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Basic gas chromatography

prof. aza
Department of Pharmacy,
Andalas University
STIFI Perintis, Padang
STIFAR, Pekan Baru
STIFI Bhakti Pertiwi, Palembang

prof. aza
Definition
• The basis for gas chromatographic separation is
the distribution of a sample between two phases.
One of these phases is stationary bed of large
surface area, and the other phase is gas which
percolates through the stationary bed.
• If the stationary phase is a solid, we speak of Gas-
Solid Chromatography. This depends up on the
adsorptive properties of the column packing to
separate samples, primarily gases.
• If the stationary phase is a liquid, we speak of Gas
liquid Chromatography (GLC). The liquid is spread
as a thin film over an inert solid and the basis for
separation is the partitioning of the sample in and
out of this liquid film.
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Schematic drawing of Gas Chromatographic System

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GC/MS • Gas Chromatography/Mass
Spectrometry (GC/MS) is a
combination of two
instrumental techniques,
gas chromatography and
mass spectrometry. The
gas chromatograph is used
to separate a mixture into
component parts and
deliver them to the mass
spectrometer. The mass
spectrometer breaks the
molecules into ions and
records the resulting
spectrum. This spectrum
conclusively identifies the
compound.

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Chromatogram,
A Typical gas-
liquid
chromatography
displays each
component as
peak

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Resolution
• The resolution of chromatographic peaks is
related to two factors: column efficiency, and
solvent efficiency.
• Column efficiency is concerned with the peak
broadening of initially compact band as it passes
through the column.
• The broadening results from the column design
and operating conditions, and can be quantitatively
described by the height equivalent to a
theoretical plate.
• The HETP is that length of column necessary for
the attainment of solute equilibrium between the
moving gas phase and stationary liquid phase.
prof. aza
Hypothetical representation of
Chromatographic separation
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Advantages of Gas
Chromatography
• Speed, the entire analysis is completed within 23
minutes.
• High Resolution. By using selective solvents, GC
can provide resolution impossible by distillation or
other techniques.
• Qualitative analysis. The retention time is that
time from injection to the peak maxima. This
property is characteristic of the sample and the
liquid phase at a given temperature. Each
compound has only one retention time. This
retention time is not influenced by the presence
of other component.

prof. aza
Advantages of Gas Chromatography,
continued
• Quantitative analysis. The area produced for each
peak is proportional to that peak’s concentration.
This can be used to determine the exact
concentration for each component.
• Sensitivity. The simplest forms of thermal
conductivity cells can determine down 0.01 % (100
ppm). The flame detector easily sees parts per
million, and the specific electron capture detector
can measure parts per billion or picograms (10-12 g).
• Simplicity. GC are simple to operate and
understand. Interpretation of the data obtained is
usually rapid and straight forward.

prof. aza
Temperature
• Injection-port temperature should be hot enough
to vaporize the sample so rapidly that no loss in
efficiency results from the injection technique
and must be low enough so that thermal
decomposition or rearrangement is avoided.
• Column temperature should be high enough so that
the analysis is accomplished in a reasonable length
of time. The retention time doubles for every 300
C decrease in column temperature. For the most
samples the lower the column operating
temperature, the higher the ratio of partition
coefficient in the stationary phase and the better
the resultant separation.
• Detector temperature should be high enough so
that condensation of the sample and or liquid
phase does not occur.
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Detector Sensitivity

TCD All 10 ppm


compound
FID Organic 0.1 ppm
substances
ECD 0.1 ppb

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Thermal conductivity Detector
(TCD)
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Flame ionization Detector
(FID)

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Electron capture detector
(ECD)
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Carrier gas
• Commonly used gases are hydrogen,
helium, und nitrogen. The carrier gas
should be :
• Inert to avoid interaction with sample
or solvent.
• Able to minimize gaseous diffusion.
• Inexpensive.
• Suitable for detector used.
prof. aza
Solid support
The purpose of the solid support is to
provide a large uniform, inert surface area
for distributing the liquid phase. Some
desirable support properties are :
• Inert (avoid adsorption).
• High crushing strength.
• Large surface area.
• Regulator shape, uniform size.

prof. aza
Stationary phase
• Ideally the solvent should have the
following characteristic:
• Samples must exhibit different
distribution coefficients.
• Sample should have a reasonable
solubility in the solvent.
• Solvent should have a negligible vapor
pressure at operating temperature.
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Resolution
• The resolution of chromatographic peaks is
related to two factors: column efficiency, and
solvent efficiency.
• Column efficiency is concerned with the peak
broadening of initially compact band as it passes
through the column.
• The broadening results from the column design
and operating conditions, and can be quantitatively
described by the height equivalent to a
theoretical plate.
• The HETP is that length of column necessary for
the attainment of solute equilibrium between the
moving gas phase and stationary liquid phase.
prof. aza
Resolution, continued
• Solvent efficiency result from the solute-
solvent interaction and determines the
relative position of solute bands on a
chromatogram.
• Solvent efficiency is expressed as the
ratio of peak maxima (adjusted retention
times).
• Column efficiency is measured by the
number of the theoretical plates.
prof. aza
Resolution, continued

The true separation of two


consecutive peaks is
measured by the resolution.
Resolution is a measure of
both the column and solvent
efficiency.
If R = 1, the resolution of
two equal-area peaks is
approximately 98% complex.
If R = 1,5, base line
separation (99,7% resolution)
ia achieved.
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Separation of
two bands as
a function of
resolution
(Rs) and
relative band
size (I/1, ¼,
1/16).

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Solvent efficiency and Separation
Factor

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Normal

Increased Column Efficiency


(more theoretical plate)

Increased Solvent Efficiency


(greater ratio of retention
times) prof. aza
Effects of a
change in k’,
N, or a on the
resolution of
two bands

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Calculation of Theoretical Plate

HETP = L/N
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Band asymmetry,
(a) Definition of
asymmetry factor,
As; (b-e) examples
of band asymmetry

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Rate theory, van Deemter
• The three principal contributions to
the broadening of a band are :
• Multipath effect or eddy diffusion (A
term)
• Molecular diffusion (B term).
• Resistance to mass transfer (gas and
liquid, C term).

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Plot of HETP against gas velocity

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van Deemter equation

HETP = A + B/µ + C.µ

µ = length of column (cm)/ retention time (seconds)


HETP = 2 dp + 2 Dgas + 8 k’ df 2 x µ
µ  (1+k’)2 Dliq

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van Deemter equation, continued

• 2 dp • 2 Dgas
 = a constant which is a µ
measures of packing Dgas = diffusivity of the
irregularities. solute in the gas phase.
dp = average particle µ = the linear gas
diameter of the solid velocity
support.

prof. aza
van Deemter equation, continued

• 8 k’ df2 • Fliq = fraction of cross


(1+k’)2 Dliq section occupied by the
liquid phase
k’ = capacity factor =
• Fgas = fraction of cross
k ( Fliq / Fgas ) section occupied by the
k = partition coefficient of gas phase
the solute, expressed as • df = effective thickness of
the amount of solute per the liquid film which is
coated on the particles of
unit volume of liquid
the support.
phase divided by the
• Dliq = diffusivity of solute
amount of solute per unit
in the liquid phase
volume of gas phase

prof. aza
Illustration of Multipaths

Multiple Path (Eddy diffusion) term, A = 2 dp


In any packed column solute molecules and carrier gas
molecules travel along many paths. These path have
different lengths, therefore the solute molecules have
different residence time. This adds to peak broadening.
This broadening depends upon the size of the particle
constituting the packing, the shape, and the manner in
which they are packed and the column diameter.

prof. aza
To improve column efficiency
• Particle diameter: Column efficiency is improved
by the use of small, uniform particle size.
Diatomaceous earth type with 100 – 120 mesh
range.
• Flow rate: For maximum efficiency the column
must be operated at the optimum flow rate. In
practice, operating at flow rates slightly higher
than optimum will decrease the analysis time and
not materially effect the HETP.
• Carrier gas: The detector employed usually
dictates the choice of carrier gas. For highest
efficiency a high molecular weight gas should be
choice. Where rapid analysis time is required and
highest efficiency is not necessary a low molecular
weight carrier gas such as helium or hydrogen
would be preferred.
prof. aza
To improve column efficiency, continued
• Type of liquid phase: A low viscosity,
low vapor pressures solvent with good
absolute solubility for the sample
should be used. To obtain a
separation, it must exhibit a
differential solubility.
• Amount of liquid phase: Low liquid
loadings (thin film) 1% - 10% have the
advantage of fast analysis and lower
temperature operation.
prof. aza
To improve column efficiency, continued
• Temperature: Resolution can usually be improved
by lowering the column temperature. Lowering the
temperature also decreases decomposition of the
compounds but may increase adsorption.
• Pressure: The majority of practical gas
chromatographers work at an outlet pressure of
one atmosphere so that operation at the optimum
flow rate fixes the inlet pressure. Best efficiency
is obtained at low inlet-to-outlet pressure ratios.
• Column diameter: Capillary and preparative column
experiments indicate that efficiency is improved
with decreasing internal diameter. Thus 1/8 – 1/16
inch OD are used for highest resolution.

prof. aza
Basic measurement

prof. aza
Science for a
better future

1955 – 2005 FMIPA UNAND


prof. aza