Professional Documents
Culture Documents
Chapter 24 - Gas Chromatography
Chapter 24 - Gas Chromatography
Chapter 24 - Gas Chromatography
Chapter 24
Gaseous Mobile Phase, Solid or
Liquid Stationary Phases
1
Carrier
Gas
Reservoir
Gas Flow
Make-up Gas
(optional)
Injection Port
Sample
Introduction
Separation
column
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
2
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.
3
11/22/2005 Chem 253 - Chapter 24 7
4
11/22/2005 Chem 253 - Chapter 24 9
5
“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
6
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
7
Example of the separation efficiency of capillary vs.
packed column.
8
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 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.
Stationary Phase
OH OH OH OH OH OH OH OH
Silica Support
9
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
z Chromatogram 1
= unmodified
z Chromatogram 2
= silinazied
10
Liquid stationary phases
Important Notes:
11
Effect of Stationary Phase Polarity on
Retention Times.
z #1 n-heptane
z #2 tetrahydrofuran
z #3 2-butanone
z #4 n-propanol
12
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).
⎛ α − 1 ⎞⎛ k B ⎞
'
N
Rs = ⎜ ⎟⎜⎜ ⎟
' ⎟
4 ⎝ α ⎠⎝ 1 + k B ⎠
13
Carrier Gases
• O2 is usually avoided since it will oxidize the s.p.
14
Sample Injection
15
GC Injection Syringe
z It is important to rapidly vaporize the sample.
16
11/22/2005 Chem 253 - Chapter 24 33
17
11/22/2005 Chem 253 - Chapter 24 35
18
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.
Solid-Phase Microextraction
z Analyte pre-
concentration method,
remember stripping
voltammetry.
z Typical derivatizing
agents:
C18COO-Si-(O)3-
19
11/22/2005 Chem 253 - Chapter 24 39
Advantages of SPME
Disadvantage – lot’s of
technique and method
development.
20
Purge and Trap
Major Goal: Remove all analyte from sample.
GC Detectors
z Ideal detector characteristics, for flowing systems (e.g. GC)
21
Thermal Conductivity Detector-TCD
z Measures heat loss from a hot filament – nearly universal
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).
Generally DL 100x less than TCD about pg/s (flow rate dependent)
22
11/22/2005 Chem 253 - Chapter 24 45
Ni-63 => e-
e- + N2 => 2e- + N2+
23
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.
z (The bad)
EASILY contaminated with O2, H2O, sample overloading. High maintenance device.
Can be a real headache when method developing a specific analysis, e.g. CH2Cl2 in the
presence of CCl4.
24
11/22/2005 Chem 253 - Chapter 24 49
Other GC detectors
z Nitrogen-Phosphorous Detector (NPD)
25
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
26
Quantitative Analysis in GC.
z Calibration curve
27
F-ratio
The signal of the IS is 347, and that of the analyte is 423.
AX A 423 347
=F S = =F
[X ] [S ] 0.0837 0.0666
We should realize that F will not vary even if sample sizes may
do so.
553 582
= 0.9700
[X ] 0.0584
[X] = 0.05721 M
28