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Lehrstuhl für Siedlungswasserwirtschaft

Ingenieurfakultät Bau Geo Umwelt


Technische Universität München

Gas Chromatography

PD Dr. J. Graßmann; PD Dr. T. Letzel


Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Gas chromatography allows the separation of volatile compounds by means of a mobile and a
stationary phase. The component of an injected sample are vapourised and transported onto the
column by a flow of inert gas (mobile phase), e.g. helium, argon, nitrogen, carbon dioxide.
Generally two types of columns are available, which either contain a solid (gas-solid
chromatography, GSC) or a liquid stationary phase (gas-liquid chromatography, GLC). Liquid
stationary phases furthermore divides into two general types. The first one containes a finely and
evenly divided inert support material („packed column“), which is coated with the liquid stationary
phase. In contrast „capillary columns“ are tubes with the stationary phase adsorbed to the inert
inner wall of the column.

Capillary column Packed (open tubular)


column

10 to 100m length and Typically 1 to 10m


0.1 to 0.75mm inner length and 2 to 4mm
diameter inner diameter

Liquid stationary
phase
PD Dr. J. Graßmann; PD Dr. T. Letzel
Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
The separation is based on the analytes solubility in the liquid stationary phase (GLC) (or their
adsorbance on the solid stationary phase (GSC)). An analyte, which does not dissolve in the
stationary phase, would pass through the column unretained at the speed of the inert gas (carrier
gas) (= early elution). Analytes, which are likely to dissolve in the stationary phase have less
contact with the passing gas flow and would therefore need more time to pass the column and to
reach the detector (= late elution).

„Compounds are separated according to their partition coefficient“

Carrier gas
with analytes
Good solubility of analyte Poor solubility of analyte
in the stationary phase in the stationary phase
 Slowly moving  rapidly moving
PD Dr. J. Graßmann; PD Dr. T. Letzel
Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Parts of a Gas Chromatograph

Injector

Flow control Detector

Column Data recording &


processing

Carrier gas Oven

PD Dr. J. Graßmann; PD Dr. T. Letzel


Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Injection port temperature is kept at constant
temperature 20 to 50°C above the analytes
boiling point to vaporize the sample.

Either liquid or already gaseous samples can


the introduced to the column.

Injection modes:
1) Split injections
The volume of the vaporized sample is split
before entering the column (small volume enters
the column)
2) Splitless injection (split outlet is closed)
The entirety of the vaporized sample is allowed to
enter the column. Therefore the sample needs to
be properly diluted to avoid column overloading.
3) On column injection
At a temperature approx. 20°C below the analytes
boiling point the sample is directly injected onto
the column, whereupon the sample is vaporized
by increasing the temperature.
PD Dr. J. Graßmann; PD Dr. T. Letzel
Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Headspace gas chromatography

Headspace technique is used for the analysis of volatile and semi-volatile analytes. A sealed vial
containing the sample is heated moderately (e.g up to 60°C), which causes highly volatile
analytes to enter the gas phase (= headspace volume). The more volatile a compound, the higher
its concentration in the headspace. The less volatile compounds will remain in the liquid sample.
The headspace vapor can now be extracted to be injected onto the gas chromatographic column.
The pre-separation results in less non-volatile compounds entering the GC, which results in a
much cleaner chromatography.

Headspace Volatile compounds


migrate into the head-
space phase
heating
Sample Non-volatile compounds
remain in liquid phase

PD Dr. J. Graßmann; PD Dr. T. Letzel


Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Different types of capillary columns (1)
Packed columns Packed with stationary phase coated diatomaceous earth particles.
Higher capacity than most capillary columns, but lower efficiency,
broader peaks and inferior peak separation compared to capillary
columns.
PLOT Thin layer of porous particles adhered to the inner wall of the
(Gas solid column. Adsorption and desorption as well as size exclusion effects
chromatographie) result in separation of the analytes.
WCOT Most widely used GC columns. The inner wall of the tube is coated
with the stationary phase.
SCOT Inner wall of the capillary is coated with a thin layer of support
material (e.g. diatomaceous earth) onto which the stationary phase
is adsorbed
FSOT Polyimide coating improves stability compared to WCOT.

PD Dr. J. Graßmann; PD Dr. T. Letzel


Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Different types of capillary columns (1)
Fused silica tube
Porous-layer open
tubular column Polyimide coating
(PLOT)

Stationary
phase

Support-coated open
Wall-coated open tubular column
tubular column (SCOT)
(WCOT)
Fused solica open
tubular column
(FSOT)
Porous solid support Liquid stationary
phase
PD Dr. J. Graßmann; PD Dr. T. Letzel
Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Examples for stationary phases
50%-Cyanopropylphenyl)-dimethylpolysiloxane
Dimethylpolysiloxane  polar
 Non-polar
CH3 CH3

O O Si O Si
Si
CH3 (CH2)3
CH3
50% 50%
100% CN
(35%-Phenyl)-methylpolysiloxane
 semi-polar In general stationary phase ought to
be chemically inert, non-volatile,
CH3 stable even at high temperatures and
chemically similar to the compounds
O Si O Si to be analyzed, e.g. non-polar
compounds and non-polar stationary
CH3 phase
75% 35%
PD Dr. J. Graßmann; PD Dr. T. Letzel
Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Derivatization

Non-volatile compounds can be chemically converted to a volatile form. For instance


amino acids are not suitable for GC. They can be derivatized to decrease their polarity
and increase their volatility. Polar groups like hydroxyl groups (=OH), amino groups
(NH2) or thiol groups (SH) can be replaced by non-polar moieties, e.g. through
sinylation with trimethylsilyl derivatives.

Drawing of chemical structures was performed with „MarvinSketch 14.8.25.0“, ChemAxon (www.chemaxon.com)

PD Dr. J. Graßmann; PD Dr. T. Letzel


Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Detector (1)
Flame ionization detector (FID)
FID is a destructive detector. The separated
compounds from the column are combusted at
Collector high temperatures by a hydrogen flame. The
hereby produced positively charged ions are
- detected by a negatively charged collector
Power
supply electrode. In contrast, the released electrons flow
Flame
+ towards the positively charged burner head.
Igniter The generated current is then measured.
Most carbon atoms, except for carbonyl and
Air
carboxyl groups, generate a signal, which is
H2
independent of the compounds structure. FID
detector is therefore a universal detector for
column organic compounds.

PD Dr. J. Graßmann; PD Dr. T. Letzel


Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Detector (2)
Nitrogen phosphorus Highly sensitive detector, which detect the presence of compounds containing
detector (NPD) nitrogen or phosphorus. The ionization of the analyte is achieved using thermal
energy.
Thermal conductivity (TCD) Comparable insensitive detector. Changes of the thermal conductivity due to the
presence of eluting compounds in the carrier gas flow are detected in comparison
to a reference flow of carrier gas.
Electron capture detector Very sensitive detector, which is widely used for the analysis of halogenated
(ECD) compounds (e.g. Pesticides). Detection is based on the generation of negatively
charged analytes, which are able to capture electrons. This includes mainly
compounds with halogen atoms, which have high affinity to electrons. For other
compounds derivatization might be necessary to be able to detect them.
Flame photometric detector Sensitive detection of volatile phosphorus and sulphur compounds. The detectro
(FPD) captures the chemiluminescent emission from excited sulphur and phosphorus
species at a specific wavelength.
Photo ionization detector Sensitive and nondestructive detection. Ionization of compounds by means of UV
(PID) light. The separated analytes absorb the light and ionize. Electrodes then detect
the charged compounds and the current is recorded. This technique is often
selective and does not capture all of the contained compounds in a sample.
Mass spectrometric Detection of a wide range of ionizable compounds via their mass-to-charge ratio
detection (compare slides „Mass spectrometry“).
PD Dr. J. Graßmann; PD Dr. T. Letzel
Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
2-Dimensional Gas Chromatography GCxGC
Two different GC columns (often polar and nonpolar) are connected in series. The compounds are therefore
retained based on two different mechanisms . The analytes are separated by the first column and detected,
whereupon the efluent is trapped, focused and injected onto the second column (= heart-cut). Repeated
trapping and injection leads to a 2-dimensional chromotogram.

1st dimension data recording &


processing
Injector
2nd dimension data
recording &
processing

Cut

Column 1 Column 2

PD Dr. J. Graßmann; PD Dr. T. Letzel


Lehrstuhl für Siedlungswasserwirtschaft
Ingenieurfakultät Bau Geo Umwelt
Technische Universität München

Gas Chromatography
Things to consider....
Volatility of compound GC only suitable for analysis of volatile compounds

Temperature stability of compounds Stability might be improved by derivatization of the compounds

Molecular weight of analytes < 1000 Da

Stable temperature (=isotherm) or Considering the analytes boiling points and the temperature stability of the
temperature programm analytes as well as the column. High temperatures may cause column
bleeding, due to the degradation of the stationary phase = high background
signals.
Selection of suitable column and Crucial, since carrier gas acts only as transport medium (in contrast to LC)
stationary phase and gas flow and does not contribute to the chromatographic separation
Injection and injected sample Choose between splitless, split or column injection. Sample dilution might
volume be necessary to avoid column overloading.
Chemical properties of the analytes e.g. presence of polar groups; functional groups

Number of analytes to be separated Similar or highly different boiling points affect the temperature or the
temperature program applied
Efficiency of column WCOT (capillary) > SCOT (capillary) > packed columns

Column length The longer the column, the longer the analysis time, but the better the
resolution. The more complex the sample, the longer the required column.
PD Dr. J. Graßmann; PD Dr. T. Letzel