Introduction to

Gas
Chromatography

Chapter 29
A Quick Historical
Perspective
• 1903 Adsorption chromatography
• 1952 Gas chromatography (GLC)
• 1968 High performance liquid
chromatography (HPLC)
 HPLC GC
• Mobile phase changes • Mobile phase is constant
• Constant temperature • Increasing temperature
• Compounds partition • Compounds partition from
from the mobile phase the mobile phase based
based on solubility. on volatility.
• Elution is generally • Elution is generally
time or volume temperature dependent
dependent
 Sample Injected

Low Temperature

High Temperature
Typical Volatile Profile of Strained Carrots
(Common Terpenoids Marked in Blue)
Sample Preparation

How do we obtain a volatile sample from

foods?
Sample Preparation
1) Headspace Methods
2) Distillation Methods
3) Solvent Extraction
4) Solid-Phase Microextraction
 Direct Headspace
In a closed container, take a sample of the air above the
sample and inject it.

While direct injection of gasses can be used, many volatile

analytes are often in too low of a concentration to
be quantified using most common detectors.
 Dynamic Headspace
Typically the sample is placed in a closed container and a flow
of inert gas such as nitrogen is used to purge the headspace
onto an adsorbent or a cryogenic trap.

The trap can be eluted with solvent, the

solvent can be concentrated and injected.
 Solvent Extraction
Blend sample and mix with
non-miscible solvent. (a solvent
that doesn’t mix with water).

Centrifuge to unmix solvent

and food sample
Solvent Extraction
Separate the solvent containing
the analytes of interest and
concentrate by evaporation
for injection into GC.
 Distillation Methods

Produces a dilute aqueous Food

sample which is not the Sample
best for GC. Sample can
be re-extracted with solvent.

Sample
 Solid Phase Microextraction

Sampling devices (manual and autosampler)

consist of a coated silica fiber inside a
hollow needle. The coating on the fiber
adsorbs and concentrates volatiles from
the headspace inside the sample container.
The needle protects the fiber which can be
inserted through the septa of the GC injector
for direct analysis.

Probably the easiest method of sample prep.

Hardware and Columns
• Mobile Phase: He, Ar, N2, H2
• Injection Port: Rubber septum barrier
(usually maintained at a higher temperature
than the boiling point of the least volatile
component in the sample mixture)…why?
• Column: (fused silica with a thin coating of
stationary phase on the inner surface)
• Oven: Thermostat controlled forced air
oven
• Detector: Many types
• Data System
 Split/Splitless Injector
Splitless Injection,
(where the split vent is closed)
attempts to transfer all of the
sample to the column and is
used for trace analysis.

Split Mode,
only a small portion
(maybe 1-10% of the sample
moves into the column, and
the rest is sent to waste. This
is used when the analytes are
in high concentration and would
overload the column.
Sample is injected through the
septum with a syringe.
The oven

Inside here
Column
Question:

• The injection port is at a HIGH temperature

• 250oC is common.

What happens when the hot, volatized sample hits a

“cold” (50oC) GC column?
• The volatile compounds condense at the front end
of the column. Raising the temperature of the
column allows for the separation of the
compounds as they boil away at different
temperatures.

• If two compounds have the same volatility, the

compound with the least affinity for the stationary
phase will volatilize first.
Gas chromatography separates
molecules mostly based on:

1. Volatility (Oven)
2. Polarity (Column)
 Oven Temperature Program
Increase Temp. Linearly
or Stepwise
Temperature

Time
 COLUMNS
• GC column nomenclature can be confusing.
• Carbowax, DB-1, DB-5, PDMS …..
• Columns are often referred to by their polarity, like most
things with chromatography. The most non-polar
stationary phase is polydimethyl siloxane (PDMS).
• Polarity of a column is increased by adding phenyl
groups to PDMS (1% = DB-1; 5% = DB-5).
• For more polar analytes, polyethylene glycol (carbowax)
is used as the stationary phase
 COLUMNS
Compound Polarity Max Temp oC Column ID Manufacturer
Poly(methyl siloxane) Low 300-350 HP-1 Agilent
AT-1 Alltech
DB-1, SE-30 J&W
OV-1 Ohio Valley
ZB-1 Phenomenex
RTx-1 Restek
BP-1 SGE
SPB-1 Supelco
CP-Sil 5 CB Varian
95% Dimethyl, 5%phenyl Low 300 HP-5 Agilent
Poly(methyl siloxane) AT-5, EC-5 Alltech
DB-5, SE-54 J&W
OV-5 Ohio Valley
ZB-5 Phenomenex
RTx-5 Restek
BP-5 SGE
SPB-5,MDN-5 Supelco
CP-Sil 8 CB Varian
Polyethylene glycol Medium 250 HP-20M Agilent
AT-Wax Alltech
DB-Wax J&W
Carbowax 20M Ohio Valley
ZB-Wax Phenomenex
Stabilwax Restek
BP20 SGE
Supelcowax 10 Supelco
CP-Wax 52 CB Varian

About a dozen different types in common usage.

 GC Applications with Food
• Analysis of foods is concerned with the assay of lipids,
proteins, carbohydrates, preservatives, flavors, colorants,
vitamins, steroids, drugs, and pesticide residues.
• Most of the components are non-volatile (thus the use of
HPLC) but with modification, GC can be effectively used.
• Derivatization of lipids and fatty acid to their methyl esters
• Proteins are acid hydrolyed followed by esterification (N-
propyl esters)
• Carbohydrates derivatized by silylation to produce a volatile
compound