81.02.9‘Laacherhof AIIIa’ na na < 0.1 0.3 1.01.6 1.61.2 0.9 0.7 0.3 7.5‘Hoefchen’ na na na nd < 0.1 < 0.1 < 0.1 0.1 0.3 0.50.92.0Spirotetramatketohydroxy‘Molino’ na na na nd < 0.1 < 0.1 < 0.1 < 0.1 < 0.1< 0.1 < 0.1 0.2‘Laacherhof AXXa’ na na na0.80.4 0.2 0.1 0.1 0.1 0.1 0.1 1.9‘Laacherhof AIIIa’ na na0.2 0.2nd < 0.1 0.1 0.1 0.1 < 0.1 < 0.1 0.8‘Hoefchen’ na na na 0.10.2 0.2 0.20.1 0.1 0.1 0.1 1.0SpirotetramatMA-amide‘Molino’ na na na 0.30.40.3 0.2 0.1 0.1 0.1 0.1 1.5aEach leachate fraction corresponds to 100 mL

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.2.1.4-5: Calculated K OC 

### [L kg -1 

### ] values for spirotetramat-enol from the column exp eriments.

Soil Repl. V p [mL] V v [mL] W [g] K d [L kg -1 ] K OC [L kg -1 ] Mean

A 675 327 669 0.52 26 ‘Laacherhof AXXa’ B 715 340 669 0.56 28 27

A 858 304 710 0.78 65 ‘Laacherhof AIIIa’ B 864 312 710 0.78 65 65

A 849 368 583 0.83 34 ‘Hoefchen’ B 701 368 583 0.57 24 29

A 861 260 836 0.72 90 ‘Molino’ B 994 272 836 0.86 108 99

V p = Volume of water required to leach 50 % of the te st compound through the soil column (including void volume) [mL] V v = Void volume of the column [mL] W = Soil dry weight in the column [g]



### The test item spirotetramat-enol was almost exclusi vely located in the top soil segment (9.6 – 16.5 % of AR). Only in the ‘Molino’ soil a small amount of spirote tramat-enol (1.4 % of AR) was detected in the secon d soil segment. In the other three soils spirotetramat-eno l was not detected in deeper column segments ( ≤ 0.1 % of AR).

Other major components detected in the top segment extracts were spirotetramat-enol dimer 1 (8.8 – 11. 8 % of AR) and spirotetramat-enol dimer 2 (4.8 – 10. 7 % of AR) for which strong evidence is given that th eir formation is artifactual owing to the application t echnique. Spirotetramat-enol dimer 1 exhibited a li ttle potential to move into the second soil segment at v ery low extent, spirotetramat-enol dimer 2 was comp letely
immobile.

Spirotetramat-ketohydroxy was recovered from the to p soil segments in a range of 0.8 – 3.7 % of AR. In contrast to the spirotetramat-enol and the dimers o f spirotetramat-enol, spirotetramat-ketohydroxy was present in the further segments of all soils after leaching, indicating a certain translocation potent ial. In ‘Molino’ soil a decreasing gradient towards the col umn end was observed, whereas in soils ‘Laacherhof AXXa’ and ‘Hoefchen’ this gradient increased. Its b ehaviour in ‘Laacherhof AIIIa’ soil was less clear.

Spirotetramat-MA-amid was formed from spirotetramat -enol in low amounts. In three of the four soils it was found at maximum 0.2 % of AR. This compound had a t endency to move down the columns as well and was hardly detected in any of the top and second soil s egments.

Conclusion: From this study, it is evident that only a tiny por tion of spirotetramat-enol had a potential to leach into deeper soil layers or leave the soil column, whereas by fa r the major portion strongly bound to the top soil layer. Only a minor part of the applied spirotetramat-enol was recovered in soil extracts and even smaller po rtion in the leachate, indicating immediate and strong bindi ng (chemisorption) to the top soil and to a lower e xtent transformation into spirotetramat-enol derived degr adates, but a significant formation of NER during t he progress of the study. For the mobile (non-bound) s pirotetramat-enol fraction K OC 

### values between 27 – 99 L kg -1 

### were calculated. Spirotetramat-enol is classified as highly mobile according to the classification scheme of McCall et al. (1981) 1 

### and mobile according to FAO 2 

### classification.

1 McCall, J.P., D.A. Laskowsky, R.L. Swann, and H.J. D ishburger. 1981. Measurement of sorption coefficien ts of organic chemicals and their use in environmental fate analy sis. Pages 89-109 In Test protocols for environmental fate & movement of toxicants. Proceedings of a symposium. Association of Official Analytical Chemists. 94 th Annual Meeting, October 21-22, 1980. Washington, DC.
2 Food and Agriculture Organization of the United Na tions. FAO PESTICIDE DISPOSAL SERIES 8. Assessing Soil Contamination: A Reference Manual. Appendix 2. Paramet ers of pesticides that influence processes in the s oil. Editorial Group, FAO Information Division: Rome, 2000.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reviewer comments:

• The notifier also stated K OC 

### values for the ‘immobile’ spirotetramat-enol fract ion in a range of 828 – 1711 L kg -1 

### . Since this fraction is considered to be strongly bound to the soil matrix via chemisorption, the ‘immobile’ fraction is considere d immobile per se . Partial transfer into deeper soil layers (‘Molino’ soil) may be attributed to (revers ible) binding of spirotetramat-enol onto more mobil e soil compounds (e.g. dissolved organic carbon, etc. ). Calculation of K OC 

### values for this tightly bound
fraction is not considered admissible at all. These data are not considered relevant for further risk assessment. (It may be annotated, that the calculat ion used by the notifier does not result in reliabl e K OC 

### values in the case that no transfer of the test it em into deeper soil layers is observed. The
calculation of the K OC 

### is solely based on the soil depth of maximum occur rence. Since this soil depth can only be very roughly estimated within the 0 – 6 cm column segment, calculated K OC 

### values are highly indicative and vary considerable assuming a depth of maximum occurrence of e.g. 1 or 3 cm).

• The reviewer calculated additional K OC 

### values for spirotetramat-ketohydroxy and spirotetr amat-MAamide on the basis of the column leachate data, in order to verify the reliability of K OC 

### results obtained for spirotetramat-enol. As can be seen in the following table, data from batch equilibrium experiments (Hein, 2005a; Simmonds and Early, 2005) and from the column study are highly consistent in case of spirotetramat-ketohydroxy and spirotetramat-MA-amide. In this respect, K OC 

### values obtained for spirotetramat-enol from the col umn study are considered reliable.

Table B.8.2.1.4-6: Calculated K OC 

### values for spirotetramat-ketohydroxy and spirotetr amat-MA-amid on the basis of column leachate data and results ob tained from batch equilibrium experiments – additional reviewer assessment.

V p [mL] a V v [mL] W [g] K d [L kg -1 ] K OC /K FOC [L kg -1 ] Compound Soil Column study Column study Batch

study b

‘Laacherhof AXXa’ 940 334 669 0.91 45 41.0
‘Laacherhof AIIIa’ 650 308 710 0.48 40 46.9
‘Hoefchen’ 990 368 583 1.07 44 41.2Spirotetramatketohydroxy

‘Molino’ 760 266 836 0.59 74 99.1

‘Laacherhof AXXa’ 440 334 669 0.16 7.9 4.4
‘Laacherhof AIIIa’ 560 308 710 0.35 30 6.5
‘Hoefchen’ 620 368 583 0.43 18 5.0SpirotetramatMA-amide

‘Molino’ 540 266 836 0.33 41 25.5

V p = Volume of water required to leach 50 % of the te st compound through the soil column (including void volume) [mL] (estimated by the reviewer) V v = Void volume of the column [mL] (mean values) W = Soil dry weight in the column [g] (mean values)

b Hein (2005a), Simmonds and Early (2005)



### Reference: Analysis of the sorption behaviour of spirotetramat -enol using a column leaching experiment Author(s), year: Hammel, K., 2007 Study/Report number: -, MEF-07/220 Guideline(s): Calculations – not applicable GLP: Calculations – not applicable Deviations: Calculations – not applicable Validity: Study considered as additional information, only – study not essential

Material and methods: The transport model HYDRUS was used to simulate the behaviour of spirotetramat-enol in a column study (Babczinski, 2006c) conducted on four soils. Three contrasting approaches were investigated:

• Equilibrium sorption model using the K OC 

### of the strongly sorbed spirotetramat-enol

### – 145 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

• Equilibrium sorption model using the K OC 

### of the mobile spirotetramat-enol

• Non-equilibrium sorption model assuming kinetic ad sorption for spirotetramat-enol (as used for the environmental risk assessment), input parameter wer e optimized by inverse modelling

For the two equilibrium sorption approaches (high and low K OC 

### ), degradation of spirotetramat-enol was considered to follow DFOP kinetics with a rapid (mo bile) and a slow degrading (strongly bound) fractio n. Adequate rates and sizes for the slow and the fast degrading fraction were derived from Röpke (2006a, Table B.8.4.4-7).

Table B.8.2.1.4-7: DFOP parameter derived from Röpk e (2006a) for the degradation of spirotetramatenol in soil used for the equilibrium sorption appr oaches.

Parameter Explanation / Calculation ‘Laacherhof AXXa’ ‘Laacherhof

AIIIa’ ‘Hoefchen’ ‘Molino’

g [0 ... 1] Size of the fast degrading (mobile)

fraction 0.851 0.833 0.888 0.863

1 – g [0 ... 1] Size of the slow degrading (strongly

bound) fraction 0.149 0.167 0.112 0.137

k 1 [days -1 ] Degradation rate of the fast (mobile)

degrading fraction 40.5 5.2 41.8 15.6

k 2 [days -1 ] Degradation rate of the slow

degrading (strongly bound) fraction 0.0098 0.0304 0.0108 0.0109



### For the non-equilibrium sorption approach , input data required in HYDRUS (i.e. Freundlich ad sorption distribution coefficient in the total system, K f 

### [days -1 

### ]; fraction of equilibrium sorption sites, f [0 ... 1 ]; degradation rate constant in the water, k w 

### [days -1 

### ]) were optimized for each soil column by inverse m odelling (using the iterative optimization algorithm of the computer program PEST). Optimization of input data was run until adequate fit between measured and modelle d amounts in the soil column and in the leachate fractions (weighted inversely proportional to the m ean of the observation group) was achieved. The kin etic sorption constant (k d 

### [days -1 

### ]) was directly taken from the SFO-RB modelling app roach (Röpke, 2006a).

Owing to the close conceptual analogy between HYDRU S and PEARL, the HYDRUS input parameters can be directly converted into corresponding PEARL para meters (for calculations refer to Table B.8.4.4-10)

.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.2.1.4-8: Particle density, dry bulk densi ty (Babczinski, 2006c) and calculated porosity and gravimetric water concentration of the soil column at saturation.

Parameter Explanation / Calculation ‘Laacherhof AXXa’ ‘Laacherhof

AIIIa’ ‘Hoefchen’ ‘Molino’

ρ r [g cm -3 ] Particle density 2.59 2.62 2.45 2.63

ρ s [g cm -3 ] Dry bulk density 1.13 1.23 0.61 0.46

Φ [-] Porosity, Φ = 1 – ρ s / ρ r 0.56 0.53 0.61 0.43

θ g [-] Gravimetric water content at saturation,

θ g = ( Φ × ρ w ) / ρ s,
ρ w = density of the water (i.e. 1 g cm -3 ) 0.50 0.43 0.63 0.32



### Table B.8.2.1.4-9: Optimized input parameter (after inverse modelling) of spirotetramat-enol in column studies modelled with HYDRUS (coupled to PES T).

Parameter Explanation ‘Laacherhof AXXa’ ‘Laacherhof

AIIIa’ ‘Hoefchen’ ‘Molino’ Remarks

k d [days -1 ] Kinetic sorption rate

constant 0.012 0.037 0.012 0.013 Values taken from Röpke (2006a)

K f [L kg -1 ] Freundlich adsorption

distribution coefficient (total system) 18.6 6.39 24.0 22.9 Optimized

f [0 ... 1] Fraction of equilibrium

sorption sites 0.036 0.14 0.039 0.050 a Optimized

k w [days -1 ] Degradation rate

constant in the water phase 2.40 2.63 2.14 3.60 Optimized

a Fixed



### Table B.8.2.1.4-10: Calculated PEARL parameter acco rding to optimized HYDRUS input parameter.

Parameter Explanation / Calculation ‘Laacherhof AXXa’ ‘Laacherhof

AIIIa’ ‘Hoefchen’ ‘Molino’

f ne [-] Ratio between non-equilibrium and

equilibrium sorption sites, f ne = (1 – f) / f 26.9 6.28 24.7 19.0

K f eq [L kg -1 ] Freundlich adsorption distribution

coefficient in the equilibrium domain, K f eq = K f / (1 + f ne ) 0.67 0.88 0.93 1.14

k t [days -1 ] Degradation rate constant in the

equilibrium phase, k t = k w / (1+ K f eq × ρ w

/ θ g ) 1.03 0.87 0.86 0.78

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Findings:

Figure B.8.2.1.4-1: Modelled and measured depth dis tribution (top within a soil) and breakthrough curve (bottom within as soil) of spirotetramat-enol for the column leaching in soils ‘Laacherhof AXXa’ (left, top), ‘Laacherhof AIIIa’ (right, top), ‘Hoefchen’ (left, bottom) and ‘Molino’ (right, bot tom).

### – 148 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion: The kinetic sorption model best reflects the measur ed values both in the eluate and distribution in th e soil column. Whereas the low equilibrium K OC 

### vastly overpredicts leachate concentrations, the h igh K OC 

### model underpredicts transport. The kinetic sorption model therefore offers the best description of the leach ing behaviour of spirotetramat-enol and is thus the bes t choice to extrapolate to natural field conditions

.

Reviewer comments:

• Study not considered essential. Study results included for additional information o nly.

• A comparison of the kinetic sorption data derived from the spirotetramat-enol degradation study applying SFO-RB kinetics (Babczinski, 2006b, Röpke, 2006a) and from inverse modelling of the column leaching study shows, that there are some di fferences in respect to the f ne 

### value (arithmetic mean 377 vs. 19.2) and the effective degradation ra te of spirotetramat-enol (geometric mean 16.6 vs. 2.64 days -1 

### ). However, the K OC 

### values of spirotetramat-enol, calculated from the column study (Babczinski, 2006c) and obtained by inverse modelli ng (this study) are almost consistent (Table B.8.2.4.1.-11). In general, one has to be careful c omparing the column study and the soil degradation study. In contrast to the soil degradat ion study, the column leaching study was conducted under conditions of high water saturation (slow oxygen supply, degradation of spirotetramat-enol is strongly driven by oxidation) and the experiment was conducted for only 5 days. In this respect, the reviewer agrees that the kinetic adsorption data obtained in the column study by inverse modelling are not adequate as mode lling input parameter for environmental risk

assessment.– 149 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.2.1.4-11: Comparison of kinetic sorptionparameters required in PEARL, derived from degradation rate study (spirotetramat-enol as parent,Babczinski, 2006b; Röpke, 2006a) and from the column study (this study).Parameter Explanation Study value deriving from‘LaacherhofAXXa’‘LaacherhofAIIIa’‘Hoefchen’ ‘Molino’ MeanColumn study (inverse modelling, HYDRUS) 33.5 73.338.8 142.572(geom.)KOC[L kg-1] KOCof the mobile fractionColumn study (calculated, Babczinski, 2006c) 27 6529 9955(geom.)Column study (inverse modelling, HYDRUS) 26.9 6.2824.7 19.019.2(arith.)fne[-]Ratio between non-equilibriumand equilibrium sorption sitesSoil degradation rate study (SFO-RBapproach, Röpke, 2006a)693 28.9 594 193377(arith.)Column study (inverse modelling, HYDRUS) 2.41 2.652.13 3.562.64(geom.)keff[days-1]Effective degradation rate of themobile fraction,keff= kt× (1 + Ke eq×ρw/θg)Soil degradation rate study (SFO-RBapproach, Röpke, 2006a)34.5 4.35 37.1 13.516.6(geom.)

### – 150 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.2.1.5 Aged residue column leaching studies (OEC D Annex IIA, 7.4.5)

Aged column leaching studies were not performed. Th is requirement is covered by the adsorption/desorpt ion studies with the parent compound and metabolites as presented in B.8.2.1.

B.8.2.1.6 Leaching (TLC) (OECD Annex IIA, 7.4.6)

Such data describing the mobility in soil were supp lied by higher tier studies shown in all other sect ions of point B.8.2.

B.8.2.1.7 Lysimeter studies (OECD Annex IIA, 7.4.7, Annex IIIA, 9.3.2)

The mobility of the spirotetramat residues in soil has been adequately assessed on the basis of the adsorption/desorption studies with the parent compo und and metabolites (as presented in B.8.2.1).

B.8.2.1.8 Field leaching studies (OECD Annex IIA, 7 .4.8, Annex IIIA, 9.3.3)

Based on the results of laboratory and modelling st udies mentioned in the chapter before it is conclud ed that the mobility of spirotetramat residues in soil is s ufficiently understood after its intended use, and no concern with regard to groundwater contamination is indicat ed. This was supported by the terrestrial field dis sipation studies performed in the USA, since there was no mo vement measured below a 15-cm depth of soil. Thus, field leaching studies were considered not necessar y.

B.8.2.1.9 Volatility laboratory and field study (OE CD Annex IIA, 7.4.9, IIIA, 9.3.4, 9.3.5)

The vapour pressure and Henry’s law constant of spi rotetramat were determined to be 5.6 × 10 -9 

### Pa (20 °C)
and 7.0 × 10 -8 

### Pa m 3 

### mol -1 

### (20 °C, pH 7), respectively. Based on these result s it was concluded that significant volatilization of spirotetramat in the environment is not to be expected. Therefore, no fu rther laboratory experiments were considered necessary.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.2.1.10 Summary on the mobility in soil

Note: In contrast to the EU risk assessment, the Canadia n and US risk assessment is based on linear (nonFreundlich) adsorption isotherms, and deviating inp ut data are used for calculation and modelling of predicted/expected environmental concentrations (e. g. the 20 th 

### percentile, or the lowest, median value, etc.). Respective values are stated in the summary tables together with the endpoints considered relevant for the
EU risk assessment.

Reliable adsorption constants according to Freundli ch isotherms (equilibrium batch experiments) could be
achieved for spirotetramat

, spirotetramat-ketohydroxy and spirotetramat-MA-amide using 5 soils from the EU, USA and Canada with a representative set of soil properties:

• Soil pH (CaCl 2 

### ): 4.7 – 6.8

• Organic carbon: 0.7 – 2.4 %

• Clay: 7 – 28 %

No adsorption/desorption constants are available fo r spirotetramat and metabolites in more alkaline so ils (pH ≥ 7.0 in CaCl 2 

### ). However, no dependency of the adsorption behavio ur onto the soil pH could be stated in the pH range tested. In this respect, it is unlikel y that adsorption/desorption under more alkaline co nditions will significantly differ from values obtained in t he tested range. This assumption is also confirmed by modelling results from the US field trials, showing a topsoil pH in the range of 6.3 – 8.1 (in water). These field trials were fully re-modelled with the (uncalibrate d) parameter set used for spirotetramat and metabol ites,
based on K OC 

### values obtained from the tested pH range of 4.7 – 6.8 (for details refer to Röpke and Ramanarayanan, 2007). Modelled and measured data we re highly consistent also in more alkaline soils.

Linear adsorption constants (K d 

### ) and Freundlich adsorption constants (K F 

### ) of spirotetramat have been determined in batch equilibrium experiments with fi ve different soils using the [3- 14 

### C] labelled test substance. Since significant degradation of the test item was observed in a pre-test, the main test was performed with sterilized soil (using mercuric(II)chloride) and th e equilibrium time was restricted to 3 hrs. Based o n organic carbon content, K FOC 

### values for the different soils were in a range of 159 – 435 L kg -1 

### (arithmetic mean 1/n = 0.94). Linear adsorption constants (K OC 

### ) were close to the Freundlich adsorption constants with a range of 184 – 437 L kg -1 

### . Based on these values, spirotetramat is classifie d as medium mobile according to the classification scheme of McCall et al. (1981) 1 

### and moderately mobile according to FAO 2 

### classification.

Adsorption behaviour of spirotetramat-ketohydroxy was investigated in the same soils tested with the parent, which were once again sterilized using merc uric(II)chloride to inhibit extensive degradation o f the test compound. Equilibration time was 24 hrs. Obtained K FOC 

### values were in a range of 41.0 – 99.1 L kg -1



### . Respective arithmetic mean 1/n value was 0.92. Line ar adsorption constants (K OC 

### ) were close to the Freundlich adsorption constants with a range of 42. 0 – 99.7 L kg -1 

### . Based on these values, spirotetramatketohydroxy is classified as highly mobile accordin g to the classification scheme of McCall et al. (19 81) and mobile according to FAO classification.

No significant breakdown of spirotetramat-MA-amide was observed in soil equilibrium experiments,
obtained K FOC 

### values were in a range of 4.4 – 25.5 L kg -1 

### . Respective arithmetic mean 1/n value was 0.95.

1 McCall, J.P., D.A. Laskowsky, R.L. Swann, and H.J. D ishburger. 1981. Measurement of sorption coefficien ts of organic chemicals and their use in environmental fate analy sis. Pages 89-109 In Test protocols for environmental fate & movement of toxicants. Proceedings of a symposium. Association of Official Analytical Chemists. 94 th Annual Meeting, October 21-22, 1980. Washington, DC.
2 Food and Agriculture Organization of the United Na tions. FAO PESTICIDE DISPOSAL SERIES 8. Assessing Soil Contamination: A Reference Manual. Appendix 2. Paramet ers of pesticides that influence processes in the s oil. Editorial Group, FAO Information Division: Rome, 2000.

### – 152 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Linear adsorption constants (K OC 

### ) were close to the Freundlich adsorption constants with a range of 4.2 – 25.6 L kg -1 

### . Based on these values, spirotetramat-MA-amide is classified as very highly mobile according to the classification scheme of McCall et al. (1981) a nd highly mobile according to FAO classification.

Owing to the extreme instability of spirotetramat-enol once in contact with soil, no reliable adsorption value
could be determined for this metabolite in a standa rd batch equilibrium experiment according to guidel ine OECD 107. The rapid disappearance of spirotetramat- enol during these tests is mainly attributed to fas t transformation to spirotetramat-ketohydroxy (and ot her degradation products) and to strong binding to the soil matrix (chemisorption), which is commonly obse rved in aerobic soils for this metabolite. In an ad ditional study to investigate the time-depending sorption, a dsorption and binding of spirotetramat-enol to soil was extremely fast and rapidly increased with aging tim e in soil. The fraction of spirotetramat-enol not t ightly bound to the soil, i.e. the fraction that can be re leased by aqueous solutions from soil (mobile fract ion), is degraded within a few hours (mainly to spirotetrama t-ketohydroxy). From this study, it can be conclude d that spirotetramat-enol is more or less absent from the soil pore water (either degraded or tightly bound t o soil) within a very short period of time. To obtain more reliable adsorption parameter for spirotetramat-eno l, a column study using spirotetramat-enol as parent was conducted on four soils (the same soils which were used for the spirotetramat-enol degradation study). From this study, it is evident that only a small f raction of spirotetramat-enol had a potential to leach into de eper soil layers or leave the soil column ( ≤ 2.8 % of AR in leachates), whereas by far the major fraction stron gly bound to the top soil layer. Only a minor part of the applied spirotetramat-enol (i.e. the mobile fractio n) was recovered in soil extracts and even a smalle r fraction in the leachate, indicating immediate and strong bi nding to the top soil, transformation into spirotet ramat-enol derived degradates, and significant formation of NE R during the progress of the study. For the mobile spirotetramat-enol fraction K OC 

### values between 27 – 99 L kg -1 

### were calculated. In this respect, spirotetramatenol is classified as highly mobile according to th e classification scheme of McCall et al. (1981) and mobile according to FAO classification. However, it should be kept in mind that this K OC 

### value is only valid for the mobile and fast degrading fraction of spirotetramat -enol (representing the first phase of the distinct bi-phasic degradation); the slowly degrading and strongly bou nd fraction of spirotetramat-enol (representing the second phase of the biphasic degradation) is consid ered more or less immobile.

In order to prove, whether the kinetic adsorption a pproach, used for spirotetramat-enol in the environ mental risk assessment, adequately describes the behaviour of the compound observed in the column leaching study, the column experiments were fully re-modelle d using the program HYDRUS. Several input parameters, among them the K OC 

### of spirotetramat-enol, were optimized by inverse m odelling, using the program PEST, in order to achieve adequate fits bet ween measured and modelled residues in the column and in the leachates. Based on these optimization p rocedures, the K OC 

### of spirotetramat-enol was calculated to be within a range of 34 – 143 L kg -1 

### , which is close to the K OC 

### values derived from more straightforward calculations based on the amounts of spirotetramat- enol in the leachate fractions, only.

According to the HPLC method, spirotetramat-enol dimer 1 and spirotetramat-enol dimer 2 show K OC 

### values of approx. 1624 L kg -1 

### and 3208 L kg -1 

### . Spirotetramat-enol dimer 1 is classified as lowly mobile according to the classification scheme of McCall et al. (1981) and slightly mobile according to FAO classification, spirotetramat-enol dimer 2 is class ified as slightly mobile according to the classific ation scheme of McCall et al. (1981) and according to FAO classification. However, neither spirotetramat-eno l dimer 1 nor spirotetramat-enol dimer 2 is considere d relevant for further risk assessment.– 153 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.2.1.10-1: Summary on adsorption constantsof spirotetramat and metabolites.Non-linear isotherm (Freundlich) Linear isothermCompound SoilpH(CaCl2)Corg[%]Clay[%]KF(ads)[L kg-1]KFOC(ads)[L kg-1]1/n R2Kd(ads)[L kg-1]KOC(ads)[L kg-1]R2pH depen-dencyMethod‘Laacherhof AXXa’ 6.1 2.38 7.0 4.79 201 1.001 0.9804.38 184 0.988‘Laacherhof AIII’ 6.8 0.87 16.7 3.78 435 0.892 0.993 3.80 437 0.972‘Hoefchen’ 5.9 2.33 15.0 4.10 176 0.945 0.994 4.69201 0.997‘Molino’ 5.4 0.93 5.4 4.05 435 0.823 0.976 3.58 3850.809‘Saskatoon’ 4.7 2.33 24.4 3.70 159 1.042 0.903 5.52237 0.914Arithmetic mean (EU)---nc2810.941----Spirotetramat20th%ile (PMRA)-------3.76198-No Batch equilibrium‘Laacherhof AXXa’ 6.1 2.0 10 - - 1.00a- 0.54 27 -‘Laacherhof AIII’ 6.5 1.2 20 - - 1.00a- 0.78 65 -‘Hoefchen’ 6.4 2.4 18 - - 1.00a- 0.70 29 -‘Molino’ 5.4 0.8 10 - - 1.00a- 0.79 99 -Arithmetic mean (EU)---nc-1.00--55-Spirotetramatenol20th%ile (PMRA)-------0.6428-No Column study‘Laacherhof AXXa’ 6.0 1.30 8.9 0.533 41.0 0.920 1.000 0.560 43.1 0.998‘Laacherhof AIII’ 6.4 1.10 13.2 0.516 46.9 0.929 1.000 0.529 48.1 0.997‘Hoefchen’ 6.1 2.62 14.7 1.08 41.2 0.927 1.000 1.1042.0 0.996‘Molino’ 6.2 0.87 8 0.862 99.1 0.918 1.000 0.867 99.7 0.994‘Saskatoon’ 5.5 2.44 28 2.21 90.4 0.915 1.000 2.2893.4 0.994Arithmetic mean (EU)---nc63.70.922----Spirotetramatketohydroxy20th%ile (PMRA)-------0.5542.9-No Batch equilibrium‘Laacherhof AXXa’ 6.3 1.70 8.9 0.075 4.4 1.07 1.0000.071 4.2 0.997‘Laacherhof AIII’ 6.7 0.91 14.9 0.059 6.5 0.96 0.999 0.064 7.0 0.998‘Hoefchen’ 6.5 2.07 14.7 0.104 5.0 0.80 0.998 0.1165.6 0.976‘Molino’ 5.7 0.7 7 0.179 25.5 0.98 0.999 0.179 25.60.997‘Saskatoon’ 5.4 2.3 25 0.117 5.1 0.93 0.999 0.126 5.5 0.994Arithmetic mean (EU)---nc9.30.95----Spirotetramatketohydroxy20th%ile (PMRA)-------0.0705.2-No Batch equilibriumSpirotetramatenol dimer 1b1624nd HPLC methodSpirotetramatenol dimer 2b3208nd HPLC methodnd denotes not determinableaUnknown, set to 1.00 to adequately account for theSFO-RB approach (more conservative than the FOCUSdefault of 0.9)bSpirotetramat-enol dimer 1 and 2 not considered relevant for further risk assessment (artefacts owingto hot spot application in the lab)

### – 154 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.3 Predicted environmental concentration in soil (PEC S 

### ) (OECD Annex IIIA, 9.4, 9.5)

The risk assessment for the EU can be found in the APPENDIX.
The risk assessment for the PMRA and the US-EPA is not included in this monograph.

The Proposed Registration Decision – Spirotetramat (PRD2008-07), containing the detailed Canadian review and risk assessment, has been published by t he PMRA and can be found at the PMRA’s web site www.pmra-arla.gc.ca

.

The registration decision made by the U.S. EPA's Of fice of Pesticide Program is supported by the risk
assessments found at our Federal Docket website loc ated at: http://www.regulations.gov/fdmspublic/component/mai n?main=AdvancedDocket
The Docket ID is "EPA-HQ-OPP-2007-0475".

### – 155 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.4 Fate and behaviour in the water

B.8.4.1 Hydrolytic degradation (OECD Annex IIA, 7.5 )

Studies on the hydrolytic degradation were conducte d with

• Spirotetramat (pH 4, 7 and 9), [3- 14 

### C] and [5- 14 

### C] label

• Spirotetramat-enol (pH 4, 7 and 9), [3- 14 

### C] and [5- 14 

### C] label

Reference: [Azaspirodecenyl-3- 14 

### C]- and [Azaspirodecenyl-5- 14 

### C]BYI08330: Hydrolytic degradation Author(s), year: Heinemann, O., 2004a Study/report number: M-09312401-2, MEF-04/176 Guideline(s): 94/37/EC, 95/36/EC, OECD Guideline 11 1 (2002), US-EPA N 161-1 (1982), Canada PMRA DACO 8.2.3.2 (1987), Japan MAFF New Tes t Guideline 12 Nousan 8147 (2001)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods:
Test substance: • [Azaspirodecenyl-3- 14 

### C]-spirotetramat, 3.71 MBq mg -1



### , > 99 % radiochemical purity (HPLC), batch BECH 0980 and BECH 1518

• [Azaspirodecenyl-5- 14 

### C]-spirotetramat, 4.03 MBq mg -1



### , > 98 % radiochemical purity (HPLC), batch BECH 0981 and BECH 1519 Labels regarded as replicates Reference substances: Spirotetramat (unlabelled), S pirotetramat-enol, Spirotetramat-ketohydroxy Test systems: • pH 4: 0.01 M sodium acetate/acetic acid buffer (ad justed with acetic acid)

• pH 7: 0.01 M tris(hydroxymethyl)aminomethane buffe r (adjusted with 0.1 M HCl and/or 0.1 M NaOH)

• pH 9: 0.01 M borate buffer (adjusted with 0.4 M Na OH and/or boric acid) All buffers sterilized (steam pressure sterilizatio n) and checked for sterility throughout the test duration. Oxygen content reduce d by nitrogen bubbling. Volatile traps: No volatile traps (no volatiles exp ected – confirmed by complete material balance) Test temperature: 50 °C (pre-test), 30 °C (pre-test , pH 9 only), 25 °C (main test) and 20 °C (optional test, pH 4 and 7 only) Test duration: From 7 to 48 days in the dark Sample concentration: 1.0 mg L -1 

### Co-solvent: Acetonitrile (< 0.5 %, v/v) Analysis: LSC, HPLC-UV/RAD, TLC, HPLC-MS/MS (ESI)
LOQ < 1 % of AR Kinetic evaluation: Simple first order (SFO) kineti cs, ModelManager 1.1, curve fit based on mean
values of both labels

Findings: Mean material balances of all ( n = 8) experiments were in a range of 97.2 – 104.4 % of AR. Owing to the complete mass balance 14 

### CO 2 

### formation is considered to be negligible. Spirotet ramat showed distinct hydrolysis at all temperatures and pH value tested, hydrolysis increased with increasing temperature a nd increasing alkalinity. No distinct differences betw een labels tested occurred. At 25 °C calculated hal f-life times were 32.5, 8.6 and 0.32 days (all SFO, R 2 

### > 0.99) at a pH of 4, 7 and 9. The only major meta bolite

### – 156 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

formed in all tests was the common hydrolysis produ ct spirotetramat-enol, which can be considered to b e stable under the experimental conditions. Maximum o f one unknown peak area was 8.8 % of AR (only using
5- 14 

### C labelled parent at 50 °C and at a pH of 4). Howev er, this peak area is considered as artefact, proba bly due to sample autoxidation prior to chromatography. Unidentified radioactivity did not exceed 2.7 % of AR.– 157 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.4.1-1: Hydrolysis of spirotetramat in sterile water buffered at pH 4, 7 and 9 [% of AR].pH479Temperature50 °C 25 °C 20 °C 50 °C 25 °C 20 °C 30 °C 25 °CCompound/LabelDAT [3-14C][5-14C]DAT [3-14C][5-14C]DAT [3-14C][5-14C]HATa[3-14C][5-14C]DAT [3-14C][5-14C]DAT [3-14C][5-14C]HAT [3-14C][5-14C]HAT [3-14C][5-14C]0100.0 100.00100.0 100.00100.0 99.20100.0 100.00100.0 99.50100.0 100.0097.0 97.3098.0 97.60.2599.9 98.0396.4 94.1591.5 91.7290.4 88.8189.8 90.9577.5 75.2175.9 79.4187.2 87.7371.3 69.6786.4 85.7888.3 89.1482.8 81.9377.4 79.5865.5 65.3261.4 65.5279.0 81.0462.6 60.31079.6 79.41382.3 83.4868.9 69.2755.7 56.01350.2 48.5350.6 53.0372.4 73.3554.7 52.31474.7 74.01678.6 78.91061.8 63.41335.8 35.31642.0 44.5441.0 41.7464.9 65.4651.0 48.61769.8 68.01976.0 75.52432.1 32.62019.2 19.81936.6 35.8533.4 31.5655.6 58.1744.0 43.42165.4 64.42369.8 71.73025.0 24.92415.0 14.02328.7 30.3628.3 26.3847.3 45.91030.2 29.82460.7 59.42667.2 68.74810.2 11.02910.3 8.92626.4 25.0721.3 22.310na 38.71223.2 22.43152.9 51.03063.5 64.83020.4 20.5249.9 12.5Spirotetramat30na 7.10nd nd0nd nd0nd nd0nd nd0nd nd0nd nd03.0 2.302.0 2.40.253.0 3.536.3 6.957.2 7.029.611.219.6 9.3524.025.0121.617.5111.711.8329.130.0714.014.0810.910.8418.618.7323.023.1836.335.8236.631.9220.620.3439.838.11019.218.91317.616.7833.532.4744.545.41351.853.1347.842.9327.827.9543.543.91426.125.41621.020.31040.538.51365.266.51660.457.2457.556.9435.034.6651.552.61730.430.31923.924.82472.170.12082.883.01965.564.0565.264.5643.544.2758.752.92136.036.52327.427.63079.577.82484.187.52372.371.4670.868.1852.355.81071.452.22438.338.82631.030.64893.493.4 2991.492.2 2675.473.8775.475.9 10na64.11275.469.3 3149.847.8 3035.033.93081.179.92490.089.6Spirotetramatenol30na92.6nd denotes not determinedna denotes not analysedaHAT denotes hours after treatment.

### – 158 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.4.1-2: Calculated hydrolytic DT 50 

### and DT 90 

### of spirotetramat in sterile water buffered at pH 4
, 7 and 9 using SFO kinetics (mean of both labels). pH Temp [°C] DT 50 [days] DT 90 [days] R 2 Kinetics

50 5.69 18.9 1.00 SFO
25 32.5 108 1.00 SFO 4

20 47.6 158 1.00 SFO

50 0.62 2.08 1.00 SFO
25 8.6 28.7 1.00 SFO 7

20 13.1 43.5 1.00 SFO

30 0.14 0.45 1.00 SFO 9 25 0.32 1.04 1.00 SFO



### Conclusion: At environmental relevant conditions spirotetramat is degrading with a half-life of 32.5, 8.6 and 0.32 days at pH 4, 7 and 9 and 25 °C. It is concluded that hydro lysis is relevant for the overall elimination of sp irotetramat in aquatic systems, especially under neutral and al kaline conditions. The only major metabolite derivi ng from hydrolysis of spirotetramat is spirotetramat-enol w hich is considered to be stable under the experimen tal
conditions.

Reviewer comments:
None

Reference: [Azaspirodecenyl-3- 14 

### C]- and [Azaspirodecenyl-5- 14 

### C]BYI08330-Enol: Hydrolytic degradation Author(s), year: Heinemann, O., 2004b Study/report number: M-242999-01-2, MEF-04/311 Guideline(s): 94/37/EC, 95/36/EC, OECD Guideline 11 1 (2002), US-EPA N 161-1 (1982), Canada PMRA DACO 8.2.3.2 (1987), Japan MAFF New Tes t Guideline 12 Nousan 8147 (2001)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods:
Test substance: • [Azaspirodecenyl-3- 14 

### C]-spirotetramat-enol, 4.54 MBq mg -1



### , > 99 % radiochemical purity (HPLC), batch BECH 1564

• [Azaspirodecenyl-5- 14 

### C]-spirotetramat-enol, 4.99 MBq mg -1



### , > 99 % radiochemical purity (HPLC), batch BECH 1565 Labels regarded as replicates Reference substances: Spirotetramat-enol (unlabelle d) Test systems: • pH 4: 0.01 M sodium acetate/acetic acid buffer (ad justed with acetic acid)

• pH 7: 0.01 M tris(hydroxymethyl)aminomethane buffe r (adjusted with 0.1 M HCl and/or 0.1 M NaOH)

• pH 9: 0.01 M borate buffer (adjusted with 0.4 M Na OH and/or boric acid) All buffers sterilized (steam pressure sterilizatio n) and checked for sterility throughout the test duration. Oxygen content reduce d by nitrogen bubbling. Volatile traps: No volatile traps (no volatiles exp ected – confirmed by complete material balance) Test temperature: 50 °C (pre-test) and 25 °C (main test) in the dark Test duration: 5 days (pre-test) and 31 days (main test) Sample concentration: 1.0 mg L -1

### – 159 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Co-solvent: Methanol/aqueous 0.01 M NaHCO 3 

### (1/1, v/v), < 0.3 % of total volume Analysis: LSC, HPLC-UV/RAD, TLC, HPLC-MS/MS (ESI)
LOD < 1 % of AR Kinetic evaluation: Simple first order (SFO) kineti cs, ModelManager 1.1, curve fit based on mean
values of both labels

Findings: Material balance was in the range of 97.5 – 102.6 % of AR in both experiments for both labels used. No timedependent degradation of spirotetramat-enol was obs erved, spirotetramat-enol (mean of both labels) accounted for 98.2 – 101.2 % of AR throughout the p re-test and 98.0 – 100.5 % of AR in the main test. No single radioactive species other than the parent te st item occurred at levels > 2.0 % of AR.

Conclusion: Spirotetramat-enol is hydrolytically stable under a biotic environmental conditions. Hydrolysis is not expected to contribute to the degradation of spirotetramat-e nol in the environment.

Reviewer comments:
None

### – 160 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.4.2 Photolytic degradation (OECD Annex IIA, 7.6 )

Studies on the photolytic degradation were conducte d with spirotetramat in

• sterile buffered water (pH 5.0), [3- 14 

### C] and [5- 14 

### C] label

• sterile natural water (pH 7.9), [3- 14 

### C] and [5- 14 

### C] label

Quantum yields and half-life time under environment al conditions were determined for:

• Spirotetramat

• Spirotetramat-enol

Reference: BYI08330: Phototransformation of BYI08330 in steril e water Author(s), year: Stupp, H.-P., 2005a Study/report number: M-266695-01-2, MEF-05/206 Guideline(s): 95/36/EC, 94/37/EC, SETAC (1995), US- EPA N 162-1 (1982), Canada PMRA DACO 8.2.3.3.2 (1987)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods:
Test substances: • [Azaspirodecenyl-3- 14 

### C]-spirotetramat, 3.67 MBq mg -1



### , > 98 % radiochemical purity (HPLC), batch BECH 0950

• [Azaspirodecenyl-5- 14 

### C]-spirotetramat, 4.03 MBq mg -1



### , ≥ 98 % radiochemical purity (HPLC), batch BECH 0952 Labels regarded as replicates Reference substances: Spirotetramat (unlabelled), S pirotetramat-enol Test system: Sterile pH 5.0 buffer (0.1 M acetate b uffer, autoclaved), adjusted with 0.1 M NaOH, sterility was checked throughout the experime nt Test temperature: 25 ± 1 °C Test duration: 7 days continuous irradiation (equiv alent to 51.7, 33.3 and 47.0 solar midsummer days in at Athens (Greece, EU, 38 °N), Phoenix (AZ, USA, 33 °N) and Edmonton (Alberta, CAN, 53 °N), respectively) or dark incuba tion Sample concentration: 1.0 mg L -1 

### Co-solvent: Acetonitrile (0.1 %, v/v) Test system: Xenon arc lamp (Xenotest), cut-off < 2 90 nm, 990 W m -2 

### (300 – 800 nm), 22 cm above test vessel, light intensity measured by an i rradiance sensor

### – 161 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Spectrum of experimental radiation is qualitatively similar to solar irradiation. Volatile traps: 1 x soda lime and 1 x polyurethane foam Analysis: LSC, HPLC-UV/RAD, TLC, HPLC-MS/MS (ESI), LC-NMR
LOD = 1 % of AR Kinetic evaluation: Simple first order (SFO) kineti cs, ModelManager 1.1, curve fit based on mean
values of both labels

Findings: Mass balance was in a range of 93.8 to 108.4 % of A R for all experiments. Under the impact of light sm all amounts (maximum 2.6 % of AR) of 14 

### CO 2 

### were released from the [3- 14 

### C] labelled parent indicating mineralization at this part of the molecule. Almost no 14 

### C was released from [5- 14 

### C] labelled spirotetramat. Formation of organic volatiles was negligible. Unde r irradiation spirotetramat was subjected to extens ive photolytic rearrangement procedures, resulting in 4 major metabolites: spirotetramat-photo-cyclopentyl (maximum 42.9 % of AR), spirotetramat-photo-hydroxy methyl (22.9 % of AR), spirotetramat-photo-formyl (11.5 % of AR) and spirotetramat-photomethyl carbon ate (19.3 % of AR). These major transformation products did not decline significantly. Unknown com pounds did not exceed 5.5 % of AR individually. Wit hout irradiation only the common hydrolysis product spir otetramat-enol was formed.– 162 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.4.2-1: Photo-transformation of spirotetramat in sterile water buffered at pH 5 [% of AR].LabelCondi-tionsDAT CO2OrganicvolatilesSpirotetramatSpirotetramat-enolSpirotetramat-photo-cyclopentylSpirotetramat-photo-hydroxymethylSpirotetramat-photo-formylSpirotetramat-photo-methylcarbonateIndividualmaximumof others(n = 5)Total0na na 100.2 0.0 0.0 0.0 0.0 0.0 0.0 100.210.1 0.1 72.9 0.0 3.116.9 0.0 7.2 0.0 100.220.2 0.0 64.0 0.0 5.818.4 2.8 9.5 0.0 100.930.8 0.1 36.4 0.019.216.210.714.0 5.1 102.541.3 0.0 34.6 0.024.922.24.916.1 0.0 104.062.6 0.0 16.8 0.037.615.8 3.519.34.8 105.7Irradiated72.5 0.8 16.7 0.035.415.5 6.317.05.2108.40na na 100.9 0.0 0.0 0.0 0.0 0.0 0.0 100.91na na 102.4 2.3 0.0 0.0 0.0 0.0 0.0 104.72na na 93.5 4.2 0.0 0.0 0.0 0.0 0.0 97.73na na 90.5 6.5 0.0 0.0 0.0 0.0 0.0 97.04na na 88.5 9.0 0.0 0.0 0.0 0.0 0.0 97.66na na 84.511.7 0.0 0.0 0.0 0.0 0.0 96.1[3-14C]Dark7na na 84.613.70.0 0.0 0.0 0.0 0.0 98.30na na 98.2 0.0 0.0 0.0 0.0 0.0 0.0 98.210.0 0.3 78.3 0.0 2.313.7 0.0 5.5 0.0 100.020.0 0.0 64.5 0.0 5.418.1 2.3 8.8 1.4 100.530.0 0.0 39.3 0.015.115.411.511.75.5103.540.1 0.0 41.1 0.016.621.9 3.914.7 5.3 103.560.1 0.0 32.7 0.025.322.93.617.2 4.4 106.3Irradiated70.2 0.1 12.0 0.042.913.8 4.318.45.2 106.00na na 101.4 0.0 0.0 0.0 0.0 0.0 0.0 101.41na na 97.5 2.6 0.0 0.0 0.0 0.0 0.0 100.12na na 97.3 4.2 0.0 0.0 0.0 0.0 0.0 101.63na na 86.8 6.9 0.0 0.0 0.0 0.0 0.0 93.84na na 89.5 9.8 0.0 0.0 0.0 0.0 0.0 99.46na na 89.112.5 0.0 0.0 0.0 0.0 0.0 101.6[5-14C]Dark7na na 85.113.80.0 0.0 0.0 0.0 0.0 98.9

### – 163 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.4.2-2: Calculated photolytic DT 50 

### and DT 90 

### [days] of spirotetramat in 0.01 M acetate buffer a t pH 5.0 using SFO kinetics (mean value of both label s).

Experimental Natural conditions

Conditions DT 50

[days] DT 90

[days] R 2 DT 50 [days],

Athens, Greece, EU DT 50 [days],

Phoenix, AZ, USA DT 50 [days],

Edmonton, Alberta, Canada Kinetics

Irradiated 2.7 9.0 0.98 12.0 a 9.3 a 11.4 a SFO
Dark 26.2 86.9 0.89 26.2 26.2 26.2 SFO
Net b 3.0 10.0 - 22.2 14.4 20.2 SFO

a Re-calculated by the reviewer (see comments)
b Calculated by the reviewer



### Conclusion: Under irradiated experimental conditions, spirotetr amat degraded with a half-life of 2.7 days at a pH of 5 and 25 °C (SFO kinetics, R 2 

### = 0.98) owing to photolysis and hydrolysis. This e xperimental half-life corresponds to 12.0, 9.3 and 11.4 days under environmental midday summer light in Athens (Greece, EU), Phoenix (AZ, USA) and Edmonton (Alberta, Canada). In the dark co ntrol DT 50 

### was 26.2 days (owing to hydrolysis) indicating that photodegradation of spirotetramat c an be considered a significant route for the elimin ation of the compound from the environment. Under the steril e experimental conditions and the impact of light, spirotetramat is transformed to the major metabolit es spirotetramat-photo-cyclopentyl, spirotetramat-p hotohydroxymethyl, spirotetramat-photo-formyl and spiro tetramat-photomethyl carbonate (all these metabolit es considered as photo-rearrangement products). Maximu m amounts formed were 42.9, 22.9, 11.5 and 19.3 % of AR, respectively. Under dark conditions only the common hydrolysis product spirotetramat-enol was
observed.

Reviewer comments:

• Labels were handled as replicates. Additional calc ulations by the reviewer confirmed that the degradation of both labels is almost identical (DT 50 

### [3- 14 

### C] = 2.5 days, [5- 14 

### C] = 2.9 days), therefore the usage of a mean value is considered acceptable.

• Based on the net photosynthesis half-life of 3.0 d ays (calculated by the reviewer), experimental netphotolysis half-life of spirotetramat at pH 5.0 cor responds to 22.2, 14.4 and 20.2 days under environmental midday summer light in Athens (Greece , EU), Phoenix (AZ, USA) and Edmonton (Alberta, Canada). For calculating the effective degradation half-life under environmental condition s (owing to photolysis and hydrolysis), corrected net -photolysis and non-corrected hydrolysis rates (expressed as k

rates) were summed up. Calculations conducted by t he notifier (including hydrolysis rate, which was also corrected) are not considered valid.

Reference: [Azaspirodecenyl-3- 14 

### C]BYI08330 and [azaspirodecenyl-3- 14 

### C]BYI08330:
Phototransformation in natural water Author(s), year: Stupp, H.-P., 2005b Study/report number: M-266753-01-2, MEF-05/262 Guideline(s): 95/36/EC, 94/37/EC, SETAC (1995), US- EPA N 162-1 (1982), Canada PMRA DACO 8.2.3.3.2 (1987)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods:
Test substances: • [Azaspirodecenyl-3- 14 

### C]-spirotetramat, 3.67 MBq mg -1



### , > 98 % radiochemical purity (HPLC), batch BECH 0950

• [Azaspirodecenyl-5- 14 

### C]-spirotetramat, 4.03 MBq mg -1



### ,

### – 164 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

> 98 % radiochemical purity (HPLC), batch BECH 0952 Labels regarded as replicates Reference substances: Spirotetramat (unlabelled), S pirotetramat-enol Test system: Sterilized (autoclaved) natural water sampled from the Rhine River: pH: 7.93 Suspended solids: 25.8 mg L -1 

### Total evaporation residue: 273.3 mg L -1 

### Oxygen saturation: 98.4 % at 16.5 °C Conductivity: 523 μS cm -1 

### TOC: 2 mg L -1 

### DOC: < 2 mg L -1



### Hardness: 10.6 °dH Total nitrate: 8.8 mg L -1


### Total phosphorous: 0.1 mg L -1


### UV-Absorption: < 0.05 (290 – 800 nm) Test temperature: 25 ± 1 °C Test duration: 10 days continuous irradiation (equi valent to 52.2, 33.7 and 47.5 solar midsummer days in at Athens (Greece, EU, 38 °N), Phoenix (AZ, USA, 33 °N) and Edmonton (Alberta, CAN, 53 °N), respectively) or dark incuba tion

Spectrum of experimental radiation is qualitatively similar to solar irradiation. Sample concentration: 1.0 mg L -1 

### Co-solvent: Acetonitrile (0.1 %, v/v) Test system: Xenon arc lamp (Xenotest), cut-off < 2 90 nm, approx. 700 W m -2 

### (300 – 800 nm), 22 cm above test vessel, light intensity measured b y an irradiance sensor Volatile traps: 1 x soda lime and 1 x polyurethane foam Analysis: LSC, HPLC-UV/RAD, TLC, HPLC-MS/MS, LC-NMR
LOD = 0.5 % of AR Kinetic evaluation: Simple first order (SFO) kineti cs, ModelManager 1.1, curve fit based on mean
values of both labels

Findings: Mass balance was in a range of 94.8 to 103.2 % of A R for all experiments. Under the impact of light sm all amounts (maximum 1.1 % of AR) of 14 

### CO 2 

### were released from the [3- 14 

### C] labelled parent. Almost no 14 

### C was

### – 165 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

released from [5- 14 

### C] labelled spirotetramat. Formation of organic vol atiles was negligible. Under irradiation in the sterile natural water spirotetramat was subj ected to extensive photolytic degradation into seve ral (n ≤ 33) metabolites, including spirotetramat-enol (maxi mum 81.9 % of AR, significantly decreasing thereaft er) and two further major metabolites, which were only observable using [5- 14 

### C] labelled spirotetramat: methoxycyclohexylaminocarboxylic acid (maximum 11.3 % of A R) and methoxycyclohexanone (17.5 % of AR), both showing maximum occurrence towards study end. One u nidentified peak area reached a maximum occurrence of 9.6 % of AR by 8 days decreasing ther eafter to 9.1 % of AR by 10 DAT. Further unknown compounds did not exceed 5.6 % of AR individually. Without irradiation only the common hydrolysis prod uct spirotetramat-enol was formed (complete hydrolysis of the parent).

Table B.8.4.2-3: Photo-transformation of spirotetra mat in sterile natural water at pH 7.9 [% of AR].

Label Conditions DAT CO 2 Organic volatiles Spirotetramat Spirotetramatenol PB1 a PB3 b Individual maximum of others ( n ≤ 33) Total

0 na na 80.4 16.3 0.0 96.7

1 0.0 0.0 2.6 78.3 2.0 103.2
2 0.0 0.0 0.0 68.0 2.7 103.2
3 0.1 1.5 0.0 56.5 4.0 103.2
6 0.3 0.7 0.0 32.7 7.7 102.9
8 0.7 1.0 0.0 21.5 9.6 102.7 Irradiated

10 1.1 0.7 0.0 14.1 9.1 102.5

0 na na 88.3 7.9 0.0 96.2

1 na na 61.7 37.7 0.0 99.3
2 na na 35.4 64.8 0.0 100.3
3 na na 21.9 79.1 0.0 101.0
6 na na 3.7 98.6 0.0 102.4
8 na na 1.5 100.8 0.0 102.3 3- 14 C

Dark

10 na na 0.0 102.7 Not detectable using this label

0.0 102.7

0 na na 77.5 19.3 0.0 0.0 0.0 96.8

1 0.0 0.0 1.5 81.9 0.6 3.0 2.0 103.2

2 0.0 0.0 0.0 72.2 1.5 6.0 2.9 103.2

3 0.0 0.1 0.0 58.3 2.7 8.1 4.1 102.4

6 0.1 0.1 0.0 32.1 6.1 15.0 5.2 101.9

8 0.2 0.1 0.0 17.0 9.9 17.5 5.2 102.5 Irradiated

10 0.3 0.1 0.0 16.8 11.3 16.0 5.6 102.5

0 na na 84.0 10.9 0.0 0.0 0.0 94.8

1 na na 57.5 40.9 0.0 0.0 0.0 98.4

2 na na 35.6 64.7 0.0 0.0 0.0 100.3

3 na na 20.9 78.9 0.0 0.0 0.0 99.8

6 na na 3.8 97.4 0.0 0.0 0.0 101.2

8 na na 1.5 100.8 0.0 0.0 0.0 103.0 5- 14 C

Dark

10 na na 0.0 102.0 0.0 0.0 0.0 102.0

a Methoxy-cyclohexylaminocarboxylic acid
b Methoxycyclohexanone

### – 166 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.4.2-4: Calculated photolytic DT 50 

### and DT 90 

### [days] of spirotetramat in sterile natural water a t pH 7.9 using SFO kinetics (mean value of both label s).

Experimental Natural conditions

Conditions DT 50

[days] DT 90

[days] R 2 DT 50 [days],

Athens, Greece, EU DT 50 [days],

Phoenix, AZ, USA DT 50 [days],

Edmonton, Alberta, Canada Kinetics

Irradiated 0.19 0.63 1.00 0.66 a 0.50 a 0.62 a SFO
Dark 1.54 5.11 1.00 1.54 1.54 1.54 SFO
Net b 0.22 0.71 - 1.15 0.74 1.05 SFO

a Re-calculated by the reviewer (see comments)
b Calculated by the reviewer



### Conclusion: Under irradiated experimental conditions, spirotetr amat extensively degraded with a half-life of 0.19 days in a sterilized natural water at pH 7.9 and 25 °C (SFO k inetics, R 2 

### = 1.00) owing to photolysis and hydrolysis. This experimental half-life corresponds to 0.66, 0. 50 and 0.62 days under environmental midday summer light in Athens (Greece, EU), Phoenix (AZ, USA) and Edmonton (Alberta, Canada). In the dark control, D T 50 

### was 1.54 days (owing to hydrolysis) indicating that photodegradation of spirotetramat can be considere d as a significant route for the elimination of the compou nd from the environment. Two major phototransformat ion products were formed besides the common hydrolysis product spirotetramat-enol: methoxycyclohexylaminocarboxylic acid (maximum 11.3 % of A R at study termination) and methoxycyclohexanone (17.5 % of AR). Under dark conditions only spirotet ramat-enol was formed owing to hydrolysis.

Reviewer comments:

• Based on the net photosynthesis half-life of 0.22 days (calculated by the reviewer), experimental net

photolysis half-life of spirotetramat at pH 7.9 cor responds to 1.1, 0.7 and 1.0 days under environmental midday summer light in Athens (Greece , EU), Phoenix (AZ, USA) and Edmonton (Alberta, Canada). For calculating the effective d egradation half-life under environmental conditions (owing to photolysis and hydrolysis), corrected net -photolysis and non-corrected hydrolysis rates (expressed as k

rates) were summed up. Calculations conducted by t he notifier (including hydrolysis, which was also corrected) are not considered valid.

• This photolysis study in sterile natural water was conducted to demonstrate that photorearrangement products observed under photolysis in sterile buffer solution at pH 5 (Stupp, 2005a) are not likely to occur under more natural conditio ns. However, based on the contrasting pH value of both photolysis studies (pH 5.0 in sterile buffer s olution vs. pH 7.9 in sterile natural water) no fin al conclusion on the occurrence of photo-rearrangement product in sterilized natural water in more acidic water can be drawn from the study results per se . However, based on the rapid degradation half-life (dark rate) of spirotetramat in water/sed iment systems (pH 6.7 and 7.2) of approx. 0.8 days (Menke, 2006b, Röpke, 2006b) a significant formatio n of photo-rearrangement products as observed in Stupp (2005a), in which spirotetramat showed a d ark hydrolysis rate of 26.2 days, is highly unlikely to occur.

### – 167 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: BYI08330: Determination of the quantum yield and as sessment of the environmental half-life of the direct photodegradat ion in water Author(s), year: Heinemann, O., 2004c Study/report number: M-092941-01-2, MEF-04/080 Guideline(s): Phototransformation of Chemicals in W ater, part A: Direct Phototransformation, UBA, Berlin, Germany (1992-12); Test Method: ECETOC (Polychromatic Light Source), US-EPA Guideline No. 161-2 SUPP
GLP: Yes
Deviations: None Validity: Study considered acceptable (see comments )

Material and methods: Light source: Mercury immersion lamp TQ 150 (cut-of f filter < 295 nm) Actinometer solution: Uranyl oxalate (0.01 mol L -1 

### uranyl ions, 0.05 mole L -1 

### oxalic acid) Exposure of actinometer solution: 10 min at 25 °C Test solution: Spirotetramat (unlabelled), 5.0 mg L -1 

### in Milli-Q-water (unbuffered) Exposure of test solution: 4 hrs Range of wave-lengths investigated: 295 – 490 nm Analysis of parent degradation: HPLC-UV UV/VIS-spectra of test substance: In test solution and in buffered solutions at pH 4, 7 and 9 (all 0.0 1 M, 10.0 mg test item L -1 

### ) Modelling of environmental half-life: a) GC-SOLAR b) Frank & Klöpffer

Findings: Based on actinometer (uranyl oxalate) degradation, a radiation intensity of approx. 6.80 × 10 16 

### photons (sec × 3 ml) -1 

### was calculated for the test unit. The UV/VIS absor ption spectrum of 10.0 mg L -1 

### spirotetramat in pure water shows a maximum below 210 nm and a sh oulder at approx. 218 nm, determination of absorption was measured above 490 nm. The UV/VIS sp ectra of spirotetramat in the buffered solutions showed comparable absorption properties. The degrad ation of spirotetramat under test conditions (unbuffered solution in water) accounted for approx . 56 % over an irradiation period of 4 hrs. Mean degradation half-life under the experimental condit ions was calculated to be 0.15 days (SFO kinetics). Based on the degradation and the adsorption data, this re sulted in a mean quantum yield of direct phototransformation in aqueous solutions of Φ = 0.00571.

The environmental half-life of spirotetramat, calcu lated according to GC-SOLAR using the quantum yield of

Φ = 0.00571 and the molar extinction coefficients fr om 297.5 to 490 nm were spread from 0.5 – 8.7 days depending on the latitude and the season.

Table B.8.4.2-5: GC-SOLAR modelling: Environmental direct photo-transformation half-life [days] of spirotetramat based on the quantum yield.
Season 30 ° lat. 40 ° lat. 50 ° lat. 60 ° lat. Spring 0.53 0.56 0.62 0.73
Summer 0.48 0.48 0.49 0.52
Fall 0.73 0.92 1.32 2.41
Winter 0.96 1.44 2.80 8.71

### – 168 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

In contrast to the GC-SOLAR model, the arithmetic m odel developed by Frank & Klöpffer 21 

### takes the conditions of cloudiness in Central Europe into con sideration. The environmental half-life of spirotet ramat, calculated according to Frank & Klöppfer modelling using the quantum yield of Φ = 0.00571 and the molar extinction coefficients from 292.5 to 490 nm, were spread from 0.5 to 52 days depending on the month.

Table B.8.4.2-6: Frank & Klöpffer modelling: Enviro nmental direct photo-transformation half-life [days] of spirotetramat in Central Europe based on the quantum yield.

Month Minimum Mean Maximum January 3.2 6.6 30 February 1.6 3.3 14
March 0.90 1.7 7.1 April 0.58 1.0 4.1 May 0.51 0.82 3.3
June 0.49 0.73 2.9 July 0.55 0.82 2.7 August 0.56 0.83 2.8 September 0.84 1.4 5.3
October 1.3 2.5 12
November 2.4 5.6 28
December 4.7 10 52



### Conclusion: A mean quantum yield of Φ = 0.00571 was calculated for spirotetramat. Based on the different arithmetic models (GC-SOLAR and Frank & Klöpffer) by means of resulting quantum yield and adsorption an environmental direct photo-transformation half-life of spirotetramat in a range of approx. 0.5 to appr ox. 1 day (mean) can be assessed for the periods of main use (late spring to summer). Direct phototransformation in water significantly contributes to the overall elim ination of spirotetramat in the environment.

Reviewer comments:

• Dark hydrolysis rate of spirotetramat is not menti oned (not measured) in the report but is essential to calculate net-photolysis rate. However, on request, the notifier gave the information that the Milli-Q water used had a pH of 6.2 – 6.4. Based on this pH value (dissolved spirotetramat is not expected to significantly shift the pH value) the hydrolysis DT 50 

### of spirotetramat is approx. 15 days. Considering an exposure period of 240 min (0.17 days) no signif icant degradation of spirotetramat owing to hydrolysis in the dark is expected. In this respect , the study results (assuming a zero dark rate) are
considered valid.

Reference: BYI08330-Enol: Determination of the quantum yield a nd assessment of the environmental half-life of the direct photodegradat ion in water Author(s), year: Heinemann, O., 2005b Study/report number: M-243787-01-2, MEF-04/438 Guideline(s): Phototransformation of Chemicals in W ater, part A: Direct Phototransformation, UBA, Berlin, Germany (1992-12); Test Method: ECETOC (Polychromatic Light Source), US-EPA Guideline No. 161-2 SUPP
GLP: Yes
Deviations: None Validity: Study considered acceptable

21 Frank, R., Klöpffer, W.: UBA Research Report No. 1060 2046 (1985)

### – 169 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Material and methods: Light source: Mercury immersion lamp TQ 150 (cut-of f filter < 295 nm) Actinometer solution: Uranyl oxalate (0.01 mol L -1 

### uranyl ions, 0.05 mole L -1 

### oxalic acid) Exposure of actinometer solution: 10 min at 25 °C Test solution: Spirotetramat-enol (unlabelled), 5.0 3 mg L -1 

### in Milli-Q-water (unbuffered) Exposure of test solution: 8.3 hrs Range of wave-lengths investigated: 295 – 490 nm Analysis of parent degradation: HPLC-UV UV/VIS-spectra of test substance: In test solution and in buffered solutions at pH 4, 7 and 9 (all 0.0 1 M, 10.3 mg test item L -1 

### ) Modelling of environmental half-life: a) GC-SOLAR b) Frank & Klöpffer

Findings: Based on actinometer (uranyl oxalate) degradation, a radiation intensity of approx. 7.25 × 10 16 

### photons (sec × 3 ml) -1 

### was calculated for the test unit. The UV/VIS absor ption spectrum of 10.3 mg L -1 

### spirotetramatenol in buffered solutions at pH 7 and pH 9 showed comparable absorption properties with a distinct maximum about 270 nm. In contrast, the UV/VIS absor ption spectrum in buffered solution at pH 4 showed no such maximum. The degradation of spirotetramat-enol under test conditions accounted for approx. 20 % over an irradiation period of 8.3 hrs. Mean degrada tion half-life under the experimental conditions (unbuffered solution in water) was calculated to be 1.39 days (SFO kinetics). Based on the degradation and the adsorption data, this resulted in a mean quantu m yield of direct photo-transformation in aqueous
solutions of Φ = 0.000252.

The environmental half-life of spirotetramat-enol, calculated according to GC-SOLAR using the quantum yield of Φ = 0.000252 and the molar extinction coefficients f rom 297.5 to 490 nm were spread from 8.6 – 173 days depending on the latitude and the season.

Table B.8.4.2-7: GC-SOLAR modelling: Environmental direct photo-transformation half-life [days] of spirotetramat-enol based on the quantum yield.
Season 30 ° lat. 40 ° lat. 50 ° lat. 60 ° lat. Spring 9.6 10 12 14
Summer 8.6 8.7 9.0 9.6
Fall 13 17 25 47
Winter 18 28 55 173



### In contrast to the GC-SOLAR model, the arithmetic m odel developed by Frank & Klöpffer 4 

### takes the conditions of cloudiness in Central Europe into con sideration. The environmental half-life of spirotet ramat, calculated according to Frank & Klöppfer modelling using the quantum yield of Φ = 0.000252 and the molar extinction coefficients from 292.5 to 490 nm, were spread from 9.1 to 1100 days depending on the month

.

### – 170 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.4.2-8: Frank & Klöpffer modelling: Enviro nmental direct photo-transformation half-life [days] of spirotetramat-enol in Central Europe base d on the quantum yield.

Month Minimum Mean Maximum January 65 140 620 February 31 65 280
March 17 33 140 April 11 20 78 May 9.5 15 61
June 9.1 14 54 July 10 15 51 August 10 16 52 September 16 27 100
October 26 50 230
November 49 110 570
December 98 220 1100



### Conclusion: A mean quantum yield of Φ = 0.000252 was calculated for spirotetramat-enol, the major metabolite of the parent spirotetramat. Based on the different arithm etic models (GC-SOLAR and Frank & Klöpffer) by mean s of resulting quantum yield and adsorption an enviro nmental direct photo-transformation half-life of spirotetramat in a range of approx. 9 to approx. 16 days (mean values) can be assessed for the periods of main use (late spring to summer). Direct phototrans formation in water is likely to contribute to the o verall elimination of spirotetramat-enol in the environmen t.

Reviewer comments:

• No dark control samples were included in the test design to account for hydrolysis processes. However, owing to the high stability of spirotetram at-enol in sterile buffer solutions at pH values fr om 4 to 9 (Heinemann, 2004b), the study is considered valid (no hydrolysis assumed).

### – 171 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.4.3 Ready biodegradability of the active substa nce (OECD Annex IIA, 7.7)

Reference: BYI 08330 Biodegradation Author(s), year: Weyers, A., 2005 Study/Report number: M-263287-01-1, 2005/0077/01 Guideline(s): ECCD 92/69/EEC C.4-D, OECD 301 F
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substance: Spirotetramat (unlabelled), purity 97.4, batch MIX-Batch 08045/0014
Reference substance: Na-benzoate Inoculum: Aeration tank of a waste water plant trea ting predominately domestic sewage (30 mg L -1 

### )
Treatments: • Blank control

• Reference substance: Na-benzoate (100 mg L -1 

### )

• Test substance: Spirotetramat (100 mg L -1 

### )

• Toxicity control: Spirotetramat (100 mg L -1 

### ) and Na-benzoate (100 mg L -1 

### ) pH of the test vessel at the end of test: 7.3 Analysis: Biochemical oxygen demand (corrected for oxygen consumption by nitrification) Incubation conditions: 22 ± 2 °C, 28 days

Findings:

Table B.8.4.3-1: Biodegradation of spirotetramat an d reference compound [% of theoretically possible degradation]. DAT Reference substance Test substance Toxicity con trol

4 73 0 29
12 88 1 37
20 91 0 38
28 91 1 40



### Within 28 days almost no degradation (maximum 1 %) was determined for spirotetramat. The reference substance (Na-benzoate) has reached level for ready biodegradability by 14 days.

Conclusion: Spirotetramat is considered to be not readily biode gradable.

Reviewer comments:
None

### – 172 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.4.4 Degradation in aquatic systems (OECD Annex IIA, 7.8)

B.8.4.4.1 Aerobic biodegradation in aquatic systems (OECD Annex IIA, 7.8.1)

This point is covered by section B.8.4.4.3.

B.8.4.4.2 Anaerobic biodegradation in aquatic syste ms (OECD Annex IIA, 7.8.2)

This point is covered by section B.8.4.4.3.

B.8.4.4.3 Water/sediment studies (OECD Annex IIA, 7 .8.3)

Two water sediment studies were conducted:

• Aerobic water/sediment study with two test systems , [3- 14 

### C] and [5- 14 

### C] label

• Anaerobic water/sediment study with one test syste m, [3- 14 

### C] label

Reference: BYI08330: Aerobic Aquatic Metabolism Author(s), year: Menke, U., 2006c Study/report number: M-269307-01-2, MEF-04/511 Guideline(s): 95/36/EC, OECD Guideline 308 (2002), US-EPA N 162-4 (1982)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods:
Test substances: • [Azaspirodecenyl-3- 14 

### C]-spirotetramat, 3.67 MBq mg -1



### , > 98 % radiochemical purity (HPLC), batch BECH 0955

• [Azaspirodecenyl-5- 14 

### C]-spirotetramat, 4.03 MBq mg -1



### ,

> 

### 98 % radiochemical purity (HPLC), batch BECH 0956 Labels regarded as replicates Reference substances: Spirotetramat (unlabelled), s pirotetramat-enol, spirotetramat-ketohydroxy, spirotetramat-MA-amide, spirotetramat-di-hydroxy, s pirotetramat-oxo-enol,
benzoic acid Application rate: 29 μg L -1 

### (under the assumption of 288 g ai ha -1 

### , 100 cm water depth) Co-solvent: Acetonitrile (0.015 %, v/v) Incubation set-up: 102.4 g (‘Hönniger Weiher’) and 182.1 g (‘Anglerweiher’) dry sediment (2 mm sieved), 519 mL of water (sediment/water, 1/3, w/v) , equivalent to 2 cm sediment and 6 cm water column, water phase kept in motion b y stirring, sediment stagnant Acclimatization period: Approx. 7 days Test duration: 120 days
Incubation conditions: 20 ± 1 °C in darkness Volatile traps: Polyurethane foam (organic volatile s), soda lime (CO 2 

### ), identity of 14 

### CO 2 

### confirmed by Grignard Reaction Monitoring: pH, O 2 

### , redox potential Analysis: Water phase radio-assayed by LSC and concentrated using a speedvac

concentrator.

### – 173 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Sediment phase extracted 3 times with 100 ml of acetonitril e/water/formic acid (50/50/0.5, v/v/v). All three extracts were combine d ( ‘organic extract’ ). Then the
sediment was extracted 2 times with 100 ml of aceto nitrile/1 M HCl (50/50, v/v) and once with 100 ml of pure acetonitrile (all thre e combined to ‘acid extract’ ). Organic extracts were separately concentrated using a speedvac concentrator. Aliquots of the acidic extracted soils from 120 DAT samples were separated into humic acids, fulvic acids and humins.
Non extractable residues were radio-assayed by combustion LSC. LOQ(HPLC) approx. 0.5 – 0.8 % of AR Analytical techniques: LSC, HPLC-UV/RAD, TLC, LC-MS /MS (ESI) Kinetic evaluation: Simple first order (SFO) kineti cs, MatLab

Table B.8.4.4.3-1: Physicochemical characteristics of the water/sediment matrices.

Name ‘Hönniger Weiher’ ‘Anglerweiher’

Geographic location Wipperfürth,NorthrhineWestfalia, Germany Leverkusen,

NorthrhineWestfalia, Germany

Type of aquatic system Meso- /oligotrophic Oligotrophic

Texture (USDA) Loam Loamy sand Sand (USDA) [%] 32.9 81.0 Silt (USDA) [%] 49.8 11.3 Clay (USDA) [%] 17.3 7.7 pH (0.01 M CaCl 2 ) 5.6 6.8 pH (water) 6.1 7.3 Organic C [%] 4.42 0.99 Total N [%] 0.35 0.09 CEC [meq Ba 2+ kg -1 ] 165 61 Carbonate [%] 0.3 < 0.1 Total P [mg kg -1 ] 770 224 Redoxpotential [mV] -159 -54 Water content [%] 58.1 33.7 Sediment

Microbial activity [mg CO 2 h -1 kg -1 ] – sampling / start / 120 DAT 36 / 43 / 25 10 / 10 / 8

pH (sampling) 6.68 7.16 Total hardness [°dH] 4.3 10.2 Oxygen content [mg L -1 ] – sampling / 120 DAT 3.08 / 7.23 6.60 / 7.77 Redoxpotential [mV] – sampling / 120 DAT 455 / 378 4 61 / 366 DOC [mg L -1 ] 4 < 2 Total N [mg L -1 ] 5.2 5.6 Water

Total P [mg L -1 ] 0.11 < 0.03



### Findings: The oxygen content of the water phase of the sample s was in a range of 7.2 – 8.5 mg L -1 

### (‘Hönniger Weiher’) and 7.7 – 8.3 mg L -1 

### (‘Anglerweiher’), indicating aerobic conditions th roughout both experiments. The pH in the water phase was slightly increasing from 6.8 by 0 DAT to approx. 7.5 by 120 DAT (‘Hönniger Weiher’ ) and from 7.9 – 8.2 (‘Anglerweiher’). In the sedimen t the pH stayed more or less unchanged with values about 6.8 (‘Hönniger Weiher’) and 7.3 (‘Anglerweiher’). T he redox potential in the water phase was fairly co nstant in a range of +353 – +428 mV (‘Hönniger Weiher’) an d +351 – +397 mV (‘Anglerweiher’), in the sediment the redox potential was in a range of +42 – +189 mV (‘H önniger Weiher’) and +154 – +360 mV (‘Anglerweiher’ ) with a steady decrease in the initial time of the l atter test system. In summery, in both systems wate r and sediment stayed aerobically throughout the study pe riod.

### – 174 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Total mass balance was in a range of 94.4 – 102.6 % of AR for both systems and both labels. Formation of

14 

### CO 2 

### using 3- 14 

### C accounted for maximum of 11.0 % (‘Hönniger Weiher ) and 24.0 % (‘Anglerweiher’) of AR at study termination. Formation of 14 

### CO 2 

### from 5- 14 

### C labelled spirotetramat was approx. half indicatin g that this part of the molecule is less available to biod egradation than the 3- 14 

### C position. Formation of NER steadily increased in both systems towards study te rmination with a maximum of 40.7 % of AR by 91 DAT.

Spirotetramat quickly degraded in both water/sedime nt systems, by 7 DAT no spirotetramat was detectabl e in the water or in the sediment phase. Transfer of the test item into the sediment phase was almost ne gligible with a maximum occurrence of 3.4 % of AR by 1 DAT. No distinct differences between labels used were observed. Degradation of spirotetramat led to extin ct formation of the common hydrolysis product spirotetramat-enol (maximum 99.0 % of AR by 14 DAT in the entire system of ‘Hönniger Weiher’ and 92.1 % of AR by 7 DAT in ‘Anglerweiher’). Maximum amounts of spirotetramat-enol found in the sediment were 41.2 % and 15.7 % of AR (‘Hönniger Weiher’ and ‘Ang lerweiher’). After reaching maximum, amounts of spirotetramat-enol significantly decreased thereaft er in both, water and sediment. As a second major metabolite spirotetramat-ketohydroxy was formed > 1 0 % of AR in the entire system with a more or less increasing trend towards study termination (maximum in water, sediment, entire system: 17.4 %, 42.8 % and 50.8 % of AR, respectively). Three further identifi ed metabolites (spirotetramat-MA-amide, spirotetram at-oxoenol isomer and spirotetramat-di-hydroxy) did not e xceed 9.2 %, 6.0 % and 9.8 % of AR, respectively (a ll significantly decreasing thereafter). Unidentified radioactivity (sum) was below 8.1 % of AR in the en tire system of both test systems.

Table B.8.4.4.3-2: Distribution and recovery of rad ioactivity [% of AR] after application of [3- 14 

### C] or [5-

14 

### C] labelled spirotetramat (288 g ai ha -1 

### ) to the aerobic water/sediment system ‘Hönniger We iher’.

Sediment Label DAT 14 CO 2 Organic volatiles Water Organic
extract Acid

extract Total NER Total

0 na na 98.8 2.3 0.4 2.7 0.2 101.6
0.2 < 0.1 < 0.1 92.5 4.6 0.8 5.4 0.2 98.2

1 < 0.1 < 0.1 86.4 10.4 1.6 12.0 0.8 99.2
3 < 0.1 < 0.1 83.5 13.9 1.9 15.8 0.8 100.2

7 0.1 < 0.1 78.6 18.2 2.7 20.9 0.7 100.3

14 0.2 < 0.1 69.2 24.4 3.0 27.4 1.2 97.8
30 2.1 < 0.1 26.9 17.3 20.9 38.2 30.0 97.3
60 3.7 < 0.1 29.2 17.3 17.7 35.0 29.4 97.3
91 8.0 < 0.1 22.7 15.7 17.8 33.5 32.0 96.2 3- 14 C

120 11.0 < 0.1 17.8 14.9 19.9 34.8 32.9 96.4

0 na na 94.8 1.5 0.1 1.6 < 0.1 96.4
0.2 < 0.1 < 0.1 94.4 5.2 0.8 6.0 0.2 100.6

1 < 0.1 < 0.1 87.7 9.6 1.4 11.0 0.5 99.2
3 < 0.1 < 0.1 84.0 12.3 1.8 14.1 0.6 98.7

7 0.1 < 0.1 76.0 19.1 3.1 22.2 1.1 99.5

14 0.1 < 0.1 70.7 26.7 3.2 29.9 0.8 101.5
30 0.8 < 0.1 24.3 19.7 21.7 41.4 31.7 98.1
60 2.2 < 0.1 12.9 18.6 31.4 50.0 36.7 101.8
91 4.2 < 0.1 7.6 14.0 29.4 43.4 40.7 95.9 5- 14 C

120 5.9 < 0.1 8.2 13.8 30.2 44.0 36.3 94.4

### – 175 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.4.4.3-3: Distribution and recovery of rad ioactivity [% of AR] after application of [3- 14 

### C] or [5-

14 

### C] labelled spirotetramat (288 g ai ha -1 

### ) to the aerobic water/sediment system ‘Anglerweihe r’.

Sediment Label DAT 14 CO 2 Organic volatiles Water Organic
extract Acid

extract Total NER Total

0 na na 95.5 1.4 0.2 1.6 < 0.1 97.1
0.2 < 0.1 < 0.1 93.3 4.2 0.4 4.6 0.1 97.9

1 < 0.1 < 0.1 90.4 7.9 1.3 9.2 1.0 100.7
3 0.1 < 0.1 88.2 8.6 2.0 10.6 2.0 100.9

7 0.1 < 0.1 83.3 12.4 2.5 14.9 3.8 102.2

14 0.4 < 0.1 79.2 14.0 4.0 18.0 5.0 102.6
30 3.0 < 0.1 49.3 11.4 15.5 26.9 22.1 101.3
60 10.2 < 0.1 34.2 9.2 15.0 24.2 29.3 98.0
91 19.7 < 0.1 27.3 6.5 14.9 21.4 30.4 98.8 3- 14 C

120 24.0 < 0.1 20.2 7.4 15.7 23.1 32.3 99.6

0 na na 97.5 1.8 0.2 2.0 < 0.1 99.5
0.2 < 0.1 < 0.1 94.8 4.3 0.4 4.7 0.2 99.6

1 < 0.1 < 0.1 89.4 8.1 1.3 9.4 1.2 100.0
3 0.1 < 0.1 87.3 8.2 1.9 10.1 2.2 99.6

7 0.1 < 0.1 80.9 13.0 2.8 15.8 3.6 100.4

14 0.2 < 0.1 77.3 14.6 3.5 18.1 4.8 100.3
30 1.2 < 0.1 54.6 13.7 14.0 27.7 18.4 98.9
60 3.7 < 0.1 31.2 15.9 17.3 33.2 30.5 98.7
91 8.2 < 0.1 24.0 14.4 18.8 33.2 33.2 98.6 5- 14 C

120 13.5 < 0.1 17.3 13.2 18.8 32.0 33.9 96.7– 176 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.4.4.3-4: Characterization of extractableradioactivity [% of AR] after application of [3-14C] or [5-14C] labelled spirotetramat (288 g ai ha-1) to theaerobic water/sediment system ‘Hönniger Weiher’.Spirotetramat Spirotetramat-enol Spirotetramat-ketohydroxy Spirotetramat-MA-amide Unidentified RALabelDATWater Sed.EntiresystemWater Sed.EntiresystemWater Sed.EntiresystemWater Sed.EntiresystemWater Sed.Entiresystem082.4 2.2 84.615.7 < LOQ15.7 < LOQ < LOQ < LOQ < LOQ < LOQ < LOQ 0.7 0.5 1.20.270.2 3.1 73.321.4 1.522.9 < LOQ < LOQ < LOQ < LOQ < LOQ < LOQ 0.9 0.8 1.7144.63.448.040.4 6.947.3 0.9 < LOQ 0.9 < LOQ < LOQ < LOQ 0.5 1.7 2.236.3 1.9 8.262.913.476.313.5 < LOQ13.5 < LOQ < LOQ < LOQ 0.9 0.6 1.57< LOQ < LOQ < LOQ77.420.597.9< LOQ < LOQ < LOQ < LOQ < LOQ < LOQ 1.1 0.4 1.514< LOQ < LOQ < LOQ68.227.295.4 < LOQ < LOQ < LOQ < LOQ < LOQ < LOQ 0.9 0.1 1.030< LOQ < LOQ < LOQ23.334.157.4 3.1 2.8 5.9 < LOQ < LOQ < LOQ 0.4 1.3 1.760< LOQ < LOQ < LOQ19.832.051.8 3.3 2.2 5.52.1< LOQ 2.14.10.7 4.891< LOQ < LOQ < LOQ13.415.528.9 3.312.916.2 2.01.0 3.04.04.1 8.1[3-14C]120< LOQ < LOQ < LOQ < LOQ21.021.017.412.830.2< LOQ < LOQ < LOQ 0.3 1.0 1.3077.0 1.5 78.517.5 < LOQ17.5 < LOQ < LOQ < LOQ < LOQ < LOQ < LOQ 0.4 0.2 0.60.273.33.376.620.0 1.721.7 < LOQ < LOQ < LOQ < LOQ < LOQ < LOQ 1.0 0.9 1.9141.4 2.9 44.344.9 6.651.5 0.7 < LOQ 0.7 < LOQ < LOQ < LOQ 0.6 1.5 2.136.5 1.6 8.161.312.173.413.5< LOQ13.5 < LOQ < LOQ < LOQ2.70.43.17< LOQ < LOQ < LOQ75.321.897.1 < LOQ < LOQ < LOQ < LOQ < LOQ < LOQ 0.8 0.5 1.314< LOQ < LOQ < LOQ69.229.899.01.0 < LOQ 1.0 < LOQ < LOQ < LOQ 0.5 0.1 0.630< LOQ < LOQ < LOQ19.736.456.1 3.7 3.5 7.2 < LOQ < LOQ < LOQ 0.91.62.560< LOQ < LOQ < LOQ 6.441.247.6 6.0 7.713.7 < LOQ < LOQ < LOQ 0.5 1.1 1.691< LOQ < LOQ < LOQ < LOQ29.029.0 3.913.617.51.4< LOQ1.42.3 0.83.1[5-14C]120< LOQ < LOQ < LOQ < LOQ < LOQ < LOQ 8.042.850.8< LOQ < LOQ < LOQ 0.2 1.2 1.4LOQ denotes limit of quantification (i.e. approx. 0.5 – 0.8 % of AR)– 177 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.4.4.3-5: Characterization of extractableradioactivity [% of AR] after application of [3-14C] or [5-14C] labelled spirotetramat (288 g ai ha-1) to theaerobic water/sediment system ‘Anglerweiher’.SpirotetramatSpirotetramat-enolSpirotetramat-ketohydroxySpirotetramat-MA-amideSpirotetramat-oxo-enol isomerSpirotetramat-di-hydroxyUnidentified RALabelDATWat. Sed. Total Wat. Sed. Total Wat. Sed. Total Wat. Sed. Total Wat. Sed. Total Wat. Sed. Total Wat. Sed. Total070.2 1.4 71.624.3 < LQ24.3 < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ <LQ < LQ < LQ 1.0 0.2 1.20.260.72.863.532.0 1.233.2 < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ <LQ < LQ < LQ 0.6 0.5 1.1138.9 2.0 40.950.7 5.956.6 0.8 < LQ 0.8 < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ 0.1 1.4 1.535.8 0.6 6.478.7 7.386.0 2.4 0.5 2.9 < LQ < LQ < LQ < LQ < LQ < LQ < LQ< LQ < LQ 1.2 2.2 3.47< LQ < LQ < LQ79.711.391.03.4 1.0 4.4 < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ< LQ 0.3 2.7 3.014< LQ < LQ < LQ72.515.788.2 2.6 1.8 4.4 1.6 < LQ 1.6 < LQ < LQ < LQ < LQ <LQ < LQ 2.5 0.5 3.030< LQ < LQ < LQ39.913.453.3 6.211.918.1 2.4 < LQ 2.4 < LQ < LQ < LQ < LQ < LQ < LQ 0.71.5 2.260< LQ < LQ < LQ15.1 2.217.36.718.825.5 7.8 1.4 9.2< LQ < LQ < LQ < LQ 1.2 1.2 4.7 0.7 5.491< LQ < LQ < LQ 6.3 < LQ 6.3 1.813.815.6 6.3 < LQ 6.3 3.7 < LQ 3.74.3 5.5 9.8 4.92.06.9[3-14C]120< LQ < LQ < LQ 8.2 6.414.6 5.611.316.9 < LQ < LQ < LQ6.0< LQ6.0< LQ < LQ < LQ 0.43.13.5071.2 1.7 72.925.7 < LQ25.7 < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ <LQ < LQ < LQ 0.6 0.2 0.80.264.92.167.029.2 2.131.3 < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ <LQ < LQ < LQ 0.7 0.4 1.1139.9 2.0 41.946.0 5.451.4 2.0 0.5 2.5 < LQ < LQ < LQ < LQ < LQ < LQ < LQ< LQ < LQ 1.5 1.5 3.035.4 0.6 678.67.185.7 2.0 0.5 2.5 < LQ < LQ < LQ < LQ < LQ < LQ < LQ< LQ < LQ 1.2 1.9 3.17< LQ < LQ < LQ77.914.292.12.2 < LQ 2.2 < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ < LQ 0.8 1.6 2.414< LQ < LQ < LQ71.514.686.1 3.1 2.9 6.0 1.6 < LQ 1.6 < LQ < LQ < LQ < LQ <LQ < LQ 1.0 0.6 1.630< LQ < LQ < LQ39.715.555.26.78.615.3 2.1 < LQ 2.1 < LQ < LQ < LQ < LQ < LQ < LQ 3.13.66.760< LQ < LQ < LQ15.0 4.119.1 5.723.128.86.1 2.1 8.2< LQ < LQ < LQ4.2 2.5 6.70.2 1.5 1.791< LQ < LQ < LQ 7.9 4.712.6 2.123.926.0 3.6 1.9 5.53.8< LQ3.8< LQ 2.1 2.16.60.67.2[5-14C]120< LQ < LQ < LQ 8.0 < LQ 8.0 2.431.433.82.0 < LQ 2.0 < LQ2.32.3 < LQ < LQ < LQ 4.8 0.5 5.3LQ denotes limit of quantification (i.e. approx. 0.5 – 0.8 % of AR)

### – 178 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.4.4.3-6: Calculated half-lives [days] of spirotetramat in the aerobic water/sediment systems ‘Hönniger Weiher’ and ‘Anglerweiher’ based on SFO k inetics. Water Total system Dissipation Degradation System Label DT 50 [days] DT 90

[days] R 2 DT 50

[days] DT 90

[days] R 2 Kinetics

‘Hönniger Weiher’ Labels used as replicates 1.00 3.31 0.99 1.06 3.52 1.00 SFO ‘Anglerweiher’ Labels used as replicates 1.02 3.40 0.99 1.05 3.50 0.99 SFO



### Conclusion: Spirotetramat quickly degraded in two contrasting a erobic water/sediment systems with a degradation ha lflife in the entire system of 1.0 day (both systems, SFO, R 2 

### ≥ 0.99). Dissipation from the water layer was fast
as well. No distinct differences between labels use d were observed. As major metabolites spirotetramat -enol (maximum in the total system 99.0 % of AR by 14 DAT , decreasing thereafter) and spirotetramatketohydroxy (maximum in the total system 50.8 % of AR at study termination) were observed. Other identified substances (spirotetramat-MA-amide, spir otetramat-oxo-enol isomer and spirotetramat-di-hydr oxy) did not exceed 9.8 % of AR in the total system indi vidually, non identified radioactivity accounted fo r maximum of 8.1 % of AR in sum. Formation of 14 

### CO 2 

### (maximum 24.0 % of AR in one system by study termination) indicate significant mineralization of the test item, NER accounted for maximum 40.7 % of AR by
91 DAT.

Reviewer comments:

• Dissipation and degradation rates of spirotetramat , spirotetramat-enol and spirotetramat-ketohydroxy were (re-)calculated by the notifier (supplemental studies, Röpke, 2006b).

Reference: BYI08330[azaspirodecenyl-3- 14 

### C]: Anaerobic aquatic metabolism Author(s), year: Shepherd, J., Arthur, E. L., Domin ic, A., 2005 Study/report number: M-261943-01-1, MEFNX015 Guideline(s): US-EPA N 162-3 (1982, 1989), OECD Gui deline 308 (2002), Canada PMRA DACO Number 8.2.3.5.6 (1987)
GLP: Yes
Deviations: None Validity: Study considered acceptable

Material and methods: Test substances: [Azaspirodecenyl-3- 14 

### C]-spirotetramat, 3.67 MBq mg -1 

### , > 99 % radiochemical purity (HPLC), batch BECH 0936 Reference substances: Spirotetramat, spirotetramat- enol (both 3- 14 

### C labelled) Application rate: 14 μg L -1 

### (assuming 288 g ai ha -1 

### and 200 cm water depth), 10fold concentration
used for metabolite identification work Co-solvent: Acetonitrile (< 0.1 %, v/v) Incubation set-up: 50 g (dry weight) sediment (2 mm sieved), 150 mL of pond water, sediment/water ratio 1/3 (w/w) Acclimatization period: 27 days Test duration: 120 days
Incubation conditions: 20 ± 1 °C in darkness with a N 2 

### -atmosphere Volatile traps: 2 × 2.0 M KOH (30 ml), ethylene gly col (30 ml), 1 M sulphuric acid (30 ml) Monitoring: pH, O 2 

### , redox potential Analysis: The water phase was acidified with formic acid, radio-assaye d by LSC and concentrated by rotary evaporation.

### – 179 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

The sediment phase was extracted once with 100 ml of acetonitri le/water/formic acid (90/10/0.1, v/v/v) following ASE extraction at 80 °C using the same solvent. The two extracts were combined, acidified and radio -assayed by LSC. An aliquot was concentrated by rotary evaporation and analysed by HPLC.
Non extractable residues were radio-assayed by combustion LSC. Analytical techniques: LSC, HPLC-RAD, LC-ESI/MS Kinetic evaluation: Simple first order (SFO) kineti cs, GraphPad PRISM

Table B.8.4.4.3-7: Physicochemical characteristics of the water/sediment matrix.

Name ‘Lawrence’

Geographic location Nelson Environmental Study Area, Lawrence, KS, Jefferson County, USA Texture (USDA) Clay loam Sand (USDA) [%] 21 Silt (USDA) [%] 40 Clay (USDA) [%] 39 pH (0.01 M CaCl 2 ) 7.1 pH (water) 7.6 pH (saturated paste) 7.4 Redox potential [mV] – start / 120 DAT 68.5 / -38.5 Organic C [%] 0.7 CEC [meq kg -1 ] 327 Field MC at 0.33 bar [%] 35.8 Bulk density [g cm -3 ] 1.1 Sediment

Microbial biomass [10 8 cells g -1 ] – start / 120 DAT 2.00 / 1.07

pH 7.5 Total hardness (CaCO 3 ) [mg L -1 ] 139 Oxygen content [mg L -1 ] – start / 120 DAT 0.0 / 0.0 Redox potential [mV] – start / 120 DAT 55.2 / -20.0 DOC [mg L -1 ] – start / 120 DAT 8.4 / 9.3 Water

Microbial biomass [10 6 cells g -1 ] – start / 120 DAT 3.48 / 2.55



### Findings: Throughout the study, the average redox potential ( +25 mV in water, +15 mV in sediment) and the averag e dissolved oxygen concentration ( ≤ 0.1 mg L -1 

### ) remained low indicating that an anaerobic reducin g environment had been maintained. The average pH of the water ranged from 6.7 to 7.3.

The mean material balance for the study ranged from 90.1 to 102.2 % of AR.

The concentration of spirotetramat in water decreas ed from 81.3 % of AR by 0 DAT to 57.2 % of AR by 2 DAT and continued to decrease to 0.0 % of AR by 14 DAT. In the sediment the concentration of spirotetramat decreased from 8.8 % of AR by 0 DAT t o 0.0 % of AR by 14 DAT. The only transformation product detected in water was spirotetramat-enol wi th a maximum concentration of 80.5 % of AR by 14 DA T, hardly decreasing thereafter to a final amount of 7 3.3 % of AR. In the sediment spirotetramat-enol was detected with a maximum amount of 28.5 % of AR by 2 1 DAT without significant changes towards the furth er incubation. No other metabolites were observed in t he water or in the sediment phase.

### – 180 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.4.4.3-8: Distribution and recovery of rad ioactivity [% of AR] after application of [3- 14 

### C] labelled spirotetramat (288 g ai ha -1 

### ) to the anaerobic water/sediment system ‘Lawrence’ (mean of duplicate samples).

Label DAT 14 CO 2 Organic volatiles Water Sediment NER Total

0 0.0 0.0 81.3 8.8 0.0 90.1
2 0.6 0.0 79.8 17.7 0.0 98.1

7 0.5 0.0 76.6 21.2 0.0 98.3
9 0.6 0.0 77.2 22.3 0.0 100.1

14 0.6 0.0 80.2 21.4 0.0 102.2
21 0.6 0.0 75.7 25.5 0.0 101.8
28 0.7 0.0 76.4 24.1 0.0 101.3
63 0.7 0.0 72.3 28.0 0.0 101.0
90 0.7 0.0 76.4 24.0 0.5 101.5 [3- 14 C]

120 0.8 0.0 73.3 26.7 0.6 101.4



### Table B.8.4.4.3-9: Characterization of extractable radioactivity [% of AR] after application of [3- 14 

### C] labelled spirotetramat (288 g ai ha -1 

### ) to the anaerobic water/sediment system ‘Lawrence’

. Spirotetramat Spirotetramat-enol Label DAT Water Sediment Total Water Sediment Total

0 83.3 9.3 92.6 0.0 0.0 0.0
0 79.3 8.2 87.5 0.0 0.0 0.0
2 53.1 7.2 60.3 26.6 10.2 36.8
2 61.4 7.1 68.5 18.5 10.9 29.4

7 11.0 2.2 13.2 64.2 19.1 83.3

7 8.8 0.0 8.8 69.2 21.1 90.3
9 9.1 2.2 11.3 67.9 19.3 87.2
9 6.4 0.0 6.4 70.8 23.1 93.9

14 0.0 0.0 0.0 80.5 20.9 101.4

14 0.0 0.0 0.0 79.8 21.1 100.9
21 0.0 0.0 0.0 78.2 22.6 100.8
21 0.0 0.0 0.0 73.1 28.5 101.6
28 0.0 0.0 0.0 77.0 23.1 100.1
28 0.0 0.0 0.0 75.9 25.0 100.9
63 0.0 0.0 0.0 72.5 27.8 100.3
63 0.0 0.0 0.0 72.1 28.2 100.3
90 0.0 0.0 0.0 76.2 24.0 100.2
90 0.0 0.0 0.0 76.5 23.9 100.4
120 0.0 0.0 0.0 74.9 25.0 99.9 [3- 14 C]

120 0.0 0.0 0.0 71.7 28.4 100.1



### Table B.8.4.4.3-10: Calculated half-lives of [3- 14 

### C] [days] labelled spirotetramat in the anaerobic water/sediment system ‘Lawrence’ based on SFO kinet ics. Water Total system Dissipation Degradation System DT 50

[days] DT 90

[days] R 2 DT 50 [days] DT 90

[days] R 2 Kinetics

‘Lawrence’ 2.8 9.2 0.99 2.8 9.3 0.99 SFO



### Conclusion: Under anaerobic water/sediment conditions spirotetr amat was completely transformed into its major metabolite spirotetramat-enol which can be consider ed stable under the experimental conditions. Spirotetramat-enol reached maximum amounts in the w ater of 80.5 % of AR by 14 DAT and in the sediment

### – 181 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

of 28.5 % of AR by 21 DAT hardly changing thereafte r. The degradation half-life of spirotetramat was calculated to be 2.8 days (R 2 

### = 0.99) in the total system, respective DT 90 

### was 9.3 days.

Reviewer comments:
None

B.8.4.4.4 Supplemental studies

Reference: Kinetic Evaluation of the Aerobic Aquatic metabolis m of BYI08330, BYI08330-enol and BYI08330-ketohydroxy in water sed iment systems Author(s), year: Röpke, B., 2006b Study/Report number: M-277415-01-1, MEF-06/279 Guideline(s): Recommendations of FOCUS working grou p on degradation kinetics (FOCUS, 2006) GLP: Not applicable Deviations: Not applicable Validity: Study considered acceptable

Material and methods: The behaviour of spirotetramat and its major water/ sediment metabolites spirotetramat-enol and spirotetramat-ketohydroxy in an aquatic environment was investigated by kinetic evaluation of the aero bic
water/sediment studies conducted with the two test systems ‘Hönniger Weiher’ and ‘Anglerweiher’.

Simple first order (SFO) dissipation half-life of s pirotetramat in the water phase is based on SFO kin etics. Additionally, overall SFO degradation of spirotetra mat and spirotetramat-enol and spirotetramat-ketohy droxy in the entire system were derived using a four-comp artment model (no additional pathways to sink).

Figure B.8.4.4.4-1: Multi-compartment model used fo r fitting residues of spirotetramat and

metabolites in water/sediment studies (entire syste m).

The goodness of fit was assessed by visual inspecti on, R 2



### , χ 2 

### error and t-test statistics. Calculations were performed with MatLab 7.0.4.365 (MatLab, 2005). Dif ferent equations were integrated by a Runge-Kutta method, and the Levenberg-Marquardt algorithm was u sed for non-linear parameter optimisation. Spirotetramat (SFO)

Spirotetramat-enol (SFO)

Spirotetramat-ketohydroxy (SFO)

Sink

### – 182 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

In case of spirotetramat-ketohydroxy, the 3 DAT sam pling of ‘Hönninger Weiher’ was considered an outli er, amounts of spirotetramat-ketohydroxy were set to < LOQ.

Findings:

Table B.8.4.4.4-1: Calculated dissipation (water ph ase) and degradation (entire system) half-life of spirotetramat and the metabolites spirotetramat-eno l and spirotetramat-ketohydroxy in two aerobic water/sediment systems based on SFO kinetics (both labels handled as replicates).

Substance Compartment Test system DT 50 [days] DT 90

[days] R 2 χ 2

error [%] Kinetics

‘Hönniger Weiher’ 0.82 2.73 0.99 6.9 SFO Water a (Dissipation) ‘Anglerweiher’ 0.74 2.45 0.97 12.3 SFO

‘Hönniger Weiher’ 0.86 2.85 0.99 6.2 SFO Spirotetramat Entire system

(Degradation) ‘Anglerweiher’ 0.70 2.34 0.97 11.9 SFO

‘Hönniger Weiher’ 59.0 196 0.93 13.4 P SFO  M SFO Spirotetramat- enol Entire system

(Degradation) ‘Anglerweiher’ 37.9 126 0.93 15.6 P SFO  M SFO

‘Hönniger Weiher’ Not evaluable, no decline P SFO  M1 SFO  M2 SFO Spirotetramat- ketohydroxy Entire system

(Degradation) ‘Anglerweiher’ Not evaluable, no decline P SFO  M1 SFO  M2 SFO

a Re-calculated by the reviewer (see comments)



### Conclusion: Degradation of spirotetramat in the entire system w as calculated to be 0.86 and 0.70 days for the ‘Hön niger Weiher’ and ‘Anglerweiher’ test system, respectivel y (following SFO kinetics, R 2 

### ≥ 0.97, χ 2 

### ≤ 6.9). Based on multi-compartment modelling, metabolite spirotetram at-enol degraded with a DT 50 

### in the range of 37.9 – 59.0 days in the total system (all SFO, R 2 

### > 0.93). Metabolite spirotetramat-ketohydroxy was considered to be stable in both systems.

Reviewer comments:

• The multi-compartment model used by the notifier d id not include supplemental pathways from the parent and the metabolite spirotetramat-enol to the sink. Additional calculations by the reviewer including these supplemental pathways did not impro ve the model. Owing to the extensive formation of spirotetramat-enol from the parent, no additiona l pathway (e.g. from parent to sink) has to be assumed. Therefore, the multi-compartment model use d by the notifier is considered appropriate.

• For both metabolites, additional calculations are presented for the dissipation in the water and
sediment based on the maximum occurrence as startin g point (following SFO kinetics). Since the aquatic risk assessment of both metabolites is sole ly based on the degradation half-life in the total system, these data are not considered relevant (not presented her).

• For calculating the dissipation of spirotetramat i n the water system, 0 DAT amounts of metabolites were not added to the measured 0 DAT amounts of spi rotetramat (as done for the entire system). Therefore, dissipation in the water phase was re-ca lculated by the reviewer following current guidance (SANCO/10058/2005, vers. 1.0, 2005).

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.4.4.5 Summary on fate & behaviour in aquatic sy stems

Aquatic hydrolysis

One hydrolysis study in dark sterile buffer solutio ns at pH 4, 7 and 9 at 20 °C (30 days), 25°C (up to 31 days) and 50 °C (up to 48 days) using [3- 14 

### C] and [5- 14 

### C] labelled spirotetramat was carried out. Spirotet ramat was shown to hydrolyse in strong dependence of the pH v alue and temperature, DegT 50 

### values at 25 °C were 32.5, 8.6 and 0.32 days at pH 4, 7 and 9, respectiv ely. Hydrolysis rates decreased with decreasing temperature. No significant differences between lab els used were observed. No volatiles were formed. During hydrolysis, spirotetramat almost exclusively degraded to spirotetramat-enol, which was shown to be stable under conditions of sterile hydrolysis in a separate hydrolysis study (at pH 4, 7 and 9, 25 and 50 °C, using [3- 14 

### C] and [5- 14 

### C] labelled spirotetramat-enol).

NH

O O O O O

H

NH

O O H O

H

Spirotetramat Spirotetramat-enol



### Figure B.8.4.4.5-1: Proposed hydrolysis pathway of spirotetramat in sterile buffer solutions.

Aquatic photolysis

Aquatic photolysis of spirotetramat was investigate d in sterile buffer solutions at pH 5.0 and in ster ilized natural water at pH 7.9 using [3- 14 

### C] and [5- 14 

### C] labelled parent. The test systems were continuou sly irradiated with a xenon arc lamp (> 290 nm) for 7 d ays and 10 days (sterile buffer solution and natura l water, respectively) to simulate the impact of natural lig ht. Both experiments were carried out at 25 °C.

Under irradiation in sterile buffer solutions at pH 5.0, spirotetramat degraded with a half-life of 2. 7 days (following SFO kinetics, R 2 

### = 0.98, net photolysis half-life corrected for hyd rolysis was 3.0 days). The experimental half-life (including photolysis and hy drolysis) corresponds to 12.0, 9.3 and 11.4 days un der environmental midday summer light in Athens (Greece , EU), Phoenix (AZ, USA) and Edmonton (Alberta, Canada). In the dark control, DegT 50 

### was 26.2 days (owing to hydrolysis, only) indicati ng that photodegradation of spirotetramat can be considered a significant route for the elimination of the com pound
from the environment.

In the sterile buffer solutions at pH 5.0, spirotet ramat was transformed to the major metabolites spirotetramat-photo-cyclopentyl, spirotetramat-phot o-hydroxymethyl, spirotetramat-photo-formyl and spirotetramat-photomethyl carbonate (all these meta bolites are considered photo-rearrangement products ). Maximum amounts formed were 42.9 %, 22.9 %, 11.5 % and 19.3 % of AR, respectively. Under dark conditions only the common hydrolysis product spiro tetramat-enol was observed. As demonstrated in a separate study (see following text), formation of t hese photo-rearrangement products, which has to be attributed to the highly artificial experimental co nditions, was not observed in sterilized natural wa ter at a pH
of 7.9.

### – 184 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

NH

O O O O O

H

Spirotetramat

NH

O O H

O H

Spirotetramat-photocyclopentyl Spirotetramat-photo2-methylcarbonate O O O

NH

O O H

NH

O O H

O H

Spirotetramat-photo2-hydroxy-methyl NH

O O H

O

Spirotetramat-photo2-formyl

CO 2



### Figure B.8.4.4.5-2: Proposed photolysis pathway of spirotetramat in sterile buffer solution (pH 5.0).

Under irradiation in sterilized natural water and p H 7.9, spirotetramat extensively degraded with a ha lf-life of 0.19 days (following SFO kinetics, R 2 

### = 1.00, net photolysis half-life corrected for hyd rolysis was 0.22 days). The experimental half-life (including photolysis an d hydrolysis) corresponds to 0.66, 0.50 and 0.62 da ys under environmental midday summer light in Athens ( Greece, EU), Phoenix (AZ, USA) and Edmonton (Alberta, Canada). In the dark control DegT 50 

### was 1.5 days (owing to hydrolysis, only) indicatin g that photodegradation of spirotetramat can be also consi dered a significant route for the elimination of th e compound from the environment under more alkaline c onditions.

Two major phototransformation products were formed besides the common hydrolysis product spirotetramat

enol: methoxy-cyclohexyl-aminocarboxylic acid (maxi mum 11.3 % of AR at study termination) and methoxycyclohexanone (17.5 % of AR), the latter a photolys is metabolite also observed in soil photolysis. No photorearrangement products were formed. Under dark cond itions only spirotetramat-enol was formed owing to hydrolysis.

The photolysis study in sterile natural water was c onducted to demonstrate that photo-rearrangement products observed under photolysis in sterile buffe r solution at pH 5 (Stupp, 2005a) are not likely to occur under more natural conditions. However, based on th e contrasting pH value of both photolysis studies ( pH 5.0 in sterile buffer solution vs. pH 7.9 in steril e natural water) no final conclusion on the occurre nce of photorearrangement product in sterilized natural water i n more acidic surface water can be drawn from the s tudy
results per se . However, based on the rapid degradation half-life (dark rate) of spirotetramat in water/sediment systems (pH 6.7 and 7.2) of approx. 0.8 days (Menke, 2006c, Röpke, 2006b) a significant

### – 185 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

formation of photo-rearrangement products as observ ed in Stupp (2005a), in which spirotetramat showed a dark hydrolysis rate of 26.2 days, is highly unlike ly to occur. Therefore, photo-rearrangement product s observed in sterile buffer solutions at pH 5 are no t considered relevant for further risk assessment.

NH

O O O O O

H

NH

O O H O

H

Spirotetramat Spirotetramat-enol

O H

O NH 2 O H O H

O

Methoxy-cyclohexanone Methoxy-cyclohexylamino carboxylic acid



### Figure B.8.4.4.5-3: Proposed photolysis pathway of spirotetramat in sterile natural water (pH 7.9).

In two separate studies, the quantum yields of the direct photochemical degradation of spirotetramat a nd spirotetramat-enol were investigated. These studies were carried out in sterile, unbuffered water (no pH measured) at a test concentration of 5 mg L-1 using a mercury immersion lamp TQ 150 (> 295 nm) and a uranyl oxalate actinometer. Mean degradation half l ives under these experimental conditions were 0.15 days (spirotetramat) and 1.39 days (spirotetramat-enol). No dark control samples were investigated to accou nt for hydrolysis processes. However, owing to the short e xposure time of the test solution (4 hrs) no signif icant degradation of spirotetramat owing to hydrolysis is expected. Spirotetramat-enol was shown to be hydrolytically stable from pH 4 – 9. Quantum yield for spirotetramat and spirotetramat-enol were calcu lated to be 0.00571 and 0.000252, respectively. Based on different arithmetic models (GC-SOLAR and Frank & Klöpffer) by means of resulting quantum yield and a dsorption, environmental direct phototransformation halflives of spirotetramat and spirotetramat-enol, in a range of approx. 0.5 – 1 days and 9 – 16 days, respectively, can be assessed for the period of mai n use (late spring to summer).

Biological degradation

Results of a readily biodegradability study indicate that spirotetramat is not readily biodegr adable.

Dark aerobic water/sediment studies were conducted with two contrasting (pH, texture) natural systems, ‘Hönniger Weiher’ and ‘Anglerweiher’, using [3- 14 

### C] and [5- 14 

### C] labelled spirotetramat. The ‘Hönniger Weiher’ test system represents a loam sediment with an orga nic carbon content of 4.4 %, a microbial activity o f 36 mg CO 2 

### h -1 

### kg -1 

### and a pH of 5.6 (CaCl 2 

### ). The ‘Anglerweiher’ test system is characterized by a loamy sand sediment with a pH of 6.8 (CaCl 2 

### ), with an organic carbon content of 0.99 % and a l ower microbial activity (10 mg CO 2 

### h -1 

### kg -1 

### ). In both systems, water and sediment stayed aerob ically throughout the test period.

Mineralization of spirotetramat to 14 

### CO 2 

### was low in the ‘Hönniger Weiher’ test system, acco unting for 11.0 % and 5.9 % of AR by study termination (120 DAT) usin g [3- 14 

### C] and [5- 14 

### C] labelled parent, respectively.
Formation of 14 

### CO 2 

### from [5- 14 

### C] labelled spirotetramat was approx. half indicati ng that this part of the molecule is less accessible to biodegradation than the [3- 14 

### C] position. Formation of 14 

### CO 2 

### was higher in the

### – 186 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

‘Anglerweiher’ test system accounting for 24.0 % an d 13.5 % of AR using [3- 14 

### C] and [5- 14 

### C] labelled parent, respectively.

Maximum formation of NER (mean value of both labels ) accounted for 36.4 % and 33.1 % of AR in the ‘Hönniger Weiher’ and Anglerweiher’, respectively. In the first test system, maximum of NER formation was observed by study termination (120 days), in the la tter system, maximum formation of NER was found by 91 DAT, slightly decreasing thereafter to 34.6 % of AR by study termination (120 days). Differences betwe en labels used were small in case of the ‘Hönninger We iher’ and insignificant in case of the ‘Angler Weih er’.

In the total system, a rapid decline of spirotetram at was observed with DegT 50 

### of 0.9 and 0.7 days in the ‘Hönninger Weiher’ and ‘Anglerweiher’ test system ( following SFO kinetics), respective DegT 90 

### values were 2.9 and 2.3 days. Dissipation in the water phase wa s calculated to be 0.8 and 0.7 in the ‘Hönninger We iher’ and ‘Anglerweiher’ test system. No distinct differe nces between labels used were observed.

Spirotetramat-enol (maximum in the entire system 99 .0 % of AR by 14 DAT, decreasing thereafter) and spirotetramat-ketohydroxy (maximum in the total sys tem 50.8 % of AR at study termination) were observe d as major (> 10 % of AR) metabolites. Based on multi -compartment modelling, spirotetramat-enol degraded with a mean DegT 50 

### of 47.3 days in the entire system (SFO kinetics, R 2 

### > 0.93). In comparison to soil systems, degradation of spirotetramat-enol is disti nct slower in aquatic systems. In contrast to the s oil (which is considered a triphasic system of soil, water and air), the water/sediment is just bi-phasic (suspen ded sediments and water containing more or less dissolv ed oxygen). Since the main path of spirotetramat-en ol degradation in soil is a surface catalyzed oxidatio n, it is reasonable that a significantly lower over all degradation in water/sediment system occurs (spirot etramat-enol is stable under conditions of sterile hydrolysis). This assumption is strongly supported by the anaerobic water/sediment study were spirotetramat-enol was observed to be stable at all

.

The major water/sediment metabolite spirotetramat-k etohydroxy was considered to be stable in both systems.

Both metabolites, spirotetramat-enol and spirotetra mat-ketohydroxy, were also shown to significantly dissipate into the sediment phase with a maximum oc currence of 36.6 % (by 60 DAT) and 27.8 % of AR (by study termination), respectively.

Other identified substances (spirotetramat-MA-amide , spirotetramat-oxo-enol isomer and spirotetramat-d ihydroxy) did not exceed 9.8 % of AR in the total sy stem individually, non identified radioactivity acc ounted for
maximum of 8.1 % of AR in sum.

### – 187 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

NH

O O O O O

H

NH

O O H O

H

Spirotetramat Spirotetramat-enol

NH

O O O H

OH

Spirotetramat-ketohydroxy

NH

O O H

OH O H

O

Spirotetramat-MA-amide NH

O O H O

H

OH

Spirotetramat-dihydroxy

NH

O O H

Spirotetramat-oxo-enol isomer O

CO 2 NER



### Figure B.8.4.4.5-4: Proposed degradation pathway of spirotetramat in aerobic water/sediment

systems (metabolites > 10 % of AR are indicated in bold).

One dark anaerobic water/sediment study is available, conducted with a test system (‘Lawre nce’) characterized by a clay loam sediment with a pH of 7.1 (CaCl 2 

### ) and an organic C content of 0.7 %.
Anaerobic conditions in the water and in the sedime nt were maintained throughout the study.

Under anaerobic water/sediment conditions spirotetr amat was completely transformed into its major metabolite spirotetramat-enol which can be consider ed stable under the experimental conditions. Spirotetramat-enol reached maximum amounts in the w ater of 80.5 % of AR by 14 DAT and in the sediment of 28.5 % of AR by 21 DAT hardly changing thereafte r. The degradation half-life of spirotetramat was calculated to be 2.8 days (R 2 

### = 0.99) in the entire system, respective DegT 90 

### was 9.3 days.

### – 188 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.4.4.5-1: Summary on DT 50 

### and DT 90 

### [days] for the dissipation and degradation of spirotetramat, spirotetramat-enol and spirotetramat -ketohydroxy in aerobic water/sediment studies. Water Sediment Total system Degradation Dissipation Degradation Degradation Compound Test system DegT 50 DegT 90 DT 50 DT 90 DegT 50 DegT 90 DegT 50 DegT 90

‘Hönninger Weiher’ nc nc 0.82 2.73 nc nc 0.86 2.85 Angler Weiher’ nc nc 0.74 2.45 nc nc 0.70 2.34 Spirotetramat

Geometric mean nc nc 0.78 2.59 nc nc 0.78 2.58

‘Hönninger Weiher’ nc nc nc nc nc nc 59.0 196 Angler Weiher’ nc nc nc nc nc nc 37.9 126Spirotetramatenol Geometric mean nc nc nc nc nc nc 47.3 157

‘Hönninger Weiher’ nc nc nc nc nc nc Stable Stable Angler Weiher’ nc nc nc nc nc nc Stable StableSpirotetramatketohydroxy Geometric mean nc nc nc nc nc nc Stable Stable

nc denotes not calculated



### Table B.8.4.4.5-2: Summary on DT 50 

### and DT 90 

### [days] for the dissipation and degradation of spirotetramat, spirotetramat-enol in anaerobic water/sediment studies. Water Sediment Total system Degradation Dissipation Degradation Degradation Compound Test

system DegT 50 DegT 90 DT 50 DT 90 DegT 50 DegT 90 DegT 50 DegT 90 Spirotetramat ‘Lawrence’ nc nc 2.8 9.2 nc nc 2.8 9.3 Spirotetramat-enol ‘Lawrence’ nc nc nc nc nc nc Stable Stable

nc denotes not calculated



### Table B.8.4.4.5-3: Summary on maximum occurrence [% of AR] of spirotetramat and metabolites in aerobic water/sediment studies (mean of labels used, data stated in brackets give day of maximum occurrence).

Compound Water Sediment Total Spirotetramat - 3.2 (1) - Spirotetramat-enol 78.8 (7) 36.6 (60) 97.5 (7) Spirotetramat-ketohydroxy 12.7 (120) 27.8 (120) 40.5 (120) Spirotetramat-MA-amide 7.0 (60) 1.8 (60) 8.7 (60) Spirotetramat-oxo-enol isomer 3.8 (91) 1.2 (120) 4.2 (120) Spirotetramat-di-hydroxy 2.2 (91) 3.8 (91) 6.0 (91)



### B.8.4.5 Degradation in the saturated zone (OECD Ann ex IIA, 7.9)

The degradation behaviour of spirotetramat in the s aturated zone has not been investigated in specific studies since it is not expected to reach such zone s after its use according to good agricultural prac tices. If at all needed it can be estimated from laboratory soil degradation studies as well as from the laboratory water/sediment studies performed under dark aerobic and anaerobic conditions. In these studies degradation of spirotetramat and its residues was c omparatively fast.

### – 189 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.5 Impact on water treatment procedures (OECD An nex IIIA, 9.6.4)

In the readily biodegradability study, the test sho wed that spirotetramat had no inhibitory effect on microbial activity in an activated sludge media. Furthermore, the activated sludge respiration test gave a spiro tetramat 3-hrs-EC 50 

### of > 10 000 mg L -1 

### . Results of modelling indicate that maximum predic ted concentrations of spirotetramat in surface water will not exceed 5.9 μg L -1 

### , indicating a large margin of safety. The concentration in groundwater does not exceed the ma ximum acceptable concentration of 0.1 μg L -1 

### . It can therefore be concluded that spirotetramat is unlike ly to impact adversely onto water treatment procedu res.

### – 190 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.6 Predicted environmental concentration in grou nd water for the active substance and

metabolites (PEC GW 

### ) (OECD Annex AIII, 9.6)

The risk assessment for the EU can be found in the APPENDIX.
The risk assessment for the PMRA and the US-EPA is not included in this monograph.

The Proposed Registration Decision – Spirotetramat (PRD2008-07), containing the detailed Canadian review and risk assessment, has been published by t he PMRA and can be found at the PMRA’s web site www.pmra-arla.gc.ca

.

The registration decision made by the U.S. EPA's Of fice of Pesticide Program is supported by the risk
assessments found at our Federal Docket website loc ated at: http://www.regulations.gov/fdmspublic/component/mai n?main=AdvancedDocket
The Docket ID is "EPA-HQ-OPP-2007-0475".

B.8.7 Predicted environmental concentration in surf ace water and sediment for the active

substance and metabolites (PEC SW 

### , PEC SED 

### ) (OECD Annex AIII, 9.7, 9.8)

The risk assessment for the EU can be found in the APPENDIX.
The risk assessment for the PMRA and the US-EPA is not included in this monograph.

The Proposed Registration Decision – Spirotetramat (PRD2008-07), containing the detailed Canadian review and risk assessment, has been published by t he PMRA and can be found at the PMRA’s web site www.pmra-arla.gc.ca

.

The registration decision made by the U.S. EPA's Of fice of Pesticide Program is supported by the risk
assessments found at our Federal Docket website loc ated at: http://www.regulations.gov/fdmspublic/component/mai n?main=AdvancedDocket
The Docket ID is "EPA-HQ-OPP-2007-0475".

### – 191 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.8 Fate and behaviour in the air

B.8.8.1 Rate and route of degradation in air (OECD Annex IIA, 7.10, IIIA, 9.9)

Chemical lifetime in the troposphere were calculate d for the compounds

• spirotetramat

• spirotetramat-enol

Reference: Calculation of the chemical lifetime of BYI08330 in the troposphere Author(s), year: Hellpointner, E., 2004a Study/Report number: M-092840-01-1, MEF-04/400 Guideline(s): None GLP: Calculations - not applicable Validity: Study considered acceptable

Material and methods: Based on the calculation according to Atkinson the chemical lifetime of the parent compound spirotetra mat (smiles notation: N1C(=O)C(c2c(C)ccc(C)c2)=C(OC(=O) OCC)C31CCC(OC)CC3) in the air was assessed by use of the software Atmospheric Oxidation Program ( AOPWIN, 1.91).

Findings:

Type of reaction Parameter Value Hydrogen abstraction [10 -12 cm 3 (molecule sec) -1 ] 26.3 Reaction with N, S and -OH [10 -12 cm 3 (molecule sec) -1 ] 5.50 Addition to triple bonds [10 -12 cm 3 (molecule sec) -1 ] 0.00 Addition to olefinic bonds [10 -12 cm 3 (molecule sec) -1 ] 38.5 Addition to aromatic rings [10 -12 cm 3 (molecule sec) -1 ] 5.87 Addition to fused rings [10 -12 cm 3 (molecule sec) -1 ] 0.00 Overall OH rate constant [10 -12 cm 3 (molecule sec) -1 ] 76.2 Reactions with OH - radicals

Half-life (12-hrs day, 1.5 × 10 6 OH - cm -3 ) [hrs] 1.69 Overall ozone rate constant [10 -17 cm 3 (molecule sec) -1 ] 2.1 Reactions with ozone Half-life (12-hrs day, 7 × 10 11 mol cm -3 ) [hrs] 13.1



### Conclusion: The photochemical (reaction with OH radicals) half- life of the parent compound spirotetramat in the troposphere was estimated by the method of Atkinson to be 1.69 hours (12-hrs day; 1.5 × 10 -6 

### OH - 

### cm -3 

### ).

Reviewer comments:
None

### – 192 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: Calculation of the chemical lifetime of BYI08330-en ol in the troposphere Author(s), year: Hellpointner, E., 2004b Study/Report number: M-092841-01-1, MEF-04/401 Guideline(s): None GLP: Calculations - not applicable Validity: Study considered acceptable

Material and methods: Based on the calculation according to Atkinson the chemical lifetime of the metabolite spirotetramat-e nol (smiles notation: N1C(=O)C(c2c(C)ccc(C)c2)=C(O)C31C CC(OC)CC3) in the air was assessed by use of the software Atmospheric Oxidation Program (AOPWIN, 1.9 1).

Findings:

Type of reaction Parameter Value Hydrogen abstraction [10 -12 cm 3 (molecule sec) -1 ] 24.7 Reaction with N, S and -OH [10 -12 cm 3 (molecule sec) -1 ] 5.64 Addition to triple bonds [10 -12 cm 3 (molecule sec) -1 ] 0.00 Addition to olefinic bonds [10 -12 cm 3 (molecule sec) -1 ] 38.5 Addition to aromatic rings [10 -12 cm 3 (molecule sec) -1 ] 5.87 Addition to fused rings [10 -12 cm 3 (molecule sec) -1 ] 0.00 Overall OH rate constant [10 -12 cm 3 (molecule sec) -1 ] 74.7 Reactions with OH - radicals

Half-life (12-hrs day, 1.5 × 10 6 OH - cm -3 ) [hrs] 1.72 Overall ozone rate constant [10 -17 cm 3 (molecule sec) -1 ] 2.1 Reactions with ozone Half-life (12-hrs day, 7 × 10 11 mol cm -3 ) [hrs] 13.1



### Conclusion: The photochemical (reaction with OH radicals) half- life of the metabolite spirotetramat-enol in the troposphere was estimated by the method of Atkinson to be 1.72 hours (12-hrs day; 1.5 × 10 -6 

### OH - 

### cm -3 

### ).

Reviewer comments:
None

### – 193 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.9 Predicted environmental concentration in the air (PEC A 

### )

The risk assessment for the EU can be found in the APPENDIX.
The risk assessment for the PMRA and the US-EPA is not included in this monograph.

The Proposed Registration Decision – Spirotetramat (PRD2008-07), containing the detailed Canadian review and risk assessment, has been published by t he PMRA and can be found at the PMRA’s web site www.pmra-arla.gc.ca

.

The registration decision made by the U.S. EPA's Of fice of Pesticide Program is supported by the risk
assessments found at our Federal Docket website loc ated at: http://www.regulations.gov/fdmspublic/component/mai n?main=AdvancedDocket
The Docket ID is "EPA-HQ-OPP-2007-0475".

### – 194 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.10 Definition of the residues (OECD Annex IIA, 7.11)

Soil: Spirotetramat, spirotetramat-enol, spirotetra mat-ketohydroxy, 4-methoxy-cyclohexanone (the

latter a soil photolysis product) Surface water: Spirotetramat, spirotetramat-enol, s pirotetramat-ketohydroxy, 4-methoxy-cyclohexanone,

4-methoxy-cyclohexylaminocarboxylic acid (the two l atter photolysis products) Sediment: Spirotetramat, spirotetramat-enol, spirot etramat-ketohydroxy Groundwater: Spirotetramat, spirotetramat-enol, spi rotetramat-ketohydroxy, spirotetramat-MA-amide,

4-methoxy-cyclohexanone (the latter a soil photolys is product) Air: Spirotetramat, spirotetramat-enol

B.8.11 Monitoring data (OECD Annex IIA, 7.12)

Plant protection products containing spirotetramat are not yet authorized for use. Accordingly, monito ring data concerning fate, behaviour and concentration i n the environment are not available so far.

B.8.12 Other/special studies (OECD Annex IIA, 7.13, AIII 9.10)

Reference: Position Paper – Spirotetramat (BYI08330) – Fate an d Behaviour (BCS Response to AGES Letter of Data Request 01 and 02 ( 2007-03-06 and 2007- 04-04) including requests from related meeting on M ay 04, 2007 and AGES Letters of Data Request 03, 04 and 05 (dated 2007-0 5-23, 2007-06-08 and 2007-06-19) Author(s), year: Hellpointner, E., 2007 Study/Report number: -, MEF-07/299 Guideline(s): Position paper - not applicable GLP: Position paper - not applicable Validity: Position paper considered acceptable

During evaluation of the submitted studies some unc lear aspects mainly concerning the dimerization of spirotetramat-enol and other issues were identified resulting in several data requests. In the present position paper the notifier tried to clarify the mentioned a spects.

Dimerization of spirotetramat-enol As an overview, the three existing spirotetramat ae robic soil metabolism studies with regard to the di fference in the used application method and the analytical c ondition are compared (Table B.8.11-1).

### – 195 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.12-1: Compilation of aerobic soil metabol ism studies.

Study Application method Processing Observation Con sequences

Spirotetramat study (Babczinski, 2005) ‘Hot spot’ a application (due to insolubility in water) Acidic processing Spirotetramatenol is to some degree unstable in acidic environment Application method

only usable for parent kinetics and metabolite identification  separate soil metabolism study was conducted with spirotetramat-enol

Spirotetramat-enol study (Babczinski, 2006b) Broadcast b application (realistic conditions) Alkaline processing (sufficiently soluble in water) Valid

spirotetramatenol degradation possible -

Outdoor study (Babczinski, 2006a) Broadcast b application with formulation (realistic conditions) Acidic processing - -

a Dosing the soil with a low volume of higher concent rated organic test solution (acetonitrile)
b Dosing the soil with a higher volume of low concent rated aqueous test solution



### In the Table B.8.11-2 an overview over the differen t soil metabolism and dissipation studies with rega rd to dimer formation is provided.

Table B.8.12-2: Compilation on the formation of spi rotetramat-enol dimers.

Test item Study type Scenario Application mode Tier Processing Max. observed dimer

value (% of applied)

Spirotetramat Aerobic soil

metabolism Lab ‘Hot spot’ (acetonitrile) Worst case Acidic 12.7

Spirotetramat-enol Aerobic soil

metabolism Lab Broadcast (water) Approaching

real case Alkaline 5.0

Spirotetramat Aerobic soil

metabolism Outdoor

(semifield) Broadcast

(formulation OD 100) Similar to

real case Acidic 1.5

Spirotetramat Terrestrial field dissipation Field Broadcast (formulation OD 100) Real world Acidic 0.2



### By comparing the two standard laboratory studies it was shown that the dimer formation was higher in t he parent study with ‘hot spot’ application than in th e spirotetramat-enol study with broadcast applicati on (more realistic environmental conditions).

Furthermore, within the parent study itself, much l ower portions of dimers (< 5 % of AR) were found in the
US soil ‘Molino’ which received a much lower concen trated treatment solution than the 3 EU soils (dime r formation up to 12.7 % of AR). The results of the s pirotetramat-enol study clearly show that spirotetr amatenol dimer 1 and 2 are minor transformation product s in soils. This holds also for the outdoor study w ith the formulated product.

Further, the reviewer noted that in the raw data fr om the US terrestrial field dissipation studies som e semiquantitative dimer data were extracted. Since these data could not be fully validated due to the lack of appropriate dimer reference standards it was not re ported in the original TFD report. However, they gi ve a clear indication of the significance of these dimer s under field conditions since soil extracts were a nalyzed by LC-MS also for m/z 600 signals at the retention tim es of both dimers (ROI 6 and ROI 7). The maximum di mer formation in field was below a concentration of 0.2 % of applied.

### – 196 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Concluding, the dimers were thoroughly investigated but are not taken into account for further risk assessment owing to their artificial formation obse rved in lab studies as well as due to their very lo w levels
under outdoor/field conditions. It is evident that under real case scenarios (broadcast application) e ven in the lab studies dimers do not play a significant role a nd are therefore considered not relevant.

Chemical pathway of spirotetramat-enol dimer format ion/degradation Based on the spirotetramat-enol dimers molecular we ight of m/z 600 (601 = M+H in ESI positive LC.MS) a nd their fragment m/z 301 (302 = M+H in ESI positive L C-MS/MS) in the LC-MS experiment, a dimeric structu re composed of two spirotetramat-enols was derived. Th e dimers molecular weight of 600 is two mass units (hydrogen) lower than the sum of two times m/z 301 (the molecular weight of the enol). Therefore, an oxidative dimerization of spirotetramat-enol or a d imerization of spirotetramat-enol with spirotetrama tketohydroxy under the formation of an ether linkage (spirotetramat-enol dimer 1) is proposed as the respective chemical reaction. As reaction partners, one spirotetramat-enol molecule in the ketoform of the ketoenol tautomers and one spirotetramat-enol in th e enolform of the ketoenol tautomers are proposed (Figure B.8.11-1). Alternatively, a dimerization of one spirotetramat-enol molecule with a spirotetram atketohydroxy molecule (formed by previous enol oxida tion) might be assumed. Further it was agreed that it is likely that a dimer like the one shown in the figur e below may hydrolyze to spirotetramat-enol and spirotetramat-ketohydroxy.

Figure B.8.12-1: Proposed chemical pathway of spiro tetramat-enol dimer formation/degradation

‘Immobile’ fraction of spirotetramat-enol - compara bility of behaviour under lab and field conditions
The kinetic evaluation of the metabolites was based on the laboratory study, in which the test item wa s applied at realistic local concentrations. Therefor e only low amounts of the dimers were observed (comparable to those under outdoor/field conditions ) and in this study the dimers did not play a signi ficant role. Therefore, the dimers were not considered in the kinetic evaluation. In the kinetic evaluation t he immobile fraction of spirotetramat-enol is defined as part of the observed spirotetramat-enol. The act ual value of the immobile fraction was established by f itting the SFO-RB (single first order reversible bi nding) model to observed spirotetramat-enol concentrations (for detail see kinetic evaluation of the metaboli tes, Röpke, 2006a). In the SFO-RB model the immobile (al so called protected) fraction represents a fraction of

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spirotetramat-enol, which is less available and thu s leads to the observed bi-phasic degradation behav iour. However, this portion was not based on any observed dimer concentration, but is part of the observed spirotetramat-enol concentrations.

To point out that a comparability of the degradatio n behaviour of spirotetramat and its metabolites un der laboratory conditions and under real outdoor condit ions is given, the US field dissipation trial were fully modelled with the data set suggested for environmen tal risk assessment (see study by Röpke and Ramanarayanan, 2007). The prediction of the soil re sidues was based on the kinetic parameters derived from laboratory metabolism studies and considered t he specific environmental conditions of the field t rials. In general, the results showed that the comparison ver ified the kinetic parameters for spirotetramat-enol and spirotetramat-ketohydroxy derived from the laborato ry study.

Overall conclusion on the dimerization issue Concluding, the dimers do not contribute major meta bolites. The occurrence of dimers does not signific antly influence the environmental behaviour of spirotetra mat and its major metabolites under field condition s. The derived environmental parameters are representative for the behaviour of spirotetramat and its major
metabolites under field conditions.

Comment (reviewer):

• The statements concerning the dimerization of spir otetramat-enol are considered acceptable.

• With regard to the reviewer’s data requests number 02, 03, 04 and 05 most of the statements of the notifier were included directly in the respective s tudies.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Physical-chemical data on spirotetramat metabolites spirotetramat-enol and spirotetramat-ketohydroxy w ere investigated in the following studies:

Reference: Water Solubility of BYI08330-enol (AE 1302944) at p H 5, pH 7 and pH 8 (Flask Method) Author(s), year: Eyrich, U., Bogdoll, B., 2006 Study/Report number: M-275829-01-2, PA-06/035 Guideline(s): Guideline 92/69/EEC, appendix A.6; OE CD-Guideline 105
GLP: Yes Validity: Study considered acceptable

Summary: The water solubility of spirotetramat-enol at pH 5, pH 7 and pH 8 has been determined according to the ‘flask method’ described in guideline 92/69/EEC, appendix A.6 and OECD-guideline 105.

Table B.8.12-3: Water solubility of spirotetramat-e nol.

System pH range a Solubility at 20°C [g L -1 ] SD [g L -1 ] RSD [%] buffer pH 5 4.98 0.09 0.01 14.3 buffer pH 7 6.97 – 6.99 2.7 0.003 0.1 buffer pH 8 b 7.82 – 7.83 28 0.3 1.0

a pH-range of 3 experiments
b Due to the strong acidic character of the test item the buffer capacity of the used buffer pH 9 was not sufficient. Therefore, the pH value of the saturated solution declined to pH 8.



### Reviewer comments:
None

Reference: Partition Coefficients 1-Octanol/Water of BYI08330- enol (AE 1302944) at pH 5, pH 7 and pH 9 (Shake flask method) Author(s), year: Bogdoll, B., Eyrich, U., 2006 Study/Report number: M-276091-01-2, PA-06/036 Guideline(s): EEC Test Guideline 92/69, A.8.; OECD Test Guideline 107
GLP: Yes Validity: Study considered acceptable

Summary: The partition coefficients 1-octanol/water of the t est item spirotetramat-enol at pH 5, pH 7 and pH 9 at room temperature have been determined according to the ‘ flask-shaking’ method described in EEC Test Guideli ne 92/69, A.8. and OECD Test Guideline 107.

Table B.8.12-4: Partition coefficients (1-octanol/w ater) of spirotetramat-enol.

Parameter pH 5 pH 7 pH 9 Log(P OW ) 2.0 0.3 -1.3
P OW 109 2.1 0.05



### Reviewer comments:
None

### – 199 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Reference: BYI08330-enol (AE 1302944): Determination of the Di ssociation Constant (Spectrophotometric Screening Method) Author(s), year: Bogdoll, B., Lemke, G., Kaußmann., M. 2006 Study/Report number: M-274084-01-2, AF06/031 Guideline(s): Based on OECD Test Guideline 112 GLP: None GLP screening test Validity: Study considered acceptable

Summary: The dissociation constant of spirotetramat-enol was determined using a spectrophotometric method, base d on the OECD-Guideline 112. The experimental work ha s been performed by Dr. M. Kaußmann (PT - Analytics Monheim). An instrument for pK a 

### screening measurements from Sirius Analytical Inst ruments Ltd (UK) was used (type GLpK a 

### with D-Pas, Dip Probe Absorption Spectroscopy).

The dissociation constant (pK a 

### ) of spirotetramat-enol was 5.2.

Reviewer comments:
None

Reference: Water Solubility of BYI 08330-cis-ketohydroxy in Di stilled Water (Flask Method) Author(s), year: Bogdoll, B., Lemke, G., 2005a Study/Report number: M-268425-01-2, PA05/099 Guideline(s): Guideline 92/69/EEC, appendix A.6; OECD-Guideline 1 05
GLP: Yes Validity: Study considered acceptable

Summary: The water solubility of spirotetramat-ketohydroxy h as been determined in distilled water according to the ‘flask method’ described in guideline 92/69/EEC, ap pendix A.6 and OECD-guideline 105.

Table B.8.12-5: Water solubility of spirotetramat-k etohydroxy in distilled water. Solubility at 20°C [g L -1 ] SD [g L -1 ] RSD [%]

0.228 0.0042 1.8



### Reviewer comments:
None

Reference: Partition Coefficient 1-Octanol/Water of BYI08330-c is-ketohydroxy (HPLC Method) Author(s), year: Bogdoll, B., Lemke, G., 2005b Study/Report number: M-268428-01-2, PA05/098 Guideline(s): 92/69/EEC A.8, OECD TG 117, OPPTS 830.7570
GLP: Yes Validity: Study considered acceptable

Summary:

### – 200 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

The partition coefficient (1-octanol/water) of spir otetramat-ketohydroxy was determined according to O ECD test guideline 117, EEC-guideline 92/69/EEC A.8 and EPA guideline OPPTS 830.7570, HPLC-method. Nine calibration substances were injected into a HPLC-sy stem under the same analytical conditions (column temperature 40 °C) as the test item. Calibration cu rves were created by using the measured retention t imes (log(k’) values) and the known log(P OW 

### ) values of the calibration substances for linear r egression.

The measured log(k’) values of the test item were w ithin the calibrated log(k’) range. From the result ed calibration curves and their equations the log(P OW 

### ) of the substance has been interpolated.

Table B.8.12-6: Partition coefficients (1-octanol/w ater) of spirotetramat-ketohydroxy.

Parameter pH 7 Log(P OW ) 1.3
P OW 20



### Reviewer comments:
None

Reference: BYI08330-cis-ketohydroxy (AE 1422479): Determinatio n of the Dissociation Constant (Spectrophotometric Screening Method) Author(s), year: Bogdoll, B., Lemke, G., Kaußmann., M., 2005 Study/Report number: M-263603-01-2, AF05/098 Guideline(s): Based on OECD Test Guideline 112 GLP: None GLP screening test Validity: Study considered acceptable

Summary: The dissociation constant of spirotetramat-ketohydr oxy was determined by using a spectrophotometric
method based on the OECD Test Guideline 112. The ex perimental work has been performed by Dr. M. Kaußmann (PT - Analytics Monheim). An instrument fo r pK a 

### screening measurements from Sirius Analytical Instruments Ltd (UK) was used (type GLpK a 

### with D-Pas, Dip Probe Absorption Spectroscopy).

The dissociation constant (pK a 

### ) of spirotetramat-ketohydroxy was 11.0.

Reviewer comments:
None

### – 201 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.13 References relied on

Annex point/ reference number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant) Published or not Data Protection Claimed

Y/N-R/NR Owner

OECD Annex II Data and Information

KIIA, 7.1.1 KIIA, 7.2.1 Babczinski, P. 2005 Aerobic degradation/metabolism of BYI8330 in four different soils Bayer CropScience AG, Report No.: MEF-04/169, Edition Number: M-256849-02-2 Date: 07.07.2005, Amended: 05.05.2006 GLP, unpublished Yes BCS

KIIA, 7.1.1 Babczinski, P. 2006a Outdoor metabolism of [azaspirodecenyl-3-
14 C]BYI08330 in two soils Bayer CropScience AG, Report No.: MEF-06/041, Edition Number: M-270597-01-2 Date: 03.04.2006 GLP, unpublished Yes BCS

KIIA, 7.1.1, KIIA, 7.2.3 Babczinski, P. 2006b [Azaspirodecenyl-3- 14 C]- and [azaspirodecenyl-5- 14 C]-labelled BYI08330-cisenol: Comparative aerobic soil metabolism/degradation in four soils Bayer CropScience AG, Report No.: MEF-05/157, Edition Number: M-269304-01-2 Date: 17.01.2006 GLP, unpublished Yes BCS

KIIA, 7.1.2, KIIA, 7.2.4 Menke, U. 2006a BYI08330: Anaerobic soil metabolism Bayer CropScience AG, Report No.: MEF-05/515, Edition Number: M-270739-01-2 Date: 03.05.2006 GLP, unpublished Yes BCS

KIIA, 7.1.3 Heinemann,

O. 2005a [Azaspirodecenyl-3- 14 C]- and [azaspirodecenyl-5- 14 C]BYI08330: Phototransformation on soil Bayer CropScience AG, Report No.: MEF-04/481, Edition Number: M-252907-01-2 Date: 25.04.2005 GLP, unpublished Yes BCS

KIIA, 7.2.1, KIIA, 7.1.1 Babczinski, P. 2005 Aerobic degradation/metabolism of BYI8330 in four different soils Bayer CropScience AG, Report No.: MEF-04/169, Edition Number: M-256849-02-2 Date: 07.07.2005, Amended: 05.05.2006 GLP, unpublished Yes BCS

KIIA, 7.2.1 Ramanarayan

an, T.; Roepke, B. 2006 Degradation of BYI08330 in four soils under aerobic conditions: kinetic evaluation Bayer CropScience AG, Report No.: MEF-05/249, Edition Number: M-277096-01-2 Date: 30.08.2006 Non GLP, unpublished Yes BCS

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Annex point/ reference number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant) Published or not Data Protection Claimed

Y/N-R/NR Owner

KIIA, 7.2.3 KIIA, 7.1.1 Babczinski, P. 2006b [Azaspirodecenyl-3- 14 C]- and [azaspirodecenyl-5- 14 C]-labelled BYI08330-cisenol: Comparative aerobic soil metabolism/degradation in four soils Bayer CropScience AG, Report No.: MEF-05/157, Edition Number: M-269304-01-2 Date: 17.01.2006 GLP, unpublished Yes BCS

KIIA, 7.2.3 Roepke, B. 2006a Kinetic evaluation of labo ratory soil degradation studies of BYI08330-Enol, BYI08330-ketohydroxy, BYI08330-MA-amide Bayer CropScience AG, Report No.: MEF-06/199, Edition Number: M-277178-03-1 Date: 30.08.2006, Amended: 23.08.2007 Non GLP, unpublished Yes BCS

KIIA, 7.2.3 Dehner, D.;

Heinemann, O. 2006 [Carbonyl- 14 C] 4-methoxycyclohexanone: Aerobic soil degradation in three soils Bayer CropScience AG, Report No.: MEF-05/485, Edition Number: M-269022-02-2 Date: 28.02.2006, Amended: 18.07.2006 GLP, unpublished Yes BCS

KIIA, 7.2.4, KIIA, 7.1.2 Menke, U. 2006a BYI08330: Anaerobic soil metabolism Bayer CropScience AG, Report No.: MEF-05/515, Edition Number: M-270739-01-2 Date: 03.05.2006 GLP, unpublished Yes BCS

KIIA, 7.3.1 Netzband, D.

J. 2006a In house laboratory validation of an analytic al method for the determination of BYI08330 and its metabolites BYI08330-enol, BYI08330- ketohydroxy and BYI08330-MA-amide in soil and sediment by LC/MS/MS Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: RAFNX012, Edition Number: M-277365-01-1 Method Report No US: RAFNX012 Date: 29.08.2006 GLP, unpublished Yes BCS

KIIA, 7.3.1 Lenz, Mark 2006a Terrestrial field dissip ation of BYI08330 in New York soil, 2004 Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: MEFNY004, Edition Number: M-277191-01-1 Date: 05.09.2006 GLP, unpublished Yes BCS

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Annex point/ reference number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant) Published or not Data Protection Claimed

Y/N-R/NR Owner

KIIA, 7.3.1 Lenz, Mark 2006b Terrestrial field dissip ation of BYI08330 in Florida soil, 2004 Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: MEFNX055, Edition Number: M-277184-01-1 Date: 05.09.2006 GLP, unpublished Yes BCS

KIIA, 7.3.1 Lenz, Mark 2006c Terrestrial field dissip ation of BYI08330 in California soil, 2004 Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: MEFNY002, Edition Number: M-277186-01-1 Date: 05.09.2006 GLP, unpublished Yes BCS

KIIA, 7.3.1 Lenz, Mark 2006d Terrestrial field dissip ation of BYI08330 in Washington soil, 2004 Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: MEFNY003, Edition Number: M-277189-01-1 Date: 05.09.2006 GLP, unpublished Yes BCS

KIIA, 7.3.1 Netzband, D.

J. 2006b Stability of AE 0317309 and AE B197555 in soil during frozen storage, USA, 2005 (reported through a maximum of 320 days storage) Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: RAFNX018, Edition Number: M-277371-01-1 Date: 29.08.2006 GLP, unpublished Yes BCS

KIIA, 7.3.1 Brumhard, B. 2006 Independent laboratory va lidation of method FN-002-S05-02 for the determination of BYI08330 and its metabolites BYI08330-enol,BYI08330-keto-hydroxy and BYI08330-MAamide in soil and sediment by LS/MS/MS Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: MR-06/037, Edition Number: M-276038-01-2 Date: 14.08.2006 GLP, unpublished Yes BCS

KIIA, 7.4.1 Haas, M. 2005 BYI08330: Adsorption/desorpti on in five soils Bayer CropScience AG, Report No.: MEF-04/373, Edition Number: M-266755-01-2 Date: 21.10.2005 GLP, unpublished Yes BCS

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Annex point/ reference number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant) Published or not Data Protection Claimed

Y/N-R/NR Owner

KIIA, 7.4.2 Hein, W. 2005a Adsorption/desorption of BYI 08330-cis-enol in five different soils Rheinland-Pfalz AgroScience GmbH, Neustadt, Germany Bayer CropScience AG, Report No.: IM2000, Edition Number: M-267858-01-2 Date: 05.09.2005 GLP, unpublished Yes BCS

KIIA, 7.4.2 Hein, W. 2005b 14 C-BYI08330-ketohydroxy: Adsorption/desorption in five soils Rheinland-Pfalz AgroScience GmbH, Neustadt, Germany Bayer CropScience AG, Report No.: IM2001, Edition Number: M-267955-02-2 Date: 09.09.2005, Amended: 30.08.2006 GLP, unpublished Yes BCS

KIIA, 7.4.2 Simmonds,

M.; Early, E. 2005 [ 14 C]-BYI08330-MA-amide: Adsorption to and desorption from five soils Battelle UK Ltd.; Ongar; United Kingdom Bayer CropScience AG, Report No.: CX/04/070, Edition Number: M-263686-01-2 Date: 07.09.2005 GLP, unpublished Yes BCS

KIIA, 7.4.4 Babczinski, P. 2006c BYI08330-enol: Soil colu mn leaching Bayer CropScience AG, Report No.: MEF-05/356, Edition Number: M-270280-02-2 Date: 24.03.2006, Amended: 22.05.2006 GLP, unpublished Yes BCS

KIIA, 7.5, KIIA, 2.9.1 Heinemann,

O. 2004a [Azaspirodecenyl-3- 14 C]- and [azaspirodecenyl-5- 14 C]BYI08330: Hydrolytic degradation Bayer CropScience AG, Report No.: MEF-04/176, Edition Number: M-093124-01-2 Date: 08.09.2004 GLP, unpublished Yes BCS

KIIA, 7.5 Heinemann,

O. 2004b [Azaspirodecenyl-3- 14 C]- and [azaspirodecenyl-5- 14 C]BYI08330-enol: Hydrolytic degradation Bayer CropScience AG, Report No.: MEF-04/311, Edition Number: M-242999-01-2 Date: 03.12.2004 GLP, unpublished Yes BCS

KIIA, 7.6 Stupp, H. - P. 2005a BYI08330: Phototransforma tion of BYI08330 in sterile water Bayer CropScience AG, Report No.: MEF-05/206, Edition Number: M-266695-01-2 Date: 15.11.2005 GLP, unpublished Yes BCS

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Annex point/ reference number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant) Published or not Data Protection Claimed

Y/N-R/NR Owner

KIIA, 7.6, KIIA, 2.9.4 Stupp, H. - P. 2005b [Azaspirodecenyl-3- 14 C]BYI08330 and [Azaspirodecenyl-5- 14 C]BYI08330: Phototransformation in natural water Bayer CropScience AG, Report No.: MEF-05/262, Edition Number: M-266753-01-2 Date: 15.11.2005 GLP, unpublished Yes BCS

KIIA, 7.6, KIIA, 2.9.3, KIIA, 2.9.4 Heinemann,

O. 2004c BYI08330: Determination of the quantum yield and assessment of the environmental half-life of the direct photodegradation in water Bayer CropScience AG, Report No.: MEF-04/080, Edition Number: M-092941-01-2 Date: 28.09.2004 GLP, unpublished Yes BCS

KIIA, 7.6 Heinemann,

O. 2005b BYI08330-enol: Determination of the quantum yield and assessment of the environmental half-life of the direct photodegradation in water Bayer CropScience AG, Report No.: MEF-04/438, Edition Number: M-243787-01-2 Date: 10.01.2005 GLP, unpublished Yes BCS

KIIA, 7.7 Weyers, A. 2005 BYI 08330 - Biodegradation Bayer Industry Services, Leverkusen, Germany Bayer CropScience AG, Report No.: 2005/0077/01, Edition Number: M-263287-01-1 Date: 16.11.2005 GLP, unpublished Yes BIS

KIIA, 7.8.3 Menke, U. 2006c BYI08330: Aerobic aquatic metabolism Bayer CropScience AG, Report No.: MEF-04/511, Edition Number: M-269307-01-2 Date: 29.03.2006 GLP, unpublished Yes BCS

KIIA, 7.8.3 Roepke, B. 2006b Kinetic evaluation of the aerobic aquatic metabolism of BYI08330, BYI08330-enol and BYI08330-ketohydroxy in water sediment systems Bayer CropScience AG, Report No.: MEF-06/279, Edition Number: M-277415-01-1 Date: 30.08.2006 Non GLP, unpublished Yes BCS

KIIA, 7.8.3 Shepherd, J.;

Arthur, E. L.; Dominic, A. 2005 BYI08330[azaspirodecenyl-3- 14 C]: Anaerobic aquatic metabolism Bayer CropScience, Stilwell, KS, USA Bayer CropScience AG, Report No.: MEFNX015, Edition Number: M-261943-01-1 Date: 06.12.2005 GLP, unpublished Yes BCS

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Annex point/ reference number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant) Published or not Data Protection Claimed

Y/N-R/NR Owner

KIIA, 7.10, KIIA, 2.10 Hellpointner,

E. 2004a Calculation of the chemical lifetime of BYI 08330 in the troposphere Bayer CropScience AG, Report No.: MEF-04/400, Edition Number: M-092840-01-1 Date: 14.09.2004 Non GLP, unpublished Yes BCS

KIIA, 7.10 Hellpointner,

E. 2004b Calculation of the chemical lifetime of BYI 08330-enol in the troposphere Bayer CropScience AG, Report No.: MEF-04/401, Edition Number: M-092841-01-1 Date: 21.09.2004 Non GLP, unpublished Yes BCS

KIIA, 7.13 Hellpointner,

E. 2007 Position Paper: Spirotetramat (BYI08330) – Fate and Behaviour Bayer CropScience Deutschland GmbH, Frankfurt am Main, Germany Bayer CropScience AG, Report No.: MEF-07/299, Edition Number: - Date: 06.07.2007 GLP, unpublished Yes BCS

KIIA, 7.13 Eyrich, U.;

Bogdoll, B. 2006 Water solubility of BYI 08330-enol (AE 1302944) at pH 5, pH 7 and pH 8 (Flask Method) Bayer CropScience GmbH, Frankfurt am Main, Germany Bayer CropScience AG, Report No.: PA06/035, Edition Number: M-275829-01-2 Date: 11.08.2006 GLP, unpublished Yes BCS

KIIA, 7.13 Bogdoll, B.;

Eyrich, U. 2006 Partition coefficients 1-Octanol / water of BYI 08330-enol (AE 1302944) at pH 5, pH 7 and pH 9 (Shake flask method) Bayer CropScience Deutschland GmbH, Frankfurt am Main, Germany Bayer CropScience AG, Report No.: PA06/036, Edition Number: M-276091-01-2 Date: 17.08.2006 GLP, unpublished Yes BCS

KIIA, 7.13 Bogdoll, B.;

Lemke, G.; Kaussmann, M. 2006 BYI 08330-enol (AE 1302944) - Determination of the dissociation constant (spectrophotometric screening method) Bayer CropScience GmbH, Frankfurt am Main, Germany Bayer CropScience AG, Report No.: AF06/031, Edition Number: M-274084-01-2 Date: 05.07.2006 Non GLP, unpublished Yes BCS

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Annex point/ reference number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant) Published or not Data Protection Claimed

Y/N-R/NR Owner

KIIA, 7.13 Bogdoll, B.;

Lemke, G. 2005a Water solubility of BYI 08330-cis-ketohydroxy in distilled water (flask method) Siemens AG, Frankfurt am Main, Germany Bayer CropScience AG, Report No.: PA05/099, Edition Number: M-268425-01-2 Date: 06.09.2005 GLP, unpublished Yes BCS

KIIA, 7.13 Bogdoll, B.;

Lemke, G. 2005b BYI 08330-cis-ketohydroxy partition coefficien t 1-Octanol/water (HPLC-method) Bayer CropScience GmbH, Frankfurt am Main, Germany Bayer CropScience AG, Report No.: PA05/098, Edition Number: M-268428-01-2 Date: 06.09.2005 GLP, unpublished Yes BCS

KIIA, 7.13 Bogdoll, B.;

Lemke, G.; Kaussmann, M. 2005 BYI 08330-cis-ketohydroxy (AE 1422479) Determination of the dissociation constant (spectrophotometric screening method) - Code: PBF-2005-0070-TOX-06867 Bayer CropScience GmbH, Frankfurt am Main, Germany Bayer CropScience AG, Report No.: AF05/098, Edition Number: M-263603-01-2 Date: 16.12.2005 Non GLP, unpublished Yes BCS

OECD Annex III Data and Information KIIIA, 9.4, KIIIA, 9.5 Roepke, B. 2006a Predicted environmental concentratio ns of spirotetramat and its main metabolites in soil (PECsoil); Use in citrus in Europe. Bayer CropScience AG, Report No.: MEF-06/282, Edition Number: M-277210-01-1 Date: 20.08.2006, Non GLP, unpublished Yes BCS

KIIIA, 9.4, KIIIA, 9.5 Roepke, B. 2006b Predicted environmental concentratio ns of spirotetramat and its main metabolites in soil (PECsoil); Use in leafy vegetables in Europe. Bayer CropScience AG, Report No.: MEF-06/387, Edition Number: M-277222-01-1 Date: 20.08.2006, Non GLP, unpublished Yes BCS

KIIIA, 9.4, KIIIA, 9.5 Roepke, B. 2007a Predicted environmental concentratio ns of 4- methoxy cyclohexanone in soil (PECsoil). Bayer CropScience AG, Report No.: MEF-07/478, Edition Number: M-295000-01-1, Date: 15.11.2007 Non GLP, unpublished Yes BCS

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Annex point/ reference number Author(s) Year Title Source (where different from company) Company, Report No GLP or GEP status (where relevant) Published or not Data Protection Claimed

Y/N-R/NR Owner

KIIIA, 9.6, KIIIA, 9.6.1, KIIIA, 9.6.2 Roepke, B. 2006c Predicted environmental concentratio ns of spirotetramat and its main soil metabolites in groundwater recharge (PECgw) based on calculations with FOCUS PEARL; Use in citrus in Europe. Bayer CropScience AG, Report No.: MEF-06/280, Edition Number: M-277121-02-2, Date: 30.08.2006, Amended: 12.06.2007, Non GLP, unpublished Yes BCS

KIIIA, 9.6, KIIIA, 9.6.1, KIIIA, 9.6.2 Roepke, B. 2006d Predicted environmental concentratio ns of spirotetramat and its main soil metabolites in groundwater recharge (PECgw) based on calculations with FOCUS PEARL; Use in leafy vegetables in Europe. Bayer CropScience AG, Report No.: MEF-06/385, Edition Number: M277164-02-2, Date: 30.08.2006, Amended: 12.06.2007, Non GLP, unpublished Yes BCS

KIIIA, 9.6, KIIIA, 9.6.1, KIIIA, 9.6.2 Roepke, B. 2007b Predicted environmental concentratio ns of 4- methoxy cyclohexanone in groundwater recharge (PECgw) based on calculations with FOCUS PEARL. Bayer CropScience AG, Report No.: MEF-07/477, Edition Number: M-294996-01-1, Date: 15.11.2007, Non GLP, unpublished Yes BCS

KIIIA, 9.7, KIIIA, 9.8 Roepke, B. 2006e Predicted environmental concentratio ns of spirotetramat and its main metabolites in surface water and sediment (PECsw/sed) calculated according to FOCUS; Use in citrus in Europe. Bayer CropScience AG, Report No.: MEF-06/281, Edition Number: M-277190-01-1 Date: 30.08.2006, Non GLP, unpublished Yes BCS

KIIIA, 9.7, KIIIA, 9.8 Roepke, B. 2006f Predicted environmental concentratio ns of spirotetramat and its main metabolites in surface water and sediment (PECsw/sed) calculated according to FOCUS; Use in leafy vegetables in Europe. Bayer CropScience AG, Report No.: MEF-06/386, Edition Number: M-277176-01-1 Date: 30.08.2006, Non GLP, unpublished Yes BCS

### – 209 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008



# APPENDIX



# Spirotetramat

B.8 Fate and behaviour in the environment

Risk assessment for the EU (Annex III)

### – 210 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.3 Predicted environmental concentration in soil (PEC S 

### ) for the active substance and

relevant metabolites (OECD Annex IIIA, 9.4, 9.5)

Reference: Predicted environmental concentrations of spirotetr amat and its main metabolites in soil (PEC soil 

### ): Use in citrus in Europe Author(s), year: Röpke, B., 2006a Study/report number: M-277210-01-1/MEF-06/282 Guideline(s): Calculations – Not applicable GLP: Calculations – Not applicable Deviations: Calculations – Not applicable Validity: Study considered acceptable

and

Reference: Predicted environmental concentrations of spirotetr amat and its main metabolites in soil (PEC soil 

### ): Use in leafy vegetables in Europe Author(s), year: Röpke, B., 2006b Study/report number: M-277222-01-1/MEF-06/387 Guideline(s): Calculations – Not applicable GLP: Calculations – Not applicable Deviations: Calculations – Not applicable Validity: Study considered acceptable

and

Reference: Predicted Environmental Concentrations of 4-methoxy cyclohexanone in Soil (PEC soil 

### ) Author(s), year: Röpke, B., 2007a Study/report number: M-295000-01-1/MEF-07/478 Guideline(s): Calculations – Not applicable GLP: Calculations – Not applicable Deviations: Calculations – Not applicable Validity: Study considered acceptable

Material and methods:
The initial PEC S 

### [mg kg -1 

### ] of the active ingredient from a single applicatio n depends on the application rate A [g ha -1 

### ] of the compound, on the crop interception C [%], on the bulk density BD of the dry soil [g cm -3 

### ] (standard value 1.5 g cm -3 

### ) and on the assumed mixing depth d [cm] (standard value 5 cm), according to

PEC S,init 

### = A × (1 – 0.01 × C) / (100 × BD × d)

In single application scenarios, the initial PECS i s equal to the overall maximum. Based on the maximu m PEC S 

### and assuming simple first-order kinetics (SFO) wit h degradation rate k [d -1 

### ] (k = ln(2)/DT 50 

### ), the soil concentration over time [d] are given by

PEC S 

### (t) = PEC S,max 

### × e -k × t

### – 211 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

For a comparison with effect endpoints from long-te rm (chronic) ecotoxicological studies, it is someti mes more appropriate to use time-weighted average (TWA) exposure concentrations. For SFO kinetics, the TWA concentrations are given by

TWA S 

### (t) = PEC S,max 

### × 1 / (k × t) × (1 – e -k × t 

### )

For the calculation of the maximum PEC S 

### of a metabolite, the maximum amount of a metabolit e, X Met,s,max 

### [% of applied parent compound] that was observed in a degradation study, and the molar masses of parent, M P 

### , and M Met 

### [g mol -1 

### ], must be taken into account:

PEC Met,s,init 

### = A × (1 – 0.01 × C) × 1 / (100 × BD × d) × 0.01 × X Met,s,max 

### × (M Met 

### / M P 

### )

PEC S 

### and TWA S 

### values of a metabolite are calculated correspondin gly to the parent, using the degradation rate of the metabolite, k Met



### .

Table B.8.3-1: Application data of spirotetramat ap plied as OD 150 formulation.

Crop Citrus Leafy vegetables Application rate [g ha -1 ] 288 a 72 Number of applications 2 2 Interval [days] 21 14 Crop interception [%] 70 b 70 c

BBCH code 71 – 78 42 - 43 Soil depth [cm] 5 5 Soil density [g cm -3 ] 1.5 1.5

a Maximum application rate assuming 96 g ai ha -1 m -1 tree height (maximum tree height 3 m)
b FOCUS default value for all stages of citrus (SANCO/32 1/2000 rev. 2)

c FOCUS default value for this crop and BBCH code (SANCO/3 21/2000 rev. 2)

### – 212 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.3-2: Input parameter for spirotetramat an d metabolites. Compound Parameter Value Remarks

DegT 50 [days] 0.23 Worst case, lab, SFO, normalized, n = 4 Spirotetramat Molecular weight [g mol -1 ] 373.45

DegT 50 [days] 2.64 Worst case, lab (spirotetramat-enol degradation study), SFO recalculated from FOMC-DT 90 divided by 3.32, normalized, n = 4 Maximum occurrence [%] 100.0 Unknown, worst case as sumptionSpirotetramatenol

Molecular weight [g mol -1 ] 301.38

DegT 50 [days] 12.1 Worst case, lab (spirotetramat-enol degradation study), SFO, normalized, n = 4 Maximum occurrence [%] 24.0 Spirotetramat-enol degra dation study, labSpirotetramatketohydroxy Molecular weight [g mol -1 ] 317.38

DegT 50 [days] 0.6 Worst case, lab, SFO, non-normalized, n = 3 Maximum occurrence [%] 10.0 Soil photolysis, lab4-Methoxycyclohexanone Molecular weight [g mol -1 ] 128.17



### Findings:

Table B.8.3-3: Calculated PEC S 

### values [mg kg -1 

### ] and TWA S 

### values [mg kg -1 

### ] for spirotetramat and major metabolites in soil (multiple application ).

Spirotetramat Spirotetramatenol Spirotetramatketohydroxy 4-Methoxycyclohexanone Crop Days after

last application a PEC S

[mg kg -1 ] TWA S [mg kg -1 ] PEC S

[mg kg -1 ] TWA S [mg kg -1 ] PEC S

[mg kg -1 ] TWA S [mg kg -1 ] PEC S

[mg kg -1 ] TWA S [mg kg -1 ]
0 0.115 - 0.093 - 0.031 - 0.004 - 1 0.006 0.036 0.072 0.082 0.029 0.030 0.001 0.003
2 < 0.001 0.019 0.055 0.073 0.027 0.029 < 0.001 0.002

4 < 0.001 0.010 0.033 0.058 0.024 0.027 < 0.001 0.001

7 < 0.001 0.005 0.015 0.043 0.020 0.025 < 0.001 0.001

14 < 0.001 0.003 0.002 0.025 0.014 0.021 < 0.001 < 0.0 01
21 < 0.001 0.002 < 0.001 0.017 0.009 0.018 < 0.001 < 0 .001
28 < 0.001 0.001 < 0.001 0.013 0.006 0.015 < 0.001 < 0 .001
42 < 0.001 0.001 < 0.001 0.008 0.003 0.012 < 0.001 < 0 .001
50 < 0.001 0.001 < 0.001 0.007 0.002 0.010 < 0.001 < 0 .001 Citrus

100 < 0.001 0.001 < 0.001 0.004 < 0.001 0.005 < 0.001 < 0.001

0 0.029 - 0.024 - 0.009 - 0.001 - 1 0.001 0.009 0.018 0.021 0.008 0.008 < 0.001 0.001
2 < 0.001 0.005 0.014 0.018 0.008 0.008 < 0.001 < 0.0 01

4 < 0.001 0.002 0.008 0.015 0.007 0.008 < 0.001 < 0.0 01

7 < 0.001 0.001 0.004 0.011 0.006 0.007 < 0.001 < 0.0 01

14 < 0.001 0.001 < 0.001 0.006 0.004 0.006 < 0.001 < 0 .001
21 < 0.001 < 0.001 < 0.001 0.004 0.003 0.005 < 0.001 < 0.001
28 < 0.001 < 0.001 < 0.001 0.003 0.002 0.004 < 0.001 < 0.001
42 < 0.001 < 0.001 < 0.001 0.002 0.001 0.003 < 0.001 < 0.001
50 < 0.001 < 0.001 < 0.001 0.002 < 0.001 0.003 < 0.001 < 0.001 Leafy vegetables

100 < 0.001 < 0.001 < 0.001 0.001 < 0.001 0.001 < 0.001 < 0.001

a Last application always represents the worst case for all compounds



### Conclusion:
Calculated maximum PEC S 

### values for spirotetramat, spirotetramat-enol, spir otetramat-ketohydroxy and 4- methoxy-cyclohexanone (the latter a soil photolysis metabolite) were 0.115, 0.093, 0.031 and 0.004 mg kg -1



### , respectively, for use of 150 OD in citrus and 0.029 , 0.024, 0.009 and 0.001 mg kg -1 

### , respectively, for use of 150 OD in leafy vegetables.

### – 213 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Comments (RMS):

• The notifier also presented PEC S 

### calculations for spirotetramat-MA-amide. Since thi s metabolite
does not exceed 10 % of AR in soil and does not sho w a steady increase towards study termination,
no PEC S 

### calculations are deemed necessary for this minor s oil metabolite.

### – 214 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.6 Predicted environmental concentration in grou ndwater (PEC GW 

### ) for the active

substance and relevant metabolites (OECD Annex IIIA , 9.6)

Reference: Predicted environmental concentrations of spirotetr amat and its main soil metabolites in groundwater recharge (PEC gw 

### ) based on calculations with FOCUS PEARL: Use in citrus in Europe Author(s), year: Röpke, B., 2006c Study/report number: M-277121-02-2, MEF-06/280, ame nded 12.06.2007 Guideline(s): Calculations – Not applicable GLP: Calculations – Not applicable Deviations: Calculations – Not applicable Validity: Study considered acceptable

and

Reference: Predicted environmental concentrations of spirotetr amat and its main soil metabolites in groundwater recharge (PEC gw 

### ) based on calculations with FOCUS PEARL: Use in leafy vegetable in Europe Author(s), year: Röpke, B., 2006d Study/report number: M-277164-02-2, MEF-06/385, ame nded 12.06.2007 Guideline(s): Calculations – Not applicable GLP: Calculations – Not applicable Deviations: Calculations – Not applicable Validity: Study considered acceptable

and

Reference: Predicted environmental concentrations of 4-methoxy cyclohexanone in groundwater recharge (PEC gw 

### ) based on calculations with FOCUS PEARL Author(s), year: Röpke, B., 2007b Study/report number: M-294996-01-1/MEF-07/477 Guideline(s): Calculations – Not applicable GLP: Calculations – Not applicable Deviations: Calculations – Not applicable Validity: Study considered acceptable

Material and methods: Predicted environmental concentrations of spirotetr amat and the metabolites spirotetramat-enol, spirotetramat-ketohydroxy, spirotetramat-MA-amide a nd 4-methoxy-cyclohexanone (the latter a major soil photolysis metabolite) in groundwater (PEC GW 

### , [μg L -1 

### ]) were calculated according to FOCUS guidance usin g FOCUS PEARL 3.3.3.

The degradation pathway of spirotetramat in soil wa s implemented as a multi-compartment model into FOCUS PEARL 3.3.3 taking into account a simple firs t order-reversible binding (SFO-RB) approach for th e metabolite spirotetramat-enol (Fig. B.8.6-1). Degra dation of the parent and all other compounds follow ed
SFO kinetics.

The major soil photolysis metabolite 4-methoxy-cycl ohexanone could not be adequately linked to the ove rall degradation model; therefore, 4-methoxy-cyclohexano ne was modelled for groundwater risk assessment on

### – 215 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

the basis of a pseudo application (taking into acco unt the maximum occurrence observed in the soil photolysis study and the molar ratio).

The SFO-RB model, used for spirotetramat-enol, is c onceptually similar to the kinetic sorption model implemented in FOCUS PEARL. The kinetic sorption mo del employed considers three phases, the dissolved phase, the equilibrium sorbed phase and the non-equ ilibrium sorbed phase (Figure B.8.6-2). The relatio n between dissolved and equilibrium sorbed phase is c haracterized by instantaneous equilibration between the concentrations in both phases and is described by t he Freundlich isotherm. The transfer between the equilibrium phase and the non-equilibrium sorbed ph ase is described by a first order (SFO) reaction wi th the corresponding sorption rate k d 

### . The ‘capacity’ of the non-equilibrium sorbed phas e is characterized by the parameter f NE 

### which is defined as the ratio between the Freundli ch coefficients in the equilibrium phase and the non-equilibrium sorbed phase. The degradation c onstant in the equilibrium domain k t 

### , which is used for the estimation of the first order pseudo half life, is valid in connection with the kinetic model, onl y. Kinetic sorption and degradation parameters stated in Table B.8.6-2, which are required for groundwater modell ing, were directly derived from the SFO-RB parameter (fo r more details refer to chapter ‘Aerobic degradatio n of relevant metabolites at 20 °C’, B.8.1.2.3).

The SFO-RB/kinetic adsorption approach was also sho wn to adequately describe the behaviour of spirotetramat-enol observed in the column leaching study.

Figure B.8.6-1: Multi-compartment model of spirotet ramat and metabolites used for groundwater modelling (taking into account a SFO-RB approach for spirotetramat-enol). Spirotetramat (SFO)

Spirotetramat-enol (mobile)

Spirotetramat-ketohydroxy (SFO)

Spirotetramat-MA-amide (SFO)

Sink 100 %

26 % 74 %

100 %

100 % Spirotetramat-enol (protected) SFORB

### – 216 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Figure B.8.6-2: Concept of the equilibrium and non- equilibrium domain of the soil system

(kinetic sorption model) implemented in FOCUS PEARL

.

Table B.8.6-1: Application data of spirotetramat ap plied as 150 OD formulation.

Crop Citrus Leafy vegetables a
Application rate [g ha -1 ] 288 b 72 Number of applications 2 2 Interval [days] 21 14
BBCH code 71 – 78 42 - 43 Crop interception [%] 70 c 70 d
Application dates: Châteaudun Hamburg Jokioinen Kremsmünster Piacenza Porto Sevilla Thiva -

-

-

- November 11 th , 28 th
November 11 th , 28 th
November 11 th , 28 th
November 11 th , 28 th Relative application date for all relevant scenarios (30 and 44 days after emergence)

a Leafy vegetables not defined in FOCUS groundwater, ‘ cabbage’ used as surrogate
b Maximum application rate assuming 96 g ai ha -1 m -1 tree height (maximum tree height 3 m)

c FOCUS default value for all stages of citrus (SANCO/32 1/2000 rev. 2)
d FOCUS default value for this crop and BBCH code (SANCO/3 21/2000 rev. 2)– 217 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.6-2: Input parameter for spirotetramat and metabolites (all input parameter not mentioned inthis table were set to default values).Compound Parameter ValueRemarksMolecular weight [g mol-1] 373.45Water solubility (20 °C, pH 7) [mg L-1] 29.9Vapour pressure (20 °C) [Pa] 5.6 × 10-9KFOC[L kg-1]281 Arithmetic mean, batch, n = 51/n0.94 Arithmetic mean, batch, n = 5DegT50soil [days]0.13 Geometric mean, lab, SFO, normalized, n = 4SpirotetramatPlant uptake factor0.5 FOCUS defaultMolecular weight [g mol-1] 301.38Water solubility (20 °C, pH 7) [mg L-1] 2700Vapour pressure (20 °C) [Pa] 3.64 × 10-10Unknown, estimate, US EPA (2000)KOC[L kg-1]55 Arithmetic mean, column leaching study, n = 41/n1.0 Unknown, worst caseDegT50soil [days]0.03aGeometric mean, lab (spirotetramat-enol degradationstudy), kinetic sorptionSFO according to the SFO-RB approach, normalized, n= 4Formation fraction [0 ... 1] (from parent) 1.00 Unknown, worst case assumptionkd[days-1]0.016Geometric mean, lab (spirotetramat-enol degradationstudy), SFO accordingto the SFO-RB approach, non-normalized, n = 4fNE[-]377bArithmetic mean, lab (spirotetramat-enol degradation study), ratio accordingto the SFO-RB approach, non-normalized, n = 4SpirotetramatenolPlant uptake factor0.5 FOCUS defaultMolecular weight [g mol-1] 317.38Water solubility (20 °C, pH 7) [mg L-1] 288Vapour pressure (20 °C) [Pa] 2.21 × 10-10Unknown, estimate, US EPA (2000)KFOC[L kg-1]63.7 Arithmetic mean, batch, n = 51/n0.92 Arithmetic mean, batch, n = 5DegT50[days]3.8Geometric mean, lab (spirotetramat-enol degradationstudy), SFO,normalized, n = 4Formation fraction [0 ... 1] (from spirotetramat-enol) 0.26 Arithmetic mean, lab (spirotetramat-enol degradation study), n = 4SpirotetramatketohydroxyPlant uptake factor0.5 FOCUS defaultMolecular weight [g mol-1] 335.40Water solubility (20 °C, pH 7) [mg L-1] 69.74Vapour pressure (20 °C) [Pa] 1.56 × 10-11Unknown, estimate, US EPA (2000)KFOC[L kg-1]9.3 Arithmetic mean, batch, n = 5SpirotetramatMA-amide1/n0.95 Arithmetic mean, batch, n = 5– 218 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Compound Parameter ValueRemarksDegT50[days]1.0Geometric mean, lab (spirotetramat-enol degradationstudy), SFO,normalized, n = 4Formation fraction [0 ... 1] (from spirotetramat-ketohydroxy) 1.00 Set to 1.00 during kinetic evaluationPlant uptake factor0.5 FOCUS defaultMolecular weight [g mol-1] 128.17Water solubility (20 °C, pH 7) [mg L-1] 53200 Unknown, estimate, US EPA (2000)Vapour pressure (20 °C) [Pa] 70.8 Unknown, estimate, US EPA (2000)KFOC[L kg-1]0 Unknown, worst case1/n0.9 Unknown, FOCUS defaultDegT50[days]0.6 Worst case, lab, SFO, non-normalized, n = 3Maximum occurrence [% of AR] 10.0 Soil photolysis4-methoxycyclohexanonePlant uptake factor0.5 FOCUS defaultaAs it is not possible to use a DT50value below 0.1 d in PEARL 3.3.3, a DT50of 0.1 days was used instead of the calculated value of 0.03 days, which represents a worst case forspirotetramat-enol inthis casebArithmetic mean value of 377 substituted with 99 (upper limit in PEARL 3.3.3)

### – 219 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Findings:

Table B.8.6-3: PEC GW 

### values [μg L -1 

### ] of spirotetramat and its soil metabolites from us e in citrus and leafy vegetables in Europe calculated with FOCUS PE ARL 3.3.3.

Crop Scenario Spirotetramat Spirotetramatenol Spirotetramatketohydroxy SpirotetramatMA-amide 4-methoxy

cyclohexanone

Piacenza < 0.001 < 0.001 < 0.001 < 0.001 < 0.001
Porto < 0.001 < 0.001 < 0.001 < 0.001 < 0.001
Sevilla < 0.001 < 0.001 < 0.001 < 0.001 < 0.001 Citrus

Thiva < 0.001 < 0.001 < 0.001 < 0.001 < 0.001

Châteaudun < 0.001 < 0.001 < 0.001 < 0.001 < 0.001 Hamburg < 0.001 < 0.001 < 0.001 < 0.001 < 0.001
Jokioinen < 0.001 < 0.001 < 0.001 < 0.001 < 0.001
Kremsmünster < 0.001 < 0.001 < 0.001 < 0.001 < 0.001
Porto < 0.001 < 0.001 < 0.001 < 0.001 < 0.001
Sevilla < 0.001 < 0.001 < 0.001 < 0.001 < 0.001 Leafy vegetables

Thiva < 0.001 < 0.001 < 0.001 < 0.001 < 0.001



### Conclusion: The leaching simulations resulted in PEC GW 

### values below 0.1 μg L -1 

### for spirotetramat and its soil metabolites
for all intended uses and all relevant FOCUS scenar ios. The risk for groundwater leaching is considere d low.

Comments (RMS):

• The geometric mean soil DT 50 

### of spirotetramat-ketohydroxy should be 3.9 days in stead of 3.8 days used for PEC calculations. However, this minor diff erence is not considered to have a significant impact onto the risk assessment of spirotetramat-ke tohydroxy. Therefore, the groundwater risk assessment is considered acceptable at all.

• It may be annotated, that degradation rates and fo rmation fractions of spirotetramat-ketohydroxy and spirotetramat-MA-amide obtained from the more simpl istic degradation schemes SFO  SFO  SFO  SFO, SFO  FOMC  SFO  SFO or SFO  DFOP  SFO  SFO (all lower tier approaches) are almost identical to the data obtain ed from the higher tier approach SFO  SFO-RB

 SFO  SFO, applied in the study above (also refer to cha pter ‘Aerobic degradation of relevant metabolites at 20 °C’, B.8.1.2.3). This is reasonab le since the release of spirotetramat-ketohydroxy from spirotetramat-enol as its precursor is almost exclusively attributed to the initial and fast degradation phase of the (mobile) spirotetramat-eno l fraction. The release of spirotetramatketohydroxy from the second and slower degradation phase of the strongly bound (immobile) spirotetramat-enol fraction is considerably lower. In this respect, a reasonable 1 st 

### tier groundwater
risk assessment may be solely based on obtained SFO degradation ra tes for all, the parent and all metabolites including spirotetramat-enol. The ‘rema ining’ strongly bound spirotetramat-enol fraction in the soil (representing the latter phase in the F OMC or DFOP kinetics model, which is per se not adequately addressed using SFO kinetics) is neither considered to be mobile (no potential for leaching as demonstrated in the column leaching stu dy) nor considered to significantly attribute to the further release of metabolites owing to its rat her slow degradation rate.

### – 220 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.7 Predicted environmental concentration in surf ace water (PEC SW 

### ) and sediment

(PEC SED 

### ) for the active substance and relevant metabolites (OECD Annex IIIA, 9.7, 9.8)

Reference: Predicted environmental concentrations of spirotetr amat and its main metabolites in surface water and sediment (PEC SW/SED 

### ) calculated according to FOCUS: Use in citrus in Europe Author(s), year: Röpke, B., 2006e Study/report number: M-277190-01-1, MEF-06/281 Guideline(s): Calculations – Not applicable GLP: Calculations – Not applicable Deviations: Calculations – Not applicable Validity: Study considered acceptable

and

Reference: Predicted environmental concentrations of spirotetr amat and its main metabolites in surface water and sediment (PEC SW/SED 

### ) calculated according to FOCUS: Use in leafy vegetables in Europe Author(s), year: Röpke, B., 2006f Study/report number: M-277176-01-1, MEF-06/386 Guideline(s): Calculations – Not applicable GLP: Calculations – Not applicable Deviations: Calculations – Not applicable Validity: Study considered acceptable

Material and methods:
Predicted environmental concentrations in surface w ater (PEC SW 

### ) and sediment (PEC SED 

### ) for spirotetramat, spirotetramat-enol, spirotetramat-ketohydroxy, 4-me thoxy-cylcohexanone and 4-methoxy-cyclohexylaminocarboxylic acid (the latter two major in water photolysis and, partly, in soil photolysis) were c alculated according to FOCUS guidance based on FOCUS surface water tools STEPS 1 – 4 (for metabolites STEPS 1 and 2, only).

At FOCUS surface water STEP 4 (for use in citrus on ly), mitigation of drift deposition was achieved by introducing buffer zones (vegetation filter strips) . Reduced drift deposition based on a 5-m-buffer zo ne (Table B.8.7-1) was calculated using the FOCUS drift calcu lator. No mitigation of runoff or drainage was assu med.

### – 221 –

Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Table B.8.7-1: Application data of spirotetramat ap plied as 150 OD formulation.

Crop Citrus Leafy vegetables Application rate [g ha -1 ] 288 a 72 Number of applications 2 2 Interval [days] 21 14
BBCH code 71 – 78 42 - 43 Crop interception [%] FOCUS STEP 1 FOCUS STEP 2 FOCUS STEP 3 & 4 None 70 b (full canopy) Calculated by the model None 70 c (full canopy) Calculated by the model Drift deposition [%] FOCUS STEP 1, 2 & 3 FOCUS STEP 4 (5 m buffer zone) Ditch Stream Default values

7.51 9.01 e Default values Not calculated

Region/Scenario: FOCUS STEP 1 FOCUS STEP 2 FOCUS STEP 3 & 4 Not defined South EU D6 (Thiva), R4 (Roujan) Not defined South EU, North EU D3 (Vredepeel, 1 st ), D4 (Skousbo), D6 (Thiva), R1 (Weiherbach, 1 st ), R2 (Porto, 1 st ), R3 (Bologna, 1 st ), R4 (Roujan, 1 st ) Application timing/window: FOCUS STEP 1 FOCUS STEP 2 FOCUS STEP 3 & 4 Not defined October to February Start: D6: November 1 st
R4: November 1 st Appl. window: 30 + 21 days Appl. date calculated by PAT d Not defined March to May f Start: D3: May 13 th D4: July 20 th D6: September 23 rd R1: May 8 th R2: April 23 rd

R3: March 25 th

R4: March 25 th Appl. window: 30 + 14 days Appl. date calculated by PAT d
RunOff/Drainage: FOCUS STEP 1 & 2 FOCUS STEP 3 FOCUS STEP 4 Default values Modelled by PRZM and MACRO Modelled by PRZM and MACRO, no runoff/drainage mitigation

assumed Default values Modelled by PRZM and MACRO

Not calculated

a Maximum application rate assuming 96 g ai ha -1 m -1 tree height (maximum tree height 3 m)
b FOCUS default value for all stages of citrus (SANCO/48 02/2001-rev.2)

c FOCUS default value for this crop and BBCH code (SANCO/4 802/2001-rev.2)
d PAT denotes pesticide application timer

e Number stated corrected for additional indirect dri ft input from the upstream catchment (i.e. 20 %)

f In respect to runoff /drainage, the application ti ming ‘March to May’ is worst case for South EU, for North EU this application timing is representative for the indented use– 222 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Table B.8.7-2: Input parameter for spirotetramat and metabolites (input parameter not mentioned in this table were set to default values).ValueCompound ParameterSTEP 1 & 2 STEP 3 & 4RemarksMolecular weight [g mol-1] 373.45Water solubility (20 °C, pH 7) [mg L-1] 29.9Vapour pressure (20 °C) [Pa] - 5.6 × 10-9KFOC[L kg-1]281 Arithmetic mean, batch, n = 51/n- 0.94 Arithmetic mean, batch, n = 5DegT50soil [days] 0.13 Geometric mean, lab, SFO, normalized, n = 4DegT50water [days] 0.78 Geometric mean, entire system, SFO, n = 2DegT50sediment [days] 0.78 1000STEP 1 & 2: Geometric mean, entire system, SFO, n =2STEP 3 & 4: Worst case default valueSpirotetramatDegT50entire system [days] 0.78 - Geometric mean, entiresystem, SFO, n = 2Molecular weight [g mol-1] 301.38Water solubility (20 °C, pH 7) [mg L-1] 2700KOC[L kg-1] 55 Arithmetic mean, column leaching study, n = 4DegT50soil [days] 1.16Geometric mean, lab (spirotetramat-enol degradationstudy), SFO recalculated fromFOMC-DT90divided by 3.32, normalized, n = 4DegT50water [days] 47.3 Geometric mean, entire system, SFO, n = 2DegT50sediment [days] 47.3 Geometric mean, entire system, SFO, n = 2DegT50entire system [days] 47.3 Geometric mean, entire system, SFO, n = 2Maximum occurrence soil [%] 100.0 Not known, worstcase assumptionSpirotetramatenolMaximum occurrence entire system [%] 97.5NotperformedMolecular weight [g mol-1] 317.38Water solubility (20 °C, pH 7) [mg L-1] 288KFOC[L kg-1] 63.7 Arithmetic mean, batch, n = 5DegT50[days] 3.8 Geometric mean, lab (spirotetramat-enoldegradation study), SFO, normalized, n = 4DegT50water [days] 1000 No degradation observed, worst case default valueDegT50sediment [days] 1000 No degradation observed, worst case default valueDegT50entire system [days] 1000 No degradation observed,worst case default valueMaximum occurrence soil [%] 24.0SpirotetramatketohydroxyMaximum occurrence entire system [%] 40.5NotperformedMolecular weight [g mol-1] 128.17Water solubility [mg L-1] 53200 Unknown, estimate, US EPA, 2000aKOC[L kg-1] 2.6 Unknown, estimate, US EPA, 2000a4-MethoxycyclohexanoneDegT50[days] 0.6NotperformedWorst case, lab, SFO, non-normalized, n = 3– 223 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008ValueCompound ParameterSTEP 1 & 2 STEP 3 & 4RemarksDegT50water [days] 1000 No degradation observed, worst case default valueDegT50sediment [days] 1000 No degradation observed, worst case default valueDegT50entire system [days] 1000 No degradation observed,worst case default valueMaximum occurrence soil [%] 10.0aSoil photolysis studyMaximum occurrence entire system [%] 17.5 Phototransformation study in natural waterMolecular weight [g mol-1] 173.21Water solubility [mg L-1] 26570 Unknown, estimate, US EPA, 2000aKOC[L kg-1] 10 Unknown, estimate, US EPA, 2000aDegT50[days] 1000 Unknown, not observed in soil, value does not effect the calculationDegT50water [days] 1000 No degradation observed, worst case default valueDegT50sediment [days] 1000 No degradation observed, worst case default valueDegT50entire system [days] 1000 No degradation observed,worst case default valueMaximum occurrence soil [%] 0.0bNot observed in soil4-MethoxycyclohexylaminocarboxylicacidMaximum occurrence entire system [%] 11.3NotperformedPhototransformation study in natural wateraSet to 0.0 % of AR in the original study, re-calculated by the RMS based on 10.0 % of AR. See comments.bAt FOCUS STEP 1 & 2 set to 1 × 10-10(zero value not accepted).– 224 –Spirotetramat – Volume 3, Annex B.8, Fate and behaviour in the environmentApril 2008Findings:Table B.8.7-3: Predicted environmental concentrations of spirotetramat and metabolites in surface water (PECSW, [μg L-1]) and sediment (PECSED,[μg kg-1]) after use in citrusfollowing FOCUS surface water STEPS 1 - 4.FOCUS surface water STEP 1 STEP 2 STEP 3 STEP 4 (5-m-buffer zone)CompartmentRegion/Scenario - South EU D6 - Ditch R4 - Stream D6 - Ditch R4 - StreamSpirotetramat 84.9 11.6 8.43 6.51 5.86 ncSpirotetramat-enol 168 15.6 nc nc nc ncSpirotetramat-ketohydroxy 46.5 8.59 nc nc nc ncMethoxy cyclohexanone 8.38 1.39 nc nc nc ncSurface water(PECSW)[μg L-1]Methoxy cyclohexylaminocarboxylic acid 1.58 1.21 ncnc nc ncSpirotetramat 196 6.53 4.16 0.753 2.92 ncSpirotetramat-enol 90.2 8.00 nc nc nc ncSpirotetramat-ketohydroxy 29.1 5.34 nc nc nc ncMethoxy cyclohexanone 0.218 0.036 nc nc nc ncSediment(PECSED)[μg kg-1]Methoxy cyclohexylaminocarboxylic acid 0.156 0.119nc nc nc ncnc denotes not calculatedTable B.8.7-4: Predicted environmental concentrations of spirotetramat and metabolites in surface water (PECSW, [μg L-1]) and sediment (PECSED,[μg kg-1]) after use in leafy vegetablefollowing FOCUS surface water STEPS 1 - 3.FOCUS surface water STEP 1 STEP 2STEP 3CompartmentRegion/Scenario -NorthEUaSouthEUD3ditchD4pondD4streamD6ditchR1pondR1streamR2streamR3streamR4streamSpirotetramat 18.1 0.585 0.585 0.399 0.0129 0.324 0.398 0.0129 0.261 0.350 0.368 0.261Spirotetramat-enol 37.1 0.850 0.950 nc nc nc nc ncnc nc nc ncSpirotetramat-ketohydroxy 9.48 0.519 0.662 nc nc ncnc nc nc nc nc ncMethoxy cyclohexanone 1.72 0.071 0.071 nc nc nc ncnc nc nc nc ncSurface water(PECSW)[μg L-1]Methoxy cyclohexylaminocarboxylic acid 0.069 0.0610.061 nc nc nc nc nc nc nc nc ncSpirotetramat 49.1 0.328 0.328 0.119 0.007 0.021 0.108 0.006 0.033 0.026 0.071 0.034Spirotetramat-enol 20.1 0.452 0.505 nc nc nc nc ncnc nc nc ncSpirotetramat-ketohydroxy 6.01 0.324 0.415 nc nc ncnc nc nc nc nc ncMethoxy cyclohexanone 0.043 0.002 0.002 nc nc nc ncnc nc nc nc ncSediment(PECSED)[μg kg-1]Methoxy cyclohexylaminocarboxylic acid 0.007 0.0060.006 nc nc nc nc nc nc nc nc ncnc denotes not calculatedaCalculated by the RMS

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

Conclusion:
Maximum PEC SW 

### values of spirotetramat in surface water after use in citrus were calculated to be 84.9, 11.6, 8.43 and 5.86 μg L -1 

### at FOCUS surface water STEP 1, 2, 3 and 4 (STEP 4 considering drift mitigation only, buffer zone 5 m). Following application in leafy ve getables, maximum PEC SW 

### of spirotetramat were calculated to be 18.1, 0.59 and 0.40 μg L -1 

### at STEP 1, 2 and 3 (no STEP 4 calculation conducte d).

Maximum PEC SW 

### for the metabolites spirotetramat-enol, spirotetra mat-ketohydroxy, spirotetramat-MAamide, 4-methoxy-cylcohexanone and 4-methoxy-cycloh exylaminocarboxylic acid were calculated to be 15.6, 8.59, 1.39 and 1.21 μg L -1 

### , respectively, at FOCUS surface water STEP 2 after use in citrus and 0.95, 0.66, 0.071 and 0.061 μg L -1 

### , respectively, after use in leafy vegetables.

In sediment, maximum PEC SED 

### values of spirotetramat after use in citrus were c alculated to be 196, 6.53, 4.16 and 2.92 μg kg -1 

### at FOCUS surface water STEP 1, 2, 3 and 4 (STEP 4 considering drift mitigation only, buffer zone 5 m). Following application in leafy ve getables, maximum PEC SW 

### of spirotetramat were calculated to be 49.1, 0.33 and 0.12 μg kg -1 

### at STEP 1, 2 and 3 (no STEP 4 calculation conducte d).

Maximum PEC SED 

### values for the metabolites spirotetramat-enol, spi rotetramat-ketohydroxy, 4-methoxycylcohexanone and 4-methoxy-cyclohexyl-aminocarboxy lic acid were calculated to be 8.00, 5.34, 0.036 an d 0.12 μg kg -1 

### , respectively, at FOCUS surface water STEP 2 after use in citrus and 0.51, 0.42, 0.002 and 0.006 μg kg -1 

### , respectively, after use in leafy vegetables.

Comments (RMS):

• The geometric mean soil DT 50 

### of spirotetramat-ketohydroxy should be 3.9 days in stead of 3.8 days used for PEC calculations. However, this minor diff erence is not considered to have a significant impact onto the risk assessment of spirotetramat-ke tohydroxy. Therefore, the surface water and sediment risk assessment is considered acceptable a t all.

• In the original study report, maximum occurrence o f 4-methoxy-cyclohexanone in soil was set to 0.0 % of AR. However, since 4-methoxy-cyclohexanone is major in the soil photolysis (10.0 % of AR), the PEC SW/SED 

### were re-calculated by the RMS.

• In the original study report, no results on applic ation on leafy vegetables in North-EU at FOCUS surface water STEP 2 were presented. These data wer e calculated and added by the RMS.

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Spirotetramat – Volume 3, Annex B.8, Fate and behav iour in the environment April 2008

B.8.9 Predicted environmental concentration in the air (PEC A 

### ) (OECD Annex IIIA, 9.9)

Based on the low vapour pressure of spirotetramat a nd spirotetramat-enol (< 10 -9 

### Pa) no significant transfer into the atmosphere is expected to occur. In the ev entuality that spirotetramat or spirotetramat-enol enters the atmosphere, both compounds are expected to rapi dly degradate owing to reaction with OH radicals (atmospheric half-life < 2 hrs based on Atkinson ca lculation).