1.

0 INTRODUCTION

High levels of pesticides are used every year in the production and post-production
treatments of agricultural commodities. Chlorpyrifos (CPF) is one of the most
frequently used organophosphorus pesticides (OPPs) in agriculture and in households.
OPPs are mostly sprayed over crops or applied to soils, leading to their direct transfer
from drainage of agricultural lands to other parts of surrounding environments,
including ground and surface waters. Therefore, there is an increasing concern
regarding the determination of these compound.

Chlorpyrifos is moderately toxic and classified in Toxicity Category II for all exposure
routes. According to the United States Environmental Protection Agency, exposure to
CPF could result in neurotoxicity in animals and humans, decreased birth weight of
babies and increased risk of lung cancer. The maximum permissible level for CPF in
fresh water is 0.041 µg/L, according to the USEPA water quality criteria.

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 in each
waste water sample and the percentage of recovery.
2.0 METHODOLOGY

Condition C18 SPE cartridge by passing 10 ml of methanol. The cartridge was

rinsed by passing 6 ml of deionized water without applying vacuum. The filtered water
sample (50 ml) were passed through the preconditioned column using a vacuum
manifold at 6 ml per min (48 drops/min). 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 vacuum dried again for 10 minutes. The pesticide was eluted
using 5 ml of hexane. Concentrate to 1 ml by blowing down using gentle nitrogen and
the sample ready for GC analysis.

The instrument was set-upped, temperature’s injector was 280°C, detector

temperature was 300°C, flow rate of carrier gas was 20.0 ml/min (nitrogen) and
column temperature was 165°C for 3 minutes initial temperature, then increased to
260°C at 3°C/min with a final time for 2 minutes.

For the quantitative analysis of chlorpyrifos, 1µL of sample was injected

onto the column. The injection was repeated to get reproducible peak areas. 1 µL of
standard chlorpyrifos was injected. The injection was repeated to get reproducible
peak area. The concentration of chlorpyrifos in the sample was calculated by using
the data from the standard solution.
3.0 RESULT AND DISCUSSION

A. Comparison in retention time of standard and samples

Retention Retention time of sample Average
time of (min) retention time
Sample
standard of sample
Injection 1 Injection 2
(min) (min)
1 6.724 6.725 6.7245
6.711 2 6.717 6.721 6.719
3 6.720 6.721 6.7205

B. Calculation of response factor for standard compound

𝑎𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝑐ℎ𝑙𝑜𝑝𝑦𝑟𝑖𝑓𝑜𝑠
𝑟𝑒𝑠𝑝𝑜𝑛𝑠𝑒 𝑓𝑎𝑐𝑡𝑜𝑟, (𝑅𝐹) =
𝑝𝑒𝑎𝑘 𝑎𝑟𝑒𝑎
30 𝑝𝑝𝑚
=
1.11956 𝑋 106 𝐻𝑧𝑠
= 2.6796 𝑋 10−5 𝑝𝑝𝑚/𝐻𝑧𝑠

C. Amount of chlorpyrifos in sample

Area (Hz*s) Amount of
Average area
Sample chlorpyrifos
Injection 1 Injection 2 (Hz*s)
(ppm)
1 537482 546153 541817.5 14.52
2 110798 113080 111939.0 3.00
3 241467 244626 243046.5 6.51

Average amount of chlorpyrifos

14.52+3.00+6.51
= 3

= 8.01 ppm
D. Percentage of recovery sample
amount of chlopyrifos in sample
% recovery = amount of chlophyrifos in standard x 100

Amount of chlopyrifos in standard = 300 ppm

Amount of chlorpyrifos
Sample Percentage recovery (%)
in sample (ppm)
1 14.52 48.40
2 3.00 10.00
3 6.51 21.70

Average % recovery
48.40+10.00+21.70
= 3

= 26.70 %
4.0 DISCUSSION

In this experiment, the chlorpyrifos was analysed in water by using

solid-phase extraction (SPE) and gas chromatography-electron capture detector (GC-
ECD). Based on the result, each sample was injected for two times. For sample 1 the
retention time that has a nice peak is at 6.724 minutes and 6.725 minutes. For the sample
2, the nice peak was observed at retention time 6.717 minutes and 6.721 minutes. Lastly,
the sample 3 has nice peak at retention time 6.720 minutes and 6.721 minutes for each
injection respectively. The retention time become longer from sample 1 to sample 3.

The response factor was calculated for this sample. The response factor is
2.6796 𝑋 10−5 𝑝𝑝𝑚/𝐻𝑧𝑠. This response factor was calculated with formula the sample
amount over the peak area for the standard solution. The sample amount for the standard
that was used is 30 ppm. Then, by using the value of response factor, the amount of
chlorpyrifos in the sample can be calculated by multiplied the response factor that
calculated before with the peak area of each sample, sample 1, sample 2 and sample 3.
The amount of chlorpyrifos in the sample 1 is 14.52 ppm. While in the sample 2 the
amount is 3.00 ppm and last in sample 3 is 6.51 ppm.

Then, the percentage recovery also was calculated by divide the amount of
chlorpyrifos in sample with the amount of chlorpyrifos in standard then multiplied it
with 100. The percentage recovery for sample 1 is 48.40 %, sample 2 is 10.00 % and
lastly sample 3 is 21.70 %.
5.0 CONCLUSION

Based on the experiment, the amount of chlorpyrifos in each waste water sample are
14.52 ppm for sample 1, 3.00 ppm for sample two and 6.51 ppm for sample 3 while the
percentage of recovery for sample 1,2 and 3 are 48.40 %, 10.00 % and 21.70 %
respectively.
6.0 REFERENCES

Nor’ashikin S., Ruziyati T., Mardiana S. (2012), Analytical Separation Methods

Laboratory Guide (2nd edition)

Studia Universitatis Babes-Bolyai Chemia. (2014), Determination of chlorpyrifos

in surface water using SPE-DI-SPME/GC-ECD. Retrieved from
https://www.researchgate.net/publication/287299453_Determination_of_chlorpyrifo
s_in_surface_water_using_SPE-DI-SPMEGC-ECD

Azhar. A., (2014), Analysis Of Chlorpyrifos In Water By Solid-Phase Extraction

(SPE) And Gas Chromatography-Electron Capture Detector (GC-ECD). Retrieved
from https://www.scribd.com/doc/249342246/ANALYSIS-OF-CHLORPYRIFOS-
IN-WATER-BY-SOLID-PHASE-EXTRACTION-SPE-AND-GAS-
CHROMATOGRAPHY-ELECTRON-CAPTURE-DETECTOR-GC-ECD