Environ Geochem Health (2007) 29:45–50

DOI 10.1007/s10653-006-9060-2

ORIGINAL PAPER

Imidacloprid residues in fruits, vegetables and water

samples from Palestine
Ayman Daraghmeh Æ Amjad Shraim Æ
Sameer Abulhaj Æ Ramzi Sansour Æ
Jack C. Ng

Received: 4 June 2005 / Accepted: 22 August 2006 / Published online: 25 November 2006

Springer Science+Business Media B.V. 2006

Abstract The aim of this work was to report on
imidacloprid [IUPAC name 1-(6-chloro-3-
pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine]
residues in some vegetables, fruits, and water
samples collected from the West Bank, Palestine,
in 1998 and 1999. Imidacloprid and its derivatives
in the study samples were extracted by methanol/
water and oxidized into 6-chloronicotinic acid and
subsequently derivatized into 6-chloronicotinic
acid trimethylsilyl ester before being determined
by GC/MS. Imidacloprid residues were detected
in more than half of the analyzed samples. The
highest and lowest imidacloprid concentrations
were found in eggplant (0.46 mg/kg) and green
beans (0.08 mg/kg), respectively. An increase of
11–120% in imidacloprid concentration in the
1999 samples was observed when compared with
those of 1998. This may suggest imidacloprid
accumulation in the soil and/or increased use by
local farmers. The imidacloprid residue
concentrations in several crops were found to
exceed the CODEX maximum residue limit.
Keywords Pesticides Æ 6-Chloronicotinic acid Æ
1-(6-Chloro-3-pyridylmethyl)-N-nitroimidazolidin-
2-ylideneamine Æ West Bank

A. Daraghmeh
Central Public Health Laboratory, Ramallah
West Bank, Palestine
A. Shraim
Chemistry Department, Faculty of Science,
Taibah University, P. O. Box 344, Almadinah
Almunawwarah, Kingdom of Saudi Arabia
A. Shraim (&) Æ J. C. Ng
National Research Centre for Environmental
Toxicology, The University of Queensland,
39 Kessels Road, Coopers Plains, Brisbane,
Queensland 4108, Australia
e-mail: a.shraim@uq.edu.au
S. Abulhaj Æ R. Sansour
The Centre of Environmental Health,
Berzeit University, Berzeit, West Bank, Palestine
Introduction
Imidacloprid [IUPAC name 1-(6-chloro-3-
pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine]
is a chloronicotinyl insecticide first introduced by
Bayer Agricultural Products (Leverkusen,
Germany). It was commercially developed and
marketed in the early to mid-1990s. Imidacloprid
is a general-use pesticide used worldwide for
crop-, fruit-, and vegetable pest control, termite
control, and flea control in dogs and cats (Ishii
et al., 1994; Cox, 2001). It is most commonly used
on rice, cereal, maize, potatoes, vegetables, sugar
beet, fruit, cotton, hops, and turf (Heijbroek &
Huijbregts, 1995; Huijbregts, Gijssel, &

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Heijbroek, 1995; Ahmed, Kanan, Inanaga, Ma, &
Sugimoto, 2001; Dikshit & Lal, 2002; Kuhar,
Stivers-Young, Hoffmann, & Taylor, 2002).
Imidacloprid is the active ingredient in a wide
variety of products, such as Gaucho, Alcador, and
Confidor. It is a relatively new type of insecticide,
possessing potential activity and novel modes of
action (Moriya et al., 1992; Moffat, 1993;
MacDonald & Meyer, 1998).
Imidacloprid degrades in soil and plants into a
number of different derivatives, all of which
contain the chloropyridine structure with the
6-chloronicotinic acid as a major metabolite
(Ishii et al., 1994; de Erenchun, Gomez de
Balugera, Goicolea, & Barrio, 1997). The half-
life of the insecticide in soil is reported to range
from 29–190 days (Sarkar, Roy, Kole, &
Chowdhury, 2001). On the other hand, Cox
(2001) reported that imidacloprid is persistent in
soil, as the concentration of tested soil samples
had not decreased 1 year after application. It is
now accepted that successive treatments in soil
can lead to its accumulation in soil and crops
(Ishii et al., 1994; de Erenchun et al., 1997).
Moreover, owing to its persistence (Cox, 2001;
Sarkar et al., 2001), high solubility and mobility,
imidacloprid has the potential to leach into the
groundwater (Cox, 2001). According to Cox,
Koskinen and Yen (1997), the solubility of
imidacloprid in water was 500 mg/l, whereas
the partition coefficient (Koc) for imidacloprid
was found to be dependent on the initial solution
concentration and type of soil [Koc 71–81 (initial
concentration 250 mg/l), 415–531 (initial
concentration 1.5 mg/l) and 802–1560 (initial
concentration 0.05 mg/l)].
Imidacloprid is classified by the US
Environmental Protection Agency (US EPA) as
both a toxic class II and a class III agent, and
must be labeled with the signal word ‘‘Warning’’
or ‘‘Caution’’. The acute and chronic reference
doses for imidacloprid for all population
subgroups [including infants ( < 1 year) and
children (1–12 years)] as set by the US EPA
(1999) are 0.42 mg/kg per day and 0.057 mg/kg
per day, respectively. Imidacloprid was also
reported to be very toxic to aquatic
invertebrates, toxic to upland game birds, and of
low toxicity to fish (Cox, 2001).
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Environ Geochem Health (2007) 29:45–50
In Palestine, imidacloprid (as Gaucho) has
been used for agricultural purposes since the
1980s. The average use of commonly used
agrochemicals, including imidacloprid, per
growing season in Palestine was estimated to be
65.5 kg/ha for greenhouse tomatoes, 38.7 kg/ha
for greenhouse cucumbers, and 74.6 kg/ha for
open-field cucumbers. Owing to the local political
situation, imidacloprid residues in Palestinian
crops have never been determined before.
Therefore, the objective of this study was to
determine the residues of imidacloprid (parent
compound) and its derivatives (after oxidation to
6-chloronicotinic acid) in some local crops as they
reach the consumers. For this purpose, samples of
various food crops were collected from the local
markets in the West Bank, Palestine and analyzed
by GC/MS. In order to check if imidacloprid
residues have contaminated the water resources,
we also collected and analyzed groundwater
samples from an extensive area ofagricultural
activity.
Experimental method
Chemicals
6-Chloronicotinic acid (99%), sodium hydroxide
(GR 97%), anhydrous sodium sulfate (99%),
potassium permanganate (GR 99%), sodium
bisulfite (ACS reagent grade), t-butylmethyl
ether (99%), and Amberlite XAD-4 (20–60
mesh) were all obtained from Sigma–Aldrich,
Germany. Acetonitrile and methanol (both HPLC
grade) were obtained from Merck, Germany. N-
methyltrimethylsilyltrifluoroacetamide (MSTFA)
was obtained from Alltech, USA. Analytical
standard imidacloprid was a gift from Bayer,
Germany. Nitrogen gas (99.999%) was obtained
locally. All deionized water used was organic-free
Milli-Q water (Millipore, USA).
Equipment
A gas chromatograph (GC 17A, Shimadzu, Japan)
equipped with an auto-injector and a mass
spectrometer detector (QP 5000 MSD) in the
Environ Geochem Health (2007) 29:45–50
single ion-monitoring (SIM) mode was used. A
DB-5 capillary column (5% phenylmethyl
polysiloxane polymers), 30 m · 0.25 mm · 0.25
lm from J & W Scientific (Folsom CA, USA) was
utilized. The carrier gas was helium at 100 kPa and
flow rate of 1 ml/min. The injector temperature
was 260C. The oven temperature was
programmed as follows: initial temperature
100C, held for 1 min, ramped at 15C/min to
180C, then at 30C/min to 300C, and held for
3 min. The detector was switched on 3 min post-
injection after the elution of the solvent at the
beginning of the temperature program. The two
ions used in the quantification of 6-chloronicotinic
acid–trimethylsilyl ester were 214 as the target ion
and 170 as the qualifying ion.
Sample collection
Eleven types of vegetables and fruits (apples,
bananas, cauliflower, cucumber, eggplant, grapes,
green beans, maize, peaches, potatoes, and
watermelon) were purchased from the local
markets of the West Bank, Palestine during
1998–1999. At the same time, 19 groundwater
samples were collected from the extensive
agricultural area around Jenin city.
Extraction and analysis of imidacloprid
Imidacloprid and its derivatives in the samples
were extracted and oxidized to 6-chloronicotinic
acid as per the method of Placke & Weber
1993. Briefly, approximately 1 kg of each
vegetable or fruit sample was chopped into
small pieces and made into a slurry. A
representative portion of the slurry (50 g) was
then taken and soaked in methanol:water (3:1
v:v, 300 ml) for 30 min. The mixture was then
homogenized in a blender and filtered, and the
filtrate volume was made up to 500 ml with
methanol. A portion of this solution (100 ml)
was concentrated to approximately 20 ml in a
rotary evaporator at 60C and then transferred
to a pre-conditioned column packed with XAD-
4 resin (for water samples, an aliquot of 250 ml
was concentrated as above and transferred to
the column). The column was flushed twice with
methanol:water (3:1 v:v, 20 ml), then the
47
retained compounds were eluted with
methanol (100 ml) and concentrated to 1 ml in
a vacuum rotary evaporator and gentle stream
of nitrogen. Imidacloprid and other derivatives
containing the 6-chloropicolyl moieties extracted
in the 1 ml concentrate were oxidized to 6-
chloronicotinic acid as follows: the concentrate
was diluted in water (100 ml) before we added
the oxidizing solution [sodium hydroxide (32%,
5 ml) and potassium permanganate (5%,
50 ml—100 ml added to samples of hops)].
The mixture was then quickly refluxed with
stirring for 5 min. Another 50 ml water was
added. The flask was placed in an ice bath and
allowed to cool to 15C under agitation for
10 min. Sulfuric acid (10%, 50 ml) was then
added, followed by three additions of solid
sodium bisulfite (approx 1 g each addition)
under cooling and agitation. More sulfuric acid
was added, when necessary, to maintain a pH
value of £1. Subsequently, the solution was
extracted with t-butylmethyl ether (3 · 50 ml).
The organic phase was filtered through
anhydrous sodium sulfate (30 g) and
evaporated to dryness in a rotary evaporator.
The extract was then dissolved in acetonitrile
(2 ml). An aliquot (250 ll) of this solution was
derivatized to 6-chloronicotinic acid-
trimethylsilyl ester by mixing it vigorously with
MSTFA (250 ll). An aliquot (1 ll) was then
injected into the GC/MS under splitless mode.
Standard solutions of 6-chloronicotinic acid
were derivatized in the same way as above,
and their aliquots were injected prior to those
of the samples. The analytical method, as used
in this work, was originally validated by Placke
& Weber 1993. However, some of the quality
control parameters were re-checked and found
to be similar to the findings of Placke and
Weber 1993; the method detection level (MDL)
(calculated from three standard deviations of a
blank treated by the same procedure as above)
was found to be 0.03 mg/kg, and the spike
recovery was better than 75%. The standard
deviation of three repeated injections of the
extract of cucumber (1998) was 0.04 mg/kg,
whereas the standard deviation of three
repeated analyses of eggplant (1998) was
found to be 0.05 mg/kg.
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Results and discussion
The chemical structures of imidacloprid,
6-chloronicotinic acid, and 6-chloronicotinic
acid-trimethylsilyl ester are shown in Fig. 1.
Under the above experimental conditions,
6-chloronicotinic acid-trimethyl silyl ester elutes
at around 12 min. The concentrations of residues
of imidacloprid in fruits, vegetables, and water
samples are summarized in Table 1. The highest
average concentration was found in the eggplant
(0.41 mg/kg), followed by potatoes (0.40 mg/kg),
peaches (0.36 mg/kg), and watermelons (0.32 mg/
kg). On the other hand, the lowest concentration
was found in grapes (0.08 mg/kg), followed by
green beans (0.10 mg/kg) and cucumber (0.11 mg/
kg). Although collected from an extensive area
ofagricultural activity, the water samples were
found to have imidacloprid concentrations below
the MDL (0.03 mg/kg). According to a report by
the US EPA 1997, imidacloprid residues were not
detected in any wells, ponds, lakes, streams, etc.
near sites where imidacloprid was applied.
However, this situation may change, as other
groundwater monitoring studies for imidacloprid
residues are currently underway in several parts
of the USA.
Fig. 1 Chemical structure of imidacloprid, 6-chloronicot-
inic acid, and 6-chloronicotinic acid trimethylsilylester
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Environ Geochem Health (2007) 29:45–50
Imidacloprid residues were found in 66% and
73% of the samples collected in 1998 and 1999,
respectively. Of the 122 samples collected in the
2-year study, more than 90% were of domestic
origin, whereas the rest were imported from
Israel and/or were of unknown origin.
Imidacloprid residues for most of the samples
collected in 1999 were greater by 11–120% when
compared with those of 1998. This increase may
be explained by accumulation of imidacloprid in
the soil and/or more quantities per hectare of the
insecticide have been applied in 1999. However,
larger scale and more in-depth research is needed
before an accurate assessment of the situation of
this compound can be made.
Imidacloprid was reported to be persistent in
soil (decay time for 50% of imidacloprid (DT50)
is 2 years), with a high potential for carry-over
and build-up of chemical residues (Sandler, 2001).
A study on crops grown in soils treated with
imidacloprid 1 year and 2 years before, but not
during the year when the investigation was
carried out, showed that the average
concentration of imidacloprid in crops grown on
soils 1 year before was 4.8 lg/kg compared with
8.6 lg/kg for crops grown on soils treated with
imidacloprid for two consecutive years
[Coordination des Apiculteurs de France (CAF,
2000)].
The CODEX 2005), Australian [Australian
Pesticides and Veterinary Medicines Authority
(APVMA, 2006)] and Canadian [Pest
Management Regulatory Agency (PMRA,
2005)] maximum residue limits (MRLs) in most
of the samples analyzed for this study are listed in
Table 2. The highest imidacloprid residue
concentration detected in either year in bananas
was greater than the CODEX MRL (no MRL set
for Australia and Canada); in eggplant it was
greater than the CODEX and Canadian MRLs
(but not more than the Australian MRL); in
maize it was greater than the Australian and
Canadian MRLs (no MRL in CODEX); in
potatoes it was greater than the Canadian MRL
(less than CODEX and Australian MRLs), and,
in watermelon, it was greater than the CODEX
and Australian MRLs (no MRL set for Canada).
Little work has been published in the literature
on the contamination of food crops by
Environ Geochem Health (2007) 29:45–50 49

Table 1 Residues of imidacloprid in Palestinian crops and water samples collected in 1998 and 1999
Product No. of samples No. of samples Percentage of Imidacloprid residue (mg/kg)
analyzed with imidaclo- samples with
prid residue imidacloprid
residue
a
1998 1999 1998 1999 1998 1999 Average 1998 1999

Apples 5 6 5 6 100 100 0.24 0.23 0.26

Bananas 3 5 1 3 33 60 0.18 0.16 0.21
Cauliflower 5 5 4 4 80 80 0.26 0.30 0.19
Cucumber 5 5 4 5 80 100 0.11b 0.13 0.09
Eggplant 3 6 3 6 100 100 0.41c 0.36 0.46
Grapes 4 5 4 4 100 80 0.08 0.05 0.11
Green beans 5 3 3 3 60 100 0.10 0.12 0.08
Maize 4 2 3 2 75 100 0.28 0.27 0.30
Peaches 4 4 2 2 50 50 0.36 0.32 0.41
Potatoes 7 5 7 5 100 100 0.40 0.38 0.43
Watermelon 6 6 3 4 50 75 0.32 0.33 0.32
Water 8 11 0 0 0 0 < DLd < DLd < DLd
Total 59 63 39 46 66 73
a
Average value of imidacloprid residue (mg/kg) of all the samples collected during both years
b
Standard deviation of repeated injection is 0.04 mg/kg (n=3).
c
Standard deviation of repeated analysis is 0.05 mg/kg (n=3)
d
Below method detection level, which is 0.03 mg/kg

Table 2 The CODEX (2005), Australian (APVMA 2006), and Canadian (PMRA 2005) imidacloprid maximum residue
limits (in milligrams per kilogram) in crops analyzed for this study
Food Item This study, maximum concentration CODEX MRL Australian MRL Canadian MRL

Apple 0.26 0.5 0.3 0.5

Banana 0.21 0.05 _ –
Cauliflower 0.30 0.5 0.5 3.5
Cucumber 0.13 1.0 0.2 0.5
Eggplant 0.46 0.2 0.5 0.08
Grapes 0.11 1.0 0.1 1.5
Green beans 0.12 2.0 – –
Maize 0.30 – 0.05 0.05
Peaches 0.41 0.5 0.5 –
Potatoes 0.43 0.5 0.5 0.3
Watermelon 0.33 0.2 0.2 –

imidacloprid, probably because this insecticide is authors (Fernandez-Alba, Tejedor, Aguera,

not included in the food monitoring programs of
most countries (Cox, 2001). Fernandez-Alba,
Valverde, Aguera, Contreras, & Chiron (1996)
found residues of imidacloprid in the range of
0.01–0.3 mg/kg in all analyzed samples (45
samples of peppers, tomatoes, and cucumbers)
tested one week after treatment. In a crop-
monitoring program, where 200 samples of fruits
and vegetables were analyzed for pesticide
residues, including imidacloprid, the same
Contreras, & Garrido, 2000) found imidacloprid
residues in 25–53% (average 21%) of the 200
samples analyzed. Only two tomato samples out
of the total of 42 samples that contained
imidacloprid residues were found to exceed the
limit set by the Spanish government.
The findings of this study are worrying.
Therefore, it is highly recommended that a
monitoring program for imidacloprid residues in
food crops at the national level be initiated.

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Acknowledgments The authors thank both Dr. Aqel
Abu Qaree and Dr. Aarif El-Mubarak for their technical
support. The project was funded by NIEHS under the
MERC program. EnTox is jointly funded by Queensland
Health, Griffith University, Queensland University of
Technology, and The University of Queensland, Australia.
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