Application Note

Food Testing and

Agriculture

Analysis of Organophosphorus and

Organochlorine Pesticides in Fruit and
Vegetables Using an Agilent 8890 GC
with Four Detectors

Author Abstract
Youjuan Zhang
Agilent Technologies
This Application Note describes an effective and reliable analytical method for
(Shanghai) Co. Ltd,
the determination of organophosphorus and organochlorine pesticide residues in
Shanghai 200131 P.R. China
fruit and vegetables following the China Standard NY/T 761-2008.1 An unpurged
two-way capillary flow technology (CFT) device was used to split the sample 1:1
onto two columns then to two detectors. Compared with the traditional pretreatment
procedure described in NY/T 761-2008, a simplified Quick, Easy, Cheap, Effective,
Rugged, and Safe (QuEChERS) method was used in this application, and provided
sufficient sample matrix cleanup while preserving low-level analyte detection. Area
repeatability, linearity, and recovery were evaluated for both organophosphorus and
organochlorine pesticides using an Agilent 8890 GC system with four detectors.
Introduction
The use of pesticides has played an
important role in the prevention and
control of pests and the resulting
improvement of agricultural yields.
Two common pesticide classes
used in agricultural treatments have
organophosphorus and organochlorine
chemical structures.
For the determination of
organophosphorus and organochlorine
pesticide residues in fruit and
vegetables, gas chromatography (GC),
gas chromatography/mass
spectrometry (GC/MSD), and gas
chromatography/tandem mass
spectrometry (GC/QQQ) methods are
widely used. Correspondingly, China
has issued a series of standards for
the determination of those pesticides.
NY/T 761-2008 describes a GC method
with an electron capture detector
(ECD) and flame photometric detector
(FPD). GB/T 19648-20062 describes a
GC/MSD method for 500 pesticides,
and GB 23200.113-20183 is a GC/QQQ
method for 208 pesticides. Mass
2
spectrometry methods have obvious
advantages in qualitative analysis, and
can determine dozens or even hundreds
of pesticide residues simultaneously with
high efficiency. Consequently, the price
of the instrument is relatively expensive.
Although gas chromatography has
less qualitative ability than MS, it is still
welcomed by many labs because of its
selective detectors and low detection
cost. ECD has excellent selectivity for
chlorine, and is a good choice for the
analysis of organochlorine pesticides.
FPD has high selectivity for sulfur and
phosphorus, and it is a good choice
for the analysis of organophosphorus
pesticides. In the NY/T 761 method,
a primary column and confirmation
column are used in tandem to achieve
accurate qualitative analysis and prevent
false-positive results. This dual-column
approach also provides quantitative
results for both organophosphorus
and organochlorine pesticides when
used with two FPDs and two ECDs.
Normally, labs need two GC systems
to follow the NY/T 761 method strictly.
However, the 8890 GC can have two
FPDs and two ECDs installed on one
instrument to provide greater flexibility
compared to other lab GCs. For both
organophosphorus and organochlorine
pesticides analysis, labs do not need
to change hardware; the only thing
they need to do is to reinstall different
columns onto different detectors.
Sample pretreatment is important
for the analysis of multipesticide
residues, which directly affects the work
efficiency and sensitivity. The NY/T 761
method uses traditional, labor-intensive
extraction and cleanup procedures
for sample pretreatment. The cleanup
procedure of organochlorine pesticides
and organophosphorus pesticides is
different. That means two different
pretreatments are needed for the same
sample when organophosphorus and
organochlorine pesticides both need
to be tested. The popular sample
pretreatment method of QuEChERS is
the optimal method for high-throughput
sample analysis. Most importantly, the
same sample pretreatment process can
be used both for organophosphorus and
organochlorine pesticides, which greatly
improves the analysis efficiency.
Experimental A FPD1 B ECD1

FPD2 ECD2

Instrumentation HP-50+
HP-1
DB-5
DB-17
Inlet Inlet
An 8890 GC with an SSL inlet equipped
with two ECDs and two FPDs was Splitter Splitter
used for this series of experiments. An Retention Retention
gap gap
unpurged two-way CFT device was used
to split the sample 1:1 onto two columns Figure 1. Agilent capillary flow technology two-way splitter without makeup gas (p/n G3181B) and diagram
of instrument setup of simultaneous confirmation from a single injection onto both the primary and
for detection on two detectors. For confirmation columns. (A) FPDs for phosphorus detection, (B) ECDs for chlorine detection.
organophosphorus pesticides analysis,
the primary column and confirmation
column were Agilent HP-50+ and Table 1. Chromatographic conditions.

Agilent HP-1. A dual-column, dual-ECD Organophosphorus method

system with an Agilent DB-5 primary
analysis column and an Agilent DB‑17
confirmatory column was used to
Inlet
separate the organochlorine pesticides.
The analysis on organophosphorus
pesticides and organochlorine pesticides Liner
GC Agilent 8890 GC equipped with two FPDs
Split/splitless
Temperature: 220 °C
Splitless mode, purge flow 60 mL/min at 0.75 min.
Agilent Ultra Inert, splitless, single taper, glass wool (p/n 5190-2293)

analysis cannot be run simultaneously Injection 2 µL

because of their different temperature Retention Gap 0.5 m × 0.53 mm id deactivated fused silica tubing (p/n 160-2535-5)
programs. Compared with the Column
Column1: Agilent HP-50+ 30 m × 0.53 mm, 1 µm (p/n 19095L-023)
Column2: Agilent HP-1 30 m × 0.53 mm, 1.5 µm (p/n 19095Z-323)
Agilent 7890 GC, which can only install
Carrier Nitrogen, 10 mL/min, constant flow (the column flow is the same for column 1 and 2)
three detectors at most, the 8890 GC
is more flexible, and can install four Oven 150 °C (2 minutes), 8 °C/min to 250 °C (12 minutes)
Temperature: 250 °C
detectors at the same time. Therefore, Emission block: 150 °C
labs do not need to change the hardware; FPD Plus 1 and 2 Hydrogen: 60 mL/min
Air: 60 mL/min
they must only reinstall the appropriate Make-up gas (N2): 60 mL/min
columns to the right detectors. Figure 1
Organochlorine method
shows the schematics for the instrument
GC Agilent 8890 GC equipped with two ECDs
setup. Table 1 lists the chromatographic
conditions used for these analyses. Split/splitless
Inlet Temperature: 200 °C
Reagents and chemicals Split mode, split ratio: 10:1

All reagents and solvents were HPLC Liner Agilent Ultra Inert, split, low pressure drop, glass wool (p/n 5190-2295)
grade. Acetonitrile (ACN) and hexane Injection 2 µL
were purchased from J&K Scientific Retention Gap 0.5 m × 0.53 mm id deactivated fused silica tubing (p/n 160-2535-5)
LTD. Acetone was purchased from Column1: Agilent DB-5 30 m × 0.25 mm, 0.25 µm (p/n 122-5032)
Column
ANPEL Laboratory Technologies Column2: Agilent DB-17 30 m × 0.25 mm, 0.25 µm (p/n 122-1732)

(Shanghai) Inc. All organophosphorus Carrier Nitrogen, 1 mL/min, constant flow (the column flow is the same for column 1 and 2)

and organochlorine single standards Oven

were purchased from J&K Scientific LTD
and ANPEL Laboratory Technologies
(Shanghai) Inc.
150 °C (2 minutes), 6 °C/min to 270 °C
(12 minutes, hold 23 minutes for deltamethrin analysis)
Temperature: 320 °C
ECD 1 and 2
Make-up gas (N2): 25 mL/min

3
 Table 2. Data results for organophosphorus pesticides analysis on an Agilent HP-50+ column.
Solutions and standards
Table 2 shows 54 organophosphorus Linearity % RSD (n = 8)
pesticides divided into four groups. Range MDL
No. Name (mg/kg) R2 Low Middle High (mg/kg) % Recovery Group
According to the response values of
1 Dichlorvos 0.05–0.5 0.9982 2.8 1.5 1.5 0.004 108.9 1
each pesticide on the instrument, a
2 Acephate 0.05–0.5 0.9989 4.6 3.1 1.6 0.007 97.7 1
certain volume of single pesticide
3 Dicrotophos 0.05–0.5 0.997 4.3 2.3 1.6 0.007 105.3 1
standard solution of the same group
4 Disulfoton 0.05–0.5 0.9984 3.1 2.7 2.1 0.006 118.2 1
was accurately added and diluted with
5 Dimethoate 0.05–0.5 0.9981 0.6 1.6 1.3 0.002 111.4 1
acetone. The same method was used to
6 Parathion-methyl 0.05–0.5 0.9984 1.6 2.1 1.6 0.003 116 1
prepare four groups of stock solutions
7 Chlorpyrifos 0.05–0.5 0.9982 2.7 1.7 1.3 0.003 115.1 1
of organophosphorus pesticide mixture.
8 Pirimiphos-ethyl 0.05–0.5 0.9985 2.6 1.4 1.4 0.003 111.8 1
Calibration standards were diluted by
matrix blank (see Sample preparation). 9 Fenthion 0.05–0.5 0.999 3 2.4 1.6 0.005 111 1

10 Phoxim 0.2–2.0 0.9922 4.4 2.8 3.6 0.05 110.9 1

11 Ditalimfos 0.05–0.5 0.9994 2.6 1.6 1.1 0.004 70.4 1

12 Triazophos 0.05–0.5 0.9992 3.6 2 2.4 0.007 104.5 1

13 Phosmet 0.2–2.0 0.9998 2.2 2.2 1.6 0.009 102 1

14 Trichlorfon 0.2–2.0 0.999 3 3.5 2.2 0.05 115.4 2

15 Ethoprophos 0.05–0.5 0.9987 1.2 1.5 1.9 0.004 98.5 2

16 Phorate 0.05–0.5 0.9988 2 1.2 1.9 0.004 97.4 2

17 Omethoate 0.05–0.5 0.9982 4.5 3.7 1.8 0.008 102.5 2

18 Diazinon 0.05–0.5 0.998 2.5 1.5 1.9 0.006 95 2

19 Fonofos 0.05–0.5 0.9968 3.5 2.3 2.2 0.003 87.4 2

20 Chlorpyrifos-methyl 0.05–0.5 0.9986 2.4 1.2 1.7 0.004 92.1 2

21 Paraoxon 0.05–0.5 0.9991 3.5 2 1.1 0.007 97.2 2

22 Fenitrothion 0.05–0.5 0.9992 2.9 2.8 1.4 0.005 97.4 2

23 Bromophos 0.05–0.5 0.9986 3.6 3.1 1.1 0.009 100.2 2

24 Bromophos-ethyl 0.05–0.5 0.999 2.2 1.6 0.9 0.007 101 2

25 Profenofos 0.05–0.5 0.9995 3.4 2.7 0.8 0.008 104.7 2

26 Ethion 0.05–0.5 0.9995 1.6 1.7 0.8 0.004 111.6 2

27 Pyrazophos 0.2–2.0 0.9998 2.4 3.1 2.4 0.02 108.6 2

28 Coumaphos 0.2–2.0 0.9997 4.2 2.6 2.6 0.02 107.1 2

29 Methamidophos 0.05–0.5 0.9999 3.5 3.2 2.9 0.004 107.7 3

30 Sulfotep 0.05–0.5 0.9999 2.1 1.2 1.7 0.001 94.1 3

31 Terbufos 0.05–0.5 0.9999 2.2 2.1 2.2 0.003 94 3

32 Monocrotophos 0.05–0.5 0.9995 3.2 0.8 1.6 0.004 93.4 3

33 Dichlofenthion 0.05–0.5 0.9999 3.4 1.7 1.4 0.003 93.2 3

34 Fenchlorphos 0.05–0.5 0.9999 2.2 1.4 1.5 0.003 94.7 3

35 Pirimiphos-methyl 0.05–0.5 0.9999 2.8 2 1.5 0.004 94.2 3

36 Parathion 0.05–0.5 0.9997 3.2 1.5 1.3 0.003 93.9 3

37 Isofenphos 0.05–0.5 0.9999 3.9 3 2.1 0.005 93.5 3

38 Methidathion 0.05–0.5 0.9998 2.7 1.7 1.3 0.004 93.6 3

39 Phosfolan-methyl 0.05–0.5 0.998 2.3 2.9 1.7 0.01 102 3

40 Famphur 0.05–0.5 0.9999 2.7 2.6 3.2 0.02 102 3

41 Phosalone 0.2–2.0 0.9993 2.9 3.1 2.3 0.008 102 3

42 Azinphos-ethyl 0.2–2.0 0.9996 2.8 2.3 1.7 0.02 116.5 3

43 Naled 0.1–1.0 0.9999 2.6 3.3 1.9 0.02 95.5 4

44 Mevinphos 0.05–0.5 0.9998 3.9 2.6 1.3 0.005 118.1 4

45 Propetamphos 0.05–0.5 0.9995 4 2.8 1.4 0.007 101.5 4

4
Table 3 shows 41 organochlorine Table 2. Data results for organophosphorus pesticides analysis on an Agilent HP-50+ column (continued).

pesticides divided into three groups. Linearity % RSD (n = 8)

A certain volume of single pesticide Range MDL %
No. Name (mg/kg) R2 Low Middle High (mg/kg) Recovery Group
standard solution of the same group
was accurately added and diluted with 46-1 Phosphamidon-1 4
0.1–1.0 0.9999 3.6 1.9 0.9 0.02 100.7
hexane. The same method was used to 46-2 Phosphamidon-2 4

prepare three groups of stock solutions 47 Trichloronate 0.05–0.5 0.9999 2.3 2.7 1.1 0.004 97.5 4

of organochlorine pesticide mixture. 48 Malathion 0.05–0.5 0.9999 1.9 1.9 1 0.005 98.1 4

Calibration standards were diluted by 49 Isocarbophos 0.05–0.5 0.9999 2.6 1.8 1 0.004 96.9 4

matrix blank (see Sample preparation). 50 Quinalphos 0.05–0.5 0.9999 2.8 1.6 1 0.004 97 4

51 Tetrachlorvinphos 0.05–0.5 0.9998 2.1 2.4 0.8 0.007 97.9 4

52 Phosfolan 0.05–0.5 0.9999 2.7 3.3 2.5 0.02 97.2 4

53 EPN 0.05–0.5 0.9993 2.8 3.8 1.8 0.009 100.4 4

54 Azinphos-methyl 0.2–2.0 0.9995 2 3.7 1.5 0.02 91.3 4

Table 3. Data results for organochlorine pesticides analysis on an Agilent DB-5 column.

% RSD (n = 8)
Linearity
Range 0.05 0.1 0.5 MDL %
No. Name (mg/kg) R2 mg/kg mg/kg mg/kg (mg/kg) Recovery Group
1 α-BHC 0.05–0.5 0.9996 1.1 1.2 1 0.00003 98.6 1

2 Simazine 0.05–0.5 0.9931 0.8 1.3 1 0.002 88.1 1

3 Atrazine 0.05–0.5 0.9912 2.1 0.9 1 0.002 78.3 1

4 δ-BHC 0.05–0.5 0.9991 0.6 1.3 1 0.00003 93.7 1

5 Heptachlor 0.05–0.5 0.9996 0.8 1.1 0.8 0.00003 118.6 1

6 Aldrin 0.05–0.5 0.9997 0.6 1.2 0.8 0.00004 108.5 1

7 o,p’-DDE 0.05–0.5 0.9997 1.1 1.2 0.7 0.00004 101.7 1

8 p,p’-DDE 0.05–0.5 0.9998 0.7 1.1 0.9 0.00005 106.6 1

9 o,p’-DDD 0.05–0.5 0.9996 1.2 1.1 0.8 0.00004 77.3 1

10 p,p’-DDT 0.05–0.5 0.9997 0.9 0.5 0.5 0.00006 111 1

11 Iprodione 0.05–0.5 0.9974 1 1.3 1.7 0.0007 113.5 1

12 Bifenthrin 0.05–0.5 0.9998 1 1.7 0.8 0.0002 116 1

13 cis-Permethrin 0.05–0.5 0.9999 1.6 2.2 0.8 0.0004 114 1

14-1 Cyfluthrin-1 1

14-2 Cyfluthrin-2 0.05–0.5 0.9982 2.8 1.8 1.1 0.0005 1

114.8
14-3 Cyfluthrin-3 1

14-4 Cyfluthrin-4 1

15-1 tau-Fluvalinate-1 1
0.05–0.5 0.999 2.8 1.4 0.7 0.0005 105
15-2 tau-Fluvalinate-2 1

16 β-BHC 0.05–0.5 0.9998 1 0.2 0.8 0.00007 89.2 2

17 γ-BHC 0.05–0.5 0.9999 1.4 0.3 0.9 0.00003 94.7 2

18 Pentachloronitrobenzene 0.05–0.5 0.9999 1.2 0.2 0.9 0.00003 91.6 2

19 Propanil 0.05–0.5 0.9999 4 1.1 1 0.0002 98.7 2

20 Vinclozolin 0.05–0.5 0.9999 1.9 1.3 0.8 0.00009 89.4 2

21-1 Endosulfan-1 2
0.05–0.5 0.9984 2.1 0.5 0.8 0.00008 94.8
21-2 Endosulfan-2 2

22 p,p’- DDD 0.05–0.5 0.9995 3.8 2 1 0.00006 96.4 2

23 Dicofol 0.05–0.5 0.9982 2 2.8 3.3 0.0006 95.6 2

24 Lambda-cyhalothrin 0.05–0.5 0.9991 2.1 0.4 0.9 0.0001 94.1 2

25 Permethrin 0.05–0.5 0.9987 2.3 1.3 2.2 0.0005 107.5 2

5
 Table 3. Data results for organochlorine pesticides analysis on an Agilent DB-5 column (continued).

% RSD (n = 8)
Linearity
Range 0.05 0.1 0.5 MDL %
No. Name (mg/kg) R2 mg/kg mg/kg mg/kg (mg/kg) Recovery Group
26-1 Flucythrinate-1 2
0.05–0.5 0.991 1.3 0.4 1 0.0005 92
26-2 Flucythrinate-2 2

27 Dicloran 0.05–0.5 0.9998 1.2 1.7 1.7 0.00006 80.1 3

28 Hexachlorobenzene 0.05–0.5 0.9997 0.9 1.8 0.5 0.00004 85.2 3

29 Chlorothalonil 0.05–0.5 0.9996 1 0.3 0.4 0.00006 82.7 3

30 Triadimefon 0.05–0.5 0.9997 0.9 1.1 0.6 0.00007 87.4 3

31 Procymidone 0.05–0.5 0.9995 0.6 0.5 0.6 0.0001 99.5 3

32 Butachlor 0.05–0.5 0.9997 0.7 0.5 0.5 0.0003 89.3 3

33 Dieldrin 0.05–0.5 0.9997 0.7 0.7 0.6 0.00004 85.7 3

34 Endrin 0.05–0.5 0.9996 0.8 0.7 0.5 0.00004 84.6 3

35 Chlorobenzilate 0.05–0.5 0.9983 2.3 1.6 0.3 0.0003 89.5 3

36 o,p’- DDT 0.05–0.5 0.9998 1.1 0.8 0.5 0.00007 94.1 3

37-1 Tetramethrin-1 3
0.05–0.5 0.997 2.2 1.1 2.4 0.0003 85.7
37-2 Tetramethrin-2 3

38 Fenpropathrin 0.05–0.5 0.9999 0.9 1.1 0.6 0.0002 90.72 3

39-1 Cypermethrin-1 3

39-2 Cypermethrin-2 1.9 0.8 1.1 0.0003 81.7 3

0.05–0.5 0.997
39-3 Cypermethrin-3 3

39-4 Cypermethrin-4 3

40-1 Fenvalerate-1 3
0.05–0.5 0.998 1.2 0.6 0.6 0.0003 93.9
40-2 Fenvalerate-2 3

41 Deltamethrin 0.05–0.5 0.9994 1.4 0.6 0.6 0.0002 86.6 3

6
Sample preparation QuEChERS sample preparation workflow

An apple sample was purchased from Weigh a 10 g sample (±0.01 g) into a 50 mL centrifuge tube.
a local grocery store. Ten grams of
homogenized apple sample were
weighed into a 50 mL centrifuge tube, Add spike solution, and vortex for one minute.*
and two ceramic homogenizers were
added to the sample. QC samples were
Add ACN (10 mL) and two ceramic homogenizers (p/n 5982-9313).
spiked with appropriate amounts of
spiking solution to yield QC samples
with a quantitative concentration of Add a QuEChERS extraction salt packet (p/n 5982-5650), and vortex for one minute
approximately 0.1 mg/kg. Ten milliliters
of acetonitrile were added to the tube. An
Agilent QuEChERS extraction salt packet Centrifuge for five minutes at 5,000 rpm at 10 °C.
(part number 5982‑5650) containing
4 g of MgSO4, 1 g of sodium chloride,
Transfer 6 mL of the upper layer to a dispersive tube (p/n 5982-5056).
1 g of Na-citrate, and 0.5 g of disodium
citrate sesquihydrate was added to
each centrifuge tube for extraction. An Vortex for one minute, and centrifuge for five minutes at 5,000 rpm at 10 °C.
Agilent QuEChERS general fruit and
vegetables dispersive SPE 15 mL tube
(part number 5982-5056) was used for Transfer 1 mL of the upper layer to a tube, then dry the final supernatant under nitrogen.
cleanup. For fruits and vegetables with
high pigments and fats, other types Use 1 mL of appropriate solvents** to reconstitute, vortex for one minute,
of QuEChERS packets are needed for and centrifuge for five minutes at 5,000 rpm at 10 °C.
extraction and cleanup. The details of the
sample preparation procedure are shown
in Figure 2. Transfer the extract to an autosampler vial.

Matrix blanks were prepared in the same * This is for the recovery test. For matrix blanks, skip this step.
** Using acetone for organophosphorus pesticides, while using hexane for organochlorine pesticides.
manner as the samples, except there
was no addition of spike solution. Figure 2. Flowchart of the QuEChERS extraction procedure for apple samples.

7
Results and discussion
Dimethoate Parathion-methyl
A Acetone
Matrix Ditalimfos
Disulfoton Chlorpyrifos
Organophosphorus Acephate Pirimiphos-ethyl
pesticides analysis Fenthion

The sample matrix has a great

influence on the results of pesticide Dicrotophos

analysis. Figure 3 shows a comparison Phoxim

of pesticide chromatograms in an
apple matrix blank and acetone. Blue
represents a standard prepared in
acetone, while red represents a standard
prepared in the apple matrix blank. It
B
shows that for some compounds, using Methamidophos Acetone
a matrix blank to dilute the working Matrix

solution can improve the sensitivity,

especially for some compounds that are
difficult to analyze, such as acephate and
methamidophos. Organophosphorus
pesticides, particularly polar pesticides
such as acephate and methamidophos,
tend to have broad peaks or tailing. The Figure 3. Comparison of chromatograms in apple matrix and acetone (approximately 0.1 mg/ kg) using an
peak shapes were also improved in the Agilent HP-50+ 30 m × 0.53 mm, 1 µm capillary GC column.
matrix blank.
Group 1
Simultaneous primary and confirmation 220 A HP-50+ 13
analysis from a single injection was 200
5
180 2
accomplished using a dual-FPD GC 160
6
4
system. An Agilent CFT 2-way splitter 140
11
78
without make-up device was used in 1
150 pA

120 9
this system. The 54 organophosphorus 100
3
12
10
pesticides were divided into four groups 80
60
for easy and accurate retention time 40
determination. Figures 4 to 7 illustrate 20
the analyses of groups 1, 2, 3, and 4 0
organophosphorus pesticide mixtures on 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
220 Time (min)
the HP-50+ and HP-1 columns. 200
B HP-1
13
180
160 2
140
150 pA

120
5 6
100
1 4 7 8 11 12
80 9
3
60
40 10
20
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
Time (min)

Figure 4. Chromatograms of group 1 organophosphorus pesticide standard solution (approximately

0.1 mg/kg) on a dual-column system using Agilent HP-50+ and HP-1 capillary GC columns.

8
Matrix-matched calibration standards 220 Group 2
A HP-50+
and spiked QC samples were prepared 200
14 27
by spiking appropriate standard solutions 180
160
into the matrix blank. For 44 compounds 19 26
140
analyzed, the linearity range was in the 15 20 22

150 pA
120 16 28
range of 0.05 to 0.5 mg/kg. For other 100 18 24
21
compounds such as phoxim, phosmet, 80
17
23
25

trichlorfon, pyrazophos, and so on, which 60

have low response factor, the linearity 40

20
range was 0.2 to 2 mg/kg. Table 2 shows
0
the details. Linearity across the range 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
studied gave R2 values of 0.992 for 220 Time (min)
B HP-1
all the organophosphorus pesticides. 200 14

Most of them have R2 values greater 180

160
than 0.999. Table 2 lists the correlation 27
140
coefficient for each of the pesticides on
150 pA

120 21
an HP‑50+ column. 100 19 26 28
15 24
20 22
Repeatability assessments at three 80 17 16
18
60 25
levels: low, middle, and high were 23
40
obtained for all compounds in apple 20
matrix. For most compounds, the low 0
level was 0.05 mg/kg, the middle level 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
Time (min)
was 0.1 mg/kg, and the high level was
0.5 mg/kg. For low response value Figure 5. Chromatograms of group 2 organophosphorus pesticide standard solution (about 0.1 mg/kg) on
compounds, the low, middle, and high a dual-column system using HP-50+ and HP-1 capillary GC columns.

levels were 0.2, 0.4, and 2 mg/kg except Group 3

30
naled and phosphamidon. Table 2 shows 260 A HP-50+
240
that the area RSDs were less than 5% for 220
all the compounds, which demonstrated 200
41
the accurate, precise, and stable 180
160
29
36 39 42
31
150 pA

performance of this system. 140 32 33

34 38
120 35
37
Signal-to-noise ratio (S/N) was used for 100
method detection limit (MDL) calculation. 80
40
60
A concentration of the lowest calibration 40
level was used to test the MDL, and the 20
0
values for all compounds are listed in 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
Table 2. Time (min)
260 B HP-1 39 34 36
33 35
240
220
200
180 29 30 42
160
150 pA

41
140 35
120 34
100 31 39 36
32 38
80 33 37
60 40
40
20
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
Time (min)

Figure 6. Chromatograms of group 3 organophosphorus pesticide standard solution (about 0.1 mg/kg) on
a dual-column system using HP-50+ and HP-1 capillary GC columns.

9
As described in the Sample preparation 170 A HP-50+ Group 4
section, QC samples were spiked with 160
150
appropriate amounts of spiking solution 140
130 49 54
to yield QC samples with quantitative 120 46-2
50
110
concentration of 0.1 mg/kg (for low 100 44 47
48

150 pA
response value compounds such as 90
80 51
43 45
phoxim, acquired at the 0.4 mg/kg 70
60
52 53
46-1
level). Recoveries were determined on 50
40
an HP-50+ column, and the results for 30
20
all organophosphorus pesticides were 10
between 70.4 and 118.2%. Table 2 0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
lists the recoveries for the individual Time (min)
pesticide. Most of the compounds, 200
B HP-1

even the polar compounds such as 180 48

49
52 47
50

acephate and methamidophos, have 160

54
51

good recovery data due to the excellent 140

extraction and cleanup process of 120 52

150 pA

47
QuEChERS, as shown in Figure 8.
100
49 50
80 44
46-1 46-2 48
60 53
45 51
43
40
20
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
Time (min)

Figure 7. Chromatograms of group 4 organophosphorus pesticide standard solution (approximately

0.1 mg/kg) on a dual-column system using Agilent HP-50+ and HP-1 capillary GC columns.

% Recovery for organophosphorus pesticides

140
120
100
% Recovery

80
60
40
20
0
Profenofos

Propetamphos
Paraoxon

Parathion
Isofenphos
Methidathion
Phosfolan-methyl
Dichlorvos
Acephate

Disulfoton
Dimethoate
Parathion-methyl
Chlorpyrifos
Pirimiphos-ethyl
Fenthion

Ditalimfos

Phosmet
Trichlorfon
Ethoprophos
Phorate
Omethoate
Diazinon
Fonofos
Chlorpyrifos-methyl

Fenitrothion
Bromophos
Bromophos-ethyl

Ethion

Coumaphos
Methamidophos

Famphur
Phosalone
Azinphos-ethyl
Naled
Sulfotep
Terbufos
Monocrotophos
Dichlofenthion
Fenchlorphos
Pirimiphos-methyl

Mevinphos

Phosphamidon

Azinphos-methyl
Trichloronate
Malathion

Quinalphos
Isocarbophos

Tetrachlorvinphos
Phosfolan
EPN
Phoxim

Triazophos

Pyrazophos
Dicrotophos

Figure 8. Recovery data of organophosphorus pesticides.

10
Organochlorine pesticides analysis Group 1
Similar to the analysis of 4,500 A DB-5 4 5
1
organophosphorus pesticides, 4,000

simultaneous primary and confirmation 3,500 6

13 14

analysis from a single injection was 3,000 7 8 11

accomplished using a dual-ECD GC 2,500 9

Hz
10
system for organochlorine pesticides 2,000

analysis. A CFT two-way splitter without 1,500

make-up device was used in this system. 1,000

2 12
11
15
13 14
Forty-one organochlorine pesticides were 500
3
divided into three groups. Figures 9 to 11 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34
illustrate the analysis of groups 1, 2, and Time (min)
3 organochlorine pesticide mixtures on 4,500
B DB-17
DB-5 and DB-17 columns. 4,000
3,500 54
1
3,000
6 8
2,500 7
Hz

9
2,000
10
1,500
1,000 13
3 12 15
500 14
2 11

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34
Time (min)

Figure 9. Chromatograms of group 1 organochlorine pesticide standard solution (0.1 mg/kg) on a

dual-column system using DB-5 and DB-17 capillary GC columns.

Group 2
5,500 17
A DB-5 18
17
5,000 18
4,500
4,000
16
3,500
3,000
Hz

2,500 22
2,000 21-2
16 20 24
1,500
1,000 19 21-1 26
500 23 25

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34
Time (min)
5,000 B DB-17
4,500 17
18
4,000
3,500
3,000
Hz

2,500
2,000 22
16
1,500 20 21 23,24
1,000 19 26
500 25

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34
Time (min)

Figure 10. Chromatograms of group 2 organochlorine pesticide standard solution (0.1 mg/kg) on a
dual‑column system using DB-5 and DB-17 capillary GC columns.

11
For the analysis of organochlorine 3,000 28
Group 3
38
A DB-5 33
pesticides, some compounds such 2,800 34
2,600 39
as cyfluthrin and cypermethrin have 2,400
isomers. The retention times of those 2,200
2,000
37
isomers were close, and it was difficult 1,800 27
29
35 37
1,600
to achieve baseline separation, as shown

Hz
1,400 30 36
in Figure 12. For these compounds, the 1,200
1,000
setting of integration parameters was 800
31 38 40
41
39
particularly important. Because the 600
400
32
37
standards purchased from the vendor 200
were also isomer mixtures, and no 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44
other compounds eluted between the 3,000
Time (min)
B DB-17 33
isomers, those isomers were integrated 2,800 34
2,600 28
as one peak for quantitative analysis. 2,400
36
Figure 13 shows that, in OpenLab CDS 2,200 3334
2,000
2.3 software, the Area Sum function can 1,800 27
35
help integrate the isomers. 1,600
Hz

29
1,400 36
1,200 30
1,000
800 31
600 38 39 40
32 35 41
400 37
200
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44
Time (min)

Figure 11. Chromatograms of group 3 organochlorine pesticide standard solution (0.1 mg/kg) on a
dual‑column system using Agilent DB-5 and DB-17 capillary GC columns.

2,000 Cyfluthrin
1,900
1,800
1,700
1,600
1,500
1,400
1,300
1,200
1,100
Hz

1,000
900
800
700
600
500
400
300
200

24.2 24.3 24.4 24.5 24.6 24.7 24.8 24.9 25.0 25.1 25.2 25.3
Time (min)

Figure 12. Chromatogram of cyfluthrin isomers using the Area Sum function for integration.

12
Matrix-matched calibration standards
and spiked QC samples were prepared
by spiking appropriate standard solutions
into the matrix blank. The spiking
concentration for calibration standards
were between 0.05 and 0.5 mg/kg in
apple matrix. The data were processed
with OpenLab CDS 2.3 software. Table 3
shows the results on a DB-5 column, the
R2 values >0.991 for all organochlorine
pesticides. The area RSD values for
eight replicates at three levels were
below 4%, with the typical RSD below
2%. Compared to the NY/T 761 method,
the optimized extraction and cleanup
procedure was validated by running
spiked samples at 0.1 mg/kg level.
Acceptable recoveries were achieved Figure 13. Integration table for cyfluthrin isomers.
for most of the analytes. Recoveries
were between 77.3 and 118.6%. Table 3
also shows the MDL results for the
41 compounds. S/N was used for MDL
calculation. The results were better than
the NY/T 761 method reference results.

% Recovery for organochlorine pesticides

140
120
100
% Recovery

80
60
40
20
0
p,p'-DDT

o,p'-DDT
Propanil
Vinclozolin
Bifenthrin
cis-Permethrin
Cyfluthrin
α-BHC
Simazine

δ-BHC
Heptachlor
Aldrin
o,p'-DDE
p,p'-DDE
o,p'-DDD

Iprodione

tau-Fluvalinate
β-BHC
γ-BHC
Pentachloronitrobenzene

Endosulfan
p,p'-DDD
Dicofol
Lambda-cyhalothrin
Permethrin
Flucythrinate
Dicloran
Hexachlorobenzene
Atrazine

Triadimefon
Procymidone
Butachlor
Dieldrin
Endrin

Tetramethrin
Fenpropathrin
Cypermethrin
Fenvalerate
Deltamethrin
Chlorothalonil

Chlorobenzilate

Figure 14. Recovery data of organochlorine pesticides.

13
Conclusion
An Agilent 8890 GC configured with four
detectors (two FPDs and two ECDs)
was used to screen organophosphorus
and organochlorine pesticides in fruit
and vegetables. Splitting the samples
into two different columns then to
two detectors facilitated selectivity,
identification, and confirmation of
organophosphorus pesticides and
organochlorine pesticides from single
injections of each extract, increasing
laboratory productivity.
This Application Note demonstrates
excellent sensitivity, area repeatability,
peak shape, and resolution for both
organophosphorus and organochlorine
pesticides, which shows that this
four‑detector system is an ideal platform
for the NY/T 761-2008 method.
www.agilent.com/chem
This information is subject to change without notice.
© Agilent Technologies, Inc. 2019
Printed in the USA, October 8, 2019
5994-1215EN
References
1. China National Standard NY/T
761-2008, Determination of
Organophosphorus, Organochlorine,
Pyrethroid and Carbamate Residues
in Vegetables and Fruits.
2. China National Standard GB/T
19648-2006, Determination of
500 Pesticides and Metabolites
Residues in Vegetables and Fruits,
Gas Chromatography/Mass
Spectrometry Method.
3. China National Food Safety Standard
GB 23200.113-2018, Determination
of 208 Pesticides and Metabolites
Residues in Foods of Plant Origin,
Gas Chromatography-Tandem Mass
Spectrometry Method.