Dong-Sun Lee/ CAT-Lab / SWU

2012-Fall version

Chapter 31

Gas Chromatography (GC)

Introduction Gas chromatography is a chromatographic technique that can be used to separate volatile organic compounds. Two types of GC are encountered: gas-solid chromatography(GSC) and gas-liquid chromatography(GLC). GLC is finds widespread use in all fields of science, where its name is usually shortened to GC. A gas chromatograph consists of a flowing mobile phase, an injection port, a separation column containing the stationary phase, and a detector. GSC is based on a solid stationary phase on which retention of analytes is the consequence of physical adsorption. GLC is based on partitioning behavior of the analyte between the mobile gas phase and the liquid stationary phase in the column.

Characteristics of GLC 1. Sensitivity : mg ~ pg (103 ~ 109 g) 2. Versatility : from rare gases to liquids and solids in solution with 800~1000 MW 3. Speed of analysis : typically 5 ~ 30 min , complex 3 ~ 30 mixture separation 4. Reproducibility : qualitative Accuracy : quantitative

HP 5890 gas chromatograph (Hewlett Packard) with an integrator

GC-MS : GC-Q plus ion-trap GC-MSn ( Thermoquest - Finnigan ) Xcalibur software

GC-14A gas chromatograph (Shimadzu) with an integrator.

Schematic diagram of a gas chromatograph system. Valve/ Gage

Fuel

Trap

Make-up gas Injector

gas Detector Electrometer

Regulator

Split vent

Column Column oven

Recorder Integrator Computer

Fuel
gas

Carrier
gas Basic components of Gas Chromatograph Carrier gas supply Sample introduction inlet Column and controlled-temperature oven Detector & oven Recorder

Carrier gas system in gas chromatograph

The purpose of the carrier is to transport the sample through the column to the detector. The selection of the proper carrier gas is very important because it affects both column and detector performance. The detector that is employed usually dictates the carrier to be used. From a column performance point of view a gas having a small diffusion coefficient is desirable (high molecular weight, e.g., N2, CO2, Ar) for low carrier velocities while large diffusion coefficients (low molecular weight, e.g., H2, He) are best at high carrier velocities. The viscosity dictates the driving pressure. For high-speed analysis, the ratio of viscosity to diffusion coefficient should be as small as possible. H2 would be the best choice, followed by helium. The purity of the carrier should be at least 99.995% for best results. Impurities such as air or water can cause sample decomposition and column and detector deterioration. In temperature programmed runs, impurities in the carrier gas such as water can be retained at low temperatures but are then eluted at higher temperatures impairing the baseline. Many instrument problems have been traced to contaminated carrier gases. The carrier must also be inert to the components of the sample and the column.

Properties of common carrier gases

Gas molecular weight Thermal conductivity 105 at 100oC (g-cal/sec-cm- oC ) Viscosity 106 at 100oC (P)

Ar

39.95

5.087

270.2 *

CO2
He H2 N2 O2 * at 99.6oC

44.01
4.00 2.016 28.01 32.00

5.06
39.85 49.94 7.18 7.427

197.2
234.1 104.6** 212.0 248.5***

** at 100.5oC

**** at 99.74oC

Using the correct carrier and detector gases are an important factor in installing a new GC. The five gases commonly used as carrier gas and detector fuels in capillary gas chromatography are helium, hydrogen, nitrogen, argon-methane, and air. The types of gases necessary are partly determined by the detection system used. Factors to consider for each individual gas are discussed below.

Carrier Gas Choice Carrier gases that exhibit a broad minimum on a van Deemter profile are essential in obtaining optimum performance. Volumetric flow through a capillary column is affected by temperature. When temperature programming from ambient to 300oC, the flow rate can decrease by 40 percent. A carrier gas that retains high efficiency over a wide range of flow rates and temperatures is essential in obtaining good resolution throughout a temperature programmed run. Figure 1 shows the van Deemter profile for hydrogen, helium, and nitrogen carrier gases.

Van Deemter curves for GC of n-C17H36 at 175oC, using N2, He, or H2 in a 0.25 mm diameter 25 m long wall coated column with OV-101 stationary phase

Hydrogen is the fastest carrier gas (uopt), with an optimum linear velocity of 40cm/sec, and exhibits the flattest van Deemter profile. Helium is the next best choice, with an optimum linear velocity of uopt = 20cm/sec. Nitrogen's performance is inferior with capillary columns because of its slow linear velocity, uopt = 12cm/sec. Argon-methane has a slower optimum linear velocity than nitrogen and is not recommended for use as a carrier gas with capillary columns. Air is not recommended as a carrier gas because it can cause stationary phase oxidation. With hydrogen and helium as carrier gases, the minimum H.E.T.P. values can be maintained over a broader range of linear velocities than with nitrogen, and high linear velocities can be used without sacrificing efficiency. Nitrogen is beneficial only when analyzing highly volatile gases under narrow temperature ranges where increasing stationary phase interaction is desirable. Otherwise, the use of N2 results in longer analysis times and a loss of resolution for compounds analyzed on a wide temperature range.
http://www.restekcorp.com/gcsetup/gcsetup3.htm

Exert Caution when Using Hydrogen as a Carrier Gas

Hydrogen is explosive when concentrations exceed 4% in air. Proper safety precautions should be utilized to prevent an explosion within the column oven. Most gas chromatographs are designed with spring loaded doors, perforated or corrugated metal column ovens, and back pressure/flow controlled pneumatics to minimize the hazards when using hydrogen carrier gas. Additional precautions include: Frequently checking for leaks using an electronic leak detector. Using electronic sensors that shut down the carrier gas flow in the event of pressure loss. Minimizing the amount of carrier gas that could be expelled in the column oven if a leak were to occur by installing a flow controller (needle valve) prior to the carrier inlet bulkhead fitting to throttle the flow of gas (for head pressure controlled systems only) as shown Fig. 2. Fully open the flow controller (needle valve) and obtain the proper column head pressure, split vent flow, and septum purge flow rates. Decrease the needle valve flow rate until the head pressure gauge begins to drop (throttle point). Next, increase the flow controller (needle valve) setting so that the right amount of flow is available to the system. Should a leak occur, the flow controller will throttle the flow, preventing a large amount of hydrogen from entering the oven.

Make-up and Detector Fuel Gases

Gas added to the stream after the column is called makeup gas. Choosing the correct make-up and detector gases will depend on both the detector and application. Most GC detectors operate best with a total gas flow of approximately 30ml/min. to ensure high sensitivity and excellent peak symmetry. Refer to your GC manual for optimum flow rates on different instruments. Carrier gas flows for capillary columns range from 0.5 to 10ml/min. which are well below the range where most detectors exhibit optimal performance. To minimize detector dead volume, make-up gas is often added at the exit end of the column to increase the total flow entering the detector. Make-up gas helps to efficiently sweep detector dead volume thereby enhancing detector sensitivity.
Make-up gas can be added directly to the hydrogen flame gas for flame ionization detectors (FID), nitrogen phosphorous detectors (NPD), and flame photometric detectors (FPD) or added to the column effluent by an adaptor fitting. However, GCs such as Perkin-Elmer and Fisons do not require make-up gas. Combustion type detectors (FID, NPD, FPD) use three gases: make-up, hydrogen (fuel gas), and air (combustion/oxidizing gas). For non-combustion detectors, such as the thermal conductivity detector (TCD), electron capture (ECD), and photo ionization detector (PID), only carrier and make-up gases are required. In the case of the electrolytic conductivity detector (ELCD), the make-up gas is hydrogen, as a reaction gas in the halogen and nitrogen mode or air in the sulfur mode. Table I shows recommended gases for various detectors.

Carrier gases and detector fuel gases for use with various GC detectors
TCD Carrier Gases He H2 N2 Combustion/ Reaction Gases H2 O O O ECD O O FID O O O O NPD O O O FPD O O O O ELCD O O O PID O O O -

Air
Make-up Gases N2 He ArCH2

O O -

O O

O
O O -

O
O O -

O
O O -

O O -

http://www.restekcorp.com/gcsetup/gcsetup3.htm

Effect of impurities
- Impurities such as hydrocarbon, oxygen, water contribute to unwanted noise levels, excessive baseline drift. - Molecular sieve --- Moisture trap, Oxygen trap, Chemical filter

Effect of water on column efficiency - Carrier gas dryness is very important !! (use anhydrous sodium sulfate) - Water can and usually does react with some portion of the column. This results in loss of resolution and tends to produce asymmetric or tailing peaks. Unwanted components or ghost peaks may also appear. Another effect is a net loss of sensitivity.

Gas purifiers
The trap will remove any water vapor or oils that may have been introduced in the filling process since a number of gases are water pumped. The contaminants removed by the trap could otherwise interact with the column packing material to produce spurious peaks. In addition the contaminants can cause increased detector noise and drift.

The traps should be reconditioned (about twice a year ) by heating to 300 oC for 4~8 hr with a stream of gas passing through it or in a vacuum oven.

Carrier gas purifiers

1
Gas cylinder

GC

1. Hydrocarbon trap 2. Moisture trap 3. Oxygen trap

4. Indicating oxygen trap

Gas purifier recommendation for GC applications

Capillary column GC with any detector Carrier Make-up Hydrocarbon, Moisture, Oxygen None - all detector but ECD moisture & oxygen Air for FID H2 for FID Hydrocarbon None

ELCD reaction gas Hydrocarbon Packed column GC with FID or TCD Carrier Hydrocarbon, Moisture, Oxygen

Packed column GC with ECD, FPD, NPD, MSD

Carrier

Hydrocarbon, Moisture, Oxygen

Flow requirements 1. Stable 2. Reproducible 3. Convenient The more constant the flow rates, the more precise and accurate the results. Flow controller ( Pressure controller ) --- to maintain precise and accurate flow rates

Effect of decreased flow rate or lower temperature

- All peaks have shifted to longer retention times - Apparent loss of peak height - The base of each peak is wider, however, individual peak area remain constant.

Effect of increased flow rate

- Sample components are squeezed toward the injection point - Cause two components to elute together, appearing as single peak

Regulations of carrier gas

Carrier cylinder bottled at about 2500 psi(150-160 atm)

Two stage pressure regulator : - first stage : high inlet pressure

- second stage : low outlet pressure

( set at 40~100psi)

Gas generators

Gas flow rate control

A 1 % change in carrier gas flow rate will cause a 1% change in retention time. For all these reasons it is important to keep the flow of the carrier gas constant.
1. Control of carrier gas inlet pressure 2. Control of carrier gas flow rate In isothermal operation the means of regulation is immaterial because both means provide constant inlet pressure as well as constant flow rate. In temperature programmed runs, however, the situation is quite different. If one maintains the inlet pressure constant the flow rate will change. Therefore, with temperature programming of the column, the flow rate must be controlled.

Pressure controllers 1. The second stage regulator on the cylinder 2. A pressure regulator mounted in the GC 3. A needle valve(variable restrictor) mounted in the GC 4. A fixed restrictor mounted in the GC

Flow measurement
1) Rotometer

The column flow rate is typically indicated by a rotometer . ( Calibrate equilibrium position indicating the flow )
Rotometer is operated by the volume of gas passing a ball in a tapered cell. 2) Bubble meter 3) Electronic flow sensor

Relationships between inside diameter, column length, mesh size, and carrier gas flow for packed column

Inside diameter
mm

Mesh size for

length up to 3m

Mesh size for

length over 3m

Carrier flow
N2, ml/min He or H2, ml/min

2
3 4

100~120
100~120 80~100

80~100
80~100 60~80

8~15
15~30 30~60

15~30
30~60 60~100

John A. Dean, Analytical Chemistry Handbook, McGraw-Hill, 1995, p. 4.31 .

Comparison of 1/8-in(0.316 cm) packed, wide bore, and WCOT columns

1/8 in packed Inside diameter, mm Film thickness, m Phase volume ratio() Column length, m Flow rate, ml/min Effective plates(Neff) per meter Effective plate height (Heff),mm Typical sample size 2.2 5 15~30 1~2 20 2000 0.5

Wide bore 0.53 1~5 130~250 15~30 5 1200 0.6 15 g

WCOT 0.25 0.25 250 15~60 1 3000 0.3 50 ng

John A. Dean, Analytical Chemistry Handbook, McGraw-Hill, 1995, p. 4.31 .

Recommended ranges of gas flow rates

Detector FID Gases Carrier Hydrogen Air Make-up(N2) NPD Hydrogen 30~50 ml/min 300~600 ml/min 10~60 ml/min 2~4 ml/min Range Capillary 2 ml/min 35 ml/min 350 ml/min 30 ml/min Packed 40 ml/min 40 ml/min 500 ml/min not used

Air

40~80 ml/min
not necessary

Make-up(N2, He) 10~20 ml/min PID Make-up Sheath FPD Carrier Hydrogen Air 5~10 ml/min 30~40 ml/min 1~3 ml/min 85~100 ml/min 100~120 ml/min

30~50 ml/min 100~120 ml/min 110~135 ml/min

Inlet requirements
1. Temperature controlled 2. Low volume (total swept by carrier ) 3. Inert construction

Column Overload
If too large an sample were allowed to enter small bore(capillary) columns column overload and a loss of resolving power would like occur.

Liner
All liners help protect the vaporized sample form contacting the metal wall of the inlet as sample flows onto the column. Deactivated glass wool may be used as an aid for sample vaporization, to minimize discrimination based on boiling point, and to provide a surface on which non0volatiles can be trapped. The simplest liner is a straight tube, which gives allaround good performance at low cost. Single-taper liners improve on a straight tube by minimizing sample vapor contact with metal at the bottom of the injection port, although they are somewhat more expensive. Liners are deactivated borosilicate glass, except quartz where noted. Liners are guaranteed inert for phenols, organic acids and bases.

Why is Glass Wool Added to an Injector Liner? - General GC

The Glass wool serves three major purposes. The Glass wool will prevent the small pieces of septa from reaching the column. The presence of Glass wool will help the injected sample stay in the liner a little longer which will help the sample to vaporize and mix more thoroughly with the carrier gas. If positioned properly it will wipe the outer surface of the syringe needle and improve the precision of the liquid injection.

Inlet configuration
1. Direct column inlet --- 1/8 " OD or larger column

sampling syringe is actually inserted into the end of the column needle guide / cap / spring / septum mounting holes / carrier gas in / inlet body column Swagelock ferrules / Swagelock nut 2. Splitter inlet --- open tubular column or less than 1/8" OD column
Because of the limited capacity for sample of these small bore columns and the difficulty of injecting extremely small volume samples, a large portion of the injected sample is vented to atmosphere by the inlet. septum / preheated carrier gas / mixing tube / restrictor buffer volume / tapered needle / gold gasket / column fitting

Injection port for split injection into an open tubular column. The glass liner is slowly contaminated by nonvolatile and decomposed samples and must be replaced periodically. For splitless injection, the glass liner is a atraight tube with no mixing chamber. For dirty samples, split injection is used and a packing material can be replaced inside the liner to adsorb undesirable components of the sample.

Common injection techniques

1) Hot flash vaporization
Direct Cold-trap Split Splitless 2) Direct cold : on column

Split or on-column
Split 1) Simple 2) High column efficiency 3) Column may be protected On-column 1) Best accuracy 2) Thermolabile compounds 3) Trace analysis

Representative injection conditions for split, splitless, and on-column injection into an open tubular column.

Direct injector
1) Good sensitivity 2) Low column efficiency 3) Best for thick films, widebore column ( 0.53 mm )

Hot on-column injectors

1) Reduced column efficiency 2) Best with thick films, widebore columns 3) Nonvolatiles may damage column 4) Cold on-column injector may be used with 0.1 to 0.53 mm i.d. columns

Advantage of on-column
1) Best reproducibility : Quantitative results 2) No split, no loss of high boilers 3) "Cold" on-column injection available

Advantage of splitless
1) High sensitivity ( 95 % of sample on column ) 2) Solvent effect produces narrow sample bands 3) Same hardware as split injection

Disadvantage of splitless
1) Slow sample transfer to column 2) Must dilute sample with volatile solvent 3) Time consuming : must cool column 4) Poor for thermolabile compounds

Split and splitless injections of a solution containing 1 vol % methyl isobutyl ketone (bp 118 oC) and 1 vol % p-xylene (bp 138 oC) in dichloromethane (bp 40 oC) on a BP10 moderately polar cyanopropyl phenyl ,ethyl silicone open tubular column(0.22 mm I.d., 10 m long, 0.25 m, column temperature =75 oC).

Common injection methods

Syringe injection
Valve injection

Sampling Syringe
0 ~ 1 L --- the sample is totally confined to the needle 0 ~ 5 / 0 ~ 10 L needle / barrel / plunger

Gas sampling valve

Sample in Sample loop --- 1/4 or 10 mL loop size compatible with needs Sample vent Carrier gas to column

Solvent effect
t-1 t-2

time t-1 --- just after injection, solvent and sample are condensed in a long plug at the front of the column. The column temperature must be cold enough to condense the solvent. time t-2 --- after some time, the column temperature has been raised, most of the solvent has evaporated, and the solvent effect has left the sample molecules concentrated in a narrow band. As the column is further heated, the remaining solvent and sample molecules are rapidly vaporized --- resulting in high column efficiency and narrow peak.

Syringe for solid phase microextraction. Sampling by SPME and desorption of analyte from the coated fiber into a gas chromatograph.

Purge and trap apparatus for extracting volatile substances from a liquid or solid by flowing gas.

GC column
Parts of Column
1) Tubing material Stainless steel--- reactive ( steroids, amines, free acids ) Glass ------------ can be made inert, difficult handling Fused silica ---- flexible most inert most popular high resolution 2) Stationary phase Solid support --- carefully sized granular

Liquid phase --- active portion of the column

Porous polymers Adsorbents

Important column parameters 1) Inside diameter 2) Length 3) Film thickness 4) Stationary phase composition 5) Flow rate

Column diameter
i.d. Resolution Speed Capacity Ease

100 micrometer (narrow bore ) 250, 320 (mid bore) 530 (wide bore)
Column length N L

+++
++

+++
++

+
++

+
++

++

+++

+++

R L1/2

tR L

Column
24-foot 1/8" packed column wound on 6" coil 6-foot 1/4" packed column wound on 5" coil 60-meter 0.53mm metal wide bore capillary column wound on 3.5" coil 15-meter 0.53mm fused silica wide bore capillary column wound on 7" cage
Glass wool --- both ends of the column 1- (inlet side) 1/4 (detector side)

30-meter .25mm metal narrow bore capillary column wound on 3.5" coil
http://www.srigc.com/catalog/columns.htm

Fused silica surface

made by the reaction of SiCl4 and water vapor in a flame - SiO2 contains 0.1 % OH groups - Very inert - Uniform chemical surface

Fused silica
- High tensile strength - Flexible - Sheath of polyimide - Very inert

Fused Silica Capillary Columns

A fused silica capillary column is comprised of three major parts (Figure 1). Polyimide is used to coat the exterior of fused silica tubing. The polyimide protects the fused silica tubing from breakage and imparts the amber-brown color of columns. The stationary phase is a polymer that is evenly coated onto the inner wall of the tubing. The predominant stationary phases are silicon based polymers (polysiloxanes), polyethylene glycols (PEG, Carbowax) and solid adsorbents. Figure 1. Capillary columns have to be properly installed to maximize their performance and lifetime. You can obtain enhanced column performance and lifetime by following these recommended installation guidelines. More detailed installation, operational and troubleshooting information can be found in the following references

WCOT = Wall Coated Open Tubular

invented and patented by Dr Marcel Golay

Tubing - Fused silica - Glass - Stainless steel Liquid phase coating WCOT - - - High resolution
Film thickness : 0.5 to 5.0 micrometer i.d. : 0.10, 0.25, 0.32, 0.53 mm Length : 10 to 60 m

Open tubular GC column

Operational guideline for open tubular GC columns

WCOT
narrow Column inner diameter, mm Maximum sample volume, l Maximum amount for one component, ng Effective plates(Neff) per meter 2~50 3000~5000 3~75 2500~4000 5~100 1500~2500 0.25 0.5 intermediate 0.32 1 wide bore 0.53 1

Trennzahl(separation) number
per 25 m Optimum flow for N2, ml/min * Optimum flow for He, ml/min ** Optimum flow for H2, ml/min *** 40 0.5~1 1~2 2~4 35 0.8~1.5 1~2.5 3~7 25 2~4 5~10 8~15

* Optimum velocity is 10 to 15 cm/s for each column ** Optimum velocity is 25 cm/s for each column *** Optimum velocity is 35 cm/s for each column

Other types of capillary columns

SCOT = Support Coated Open Tubular Solid support : Celite Liquid phase Not available fused silica tubing PLOT = Porous Layer Open Tubular Porous adsorbent : alumina or molecular sieve * Molecular sieve --- efficient for H2, Ne, Ar, O2, N2, CO, CH4.

Porous carbon stationary phase ( 2 m thick) on inside wall of fused silica open tubular column.

Capillary column vs Packed column

Capillary Length 60 m Packed 2m

Theoretical plates(N/m)
Total plates length(N/m)

3000-5000
180000-300000

2000
4000

Advantages of capillary column

1) Not packed : long lengths : high resolution 2) Thin film ; efficient, fast 3) Used silica --- inert surface, better results

Disadvantages of capillary column

1) More expensive 2) Limited liquid phase Requires very small samples 3) Dedicated instrumentation --capillary inlets, septum purge

(Left) Gas chromatogram of alcohol mixture at 40 oC using packed column ( 2mm I.D., 76 cm long containing 20 % Carbowax 20 M on a Gas-Chrom R support and FID. (Right) Chromatogram of vapors from headspace of beer can, obtained with 0.25 mm diameter, 30 m long porous carbon column oerated at 30 oC for 2 min and then ramped up to 160 oC at 20 oC/min.

Component Separation with the Column < The process of separation >
A series of partitions : Dynamic In-and Out (or Stop-and-Go) All differential migration process. The most volatile components usually pass through the column first, the least volatile or highest boiling emerges last.

Analytes

Mobile phase( Driving force)

Stationary phase ( Resistive force)

Capillary Column Installation Steps

1. Check traps, carrier gas, septum, liner 2. Place the nut and ferrule on the column and carefully cut the column end 3. Install the column into the injector 4. Turn on the carrier gas 5. Install the column into the detector 6. Inspect for leaks 7. Confirm carrier gas flow and proper column installation 8. Condition the column 9. Accurately set the carrier gas velocity 10. Bleed test 11. Run test mix
Click here for a complete listing of tools available from J&W, including magnifiers and cutting tools. http://www.jandw.com/gccolumn.htm#Fused Silica Capillary Columns

Recommended Installation Tools and Supplies

1. Cutting tool such as a diamond or carbide tipped pencil, sapphire tipped pencil or ceramic cleaving wedge 2. Magnifier (10-20X) 3. Ruler 4. Wrench 5. Ferrules 6. Vial of solvent 7. Clean syringe 8. Supply of an appropriate non-retained compound 9. Column test mixture 10. Flow meter 11. Other supplies: septa, clean injector liners, liner ferrules/O-rings, etc.

Conditioning of the Column Once the column has been checked for proper installation and the absence of leaks, it is ready for conditioning. Heat the column to its isothermal, upper temperature limit (temperature limits listed below) or a temperature 10-20 oC above the highest operating temperature of your particular method. Do not exceed the upper limit or column damage will result. Heat the column rapidly - slow temperature programming is not necessary. After the column has reached the conditioning temperature, plot the baseline. Keep the baseline on scale so that it can be observed. The baseline should be elevated at first then start to drop after 510 minutes at the conditioning temperature. The baseline will continue to drop for 30-90 minutes then stabilize at a constant value. If the baseline does not stabilize after 2-3 hours or does not start to significantly decrease after 15-20 minutes, either a leak is present or a contamination problem exists. In either case, immediately cool the oven down below 40 oC and resolve the problem. Continued conditioning will result in column damage or the inability to obtain a stable baseline. Excessive conditioning of the column may result in a shortened lifetime. In general, polar stationary phases and thick film columns usually require longer times to stabilize than less polar and thinner film columns. GS PLOT columns require a different conditioning procedure than liquid stationary phase columns.

Conditioning Procedure for GS-Q PLOT Columns

GS-Molesieve, GS-Alumina and GS-Q columns require special conditioning procedures. The following conditions are recommended. Column: Temperature: GS-Q: 250 oC for a minimum of 8 hours GS-Molesieve: 300 oC for 3 hours or 250 oC for 12 hours GS-Alumina: 200 oC for a minimum of 8 hours GS-Alumina and GS-Molesieve are susceptible to retention shifts from reversible absorption of water. If retention shifts are observed after analyzing high water content samples, recondition the column to remove any water trapped by the stationary phase. If the column is conditioned with the detector end disconnected, a small portion of the columns end may be damaged. Remove 10-20 cm of the exposed column end before installing the column into the detector.

Temperature Limits
The temperature limits define the range over which the column can be safely used. If the oven is operated below the lower temperature limit, poor separation and peak shape problems will be evident, but no column damage will occur. Upper temperature limits are usually given as two numbers. The first or lower temperature of the two is the isothermal limit. The column can be maintained at this temperature for indefinite periods of time. The second or higher temperature is the program limit. The column can be heated to this temperature for short periods of time (<10 minutes). Exceeding the upper temperature limits will significantly reduce column lifetime.

Liquid Phase(=Stationary phase) Classes

1. Non-polar phase gives boiling point order separation 2. Selective phase separates components that have close b.p. and small structural differences 3. Polar phase depends on internal functional groups to separate compounds that have reactive OH, NH2 or other polar radicals 4. Each stationary phase retains solutes in its own class best

Raising percentage of stationary phase leads to

1) Greater capacity for solute 2) Longer retention time 3) Increased HETP

Column Dimethylpolysiloxane

Temperature Polyethylene Glycol

DB-1; 0.1, 0.25 and 1.0 m

DB-1; 3.0 and 5.0 m DB-1; Megabore 0.1 m DB-1; Megabore 1.5 m DB-1; Megabore 3.0 and 5.0 m DB-1ht (5%-Phenyl) Methylpolysiloxane DB-5 DB-5; Megabore DB-5; Megabore 5.0 m DB-5ht DB-5ms DB-5ms; Megabore DB-5.625

-60 to 325/350 oC
-60 to 280/300 oC -60 to 360 oC -60 to 300/320 oC -60 to 260/280 oC -60 to 400 oC

DB-WAX DB-WAX; Megabore Base Deactivated Polyethylene Glycol CAM Carbowax 20M Carbowax

20 to 250/260 oC 20 to 230/240 oC

60 to 220/240 oC

60 to 220/240 oC

-60 to 325/350 oC -60 to 300/320 oC -60 to 260/280 oC -60 to 400 oC -60 to 325/350 oC -60 to 300/320 oC -60 to 325/350 oC

-Cyclodextrin for chiral column.

Column oven and temperature control

Oven size : column sits in an oven Inner volume : laboratory GC ; about 22-25 L process GC ; about 40 L Isothermal The oven temperature is kept constant during the entire analysis Practical temperature < 250oC Maximum temperature < 400oC Temperature programming The oven temperature is varied during the analysis Linear Non linear High temperature GC (HTGC) Working limit : conventional GC --- 330oC HTGC --- 450oC Masses of analysed substrates: 600-1000

Temperature programming
With homologues, the retention time increases exponentially with the number of carbon. As retention time increases, width increase and the height decreases, making detection impossible after a few peaks have eluted. Since solubility of gas in a liquid decreases as temperatures goes up, we can reduce the retention of a compound by increasing column temperature.

General steps to create a program assuming that the separation is possible 1) Determine initial temperature and time based on best possible separation offirst few peaks 2) Report for the last few peaks to find the best final temperature and time 3) Experiment with various ramps to account for the rest of the components

Temperature programming

Factors to consider :

Variations in solubility of solutes

Changes in volatility of solutes Stability of solutes Flow rate changes Stability of stationary phase

Must stay within Tmin/Tmax of column Other factors are found experimentally

A temperature program
Ex. 40 oC(5 min) 10oC/min 250oC(10 min) C B A

A: initial temperature and holding time B: ramp (oC/min) C: final temperature and holding time
Some GCs will allow for a more complex program.

Raising column temperature

1) 2) Decrease retention time Decrease resolution

3)

Sharpens peaks

Comparison of isothermal and programmed temperature chromatography. Each sample contains linear alkanes run on a 1.6 mm 6 m column containing 3% Apiezon L (liquid phase) on a 100/200 mesh VarAport 30 solid support with He flow rate of 10 ml/min.