DP Barcode: D425260 MRID: 495 35601

1 Reference

M iller, A ., Bowers, L., Dyer, D., and Jerkins, E. (2014). Determination of the Residues of Imidacloprid

and its Metabolites 5

- Hydroxy Imidacloprid and Imidacloprid Olefin in Bee Relevant Matrices
Collected from Cherry Trees following Foliar Appl ication of Imidacloprid over Two Successive Years

. Study ID EBNTY008

. Bayer Crop Science, Research Triangle Park, NC. MRID 49535601



### 1. STUDY INFORMATION

Chemical: Imidacloprid PC Code 129099

Test Material Ga ucho 600 FL Admire Pro

Systemic Protectant ( SC) Purity 4 2.9

– 43.5 % a.i. w.w.

Study Type: Non

- Guideline field residue study on c herry to establish imidacloprid and

metabolite levels in pollen, nectar , flowers, leaves and soil after two successive
annual foliar applications.

Sponsor: Bayer Cro p Science AG

2 T. W. Alexander Drive
Research Triangle Park, NC

USA 27709 Performing Laboratories:
( Analytical ; Pollen, Nectar ): Bayer Crop Science
AG, 2 T. W. Alexander Drive Research Triangle

Park, NC USA 27709
( Analytical; Leaf, Flower, Soil): SynTech Research
Laboratory Services, 17745 S. Metcalf Ave,

Stillwell, KS 66085
( Trial NT00 7 / NT008

- 12ZA ): Reality Research

, 5916 South Ave, Williamson, NY 14589
(Trial NT0 16 / NT017

- 12ZA ): Columbia Ag

Research Inc. 5601 Binns Hill Dr., Hood River, OR

97031 ) Sp onsor Study

ID: EBNTY008
Study
Completion

Date: 12 December 2014

Experiment Start/End Date: 12 Aug. 2012

– 3 Sept. 2014
Study Location: 4 Field Trials:

Alton, NY (NT00 7 ) ; Lyons, NY (NT00 8 ) ; Mosier, OR (NT016 & NT017)

GLP Status: GLP

- complia nt ; protocol reviewed by EPA, PMRA, CDPR



### 2. REVIEWER INFORMATION

Primary Reviewer:

Signature: Keith Sappington, M.S., Senior Biologist/Science Advisor, ERB5,

Environmental Fate and Effects Division, Office of Pesticide Programs, USEPA

Date: 1 / 28 / 2016



### 3. EXECUTIVE SUMMARY

This study was designed to measure the magnitude of imidacloprid residues in bee

- relevant pollen and
nectar samples and in/on flowers, leaves, and soil from cherry trees after five post bloom foliar

applications of 0. 1 lb a.i./ A conducte d the previous growing season (current U.S. label rate). Four separate

trials were conducted in NY (NT007, NT008) and OR (NT016, NT017) cherry orchards , each with a treated

(TRTD) and untreated control (UTC) plot. Each year, five foliar applications (0. 1 lb a.i./ A x 5 ) were made,

with a retreatment interval ranged from 8

- 1 1 days. The first year applications (2012) were made after

cherry harvest in late summer/early fall (BBCH 91; shoot growth completed, green foliage). The second

year applications were ma de prior to cherry harvest during June/July 2013 (BBCH stage 71

- 73). SamplesDP Barcode: D425260 MRID: 495 35601

2 of flowers, pollen, nectar and leaves were made at various sampling times during the flowering period of

the growing season after pesticide application. Soil samples were also co llected. QA/QC included field
and lab storage/stability samples, blanks, and matrix spikes to validate analyte recovery.

Results indicate post

- bloom, foliar application of approximately 0. 1 lb a.i./A x 5 result s in residues of total
IMI (sum of IMI, IMI

- olefin and 5

- OH

- IMI) in individual composite nectar samples of up to 10 ppb and

100 0 ppb in pollen

. Mean residues of total IMI in nectar varied from 1.6

– 3.7 ppb and th ose for pollen

varied from 1.7

- 422 ppb

. Within each trial /year , total IMI residues i n pollen were approximately 1

- 2
orders of magnitude greater than those observed in nectar . Mean concentrations of total IMI in cherry

nectar measured after foliar applications during the previous late summer/early fall (Year 1) were about

2X those measured after foliar applications during late spring/early summer (year 2) across all trials. A
larger difference was observed in mean pollen concentrations (about 10X) across trial years. It is not clear
if this finding reflects seasonal differences in IMI tra nslocation or longer intervals between pesticide
application and residue measurement associated with year 2.

Mean concentrations of total IMI measured in pollen from the NY study trails were about 10X those
measured in pollen from the OR trial sites. Simi lar results were observed with total IMI residues measured

in flowers and leaves. T wo metabolites of IMI (IMI

- olefin and 5

- OH

- IMI) typically represented < 10% of
total IMI residues in pollen and nectar. The predictability of total IMI residues in nectar and pollen based
on measured concentrations in other plant matrices (flowers, leaves) was generally poor. Limited
sampling of plant matrices over each growing season prevented determination of reliable estimates of
dissipation kinetics (DT 50 ) in plant mat rices.



### 4. STUDY VALIDITY

Guideline Followed: Non

- guideline study (protocol was reviewed by EPA/PMRA/CDPR)

Guideline Deviations: N/A

Other Deviations: S ample times to freezer storage sometimes exceeded the 4 hours

specified in the protocol, but samples w ere maintained in a cooler with

blue ice, so this deviation does not impact the study results.

Classification: ACCEPTABLE

Rationale: No major deviations were identified in this study that would affect the

scientific integrity of this study
Reparability : N/A



### 5. MATERIALS AND METHODS

Test Material Characterization

Test item: Ga ucho 600 FL Admire Pro

Systemic Protectant SC
(Imidacloprid) 550 g a.i./L CAS #: 138261

- 41

- 3

Description: N/A Purity: 42.9 %. & 43.5% w/w

Lot No./Batch No. Batch No. NK41BX01 70;

NK41CX0578 Density: 1.41

– 1.54 g/mL

Material Source: Bayer Crop Science Cert. # 212PG1962; 213CJ2446

Material Receipt

Date: Not reported Analysis

Date: 03/30/2012 ; 12/03/2012

Expiration Date: 03/31/2014 ; 12/3/2014 Solubility: 0.51

- 0.61 g/LDP Barcode: D425260 MRID: 495 35601

3 Test Material Characterization

Storag e of Test Mat’l : Ambient ( 47

- 8 3 o F) Sample

Storage:

- 1 3 o to

- 1 8 o F

5A. S TUDY D ESIGN

This study was designed to measure the magnitude of imidacloprid residues in bee

- relevant pollen and nectar samples and in/on flowers, leaves, and soil from cherry trees following five pre

- bloom foliar
applications per year of ADMIRE PRO Systemic Protectant containing 550 g/L imidacloprid (IMI)

. In
addition to quantifying the magnitude of residues, this study was designed to evaluate the following

additional attributes:
1) The extent of year

- to

- year ‘ carry over ’ of pesticide residues measured in plant and soil

matrices
2) The dissipation rates of IMI /metabolite residues over time in plant matrices
3) The extent to which toxicological relevant metabolites (IMI

- olefin, 5

- hydroxy

- IMI ) occurred in

these matrices
4) The relationship of IMI/metabolite residues among different plant tissues (pollen, nectar, leaf,

flower).

Four separate trials were conducted , each with a treated (TRTD) and untreated control (UTC) plot. Each

treated plot wa s divided into 5 subplots each with at least 3 trees (15 total trees). Each year, five foliar

applications (0. 1 lb a.i./ A x 5 ) were made at intervals ranging from 8

- 11 days. The first year applications

(2012) were made after cherry harvest in late summe r/early fall (BBCH 91; shoot growth completed,

green foliage). The second year applications were made prior to cherry harvest during June/July 2013

(BBCH stage 71

- 73). Five composite samples of flowers, pollen, nectar and leaves were made at various

samp ling times during the flowering period from each TRTD plot/year . Nine composite soil samples were collected prior to treatment and at the end of the growing season from each TRTD plot/year. Control

plots were sampled for all cherry matrices (two composit es) and soil (9 composites) at the same time the

TRTD plots were sampled. QA/QC included field and lab storage/stability samples, blanks, and matrix

spikes to validate analyte recovery.

5 B

. A PPLICATION T IMING AND R ATES

During each of the two successive years, five foliar applications of approximately 0. 1 lb a.i./A were applied

at intervals from 8

- 11 days apart at each of the four treatment sites (total annual application rate = 0.5 lb
a.i./A). All applications were made with adjuvant (Dyne

- Amic, 0. 3 % ) us ing a ground

- based air blast

equipment

. The volume of application was similar across trials and years (data not shown). In addition, a

control (untreated) site was also included for each of the four trials. A ll FIRST YEAR applications were

made after che rry harvest at BBCH growth stages 9 1 (shoot growth completed, foliage still fully green).

All YEAR TWO applications were initiated prior to cherry harvest bet ween BBCH growth stages 73 and 75
(second fruit fall

– fruit about half size).

Inf ormation on application timing is shown in the T able 1

.DP Barcode: D425260 MRID: 495 35601

4 Table 1 . Summary of foliar application rates and timing

Location Cherry

Variety Year Appl. No. Appl. Date (DAFA/Stage) 1 Individual Rate (lb a.i./A) Total Rate

(lb a.i./A)

Trial NT00 7

- 12ZA

Alton, NY Mont

morency 2012 1

- 5 8/31/2012

– 10/3/2012

( 0

- 33/ BBCH 9 1) 0. 100 0.50

2013 1

- 5 6/5// 2013

- 7/15/2013

( 278

- 318/ BBCH 7 3

- 85) 0.100

- 0.102 0.50

Trial NT00 8

- 12ZA

Lyons, NY Mont

morency 2012 1

- 5 9/3/2012

– 10/8/2012

(0

- 35 / BBCH 91 ) 0.10 0 0.5 0

2013 1

- 5 6/5/2013

– 7/15/2013

( 275

- 315 / BBCH 74

- 85 ) 0.100 0.5 0

Trial NT0 1 6

- 12ZA

Mosier, OR Skeena 2012 1

- 5 8/11/2012

– 9/13/2012

( 0

- 33/ BBCH 9 1) 0.098

- 0.102 0.5 0

2013 1

- 5 6/5/2013

– 7/11/2013

( 298

- 334 / BBCH 74

- 87 ) 0.099

- 0.102 0.5 0

Trial N T01 7

- 12ZA

Mosier, OR Bing 2012 1

- 5 8/11/2012

– 9/13/2012

( 0

- 33/ BBCH 9 1) 0.101

- 0.102 0.5 1

2013 1

- 5 5/28/2013

– 7/1/2013

( 290

- 324 / BBCH 75

- 87 ) 0.100

- 0.102 0.5 0

1 DAFA = days after 1 st application; BBCH = plant growth stage

.

5C. S TUDY S ITE L OCATION AN D C HARACTERISTICS

Various aspects of the study sites are summarized in Table 2. In general, soil type, plot size, row /plant

spacing , and tree age wer e similar among th e three study sites/trials. Three different variet ies of cherry
were used among the f our trials

.

Table 2 . Summary of Che r ry study site characteristics (treated sites only)

. Attribute Trial NT007

- 12ZA

Alton, NY Trial NT008

- 12ZA

Lyons, NY Trial NT016

- 12ZA

Mosier, OR Trial NT017

- 12ZA

Mosier, OR

Plot ID TRTD 12/13 TR TD12/13 TRTD12/13 TRTD12/13

Variety Mont morency Montmorency Skeema Bing

Crop Age 12 15 15 1 2

Soil Type Sand y loam Sand y loam Sand y loam Sand y loam

Plot Length 176 ft 272 ft 20 4 ft 170 ft

Plot Width 45 ft 60 ft 5 4 ft 60 ft

Plot Area 7920 ft 2 16320 ft 2 11016 ft 2 2 0,200 ft 2

Row Spacing 1 5 ft 2 0 ft 18 ft 2 0 ft

Plant

Spacing 8 ft 1 6 ft 12ft 1 0 ftDP Barcode: D425260 MRID: 495 35601

5 Attribute Trial NT007

- 12ZA

Alton, NY Trial NT008

- 12ZA

Lyons, NY Trial NT016

- 12ZA

Mosier, OR Trial NT017

- 12ZA

Mosier, OR

within Row

Lat . Long. N N 43°10.568'

W76°57.674 N43°09.733'

W76 ° 58.048 N4 5 ° 40

. 617 '

W121°22.094 N4 5 ° 40

. 329 '

W121°2 1

. 973

A summary of soil and meteorological dat a from the three study sites is shown in Table 3. Soil from trial
NT007 contained the greatest amount of organic carbon (3.4%) while soils from the other trials ranged

from 0.9 to 1.5% organic carbon . Soils from all four sites had similar % sand (53%

- 67% ) but the % silt at
site NT007 (17%) was about half that of the other sites (27

- 33%). Total p recipitation was similar among
sites, although year 2 data were unavailable from Trial NT017 because trees were removed by the grower
in July 2013 from this site.

The two NY trial sites (NT007 and NT008) were in close proximity as were the two OR sites (NT016 and

NT017; see Appendix A ). Therefore, the NY sites shared the same weather data as did the OR sites
( Figure 1 ). Therefore, to the extent that variation in weather affects residue levels in cherry trees, the NY
sites and OR sites can be considered replicates from a weather perspective. The NY trial sites were not
irrigated during the study period while the OR sites were irrigated (2

- 6 in/month from May

- Se pt of each
year).

Table 3 . Soil and meteorological characteristics of the study sites *

* (from Table 3 of study report)DP Barcode: D425260 MRID: 495 35601

6 F igure 1

. Monthly precipitation for the 4 study trial sites (NT007 & NT008 share the same weather data
and NT016 and NT017 also share the same data for year 1 only )

5D. S AMPLE C OLLECTION
, H ANDLING
, P ROCESSING

Plant Matrices. Samples that were treated in 2012 were harvested in 2013 (this period inclusively
considered year 1 of the st udy), and samples treated in 2013 were harvested in 2014 (year 2 of the study)

. Each treated plot TRTD was divided into 5 subplots. In trials NT007

- 12ZA, NT008

- 12ZA, and NT016

- 12ZA,
one composite (separate runs through each subplot) samples of cherry flowe rs for direct analysis, flowers
to be processed for nectar and pollen, and leaves was collected by hand at each sampling period from
each subplot (for 5 total samples) in both study years. In trial NT017

- 12ZA, no samples after the fall 2013
applications we re collected because the trees were removed from the orchard by the grower. Samples
were taken from the same trees at each sampling interval and in successive years. Cherry pollen and
nectar samples were harvested by hand at the field trial site from the f lowers collected for processing.

Cherry flowers were collected when the trees were at growth stages of BBCH 61 and 65 (BBCH 61:
beginning of flowering, about 10% of flowers open; BBCH 65: full flowering, at least 50% of flowers open
and first petals fall ing), corresponding to 205 to 218 days after the last application (DAA) in year 1 and 274

to 303 DAA in year 2. The exception is trial NT007

- 12ZA, which could not harvest BBCH 61 flowers in year

2 due to weather. Cherry leaves were collected when the trees were at growth stages of BBCH 65 and 69
(BBCH 69: end of flowering, all petals fallen), corresponding to 209 to 232 DAA in year 1 and 279 to 312

DAA in year 2

.

Table 4 . Sampling dates and cherry developmental stages for plant re sidue analysis

Pollen, Nectar and Flower Sampling Leaf Sampling

Year 1 Year 2 Year 1 Year 2

BBCH* DAA* BBCH DAA BBCH DAA BBCH DAA

NT007

61 212 61 302 65 215 65 302

65 215

--- --- 69 232 69 312

NT00 8

61 208 61 300 65 210 65 303

65 210 65 303 69 227 69 312DP Barcode: D425260 MRID: 495 35601

7 Pollen, Nectar and Flower Sampling Leaf Sampling

Year 1 Year 2 Year 1 Year 2

BBCH* DAA* BBCH DAA BBCH DAA BBCH DAA

NT016

61 214 61 274 65 218 65 279

65 218 65 279 69 228 69 285

NT017

61 205

--- --- 65 209

--- ---

65 209

--- --- 69 223

--- --- *BBCH = B iologische Bundesanstalt, B undessortenamt und CH emische Industrie growth stage scale for cherry ; DALA

= d ays after last application

Soil. Nine soil samples were collected twice during the first year of the study, prior to the first a.i.
application in 2012 and after the last sampling per plot in 2013, with the exception of trials NT007

- 12ZA

and NT008

- 12ZA, i n which the last leaf collection took place after the year 1 2013 soil collection. Soil
samples were also collected twice during the second year of the study, after the last application in 2013
and after the last sampling in 2014, with the exception of tri al NT017

- 12ZA, in which the 2014 samples

were not collected due to the tree removal. Soil samples were collected from the surface (6 Inches or 15
cm deep) of the plots using a soil core sampler and placed In a plastic bag. In each sampling period,
sample l ocations were randomly selected from the inner half of the plot (e.g. the inner 100 ft X 200 ft [30
X 60 m] area of a 200 ft x 400 ft [60 x 120 m] plot)

.

Control Plots. Two composite samples of all c herry matrices and nine composite samples of soil were
collected from the control plot of each trial at the same sampling periods as for the treated samples of
their sample type.

Sample Storage

. All samples were protected from sunlight and placed in field coolers containing ice

substitute and the transit sta bility samples. Upon arrival at the site facility, all composite samples were
placed directly into frozen storage except the flowers to be processed for nectar and pollen, which were
processed for pollen and nectar by the field investigators

.

The flower s to be processed for nectar and pollen were processed within 3 days of collection. Processing
delays occurred because the flowers had to be dried for several hours to allow pollen collection to occur

and the prin cipal field investigators were under time c onstraints because blooming occurred
simultaneously across multiple trials. To process the flowers, first nectar from the floral nectary was
removed using a micropipette and placed into a glass collection vial. The flowers were left out to dry, and
then po llen was removed from the anthers of the same flower samples by vacuum aspiration with filter
tips. All resulting nectar and pollen samples were labeled and placed in the freezer immediately after they
were generated. In all trials, when sample times to f reezer storage exceeded the 4 hours specified in the
protocol, the samples were kept cool in a cooler with blue ice

. Composite samples were placed into
labeled (study number and sample number) containers for shipment. Samples remained frozen until

receipt at the Bayer CropScience facility (Stilwell, KS or RTP, NC).

5E. A NALYTICAL M ETHODS

The residues of imidacloprid, 5

- hydroxy imidacloprid, and imidacloprid olefin) were quantitated by high
performance liquid chromatography/triple stage quadrupole mass spe ctrometry (LC/MS/MS) and LC/highDP Barcode: D425260 MRID: 495 35601

8 resolution mass spectrometry (LC/HRMS) using stable isotopically labeled internal standards. Details of
the analytical methods are provided in the study report.

The limit of detection (LOD) for the total residue is the hi ghest LO D value for an individual analyte in a
particular matrix. The limit of quantification (LOQ) for the total residue is the highest LOQ value for an
individual analyte in a particular matrix. The LOD and LOQ values for each matrix are shown below in

T able 5

.

Table 5 . Limits of quantification and detection for imidacloprid and metabolites

5F. Q UALITY A SSURANCE

Storage Stability

. Storage stability studies and transit spikes indicate that the imidacloprid residues would

have been stable during frozen storage for at least 1080 days (36 months) in cherry flower and leaf
matrices and for at least 1281 days (42 months) in soil matrices prior to analysis

.

Transit Stability. To ensure that imidacloprid and its metabolites 5

- hydroxy imidacloprid and imidacloprid
olefin were stable in nectar and pollen, sample sets of two pollen unfortified controls, four fortified pollen
samples, two nectar surrogate controls, and four fortified nectar surrogate samples, were prepared yearly

at Bayer CropScience and shipped frozen via FedEx to each test site. The transit stability samples, fortified
with approximately 0.10 ppm each of imidacloprid and its metabolites 5

- hydroxy imidacloprid and
imidacloprid olefin, accompanied the respective study samp le types throughout the sampling, shipping,

and storage events of the field phase of the study. They were analyzed after all sample analyses of a given
matrix and served to demonstrate stability of imidacloprid residues through the storage intervals and

tr ansport conditions encountered by the samples.

Spike Recovery. The method for determining imidacloprid, 5

- hydroxy imidacioprid, and i midacloprid

olefin residues in/on cherry and soil samples was validated by measuring the recovery of these analytes

from control matrices fortified at their respective matrix LOQs. A dditional recoveries at higherDP Barcode: D425260 MRID: 495 35601

9 fortification levels validated the method for the highest residues observed in individual matrices.
Concurrent recoveries of imidacloprid, 5

- hydroxy imidacloprid, and imidacloprid olefin from fortified
samples were measured with each set of samples to verify method performance

.



### 6. RESULTS:

6.A. Q UALITY A SSURANCE R ESULTS

Sample Storage. The maximum storage period of frozen samples in this study for imidacloprid w as 452
days for cherry flowers, 462 days for cherry leaves, 129 days for cherry nectar, 500 days for cherry pollen,
and 668 days for soil prior to extraction. These storage times are within the bounds of the storage
stability results described earlier and indicate that storage times are acceptable.

Transit Stability. The transit stability analyses indicate that the imidacloprid residues are stable (<30%
decomposition) during concurrent freezer storage with the samples from this study. Mean transit

recover ies for IMI, IMI

- Olefin, and 5

- hydroxy IMI ran ge from 91%

- 96% in nectar and 88 %

- 108 % in pollen.

Matrix Spike Recoveries. As indicated in Table 5 of the study report, all matrix spike recoveries (poll en, nectar, flower, leaf, soil), t he overall mean of the recoveries for each matrix was within the acceptable
range of 70 to 120%, and the standard deviation values were below 20%.

6. B

. M AGNITUDE OF R ESIDUES IN B EE

- R ELEVANT M ATRICES

Nectar. Summary statistics of t he overall magnitude of IMI and related me tabolites in c herry nectar is

shown in Table 6

. These statistics reflect analysis of individual composite samples (usually 5 replicates per
sampling date) among the sampling times within each trial. Since IMI

- olefin and 5

- OH IMI are considered

of similar toxicity as parent IMI, this discussion focuses on total IMI residues (sum of IMI, IMI

- olefin and 5

- OH

- IMI). Across all four trials, m ean residues of total IMI in cherry nectar range from approximately 1.6

–
3.7 ppb while maxima range from 3.4

– 10 pp b

.

Table 6 . Magnitude of IMI and related metabolites in c herry nectar (ppb).

Trial NT0 07 (n=15)* Trial NT008 (n=2 0) *

Olefin 5

- OH IMI Total Olefin 5

- OH IMI Total

Mean 0.45 1.9 1.3 3.7 0.41 0.55 1.6 2.5

SD 0.35 1.4 0.8 2.2 0.26 0. 38 2.0 2.3

CV 78% 73% 56% 61% 62% 70% 123% 94%

Min. 0.32 0.33 0.50 1.1 0.32 0.33 0.17 0.81

Median 0.32 1.7 1.2 3.8 0.32 0.42 0.90 1.6

90th 0.79 3.7 2.5 6.5 0.56 1.1 3.9 4.8

Max. 1.5 4.4 2.8 8.3 1.2 1.6 7.8 10

Mean % of Total 12.3% 51.6% 36.8%

--- 16. 5% 22.0% 63.4%

Trial NT0 16 (n=20)* Trial NT0 17 (n=10)*

Olefin 5

- OH IMI Total Olefin 5

- OH IMI Total

Mean 0.60 0.40 1.5 2.5 0.32 0.33 0.96 1.6

SD 0.84 0.18 1.1 1.4 0.00 0.00 0.79 0.80

CV 141% 44% 75% 57% 0% 0% 83% 50%

Min. 0.32 0.33 0.50 1.1 0.32 0.3 3 0.17 0.81DP Barcode: D425260 MRID: 495 35601

10 Trial NT0 07 (n=15)* Trial NT008 (n=2 0) *

Olefin 5

- OH IMI Total Olefin 5

- OH IMI Total

Median 0.32 0.33 1.2 2.2 0.32 0.33 0.50 1.1

90th 1.1 0.50 3.0 4.5 0.32 0.33 1.6 2.2

Max. 4.0 1.1 3.7 6.0 0.32 0.33 2.8 3.4

Mean % of Total 23.7% 16.0% 60.8% 20% 21% 61%

N OTES : IMI= imidacloprid, olefin = imidacloprid olefin, 5

- OH = 5 hy droxy imidacloprid , Total = sum of IMI, olefin and 5

- OH (in bold). SD=standard deviation, CV = coefficient of variation.

* V alues below the LOD were assumed equal to 1⁄2 the LOD . Values below the LOQ (but above the LOD) were

assumed equal to 1⁄2 the LOQ.

Plot s of the individual measured residues of total IMI in nectar for each trial year are shown in Figure 2A

and 2B. Within each trial, it can be seen that applications during late summer/early fall resulted in

greater residues measured in nectar during year 1 (2013) compared to applications made in late spring / early summer as subsequent measurement in year 2 (2014). Mean values by residue trial and year are

shown in Table 7 below. Mean residues measured during year 1 from the NY sites (3.9

- 4.8 ppb) are about
4X higher than those measured during year 2 and that from the year 1 of the OR site (3.4 ppb for NT016)
is 2X the mean value measured during year 2 (1.7 ppb). The lower residue values measured during year 2
may reflect the greater amount of time available for dissipation of IMI residues between application and
residue measurement (275

- 300 days) compared to that from year 1 (about 205

- 215 days). Lower residues
in year 2 may also reflect temporal differences in imidacloprid translocation to nectar when appl ied in late summer/early fall vs. spring/early summer. It should be noted that except for 1 value from trial NT016,
residues of total IMI in nectar from year 2 (2014) largely reflect levels below the limit of analytical
detection or quantification where 1⁄2 the LOD or LOQ was assumed.

Table 7 . Mean total IMI residue in cherry nectar (ppb) by trial and study year.

Study Year Residue Trial

NY
( NT007 ) NY

( NT008 ) OR

( NT016 ) OR

( NT017 )

Year 1 (2013) 4.8 3.9 3.4 1.6

Year 2 (2014) 1.4 1. 1 1. 7 n.d.DP Barcode: D425260 MRID: 495 35601

11 Figure 2 . Total imidacloprid residues measured in cherry nectar for Trials NT007, NT008 (panel A) and
NT016, NT017 (panel B).

Pollen. Mean residues of total IMI in cherry pollen are 1

- 2 orders of magnitude grea ter than those

measured in nectar ( Table 7 ). Mean residues of total IMI pollen vary from 9.3

– 220 ppb while maxima
range from 3 8

- 1000 ppb

.

Table 8

. Magnitude of IMI and related metabolites in cherry pollen (ppb).

Trial NT007 (n=15)* Trial NT008 (n=20)*

Olefin 5

- OH IMI Total Olefin 5

- OH IMI Total

Mean 5.4 6.9 97 109 8.9 7.4 203 220

SD 3.6 3.6 84 91 9.9 7.0 259 273

CV 66% 53% 86% 83% 111% 95% 127% 124%

Min. 0.50 1.7 3.6 5.8 0.16 0.50 5.9 6.9

Median 5.1 7.3 85 99 4.4 4.4 84 96

90th 7.7 9.8 166 180 22 16 524 566

Max. 15 15 320 350 32 23 970 1000

Mean % of Total 5.0% 6.3% 88.6% 4.1% 3.4% 92.4%

Trial NT016 (n= 1 2)* Trial NT017 (n= 6 )*

Olefin 5

- OH IMI Total Olefin 5

- OH IMI Total

Mean 0.95 1.2 7.1 9.3 0.35 0.84 20 21

SD 1.9 2.0 9.9 11.1 0.47 0.48 24 24

CV 198% 168% 139% 119% 133% 57% 120% 116%

Min. 0.16 0.24 0.50 1.00 0.16 0.24 1.4 1.8

Median 0.16 0.24 4.6 5.0 0.16 0.85 11 12

90th 1.5 2.9 9.8 22.9 n.d. n.d. n.d. n.d.

Max. 6.7 7.0 37 38 1.3 1.3 65 67

Mean % of Tota l 10.2% 12.9% 75.9% 1.7% 4.0% 94.8%

N OTES : IMI= imidacloprid, olefin = imidacloprid olefin, 5

- OH = 5 hydroxy imidacloprid , Total = sum of IMI, olefin and 5

- OH (in bold). SD=standard deviation, CV = coefficient of variation. “n.d.” = not determined due t o small sample size
* Values below the LOD were assumed equal to 1⁄2 the LOD. Values below the LOQ (but above the LOD) were

assumed equal to 1⁄2 the LOQ.DP Barcode: D425260 MRID: 495 35601

12 It is clear from Table 9 and Figure s 3A and 3B that total IMI in pollen from the NY study sites are abou t
10X greater than those from the OR sites for both sampling years. Notably, such differences were not observed for nectar residues (Table 7). However, it is evident that the NY sites were not irrigated during
the trial period while sites in OR were irri gated. The influence of irrigation on IMI residues in pollen is not
known and other factors may account for the differences between the NY and OR results. Although the
basis of the 10X difference in pollen residue values between the NY and OR sites is no t clear, such

differences demonstrate the value of incorporating regional differences in study sites into the design of pollen/nectar residue studies.



### Table 9

. Mean total IMI residue in cherry pollen (ppb) by trial and study year

.

Study Year Residue Trial

NY
(NT007) NY

(NT008) OR

(NT016) OR

(NT017)

Year 1 (2013) 153 422 13.1 21

Year 2 (2014) 25 18 1.7 n.d.

Figure 3 . Total imidacloprid residues measured in cherry pollen for Trials NT007, NT008 (p anel A) and
NT016, NT017 (panel B).DP Barcode: D425260 MRID: 495 35601

13 Flowers

. Across all trials, m ean residues of total IMI in flowers range from 9.1

- 159 ppb in 2013 and 4.5

- 41
ppb in 2014 ( Table 10 ). Among all trials in 2013, mean residues in flowers were somewhat lower than
those in p ollen but the reverse is true for flower residues in 2014. Similar to the result for pollen, the NY
study sites contained residues in flowers that were about 10X those reported for the OR sites. As
described earlier, this same pattern of higher residues f rom the NY trial sites vs. OR sites was not evident

for nectar. Plots of the raw data for residues in flowers are provided in Figure s 4A and 4B

.

Table 10

. Mean concentrations of total IMI in cherry flowers (ppb).

Study Year Resid ue Trial

NY
(NT007) NY

(NT008) OR

(NT016) OR

(NT017)

Year 1 (2013) 98 159 12 9.1

Year 2 (2014) 38 41 4.5 n.d.

Figure 4

. Total imidacloprid residues measured in cherry flowers for Trials NT007, NT008 (panel A) and

NT016, N T017 (panel B).DP Barcode: D425260 MRID: 495 35601

14 6. C

. M AGNITUDE OF R ESIDUES IN L EAVES AND S OIL

Interestingly, mean residues of total IMI measured in leaves ( Table 1 1 ) are lower than those measured for
pollen and flowers (Tables 9 and 10, respectively), despite applications in 2012 per formed when foliage
was present (BBCH 91). This differs from results of other residue trials of foliar applications whereby
concentrations in leaves are highest among the matrices sampled (e.g., Citrus, MRID 49521301). R esidues

in leaves from the NY stud y sites were higher than those from the OR sites.

Table 11

. Magnitude of total IMI in c herry leaves (ppb).

Study Year Residue Trial

NY
(NT007) NY

(NT008) OR

(NT016) OR

(NT017)

Year 1 (2013) 64 53 11 9.3

Year 2 (2014) 34 38 5.0 n.d.

Unlike the other plant matrices sampled, the leaf residue data does span 6

- 17 days depending on the year
of sampling. As seen in Figure 5A , concentrations of total IMI in leaves from the NY sites decline
substantially over the two sampling events e ach year. Leaf residues measured from the OR sites do not
show an obvious decline, although concentrations are much lower overall ( Figure 5B ).

Figure 5

. Total imidacloprid residues measured in cherry leaves for Trials NT007 , NT008 (panel A) and
NT016, NT017 (panel B).DP Barcode: D425260 MRID: 495 35601

15 Residues of IMI measured in soil are low relative to most plant matrices, which is expected given the foliar
method of pesticide application (Table 1 2 )

. Levels were generally comparable across the four trials

. Carry
over from year to year was not evident based on sampling at the end of each growing season.

Table 12

. Magnitude of IMI residues in soil (ppb).

Year Date Event DAFA 1 Mean IMI in Soil

(ppb) 2

NT007

1 8/28/12 Prior to treatm ent

- 3 1 2

1 8/31/12

– 10/03/12 Foliar applications

1 5/10/13 End of 1 st growing season 2 52 113

2 6/5/13

– 7/15/13 Foliar applications

2 7/22/13 After last treatment 325 100

2 5/27/14 End of 2 nd growing season 634 62

NT008

1 8/31/12 Prior to trea tment

- 3 <1

1 9/3/12

– 10/8/12 Foliar applications

1 5/10/13 End of 1 st growing season 249 61

2 6/5/13

– 7/15/13 Foliar applications

2 7/22/13 After last treatment 322 57

2 5/27/14 End of 2 nd growing season 631 50

NT016

1 8/10/12 Prior to treatm ent

- 1 26

1 8/11/12

– 9/13/12 Foliar applications

1 5/2/13 End of 1 st growing season 264 53

2 6/5/13

– 7/11/13 Foliar applications

2 7/15/13 After last treatment 338 98

2 4/28/14 End of 2 nd growing season 625 57

NT017

1 8/10/12 Prior to treatmen t

- 1 <1

1 8/11/12

– 9/13/12 Foliar applications

1 4/29/13 End of 1 st growing season 261 21

2 5/28/13

– 7/1/13 Foliar applications

2 7/4/13 After last treatment 327 84

1 DAFA = Days After First Application in Year 1

2 Mean of 3 soil composite sampl es for IMI only

6. D

. Y EAR TO Y EAR A CCUMULATION IN P OLLEN AND N ECTAR

A qualitative evaluation of the potential for ‘year

- to

- year’ accumulation of total IMI residues in nectar and

pollen could be conducted for 3 of the 4 trials, since data from year 2 we re missing from trial NT017.

Examination of Figure s 2 and 3 (and Table 8 and 9) indicates that total IMI residues in nectar and pollen
decline from year 1 to year 2 . Similarly, residues of total IMI in flowers and leaves also decline from year

1 to year 2. Although this suggests that year

- to

- year accumulation of total IMI is not evident, the impact
of differences in pesticide application timing between year 1 to year 2 on the resulting residues in pollen

and nectar is not clear. In other words, the lowe r residues measured in year 2 may reflect longer timeDP Barcode: D425260 MRID: 495 35601

16 between pesticide application and residue sampling (and/or differences in pesticide translocation rates as
a function of application timing).

6. E

. R ESIDUE D ECLINE (DT 50 ) IN P OLLEN AND N ECTAR

Due t o the limited temporal coverage of residue sampling (2 sampling events/year with short sampling

intervals ), calculation of reliable dissipation half

- life (DT 50 ) values could not be conducted for nectar and
pollen.

6. F

. O CCURRENCE OF R ELEVANT IMI M ETA BOLITES

Nectar. Due to the low levels of parent IMI and metabolite residues measured in nectar , reliable

estimates of the relative proportion of total IMI as parent or metabolites could not be determined, since

such estimates would be unduly influenced by assumptions of 1⁄2 the LOD or 1⁄2 the LOQ.

Pollen. Suitable data for estimating t he proportion of total IMI residues in pollen present as parent IMI or

metabolites were available from Trials NT007 and NT008. In these trials, the mean % of total IMI as IM I

olefin in pollen is 6% (range 2

- 17%) . The mean % of total IMI as 5

- OH IMI is 8% (range 2

- 29%) . Parent IMI

was present at 86% of total IMI in pollen on average ( range 61

- 97% ).

6. F

. R ELATIONSHIP OF R ESIDUES A MONG P LANT T ISSUE S

Predictability of Flo wer Residues. The relationship between mean residues of total IMI measured in

flowers and total IMI measured in nectar and pollen on the same sampling day is shown in Figure 6 A and

6 B , respectively. Total IMI residues in nectar are predicted within 2X for 12 out of 13 paired values using

simple linear regression with flower data , although the r 2 (0.30) indicates a relatively weak correlation

( Panel A ). It is likely that assumptions regarding nectar concentrations measured below the LOD or LOQ

confounded t his relationship. T he relationship to total IMI residues in pollen vs. flowers is stronger than

that observed with the nectar data (r 2 =0. 6 4, Panel B )

. However, this correlation likely reflects the

occurrence of two large values recorded for NT008 and onl y 4/13 predicted resides in pollen are within 2X

of the observed values in pollen.

ADP Barcode: D425260 MRID: 495 35601

17 Figure 6

. Relationship between total IMI residues in flowers to those in nectar (A) and pollen (B)

Predictability of Leaf Residues

. The r elationship between mean residues of total IMI measured in cheery
leaves and total IMI measured in pollen and nectar on the same sampling day is shown in Figure 7

. A
correlation was not made between nectar and leave residues because of the very low values measured in
nectar and small sample size. Total IMI residues in pollen are predicted within 2X for only 2 of the 7
paired pollen and leave values using simple linear regression with leaf data, and the overall relationship is
relatively weak (r 2 = 0 .51 ).

Figure 7

. Relationship between total IMI residues in pollen to those in leaves

Pollen vs. Nectar

. The relationship between total IMI residues measured in c herry nectar and pollen is

shown in Figure 8 . A weak relationship is observed (r 2 =0. 44 ) and the range of nectar concentrations is
quite small compared to that in nectar. BDP Barcode: D425260 MRID: 495 35601

18 Caution is recommended when interpreting and applying these inter

- tissue regression relationships to
other situations since values here represent mean s of multiple composite samples and 2 of the 3 trials
had nearly identical weather and soil conditions.

Figure 8

. Relationship between total IMI residues in nectar to those in pollenDP Barcode: D425260 MRID: 495 35601

19 

### 7. STUDY STRENGTHS, LIMITATIONS AND CONCLU SIONS

In the context of documenting the magnitude of IMI residues in bee

- related matrices of c herry , the
following strengths are observed with this study.

1. Analytical methods were sufficiently accurate and precise based on QA data

2. Concentrations were meas ured for toxicologically

- relevant metabolites in multiple plant matrices

3. Application methods and rates were well documented

4. Samp le replication and compositing were reasonable to account for natural variability

5. Residues were measured over two growing season s to enable carry over to be assessed

. 6. Sites were located in two different parts of the U.S. (NY, OR) with widely differing climate

conditions

A number of limitations are also noted with this study, including:

1. Each of the two trials in NY and OR very clos e to each other thereby limiting the extent to which
differences in weather conditions and soil properties could be assessed within each region

. 2. The limited temporal s ampling did not enable calculation of DT 50 values

3. Differences in the timing of pesticid e applications between year 1 and year 2 may confound the
assessment of year to year carry over.

4. Some samples were not taken for certain trials (NT007, NT017) which limited their utility

Overall, considering the strengths and limitations of this study, the following conclusions can be drawn:

1. P ost

- bloom, foliar application of approximately 0. 1 lb a.i./A x 5 result in residues of total IMI in
individual composite samples of cherry nectar during the following year up to 10 ppb

. 2. P ost

- bloom, foliar applicat ion of approximately 0. 1 lb a.i./A x 5 result in residues of total IMI in
individual composite samples of cherry pollen during the following year up to 1 00 0 ppb

. 3. Mean concentrations of total IMI in cherry nectar measured after foliar applications during t he

previous late summer/early fall (Year 1) were about 2X those measured after foliar applications

during late spring/early summer (year 2) across all trials. A larger difference was observed in

mean pollen concentrations (about 10X) across trial years

. It is not clear if this finding reflects
seasonal differences in IMI translocation or longer intervals between pesticide application and
residue measurement associated with year 2.

4. Mean t otal IMI residues in pollen were approximately 1

- 2 orders of magnitud e greater than those

observed in nectar

. 5. Mean concentrations of total IMI measured in pollen from the NY study trails were about 10X
those measured in pollen from the OR trial sites. Similar results were observed with total IMI

residues measured in flower s and leaves.

6. Two metabolites of IMI (IMI

- olefin and 5

- OH

- IMI) typically represented < 1 0 % of total IMI residues
in pollen and nectar.

7. The predictability of t otal IMI residues in nectar and pollen based on measured concentrations in
other plant matrices ( f lowers , leaves) was generally poor.



### 8. STUDY VALIDITY/CLASSIFICATION

Data from the four trials (NT00 7, NT008, NT016 and NT017) are considered scientifically sound and useful
for risk assessment purposes, although these trials are highly similar in m any respects do to their close

proximity within the NY and OR study regions

. Overall, this study is classified as ACCEPTABLE for
quantitative use in risk assess ment.DP Barcode: D425260 MRID: 495 35601

20 Appendix A. Location of C herry Residue Trials.

Figure A1. Locations of Trial NT00 7 (A lton, NY) and NT008 ( Lyons, NY )

Figure A 2

. Locations of Trial NT016 and NT017 (Mosier, OR) 

### NT008 NT007