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Crop Protection
journal homepage: www.elsevier.com/locate/cropro
A R T I C L E
I N F O
A B S T R A C
T
Article history:
Received 27 May 2011
Received in revised form
29 November 2011
Accepted 11 December
2011
Keywords:
Pesticides
Droplet size
Variability factor
Degradation
Composite samples
Fruit
In this study, field trials on fine and coarse spray quality application of pesticides on apples were
performed. The main objectives were to study the variation of pesticide residue levels in individual
fruits
versus composite samples, and the effect of standard ine spray quality application versus coarse
spray
quality application on residue levels. The applications included boscalid, bupirimate, captan,
fenoxycarb,
indoxacarb, pirimicarb, pyraclostrobin and thiophanate-methyl. Apples were collected from four
zones
in
the tree and pesticide residues were detected in the individual apples. None of the results for the
pesticides
residues measured in individual apples exceeded the EU Maximum Residue Levels (MRLs). However,
there
was a large variation in the residues levels in the apples, with levels from 0.01 to 1.4 mg kg -1 for
captan,
the
pesticide with the highest variation, and from 0.01 to 0.2 mg kg -1 for pyraclostrobin, the pesticide
with
the
lowest variation. Residues of fenoxycarb and indoxacarb were only found in a few apples, probably
due
to
the early application time of these two compounds. The evaluation of the effect of spray quality did
not
show any major difference between fine and coarse spray quality, except for carbendazim, the
degradation
product of thiophanate-methyl, where ine spray quality resulted in higher carbendazim residue
levels
than coarse spray quality. To examine the relationship between individual results and average
results
from
ten apples, 20 composite samples were statistically constructed from sets of ten of the individual
results.
The variability factors for the individual samples (n = 80) at the 97.5 percentile were calculated for
both
standard and air induction nozzle application and were in the range of 0.99.4. The variability factor
of
seven used when EU member states calculate possible exceeding of Acute Reference Dose (ARfD)
was
adequate to encompass almost all the average results from the analyses of ten individual apples.
However,
for captan up to 9% of the results were not covered depending on which of the mathematically
constructed
composite concentrations was chosen. The variability factor of three, recommend by Codex, seems
to
be
too low, because up to 30% of the apple samples for captan were not covered if the worst case
scenario
was
chosen. The factor of three seems was also too low for thiophanate-methyl.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
* Corresponding author. Tel.: +45 3588 7463; fax: +45 3588 7448.
E-mail address: mpou@food.dtu.dk (M.E. Poulsen).
0261-2194/$ see front matter 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.cropro.2011.12.013
shown
to
affect variability in initial deposit. Canopy structure is one of
the
most important factors influencing pesticide residues. Many
studies
2010).
In
practice coarse droplet applications might result in more
visible
residue spots on apples and pears (fruits). However, it is
unknown
if
this also affects the residue levels of plant protection products
(PPPs)
in fruits as well. Because of the high values of spray drift in
orchard
spraying (Zande et al., 2001) compared to arable ield
applications
(Huijsmans et al., 1997), the reduction of the emission of PPPs
in
fruit
growing is still of major importance. One of the measures is
the
use
of
drift reducing nozzles (Zande et al., 2008). It is a relatively
simple
and
cheap method. The use of drift reducing nozzles in fruit
growing
has been increased in many countries in Europe, e.g. Belgium,
France,
Germany and the Netherlands. Drift reducing nozzles are
often
accepted within legislation for fruit growing; e.g. Germany
(JKI,
2010),
United Kingdom (HSE, 2010), and the Netherlands (CTGB,
2008).
A ield treatment with pesticides is not uniform and consequently the distribution of pesticide residues may be very
wide,
from
areas receiving high amounts of pesticides to areas receiving
less
amount of pesticide. In 1994 research carried out in the UK,
indicated
that there might be occasional and random high residues of
pesticides found in individual samples of fresh fruit and vegetables
(Harris et al., 2000; Harris, 2000). The finding of high residue
variability is considered important in the assessment of acute
dietary
exposure, as it might lead to a higher intake of pesticide
residues
than previously believed. Field trial data derived from
commercial
use of pesticides indicates that the distribution of residues is
not
signiicantly inluenced by the 1) mean value of the residue, 2)
time
between pesticide application and sampling, 3) chemical and
physical properties of the active ingredient or formulation or
4)
the
volume application rate used for foliar application on fruit
trees.
On the other hand the distribution of residues is likely to be
inluenced by the size, shape and density of the plants and mode
of
application of the pesticide (Ambrus, 2000). Research on the
causes
of residue variability has been conducted by Carter et al.
(2000),
but
no conclusive evidence could be obtained which identify
particular
causes for the variability. Hill and Reynolds (2002)
demonstrated
the row). Both plots were sprayed for the whole season with
the
prescribed nozzle type, according to a standard commercial
spraying
scheme with insecticides and fungicides (Table 1), following
label
directions. Dose rates were therefore kept equal for the Albuz
ATR
and Albuz TVI nozzles.
The distribution in different zones of the canopy was
evaluated
by dividing the apple tree (spindle type) into four distinct
zones:
top, middle, bottom outside and bottom inside (Fig. 1). Four
apples
were collected from five trees per nozzle type and tree zone.
Selection of the trees and apples per zone was randomized
from
each tree row (variety Elstar) in the middle of the
experimental
plot. In summary, the levels in the ield sampling used were:
(1)
fruits within a zone, (2) zone within a tree, (3) trees within a
nozzle
type. Two days before the start of commercial harvest, single
fruit
samples were taken from ive trees for each zone and nozzle
type.
The total weight of each individual apple was determined
before
the residue analysis.
2.2. Chemicals
The pesticide standards of boscalid, bupirimate, captan,
Fig. 1. Four zones within the apples trees from where apples were sampled. T: Top,
M: Middle, BU: Bottom Outside, BI: Bottom Inside.
Table 1
Application date, pesticide formulations and active ingredients.
Application date
8 May 2007
14 May 2007
19 May 2007
25 May 2007
2 June 2007
6 June 2007
11 June 2007
Pesticide product
Merpan
Merpan
Merpan
Merpan
Merpan
Merpan
Merpan
Nimrod
Active ingredients
Captan
Captan
Captan
Captan
Captan
Captan
Captan
Bupirimate
Insegar
Fenoxycarb
Merpan
Nimrod
Captan
Bupirimate
Insegar
Fenoxycarb
19 June 2007
Merpan
Steward
Captan
Indoxacarb
25 June 2007
30 June 2007
8 July 2007
15 July 2007
19 July 2007
27 July 2007
04 August 2007
13 August 2007
Merpan
Merpan
Merpan
Merpan
Bellis
Merpan
Merpan
Bellis
Merpan
Captan
Captan
Captan
Captan
Boscalid + Pyraclostrobin
Captan
Captan
Boscalid + Pyraclostrobin
Captan
19 August 2007
Merpan
Pirimor
Captan
Pirimicarb
25 August 2007
Merpan
Topsin-M
Captan
Thiophanate-methyl
16 June 2007
* : Pirimor was sprayed to control aphids in the lower parts of the trees. The top
nozzles of the sprayer were closed in order to avoid unnecessary residues.
11
Table 2
MRM transitions, cone voltages, collision energies and recoveries for LC/MS/MS and GC/MS/MS analysis of boscalid, bupirimate, carbaryl 13-C6, carbendazim, captan,
chlorpyrifos D10, fenoxycarb, indoxacarb, pirimicarb, pyraclostrobin, thiophanate-methyl and tetrahydrophthalimide. Carbaryl 13-C6 and chlorpyrifos D10 were used as
injection standards and consequently no recoveries are listed for these compounds.
GC/MS/MS
LC/MS/MS
Instrument Compound
Transistion1 Cone Voltage Collision energy Transistion2 Cone Voltage2 Collision Energy2 Recovery at 0.02 mg kg
Boscalid
343 >307 57
20
343 > 140 57
20
93%
Bupirimat
317
Carbaryl 13-C6
225
>166
>151
27
23
317
25
14
208
> 108
> 151
25
50
50
14
Carbendazim
192
Fenoxycarb
>160
>88
80
15
192
> 132
80
15
302
25
20
>
Indoxacarb
529
>218
65
20
>
Pirimicarb
239
>72
25
16
239
Pyraclostrobin
388
24
11
388
Thiophanate-methyl
343
50
12
343
Bupirimate
316
>194
>151
>208
15
273
> 163
> 93
> 193
Captan
149
>70
12
149
Chlorpyrifos D10
326
>262
15
292
Pirimicarb
238
>166
10
Tetrahydrophthalimide
151
>80
76%
85%
95%
> 182
25
14
24
25
98%
50
12
101%
10
92%
> 105
90%
> 260
10
166
> 96
15
87%
123
> 80
97%
94%
90%
in individual apples
Residues of bupirimate were detected in almost all the
apple
samples but less than 15% of the residues were above the
Limit
of
Quantiication (LOQ) of 0.005 mg kg -1. Pirimicarb was not
sprayed
by
the top nozzles (see Table 1), because only the lower parts of
the
trees
were infested by aphids, and consequently pirimicarb was not
found
in 36 apples picked from the top of the trees. Residues of
fenoxycarb
and indoxacarb were only found in few apples probably due to
the
early application time for those two compounds. Statistical
evaluation
of bupirimate, pirimicarb, fenoxycarb and indoxacarb is
therefore
not
included in this presentation. For carbendazim, residues were
detected above the LOQ (0.01 mg kg-1) in 156 of the 160 apples. All
four
apples with no detectable residues were sprayed by the air
induction
nozzle; two were picked from the top and two from the
bottom
inside.
Thiophanate-methyl residues were not detected above the
LOQ
(0.01 mg kg-1) in three apples, i.e. one air induction nozzle
application
top and two standard application bottom outside. Boscalid,
pyraclostrobin, captan and the degradation product
tetrahydrophthalimide were found in all 160 samples (LOQ = 0.01 mg kg -1).
This
is
in accordance with Xu et al. (2008) who found that initial
captan
deposits and subsequent residues on fruits and leaves are
generally
closer to a lognormal than to a normal distribution.
None of the results for the pesticides residues measured in
the
apples exceeded the MRLs (European Commission, 2010).
However,
12
Table 2
higher
mean values at the middle zone, while tetrahydrophthalimide
had
higher levels for the bottom inside zone. For carbendazim the
standard application resulted in higher mean values at all
zones.
More details are described below under the active
substances.
3.1.3.
Zone effect
The mean values for the four zones have been statistically
evaluated to see if one of the application modes gave a more
even
distribution (Fig. 3). The evaluation for boscalid and
pyraclostrobin
showed that the pesticides seem to be more evenly
distributed
by
the
air induction nozzle application, while it was opposite for
carbendazim. However, no major differences were seen. Xu et al.
(2008)
showed that the unit-to-unit variation contributes most to the
observed variability in captan residues, not only in the initial
spray
deposition but also over the subsequent period following the
application. More details are described below under the active
substances.
3.1.4.
Boscalid and pyraclostrobin
Boscalid and pyraclostrobin were sprayed simultaneously
(as they were active ingredients of the fungicide product
Bellis)
13
0.00
b
Boscalid
mg/kg
Whole tree BI
BU
Zone
Whole tree
0.10
mg/kg
Captan
BI
0.25
0.20
BU
Zone
0.08
BU
Zone
Thiophanate-methyl
0.15
BI BU
Zone
Whole tree BI
0.06
0.04
0.02
Pyraclostrobin
0.00
ME. Poulsen et al. /
Crop
Protection
35
(2012)
5-14
standard
mg/kg
air induction nozzle
0.10
0.35
0.10
Whole tree
Zone
BI
BU
0.08
0.30
0.08
0.25
0.06
0.00
0.06
0.20
0.04
0.15
0.04
mg/kg
0.10
0.02
0.02
0.05
0.00
0.00
0.35
'
0.05
0.30
0.25
0.20
0.15
0.10
0.05
Carbendazim
Tetrahydrophthalimide
mg/kg
0.10
b
b
14
Whole tree
BI
BU
M
T
15
Zone
Fig. 2. Effect of nozzle types (spray quality); Mean values of pesticide residues of boscalid, pyraclostrobin, captan and its degradation compound tetrahydrophthalimide,
carbendazim and thiophanate-methyl from analysis of individual apples picked at four zones within the tree. BI: bottom inside, BU: bottom outside, M: middle, T: top. The
mean
values for all apples are also shown. White bars are standard application Albuz ATR nozzle (fine spray quality); black bars are air induction nozzle application (coarse spray
quality).
Bars labeled with the same letter do not differ significantly p = 0.05.
Northover
et al., 1986). The monograph on captan by FAO/WHO (1970)
also
reports that captan does not appear to undergo any chemical
change on the plant surface. However, Ticha et al. (2007)
showed
captan residues in apples successively decreased until
harvest.
None of the above quoted studies had measured the
degradation
product tetrahydrophthalimide. In this study tetrahydrophthalimide was found in higher levels than captan. The results
showed that approximately 80% of the sum of captan and
tetrahydrophthalimide was found as tetrahydrophthalimide, when
taking the differences in molecular weights into account. The
results are in line with results from Poulsen et al. (2009),
where
7
different spraying strategies were performed in commercial
orchards. However, part of the degradation could occur in the
laboratory during the chemical analysis. Captan is rapidly
degraded
16
Pyraclostrobin
0.00
0.15
0.20
0.25
0.10
Boscalid
BI BU
standard
BI BU
M
air induction
mg/kg
T BI BU M
air induction
17
Standard
mg/kg
0.35
0.30
Captan
0.2
b
5
0.2
0
0.1
5
0.1
- 1
F
Iii
IX
M
BI
BU
air induction
BI
BU
BI BU
standard
T
BI BU M
air induction
18
BU
mg/kg
M
0.10
standard
Th ioph an ate-methyl
0.08
---------c
0.06
0.04
0.02
BU M
standard
BI BU M
air induction
Carbendazim
bb
Al
0.00 Bl
I
Bl
BU
air induction
19
Fig. 3. Effect of the zone within the tree; mean values of pesticide residues of boscalid, pyraclostrobin, captan and its degradation compound tetrahydrophthalimide,
thiophanatemethyl and carbendazim, from analysis of individual apples picked at four zones. BI: bottom inside, BU: bottom outside, M: middle, T: top. Results for standard application
(fine spray quality) and air induction nozzle application (coarse spray quality) are shown bars labeled with the same letter do not differ significantly p = 0.05.
-1
20
Table
3
mg/kg
Acceptable Daily Intake (ADI) and Acute Reference Dose (ARfD) in mg kg -1 body
weight for boscalid, pyraclostrobin, captan, tetrahydrophthalimide, carbendazim
and thiophanate-methyl.
Acceptable Daily Intake,
0.04
none
Bupirimate
0.05
0.05
Captan
0.1
Carbendazim
0.02
0.02
Fenoxycarb
0.04
none
Indoxacarb
0.006
0.02
Pirimicarb
0.035
Pyraclostrobin
0.03
Tetrahyd
rophthal
imide
0.3
0.1
0.03
Tetrahydrophthalimide
none found
Thiophanate-methyl
0.08
none found
0.2
Fine sprayquality
Coarse sprayquality
X
Boscali
d nX
mg/k
0.60g
0.30
0.50
0.20
0.40
0.10
0.00
Captan
Composite
Individual
Pyraclostrobin
X
mg/k
0.25g
Composit
Individua
0.20
0.15
0.10
0.05
i
I
Composite
Individual
Composite
-----------------------------------------1----------------------------------------------------------------------------,-------------------------------------1
mg/k
1.5 g
0
Fine sprayquality
0.80
0.60
Fine sprayquality
v
1.2
0.7
5
0.5
0
x
1.0
0
I
i
0.25
0.00
Composite
Composite
Individual
1.00
Composite
Individual
Composite
22
Table
3
Thiophanate-methyl
mg/k
0.30g
xx
0.25
0.15
0.10
0.05
Composite
K
Individual
0.20
0.15
1
Composite
Individua
l
0.25
0.20
0.00
Carbendazim
mg/k
0.30g
0.10
i
i
'
0.05
0.00
Composite
Individual
Composite
Individual
Fig. 4. Results for composite and individual samples for pesticide residues of boscalid, pyraclostrobin, captan and its degradation compound tetrahydrophthalimide,
carbendazim
and thiophanate-methyl. Results are shown for both standard application Albuz ATR nozzle (fine spray quality) and air induction nozzle application (coarse spray quality).
23
Table 4
Variability factor for standard and air induction nozzle application for boscalid,
captan,
carbendazim, tetrahydrophthalimide, thiophanate-methyl and pyraclostrobin.
Standard
application
Boscalid
Captan
Carbendazim
Tetrahydrophthalimide
Thiophanate-methyl
Pyraclostrobin
2.0
9.4
1.8
4.0
3.9
2.2
Air induction
nozzle application
2.7
4.3
0.9
2.7
3.8
2.6
The EU MRLs and CODEX MRLs for apples are set for the
mean
residue value of one kg or a minimum of ten apples (European
Commission, 2002; CODEX, 1999). Originally, the toxicological
evaluation of pesticides was performed for chronic effects
only
by
setting an Acceptable Daily Intake (ADI) (Table 3). For control
purposes of ADI an average residue value are more suitable in
order
to smooth out the variation in residue levels, because a
person
eating a high pesticide residue content apple one day will
most
likely eat an apple with at lower content the next day. Consequently, the control for pesticide residues in apples was
decided
to
be carried out for composite samples. Later on, the pesticides
were
evaluated for acute effects and Acute Reference Doses (ARfD)
were
set. To control the ARfDs it is important to know the pesticide
residue in the individual apple, because a person will be
exposed
to
acute effects from a speciic apple, if the residues are high
enough.
However, it would be costly to control the ARfD, if the apple
samples should be analyzed individually as ten apples should
be
analyzed. Instead, the pesticide residue levels in the
composite
samples are calculated back to the assumed highest possible
level
in the individual apples, by applying a variability factor to the
average concentration of ten apples. In EU member states the
variability factor used is seven (EU Commission, 2004),
whereas
CODEX has lowered the factor to a default value of three until
suficient data on single unit is available to calculate a more
realistic
variability factor (WHO, 2010).
To investigate the distribution of the pesticides between
the
apples from the two application modes, all apples were
individually
analyzed. The results of from the analysis of the individual
apples
showed large variations in residue levels, where the apples
with
the
highest concentration had between 12 and 200 times higher
residue
levels than in the apple with the lowest level. To examine the
rela-
Table 5
The minimum and maximum number of individual results not covered by the
variability
factor
3
and
7,
for
boscalid,
pyraclostrobin,
captan,
tetrahydrophthalimid,
carbendazim and thiophanate-methyl, obtained by standard and air induction
nozzle application. The number before the arrows refers to the highest concentration of the 20 mathematically constructed composite samples and the number
after
the arrow refers to the lowest concentration of 20 the mathematically constructed
composite samples. The mathematically constructed composite samples represent
a sample of ten apples used for control of MRLs.
Standard application
Boscalid
Pyraclostrobin
Captan
Tetrahydrophthalimid
Thiophanate-methyl
Carbendazim
Factor 3
0/1
0/1
7 / 24
0/8
2/8
0/0
Factor 7
0/0
0/0
1/4
0/0
0/0
0/0
Air induction
application
Factor 3
1/4
0/0
0/7
0/4
1/6
0/0
Factor 7
0/0
0/0
0/0
0/0
0/1
0/0
24
Table 4
composite
samples was used and one result (the highest) if calculated
with
the
highest result for the composite samples.
As seen from Table 5 the variability factor of seven seems
to
be
adequate to include almost all the results for the pesticides.
For
captan up to 9% of the results were not covered depending on
the
composite concentration chosen. However, the variability
factor
of
three seems to be too low, because up to 30% of the apple
samples
for captan were not covered if the worst case scenario was
chosen.
Also for thiophanate-methyl the factor of three seems to be
too low.
In conclusion, no statistically significant correlation
between
the
fruit weight (size) and residue level was observed, showing
that
the residue levels were independent of the fruit size. None of
the
results for the pesticides residues measured in the apples
exceeded
the EU MRLs. For boscalid and pyraclostrobin no differences
were
seen in the residue levels for the two spraying techniques.
The
coarse spray quality (air induction nozzle) resulted in higher
levels
for captan and thiophanate-methyl at the middle zone and for
tetrahydrophthalimide at the bottom inside zone. However,
with
25
26
Zande van de, J.C., Heijne, B., Wenneker, M., 2001. Spray Drift Reduction in
Orchard
Spraying (State-of-the-Art December 2001). Institute of Agricultural and Environmental Engineering/PPO-fruit. IMAG Report 2001-19, Wageningen. 38 p. (in
Dutch with English summary).
Zande van de, J.C., Holterman, H.J., Wenneker, M., May, 2008. Nozzle classification
for drift reduction in orchard spraying: identification of drift reduction class
threshold nozzles. In: Agricultural Engineering International: The CIGR Ejournal
Manuscript ALNARP 08 0014, vol. X.
28
Crop Protection
journal homepage: www.elsevier.com/locate/cropro
1 Corresponding author. Tel.: +45 3588 7463; fax: +45 3588 7448.
E-mail address: mpou@food.dtu.dk (M.E. Poulsen).
0261-2194/$ see front matter 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.cropro.2011.12.013
30
A R T I C L E
I N F O
A B S T R A C
T
Article history:
Received 27 May 2011
Received in revised form
29 November 2011
Accepted 11 December
2011
Keywords:
Pesticides
Droplet size
Variability factor
Degradation
Composite samples
Fruit
In this study, field trials on fine and coarse spray quality application of pesticides on apples were
performed. The main objectives were to study the variation of pesticide residue levels in individual
fruits
versus composite samples, and the effect of standard ine spray quality application versus coarse
spray
quality application on residue levels. The applications included boscalid, bupirimate, captan,
fenoxycarb,
indoxacarb, pirimicarb, pyraclostrobin and thiophanate-methyl. Apples were collected from four
zones
in
the tree and pesticide residues were detected in the individual apples. None of the results for the
pesticides
residues measured in individual apples exceeded the EU Maximum Residue Levels (MRLs). However,
there
was a large variation in the residues levels in the apples, with levels from 0.01 to 1.4 mg kg -1 for
captan,
the
pesticide with the highest variation, and from 0.01 to 0.2 mg kg -1 for pyraclostrobin, the pesticide
with
the
lowest variation. Residues of fenoxycarb and indoxacarb were only found in a few apples, probably
due
to
the early application time of these two compounds. The evaluation of the effect of spray quality did
not
show any major difference between fine and coarse spray quality, except for carbendazim, the
degradation
product of thiophanate-methyl, where ine spray quality resulted in higher carbendazim residue
levels
than coarse spray quality. To examine the relationship between individual results and average
results
from
ten apples, 20 composite samples were statistically constructed from sets of ten of the individual
results.
The variability factors for the individual samples (n = 80) at the 97.5 percentile were calculated for
both
standard and air induction nozzle application and were in the range of 0.99.4. The variability factor
of
seven used when EU member states calculate possible exceeding of Acute Reference Dose (ARfD)
was
adequate to encompass almost all the average results from the analyses of ten individual apples.
However,
for captan up to 9% of the results were not covered depending on which of the mathematically
constructed
composite concentrations was chosen. The variability factor of three, recommend by Codex, seems
to
be
too low, because up to 30% of the apple samples for captan were not covered if the worst case
scenario
was
chosen. The factor of three seems was also too low for thiophanate-methyl.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
Pesticides are widely used in the production of fruit. In
apple
production fungicides and insecticides are used pre harvest to
protect the apples from a range of pests and diseases and to
provide
quality preservation. Due to the potential risk of pesticide
residues
to human health the use of pesticides is strictly regulated,
e.g.
by
the
establishment of Maximum Residue Levels (MRLs). Many
countries
have monitoring programmes for pesticide residues in food to
ensure that pesticide residues do not exceed their MRLs.
Monitoring
programmes have shown that fungicide and insecticide
residues
are
often found in apples. In the EC coordinated monitoring
programme
32
factor
and the 1) commodity, 2) country of origin, 3) residue
concentration
or 4) physiochemical characteristics of the pesticide including
the
systemic/non-systemic
properties
of
the
pesticide.
Furthermore,
it was seen that aging of the residue did not signiicantly affect
the
variability. Consequently, variability may only be reduced in
terms
of
more uniform application of the pesticide. When assessing the
variability it is relevant to look at the precision of the
analytical
method, as variation might contribute to the variability of the
single
commodities. Investigations conducted by Harris and Davis
(1998)
and Hill and Reynolds (2002) showed that the analytical
variation
did not signiicantly contribute to the overall variation. Thus
the
main contributor to the variation must be the difference in
residue
content between the individual units.
Explanation of the variability in pesticide residues is
essential
for the development of methods to minimize residues in fruits.
In
this study, residue variability associated with individual apples
and
composite samples was studied for two application
techniques;
i.e.
ine and coarse spray quality application. It was hypothesized
that
variation among individual fruits is the most important factor
of
residue variation.
Field trials were performed in a commercial orchard in
2007
with application of pesticides routinely used for pest and
disease
management in apple production. Apples were collected from
4
zones in the tree.
The aims of this study were: (1) to determine residue levels
of
commonly applied pesticides in apple growing after ield treatment, (2) to study the variation of these residue levels in
individual
fruits and/versus (calculated) composite samples, and (3) the
effect of a standard ine spray quality versus coarse spray
quality
application on residue levels.
2. Materials and methods
2.1. Field work
A ield trial to evaluate the effect of spray quality (droplet
size/
nozzle type) was done in 2007 in a commercial orchard in the
Netherlands. The spray applications for the reference
treatment
(fine
spray quality) were performed with brown Albuz ATR hollow
cone
nozzles (7 bar spray pressure) and for the low-drift treatment
(coarse
spray quality) with air induction (Venturi) hollow cone nozzles
(Albuz TVI 80-015 at 6 bar spray pressure). Spray applications
were carried out in the same orchard, and with the same
spraying
machine; a Munckhof cross-low sprayer with eight operating
nozzles at each side. The power take-off (PTO) speed was ixed
at
430 rpm, and the driving speed was 8 km h -1. These settings
resulted
in a spray volume rate of approximately 200 l ha-1 for the
brown
Albuz ATR nozzles and 300 l ha -1 for the air induction nozzles.
The
standard advice to growers is to use a higher water volume
for
drift
reducing air induction nozzles (Jaeken et al., 2003). The spray
quality
of the nozzles was quantiied using a Phase Doppler Particle
Analyser
(PDPA, Aerometrics). These measurements showed that the
brown
Albuz ATR nozzle had a Volume Median Diameter (VMD) of
143
mm
and the Albuz TVI 80-015 had a VMD of 493 mm (Zande et
al., 2008).
The orchard was divided in two experimental plots. Each
plot
consisted of 8 apple tree rows; i.e. 4 times 2 alternating rows
of
the
apple varieties Elstar and Jonagold of 150 m length (with an
average
tree height of 3.00 m, 3 m row spacing, and 1.25 m tree
separation in
35
the row). Both plots were sprayed for the whole season with
the
prescribed nozzle type, according to a standard commercial
spraying
scheme with insecticides and fungicides (Table 1), following
label
directions. Dose rates were therefore kept equal for the Albuz
ATR
and Albuz TVI nozzles.
The distribution in different zones of the canopy was
evaluated
by dividing the apple tree (spindle type) into four distinct
zones:
top, middle, bottom outside and bottom inside (Fig. 1). Four
apples
were collected from five trees per nozzle type and tree zone.
Selection of the trees and apples per zone was randomized
from
each tree row (variety Elstar) in the middle of the
experimental
plot. In summary, the levels in the field sampling used were:
(1)
fruits within a zone, (2) zone within a tree, (3) trees within a
nozzle
type. Two days before the start of commercial harvest, single
fruit
samples were taken from ive trees for each zone and nozzle
type.
The total weight of each individual apple was determined
before
the residue analysis.
2.2. Chemicals
The pesticide standards of boscalid, bupirimate, captan,
Fig. 1. Four zones within the apples trees from where apples were sampled. T: Top,
M: Middle, BU: Bottom Outside, BI: Bottom Inside.
36
Table 1
Application date, pesticide formulations and active ingredients.
Application date
8 May 2007
14 May 2007
19 May 2007
25 May 2007
2 June 2007
6 June 2007
11 June 2007
Pesticide product
Merpan
Merpan
Merpan
Merpan
Merpan
Merpan
Merpan
Nimrod
Active ingredients
Captan
Captan
Captan
Captan
Captan
Captan
Captan
Bupirimate
Insegar
Fenoxycarb
Merpan
Nimrod
Captan
Bupirimate
Insegar
Fenoxycarb
19 June 2007
Merpan
Steward
Captan
Indoxacarb
25 June 2007
30 June 2007
8 July 2007
15 July 2007
19 July 2007
27 July 2007
04 August 2007
13 August 2007
Merpan
Merpan
Merpan
Merpan
Bellis
Merpan
Merpan
Bellis
Merpan
Captan
Captan
Captan
Captan
Boscalid + Pyraclostrobin
Captan
Captan
Boscalid + Pyraclostrobin
Captan
19 August 2007
Merpan
Pirimor
Captan
Pirimicarb
25 August 2007
Merpan
Topsin-M
Captan
Thiophanate-methyl
16 June 2007
*: Pirimor was sprayed to control aphids in the lower parts of the trees. The top
nozzles of the sprayer were closed in order to avoid unnecessary residues.
38
Table 2
MRM transitions, cone voltages, collision energies and recoveries for LC/MS/MS and GC/MS/MS analysis of boscalid, bupirimate, carbaryl 13-C6, carbendazim, captan,
chlorpyrifos D10, fenoxycarb, indoxacarb, pirimicarb, pyraclostrobin, thiophanate-methyl and tetrahydrophthalimide. Carbaryl 13-C6 and chlorpyrifos D10 were used as
injection standards and consequently no recoveries are listed for these compounds.
GC/MS/MS
LC/MS/MS
Instrument Compound
Transistionl Cone Voltage Collision energy Transistion2 Cone Voltage2 Collision Energy2 Recovery at 0.02 mg kg
Boscalid
343 >307 57
20
343 > 140 57
20
93%
Bupirimat
317
Carbaryl 13-C6
225
>166
>151
27
23
317
25
14
208
> 108
> 151
25
50
50
14
Carbendazim
192
Fenoxycarb
>160
>88
80
15
192
> 132
80
15
302
25
20
>
Indoxacarb
529
>218
65
20
>
Pirimicarb
239
>72
25
16
239
Pyraclostrobin
388
24
11
388
Thiophanate-methyl
343
50
12
343
Bupirimate
316
>194
>151
>208
15
273
> 163
> 93
> 193
Captan
149
>70
12
149
Chlorpyrifos D10
326
>262
15
292
Pirimicarb
238
>166
10
Tetrahydrophthalimide
151
>80
76%
85%
95%
> 182
25
14
24
25
98%
50
12
101%
10
92%
> 105
90%
> 260
10
166
> 96
15
87%
123
> 80
97%
94%
90%
in individual apples
Residues of bupirimate were detected in almost all the
apple
samples but less than 15% of the residues were above the
Limit
of
Quantification (LOQ) of 0.005 mg kg -1. Pirimicarb was not
sprayed
by
the top nozzles (see Table 1), because only the lower parts of
the
trees
were infested by aphids, and consequently pirimicarb was not
found
in 36 apples picked from the top of the trees. Residues of
fenoxycarb
and indoxacarb were only found in few apples probably due to
the
early application time for those two compounds. Statistical
evaluation
of bupirimate, pirimicarb, fenoxycarb and indoxacarb is
therefore
not
included in this presentation. For carbendazim, residues were
detected above the LOQ (0.01 mg kg-1) in 156 of the 160 apples. All
four
apples with no detectable residues were sprayed by the air
induction
nozzle; two were picked from the top and two from the
bottom
inside.
Thiophanate-methyl residues were not detected above the
LOQ
(0.01 mg kg-1) in three apples, i.e. one air induction nozzle
application
top and two standard application bottom outside. Boscalid,
pyraclostrobin, captan and the degradation product
tetrahydrophthalimide were found in all 160 samples (LOQ = 0.01 mg kg -1).
This
is
in accordance with Xu et al. (2008) who found that initial
captan
deposits and subsequent residues on fruits and leaves are
generally
closer to a lognormal than to a normal distribution.
None of the results for the pesticides residues measured in
the
apples exceeded the MRLs (European Commission, 2010).
However,
39
Table 2
higher
mean values at the middle zone, while tetrahydrophthalimide
had
higher levels for the bottom inside zone. For carbendazim the
standard application resulted in higher mean values at all
zones.
More details are described below under the active
substances.
3.1.3.Zone effect
The mean values for the four zones have been statistically
evaluated to see if one of the application modes gave a more
even
distribution (Fig. 3). The evaluation for boscalid and
pyraclostrobin
showed
that
the
pesticides
seem
to
be
more
evenlydistributed
bythe
air induction nozzle application, while it was opposite for
carbendazim. However, no major differences were seen. Xu et al.
(2008)
showed that the unit-to-unit variation contributes most to the
observed variability in captan residues, not only in the initial
spray
deposition but also over the subsequent period following the
application. More details are described below under the active
substances.
3.1.4.Boscalid and pyraclostrobin
Boscalid and pyraclostrobin were sprayed simultaneously
(as they were active ingredients of the fungicide product
Bellis)
40
Boscalid
Pyraclostrobin
M
.
E
.
0.00
P
o
u
l
s
mg/kg
Whole tree BI
BU
Zone
Whole tree
0.10
mg/kg
Captan
BI BU
Zone
T
n
e
t
0.08
BI
0.25
0.20
BU
Zone
Thiophanate-methyl
0.15
Whole tree BI
0.02
0.00
0.05
Whole tree
Zone
0.00
mg/kg
BI
BU
T
.
/
0.06
0.04
0.10
BU
Zone
C
r
o
p
P
r
o
t
e
c
t
i
o
n
3
5
(
2
0
1
2
)
5
1
4
0.35
0.30
0.25
0.20
0.15
0.10
0.05
s
t
a
n
d
a
r
41
Boscalid
Pyraclostrobin
d
mg/kg
air induction nozzle
0.10
0.35
0.08
0.30
0.25
0.06
0.20
0.04
0.15
0.10
0.02
0.08
0.05
0.00
0.00
0.06
0.04
0.02
0.00
'
a
Tetr
ahy
dro
pht
hali
mid
e
mg/kg
0.10
b
b
W
h
Carbendazim
b
M
T
42
Boscalid
Pyraclostrobin
Zone
Fig. 2. Effect of nozzle types (spray quality); Mean values of pesticide residues of boscalid, pyraclostrobin, captan and its degradation compound tetrahydrophthalimide,
carbendazim and thiophanate-methyl from analysis of individual apples picked at four zones within the tree. BI: bottom inside, BU: bottom outside, M: middle, T: top. The
mean
values for all apples are also shown. White bars are standard application Albuz ATR nozzle (fine spray quality); black bars are air induction nozzle application (coarse spray
quality).
Bars labeled with the same letter do not differ significantly p = 0.05.
Northover
et al., 1986). The monograph on captan by FAO/WHO (1970)
also
reports that captan does not appear to undergo any chemical
change on the plant surface. However, Ticha et al. (2007)
showed
captan residues in apples successively decreased until
harvest.
None of the above quoted studies had measured the
degradation
product tetrahydrophthalimide. In this study tetrahydrophthalimide was found in higher levels than captan. The results
showed that approximately 80% of the sum of captan and
tetrahydrophthalimide was found as tetrahydrophthalimide, when
taking the differences in molecular weights into account. The
results are in line with results from Poulsen et al. (2009),
where
7
different spraying strategies were performed in commercial
orchards. However, part of the degradation could occur in the
laboratory during the chemical analysis. Captan is rapidly
degraded
43
Boscalid
Pyraclostrobin
0.00
0.15
0.20
0.25
ME.
0.10Poulsen et al. / Crop Protection 35 (2012) 514
mg/kg
BI BU
standard
T BI BU M
air induction
44
Boscalid
Pyraclostrobin
Standard
mg/kg
0.35
0.30
Captan
0.2
b
5
0.2
0
0.1
5
0.1
- 1
F
Iii
IX
M
BI
BU
air induction
BI
BU
BI BU
standard
T
BI BU M
air induction
45
Boscalid
Pyraclostrobin
BU
mg/kg
M
0.10
standard
Th ioph an ate-methyl
0.08
---------c
0.06
0.04
0.02
BU M
standard
BI BU M
air induction
Carbendazim
bb
Al
0.00 Bl
I
Bl
BU
air induction
46
Boscalid
Pyraclostrobin
Fig. 3. Effect of the zone within the tree; mean values of pesticide residues of boscalid, pyraclostrobin, captan and its degradation compound tetrahydrophthalimide,
thiophanatemethyl and carbendazim, from analysis of individual apples picked at four zones. BI: bottom inside, BU: bottom outside, M: middle, T: top. Results for standard application
(fine spray quality) and air induction nozzle application (coarse spray quality) are shown bars labeled with the same letter do not differ significantly p = 0.05.
-1
47
Table
3
mg/kg
Acceptable Daily Intake (ADI) and Acute Reference Dose (ARfD) in mg kg -1 body
weight for boscalid, pyraclostrobin, captan, tetrahydrophthalimide, carbendazim
and thiophanate-methyl.
Acceptable Daily Intake,
0.04
none
Bupirimate
0.05
0.05
Captan
0.1
Carbendazim
0.02
0.02
Fenoxycarb
0.04
none
Indoxacarb
0.006
0.02
Pirimicarb
0.035
Pyraclostrobin
0.03
Tetrahyd
rophthal
imide
0.3
0.1
0.03
Tetrahydrophthalimide
none found
Thiophanate-methyl
0.08
none found
0.2
Fine sprayquality
Coarse sprayquality
X
Boscali
d nX
mg/k
0.60g
0.30
0.50
0.20
0.40
0.10
0.00
Individual
Captan
Composite
Pyraclostrobin
X
mg/k
0.25g
i
Composit
Individua
0.20
0.15
0.10
0.05
i
I
Composite
Individual
Composite
-----------------------------------------1----------------------------------------------------------------------------,-------------------------------------1
mg/k
1.5 g
0
Fine sprayquality
0.80
0.60
Fine sprayquality
v
1.2
0.7
5
0.5
0
x
1.0
0
I
i
0.25
0.00
Composite
Composite
Individual
1.00
Composite
Individual
Composite
49
Table
3
Thiophanate-methyl
mg/k
0.30g
xx
0.25
0.15
0.10
0.05
Composite
Individual
X
x
0.20
0.15
Composite
Individua
l
0.25
0.20
0.00
Carbendazim
mg/k
0.30g
0.10
i
i
'
0.05
0.00
Composite
Individual
Composite
Individual
Fig. 4. Results for composite and individual samples for pesticide residues of boscalid, pyraclostrobin, captan and its degradation compound tetrahydrophthalimide,
carbendazim
and thiophanate-methyl. Results are shown for both standard application Albuz ATR nozzle (fine spray quality) and air induction nozzle application (coarse spray quality).
50
Table 4
Variability factor for standard and air induction nozzle application for boscalid,
captan,
carbendazim, tetrahydrophthalimide, thiophanate-methyl and pyraclostrobin.
Standard
application
Boscalid
Captan
Carbendazim
Tetrahydrophthalimide
Thiophanate-methyl
Pyraclostrobin
2.0
9.4
1.8
4.0
3.9
2.2
Air induction
nozzle application
2.7
4.3
0.9
2.7
3.8
2.6
The EU MRLs and CODEX MRLs for apples are set for the
mean
residue value of one kg or a minimum of ten apples (European
Commission, 2002; CODEX, 1999). Originally, the toxicological
evaluation of pesticides was performed for chronic effects
only
by
setting an Acceptable Daily Intake (ADI) (Table 3). For control
purposes of ADI an average residue value are more suitable in
order
to smooth out the variation in residue levels, because a
person
eating a high pesticide residue content apple one day will
most
likely eat an apple with at lower content the next day. Consequently, the control for pesticide residues in apples was
decided
to
be carried out for composite samples. Later on, the pesticides
were
evaluated for acute effects and Acute Reference Doses (ARfD)
were
set. To control the ARfDs it is important to know the pesticide
residue in the individual apple, because a person will be
exposed
to
acute effects from a speciic apple, if the residues are high
enough.
However, it would be costly to control the ARfD, if the apple
samples should be analyzed individually as ten apples should
be
analyzed. Instead, the pesticide residue levels in the
composite
samples are calculated back to the assumed highest possible
level
in the individual apples, by applying a variability factor to the
average concentration of ten apples. In EU member states the
variability factor used is seven (EU Commission, 2004),
whereas
CODEX has lowered the factor to a default value of three until
suficient data on single unit is available to calculate a more
realistic
variability factor (WHO, 2010).
To investigate the distribution of the pesticides between
the
apples from the two application modes, all apples were
individually
analyzed. The results of from the analysis of the individual
apples
showed large variations in residue levels, where the apples
with
the
highest concentration had between 12 and 200 times higher
residue
levels than in the apple with the lowest level. To examine the
rela-
Table 5
The minimum and maximum number of individual results not covered by the
variability
factor
3
and
7,
for
boscalid,
pyraclostrobin,
captan,
tetrahydrophthalimid,
carbendazim and thiophanate-methyl, obtained by standard and air induction
nozzle application. The number before the arrows refers to the highest concentration of the 20 mathematically constructed composite samples and the number
after
the arrow refers to the lowest concentration of 20 the mathematically constructed
composite samples. The mathematically constructed composite samples represent
a sample of ten apples used for control of MRLs.
Standard application
Boscalid
Pyraclostrobin
Captan
Tetrahydrophthalimid
Thiophanate-methyl
Carbendazim
Factor 3
0/1
0/1
7 / 24
0/8
2/8
0/0
Factor 7
0/0
0/0
1/4
0/0
0/0
0/0
Air induction
application
Factor 3
1/4
0/0
0/7
0/4
1/6
0/0
Factor 7
0/0
0/0
0/0
0/0
0/1
0/0
51
Table 4
composite
samples was used and one result (the highest) if calculated
with
the
highest result for the composite samples.
As seen from Table 5 the variability factor of seven seems
to
be
adequate to include almost all the results for the pesticides.
For
captan up to 9% of the results were not covered depending on
the
composite concentration chosen. However, the variability
factor
of
three seems to be too low, because up to 30% of the apple
samples
for captan were not covered if the worst case scenario was
chosen.
Also for thiophanate-methyl the factor of three seems to be
too low.
In conclusion, no statistically significant correlation
between
the
fruit weight (size) and residue level was observed, showing
that
the residue levels were independent of the fruit size. None of
the
results for the pesticides residues measured in the apples
exceeded
the EU MRLs. For boscalid and pyraclostrobin no differences
were
seen in the residue levels for the two spraying techniques.
The
coarse spray quality (air induction nozzle) resulted in higher
levels
for captan and thiophanate-methyl at the middle zone and for
tetrahydrophthalimide at the bottom inside zone. However,
with
52
53
54
Zande van de, J.C., Heijne, B., Wenneker, M., 2001. Spray Drift Reduction in
Orchard
Spraying (State-of-the-Art December 2001). Institute of Agricultural and Environmental Engineering/PPO-fruit. IMAG Report 2001-19, Wageningen. 38 p. (in
Dutch with English summary).
Zande van de, J.C., Holterman, H.J., Wenneker, M., May, 2008. Nozzle classification
for drift reduction in orchard spraying: identification of drift reduction class
threshold nozzles. In: Agricultural Engineering International: The CIGR Ejournal
Manuscript ALNARP 08 0014, vol. X.