Gas Chromatography

Gas chromatography is a technique used for

separation of volatile substances, or substances
that can be made volatile, from one another in a
gaseous mixture at high temperatures. A sample
containing the materials to be separated is injected
into the gas chromatograph. A mobile phase
(carrier gas) moves through a column that
contains a wall coated or granular solid coated
stationary phase. As the carrier gas flows through
the column, the components of the sample come
in contact with the stationary phase. The different
components of the sample have different affinities
for the stationary phase, which results in
differential migration of solutes, thus leading to
separation 2
Advantages of Gas Chromatography
• Requires only very small samples with little
preparation
• Good at separating complex mixtures into
components
• Results are rapidly obtained (1 to 100 minutes)
• Very high precision
• Only instrument with the sensitivity to detect volatile
organic mixtures of low concentrations
• Equipment is not very complex (sophisticated oven)
 Gas Chromatography
In Gas Chromatography (GC), gaseous analyte is transported
through the column by a gaseous mobile phase, called the carrier
gas. The stationary phase usually is a non-volatile liquid or a solid
and the analytes are gases or volatile liquids.

GC
GLC GSC
Partition Adsorption
Stationary phase is a non-
The analyte is adsorbed
volatile liquid coated on the
directly on solid particles of
inside of the column or on a
fine solid support. stationary phase.
 Carrier gas
Block Diagram
flow
sample
meter
oven

data
carrier system
injector column detector
gas /readout
/display

reference gas

Carrier gas – main purpose of the gas in GC is to

move the solutes along the column, mobile
phase is often referred to as carrier gas.
Common carrier gas: include He, Ar, H2, N2
 Carrier Gases
• H2, He give better resolution (smaller plate height) than N2 at
high flow rates because solutes diffuse more rapidly through
H2 and He than through N2, so the Cu term is reduced.
• He is the most common gas because it is compatible with all
detectors.
• H2 ‘s drawback is that it can catalytically react with
unsaturated compounds on metal surfaces. It also forms
explosive mixtures with air when H2 > 4% vol.
• Impurities in the carrier gas degrade St. phase: High quality
gases should be used & they should be passed through
purifiers to remove O2 and H2O & traces of organic
compounds prior to entering the column. Steel or Copper
tubing rather than plastic or rubber should be used because
they do not release products in the gas stream.
 Injector
flow
sample
meter
oven

data
carrier system
injector column detector
gas /readout
/display

reference gas

• A GC syringe penetrates a septum to inject sample

into the vaporization camber
• Instant vaporization of the sample, 280 C
• Carrier gas transports the sample into the head of
the column
 Split (100:1) vs. Splitless (100:90)
• Usually operated in split mode unless sample limited
• Chromatographic resolution depends upon the width of the sample
plug
• In splitless mode the purge valve is close for 30-60 s, which means
the sample plug is 30-60 seconds
• As we will see, refocusing to a more narrow sample plug is possible
with temperature programming

Purge valve Purge valve

He open He
closed

GC column GC column
 Effect of operating parameters in split and splitless injections

Split and splitless injections of a solution containing 1 % vol methyl

isobutyl ketone and 1 % vol. p-xylene in dichloromethane.

© Dr. Rasha Hanafi, GUC Lecture 9 – Chromatography, 13-11-2012 10

 Head space analysis
Headspace' is the gas space above the
sample in a chromatography vial.

Volatile sample components diffuse into

the gas phase, forming the headspace gas.

Headspace analysis is the analysis of the

components present in that gas.
heat

GC
Thermal Desorption
 Purge and Trap

• Way to measure dilute samples by concentration of

constituents
• Trap constituents under low temperature
• Heat trap to release constituents and send to GC column

Trap

Adsorbents: graphitized carbon,

alumina, silica gel, activated
carbon, etc.
 Column
flow
sample
meter
oven

data
carrier system
injector column detector
gas /readout
/display

reference gas
 Columns

Packed column.
Typically used
in HPLC,
seldom in GC,
they offer high
capacity but
poor resolution.
Typically used in GC:
rapid equilibration is accomplished by decreasing
stationary phase thickness and reducing column
diameter (Cu term is reduced  good resolution)
Open Tubular versus packed Columns
In OTC we generally find:
1. higher resolution
 greater length (up to 100 times) possible, and so more theoretical plates
possible for OTC.
2. shorter analysis time
 higher flow rates possible for OTC (less resistance).
3. lower sample capacity  not useful for preparative purposes when
compared to packed columns.
Effect of decreased thickness of
the stationary phase in GC
Decreased retention times

Decreasing Decreased sample

thickness of capacity (less
stationary phase
thickness, less material)

Increased resolution*
(less thickness, less material, so
less diffusion).
Choosing the Column
 The (Kovats) retention index

Methan Cn+4H2n+4 unknown Cn+4H2n+4

(void)
• Programmable Oven
• Isothermal- run at one constant temperature
• Temperature programming (Temperature Gradient ) -
Start at low temperature and gradually ramp to higher
temperature. Temperature of a column is raised during
the separation to increase solute vapor pressure. as
temperature increases, retention time decreases and
peaks are sharpening

Isothermal: 150°C Programmed T: 50°C- 250°C

 Detectors
flow
sample
meter
oven

data
carrier system
injector column detector
gas /readout
/display

reference gas
1. Thermal Conductivity Detector (TCD)
2. Flame Ionization Detector (FID)
3. Nitrogen-phosphorus Detector
4. Electron Capture Detector (ECD)
5. Mass Spectrometers
 Thermal Conductivity Detector (TCD)
Operates on the changes in the thermal conductivity of the
gas stream brought about by the presence of analyte
molecules. He is the carrier gas most often used with this
detector because it has a high thermal conductivity.
+ truly universal detector
applicable to the detection of any compound in GC
+ non-destructive
useful for detecting compounds from preparative-scale columns
useful in combination with other types of GC detectors

- detect mobile phase impurities

- sensitive to changes in flow-rates
- limit of detection
~ 10-7 M
much higher then other GC detectors
Flame Ionization Detector (FID)
• eluate is burned in a mixture of H2 and
air. Only 1 out of 100,000 carbon atoms
produce ions, but the number of ions is
strictly proportional to the number of
carbon entering the flame !.
• From carbon atoms (except C=O and
COO) CH radicals are generated, that
can be ionized in the detector flame:
*CH + *O  CHO+ + e –

• if voltage is applied between the

electrodes, current (=movement of
electrons) can be recorded.
• response is proportional to number of
molecules
• + linear range is 7 orders of magnitude
+ generally sensitive to hydrocarbons
– insensitive to non-hydrocarbons
 Nitrogen-Phosphorus Detector (NPD)
- used for detecting nitrogen- or phosphorus containing compounds
- also known as alkali flame ionization detector or thermionic detector

- same basic principal as FID

- measures production of ions when a solute is burned in a flame
- enhances the formation of ions from nitrogen- and phosphorus-
containing compounds
Alkali metals
FPD（Flame Photometric Detector）

S: 393 nm
P: 526 nm Rb* + CN →Rb++ CN-
 Electron Capture Detector (ECD)
- radiation-based detector
- selective for compounds containing electronegative
atoms, such as halogens

N2 + β ray → N2+ + e-
PCB + e- → PCB- (slow)
- electrons go to collector electrode where
they produce a current
- compounds with electronegative atoms
capture electrons, reducing current
 Derivatization
What
Derivatization is the process of chemically modifying a
compound to produce a new compound which has
properties that are suitable for analysis using a GC.

Why
● To permit analysis of compounds not directly
amenable to analysis due to, for example, inadequate
volatility or stability
● Improve chromatographic behavior or detectability
 Derivatization
Trifluoroacetylation: TFA (Trifluoroacetic Acid)
amine: R-NH2 + (CF3CO)2O
→R-NHCOCF3 + CF3COOH
alcohol: R-OH + (CF3CO)2O
→R-OCOCF3 + CF3COOH

Trimethylsilylation: TMS (Trimethylchlorosilane)

alcohol: R-OH + (CH3)3SiCl
→R-OSi(CH3)3 +HCl
carboxylate: R-COOH + (CH3)3SiCl
→RCOOSi(CH3)3 +HCl