Gas Chromatography – Harris

Chapter 24
Gaseous Mobile Phase, Solid or
Liquid Stationary Phases

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Schematic & Major Components (fig

24-1)

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 Carrier
Gas
Reservoir
Gas Flow
Make-up Gas
(optional)

Injection Port

Sample
Introduction

Separation
column

Injection Port Oven Detector Oven

Temperature T1 Temperature T3
Column Oven
Temperature T2

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Summary of GC components.
z Injection port – Syringe containing sample is introduced through
a septum, injection port oven temperature heated to temperatures
that ensures fast volatilization of sample, i.e. above the b.p. of all
sample components, usually 275 0C. Keep in mind these terms:
split and splitess injection

z Carrier Gas – Sweeps analyte to separation column. Usually, He,

or N2, sometimes H2.

z Column – Either packed (old) or capillary (newer) columns. Each

type requires its own set of plumbing components (threads, etc.)
Interestingly all GC and HPLC plumbing components in the
English system of measurements.

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Summary of GC components.
z Column T – May be below b.p. of sample components, but high
enough to keep a significant quantity in the gas phase.
Temperature may be ramped up to get separation based on b.p.
differences of components along with chromatographic
separation.

z Makeup Gas – Sometimes required since detector volume is too

large for carrier gas flow. Remember that makeup gas is usually
required for capillary columns.

z Detector – Ideally, want universal detector, sensitive to anything

that passes by. Wide dynamic range, low D.L. the usual wants.
We want to keep all sample components volatilized, detector
oven temperature usually 300 0C.

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Effects of the Column and Mobile

Phase Flow in GC.
z Why are modern GC based on a capillary column?

z Back to the van Deemter Eqn. H = A + B/u + Cu

z Remember, u = flow rate,

z A = multiple paths
z B/u = longitudinal diffusion effects
z Cu = MT effects

z We want to minimize H as much as possible.

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“Older” packed columns

z usually 1/8” (3.2 mm OD, 2.2 mm I.D.)

diameter, 1 – 2 m length
z design impedes gas flow
z max flow rate about 1 mL/min or 8
cm/min.

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“Newer’ capillary columns
z 0.25 mm inner diameter.
z major point, less restricted flow path.
z flow rate about 20 ml/min or 20 cm/min

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We want to minimize H as much as

possible.
z H = A + B/u + Cu

z Which of the above affects H the most in GC?

• B/u effects!
z Diffusion coefficients are large in the gas phase.
σ = 2 D uL
z Simply increasing the flow rate partially
addresses the B/u effects in GC

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Other major factors that influence the performance of
capillary columns relative to packed ones are evident
in the table below.
Typical 1/8 “ Typical Capillary Comments
packed
I.D. 2.2 mm 0.25 mm No packing material less
0.1-0.53 mm restricted flow
df 5 µm 0.25 µm MT for s.p. part of Cu
f (k ' )d 2f
Cs α
Ds

L 1-2 m 10-60 m HN = L
N 4,000 180,000

Advantages •Lower cost •Higher sep.

•Easy to make efficiency
•Larger •Faster sep.
samples •Better for complex
mixtures

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Other advantages of tubular over packed GC

columns:

z No “A” term, straight path.

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Example of the separation efficiency of capillary vs.
packed column.

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GC stationary phases.
z Older packed columns – uniform silica particles (150-250 µm)
required to ensure uniform path lengths (the “A” term in the van
Deemter eqn.) Surfaces are chemically modified (see below). The
columns themselves were either glass or stainless steel.

z Capillary columns – fused silica which like the particles in the

packed column require chemical modification (below).The
stationary phase surface (silica) is a hydroylated surface. This
caused problems with nonpolar stationary phases.

z When polar or mildly polar species partition into the s.p. they
stand a good chance of being trapped, causing a excessive band-
broadening beyond what is expected from van Deemter
considerations.

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Trapped polar solute

Mobile Phase

Stationary Phase

OH OH OH OH OH OH OH OH

Silica Support

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Surface modification with trimethylchlorosilane
reduces surface polarity.

CH3
H3C CH3
Si
Cl
CH3
+ H3C CH3
Si
OH OH OH O + HCl
O Si O Si O Si O Si O
O O O O

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Effects can be dramatic:

z A = EtOH
z B = methylethyl
ketone
z C = benzene
z D = cyclohexane

z Chromatogram 1
= unmodified

z Chromatogram 2
= silinazied

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Liquid stationary phases

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Important Notes:

• All columns have a Tmax, never exceed this!

Thermal degradation, or s.p. simply boils
off.

• Most s.p. are air sensitive and easily oxidize

at high temperatures.

• Capillary columns cost $150-600 (2002

dollars)

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Effect of Stationary Phase Polarity on
Retention Times.

z #1 n-heptane
z #2 tetrahydrofuran
z #3 2-butanone
z #4 n-propanol

z We can reason this order by

considering the polarity of the
analytes and of the s.p.

z In GC the s.p. characteristics

have the largest effect on the
order of elution.

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Column Temperature and

Temperature Programming.
z The oven T must be controlled to within ±0.5
0C from RT to 400 0C. A fan circulates air
throughout the oven ensuring uniform heating.

z Constant temperature isothermal separations

are good for simple mixtures.

z Typically T programming is required. Slowly

ramping T throughout the separation provides
a basis for the separation of sample
components based on BP.

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Isothermal vs. Temp Programming
z The more
volatile cpds
elute very
close together
under
isothermal
conditions
(upper).

z Ramping T
from 50 to 250
oC at 8 oC/min
separates out
those volatile
cpds (lower).

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How does T programming work?

• T programming is an example of the control of the capacity

t − tm
factor: k A = t
' r
which in turn controls the resolution term:
m

⎛ α − 1 ⎞⎛ k B ⎞
'
N
Rs = ⎜ ⎟⎜⎜ ⎟
' ⎟
4 ⎝ α ⎠⎝ 1 + k B ⎠

• Note that T programming does not change the order of

elution.

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Carrier Gases
• O2 is usually avoided since it will oxidize the s.p.

• 3 most common gases N2, H2, He.

• Each have very different Hmin characteristics.

• The lighter gases He and H2 require faster analysis flow rates

20-50 cm/min.

• Diffusion in He and H2 causes more band-broadening in m.p.

• Greater MT effects decrease band-broadening at faster flow
rates
• H = A + B/u + Cu

• N2 Hmin occurs at about 10 cm/min.

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Sample Injection

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Injection Port for Modern GC’s

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GC Injection Syringe
z It is important to rapidly vaporize the sample.

z Slow vaporization increases band broadening, by

increasing the sample “plug”.
z
Injection port temperature is usually held 50 0C higher
than the BP of the least volatile cpd.

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GC injection and band broadening

and anomalies.
z Manual injections – takes practice and
patience. Extremely slow injections will cause
band-broadening, wide sample “plug”. Jerky
injections may cause double peaks for the
same analyte species.

z Auto-injectors – robotized carousels of sample

vials. No practice needed. Can be set to make
measurements all night long. Rare in
academe, frequent in industry.

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Split vs. Splitless Injection

z Sample injection is done by a syringe – 1 to 5 µL or ng’s of analyte for
the average capillary column.

z Capillary columns usually require split in injections, a sample reduction

method.

z Depending on the spilt ratio (adjustable) only 0.2 to 2 % of the sample

injection makes its way to the column. The rest is discarded.

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Effect of split versus splitless
injections on separation quality
z The chromatogram
(upper left)
demonstrates ideal
separation
characteristics. Upper
right illustrates what
happens when too
much analyte solute is
injected onto a
capillary column.

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Solid-Phase Microextraction
z Analyte pre-
concentration method,
remember stripping
voltammetry.

z Typical derivatizing
agents:
C18COO-Si-(O)3-

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SPME of Nerve Agents

z Fig 24-21.

Advantages of SPME

“Green” method compared

to solvent extraction

Fibers are reusable 20-30x

102-103 LOD enhancement

Disadvantage – lot’s of
technique and method
development.

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Purge and Trap
Major Goal: Remove all analyte from sample.

Volatile components are removed from

sample by gentle heating and a stream of
purge gas, e.g. He.

Adsorbents trap targeted analytes, e.g.

molecular sieves.

After complete purge of sample, contents of

trap tube are injected into GC.

A purge and trap module is on the Cassini

probe to Saturn and moons looking for
hydrocarbons, H2, He, NH3.
http://saturn.jpl.nasa.gov/home/index.cfm

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GC Detectors
z Ideal detector characteristics, for flowing systems (e.g. GC)

 large dynamic/linear range

 response independent of flow rate, i.e. fast response times.
 Universal detection, responds to all species.

z Keep this in mind when we discuss HPLC and CE.

z Additional requirements for GC

 operation from RT to 400 oC

 detector response independent of detector oven T

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Thermal Conductivity Detector-TCD
z Measures heat loss from a hot filament – nearly universal

z filament heated to const T

 when only carrier gas flows heat loss to metal block is constant, filament T remains
constant

 when an analyte species flows past the filament generally thermal conductivity goes
down, T of filament will rise. (resistance of the filament will rise).

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Flame Ionization Detector (FID).

z Sensitive towards organics

z Analyte is burned in H2/air, which produces CH and CHO+,

radicals, remember our discussion regarding the blue cone in AA.

 CHO+ radicals are reduced at a cathode which produces a current

proportional to the radical quantity. About 10-12 A

 Specific for organic carbon, insensitive to inorganics, CO2, SO2 etc.

 Generally DL 100x less than TCD about pg/s (flow rate dependent)

 Dynamic range of 107

 Response to specific organic depends on the number of organic

carbons.

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Electron Capture Detector (ECD)

z Sensitive to electron withdrawing groups especially towards
organics conatining –F, -Cl, -Br, -I also, -CN, NO2

z Nickel-63 source emits energetic electrons collides with N2

(introduced as make-up gas or can be used as carrier gas)
producing more electrons:

Ni-63 => e-
e- + N2 => 2e- + N2+

z The result is a constant current that is detected by the electron

collector (anode).

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 z As an analyte flows through
past the Ni-63 source
electron capture is possible
by electron-withdrawing
species:

z A + e- => A-

z Current decreases as a
result of e- capture by
analyte. This is one of the
few instances in which a
signal is produced by a
decrease in detectable
phenomenon.

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ECD Characrtersitics (the good)

 VERY low DL for detected species 10-15g/s for many halogenated substances (PCB,
DDT etc).

 OK dynamic range of 104.

z (The bad)

 Radioactive Ni-63 source

 EASILY contaminated with O2, H2O, sample overloading. High maintenance device.

 Highly variable response to halogenated substances, see table below:

 Can be a real headache when method developing a specific analysis, e.g. CH2Cl2 in the
presence of CCl4.

z Sometimes complementary information from FID helps.

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Other GC detectors
z Nitrogen-Phosphorous Detector (NPD)

Also know as the thermionic detector (TID) or alkali flame detector. It

is an FID tweaked for N-P cpds, and organics.

z Flame Photometric Detector (FPD)

FID tweaked for S containing cpds.

z Photoionization Detector (PID)

UV ionization of organic analyte, coupled with high voltage cathode

and analode results in current proportional to ionized organics.

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 z FT-IR we’ve discussed before. (because of generally low ε in A
=εbc, most analytes have a modest DL with FT-IR)

z Mass Spec

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Qualitative Analysis in GC.

z GC-MS offers structural determinations

(remember organic?)

z With other detectors identification is

possible with retention times of analyte
and standard, however it’s best if
another method is used as a
confirmation.

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Quantitative Analysis in GC.

z Calibration curve

z Standard Addition – generally matrix effects

are not as pronounced as in GC as I have
personally noticed in LC.

z Internal Standard – best used with CC and

SA methods. Almost always required since
sample injection is not always a reproducible
phenomenon, even with auto-samplers.

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Method of Internal Standards

z Internal standards are
often used in
chromatographic
methods, but useful in Analyte, X
other techniques where
sample sizes may vary IS, S
from rune to run

z Known quantities of both Signal

non-interfering species
and analyte species are
subjected to the same
analytical procedure and
the signal is measured:

[X] = 0.0837 mM Independent variable

[S] = 0.0666 mM

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F-ratio
The signal of the IS is 347, and that of the analyte is 423.

We now can define a response factor F, defined as

AX A 423 347
=F S = =F
[X ] [S ] 0.0837 0.0666

Solving for F = 0.9700

We should realize that F will not vary even if sample sizes may
do so.

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Example - Now consider an unknown, if 10.0 ml of 0.146 M IS

is added to 10.0 ml of unknown then diluted to 25.0 ml, what is
the unknown analyte concentration is As = 553 and Ax is 582?

[S] = 0.146 M (10.0 ml/25.0 ml) = 0.0584 M

553 582
= 0.9700
[X ] 0.0584

[X] = 0.05721 M

[X]sample = 0.05721 M (25.0 ml/10.0 ml) = 0.143 M

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