Gas Chromatography

Introduction

1.) Gas Chromatography

Mobile phase (carrier gas) is a gas
- Usually N2, He, Ar and maybe H2
- Mobile phase in liquid chromatography is a liquid

Requires analyte to be either naturally volatile or can be converted to a volatile

derivative
- GC useful in the separation of small organic and inorganic compounds

Stationary phase:
- Gas-liquid partition chromatography nonvolatile liquid bonded to solid support
- Gas-solid chromatography underivatized solid particles
- Bonded phase gas chromatography chemical layer chemically bonded to solid
support

Bonded phase Magnified Pores in activated carbon Zeolite molecular sieve

 Gas Chromatography
Introduction

2.) Instrumentation
Process:
- Volatile liquid or gas injected through septum into heated port
- Sample rapidly evaporates and is pulled through the column with carrier gas
- Column is heated to provide sufficient vapor pressure to elute analytes
- Separated analytes flow through a heated detector for observation
 Gas Chromatography
Instrumentation

1.) Open Tubular Columns

Commonly used in GC
Higher resolution, shorter analysis time, and greater sensitivity
Low sample capacity

Increasing Resolution
- Narrow columns Increase resolution

- Resolution is proportional to N , where N increases directly with column length

Easy to generate long (10s of meters)

lengths of narrow columns to maximize
resolution
 Gas Chromatography
Instrumentation

1.) Open Tubular Columns

Increasing Resolution

Decrease tube diameter

Increase resolution
Increase Column Length
Increase resolution
 Gas Chromatography
Instrumentation

1.) Open Tubular Columns

Increasing Resolution

Increase Stationary Phase Thickness

Increase resolution of early eluting compounds

Also, increase in
capacity factor and
reduce peak tailing

But also decreases

stability of stationary
phase
 Gas Chromatography
Instrumentation

2.) Choice of liquid stationary

phase:
Based on like dissolves
like

Nonpolar columns for

nonpolar solutes

Strongly polar columns for

strongly polar compounds

To reduce bleeding of
stationary phase:
- bond (covalently
attached) to silica

- Covalently cross-link to
itself
 Gas Chromatography
Instrumentation

3.) Packed Columns

Greater sample capacity
Broader peaks, longer retention times and less resolution
- Improve resolution by using small, uniform particle sizes

Open tubular column

Packed column
 Gas Chromatography
Instrumentation
500 L chromatography
3.) Packed Columns column
The major advantage and use is for large-scale or
preparative purification

Industrial scale purification maybe in the kilogram or

greater range

Oil refinery separates

fractions of oil for
petroleum products
 Gas Chromatography
Retention Index

1.) Retention Time

Order of elution is mainly determined by volatility
- Least volatile = most retained
- Polar compounds (ex: alcohols) are the least volatile and will be the most
retained on the GC system

Second factor is similarity in polarity between compound and stationary

phase
 Gas Chromatography
Retention Index

2.) Describing Column Performance

Can manipulate or adjust retention time by changing polarity of stationary
phase

Can use these retention time differences to classify or rate column

performance
- Compare relative retention times between compounds and how they
change between columns
Can be used to identify unknowns
 Gas Chromatography
Retention Index

2.) Describing Column Performance

Retention index based on the difference in the number of carbons (N, n) for
linear alkane and corresponding retention times (tr(unknown), tr(N),tr(N)):
log t'r ( unknown ) log t'r ( n )
I 100 n ( N n )
log t'r ( N ) log t'r ( n )

Provides a means to compare the performance of different columns

Increase in Polarity
 Gas Chromatography
Temperature and Pressure Programming

1.) Improving Column Efficiency

Temperature programming:
- Temperature is raised
during the separation
(gradient)

- increases solute vapor

pressure and decrease
retention time

Temperature gradient improves

resolution while also decreasing
retention time
 Gas Chromatography
Temperature and Pressure Programming

1.) Improving Column Efficiency

Pressure Programming:
- Increase pressure increases flow of mobile phase (carrier gas)
- Increase flow decrease retention time

Van Deemter curves indicate

that column efficiency is
related to flow rate

Pressure is rapidly reduced at the end of the run

- Time is not wasted waiting for the column to cool
- Useful for analytes that decompose at high temperatures
 Gas Chromatography
Carrier Gas

1.) N2, He and H2 are typical carrier gases

He:
- Most common and compatible with most
detectors
- Better resolution (smaller plate heights)
- Solutes diffuse rapidly smaller mass
transfer term
N2:
- Lower detection limit for a flame
ionization detector
- Lower resolution and solute diffusion
rates
H2:
- Fastest separations
- Can catalytically react with unsaturated
compounds on metal surfaces
- Can not be used with mass spectrometers Flow rate increases N2 < He < H2
Forms explosive mixtures with air
- Better resolution (smaller plate heights)
- Solutes diffuse rapidly smaller mass Diffusion coefficients follow: H2 > He > N2
transfer term
Gas Chromatography
Sample Injection

1.) Sandwich Injection

Separate sample with air bubbles and solvent

- Air bubble prevents depletion of most volatile compounds before sample

injection is complete (barrier between oven and sample during injection)

- Solvent is used to pushes out sample, but bubble prevents mixing

- Final air bubble pushes out solvent

- Gas-tight syringe is required for gas samples

- Injection volume is typically 0.1-2 mL

Gas Chromatography
Sample Injection

1.) Sandwich Injection

Injection port

- Inject rapidly ( < 1s) through septum into evaporation zone

- Injector temperature is kept high (350oC) for fast evaporation

- Rapid gas flows carries sample to mixing chamber for complete vaporization
and complete mixing before entering column
 Gas Chromatography
Sample Injection

2.) Split Injection

Delivers only 0.2-2% of sample to the column
- Split ratio of 50:1 to 600:1 (sample discarded)
For samples where analytes of interest are >0.1% of sample
- Best resolution is obtained with smaller amount of sample
- 1 mL with 1 ng of each compound (0.5 mL of gas volume)
Not quantitative, split not constant

Remainder of the sample is

flushed from injector port to After mixing, pressure
column regulator controls the
fraction of sample discarded
Gas Chromatography
Sample Injection

2.) Splitless Injection

Delivers ~80% of sample to the column
For trace analysis, where analytes of interest are < 0.01% of sample
- Large volume (~2 mL) injected slowly (2s)
No mixing chamber or split vent
- Injection temperature is lower (220oC)
- 40oC below the boiling point of the solvent

Injecting larger volume, dont

want broad peaks

Lower temperature traps

solvent in a narrow band at
the head of the column

Raise temperature to volatize

sample and start separation
Gas Chromatography
Sample Injection

2.) Splitless Injection

Solvent trapping significantly improves the performance of splitless
injections
- Initial lower temperature of column during injection keeps larger volume into
a narrow band
- Chromatography is initiated by raising column temperature
- Cold trapping condense solutes in narrow band at the beginning of column
by using an initial temperature 150oC below boiling points of solutes of
interest

With Solvent trapping

Without Solvent trapping
Gas Chromatography
Sample Injection

3.) On-column Injection

Delivers ~100% of sample to the column
For samples that decompose above their boiling points
Solution injected directly on column
- Warming column initiates chromatography

Lower initial column temperature

to prevent sample decomposition

Raise temperature to volatize

sample and start separation
Gas Chromatography
Detectors

1.) Qualitative and Quantitative Analysis

Mass Spectrometer and Fourier Transform Infrared Spectrometers can
identify compounds as part of a GC system
- Compare spectrum with library of spectra using a computer

Compare retention times between reference sample and unknown

- Use multiple columns with different stationary phases
- Co-elute the known and unknown and measure changes in peak area

The area of a peak is proportional to the quantity of that compound

Area of Gaussian peak 1.064 peak height w1
2

Peak area increases proportional

Peak Area

to concentration of standard if
unknown/standard have the
identical retention time same
compound

Concentration of Standard
Gas Chromatography
Detectors

2.) Thermal Conductivity Detector

Measures amount of compound leaving column by its ability to remove heat
- He has high thermal conductivity, so the presence of any compound will
lower the thermal conductivity increasing temperature of filament
As heat is removed from filament, the resistance (R) of filament changes
- Causes a change in an electrical signal that can be measured
Responds to all compounds (universal)
- Signal changes in response to flow rate of mobile phase and any impurities
present
- Not very sensitive

Ohms Law: V =IR

Based on Ohms law, monitored

potential (V) or current (I) Changes
as resistance (R) of filament changes
due to presence of compound
Gas Chromatography
Detectors

3.) Flame Ionization Detector

Mobile phase leaving the column is mixed with H2 and air and burned in a flame
- Carbon present in eluting solutes produces CH radicals which produce CHO+ ions
- Electrons produced are collected at an electrode and measured
Responds to almost all organic compounds and has good limits of detection
- 100 times better than thermal conductivity detector
- Stable to changes in flow rate and common mobile phase impurities (O2, CO2,H2O,NH3)

Burn sample and measure

amount of produced electrons
Gas Chromatography
Detectors

4.) Electron Capture Detector

Sensitive to halogen-containing and other electronegative compounds
Based on the capture of electrons by electronegative atoms
- Compounds ionized by b-rays from radioactive 63Ni
Extremely sensitive (~ 5 fg/s)

Steady current (flow of electrons)

disrupted by compounds with
high electron affinity
Gas Chromatography
Detectors

5.) Mass Spectrometry

Detector of Choice But Expensive!
Sensitive and provides an approach to identify analytes
Selected ion monitoring monitor a specific mass/charge (mz) compared to
scanning over the complete spectra
- Simplifies complex chromatogram
- Increases sensitivity by 102-103
Gas Chromatography
Detectors

6.) Other Detectors

Respond to limited class of analytes
Modification of previous detectors
Nitrogen-Phosphorous detector
- Modified flame ionization detector
- Extremely sensitive for compounds containing N and P
- Important for drugs and pesticides

Flame photometric detector

- Measures optical emission from P (536 nM) , S (394 nM), Pb, Sn, and other
select elements after passing sample through flame (flame ioniation
detector)

Photionization detector
- Uses a ultraviolet source to ionize aromatic and unsaturated compounds,
electrons produced are measured (Electron capture detector)

Sulfur/nitrogen chemiluminescence detector

- Collects exhaust of flame ionization detector
- S and N converted to SO and NO
- Mix with O3 form excited state of SO2 (emits blue light) and NO3
Gas Chromatography

Sample Preparation

1.) Transform sample into form suitable for analysis

Extraction, concentration, removal of interfering species or chemically
transforming (derivatizing)

2.) Solid-phase microextraction

Extract analytes from complex
mixture without solvent
Uses a fused-silica fiber coated
with stationary phase
- Stationary phase similar to
those used in GC
Expose Fiber to sample to
extract compounds and then
inject fiber into GC to
evaporate analytes
 Gas Chromatography
Sample Preparation

3.) Purge and Trap

Removes volatile analytes from liquids or solids,
concentrates sample and transfer to GC

Goal is to remove 100% of analyte

Connect port to GC

Heat column to
Analytes are captured
200oC to transfer
on adsorbent column
analytes to GC

Bubble purge gas (He)

through heated sample to
evaporate analytes
Gas Chromatography
Method Development in GC

1.) How to Choose a Procedure for a Particular Problem

Many Satisfactory Solutions
The order in which the decision should be made should consider:
1. Goal of the analysis
2. Sample preparation
3. Detector
4. Column
5. Injection

Goal of the analysis

- Qualitative vs. quantitative
- Resolution vs. sensitivity
- Precision vs. time
- Interest in a specific analyte

Sample preparation
- Cleaning-up a complex sample is essential
- Garbage in garbage out

Choosing the Detector

- Detect a specific analyte(s) or everything in the sample
- sensitivity
- Identify an unknown (MS, FTIR)
Gas Chromatography
Method Development in GC

1.) How to Choose a Procedure for a Particular Problem

Selecting the Column
- Consider stationary phase, column diameter and length, stationary phase
thickness
- Match column polarity to sample polarity
- To improve resolution, use a:
a. Longer column
b. Narrower column
c. Different stationary phase

Choosing the Injection Method

- Split injection is best for high concentrated samples
- Splitless injection is best for very dilute solutions
- On-column injection is best for quantitative analysis and thermally instable
compounds