CHNG 1001 - APPLIED CHEMISTRY 1

GAS CHROMATOGRAPHY

OBJECTIVES

1) To determine the percentage ethanol in various brands of beer.

2) To compare the calculated percentages with the value stated on the respective labels.

THEORY
Principles
Gas Chromatography (GC) is a separation technique where volatile, thermally stable solutes are
separated by sample distribution between two phases; a stationary phase and a mobile phase. The
stationary phase can either be high boiling liquids on an inert support, waxes or solid adsorbents.
The mobile phase is a chemically inert gas (usually nitrogen or helium) that transports the
analyte molecules through the heated column. Solutes are generally eluted in order of increasing
boiling points or decreasing polarity. The column is enclosed in a thermostatically-controlled
oven that is maintained at a steady temperature or programmed to increase progressively during
separation. The detector generates an electrical signal that can be amplified and presented in the
form of a chromatogram showing solute concentration as a function of time.

Figure 1: Schematic of a Gas Chromatograph

Ideally, a component’s peak height should depend on its concentration but in some instances, the
peaks do not have an ideal (symmetric) shape. Peak area then becomes a more accurate measure

Prepared by: K.K.R. – July 2017

of concentration. The retention time refers to the time period between injection and detection of a
particular component. It is used to identify the components present in the sample. Components
that strongly interact with the stationary phase have a longer duration within the column and thus
have a greater retention time.

A calibration by using an internal or external standard may be employed. One difference is that a
known quantity of the internal standard is added to all calibration standards and samples whilst
an external standard is analysed separately under identical conditions. The calibration when
using an internal standard is based on the ratio of the response of the analyte to the internal
standard. An internal standard allows for correction of analyte losses during preparation which is
not accounted for when using an external standard.

To construct a calibration curve, a given amount of the internal standard solution is added to a
reference standard. The area ratio of the reference to the internal standard is then plotted against
the ethanol standard concentrations to produce the calibration curve. To prepare the ethanol
standards, equation (1) is used and the area ratio is determined using equation (2).

(1)
Where:
- Concentration and volume of the standard solution
- Concentration and volume of the pure ethanol

To determine the concentration of the analyte, the same procedure is followed but the reference
standard is replaced by the analyte. The peak area ratio of the analyte to the internal standard can
then be used to find the unknown concentration using the equation below:

Where:
c – y-intercept of the calibration curve
m – Slope of the line

Prepared by: K.K.R. – July 2017

APPARATUS AND REAGENTS

 n-propanol  Volumetric flasks (10ml (10), 50ml

 99.8% ethanol (6), 100ml (1))
 4 brands of beer (Carib Pilsner,  12 beakers
Heineken, Heineken Light, Stag)  2 funnels
 Distilled water  No. 40 filter paper
 Agilent Technologies 7890A Gas  Crimper
Chromatograph System  De-crimper
 10 2ml vials with covers  Droppers
 Pipettes (1ml, 2.5ml, 5ml) and tips  Funnel rack

PROCEDURE

A. Preparation and analysis of Calibration Standards

1. Prepare internal standard, 5% n-propanol stock solution by adding 5ml of n-propanol to a

100ml volumetric flask and fill up to the calibration mark with distilled water.

2. Prepare a series of ethanol standard solutions of 2, 3, 4, 5, 6 and 7% in 50ml. Using equation

(1), complete the table below.

Table 1: Volumes to be used when preparing ethanol standard solutions

Concentration of Volume of Concentration Volume of pure Volume of distilled

standard (%) standard (ml) of ethanol (%) ethanol (ml) water (ml)
2
3
4
50 100
5
6
7

Prepared by: K.K.R. – July 2017

3. Pipet 5ml of each ethanol standard solution into a separate volumetric flask and label all
flasks.
4. Pipet 5ml internal standard solution into each flask and mix well. Ensure that new pipette tips
are used for different solutions.
5. Fill 2ml vials with each standard solution, cover tightly using the crimper, label and load into
the sampler tray.
6. Ensure that the system is set for double injection, start the sequence and record the results in
terms of peak area.

B. Analysis of samples
1. Record the alcohol content stated on each bottle. Decarbonate each beer sample by filtering
through No. 40 filter paper.
2. Pipet 5ml of each sample into 10ml volumetric flasks, label and add 5ml aqueous n-propanol
standard solution.
3. Mix thoroughly by swirling.
4. Fill 2ml vials with each mixture, cover tightly using the crimper, label and load into sampler
tray.
5. Ensure that the system is set for double injection, start the sequence and record the results in
terms of peak area.

RESULTS AND CALCULATIONS

1. Determine the uncertainty in measurements of the three pipettes used and briefly explain this
concept.
2. Tabulate all results and calculate the mean and standard deviation using the equation below:

Where:
σ – Standard deviation

Prepared by: K.K.R. – July 2017

x – Each value in the data set
– Mean of all values in the data set
n – Number of values in the data set

3. Using the average peak areas, calculate the area ratio of the ethanol to n-propanol peaks for:
i. Calibration standard solutions (Procedure A)
ii. Beer samples (Procedure B)
4. Construct the calibration curve by plotting area ratio of ethanol/n-propanol (from (i) above)
against ethanol standard concentrations.
5. From the equation of the line, state the slope (m) and y-intercept (c) and include the R2
value.
6. Determine the concentration of ethanol in the various beer samples using equation (3).
7. Compare and contrast results obtained with the alcohol content stated on the labels. Account
for differences between the calculated and stated alcohol percent.

Note:

In the executive summary:

- Comment on the suitability of the n-propanol standard solution as the internal standard.
- Discuss the reason(s) as to why ethanol was eluted before n-propanol.
- State the sources of error which would account for discrepancies in the results and
propose suitable recommendations.

Prepared by: K.K.R. – July 2017