EXPERIMENT 5:

ANALYSIS OF CHLORPYRIFOS IN WATER BY SOLID-PHASE

EXTRACTION (SPE) AND GAS CHROMATOGRAPHY- ELECTRON
CAPTURE DETECTOR
(GC-ECD)

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 auto-
immune 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.

PROCEDURE
a) Filter Water Sample Through A Glass Fibre Filter Paper
It was prepared well by Supervisor.

b) Solid-phase Extraction Procedure

C18 SPE cartridge was conditioned by passing 10 mL of methanol. The cartridge was rinsed by
passing 6 mL of deionized water without applying vacuum. Then, the filtered water sample
was passed through the preconditioned column using a vacuum manifold at ~6 mL per min
(about 48 drops min-1). The column should not be allowed to dry during this sample enrichment
step. The column was dried by vacuum for 15 minutes. The interference was removed by
eluting the column with 10 mL of deionized water and the cartridge was dried by vacuum for
30 minutes. The cartridge was placed into tube rack. After that, the pesticide was eluted using
5 mL of hexane. The sample was concentrated to 1 mL by blowing down using gentle nitrogen
and the sample were sealed by using parafilm for GC analysis.

c) Instrument Set-up
Injector Temperature: 280oC
Detector Temperature: 300oC
Carrier Gas Flow Rate: 20.0 mL min-1 (Nitrogen)
Column Temperature: Initial Temperature 165oC for 3 minutes, increase to 260oC at 3oC min-
1 with final time of 2 minutes

d) Quantitative Analysis Of Chlorpyrifos

1 µL of sample was injected onto the column. The injection was repeated to get reproducible
peak areas. Then, 1 µL of standard chlorpyrifos were injected and repeated to get reproducible
peak area. The concentrations of chlorpyrifos in the sample were calculated using the data from
standard solution.

RESULTS

A. Comparison in retention time of standard and sample;

Retention time of Sample Retention time of sample (min) Average retention

standard (ppm) Trial 1 Trial 2 time of sample
(min)
 5.145 1 5.148 5.148 5.148
2 5.149 5.150 5.150
3 5.146 5.146 5.146

A. Calculation of response factor for standard compound;

= 30 ppm/ 247.1895 Hz*s

= 0.0333 ppm/Hz*s

A. Amount of Chlorpyrifos in samples;

Amount of chlorpyrifos in sample= Peak area of sample x Response Factor (standard)

Sample Area (Hz*s) Average area Amount of

Trial 1 Trial 2 (Hz*s) chlorpyrifos(ppm)
1 443.1896 459.2441 451.2169 15.0255
2 421.0789 404.0869 412.5829 13.7390
3 181.9961 236.5716 209.2839 6.9692

Average amount of chlorpyrifos in sample (ppm):

=(15.0255+13.7390+6.9692)/3 = 11.9112 ppm

A. Percentage of recovery of samples;

DISCUSSION

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 11.9112 ppm, that is about half from the amount of chlorpyrifos in the
standard. The percentage recovery calculated is 39.71% 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 11.9112 ppm and the percentage recovery
is 39.71%.
REFERENCE

1. Nor’ashikin S., Ruziyati T., Mardiana S. (2012), Analytical Separation Methods Laboratory
Guide (2nd edition).
2. https://earthjustice.org/features/what-you-need-to-know-
about_chlorpyrifoshttps://en.wikipedia.org/wiki/Chlorpyrifos

http://fareastzareul1994.blogspot.com/2018/05/experiment-5-analysis-of-chlorpyrifos.html