1

Gas Chromatography
A.) Introduction:

Gas Chromatography (GC) - chromatographic technique where the mobile phase is a gas.

GC is currently one of the most popular methods for separating and analyzing compounds. This is due
to its high resolution, low limits of detection, speed, accuracy and reproducibility.

GC can be applied to the separation of any compound that is either naturally volatile (i.e., readily goes
into the gas phase) or can be converted to a volatile derivative. This makes GC useful in the
separation of a large number of small organic and inorganic compounds.

B.) Equipment:

A simple GC system consists of:

1. Gas source (with pressure and flow regulators)
2. Injector or sample application system
3. Chromatographic column (with oven for temperature control)
4. Detector & computer or recorder
 A typical GC system used is shown below (a gas chromatograph)

Carrier gas: He (common), N2, H2

Pinlet 10-50 psig
Flow = 25-150 mL/min packed column
Flow = 1-25 mL/min open tubular column
Column: 2-50 m coiled stainless steel/glass/Teflon
Oven: 0-400 °C ~ average boiling point of sample
Accurate to <1 °C
Detectors: FID, TCD, ECD, (MS)
3
4 GC Theory

 An inert gas such as helium is passed through the column as a carrier gas and is the
moving phase. A sample is injected into a port which is much hotter than the column and
is vaporized. The gaseous sample mixes with the helium gas and begins to travel with
the carrier gas through the column. As the different compounds in the sample have
varying solubility in the column liquid and as these compounds cool a bit, they are
deposited on the column support. However, the column is still hot enough to vaporize the
compounds and they will do so but at different rates since they have different boiling
points. The process is repeated many, many times along the column. Eventually the
components of the injected sample are separated and come off of the column at different
times (called "retention times").
5

 There is a detector at the end of the column which signals the change in the nature of
the gas flowing out of the column. Recall that helium is the carrier gas and will have a
specific thermal conductivity, for example. Other compounds have their own thermal
conductivities. The elution of a compound other than helium will cause a change in
conductivity and that change is converted to an electrical signal. The detector, in turn,
sends a signal to a strip chart recorder or to a computer. Detectors come in several
varieties, for example, thermal detectors, flame-ionization and electron capture
detectors.
6 Theory

 In order to understand GC, need to focus on the general

principles of separations which has its roots in solvent
extraction. The theory of solvent extraction are used to
explain all forms of chromatography i.e. HPLC, LC, GC,
EP, and TLC.
 Consider two solvents S1 and S2 and solute X that is in
S1. The partition between the two phases.
 X(S1)  X(S2);
7 GC Theory

 Partition coefficient K is an equilibrium constant is K=

[X]S1/[X]S2.
 Suppose that solute X in V1 (water) is extracted with V2 (CCl4).
 Let m be the moles of X in the system and let q be the fraction of X
remaining in phase 1 at equilibrium.
 The molarity in phase 1 (water) is therefore qm/V1.
8 GC Theory

 The fraction of total solute transferred to phase 2 (CCl4) is (1-q)

and the molarity in phase 2 is
 (1-q) m / V2. Then
 K = ((1-q) m / V2)/ q m V1.
 For GC:
 K = Cs/Cm where Cs is the concentration in the stationary phase
(column) and Cm is the concentration in the mobile phase (gas).
9 Suppose I2 in H2O is 1 and I2 in CCl4 is 0.

 After shaking and letting settle, I2 in H2O is 0.0014 and I2 in CCl4 is

0.9986. Therefore
 K = 7 x 102. Fraction (q) remaining after the 1st extraction is:
 q = V1/(V1+KV2). Fraction remaining after n extractions is
 qn = (V1/(V1+KV2))n. It is more efficient to do several small
extractions than one large one. This extraction can be used to
describe GC where the liquid is the mobile phase and the column is
the stationary phase. Each extraction is a theoretical plate.
10 Theory of separation Water + I2

Water + I2 Water + I2

CCl4 CCl4 + I2

CCl4
+ I2

Start Shake 1 Shake 2

11 Theory GC

 As the gas moves the solute (analyte) through and over the stationary phase, the
solute will be in equilibrium with the gas and the solid phase. Since there is a
mobile phase, the separation will appear as a chromatogram showing the
separation of the analytes.
 GC Theory
12
As the gas moves the solute (analyte) through and over the stationary phase, the
solute will be in equilibrium with the gas and the solid phase. Since there is a
mobile phase, the separation will appear as a chromatogram showing the
separation of the analytes.

Analyte

To detector

Column + packing

Time
 Separations
13

To detector

Time 1 Time 2 Time 3

14 Capacity Factor (k’)
• While inside the column, a retained component spends part of its time on the stationary
phase and part time in the mobile phase
 • When in the mobile phase, solutes move at the same speed as the mobile phase
 • this means that all solutes spend the same amount of time in the mobile phase
(to)
 • the amount of time the solute spends on the stationary phase is equal to tR-
to(adjusted retention time, t’R)
 •the ratio t’R/ to is the capacity of the column to retain the solute (k’)

tR
to t’ R

k’ = (tr - t0) / t0
k’ = (t’r ) / t0
.
Inject

Unretained
Solute
15 GC Process
 Column Efficiency (N)
16
 Solutes are placed on an GC column in a narrow band
 • Each solute band spreads as it moves through the column due
to diffusion and mass transfer affects
 • The later eluting bands will spread more

 • Peak shape follow a Gaussian distribution

to
t1
t2

Band spreading eventually causes peaks to merge into the baseline. We want to
minimize band spreading as much as possible.
17 Chromatogram nomenclature
18 GC Peak parameters
19 Typical separation
20 GC General
 The chromatogram shows the order of elution (order of
components coming off the column), the time of elution
(retention time), and the relative amounts of the components
in the mixture.

 The order of elution is related to the boiling points and

polarities of the substances in the mixture. In general, they
elute in order of increasing boiling point but occasionally the
relative polarity of a compound will cause it to elute "out of
order". This is analyzing your sample.
21 Elution Order

  Compound  Boiling Point (0C)

  pentane  36
  hexane   69
  cyclohexane  80
  iso-octane 99
  toluene  110
  4-methyl-2-pentanone  117
  octane  126
22 GC Chromatogram
• The observed elution pattern appears below. Notice the
reversed elution of toluene and 4-methyl-2-pentanone.
23 Column temperature
 For precise work, column temperature must be controlled to within tenths of a
degree. The optimum column temperature is dependant upon the boiling point of
the sample. As a rule of thumb, a temperature slightly above the average boiling
point of the sample results in an elution time of 2 - 30 minutes. Minimal
temperatures give good resolution, but increase elution times. If a sample has a
wide boiling range, then temperature programming can be useful. The column
temperature is increased (either continuously or in steps) as separation proceeds.
24 Mobile Phase (gas)
25 GC Under the hood
26 GC Column and Oven
27 Typical GC (dual column)
28 Sample Injections
 Next, the sample injection system. Here it is important that
the sample be injected onto the column as a plug and of a
suitable size. Also, the injector should provide consistent and
reproducible injections.
 The micro-syringe is used to load the sample onto the
column. The syringe should be clean and accurate and gas
tight. The syringe is injected through a rubber septum. The
septum should be replaced after many injections to insure gas
tightness onto the column. An auto sampler can be used to
inject the samples. Typical volumes range from 0.2 to 20 Ls.
With capillary columns it is necessary to use a splitter. (See
Figure.) A suitable solvent is also necessary for the proper
separations and injections.
29 GC Injector
30 Carrier Gas

 This is the mobile phase and should be pure gas so as not to react with the column or
analyte. Gas is usually He, Ar, N2, or H2. Choice will depend on the type of detector used.

He and H2 give better resolution (smaller plate height) than N2. Pressure is also important
and as expected the system comes with regulators. Can you find where in GC equations
that are dependent on pressure?
31 Columns

 The column is the most important component of GC. Here is where

the separations take place. All the various equations we discussed
above are dependent on properties of the column. There are four
types of columns: wall-coated open tubular (WCOT), support
coated open tubular (SCOT), micropacked, fused silica open tubular
(FSOT), and packed column. The FSOT column is the most
flexible. Open tubular is also capillary. Particle size is important
because the efficiency of GC column increases rapidly with
decreasing particle size of the packing material.
32 Column

 The column sits in a temperature controlled environment

that is 0.50. Temperature is very important in GC.
 Normally, one does a temperature program to get the
various analytes off the column for better separations
(resolutions).
33 Columns

 There are two general types of column, packed and capillary (also known as open tubular).
Packed columns contain a finely divided, inert, solid support material (commonly based on
diatomaceous earth) coated with liquid stationary phase. Most packed columns are 1.5 -
10m in length and have an internal diameter of 2 - 4mm.
 Capillary columns have an internal diameter of a few tenths of a millimeter. They can be
one of two types; wall-coated open tubular (WCOT) or support-coated open tubular
(SCOT). Wall-coated columns consist of a capillary tube whose walls are coated with
liquid stationary phase. In support-coated columns, the inner wall of the capillary is lined
with a thin layer of support material such as diatomaceous earth, onto which the stationary
phase has been adsorbed. SCOT columns are generally less efficient than WCOT columns.
Both types of capillary column are more efficient than packed columns.
 In 1979, a new type of WCOT column was devised - the Fused Silica Open Tubular
(FSOT) column;
34 FSOT column
35 Capillary column
 C.) Mobile Phase:
36 GC separates solutes based on their different interactions with the mobile and stationary
phases.
- solute’s retention is determined mostly by its vapor pressure and volatility
- solute’s retention is controlled by its interaction with the stationary phase
- gas mobile phase has much lower density
‚decreased chance for interacting with solute
‚increased chance that solid or liquid stationary phase interacts with solute
 C.) Mobile Phase:
37 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

 C.) Mobile Phase:
Carrier Gas or Mobile phase does not affect solute retention, but does affect:
38
1.) Desired efficiency for the GC System
- low molecular weight gases (He, H2)  larger diffusion coefficients
- low molecular weight gases  faster, more efficient separations

2.) Stability of column and solutes

- H2 or O2 can react with functional groups on solutes and stationary
phase or with surfaces of the injector, connections and detector

3.) Response of the detector

- thermal conductor requires H2 or He
- other detectors require specific carrier gas
 D.) Stationary Phases:

Stationary phase in GC is the main factor determining the selectivity and retention
39 of solutes.

There are three types of stationary phases used in GC:

Solid adsorbents
Liquids coated on solid supports
Bonded-phase supports

1.) Gas-solid chromatography (GSC)

- same material is used as both the stationary phase and support material
- common adsorbents include:
‚ alumina
‚ molecular sieves (crystalline aluminosilicates [zeolites] and clay)
‚ silica
‚ active carbon

Magnified Pores in activated carbon

40 Gas-solid chromatography (GSC):

advantages:
- long column lifetimes
- ability to retain and separate some compounds not easily resolved by other
GC methods
‚ geometrical isomers
‚ permanent gases

disadvantage:
- very strong retention of low volatility or polar solutes
- catalytic changes that can occur on GSC supports
- GSC supports have a range of chemical and physical environments
‚ different strength retention sites
‚ non-symmetrical peaks
‚ variable retention times
 2.) Gas-liquid chromatography (GLC)
- stationary phase is some liquid coated on a solid support
41 - over 400 liquid stationary phases available for GLC
‚ many stationary phases are very similar in terms of their retention properties
- material range from polymers (polysiloxanes, polyesters, polyethylene glycols) to
fluorocarbons, molten salts and liquid crystals

Based on polarity, of the 400 phases available only 6-12 are needed for most separations.
The routinely recommended phases are listed below:

Chemical nature of Max. McReynolds’ constants

Name polysiloxane temp. x’ y’ z’ m’ s’
SE-30 Dimethyl 350 14 53 44 64 41

Dexsil300 Carborane-dimethyl 450 43 64 111 151 101

OV-17 50% Phenyl methyl 375 119 158 162 243 202

OV-210 50% Trifluoropropyl 270 146 238 358 468 310 Higher the number the
OV-225 25% Cyanopropyl- 250 238 369 338 492 386 higher the absorption.
25% phenyl
Silar-SCP 50% Cyanopropyl- 275 319 495 446 637 531
50% phenyl
SP-2340 75% Cyanopropyl 275 520 757 659 942 804

OV-275 Dicyanoallyl 250 629 872 763 1106 849

McReynolds’ constants based on retention of 5 standard “probe” analytes

– Benzene, n-butanol, 2-pentanone, nitropropanone, pyridine
 Preparing a stationary phase for GLC:
- mixture of the desired liquid phase and solvent is made with a solid support
‚ solid support is usually diatomaceous earth (fossilized shells of
42
ancient aquatic algae (diatoms), silica-based material)
- solvent is evaporated off, coating the liquid stationary phase on the support
- the resulting material is then packed into the column

disadvantage:
- liquid may slowly bleed off with time
‚ especially if high temperatures are used
‚ contribute to background
‚ change characteristics of the column with time
 3.) Bonded-Phase Gas chromatography
- covalently attach stationary phase to the solid support material
43 - avoids column bleeding in GLC
- bonded phases are prepared by reacting the desired phase with the surface of a silica-
based support
reactions form an Si-O-Si bond between the stationary phase and support
or
reactions form an Si-C-C-Si bond between the stationary phase and support
- many bonded phases exist, but most separations can be formed with the following
commonly recommended bonded-phases:
‚ Dimethylpolysiloxane
‚ Methyl(phenyl)polysiloxane
‚ Polyethylene glycol (Carbowax 20M)
‚ Trifluoropropylpolysiloxane
‚ Cyanopropylpolysiloxane

CH3 CH3 C6H5

H H

O Si O Si O Si
HO C C O H

CH3 CH3 C6H5

n n m H H n

advantages:
- more stable than coated liquid phases
- can be placed on support with thinner and more uniform thickness than
liquid phases
 E.) Support Material:

44 There are two main types of supports used in GC:

Packed columns
‚ large sample capacity
‚ preparative work
Capillary (open-tubular) columns
‚ higher efficiency
‚ smaller sample size
‚ analytical applications
 F.) Elution Methods:

45 A common problem to all chromatographic techniques is that in any one sample there may
be many solutes present, each retained by the column to a different degree:

Best separation and limits of

detection are usually
obtained with solutes with k’
values of 2-10

Difficult to find one condition

that elutes all solutes in this
k’ range  general elution
problem

Gradient elution - change column condition with time which changes retention of solutes to
overcome general elution problem
 Temperature Programming – changing the temperature on the column with time to simulate
gradient elution in GC since a solute’s retention in GC is related to its volatility.
46
Comparison of a GC separation using isothermal conditions and temperature programming:

ISOTHERMAL
Column temp. 120oC

Programmed temp.
(30oC to 180oC) (5o/min)

Temperature programming is usually done either in a stepwise change, a linear change or a

combination of several linear changes. A single linear change or ramp is the most common
 G.) GC Detectors:

47 The following devices are common types of GC detectors:

1. Thermal Conductivity Detector (TCD)
2. Flame Ionization Detector (FID)
3. Nitrogen-phosphorus Detector (NPD)
4. Electron Capture Detector (ECD)
5. Mass Spectrometers (discussed later in the course)

The choice of detector will depend on the analyte and how the GC method is being
used (i.e., analytical or preparative scale)

Detector Application Approx. Sensitivity Notes

Cost
TCD everything $3-5 10’s of nanograms Not very sensitive, easy to operate, only one
gas required
FID hydrocarbons $4-6 Sub-nanogram Very linear, relatively easy to operate,
required fuel gasses, not sensitive to all
NPD Nitrogen/sulfur $10-12 Low-picograms Very selective, hard to operate, required fuel
gases
ECD Halogenated, nitro $6-9 Low-picograms Very sensitive, radiation source, not very
linear, selective, two gases
MS Almost everything $40 Depends on operation Sensitive, requires pump system, fairly
complicated requires cleaning
 1.) Thermal Conductivity Detector (TCD)
48
- katherometer or hot-wire detector
- first universal detector developed for GC

Process
- measures a bulk property of the mobile phase leaving the column.
- measures ability to conduct heat away from a hot-wire (i.e., thermal conductivity)
- thermal conductivity changes with presence of other components in the mobile phase
 Design
- based on electronic circuit known as a Wheatstone bridge.
49 - circuit consists of an arrangement of four resistors with a fixed current applied to them.
- thermal conductivity changes with presence of other components in the mobile phase.
- the voltage between points (+) and (-) will be zero as long as the resistances in the
different arms of the circuit are properly balanced

-one resistor in contact with mobile

phase leaving column

-another in contact with reference

stream of pure mobile phase

as solute emerge from column:

change in thermal conductivity  change in amount of heat removed from resistor 
change in resistor’s temperature and resistance  change in voltage difference between
points (+) and (-).
 Considerations
- mobile phase must have very different thermal conductivity then solutes being
50 separated.
- most compounds separated in GC have thermal conductivity of about 1-4X10 -5.
- H2 and He are carrier gases with significantly different thermal conductivity values.
- H2 reacts with metal oxides present on the resistors, so not used

advantages:
- 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

disadvantage:
- detect mobile phase impurities
- sensitive to changes in flow-rates
- limit of detection
‚ ~ 10-7 M
‚ much higher then other GC detectors
 2.) Flame Ionization Detector (FID)
- most common type of GC detector
51
- “universal” detector capable of measuring the presence of almost any organic and many
inorganic compound

Process
- measures the production of ions when
a solute is burned in a flame.

- ions are collected at an electrode to

create a current
 2.) Flame Ionization Detector (FID)

52
advantages:

- universal detector for organics

‚ doesn’t respond to common inorganic compounds

- mobile phase impurities not detected

- carrier gases not detected

- limit of detection: FID is 1000x better than TCD

- linear and dynamic range better than TCD

disadvantage:

- destructive detector
 3.) Nitrogen-Phosphorus Detector (NPD)
53
- used for detecting nitrogen- or phosphorus containing compounds
- also known as alkali flame ionization detector or thermionic detector

Process
- same basic principal as FID
Alkali Bead

- measures production of ions when a solute

is burned in a flame

- ions are collected at an electrode to

create a current

- contains a small amount of alkali metal

vapor in the flame

- enhances the formation of ions from

nitrogen- and phosphorus- containing compounds
 3.) Nitrogen-Phosphorus Detector (NPD)
54
advantages:
- useful for environmental testing
‚ detection of organophosphate pesticides

- useful for drug analysis

‚ determination of amine-containing or basic drugs

- Like FID, does not detect common mobile phase impurities or carrier gases

- limit of detection: NPD is 500x better than FID in detecting nitrogen- and
phosphorus- containing compounds

- NPD more sensitive to other heterocompounds, such as sulfur-, halogen-, and

arsenic- containing molecules

disadvantage:
- destructive detector

- NPD is less sensitive to organic compounds compared to FID

 4.) Electron Capture Detector (ECD)

55 - radiation-based detector
- selective for compounds containing electronegative atoms, such as halogens

Process
- based on the capture of electrons by
electronegative atoms in a molecule

- electrons are produced by ionization of the

carrier gas with a radioactive source
‚ 3H or 63Ni

- in absence of solute, steady stream of

these electrons is produced

- electrons go to collector electrode where

they produce a current

- compounds with electronegative atoms

capture electrons, reducing current
 4.) Electron Capture Detector (ECD)
56
advantages:
- useful for environmental testing
‚ detection of chlorinated pesticides or herbicides
‚ detection of polynuclear aromatic carcinogens
‚ detection of organometallic compounds

- selective for halogen- (I, Br, Cl, F), nitro-, and sulfur-containing compounds

- detects polynuclear aromatic compounds, anhydrides and conjugated

carbonyl compounds
57 Applications
 Pesticides
58
Analysis of pesticide residues in soil, water, and food is crucial for maintaining safe
levels in the environment. The PDD in the ECD mode is highly selective for
monitoring electron capturing compounds such as chlorinated pesticides and other
halogens. This chromatogram illustrates the sensitivity of the non-radioactive
PDECD for such compounds.

Retention time (sec)

59 Food analysis

 Analysis of foods is concerned with the assay of lipids, proteins, carbohydrates,

preservatives, flavours, colorants and texture modifiers, and also vitamins,
steroids, drugs and pesticide residues and trace elements. Most of the components
are non-volatile and although HPLC is now used routinely for much food
analysis, GC is still frequently used. For examples, derivatization of lipids and
fatty acid to their methyl esters(FAMEs), of proteins by acid hydrolysis followed
by esterification (N-propyl esters) and of carbohydrates by silylation to produce
volatile samples suitable for GC analysis.
60 GC Food

 GC quality control analysis of food products can confirm the presence and quantities of the
analytes  For example, fruits, fruit derived foodstuffs, vegetables and soft drinks, tea and
coffee, were analyzed for their polybasic and hydroxy acid contents (citric, maleic acids) as
TMS derivatives.

 All food and beverage products on sale today must be carefully assayed for contamination
with pesticides, herbicides and many other materials that are considered a health risk. The
analysis of food involves separating and identifying very complex mixtures, the components
of which are present at very low concentrations. GC is the ideal technique for use in food
and beverage assays and tests. Furthermore, the origin of many herbs and spices can often
be identified from the peak pattern of the chromatograms from their head space analysis.
61 Food and Cancer

 Chemicals that can cause cancer have a wide variety of

molecular structures and include hydrocarbons, amines,
certain drugs, some metals and even some substances
occurring naturally in plants and molds. In this way, many
nitrosamines have carcinogenic properties and these are
produced in a number of ways such as cigarette smoke.
GC can be used to identify these nitro-compounds in trace
quantities.
 Drugs
62

 There are still numerous GC applications involving both quantitative and qualitative
identification of the active components and possible contaminants, adulterants or
characteristic features which may indicate the source of the particular sample. Forensic
analysis frequently users GC to characterize drugs of abuse, in some cases the characteristic
chromatographic fingerprint gives an indication of the source of manufacture of the sample
or worldwide source of a vegetable material such as cannabis.

 Analytical procedures, chromatographic methods and retention data are published for over
600 drugs, poisons and metabolites. These data are extremely useful for forensic work and
in hospital pathology laboratories to assist the identification of drugs.
63 Pyrolysis gas chromatography

 Pyrolysis GC (PGC) is used principally for the identification

of non-volatile materials, such as plastics, natural and
synthetic polymers, drugs and some microbiological materials.
The thermal dissociation and fragmentation of the sample
produces a chromatogram which is a fingerprint for that
sample. The small molecules produced in the pyrolysis
reaction are frequently identified using a GC-MS system and
information on molecular structure for identification is also
obtained.
64 Metal chelates and inorganic materials

 Although inorganic compounds are generally non-volatile, GC

analysis can be achieved by converting the metal species into
volatile derivatives. Only some metal hydrides and chlorides
have sufficient volatility for GC. Organometallics other than
chelates, which can be analyzed directly, include boranes,
silanes, germanes, organotin and lead compounds.
65 Environmental analysis
 Environmental pollution is an age-old trademark of man and in recent years as technology has
progressed, populations have increased and standards of living have improved. So the demands
on the environment have increased, with all the attendant problems for the ecosystems.

 Combustion of fossil fuel, disposal of waste materials and products, treatment of crops with
pesticides and herbicides have all contributed to the problems. Technological developments
have enabled man to study these problems and realize that even trace quantities of pollutants
can gave detrimental effects on health and on the stability of the environment.

 There is a vast amount of literature on the use of GC for studying a wide variety of these
problems.