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Mobile phases are generally inert gases such as helium, argon, or nitrogen. The inject ion
Port consists of a rubber septum through which a syringe needle is inserted to inject the
sample. The injection port is maintained at a higher temperature than the boiling point of
the least volatile component in the sample mixture. Since the partitioning behavior is
dependant on temperature , the separation column is usually contained in a thermostat-
controlled oven. Separating components with a wide range of boiling points is
accomplished by starting at a low oven temperature and increasing the temperature over
time to elute the high-boiling point components. Most columns contain a liquid stationary
phase on a solid support. Separation of low-molecular weight gases is accomplished with
solid adsorbents. Separate documents describe some specific GC Columns and GC
Detectors.
Schematic of a gas chromatograph
(purified silicate glass) capillary (typically 10-100 m in length and 250 µm inner
diameter) that has the stationary phase coated on the inner surface. Capillary columns
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provide much higher separation efficiency than packed columns but are more easily
overloaded by too much sample.
Specific GC detectors
Chemiluminescence detector
As capillary column based gas chromatography takes its place as the major, highest
resolution separation technique available for volatile, thermally stable compounds, the
requirements for the sensitive and selective detection of these compounds increases.
Since more and more complex mixtures can be successfully separated, subsequent
chromatograms (output of a chromatographic separation) are increasingly more complex.
Therefore, the need to differentiate between the sample components using the GC
detector as a means of compounds discriminating is more and more common. In addition,
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each detector has its own characteristics (selectivity, sensitivity, linear range, stability,
cost, etc.) that helps in a decision about which detector to use.
ne of the newest additions to the gas chromatographer's arsenal is the atomic emission
detector (AED). This detector, while quite expensive compared to other commercially
available GC detectors, is an extremely powerful alternative. F INSTANCE, Instead
of measuring simple gas phase (carbon containing) ions created in a flame as with the
flame ionization detector, or the change in background current because of electronegative
element capture of thermal electrons as with the electron capture detector, the AED has a
much wider applicability because it is based on the detection of atomic emissions.
The strength of the AED lies in the detector's ability to simultaneously determine the
atomic emissions of many of the elements in analytes that elute from a GC capillary
column (called eluants or solutes in some books). As eluants come off the capillary
column they are fed into a microwave powered plasma (or discharge) cavity where the
compounds are destroyed and their atoms are excited by the energy of the plasma. The
light that is emitted by the excited particles is separated into individual lines via a
photodiode array. The associated computer then sorts out the individual emission lines
and can produce chromatograms made up of peaks from eluants that contain only a
specific element.
Instrumentation
The components of the AED include 1) an interface for the incoming capillary GC
column to the microwave induced plasma chamber, 2) the microwave chamber itself, 3) a
cooling system for that chamber, 4) a diffraction grating and associated optics to focus
then disperse the spectral atomic lines, and 5) a position adjustable photodiode array
interfaced to a computer. The microwave cavity cooling is required because much of the
energy focused into the cavity is converted to heat.
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Chemiluminescence Spectroscopy
Introduction
Chemiluminescence, like atomic emission spectroscopy (AES), uses quantitative
measurements of the opticalemis s ion from excited chemical species to determine analyte
concentration; however, unlike AES, chemiluminescence is usually emission from
energized molecules instead of simply excited atoms. The bands of light determined by
this technique emanate from molecular emissions and are therefore broader and more
complex then bands originating from atomic spectra. Furthermore, chemiluminescence
can take place in either the solution or gas phase, whereas AES is almost strictly as gas
phase phenomenon.
Though liquid phase chemiluminescence plays a significant role in laboratories using this
analytical technique (often in conjunction with liquid chromatography
), we will
concentrate on gas phase chemiluminescence reactions since the instrumental
components are somewhat simpler. These detectors are also often used as detectors for
gas chromatography.
Like fluorescence spectroscopy, chemiluminescence's strength lies in the detection of
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electromagnetic radiation produced in a system with very low background. And on top of
this, because the energy necessary to excite the analytes to higher electronic, vibrational,
and rotational states (from which they can decay be emission) does not come from an
external light source like a laser or lamp, the problem of excitation source scattering is
completely avoided. The major limitation to the detection limits achievable by
chemiluminescence involves the dark current of the photomultip lier (PMT) necessary to
detect the analyte light emissions.
If the excitation energy for analytes in chemiluminescence doesn't come from a source lamp or laser, where does it come from?
The energy is produced by a chemical reaction of the analyte and a reagent. An example of a reaction of this sort is shown
below:
A chemiluminescence reaction
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To the analytical chemist the ambiguity about the actual products in the reaction is, in
most case, not important. All the analyst cares about is the sensitivity of the instrument
(read detection limits for target analytes), its selectivity-that is, response for an analyte as
compared to an interfering compound, and the linear range of response.
uere is a schematic of the components necessary for a gas phase chemiluminescence
detector interfaced to a capillary gas chromatograph.
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two kinds of atoms. The emitting species for sulfur compounds is excited S2. The lambda
max for emission of excited S2 is approximately 394 nm. The emitter for phosphorus
compounds in the flame is excited HP (lambda max = doublet 510-526 nm). In order to
selectively detect one or the other family of compounds as it elutes from the GC column
an interference filter is used between the flame and thephotomultiplier tube (PMT) to isolate the appropriate emission band. The
drawback here being that the filter must be exchanged between chromatographic runs if the other family of compounds is to be
detected.
Instrumentation
In addition to the instrumental requirements for 1) a combustion chamber to house the
flame, 2) gas lines for hydrogen (fuel) and air (oxidant), and 3) an exhaust chimney to
remove combustion products, the final component necessary for this instrument is a
thermal (bandpass) filter to isolate only the visible and UV radiation emitted by the
flame. Without this the large amounts of infrared radiation emitted by the flame's
combustion reaction would heat up the PMT and increase its background signal. The
PMT is also physically insulated from the combustion chamber by using poorly
(thermally) conducting metals to attach the PMT housing, filters, etc.
The physical arrangement of these components is as follows: flame (combustion)
chamber with exhaust, permenant thermal filter (two I filters in some commercial
designs), a removable phosphorus or sulfur selective filter, and finally the PMT.
Schematic of a gas chromatographic flame photometric detector
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Photoionization Detector
Introduction
The reason to use more than one kind of detector for gas chromatography is to achieve
selective and/or highly sensitive detection of specific compounds encountered in
particular chromatographic analyses. The selective determination of aromatic
hydrocarbons or organo-heteroatom species is the job of the photoionization detector
(PID). This device uses ultraviolet light as a means ofionizing an analyte exiting from a
GC column. The ions produced by this process are collected by electrodes. The current
generated is therefore a measure of the analyte concentration.
Theory
If the energy of an incoming photon is high enough (and the molecule is quantum
mechanically "allowed" to absorb the photon) photo-excitation can occur to such an
extent that an electron is completely removed from its molecular orbital, i.e. ionization.
A Photoionization Reaction
If the amount of ionization is reproducible for a given compound, pressure, and light
source then the current collected at the PID's reaction cell electrodes is reproducibly
proportional to the amount of that compound entering the cell. The reason why the
compounds that are routinely analyzed are either aromatic hydrocarbons or heteroatom
containing compounds (like organosulfur or organophosphorus species) is because these
species have ionization potentials (IP) that are within reach of commercially available
UV lamps. The available lamp energies range from 8.3 to 11.7 ev, that is, lambda max
ranging from 150 nm to 106 nm. Although most PIDs have only one lamp, lamps in the
PID are exchanged depending on the compound selectivity required in the analysis.
Selective detection using a PID
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Here is an example of selective PID detection: Benzene's boiling point is 80.1 degrees C
and its IP is 9.24 ev. (Check the C C Handbook 56th ed. page E-74 for IPs of common
molecules.) This compound would respond in a PID with a UV lamp of 9.5 ev
(commercially available) because this energy is higher than benzene's IP (9.24).
Isopropyl alcohol has a similar boiling point (82.5 degrees C) and these two compounds
might elute relatively close together in normal temperature programmed gas
chromatography, especially if a fast temperature ramp were used. However, since
isopropyl alcohol's IP is 10.15 ev this compound would be invisible or show very poor
response in that PID, and therefore the detector would respond to one compound but not
the other.
Instrumentation
Since only a small (but basically unknown) fraction of the analyte molecules are actually
ionized in the PID chamber, this is considered to be a nondestructive GC detector.
Therefore, the exhaust port of the PIDcan be connected to another detector in series with
the PID. In this way data from two different detectors can be taken simultaneously, and
selective detection of PID responsive compounds augmented by response from, say, an
FIDor ECD. The major challenge here is to make the design of the ionization chamber
and the downstream connections to the second detector as low volume as possible (read small diameter) so that peaks that
have been separated by the GC column do not broaden out before detection.
Schematic of a gas chromatographic photoionization detector
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Two pairs of TCDs are used in gas chromatographs. ne pair is placed in the column
effluent to detect the separated components as they leave the column, and another pair is
placed before the injector or in a separate reference column. The resistances of the two
sets of pairs are then arranged in a bridge circuit.
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The bridge circuit allows amplification of resistance changes due to analytes passing over the sample thermoconductors and
does not amplify changes in resistance that both sets of detectors produce due to flow rate fluctuations, etc.
diluted before it was analyzed. therwise the column would beoverload ed by those high
concentrated components. An example of this appears in the first figure below. The
clearly overload peaks are indicated. And while some of the other components are in the
resolvable (not overloaded) range, having large masses of components can also distort the
peak shape of some of the lower mass components.
An Overloaded Chromatogram
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The following figure shows a little better chromatography with fewer overloaded peaks.
The second eluting peak (about 6 minutes) is clearly not overloaded while the group
between 10 and 14 minutes still shows overloading characteristics: long drawn-out tailing
and much less than baseline separation with peaks that elute nearby (the 11 and 12
minute peaks, for instance).
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ne last subtle point about the configuration of the carrier gas inlet: notice that it enters
the injector at about the middle of the heating block and its gas has to travel along the
outside of the injector before it enters the injection port liner AT THE TP. This is so the
carrier is preheated before it enter the liner where the sample is vaporized. This helps to
prevent a cold spot at the top of the injector where the carrier gas enters.
Now remember that the size of the capillary column limits the amount of analyte that can
be injected, otherwise, chromatographic overloading occurs. Therefore this packed
column injector design, if used with a capillary column, would require that samples with
high concentrations of analytes be diluted. Unless... what other alternative is there to get
the amount of analytes that are injected onto the column smaller without having to dilute
concentrated samples? The solution is the split/splitless capillary GC injector.
The Split/Splitless Capillary Injector
so far so good. But how does the split/splitless injector work? It starts with the same
requirements as the packed column injectors: carrier gas inlet, a septum, septum purge,
injector insert, heater block, and column connection; but the heart of this technological
feat is another set of gas linesout of the injector-another path that the vaporized sample
can take. This is called the split line or vent. The manufacturers of these systems design
them so that the carrier gas flow onto the column is constant-to maintain the
chromatographic requirements of the column and yield reproducible retention times for
analytes. At the same time, the amount of gas that goes out the split vent controls the
amount of sample that enters the column. If the split vent is closed, via a computer
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controlled split valve, then all of the sample introduced into the injector goes on the
column. If the split vent is open then most of the vaporized sample is thrown away to
waste via the split vent and only a small portion of the sample is introduced to the
column. The following diagram illustrates a split/splitless injector with the split venton
so that only a small portion of the sample injected goes on the column
And finally, a very neat aspect of this is that the amount of gas exiting the split vent can
be varied while keeping the flow onto the column constant. This means that the
AMUNT of the split (called the split ratio) can be varied. A common split ratio is 50 to
1. That is, for every 50 units of gaseous sample that are thrown away to waste, 1 unit
goes on the column.
The analyst keeps careful control of the split ratio so that results from the
chromatography can still be quantified. Chromatographic peaks that show up as, say 2.5
ng of compound X really represent 2.5 x 50 = 125 ng of analyte X in the original sample.
Also notice that this mass (125 nanograms) would have overloaded the column if all of it
ended up on the capillary column. Voila! A split injection, and no sample dilution
required.
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