Gas Chromatography: Analysis of Recreational Alcohol

1. Define gas chromatography. A form of chromatography in which the mobile

phase is a gas. Gaseous analyte is transported through the column by a
gaseous mobile phase, called the carrier gas.
2. What are some advantages for using a GC? Quantitative analysis using GC has
advantages including its excellent sensitivity and wide range of applicability.
Chromatographic conditions can be easily manipulated to optimize the
separation and offer potential freedom from interferences.
3. Describe the components of the GC instrument. Volatile liquid or gaseous
sample is injected through a septum, a rubber disk, into a headed port, in
which it rapidly evaporates. Vapor is swept through the column by He, N2, or
H2 carrier gas, and separated analytes flow through a detector, whose
response is displayed on a computer. The column must be hot enough to
provide sufficient vapor pressure for analytes to be eluted in a reasonable
time. The detector is maintained at a higher temperature than the column so
analytes will be gaseous.
4. How are components separated while passing through the column? The
temperature is raised so that the sample rapidly evaporates and separates.
5. What is your internal standard? 1-butanol
6. Explain the internal standard method.

An internal standard in analytical chemistry is a chemical substance that is added in

a constant amount to samples, the blank and calibration standards in a chemical
analysis. This substance can then be used for calibration by plotting the ratio of the
analyte signal to the internal standard signal as a function of the analyte
concentration of the standards. This is done to correct for the loss of analyte during
sample preparation or sample inlet. The internal standard is a compound that is
very similar, but not identical to the chemical species of interest in the samples, as
the effects of sample preparation should, relative to the amount of each species, be
the same for the signal from the internal standard as for the signal(s) from the
species of interest in the ideal case. Adding known quantities of analyte(s) of
interest is a distinct technique called standard addition, which is performed to
correct for matrix effects.

This ratio for the samples is then used to obtain their analyte concentrations from a
calibration curve. The internal standard used needs to provide a signal that is
similar to the analyte signal in most ways but sufficiently different so that the two
signals are readily distinguishable by the instrument. For example deuterated
chlorobenzene (C6D5Cl) is an internal standard used in the analysis of volatiles on
GC-MS because it is similar to Chlorobenzene but does not occur naturally.
Norleucine is also a popular internal standard for the analysis of amino acids via GC-
MS.

7. How does your internal standard make your analysis more accurate?
Theoretically, when the composition of the analyte is totally unknown, the sample is
injected to get the peaks and assuming that the detector response ration of all the
components is equal, the sum of areas of all the peakes is taken as 100%. Based on
this every peak is aestimated for its purity. When tghe composition is known, the
purity is calculated by comparison with that of known standard. This is aclled
external standard method. In order to take care of manual error of injection volume,
the internal stanadrd method is used so that even though there is an error in the
injection volume, the ratio of peak area of sample and that of internal standard does
not change and the answer is more accurate and reliable.

8. Describe the detector used in this experiment. A flame ionization detector

(FID):

In order to detect these ions, two electrodes are used to provide a potential
difference. The positive electrode doubles as the nozzle head where the flame is
produced. The other, negative electrode is positioned above the flame. When
first designed, the negative electrode was either tear-drop shaped or angular
piece of platinum. Today, the design has been modified into a tubular electrode,
commonly referred to as a collector plate. The ions thus are attracted to the
collector plate and upon hitting the plate, induce a current. This current is
measured with a high-impedance picoammeter and fed into an integrator. The
manner in which the final data is displayed is based on the computer and
software. In general, a graph is displayed that has time on the x-axis and total ion
on the y-axis.

The current measured corresponds roughly to the proportion of reduced carbon

atoms in the flame. Specifically how the ions are produced is not necessarily
understood, but the response of the detector is determined by the number of
carbon atoms (ions) hitting the detector per unit time. This makes the detector
sensitive to the mass rather than the concentration, which is useful because the
response of the detector is not greatly affected by changes in the carrier gas flow
rate.
The eluent exits the GC column and enters the FID detector’s oven. The oven is
needed to make sure that as soon as the eluent exits the column, it does not
come out of the gaseous phase and deposit on the interface between the column
and FID. This deposition would result in loss of eluent and errors in detection. As
the eluent travels up the FID, it is first mixed with the hydrogen fuel and then
with the oxidant. The eluent/fuel/oxidant mixture continues to travel up to the
nozzle head where a positive bias voltage exists. This positive bias helps to repel
the reduced carbon ions created by the flame pyrolyzing the eluent. The ions are
repelled up toward the collector plates, which are connected to a very sensitive
ammeter, which detects the ions hitting the plates, then feeds that signal to an
amplifier, integrator, and display system. The products of the flame are finally
vented out of the detector through the exhaust port.

9. Which gas is used as the carrier gas? Helium

10. What are some of the parameters that can be adjusted to achieve better
separation? Flow rate, temperature
11. What property of the compounds in this experiment determines their elution
order? The size of the compound (mass). The smaller the compound, the
faster the elution.
12. Explain what a response factor is and how it is used in to calculate the
concentrations of the compounds in this experiment. Response factor,
usually in chromatography and spectroscopy, is the ratio between a signal
produced by an analyte, and the quantity of analyte, which produces the
signal. Ideally, and for easy computation, this ratio is unity (one). In real-
world scenarios, this is often not the case.
13. Why are we increasing the temperature during a sample run? To allow for
the higher mass/larger compounds to evaporate and be detected.