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PRACTICAL # 01

Object: To determine gas composition by gas chromatograph

Theory:
In this experiment, mixtures of volatile organic compounds will be separated and analyzed, and a
sample containing an unknown percentage of ethanol will be quantitated by GC analysis.
In GC, a liquid sample is injected into a separation column as sharp plug. The GC column is usually
coated with stationary phase of a given overall polarity. The column is contained within an oven.
The sample is vaporized to a degree and at a rate dependent on its boiling point. It is carried by an
inert gas (usually helium) through the column to a detector. The detector signals a chart recorder,
which records the response, ideally in the shape of a Gaussian peak.

Fig: 1.1 Gas Chromatograph Schematic Diagram


Two significant factors determine the rate at which an injected compound travels through the
column and reaches the detector. A compound’s volatility at the column temperature influences
the distribution of the compound between the gaseous mobile phase and the stationary phase. Other
things being equal, a compound band will move through the column more quickly if its distribution
favors the mobile phase. Boiling points are often used as a measure of the relative volatilities of
compounds in a mixture. However, volatility alone does not determine the distribution between
the stationary and mobile phase. Specific interactions between the injected compound and the
stationary phase play an important role as well. Recall that “like dissolves like” so a polar
compound will tend to strongly distribute into a polar stationary phase. Conversely, a polar
compound will not have a strong affinity for a non-polar stationary phase and, thus, will elute
relatively quickly. In summary, components in a mixture can be identified by analyzing the
difference in their retention times, which is dependent upon their volatility and polarity.
Quantitation is possible in GC methods by analyzing the peak area or peak height. For the detector
you will use in this experiment, peak area will provide better results. A larger peak area indicates
a larger amount of analytic present in the sample. Spiking samples with internal standards helps to
compensate for the imprecision inherent to GC methods and for variations in sensitivity among
different analyses. In this method, the analytical sample and standards are spiked with an fixed
amount of a solute whose retention time is similar to (but resolved from) that of the sample. The
ratio of the area of the analytic peak to that of the internal standard is used to prepare a calibration
curve and subsequently used to determine the analytic concentration in an unknown mixture.

Fig: 1.2 Gas Chromatograph Setup


Procedure:
In this experiment, you will use a Gown-Mac GC equipped with two columns. Column A is a polar
column with a tradename of Carbora 20M and column B is a non-polar column with a tradename
of DC 200. Only one column may be used at a time. The direction that the chart recorder pen will
deflect depends upon which column is in use; make certain that you change the polarity of the pen
direction whenever you change columns.
Several factors affect separation efficiency in GC, including column temperature and carrier gas
flow rate. The temperature for the GC oven has been preset for you; record the temperature in your
lab notebook. The carrier gas flow rate is adjustable for both columns and, thus, the flow rate will
not be the same for both columns. You will use a soap bubble flowmeter to measure the flow rate
of the carrier gas. 1 Before starting, the detector and chart recorder must be zeroed. Set the
Attenuation to infinity and position the chart recorder pen to the desired baseline position using
the recorder Zero offset. Slowly turn the Attenuation control towards 1. If the chart recorder pen’s
position moves from the baseline setting, use the Zero control on the GC to reset the pen to its
original position. Once you are able to change the attenuation from infinity to 1 without
significantly changing the pen’s position, the instrument is zeroed. Begin your experiment by
setting the attenuation on 4 and the chart recorder sensitivity to 20 mV. Adjusting these parameters
will change the peak size on your chart paper. Setting the recorder’s sensitivity to a higher setting
will cause the peaks to become smaller; increasing the attenuation to a larger number will cause
the peaks to become dramatically smaller. Set the chart recorder speed to 5 cm/min. The TA will
demonstrate the proper use of the GC injection micro syringes. Treat these expensive devices with
care! Whenever a sample is injected into the GC, it is important to mark the chart paper at the
moment of injection. Also, you should inject as much air as possible with the sample in order to
obtain an air peak (this allows determination of adjusted retention times). The following chart lists
some important properties of the classes of compounds used in this experiment: Compound
Polarity Boiling Point alcohols polar boiling point increases with molecular weight ketones polar
lower boiling point than alcohols of comparable molecular weight alkanes non-polar boiling point
is lower than ketones of comparable molecular weight alkyl benzenes non-polar high degree of
double bonds increases stability, thus, relatively high boiling points Procedure: Sample Prep 1.
Each group needs one of each of the following solutions: Mix A: equal amount of straight chain
alcohols from C2-C6 Mix B: equal amounts of n-heptane, 2-butanone, 1-butanol and o-xylene pure
n-heptane pure 2-butanone pure 1-butanol pure o-xylene Each student must obtain an individual
unknown. 2 2. Prepare the following standards in a 10-mL volumetric flask. Fill the flasks to the
mark with water after adding the ethanol and n-butanol. (volumes in mL): % ethanol (v/v) vol.
ethanol vol. n-butanol 20% 2.00 1.00 40% 4.00 1.00 60% 6.00 1.00 80% 8.00 1.00 3. Pipette 5 mL
of your unknown into a 10-mL volumetric flask. Add 1 mL n-butanol and fill to the mark with
water. Procedure: Chromatography For each set of chromatograms, record the following: solution
injected recorder sensitivity (mV) flow rate (mL/min) chart speed injection volume (µL) column
temperature (o C) type of column GC attenuation 4. Inject 1 µL of the alcohol mixture onto column
A. Inject as much air as possible with the sample. Adjust parameters as necessary until retention
times for each peak can be analyzed. 5. Individually analyze each compound in mix B on each
column by injecting 2 µL of each pure substance. REMEMBER: you cannot operate both columns
simultaneously. 6. Inject 1 µL of mix B onto each column. 7. Inject 1 µL of the ethanol standards
onto column A. Adjust parameters so that the peak areas can be easily measured. 8. Inject 1 µL of
the unknown onto column A; repeat 3-4 times.

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