A.

Neicheva,
Journal of the University D. Technology
of Chemical Karageorgiev,
andD.Metallurgy,
Hadjiev 42, 3, 2007, 307-310

GAS CHROMATOGRAPHIC DETERMINATION OF

MULTI-COMPONENT PESTICIDES IN PLANT PRODUCTS
A. Neicheva1, D. Karageorgiev2, D. Hadjiev1

1
University of Food Technologies, Received 16 May 2007
Plovdiv, 26 Maritza blvd., Accepted 12 August 2007
E-mail: aneicheva@abv.bg
2
Fruit Growing Institute, Plovdiv, 12 Ostromila Str.,
E-mail: dkarageorgiev@abv.bg

ABSTRACT

A fast, selective and precise gas chromatographic method for simultaneous determination of pesticide residues
(chloryiriphos-methyl, cypermethrin, thioziklam, triflumizole and uprodion) in plant products has been developed by
using an electron capture detector and filled capillary columns (2m x 2mm) with 3 % SE-30 and 3 % OV-17 at programmed
change of temperature.
Keywords: analysis, multi-component pesticide residues, GC, fruit plants.

INTRODUCTION included in the technologies for plant protection in our

The contemporary control over the pollution of
fruits and vegetables with pesticide residues is a man-
datory obligation in all countries with advanced agri-
culture and food industry. The methods for its execu-
tion and their improvement are under constant devel-
opment. The accent is put mainly on the fast and easy
access of the analytical procedures, having in mind the
huge number of fresh and processed plant products
reaching consumers on a daily basis. The expert group
Codex Alimentarius of FAO/WHO recommends the new
active substances to be integrated to the existing mul-
tiple methods [1]. On the other side, the exploring of
some newly discovered pesticides in real production
conditions has not been sufficient. The subject of this
research are the insecticides chloryiriphos-methyl,
cypermethrin, thioziklam, triflumizole and uprodion
country which are highly likely to contaminate the crops
as they are applied close to the fruits harvest.
The determination of multiple pesticide residues in
plants and plant products has been conducted most often
with chromatographic methods - TGC, GC and HPLC [1-9].
A lot of controlling bodies (AOACand Codex Alimentarius)
are in the process of developing different analytical schemes
for determination of multi-component pesticides which are
based on different chromatographic techniques. The ma-
jor place in these techniques takes the gas chromato-
graphic method due to its high sensitivity and separat-
ing capability [1, 5-9]. The goal of this research is to
develop an analytical path for simultaneous determination
of residues from three insecticides and two fungicides in
fruit products by using the advantages of the gas chromato-
graphic method which will provide fast and reliable control
over the purity of the fresh and processed plant food.

307
 Journal of the University of Chemical Technology and Metallurgy, 42, 3, 2007
EXPERIMENTAL
Reactives
Standard acetic solutions, containing 0,001 mg/cm3
from the pesticides chloryiriphos-methyl, cypermethrin,
thioziklam, triflumizole and uprodion were used.
Equipment
Gas chromatograph 304 (Pye Unicam) with 63Ni-
capture electron detector, supplied with integrator SP
4270 (Spectra Physics) was used for the research.
Workflow
The explored plant products (apples, peaches,
tomatoes, etc.) are turned into homogenous mixture
from which 50 g are taken. The extract of pesticides is
done by processing of the sample by 130 cm3 methanol
for 40 min on shaking machine. After filtering of that
extract through a silica gel layer, the filtrate is concen-
trated up to 50 cm3 and 150 cm3 10 % potassium chlo-
ride solution is added. The pesticides are re-extracted
two times from the mixture by 80 cm3 column with d=1
cm and 5 g mixed absorbent (potassium sulfite: florisil:
activated charbon = 1:1,5:0,1). The elusion of the pesti-
cides in the column is done with 100 cm3 hexane: ethyl-
ene ether = 95:5. The eluate is evaporated to dry at 40oC
and the dry residue is dissolved in 2 cm3 acetone. 1 ml of
that mixture is chromatographed respectively on columns
with 3 % SE-30 (200 cm x 2mm) at a temperature
programme 150oC (4 min) - 6oC/min - 230oC and with
3 % OV-17 (200 cm x 4mm) temperature programme
220oC - 3oC/min - 240oC. The temperatures of the injec-
tor and detector are 250oC and 300oC, respectively, while
the rate of the carrier gas nitrogen is 40 - 50 ml/min. The
treatment of the chromatograms is done by using the
method of absolute calibration.
RESULTS AND DISCUSSION
Upon the development of an analytical path for
determination of pesticide residues, the fining of ap-
propriate conditions for their effective extraction and
the separation of the substances which disturb their
analysis is of major importance. Acetonitrile and ac-
etone are the most recommended extragents [1]. At the
same time, acetone is preferred as cheaper and more
volatile. As compared with the acetonitrile extracts, the
acetone extracts contain greater amount if refining sub-
308
stances which required investigation for its replacement.
While choosing the proper extragent, it was decided to
use methanol because it is more popular than acetone
and more volatile than acetonitrile, as well as it is ac-
cessible in practice in the controlling laboratories. Af-
ter the conducted experiments, the following order for
changing the effectiveness of the extragents has been
found out: methanol > acetonitrile > acetone.
In order to optimize the extraction process, the
influence of two more factors with significant impor-
tance for the extraction level the time of the process
and the ratio sample:extragent, have been researched.
The results from the conducted research are shown in
Fig. 1.
'
&

%
!
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#
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#
= > ? @
Fig. 1. Contents of residues (in %) of thioziklam (a, c) and
cypermethrin (b, d) in apple extracts, obtained with methanol at
different extraction times (1 - 60 min; 2 - 40 min; 3 - 20 min) and
different ratio sample:extragent ( 4 - 1:3; 5 - 1:2,6 and 6 - 1:1).
Based on the obtained data for the influence of
factors time and volumes of the extragent, the following
optimal conditions have been identified: timing of 40
min and ratio sample:extragent - 1:2,6. The increased
timing of the extraction (60 min) and ratio
sample:extragent (1:3) increase insignificantly the ana-
lytical harvest but at the same time the content of the
accompanying pesticide substances also rises up which
requires their more precise removal.
The refining of the obtained plant extracts from
the extracted disturbing substances (mainly wax and dyes)
has been done by liquid-liquid distribution and follow
up column chromatography. The analytical practice has
shown that a higher effectiveness of the separation has
 A. Neicheva, D. Karageorgiev, D. Hadjiev

been achieved by using solvents with big polarity

difference [1, 3, 5]. After the experiments and having in
mind the polarity of the explored active substances,
hexane for their re-extraction was used. A glass column
with d = 1 cm and 5 g mixed absorbent have been sug-
gested after testing with columns of different size and
contents of absorbents (potassium sulfide: florisil: acti-
vated charbon = 1:1,5:0,1). Taking into account the sol-
vency of the pesticides and their accompanying mix-
tures and after the conducted experiments, the mixed
solvent hexane: ethyl ether (95:5) for elusion of the pes-
ticides from the column has been chosen.
The choice of an appropriate stationary phase is
an important stage in the development of GC method.
The capability of four stable phases for fast separation of
the explored pesticides has been relatively well studied
Fig. 2. Gaschromatogram of refined extract of apples with addition
(SE30, OV101, QF1 and OV-17). Phases SE30 of pesticides (with 0,1 mg/kg):
and OV17 have better characteristics in terms of sen- a on column ( 200 cm x 2 mm) with 3 % SE-30 at temperature
programme 150 oC (4 min) - 6 oC/min - 230 oC [1- thioziklam
sitivity and separation time. Some of the explored (t R = 3.44),2 - chloryiriphos-methyl (tR= 9.30),3 - triflumizole
pesticides have quite different volatility (thioziklam and (tR= 14.12), 4 - uprodion (tR = 16.44)];
cypermethrin), which makes their simultaneous deter- b on column (200 cm x 2 mm) with 3 % OV-17 at temperature
program 220 oC - 3 oC/min - 240 oC [2 - chloryiriphos-methyl
mination impossible on one column and at isothermal (t R= 3.10), 3 - triflumizole (tR= 4.51), 4 - uprodion (tR = 11.08),
temperature regime. 5 - cypermethrin (tR= 28.58)].

Upon continuous chromatography of the filled

capillary columns (200 cm x 2 mm) and appropriately
chosen programs for changes in temperatures, the opti-
mal GC conditions for simultaneous determination of
the explored pesticides has been identified on their com-
mon presence in one sample.
An analytical path for control of products, fro-
zen with residues of the explored pesticides has been
suggested, based on the procedure developed for ex-
traction and refinery of plant extracts, containing the
explored pesticides and the conditions found for their
GC determination. A chromatogram of the refined ex-
tract of apples with additions of the explored pesticides
(0,01 mg/kg) is shown on Fig. 2.
The analytical availability, preciseness and repro-
duction in the GC method have been identified by ana-
lyzing apples based on the method of standard addition
(Table 1). The results obtained show that the analytical
production for most of the pesticides is above 94 % and

Table 1. Gaschromatographic determination of the explored pesticides in apples

using the method of the standard addition.
Name of pesticide Standard addition, Analytical finding, Relative standard
mg/kg % deviation, %
(n=5) (n=5)
chloryiriphos-methyl 0.1 97,2 1,4 2,6

cypermethrin 0.1 90,3 2,5 2,8

thioziklam 0.1 98,5 1,8 3,7

triflumizole 0.1 94,4 3,8 4,1

uprodion 0.1 95,5 3,5 3,7

309
 Journal of the University of Chemical Technology and Metallurgy, 42, 3, 2007
only for cypermethrin is 90 %. Relative error of the
different pesticides vary between 1,4 and 3,8 %, while
the reproduction is from 2,6 to 4,1 %. In addition to
that, low minimum limits of determination (0,002
0,01 mg/kg) have been identified. The method can be
sruccessfully applied in the analysis of other plants
(peaches, plums, tomatoes, cucumbers, etc.) both in their
fresh and processed state.
CONCLUSIONS
A fast, selective and easily accessible analytical
path for determination of residues from three insecti-
cides and two fungicides in fruit products has been de-
veloped by using the advantages of the gas chromato-
graphic method. The analytical characteristics of the
method (sensitivity, preciseness and reproducibility of
the measurements) show that it is appropriate for con-
trolling residues of the explored pesticides in fresh and
processed plant products. The same method can be ap-
plied for analyzing of residues from similar pesticide
compounds.
REFERENCES
1. A. Ambrus, E. Visi, F. Zakar, E. Hargital, L. Szabo,
A.Papa, General method for determination of pesti-
cides residues in samples of plant origin, soil anal wa-
ter, J. Assoc. of Anal. Chem., 64,1986, 1035 1052.
2. K.K. Singh, M.S. Shekhawat, Thin Layer Chromato
graphic Methods for Analysis of Pesticide Residues
310
in Environmental Samples, Journal of Planar Chro-
matography, 11, 1998, 64-185.
3. G. Kempe, U.Schumann, K.Speer, Determination of
pesticides in foods of vegetal origin by thin-layer
chromatography, Deutsche Lebensmittel Rundschau,
'#, 6, 1999, 231-234.
4. C. Trova, O. Zerbinati, L. Badan, M.G. Gnazzotti,
HPLC determination of pesticide residues in veg-
etables (fruits) by means of an ethyl acetate/n-hex-
ane solvent system, Judustrie-Alimetary, 38, 385,
1999, 1090-1093.
5. F. Kovacs, Multi-residue method for pesticide residue
analysis of vegetable and fruit samples of high water
content, Novenyvedelem, !$, 8, 2000, 397-403.
6. E.Ueno, H.Oshima, I.Saito, H.Matsumoto,
Multiresidue analysis of organophosphorus pesti-
cides in vegetables and fruits using dual-column GC-
FPD-NPD, Journal of the Food Hygienic Society of
Japan, " , 6, 2001, 385-393.
7. M. Gamon, G.Leo, A.Ten, F.Mocholi, Multiresidue
determination of pesticides in fruit and vegetables
by gas chromatography/tandem mass spectometry,
Journal of AOAC International, &", 4, 2001, 1209-
1216.
8. H.Kobayashi, The approach to pesticide performance
in analytical procedure, Journal of the Food Hy-
gienic Society of Japan, " , 6, 2002, J331-J336.
9. W. Schwack, M. Anastassides, E. Scherbaum, Multi-
methoden versus Wirkstoffvielfalt Rchstandsanalytik
von Pflauzenschutzmitteln, Chem. Unseres Zeit., 37,
2003, 324-333.