(EXPERIMENT 7)

ANALYSIS OF CHLORPYRIFOS IN WATER BY

SOLID-PHASE EXTRACTION (SPE) AND GAS
CHROMATOGRAPHY-ELECTRON CAPTURE
DETECTOR (GC-ECD)
NAME:

MOHAMAD NOR AMIRUL AZHAR BIN KAMIS

STUDENT ID:

2014647344

PARTNERS NAMES: 1.

MOHAMAD HAMIZAN BIN MOHD ISA

2.

MOHAMAD SHAFIQ BIN PARMAN

3.

MOHAMAD AZMIZAM BIN MOHAMAD

NOOR

DATE OF EXPERIMENT:

17/10/2014

DATE OF SUBMISSION:

8/12/2014

INTRODUCTION:

Figure 1: chlorpyrifos
Chlorpyrifos is a crystalline organophosphate insecticide that acts on the nervous
system of insects by inhibiting acetylcholinesterase. Chlorpyrifos is moderately toxic
to humans and the exposure will cause neurological effects such as persistent
developmental disorders autoimmune disorders and children mental development
retard due to the exposure during pregnancy. In agriculture, chlorpyrifos remains as
one of the most widely used as the pesticide. Solid-phase extraction (SPE) is an
extraction method that uses solid and liquid phase to isolate analyte from solution. It
is a rapid and selective sample preparation. The purpose of using the SPE is to
purify, trace enrichment, solvent exchange and derivatization. The advantages of the
SPE are high recovery of analyte, concentration of analyte, ease of automation and
reduction of organic solvent consumption. The general procedure of SPE is the
loading of solution through SPE phase, wash away undesired components, and
lastly wash off the desired analytes with other solvent into the collection tube. Steps
of SPE are from column solvation and equilibration, sample loading, interference
elution and lastly, the analyte elution. The objectives of this experiment are to
calculate the amount of chlorpyrifos and each waste water sample and also the
percentage of recovery.

EXPERIMENTAL:
a. Solid-phase extraction procedure:
1. 10mL of methanol was let to be passed the C18 SPE cartridge to activate
the column.
2. 6mL of deionized water was let to be passed the cartridge without applying
the vacuum to remove the excess solvent prior to adding the sample.
3. The filtered water sample (50mL) was let to be passed through the
preconditioned column using a vacuum manifold at about 48-55 drops/min.
The column was not let dry during the sample enrichment step.

4. The column was dried by using the vacuum for 15 minutes.

5. The interference was removed by eluting the column with 10mL of
deionized water and the column again was dry by using vacuum for 30
minutes.
6. The pesticide was eluted by using 5mL of hexane and then was
concentrated into about 1mL by gently blowing with nitrogen.
b. Instrument set-up:
Injector temperature: 280C
Detector temperature: 300C
Carrier gas flow rate: 20.0mL/min (nitrogen)
Column temperature: initial temperature 165C for 3 minutes, increase to
260C at 3C/min with a final time of 2 minutes.
c. Quantitative analysis of chlorpyrifos:
1. 1L of sample was injected onto the column and to get the reproducible
peak, the injection was repeated for each sample.
2. 1L of standard chlorpyrifos (30ppm) was injected and the injection was
repeated to get reproducible peak area.
3. By using the data from the standard solution of chlorpyrifos, the
concentration of chlorpyrifos in the samples were calculated.

RESULT AND DISCUSSION:

A. Comparison in retention time of standard and samples:
Retention
Sample
Retention time of sample
time of
(min)
standard
(ppm)

6.901

Average
retention
time of
sample
(min)

Trial 1
6.919

Trial 2
6.919

6.919

2
3

6.917
6.920

6.917
6.920

6.917
6.920

B. Calculation of response factor for standard compound:

Response Factor (RF) =
=
= 4.975 10-5ppm/Hz*s

C. Amount of chlorpyrifos in samples:

Amount of chlorpyrifos in sample = RF standard peak area sample
Sample

1
2
3

Area (Hz*s)

Trial 1
270559
247838
302817

Trial 2
272510
245646
306761

Average
area (Hz*s)

Amount of
chlorpyrifos
(ppm)

270534.5
246742
304789

13.46
12.28
15.16

Average amount of chlorphyrifos in sample =

= 13.63ppm
D. Percentage of recovery of samples:
Percentage recovery =
Amount of chlorpyrifos in standard = 30ppm
sample
Amount of chlorpyrifos
in sample (ppm)
1
13.46
2
12.28
3
15.16

100%
Percentage recovery
(%)
44.87
40.93
50.53

Average percentage recovery of sample =

= 45.44%
In this analysis, GC with Electron Capture detector is used because the analyte to be
analysed is halogenated compound. ECD only can detect analytes which contain
electronegative functional groups that can capture electrons such as halogens,
peroxides, quinones and nitro groups. The disadvantage of ECD is it involve
radioactive component. The amount of chlorpyrifos in samples is calculated by using
response factor calculation that base on the standard compound. The amount of
chlorpyrifos in each sample does not show big different, it show that same procedure
was carried out to each sample. The average amount of chlorpyrifos in the samples
is 13.63ppm, that is about half from the amount of chlorpyrifos in the standard. The
percentage recovery calculated is 45.44% in average. In order to get higher amount
of chlorpyrifos or the percentage recovery, the SPE must be carried out carefully so
it will extract more chlorpyrifos efficiently.

CONCLUSION:
The average amount of chlorpyrifos in sample is 13.63ppm and the percentage
recovery is 45.44%.

REFERENCES:
1. Norashikin S., Ruziyati T., Mardiana S. (2012), Analytical Separation
Methods Laboratory Guide (2nd edition), 3/10/2014.
2. Clorpyrifos, 30/10/2014, http://en.wikipedia.org/wiki/Chlorpyrifos.
3. Mardiana Saaid, 30/10/2014, Electron-Capture Detector (ECD) lecture notes.