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Topic 12 GC and SFC
Topic 12 GC and SFC
Topic 12 GC and SFC
Supercritical fluid chromatography chromatography using a supercritical fluid as the mobile phase and a solid/liquid as a stationary phase
In SFC, the analytes are solvated in the supercritical fluid SFC is applicable to a much wider range of molecules
GC Theory
Mobile-phase flow rates are much higher in GC (pressure drop is much less for a gas) The effect of mobile-phase flow rate on the plate height (H) is dramatic Lower plate heights yield better chromatography However, much longer columns can be used with GC
GC Instrumentation
Basic layout of a GC:
Injector Detector
Carrier Gas
Column Oven
GC Instrumentation
A typical modern GC the Agilent 6890N:
GC Instrumentation
Typical carrier gases (all are chemically inert): helium, nitrogen and hydrogen. The choice of gas affects the detector. Injectors: most desirable to introduce a small plug, volatilize the sample evenly Most samples introduced in solution: microflash injections instantly volatilize the solvent and analytes and sweep them into the column Splitters: effectively dilute the sample, by splitting off a portion of it (up to 1:500) Ovens: Programmable, temperature ranges from 77K (LN2) up to 250 C. Detectors: wide variety, to be discussed shortly
Headspace GC
A very useful method for analyzing volatiles present in non-volatile solids and liquids Sample is equilibrated in a sealed container at elevated temperature The headspace in the container is sampled and introduced into a GC
Needle
Headspace
Liquid/solid
Columns for GC
Open tubular columns: most common, also known as capillary columns (inner diameters of <0.25 mm) up to 150 m long 1000-3000 plates/m pressure limits particle size in packed columns No A term (Eddy or multipath) in van Deemter equation N up to 600000
Packed columns: contain packing, like HPLC columns typical particle sizes 100-600 um 3 m long 1000-3000 plates/m difficult to overload N up to 12000
FSWC: fused-silica wall-coated open tubular columns, very popular in modern applications (a form of WCOT column)
WCOT (GLC): wall-coated open tubular stationary phase coated on the wall of the tube/capillary SCOT (GLC): support-coated open tubular stationary phase coated on a support (such as diatomaceous earth) More capacity that WCOT
The stationary phase is designed to provide a k and that are useful. Polarities cover a wide range (next slide). Stationary phases are usually a uniform liquid coating on the wall (open tubular) or particles (packed) When the polarity of the stationary phase matches that of the analytes, the low-boilers come off first Bonded/cross-linked phases designed for more robust life, less bleeding often these phases are the result of good polymer chemistry
Adsorption onto silicates (via free silanol groups) on the silica column itself: avoided by deactivation reactions, usually leaving an OCH3 group instead.
HO O n
OH
5% phenyl polydimethylsiloxane 50% phenyl polydimethylsiloxane 50% trifluoropropyl polydimethylsiloxane polyethylene glycol
OV-3, SE-52
350
OV-17
250
OV-210
200
Carbowax 20M
250
OV-275
240
High-temperature columns work to 400C, include Agilents DB-1ht (100% polydimethylsiloxane), DB-5ht (5% phenyl).
Temperature Effects in GC
Temperature programming can be used to speed/slow elution, help handle compounds with a wide boiling point range
Comparison of GC Detectors
Detector
Flame ionization (FID) Thermal conductivity (TCD) Electron capture (ECD) Mass spectrometry (MSD) Thermionic (NPD) Electrolytic conductivity (Hall)
Sensitivity
1 pg carbon/sec 500 pg/mL 5 fg/sec 0.25 to 100 pg 0.1 pg/s (P) 1 pg/s (N) 0.5 pg/s (Cl) 2 pg/s (S) 4 pg/s (N)
Selective or Universal
Universal Universal Selective Universal Selective Selective
Common Applications
Hydrocarbons Virtually all compounds Halogens Ionizable species Nitrogen and phosphorus compounds (e.g. pesticides) Nitrogen, sulfur and halogencontaining compounds
Photoionization
Fourier transform IR (FTIR)
2 pg/s
0.2 to 40 ng
Universal
Universal
Compounds ionized by UV
Organics
GC Detectors: FID
The flame ionization detector (FID), the most common and useful GC detector Process: The column effluent is mixed with hydrogen and air and is ignited. Organic compounds are pyrolyzed to make ions and electrons, which conduct electricity through the flame (current is detected) Advantages: sensitive (10-13 g), linear all the way up to 10-4 g), non-selective Disadvantages: Destructive, certain compounds (noncombustible gases) dont give signals in the FID.
GC Detectors: Other
Atomic emission detector: plasma systems (like ICP, but often using microwaves) elemental analysis Sulfur chemiluminescence detector (SCD): reaction between sulfur and ozone, follows an FID-like process Thermionic detector: like an FID, optimized and electrically charged to form a low-temp (600-800 C) plasma on a special bead. Leads to large ion currents for phosphorous and nitrogen a selective detector that is 500x as sensitive as FID Flame photometric detector: specialized form of UV emission from flame products Photoionization detector: UV irradiation used to ionize analytes, detected by an ion current. And, of course, the mass spectrometer (MS)
Interpretation of GC Data
Common use: develop a method to separate compounds of interest by spiking, and use retention times to determine whether a compound is present or not in unknowns
Watch out for compounds with the same retention time! GC can function as a negative test e.g. rule out the presence of ethyl acetate in my sample. Relative retention time:
r (t R ) A /( t R ) std
Quantitative Kovats retention index (I) based on normal alkanes the retention index of these compounds is independent of temperature and packing I = 100z (z is the number of carbons in a compound) Relative retention index:
Results:
ppb detection of VOCs like benzene, decane, halomethanes, etc in water samples
Commercialized by Teledyne Tekmar (e.g. the Velocity XPT) and used worldwide Legally-mandated for water analysis in many areas
Cl
Si CH3
HCl
Si O
H
H O H
testosterone
O
Hyphenation of GC and MS
The first useful hyphenated method? Continuous monitoring of the column effluent by a mass spectrometer or MSD Very easy to interface capillary GC columns have low enough flow rates, and modern MS systems have high enough pumping rates, that GC effluent can be fed directly into the ionization chamber of the MS (for EI or CI, etc) Larger columns require a jet separator Most common systems use quadrupole or ion trap mass analyzers (MSD)
Supercritical Fluids
Phase diagrams show regions where a substance exists in a certain physical state Beyond the critical point, a gas cannot be converted into the liquid state, no matter how much pressure is applied!
Supercritical Fluids
Supercritical properties of CO2
The fluid intermediate between a liquid and a gas Obtained in a not-sosudden manner (there is no real transition)
Supercritical Fluids
Photos of CO2 as it goes from a gas/liquid to a supercritical fluid
1
Meniscus
Increasing temp
Major advantage of SFC over HPLC: SFC can use the universal FID as a detector
SFC can also use UV, IR, and fluorescence detectors SFC is compatible with MS hyphenation
Applications of SFC
Why use SFC over other techniques? Consider speed and capability as well as expense