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Physical chemistry lab courses for beginners

GC
Experiment A14: Basics of gas chromatography

Important: Please bring a USB stick!

1. Introduction

Along with distillation and extraction, gas chromatography is one of the most important thermal
separation processes and represents a sensitive method for identifying and analyzing the
components of mixtures of substances. Gas chromatography is used, among other things, in the
analysis of agricultural and meat products, in the analysis of pharmaceuticals, petroleum
components and in environmental analysis. The enormous importance of chromatography in
chemistry was made clear by the award of the 1952 Nobel Prize in Chemistry to two pioneers
of partitions chromatography, Archer J. P. Martin and Richard L.M. Synge.

2. Theoretical background

The separation of individual components of a mixture takes place in a separation section

(column) consisting of a stationary phase. The mixture as a component of a mobile phase
(mixture in carrier gas) flows through the stationary phase. The interaction of the molecules of
the mobile phase with the components of the column is molecule-specific and leads to different
flow rates of the transported components. The molecules of the mixture oscillate back and forth
between the phases and thus exhibit different residence times in the stationary phase. The
extended residence times are characterized by thermodynamic equilibria between the gas and
solid phases (adsorption/desorption) and thus depend on the temperature of the gas/solid
interface. This fact is expressed by measurably different delays in the transport of the individual
components of the mixture. This delay is called chromatographic retention. The different
retention times in the stationary phase are determined by the free Gibbs enthalpy of the delay
process (adsorption/desorption). The time required for the molecules to cross the separation
distance is called residence time. Two components of a mixture are therefore separable if they
have different gross retention times, which conversely means that they differ in their interaction
with the stationary phase. Their separability is described by the so-called separation factor ∆ (∆
= t1/t2, the ratio of the retention times of the two components). This factor reflects the ratios of
all derived variables, e.g. quantity ratios, distribution coefficients, etc..

The retention time of a component is measured by means of a detector positioned at the exit of
the column. The gas flowing past changes the temperature of the detector, which can be detected
by the change in resistance. Due to the statistical nature of molecular transport (diffusion
processes lead to different path lengths of the molecules) the measurement signal of a
transported component shows an approximate Gaussian shape. The retention time tR, the time
between injection of the component until the Gauss peak (maximum) appears on the detector,
remains constant under the same chromatographic conditions and is therefore characteristic of
the component under investigation. The identification of an unknown component can therefore
be done by comparing the retention times with a standard.
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measured data
Baseline

Intensity I

t=0 t0 tR1 tR2 tR3 tR4

Time t

The retention volume VR (the amount of mobile phase that has flowed through the separation
system in time tR) can be calculated from the following relation:

1) VR = tR F, where F - constant flowrate (mL/min).

A comparison of the absolute retention times is difficult because tR depends on the flow
velocity of the mobile phase as well as on the length L of the separation distance. The so-called
capacity factor (k* value) is a quantity independent of F and L. This quantity relates the
retention time tR* to the dead time t0:

2) k*= tR*/t0 = (tR-t0)/t0 = (tR/t0) -1

Thus, The k* value represents the molar ratio of a component in stationary and mobile phase.
The k* value is linked to the distribution coefficient K:

3) k* = (Vs/Vg)K

where Vs is the volume oft he stationary phase and Vg is the volume of the mobile phase. The
capacity factor k* is thus proportional to the volume of the stationary phase (or the specific
surface area of the adsorbates, inm2/g ).

The relative retention α for two components 1 and 2 can thus be represented as follows:

4) α= k2*/ k1* = K2 / K1,

i.e. two components with identical k* values remain inseparable.

In contrast to distillation and extraction, chromatography does not have any physical separation
stages; in simple terms, the process is modelled as follows: a certain amount of the gas mixture
enters a section of the column at a certain point in time, which is regarded as the theoretical
separation stage. In this section each gas molecule is adsorbed with a substance-specific
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probability (red). All non-adsorbed molecules (blue) move to the next separation stage in the
next time step, all adsorbed molecules remain. Now all gas molecules can be adsorbed again
with a substance-specific probability, while all adsorbate molecules desorb with a substance-
specific probability. Again all non-adsorbed molecules move one separation stage further, while
all adsorbed molecules remain in their respective stage. This is repeated until all molecules have
passed all separation stages. As long as the column is not overloaded by too large sample
volumes, incorrectly selected temperature or flow rate, the separation is statistical and the
individual components leave the column Gaussian-distributed. Otherwise the measured peak
will be deformed.

According to the theory of Martin and Synge from 1941, the resolving power of a column of
length L can be described with the parameter HETP (height equivalent to a theoretical plate):

5) HETP = L / N (in mm)

where N, the number of separation stages in the column (plate number), can be calculated for
Gaussian peaks as follows:

6) N = 16 (tR / W)2

Where tR is the residence time at the peak maximum and W is the peak base width (distance of
the intersection of the turning tangents with the base line). For peaks that deviate from the
Gaussian shape, the base width W is replaced by the half-width W1/2:

7) N = 8 ln 2 (tR / W1/2)2 therefore HETP= L /[8ln2 (tR / W1/2)2]

The resolution R of a GC separation path is defined by the ratio of the retention time difference
of the adjacent peaks ∆tR, and the arithmetic mean of the corresponding base widths W:

8) R = (tR2 - tR1) / (W(1)+ W(2))/2 = 1,198 (tR2 - tR1) / [W1/2(1)+ W1/2(2)]

The linear flow rate u of the mobile phase is calculated from the chromatogram as follows

9) u = Lg / t0
Lg Length of the separation column incl. inlet and outlet in cm
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t0 Dead time of the column in s at this flow rate

If the dead time signal is not visible in the chromatogram t0 can be calculated approximately in
this way:

10) t0=A*Lg/ F ( u=F/A)

A= Cross sectional area of the column tube
F= Flow rate according to flow meter

According to Van Deemter, the HETP parameter depends on the flow of the mobile phase u
This equation describes chromatographic separation as a dynamic process and takes into
account the role of diffusion and mass transfer:

11) HETP = A + B/u + C u.

A - Contribution of the Eddy diffusion (diffusion caused by turbulences)

B - Contribution of longitudinal diffusion
C=[ K1df2/DS + K2dP2/DM] Mass transfer from and to stationary phase.
u – Linear flow rate
λ - Constant, that describes particle shape and homogeneity oft he package.
dP - Particle diameter
DM - Diffusion coefficient of the mobile phase
Ψ – Correction factor for the free spaces between the particles
df - Thickness of the liquid film
K1, K2: Correction factors for the specific geometry of the column and the column capacity.

The eddy diffusion A is only dependent on the packing of the stationary phase and the
associated path that the mobile phase must travel in it. The smaller and more homogeneously
packed the column, the smaller the eddy diffusion. It is independent of the flow velocity u.
A=2*λ*dP
The longitudinal diffusion B is only dependent on the mobile phase. It occurs in both packed
and unpacked columns and describes the widening of the phase due to diffusion away from the
center. The greater u the smaller the contribution of B. B=2*λ∗ DM.
C is the mass transfer between stationary and mobile phase and thus describes the actual
chromatographic process. C differs from particle to particle and thus enables the separation of
different components of the mobile phase. The slower the flow velocity, the more effective is
the separation of the individual components.C=[ K1df2/DS + K2dP2/DM]

This equation shows how to find the optimum separation performance of the column (optimum:
shortest possible analysis time and good separation performance). By analyzing the specific
parameters (λ, dP, dM …) it is possible to make statements about the optimal column
composition: columns with a small diameter, which are tightly and homogeneously packed with
fine-grained material, show the best separation performance.
At low flow rates u the role of the A and C terms can be neglected and eq. 9 allows to determine
the effective coefficient of longitudinal diffusion (eq.11):

12) HETP = 2 Ψ DM /u ,

whereby in practice the parameter Ψ≈1 is set,

13) Deff = HETP u /2.