Chem 223: Experiment 13

GC/FID Determination of Toluene in Air

References:
1. Harris. Chap. 24
2. Kealey. Pp. 137-154
3. Harvey. 12D
4. http://teaching.shu.ac.uk/hwb/chemistry/tutorials/chrom/gaschrm.htm.
This experimental procedure is based on the experiment on website:
http://web.pdx.edu/~atkinsdb/teach/427/Expt-GCTolAir.pdf

I. Purpose of the experiment

In this experiment you will measure the volume fraction concentration (in parts per billion by
volume, ppbv) of toluene in air samples that you will collect. The samples will be collected and
stored in Teflon bags which will be returned to the laboratory for analysis. The method of
analysis is gas chromatography with cryogenic preconcentration and flame ionization detection
(GC/FID). The quantitation method will be based on an internal standard using m-xylene and a
standard addition of toluene. Several air samples will be analyzed.

Your group will first go out into the “real world” and draw a few (three to four) air samples, and
bring them back to the laboratory for analysis. You should try to collect samples that will show a
variation in hydrocarbon pollution content. Take notes on where you took the samples and the
relevant atmospheric conditions.

You’ll run two gas chromatograms per sample (raw and spiked, both with the internal standard)
except for the first sample, where you will also run an experiment without cryogenic
preconcentration. The non-preconcentrated run can be compared to the same sample with
preconcentration to determine the extent of the preconcentration. The time per analysis (each
chromatographic analysis will be referred to as a “run”) is over 20 minutes, so you need to be on
the ball to get three or more air samples done during the lab period.

You will be pumping the air samples through a small (~ 100 μL) metal sample loop that is
immersed in liquid nitrogen (the cryogen) for a pre-planned length of time, usually about 5
minutes. The N2 doesn’t affect the majority components of air (nitrogen and oxygen) under our
conditions, so they flow on through to the air pump. The hydrocarbons (and water and carbon
dioxide) are trapped in the sampling loop during the passage. Theoretically, the longer you run the
air through the cryogenically cooled loop, the more hydrocarbon you will end up with in the loop;
but in the interest of time, we won’t test that idea.

After the preconcentration (or shortly after the flow is established in the non-preconcentrated
sample) a sample valve similar to those used in liquid chromatography is used to inject the
volatile contents of the sample loop onto the column for separation and eventual detection and
measurement by a flame ionization detector (FID).

II.Introduction:
 Gas chromatography - specifically gas-liquid chromatography - involves a sample being
vapourised and injected onto the head of the chromatographic column. The sample is
transported through the column by the flow of inert, gaseous mobile phase. The column itself
contains a liquid stationary phase which is adsorbed onto the surface of an inert solid.
The picture and schematic diagram of a gas chromatography are following.

Fig. GC ( Shimadzu) basic system

Instrumental components
Carrier gas
The carrier gas must be chemically inert. Commonly used gases include nitrogen, helium, argon,
and carbon dioxide. The choice of carrier gas is often dependant upon the type of detector
which is used. The carrier gas system also contains a molecular sieve to remove water and other
impurities.

Sample injection port

For optimum column efficiency, the sample should not be too large, and should be introduced
onto the column as a "plug" of vapor - slow injection of large samples causes band broadening
and loss of resolution. The most common injection method is where a microsyringe is used to
inject sample through a rubber septum into a flash vapouriser port at the head of the column.
The temperature of the sample port is usually about 50C higher than the boiling point of the
least volatile component of the sample. For packed columns, sample size ranges from tenths of
a microliter up to 20 microliters. Capillary columns, on the other hand, need much less sample,
typically around 10-3 L. For capillary GC, split/splitless injection is used. Have a look at this
diagram of a split/splitless injector;
The injector can be used in one of two modes; split or splitless. The injector contains a heated
chamber containing a glass liner into which the sample is injected through the septum. The
carrier gas enters the chamber and can leave by three routes (when the injector is in split mode).
The sample vaporizes to form a mixture of carrier gas, vaporized solvent and vaporized solutes.
A proportion of this mixture passes onto the column, but most exits through the split outlet. The
septum purge outlet prevents septum bleed components from entering the column.

Columns
There are two general types of column, packed and capillary (also known as open tubular).
Packed columns contain a finely divided, inert, solid support material (commonly based on
diatomaceous earth) coated with liquid stationary phase. Most packed columns are 1.5 - 10m in
length and have an internal diameter of 2 - 4mm.
Capillary columns have an internal diameter of a few tenths of a millimeter. They can be one of
two types; wall-coated open tubular (WCOT) or support-coated open tubular (SCOT).
Wall-coated columns consist of a capillary tube whose walls are coated with liquid stationary
phase. In support-coated columns, the inner wall of the capillary is lined with a thin layer of
support material such as diatomaceous earth, onto which the stationary phase has been adsorbed.
SCOT columns are generally less efficient than WCOT columns. Both types of capillary
column are more efficient than packed columns.
In 1979, a new type of WCOT column was devised - the Fused Silica Open Tubular (FSOT)
column;
These have much thinner walls than the glass capillary columns, and are given strength by the
polyimide coating. These columns are flexible and can be wound into coils. They have the
advantages of physical strength, flexibility and low reactivity.

Column temperature
For precise work, column temperature must be controlled to within tenths of a degree. The
optimum column temperature is dependant upon the boiling point of the sample. As a rule of
thumb, a temperature slightly above the average boiling point of the sample results in an elution
time of 2 - 30 minutes. Minimal temperatures give good resolution, but increase elution times.
If a sample has a wide boiling range, then temperature programming can be useful. The column
temperature is increased (either continuously or in steps) as separation proceeds.
Detectors
There are many detectors which can be used in gas chromatography. Different detectors will
give different types of selectivity. A non-selective detector responds to all compounds except
the carrier gas, a selective detector responds to a range of compounds with a common physical
or chemical property and a specific detector responds to a single chemical compound. Detectors
can also be grouped into concentration dependant detectors and mass flow dependant detectors.
The signal from a concentration dependant detector is related to the concentration of solute in
the detector, and does not usually destroy the sample Dilution of with make-up gas will lower
the detectors response. Mass flow dependant detectors usually destroy the sample, and the
signal is related to the rate at which solute molecules enter the detector. The response of a mass
flow dependant detector is unaffected by make-up gas. Have a look at this tabular summary of
common GC detectors:
Support Dynamic
Detector Type Selectivity Detectability
gases range
Flame ionization Hydrogen
Mass flow Most organic cpds. 100 pg 107
(FID) and air
Thermal conductivity
Concentration Reference Universal 1 ng 107
(TCD)
Halides, nitrates,
Electron capture nitriles, peroxides,
Concentration Make-up 50 fg 105
(ECD) anhydrides,
organometallics
Hydrogen Nitrogen,
Nitrogen-phosphorus Mass flow 10 pg 106
and air phosphorus
Sulphur,
Hydrogen phosphorus, tin,
Flame photometric and air boron, arsenic,
Mass flow 100 pg 103
(FPD) possibly germanium,
oxygen selenium,
chromium
Aliphatics,
aromatics, ketones,
Photo-ionization
Concentration Make-up esters, aldehydes, 2 pg 107
(PID)
amines,
heterocyclics,
 organosulphurs,
some
organometallics
Halide, nitrogen,
Hall electrolytic Hydrogen,
Mass flow nitrosamine,
conductivity oxygen
sulphur

The effluent from the column is mixed with hydrogen and air, and ignited. Organic compounds
burning in the flame produce ions and electrons which can conduct electricity through the flame.
A large electrical potential is applied at the burner tip, and a collector electrode is located above
the flame. The current resulting from the pyrolysis of any organic compounds is measured.
FIDs are mass sensitive rather than concentration sensitive; this gives the advantage that
changes in mobile phase flow rate do not affect the detector's response. The FID is a useful
general detector for the analysis of organic compounds; it has high sensitivity, a large linear
response range, and low noise. It is also robust and easy to use, but unfortunately, it destroys the
sample.

III. Reagent and Apparatus:

- Shimadzu Gas Chromatograph with FID detection and air sample loop injection Tedlar (a type
of Teflon) gas sampling bags with valve and septum port “Bucket brigade” air sampling apparatus
Dewar and liquid nitrogen 1.0mL Pressure-Loc Gas Syringe.

Standards Available:

~100ppmv m-xylene air standard bag

~100ppmv toluene air standard bag

(Ask your TA for the data needed to calculate the exact concentration of the standards.)

IV. Experimental Procedure:

A. Air sampling
• Have the TA show you how to use the sampling bucket and Tedlar bags.

• Go out into the “field” and take a few air samples - i.e., fill up three or four Tedlar bags with
air. Air samples taken in this way are referred to as “grab” samples. Try and fill the bags as
completely as possible.

• To make things interesting, try taking samples from areas where you expect there to be a lot of
hydrocarbons (high traffic) and only a little bit of pollution (nearly the city-wide background.)
Keep records of where you took each sample from and other observations (e.g., heavy traffic,
wind from South, temperature, barometric pressure, raining…)

• Bring all samples back to the laboratory for analysis.

B. Internal Standard/Standard Addition Procedure

For each air sample, we will run one chromatogram of the raw sample with m-xylene and one of a
toluene spiked sample with the internal standard. (We’re making the assumption here that
m-xylene isn’t present in ambient air.). To add m-xylene or toluene to the air samples:

• Insert the 1.0 mL graduated gas syringe into the standard bag through the septum in the Tedlar
bag (the tiny hole on the side of the valve/inlet for the bag).

• Press the green button to open the syringe valve.

• Rinse the syringe a few times and then pull a 1.0mL sample of the gas standard.

• Press the red button to close the valve.

• Pull the syringe out of the standard bag and insert it through the septum port in the sample gas
bag.

• Press the green button.

• Inject the spike and remove the syringe.

• Mix the sample thoroughly by “squishing” the bag for a few minutes at least. {It’s boring and
noisy, but if you don’t work at it, you will not end up with good results because the diffusional
mixing of gases at atmospheric pressure is very slow.}

• Measure the volume of the bag and calculate the concentration of m-xylene or toluene. (We are
currently measuring volume using a water displacement method. See the Appendix for
directions)

• Put the internal standard in all of the sample bags before you start the analyses.

C. Analysis:

• Make sure that the computer and the GC are on and that the FID is lit and has stabilized.

• Get a transfer dewar full of liquid nitrogen from the big stainless steel storage dewar. (If you
haven’t done this before, ask the TA for help.)

For the non-preconcentrated run

• Choose the Tedlar bag that you think will have the highest toluene concentration and insert the
 valve stem into the flexible tubing coming out of the center of the sampling valve on the front
of the GC. (It’s a tight fit, but you don’t have to have more than a few mm on to make a seal.)

• Make sure the sampling valve is in the LOAD position, turn on the air pump, open the valve on
the bag (push it in), and allow the sample to flow through the loop for a few seconds.

• Switch the sampling valve to the INJECT position and press the START button on the GC.
Within a few seconds, a red vertical line should appear on the continuous output screen on the
computer, signifying the beginning of a chromatogram.

• Close the valve (pull it out) on the Tedlar bag and remove it from the flexible tubing.

• After ~ 1 minute, return the sampling valve to the LOAD position to get ready for the next run.
{You can leave the air pump on all the time, to help flush the last sample from the sampling loop
before the next chromatogram.}

• The run takes 18.75 minutes and the toluene and m-xylene elute well before the end, but you
should always let the chromatograph run for the full time to drive higher molecular weight
compounds from the column.

• When the chromatogram finishes, the status bar on the computer will change from BLUE
(running) to RED (not ready) and the temperature of the oven will start to go down. The
chromatogram will briefly appear on the computer screen (then the screen will go back the
running signal display) and the Report Screen will refresh. {At this point, you can print the
report by selecting Tabulate Print on the top of the Report Screen.}

For ALL preconcentrated runs (i.e., all the rest of the analyses)

• Attach the Tedlar bag’s valve to the flexible tubing on the pump.

• Make sure the sampling valve is in the LOAD position and turn on the air pump, open the valve
on the bag (push it in), and allow the sample to flow through the loop for a few seconds.

• Next you will simultaneously place a Styrofoam cup full of liquid nitrogen around the sample
loop, immersing the bottom half of the coils in the cryogen, and start a timer. {The length of
time that the cryogen is on the loop determines the extent of preconcentration, and thus should
be as consistent between a sample and its spiked version as possible. You could change the
trapping time between different samples if more preconcentration was desired, but this is
usually not necessary and can cause confusion.}

Read through the next three steps as they are all time critical

• At the end of the predetermined trapping time, usually around 5 minutes, remove the cup with
the liquid nitrogen cryogen and wait for ~3 seconds to allow liquid oxygen to escape from the
sampling loop

• then switch the sampling valve to the INJECT position

• then place a cup full of room temperature (or warmer) water around the loop and press the
START button on the GC. {The water heats the loop and rapidly drives the volatile contents
out of the loop and onto the GC column.}

• Close the valve on the gas sampling bag, remove it from the flexible tubing, and measure the
new air bag volume.
 After ~ 1 minute, return the sampling valve to the LOAD position to get ready for the next run.

Again, when the chromatogram finishes, it will briefly appear on the computer screen (then the
screen will go back the running signal display) and the Report Screen will refresh. {At this
point, the chromatogram and peak integrations should print automatically. If not, you can print
this information by selecting Tabulate Print on the top of the Report Screen. Don’t forget to
print before the next chromatogram finishes, or you will lose your data.}

After the chromatogram of the raw sample (with m-xylene) is done, and you’ve measured the
volume of the air in the bag, and you have the report printout in hand, spike the sample with
toluene. {This last part seems silly, since it would be better to measure the volume of the bag and
spike it immediately after the “raw” injection so that the spike would have more time to mix into
the air in the bag. But you can’t “unspike” a bag, and if anything goes wrong during a run and
you don’t get the data from the “raw” sample, there is no point in running the spiked version. Up
to you, but this is the safest way.}

Analyze the spiked sample as you did with the raw one {start over again at For ALL
preconcentrated runs…} and obtain the report printout. Locate the toluene and m-xylene peaks
in the “raw” and spiked runs. (The relative peak area of toluene should have increased.) {Also,
retention times should be nearly the same, but the toluene peak isn’t the only one that changes}
and obtain the peak areas. Calculate the toluene concentration of the air sample using the
volume of the standard and sample bags and the ideal gas law.

FOR YOUR REPORT:

In writing this report, you should include the following:

1. Present the relevant peak areas, concentrations, and quantitative results (in ppbv toluene) in
tabular form.

2. Be sure to explain in the Introduction how toluene concentration was calculated.

3. Comment on the extent of preconcentration, based on your sample run without the cryogen -
quantitatively if possible (i.e., you have to have seen something to figure out how much it
increases - otherwise the answer might just be “a lot”.)

4. Provide a map of your sampling locations and say whether your results were consistent with
your expectations during sampling.

5. In addition to quoting the volume fraction concentration for the “unknown” air samples, also
give the ppmvC (volume fraction carbon.) This is the relevant parameter in calculating the
ozone producing potential of VOCs.

ITEMS TO THINK ABOUT:

1. The GC you used was equipped with a thermal conductivity detector. What are the strengths
and weaknesses of this kind of detector?

2. Why is temperature programming sometimes useful in gas chromatography?

3. What changes would you expect in retention time, peak shape, and resolution if a. You
decreased the column temperature between replicate injections of the same mixture?
3. You increased the mobile phase flow rate between replicate injections of the same mixture?