Analysis of polyaromatic hydrocarbons in water by headspace solid phase micro-extraction coupled with

capillary gas chromatography

Brannigan du Preez, 18387918

CHM 314

Stellenbosch University

Introduction

Headspace solid phase micro-extraction is an efficient procedure in preparing samples in laboratories or on site
operations, where the analyte is required to be investigated. This process involves the dispersion the extracted
phase onto a solid support which is exposed to the sample for a certain period of time. Equilibrium is reached
between the solid and the extracting phase, whereas in pre-equilibrium extraction the amount of analyte
extracted from the sample can be related to time. Quantification of the extracted analyte can be performed based
on the time it takes for the analyte to accumulate onto the solid fused silica coating.

The above-mentioned procedure can be coupled to capillary gas chromatography, whereby a liquid sample is
injected into an oven, through which it vapourised to passed through a column (moving as the mobile phase by
means of a carrier gas), through which the individual compounds within the sample can be separated according
to their respective affinities to the particular stationary phase used. A chromatographic plot is constructed by the
detector, usually performed by a computer, in order to indicate the retention times of each compound. The
higher the retention time on the chromatogram, the lower the affinity the compound has for the stationary phase
used.

Thus, coupling of these techniques enable the analyses of microsamples of low concentrations, through the
micro-extraction of analytes from a gaseous or liquid phase using HS-SPME; consequently, as previously
mentioned, CG-chromatography allows the separation of mixtures according to their affinities to a particular
stationary phase, which in this experiment, are poly-aromatic hydrocarbons within water. It is useful to be aware
of the potential environmental and carcinogenic effects these compounds possess. This experiment will enable
the presence of such substances, within water, to be determined. These compounds include naphthalene, fluorine
and phenanthrene using 1.4-dimethylnaphthalene as an internal standard.

HS-SPME is a reasonably good method by which micro-extraction is performed, as it uses less solvent and it
can be used to detect compounds with concentrations as small as g/L.

The objectives in this experiment include the extraction of analytes from calibration samples (and unknown
samples using HS-SPME) and the subsequent separation and identification of these samples using CG-analysis.

A calibration curve is drawn using the peak area of the compounds at their respective concentrations, upon
which the unknown poly-aromatic hydrocarbons are quantified within the sample.

Sample preparation

SPME/GC

The sample preparation for SPME is separated into two stages upon which the first, sorption, involved piercing
the sample septum with the injector, upon which the fiber extract containing the analyte was exposed to the
headspace (by which vaporisation onto the coating could occur) upon which the fiber was retracted. The second
procedure, desorption, involved the piercing of the GC septum, through the GC inlet, by which the fiber was
exposed to release the analyte, into the vaporizing inlet of the -------- after which the fiber was retracted.
ProcedureT

A water sample containing an unknown amount of poly-aromatic hydrocarbons were assigned to be identified,
whereas an internal standard of 1.4-dimethylnaphthalene was used. Target analytes were extracted according to
SPME within the calibration and the unknown sample, followed by capillary GC-FID analysis. A calibration
curve was obtained for the standards, which were used to quantify the sample.

HS-SPME: SPME was performed using an SPME device, which was equipped with a 100m PDMS fiber,
which was conditioned for hours at 250C within the splitless injector.

The extractions were performed within a 20 ml headspace vial which was kept at room temperature. The sample
was extracted for 30 minutes, whereby the sample was stirred at 1000 rpm, using a magnetic stirrer and a
Teflon-coated stirrer bar.

CGC-FID: GC-FID was performed using an Agilent 5890 GC, containing a split/splitless injector as well as a
detector. For the separation process, a 30 m capillary column (with a film thickness and internal diameter of
0.25mm and 0.250 mm, respectively. Temperature programming proceeded using 60C for one minute,
increasing it to 250C (at 15C per minute) for five minutes. The injector time was set to 280C at a two-minute
splitless time. The carrier gas, whose pressure and flow rate were 100 kPa and 1 ml/minute respectively, used
was helium. The FID temperature was also 280C, with gas pressures of 100 kPa of hydrogen and 300 kPa of air
containing flow rates of 35 and 350 ml/minute, respectively.

Results

Concentration of
Peak 1 Peak 2 Peak 3 Peak 4
Standards (ppb)
10 6594,5 176632,2 12572,5 4425,5
30 16973,4 148531,8 26624,9 12096,9
50 34338,6 161142,1 41942,8 23351,5
75 64952,3 186867,7 79716,9 45009,1
150 108968,1 154349,0 136427,0 81669,1
Unknown sample 1 18344,6 65709,4 38178,9 58505,8
Unknown sample 2 10700,7 43655,9 24933,8 40600,3
Unknown sample 3 18284,7 68816,8 38895,0 61963,2

Table 1 represents the various peak areas of the compounds at different concentrations (in ppb). Note that peak
2 was used as an internal standard.

Corrected Peak Area

 Conc. of Standards
Peak 1 Peak 3 Peak 4
(ppb)
10 0,0373 0,0712 0,0251

30 0,1143 0,1793 0,0814

50 0,2131 0,2603 0,1449

75 0,3476 0,4266 0,2409

150 0,7060 0,8839 0,5291

Unknown sample 1 0,2792 0,5810 0,8904

Unknown sample 2 0,2451 0,5711 0,9300

Unknown sample 3 0,2657 0,5652 0,9004

Table 2 represents the corrected peak areas of the compounds at their various concentrations

Figure 2 shows the