Analysis of a Liquid Organic Mixture by Gas Chromatography

Introduction

In this experiment, liquid organic samples previously distilled will be analyzed on a

Gas Chromatograph (GC), which allows for the establishment of the identity of the
components in the mixture as well as the quantity of each component.

Objectives:

1. Learn how to operate a Gas Chromatograph.

2. Identify the components of a liquid sample by GC and determine their relative ratio.

Required Reading: Pavia Chapter 22

In order to analyze and quantify the components of liquid samples obtained from the
previous distillation, it will be necessary to find the retention times of each compound in the
mixture and compare them to that of standards (pure compounds). For example, if you
suspect the unknown may contain 1-propanol and toluene, then the retention times for 1-
propanol and toluene will need to be measured. This can be done by injecting a sample of
pure 1-propanol and pure toluene separately into the GC column and then noting the time
required for a peak to appear on the chromatogram (used to describe the graph of a GC run)
relative to the retention time of the air peak. You then analyze your liquid mixture and find
out the retention time for each component (assuming they are separated after going through
the GC column). Since the retention time should be a constant given a set of instrument
condition over a short period of time (lets say within 2-3 hours), the identify of each
component can be assessed by matching the retention time of unknown to the standards. In
a strict sense, this method of identification is only possible if you know the unknown is from
a limited number of probably compounds (such as the list given at the end of the distillation
experiment from which your unknowns were made). If one has no such prior knowledge,
then another instrumental or chemical method may be required in order to positively identify
the unknown. To determine the quantity of each component in the mixture, it will be
necessary to integrate the area under each peak. Several methods of obtaining the area for
each peak are discussed in Pavias book. For our experiment this week, we will use a
GOW-MAC 350 GC connected to electronic integrators that keep track of each peaks
retention time and its area.

Experimental Procedure:

Components of a gas chromatograph There are two columns in our GOW-MAC 350
GC; column A has a polar stationary phase and column B has a non-polar stationary phase.
The oven may be heated to as high as 200 C. The carrier gas used is helium. GOW-MAC
350 is equipped with a thermal conductivity detector (TCD) which gives signal based on the
amount of heat carried away by the sample. All the settings have been fine-tuned by your
instructor for each particular experiment, so there is no need for further adjustment on your
part.

1. Prior to an analysis make sure the instrument is turned on and equilibrated and
that the integrators light indicates that it is ready to plot signal.
2. Make sure the syringe is clean. This can be done by flushing the syringe with a
sample of acetone, moving the plunger up and down several times to force out
any residual acetone. Then flush the syringe 3-4 times with your sample.
3. Next the retention time of the standard will be established. Since your
unknowns were made from the following compounds, each of these compounds
will need to be injected, one at a time, into the GC and the retention time
calculated.

1-propanol, 2-methyl-1-propanol, 1-butanol, toluene, p-xylene,

isopropylbenzene, and mesitylene.

Each group will now inject a pure sample of the above into the GC. Pull in 5-10
ul of air into the syringe, then draw 0.5-1 ul of the pure sample. After the appropriate
amount of sample is in the syringe, pull back the plunger to draw in an additional 5-
10 uL of air. The air peak is carried along rapidly by the carrier gas and acts as a
marker for calculating retention times. Each group will take turn to inject a different
pure sample until the retention time of all 7 pure samples has been obtained.

4. Introduce the sample-air mixture onto the chromatography column by pushing

only 2/3rd of the needle into the injection port (through a rubber septum) and
quickly, but smoothly, injecting the sample into the instrument. During this
operation hold your thumb on the plunger since the positive helium
pressure in the instrument may push the plunger and the sample out of the
syringe.
5. Immediately push the start button on the integrator.
6. While the chromatogram is being traced, wash the syringe with acetone 4-5
times, being sure to dry the syringe by moving the plunger up and down. Place
the syringe back on the bench, ready for the next person to use.
7. Label the chromatogram with the conditions used and the name and amount of
the sample analyzed.
8. On the chromatogram, the integrator will print retention time on each peak (in
the unit of minutes). Wait until all peaks have emerged, then push stop button.
The integrator will print a summary of all peaks monitored. For the standards,
you would only need the corrected retention time of each pure compound
(retention time of compound minus retention time of the air peak which is the
first peak, assuming that enough air has been injected)
9. After the retention times of all standards have been measured, you are ready to
analyze your unknown. Follow the same procedure as above when injecting
your unknown sample (the distillate from the previous experiment).
10. The integrator will print a summary result for your unknown. In this case, you
would want to calculate the corrected retention time (that of each peak minus the
retention time of air) of each component and compare them to the standards. A
match in retention time allows you to determine their identity. To quantify each
component, take the area of that component and divide that by the total area of
all components that are of interest to you. (i.e. excluding the area of air, and
solvent such as acetone or ether) This gives the relative percentage of that
component, and the sum of the relative % of all interested components should
total 100%.
 As always, be sure to record all pertinent data in your laboratory notebook
(retention times of the standards and unknown, and quantitative analysis of each component)
After you establish the identify of the components in your distillate, compare the results to
the probably identity you obtained from the b.p. measurement. Discuss your observations
with your instructor the results before proceeding to dispose of your liquid sample.

Once you are sure that the distillate sample is no longer needed, dispose of them
into the Non-halogenated organic waste and rinse vials 2-3 times with small amount of
acetone. Collect acetone washes in the waste bottle as well. Return the vials to your
instructor.
Lab Report for Analysis of a liquid organic mixture by GC

Name:
Section:

1. Which of the following variables will double in quantity if twice as much sample was
injected into the GC operating under the identical conditions? (Circle one)

(A) The height of the peaks (B) The width of the peaks
(C) The area under each peak (D) None of the above

2. What is the purpose of injecting air into the GC along with the sample ?

3. The retention time of a compound on a GC chromatogram depends upon (circle all that
are true)

(A) Carrier gas flow rate (B) Length of the GC column

(C) Column temperature (D) Boiling point of the compound

4. Complete the following tables:

Standards Retention Time of standards Corrected Retention Time

1-propanol
2-methyl-1-propanol
1-butanol
toluene
p-xylene
isopropylbenzene
mesitylene

First Fraction Retention Time Identity Relative %

Compound 1
Compound 2

Second Fraction Retention Time Identity

Compound 1
Compound 2

5. Are the identity assignments of GC consistent with that of b.p.? Discuss the success of
your distillation in terms of its separation quality based on the relative % above. (use
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