Unit 3 Gas Chromatography

UNIT 3
GAS CHROMATOGRAPHY

Structure
3.1 Introduction Sample into GC Unit
Expected Learning Outcomes Hazards in Sampling
3.2 Recapitulation of Basic Aspects Introduction of the Sample into GC
Unit
Column Efficiency
3.5 Applications
Solvent Efficiency
Identification of Compounds
3.3 Instrumentation
(Qualitative Analysis)
Mobile Phase (Carrier Gas)
Quantitative Analysis
Columns
Some Examples of Applications
Stationary Phase Support and
3.6 Summary
Liquid Phases
3.7 Terminal Questions
Detectors
3.8 Answers
3.4 Sampling and Introduction of

3.1 INTRODUCTION
In the earlier units (i.e. Units 4 and 5), it was emphasized that a reasonable
classification of chromatographic techniques can be proposed on the basis of
the nature of the mobile phase. Liquid chromatography, where the mobile
phase is liquid, has several diversifications depending upon the shape of
support, nature of support and the mechanism responsible for separation. If
gas is the mobile phase, the chromatographic technique is known as gas
chromatography (GC). It is used for the separation of gaseous or volatile
substances. It has only two sub-classifications namely gas-solid
chromatography (GSC) and gas- liquid chromatography (GLC). However, in
terms of importance and applications, it is not second to any chromatographic
technique. It may be important to point out here that the theoretical aspects of
chromatographic techniques have been developed taking gas chromatography
as the model.

In gas chromatography, the basis for separation is the distribution of solutes

between two phases. One of these phases is a stationary bed of large surface
area (stationary phase) and the other is a gas (mobile phase) which
percolates through the stationary phase. If the stationary phase is a solid
93
Block 1 Solvent Extraction and Chromatographic Methods

adsorbent, then it is known as gas- solid chromatography (GSC). Here, the

adsorptive properties of the stationary phase are responsible for separating
solutes, primarily gases. Common solid stationary phases are silica gel,
molecular sieves, porous polymers, alumina and charcoal. In case, the
stationary phase is liquid, it is called gas- liquid chromatography(GLC). The
liquid is spread (coated) as thin film over an inert support. The basis of
separation is the partitioning of the solutes in and out of the liquid film. There
is a wide range of the liquid phases with usuable temperatures up to 400ºC.
This makes GLC the most versatile and the selective form of chromatography.
It is used for analysis of gases, liquids and solids.

This particular unit discusses the both theoretical and practical aspects of gas
chromatography. The theory of chromatography has been explained in Unit 4
where there was an emphasis on gas as a mobile phase. Here, we will
recapitulate the basic concepts with a total focus on gas chromatography.
Some of the symbols used here may be different from those used in Unit 4 but
they have been properly explained to avoid any confusion. Under the basic
aspects, we will discuss the working of a gas chromatograph and the column
and solvent efficiency. This is followed by a discussion on instrumentation
including requirements of a carrier gas. This is followed by a discussion on
columns, stationary phase support and liquid phases and finally the detectors.
The next section describes sampling and sample injection. The last section of
the unit illustrates some representative applications in terms of both qualitative
and quantitative analysis.

Expected Learning Outcomes

After study this Unit, you should be able to:
 discuss the basic aspects of gas chromatography particularly in terms of
column and solvent efficiency;
 explain the importance of characteristics of carrier gas;
 describe about different types of columns;
 select appropriate stationary phase support and liquid phase;
 explain the working of different types of detectors and select the
appropriate one;
 discuss the importance of sampling and sample injection in GC unit; and
 describe the utility of the technique for both qualitative and quantitative
analysis.

3.2 RECAPITULATION OF BASIC ASPECTS

In the operation of a gas chromatograph the, solutes in a mixture are
completely vaporised in the injection port and they are moved through the
column by a carrier gas under pressure. It is in the column where separation
takes place. From the column, the separated solutes pass through a detector
where they are sensed generating an electronic signal. The signal is then
amplified and normally displayed on a strip chart recorder. The trace plotted
94 on the recorder is called a "Chromatogram". It is a plot of the detector
Unit 3 Gas Chromatography

response in minivolts as a function of time. Usually, time is the abscissa and

minivolts the ordinate.

Fig. 3.1: A typical chromatogram.

From the chromatogram, several general observations can be made.

Under a given set of experimental conditions, each peak has a characteristic

retention time (tR) and the retention volume (VR) that are useful in qualitative
analysis of solutes. The retention time for solute A is depicted in the figure as
the distance from the point of injection to the peak maximum (tR)A. Similarly,
(tR)B and (tR)C are the retention times for solutes B and C, respectively. If the
flow rate of carrier gas, Fc is constant, then

V R = tR × F c … (3.1)

where, VR is the retention volume. Thus, (VR)A, (VR)B, and (VR)C are the
retention volumes of solutes A,B and C, respectively. In practice, one can use
either retention time (tR) or retention volume (VR) for qualitative analysis [see
Fig 3.2 (a) and (b)].

(b)
Fig. 3.2 (a): Chromatogram of sample containing 'A' and 'C’; (b) Chromatogram
of standard solution containing ‘A’, ‘B’ and ‘C’. 95
Block 1 Solvent Extraction and Chromatographic Methods

In this Unit, we shall restrict to retention volume, VR, for our discussion. The
small early peak in Fig. 3.1 represents a solute which is least retained by the
stationary phase. It passes straight through the column without stopping. In
GC, this behaviour is often shown by air or methane and the peak is often
called an air peak. The symbol Vm, sometimes called the hold-up volume or
void volume, serves to measure the interstitial or interparticle volume of the
column. The term dead volume, while not recommended, is also widely used.

tR  tR  t0 ... (3.2)

where tR = adjusted retention time and t0 = retention time of air peak Thus, (tR)
A,

(tR) B and (tR) C are the adjusted retention times for solutes A, B, and C,
respectively.

Similarly

VR/=VR–VM ... (3.3)

where V R = adjusted retention volume and VM = hold up volume and ( VR ) A,

( VR ) B and ( VR ) C are the adjusted retention volumes for solutes A, B, and C,
respectively. The detector response, or area under the curve, (AUC) is
proportional to the quantity of material passing through the column. Therefore,
a quantitative measurement can be made. For this purpose, a calibration
graph of AUC vs quantity of solute injected need to be constructed for each
solute separately. If we refer to Fig. 3.1, based on AUC, solute B is present in
the largest amount, while A and C are in lesser amounts and are
approximately equal.

The main interest of any chromatographer is good separation or resolution

between neighbouring peaks. This helps in accurate qualitative analysis and
quantitative determination of solutes being separated from a mixture. The
resolution of chromatographic peaks is related to the following two factors:

1. Column efficiency

2. Separation factor ( Solvent efficiency)

The column efficiency is concerned with the peak broadening of an initially

compact band as it passes through the column. The broadening results from
the column design and operating conditions. This can be quantitatively
described by the plate height (H), which is that length of the column necessary
for the attainment of solute equilibrium between the moving gas phase and
the stationary liquid phase. The separation factor, (α), on the other hand,
results from the solute-solvent (liquid phase) interaction and determines the
relative position of solute peaks on a chromatogram. The separation factors or
relative retention is expressed as the ratio of retention times. Thus α is
determined by the respective distribution constants (Kc) of the solutes in the
liquid phase at a given temperature. The idea of increased column and
96 solvent efficiency is illustrated in Fig 3.3.
Unit 3 Gas Chromatography

Fig. 3.3: Illustration of column and solvent efficiency.

3.2.1 Column Efficiency

Column .efficiency is measured by the plate number, N. To compare column
efficiency, one must specify the liquid stationary phase, solute (s), temperature, flow
rate of carrier gas and sample size. As stated in Unit 4, the plate concept is a carry
over from distillation process where historically, the first efficient columns were in
fact composed of discrete plates. In the laboratory, however, packed distillation
columns are more commonly employed and the plate number is a theoretical
concept introduced so that column performance can be evaluated.

In Gas Chromatography, as in distillation, the discrete plate is an artificial concept.

The plate number obtained in the methods varies greatly and should not be taken
as a direct measure of the difference in their separating power. Plate number is
useful to compare similar columns, or set standards for packing techniques. The
plate number can be easily measured from the chromatograms. Fig. 3.4
shows the measurements needed to make this calculation.

Fig. 3.4: A normal distribution curve. The inflection point at 0.607 of the peak
height. The quantity w b is the width at 0.005 of the peak height (half-
height) and corresponds to 2.354 σ. The quantity w b is the base width
and corresponds to 4σ as indicated.
97
Block 1 Solvent Extraction and Chromatographic Methods

tR 2 t t
N= ( ) = 16( R ) 2 = 5.54( R ) 2 … (3.4)
σ wb wb
Different terms arise because the measurement of  can be made at different
heights in the peak. At the base of the peak wb is 4, so the numerical
constant is 4 2 or 16. At half height ,wh 2.354σ and the constant becomes 5.54.
Independent of the symbols used, both the numerator and denominator must
be given in the same units and, therefore, N is unitless. Typically, both the
retention time and the peak width are measured as distance on the
chromatographic chart. Alternatively, both could be in either volume units or
time units. No matter which calculation is made, a large value for N indicates
an efficient column which is highly desirable. A related parameter which
expresses the efficiency of a column is the plate height, (H)

wH = L/N … (3.5)

where, L is the column length, H has the units of length and is better than N
for comparing efficiencies of columns of differing lengths. It is also called the
Height Equivalent to one Theoretical Plate (HETP), a term which carried over
from distillation terminology. A good column will have a large N and small H.

Another measure of the efficiency of a column is resolution, Rs. As in other

analytical techniques, the term resolution is used to express the degree to
which adjacent peaks are separated. For chromatography, the definition is,
(t R ) B  (t R ) A 2d
Rs = = … (3.6)
( wb ) A  ( wb ) B ( wb ) A  ( wb ) B
2

where, d is the distance between the peak maxima for two solutes A and B.
Fig. 3.5 illustrates the way in which resolution is calculated. Tangents axe
drawn to the inflection points in order to determine the widths of the peaks at
their bases. Normally, adjacent peaks of equal area will have equal the same
peak widths, and (wb)A will equal (wb )B. Therefore, Eq. 3.6 is reduced to the
following:
d
Rs = … (3.7)
wb

In Fig. 3.5, the tangents are touching so d = wb and Rs = 1.0. The larger the
value of resolution, the better the separation complete baseline separation
requires a resolution of 1.5.

98 Fig. 3.5: Two nearly resolving peaks illustrating the definition of resolution, Rs
Unit 3 Gas Chromatography

Rate Theory

There has to be quantitative relationship between plate height and column

variables. In this context, several chromatographic theories have been
developed to account for the shape of eluting curves from chromatographic
columns. The rate theory was developed by van Deemter. A qualitative
understanding of the van Deemter equation is useful in optimizing the
chromatographic performance.

The three principal contributions to the broadening of a band are :

1. Multiple path effect or eddy diffusion (A term)

2. Molecular diffusion (B term)

3. Resistance to mass transfer (gas and liquid, C term)

From these, a basic equation can be derived for the plate height in a gas liquid
system.
H= A+B/u+Cu … (3.8)

where A, B, and C are above mentioned constants and u the linear velocity (or
flow rate) through the chromatographic column. The linear gas velocity is
measured by
Len gth of c o lu m n (cm )
u 
Rete ntion tim e of air (seco nds )
If H is plotted against u, one obtains a hyperbola with a minimum H. This
minimum is that flow rate (u optimum) at which the column is operating most
efficiently. However, owing to compressibility of the carrier gas, u is not
constant over the entire length of the column; hence, only a small section can
operate at maximum efficiency. Here, the van Deemter equation is being
presented in an expanded form. The significance of the terms is the same as
Unit 4.

The expanded version of the van Deemter equation is:

2Dgas 8k d 2 f
HETP = 2λdp + + 2
u … (3.9)
u  2 (1  k ') Dliq
where,

 = a parameter which is a measure of packing irregularities,

dp= diameter of packing particles,

 = a correction factor accounting for the tortuosity of the gas channels in the
column,

df = effective film thickness, a measure of liquid loading of packing material,

Dliq = diffusion coefficient of the solute in the liquid phase,

Dgas = diffusivity of the solute in the gas phase,

u = the linear gas velocity, and

k´= capacity factor = k (Fliq/Fgas). 99

Block 1 Solvent Extraction and Chromatographic Methods

Fig. 3.6: A typical van Deemter plot showing the effect of eddy diffusion (A),
longitudinal diffusion (B/u) and mass transfer for both phases (Cu).

The conclusions which have been drawn from the rate theory of van Deemter
are of practical interest and can be used to improve column efficiency.

i) Particle diameter: Column efficiency is improved by the use of small

and uniform particle size. Supports which give the highest efficiency are
diatomaceous earth type with 100 - 120 mesh range.

ii) Flow rate : For maximum efficiency, the column must be operated at the
optimum flow rate. This is found by a plot of H against the flow rate. The
minimum H determines the optimum linear gas velocity or a flow rate to
be used. In practice, operating at flow rates slightly higher than optimum
will decrease the analysis time and not materially affect the H.

iii) Carrier gas : The detector employed usually dictates the choice of
carrier gas. However for the high efficiency, a high molecular weight gas
should be the choice. Where rapid analysis time is required and high
efficiency is not necessary, a low molecular weight carrier gas such as
helium or hydrogen would be preferred.

iv) Type of liquid phase : A low viscosity, low vapor pressure solvent with
good absolute solubility for the sample should be used. To obtain a
separation, it must also exhibit a differential solubility.

v) Amount of liquid phase : Low liquid loadings (thin film ) 1 % - 10 %

have the advantage of fast analysis and lower temperature operation.
Low liquid loadings, however, reduce the sample capacity and may
require highly inactive solid supports.

vi) Pressure: A majority of practical gas chromatographers work at an outlet

pressure of one atmosphere so that operation at optimum flow rate fixes
the inlet pressure. However, best efficiency is obtained at low inlet-to-
100 outlet pressure ratios.
Unit 3 Gas Chromatography

vii) Temperature: The resolution can usually be improved by lowering the

column temperature. The practical problem with low temperature results
in long analysis time. Lowering the temperatures also decreases
decomposition of the compounds but may increase adsorption.
Simultaneous reduction of liquid loading and temperature is generally
beneficial. The limit to this is the increased exposure of active sites on
the solid support and the need for sensitive detector.

viii) Column diameter : Capillary and preparative column experiments

indicate that efficiency is improved with decreasing internal diameter.

3.2.2 Solvent Efficiency

A striking feature of gas chromatography is that substances having the same
vapor pressure can be easily separated by appropriate selections of the liquid
phase. The major advantage of GC over distillation is the ability to use
selective solvents. There are numerous selective phases available and the
following factors should aid in selection of a proper one.

1. Interaction forces and partition coefficient:

There are following four interaction forces which can aid in the GC
separation:

a) Orientation or Keesom forces : Forces resulting .from the

interaction between two permanent dipoles. The “ hydrogen-bond”
is a particularly important type of orientation force encountered in
gas chromatography.

b) Induced dipole or Debye forces : Forces resulting from the

interaction between a permanent dipole in one molecule and the
induced dipole in a neighbouring molecule. These forces are
usually very small.

c) Dispersion, London or non-polar forces : Forces arising from

synchronized variations in the instantaneous dipoles of the two
interacting species . These forces are present in all cases and are
the only source of attraction energy between two non-polar
substances. They are weak compared to (a) and (b).

d) Specific interaction forces : Forces resulting from chemical bonding

, complex formation between solute and solvents . These forces of
interaction determine the solubility and thereby, the separation is
achieved. Their combined effects are expressed by the distribution
constant Kc, where

A m oun t of so lu te pe r un it v o lu m e of liquid p ha se
Kc 
A m ou n t of so lut e pe r un it v o lu m e o f g as p ha se
… (3.10)

The value of Kc is high when most of a solute is retained in the

liquid phase. This means that the solute moves slowly down the
column because only a small fraction will be in the carrier gas at
any given time. Transport is negligible in the liquid phase and only 101
Block 1 Solvent Extraction and Chromatographic Methods

that fraction in the gas phase is carried through the column. Thus,
separation between two solutes is possible if their distribution
constants are dissimilar . The greater the difference in their Kc
values, the fewer the plates or the shorter the column length that is
required to achieve a separation.

2. Solvent efficiency and temperature

Solvent efficiency is measured by α, the relative retention. It is the ratio

of adjusted retention times or partition coefficients. Calculation of solvent
efficiency is illustrated in Fig. 3.7.

Fig. 3.7: Calculation of solvent efficiency.

Relative retention differs from the separation factor (S.F.), where S.F. is
the ratio of uncorrected retention times. Both α and Kc are temperature
dependent. However, over a limited temperature range, α ill be
constant. The distribution constant, decreases with increasing
temperature, i.e., the fraction of the solute in the gas •phase will increase
and hence, the elution time will decrease. This results in decreased
separation since it is the liquid phase which performs the separation. No
separation occurs in the gas phase. To achieve better separations, lower
temperatures should be used. Lower temperatures mean more liquid
phase interaction, more separation and longer analysis time. At a
minimum, the solute should spend 50 % of the time in the liquid phase
so that the retention time exceeds twice the retention time of air.

3. Number of plates for required separation

An equation which is useful in determining the number of plates and,

thus, the length of column required is given below:
α 2 k2  1 2
Nreq= 16 Rs2 ( α  1) ( k ) … (3.11)
2

where, k´2= capacity factor for peak 2

A djust ed re ten t io n tim e , X s

 … (3.12)
Ree n tion t im e a ir

where, Rs is the resolution required, and α is the solvent efficiency, both

as defined above. The number of plates required can be illustrated by
102 the following example.
Unit 3 Gas Chromatography

Example

The separation shown below was obtained with a 3 m column. What is the
minimum column length necessary to obtain a resolution of 1.5?

Fig. 3.8: Calculation of required plates

2 2
x  17 
Norig  16    1 6    46 24 p la tes
y  1 
17  1 17  1
  1 .2 3 1; k2   16
14  1 1

2 2
    k2  1 
2
Nreq  16 R    
   1   k2 
2 2
 1.2 31   1 6  1 
 16(1 .5)2    
 1 .23 1  1   16 
 36  2 8.4  1.13   11 55 p la t es
Nreq
Lreq d  Lo rig 
N orig

1 155
 3  0.75 m = 0.75 m
4 624

Thus, a much shorter column will provide a satisfactory separation. Since the
separation is complete on the 3 m column, higher flow rates can be used to
reduce the analysis time.

SAQ 1
What are the main factors with which the resolution of peaks is related?

SAQ 2
State whether the following statements are true/false.

i) Column efficiency, is improved by increasing larger particle size packing.

ii) For a high efficiency a high molecular weight carrier gas should be used.

SAQ 3
What is the effect of increased column temperature on separation in general ?
103
Block 1 Solvent Extraction and Chromatographic Methods

3.3 INSTRUMENTATION
Upto this point we have refreshed some of the basic concepts of gas
chromatography and understood the significance of different terms affecting
the efficiency of separations. But the separations of the desired level are
achieved by the optimum performance of each component of the instrument.
Fig. 3.9 shows schematically, the essential components, of a chromatograph.

1. Carrier gas cylinder 6. Detector

2. Flow meter 7. Electronics

3. Sample injection chamber 8. Recorder

4. Column 9. Fraction collector

5. Oven

Fig. 3.9: Block diagram of a typical gas chromatograph.

Let us now discuss in brief, the construction and working of each component
of a gas chromatograph.

3.3.1 Mobile Phase (Carrier Gas)

The carrier gas is normally available in a compressed gas cylinder. It is used
to transport the vaporized solutes from the injection port through the column to
the detector. The carrier gas must be

i) inert,

ii) dry, and

iii) virtually free of impurities.

It must be inert to an extent so that no chemical reactions can take place with
any portion of the instrument, column or solute. It must be dry because water
can and usually does react with some portion of the column. This results in
loss of resolutions and tends to produce asymmetric or "tailing" peaks.
Unwanted compounds or ghost peaks may also appear. Another effect is a
net loss of sensitivity, that is, the ability to quantify small concentrations of
solutes. The dotted lines in Fig 3.10 show the effect of water in carrier gas on
104 column performance.
Unit 3 Gas Chromatography

Fig. 3.10: Effect of water on column efficiency.

Impurities such as hydrocarbons, oxygen as well as water can and usually will
contribute to unwanted noise levels and perhaps baseline drift. Additional
peaks are also seen in the chromatogram. Ideally, (ultra-pure gases
(99.9999%) should be used. Easily obtainable gases at 99.995% purity level
are usually satisfactory. Commercial grade gases can be used only if special
driers and scrubbers are used to reduce the impurities to a usable or
satisfactory level. Water and trace hydrocarbons can be easily removed by
installing a 5Aº molecular sieve filter between the gas cylinder and the
instrument. After two gas cylinders have been used, the sieve should be
regenerated by heating to 300°C for three hours with a slow flow of nitrogen.
Oxygen is more difficult to remove and requires a special filter, such as BTS
catalyst from BASF. The gases that are commonly used in GC as carrier
gases include hydrogen, helium, nitrogen and argon. Argon and nitrogen are
inert and are easily obtainable at the purity levels required and therefore are
the most popular carriers. Some detectors require specific gases and purity
levels for optimum performance. For example

Detector Carrier gas

Thermal conductivity Helium

Flame Ionization Helium or nitrogen

Electron capture Very dry nitrogen or argon, 5% methane

The solutes of interest in the sample may also dictate the choice of a
particular carrier gas. For example, in the analysis of hydrogen in air, nitrogen
would be the best choice of carrier. Moreover, there is a need of flow
controller.

 Flow controller and pressure regulator

The measurement and control of carrier gas, flow is essential for both
column efficiency and for qualitative analysis. The more constant the
flow rates, the more precise and accurate the results. The column flow
rates of carrier gas in conjunction with temperature will determine the 105
Block 1 Solvent Extraction and Chromatographic Methods

retention time of qualitative aspect of each sample solute. The flow rate
must have certain characteristics. It must be stable, reproducible and
convenient. Stability is necessary for accurate results, reproduceability
for precision and convenient parameter adjustment to optimise results.
For these reasons, a specially designed flow controller or pressure
controller is used to maintain precise and accurate flow rates for column
carrier gas or for auxillary gases needed for detector operation.

Fig 3.11 demonstrates as to what will happen if column carrier gas flow rate is
reduced from previous setting. All peaks have shifted to longer retention times
and have an apparent loss of peak height The base of each peak is also
wider, however, individual peak areas remain constant

Fig. 3.11: Effect of decreased flow rate.

The dotted lines in Fig 3.12 show the effect of the greatly increased flow rate.
The solvent does not change its position very much but the solutes are
squeezed towards the injection point. If in an analysis, two solutes are just
separated under normal conditions, the effect of the increased flow rate may
cause two solutes to elute together appearing as a single peak.

Fig. 3.12: Effect of increased flow rate.

The column flow rate is typically indicated by a rotometer. It is operated by the

volume of gas passing a ball in a tapered cell. The gas flowing past the ball
causes it to rise to a calibrated equilibrium position indicating the flow. Most
106 modern gas chromatographs incorporate rotometers as flow rate indicators.
Unit 3 Gas Chromatography

Fig 3.13 shows the design of a typical rotometer.

Fig. 3.13: Rotometer.

3.3.2 Columns
The column is the most important part of a gas chromatograph. It may be
called as the heart of the instrument where the separation takes place. Most
columns are usually constructed of glass or metal tubing filled with a carefully
sized granular solid support. The grains of the support are typically coated
with a relatively high boiling liquid phase. This liquid phase, often referred to
as “stationary phase” is the active portion of the column. It is the interaction of
sample solutes with the liquid phase that determines how much separation
can be achieved. Some columns do not use a liquid phase but rather employ
porous polymers or solid adsorbents to achieve separation. Each type of
column described has its own grouping of applications.

The type of column most commonly used is the packed column where the
tubing is filled with a coated solid support, polymer or absorbent. Glasswool
which is usually used to retain the packing within the column is placed on both
sides of column. The end of the column which is fitted to the sample
introduction system usually has 4 to 6 cm of glasswool, whereas the opposite
or the detector end has about 6 mm of glasswool. The schematic of a packed
column is shown in Fig. 3.14.

Fig. 3.14: Packed column. 107

Block 1 Solvent Extraction and Chromatographic Methods

In general, stainless steel columns are used, packed while straight to obtain a
uniform packing and coiled to facilitate long lengths. If coiled, the spiral
diameter should be at least ten times the column diameter to minimize
diffusion and racetrack effects. Packed column lengths vary from a few
centimeters to several metres. Common analytical columns are 1 to 3m in
length. Column diameters vary from 0.25 mm to 50 mm I.D. and larger. The
smaller the column diameter, the higher the column efficiency. Standard
analytical columns are of 3 mm to 6 mm I.D.

Another type of column is the open tubular column. This is usually 0.25 mm ID
tubing. A liquid phase is coated directly on the inside wall of the tubing;
therefore, no additional packing is needed. Due to small surface areas
involved and the resulting limited capacity for solutes, these columns are
necessarily long (20 to 50 m) and require the use of a specialized sample
inlet. Since capillary columns are not filled with particles, band broadening due
to eddy diffusion does not exist. The design of an open tubular column is
shown in Fig. 3.15.

Fig. 3.15: Open tubular column.

A comparison of packed and capillary columns given in Table 3.1 shows some
interesting differences. A major advantage of capillary columns is the total
plates obtainable. Plates per metre of column length are comparable with
packed columns but much longer columns are usable because capillary
columns have high permeabilities, i.e. they are “open tubes” with small
resistance.
Table 3.1: Comparison of Capillary and Packed Columns
Capillary Packed
1/16-in. 1/8-in 1/4 -in. 3/8-in. O.
O. D. O. D. O. D. D.
Inside diam., inches 0.01-0.02 0.047 0.065 0.155 0.315
Maximum plates/ft
1000 1000 800 500 300
Practical length Max.
total plates Amt. liquid 300 60 60 60 100
phase, % Liq. Film, 300,000 60,000 48,000 30,000 15,000
thickness, µ Mesh range
Permeability, 107 cm2. -- 3 5 10 20
Avg. linear vel, cm/sec/ 1 5 5 10 20
Avg. flow rate, ml/min
-- 100/20 80/100 60/80 20/40
Max. sample size, µl.
200-800 1.5 2 3.5 4.0
25 10 10 7 7
1-3 10 20 70 120

108 0.010 1.0 2.0 20 1000

Unit 3 Gas Chromatography

The process of separation can be considered as a series of partitions where the

vaporized solutes from the injection port go into solution with the liquid stationary
phase and are then revaporized. The affinity of the liquid phase for a sample
component determines the length of time for which each individual component will
remain in solution. After a number of partitions, various solutes will separate into
discrete bands and travel at different velocities through the column. The most
volatile solute usually passes through the column first and the least volatile or
highest boiling solute emerges last. Fig. 3.16 demonstrates these stages of
separation.

Fig. 3.16: Component separation within the column.

3.3.3 Stationary Phase Support and Liquid Phases

Chromosorb A: It is
After studying different types of columns, you would like to know about the actual used in preparative-
packing material inside them particularly their characteristics. This sub- section is scale gas
devoted to this particular aspect. The discussion will be taken up under the chromatography. It can
hold upto 25% liquid
following heads. phase. It has a good
mechanical strength
A. Stationary phase support. and its surface is not
highly adsorptive. It is
B. Liquid phases. available in mesh
ranges of 10/20, 20/30
C. Super selective liquid phases and 30/40. These allow
the use of long
D. Liquid phase percentage columns with a low
pressure drop.
E. Effect of column temperature

F. Porapak as packing material.

A. Stationary phase support Chromosorb G: It is

used for the separation
The purpose of the solid phase is to support a thin uniform film of liquid of polar compounds. Its
phase. An optimum support should have certain characteristics : low surface area,
hardness and good
 A large specific surface area - from 1 to 20 sq. metres per gram handling characterisitics
make it a good
 A pore structure with uniform pore diameter in the range of l0 µ or less replacement for highly
friable supports.
 Inertness - a minimum of chemical and adsorptive interaction with the
solutes
109
Block 1 Solvent Extraction and Chromatographic Methods

 Regularly shaped particles, uniform in size for efficient packing.

 Mechanical strength – it should not crush on handling.
Chromosorb P : It is
a calcined diatomic, No material has yet been described which fulfills all these requirements;
pink in colour and however, several suitable supports are commercially available. Usually one
relatively hard. Its must choose between inertness or efficiency (high surface area). The raw
surface is more
adsorptive than the material for most gas chromatographic support is diatomite also known as
other Chromosorb diatomaceous silica, diatomaceous earth, and kieselguhr. Diatomite is
grades and it has the composed of skeletons of diatoms, microscopic unicellular algae, which are
best efficacies for
hydrocarbons. primarily micro- amorphous hydrous silica, although some micro impurities
mainly metallic oxides are present. The individual dual diatom skeletons are
very small, highly porous and have a high surface area. There are five
forms or types of chromosorb which is Johns Manville’s registered trade
mark for GC support material i.e., chromosorb A,G,P,W and T. Each is
available either untreated or treated and in a variety of mesh ranges.
Supports are usually sized by screening through standard ASTM screens.
Mesh numbers refer to the number of openings per linear inch. Particles that
will pass through 60 mesh but not 80 mesh are referred to as 60-80 mesh.
The more uniform the packing, the smaller the Eddy difussion and more the
column efficiency. For 3 mm I.D. Columns,100/120 or 80/100 mesh support is
preferred, for 6 mm I.D. columns, 40/60 or 60/80 mesh is suitable.

The chemical composition of some of these supports is given in Table 3.2.

Table 3.2: Chemical Analysis of Supports
Chromosorb W: It is Firebrick Celite Chromosorb P Chromosorb
a flux-calcined
C22 545 W
diatomite support. It is
similar in performance SiO2 89.7 99.9 89.2 91.2
and properties to
Celite 545. It is white Al2O3 5.1 3.6 5.1 4.1
in colour and friable Fe2O3 1.55 1.65 1.56 1.15
compared to
chromosorb G. Its TiO2 0.30 0.30 0.30 0.25
surface is relatively
nonadsorptive and is CaO 1.30 1.75 0.90 0.40
used for the MgO 0.90 0.70 1.00 0.65
separation of polar
compounds.
B. Liquid phases
There is no fool-proof method for selecting the best liquid phase for a
particular separation. The right selection is based mainly on experience and /
or trial and error. Following are the basic requirements of a liquid phase.
 It should be a good absolute solvent for sample solutes. If solutes are
sparingly soluble in the liquid phase, they will elute rapidly and separation
will be poor.
 It should exhibit differences in solubilities for different solutes in the sample.
 It should have low vapour pressure (0.01 to 0.1 mm) at operating
temperatures for a reasonable column life.
 It should be thermally stable.
 It should be chemically inert toward solutes of interest at the column
110 temperature.
Unit 3 Gas Chromatography

The choice of liquid phase depends upon several considerations. The liquid
phase chosen depends on the composition of the sample. It is expected that
the type of solutes likely to be present in the sample is known before the
analysis is started. The more one knows about a sample, easier it is to
select the proper column and operating temperatures. For an efficient normal
separation, the liquid phase should be similar in chemical structure to the
solutes of the sample. For example, hydrocarbon compounds are best
separated with a hydrocarbon solvent. Paraffins are best separated on
squalane (a long chain hydrocarbon). Polar solutes like alcohols are best
separated on a polar - Hailcomid (an amide) solvent. Table 3.3 gives the
names of common liquid phases in GC separations.

Table 3.3: Recommended Liquid Phases by Sample Types

Classification of Compounds Stationary Phase

Hydroxy 2,4-Xylenyl Phosphate
Olefinas AgNO3/Benzyl cyanide
C1-C5 Dimethylsulfolane
Propylene Carbonate
C5-up Carbowax 20M
Polynuclear SE-30 on DMCS treated support
FFAP on DMCS treated support
PMPE (5 ring)
Ketones Lexan
FFAP
Nitrogen Compounds
Amines Dowfax 9N9/KOH
Amides Versamid 900
Ammonia Ethofat or Carbowax 600 cc
Chromosorb T
Nitriles Tetracyanoethvlated Pentaerythritol
FFAP
XF-1150
Organo-metallic FFAP
SE-30
Alcohols
C1-C5 Halleomid M-18 OL
Carbowax 600 or 1540
C1-C18 FFAP
Carbowax 20M
Di-Poly FFAP
QF-1
111
Block 1 Solvent Extraction and Chromatographic Methods

Aldehydes
C1-C5 Ethofat
C5-C18 Carbowax 20M

Alkaloids QF-1
SE-30
Amino Acid Derivatives
N-Butyltrifluoroacetyl esters DEGS/EGSS-X
Classification of Compounds Stationary Phase
Boranes Apeizon L
Essential Oils FFAP
(General) Carbowax 20 M
Esters Dinonyl phthalate
(Mixed)
Porapak Q
Ethers Carbwax 20 M
Glycols Porapak Q
Halogen Compounds Carbowax 20 M
QF-1 (FS-1265)
FFAP
Freons Dibutyl Tetrachlorophthalate
UCON Polar 2000
Hydrocarbons
Allphatic
C1-C5 Propylene Carbonate
Carbowax 100
Tributyl phosphate
C5-C10 Didecylphthalate
SE-30
Aromatics Tetracyanoethylated
Pentaerythrtiol
Dibutyl tetrachlorophthalate
Acids
C1-C18(free) FFAP
Bile and Urinary SE-30
Fatty Acid Methyl Esters DEGS
FFAP
Apleron L
TCEPE
EGSS-X
112
Unit 3 Gas Chromatography

Table 3.3: Recommended Liquid Phases by Sample Type (Cont.)

Classification of Compounds Stationary Phase

Water Porapak Q

Gases

Ar and O2 6-feet Activated Molecular

Sleve SA at -72°C

H2, O2, He, N2, CO, CH4 20-feet Molecular Sleve 13X

CO2, H2S, CS2, COS Silica Gel or Porapax Q

N2O, CO2, NO Porapak Q

Pesticides Dow 11

QF-1 (Fs-1265)

SE-30

OV-1 OV-17

Phosphorus SE-30

STAP

Silanes SF-96

FFAP

Steroids Stap

XE-60

QF-1 (FS-1265)

SE-30

OV-1

OV-17

Sugar Derivatives QF-1

Trimethylsilylethers SE-52

Sulphur Compounds Carbowax 20M

FFAP

Dinonylphthalate

Porapak Q

113
Block 1 Solvent Extraction and Chromatographic Methods

C. Super selective liquid phases

Certain liquid phases may be said to be super selective. Some of these

liquids depend on the formation of loose chemical adducts with particular
solutes to effect separation. Silver ions form loose adducts with olefins of
the type shown below

C
Ag +
C

Saturated solutions of silver nitrate in ethylene glycol, glycerol and

triethylene glycol or in benzyl cyanide have been used as selective
solvent for olefins. Benzyl cyanide has an advantage over the glycols
because it is non-hygroscopic and does not require moisture free carrier
gas. Parafins are not retained and pass rapidly through the column while
alkynes react to form siliver acetylides and remain on the column.
Temperature should be below 40°C, above this limit the adducts with
AgNO3 are not formed and solutions are unstable. Mercuric perchlorate
has been used to remove unsaturated by π- complexes with olefins,
alkynes and aromatics. The tetracyanoethylated pentaerythritol (TCEPE)
is highly selective for aromatics.

Liquid crystals are also used as selective phases. Liquid crystals are
intermediate between the crystalline solids and normal istotropic liquids .
They can exist in liquid crystal phase-smectic and nematic. The solid
passes from smectic liquid phase to the nematic and finally to the '
normal' liquid phase as temperature is raised. A typical formula for a
liquid crystal is:

RO N NO OR

whe re, R is CH2 or n  C6H13 or n  C7H15

The liquid crystals show a selective affinity for linear molecules. They will
retain p-disubstituted benzenes relative to o-and m-isomers.

D. Liquid phase percentage

The amount of liquid phase used should be just enough to coat particles
with a thin uniform layer. Too much liquid phase collects in pools
between particles and the efficiency of the column decreases. The
efficiency decreases drastically for liquid loading exceeding 30% on
diatomaceous earth supports. The retention time is proportional to the
grams of liquid phase present, and thus, low liquid loading means fast
analysis.

The change of retention time with % liquid loading is illustrated in Fig.

114 3.17.
Unit 3 Gas Chromatography

Fig. 3.17: Retention time vs % liquid phase.

Too low a liquid load can leave adsorbing sites on the support exposed.
This could cause irreversible adsorption or decomposition of the sample.
To avoid this very inert supports should be used with low liquid loadings.
Volatility of solutes should also be considered while choosing the amount
of liquid phase. Solutes of low volatality are best run on low loaded
columns ( 3 % or less) for materials such as steroids. Very volatile
solutes such as light hydrocarbons require high liquid loads ( 20% to
30%) since their solubilities in the liquid phase are low. The more the
liquid phase, the longer the time which is spent in the liquid, the better is
the partitioning.

E. Column temperature

The distribution constant (Kc) is very temperature dependent. In most

cases, an increase of 30°C will halve the Kc and thus, double the rate
of solute migration. The most immediate effect of temperature increase
is a decrease in analysis time as shown in Fig. 3.18. 115
Block 1 Solvent Extraction and Chromatographic Methods

Fig. 3.18: Retention time vs temperature

Generally, resolutions can be improved by lowering the temperature. In

the usual case, temperature is chosen by compromise not so high as to
impair resolution and not so low as to cause long retention times. In
selecting column temperature, one must also keep in mind the maximum
and minimum operating temperatures of the liquid phase being used.
Operating temperature should be above the melting point of the liquid
phase in order for it to function effectively.

F. Porapak

Porapak is a porous polymer composed of ethylvinylbenzene cross-

linked with divinylbenzene to form a uniform structure of a discrete pore
size. Porapak column contains only the porous polymer beads screened
to a definite mesh range and no liquid phase or solid support is used.
The porous polymer beads serve the function of both the liquid phase
and the solid support. Apparently, the solute molecule positions directly
from the gas phase into the amorphous polymer. There are six Porapak
materials available like type P,Q,R,S,T, and N. Type P and Q are non-
polar phases type, and R,S,T and N are moderately beads. Porapak N
has been developed for separation of formaldehyde mixtures and for the
separation of acetophenone and ethylene. All beads are available in
various particle sizes ranging from 50/80 mesh to 150/200 mesh. All the
polymers are stable to 250°C except Porapak T which is stable upto
200ºC.

Porapak is used primarily for efficient separations of a wide variety of relatively

low molecular weight solutes. Porapak columns should be conditioned prior to
use to remove residual solvents from the polymerization process. The utility of
the material is very well demonstrated by a variety of a separations shown in
116 Fig. 3.19. The conditions are mentioned.
Unit 3 Gas Chromatography

Fig. 3.19: Applications of porapak column.

3.3.4 Detectors
Once the sample is injected, separated and separated solutes are emerging
from the column, a method of detection is required. The selection of a detector
will depend upon sample’s molecular structure and concentration ranges for
the components of interest. The proper choice of detector along with the
column can optimize the results for the investigator.

Since chromatographic detectors differ greatly in the principle under which

they operate, it is difficult to compare them. Certain characteristics, however,
are indicative of the usefulness of a detector. Some of these are mentioned
below.

 Adequate selectivity

 Sensitivity and detectability 117

Block 1 Solvent Extraction and Chromatographic Methods

 A linear response to solutes that extends over several orders of

magnitudes

 Operative in a temperature range from room temperature to at least

400°C

 High reliability and ease of use

 A short response time that is independent of flow rate

 Similarity in response towards all solutes

 Non-destructive for solutes

 Noise and ninimum detectable quality

Needless to say, no detector exhibits all of these characteristics and it seems

unlikely that such a detector will ever be designed. Detectors may be
classified as "integrating" or “differentiating” type. An integrating detector gives
a response proportional to the total mass of solute in the eluted zone.

As shown in Fig. 3.20, when pure carrier gas passes through the detector, the
strip charge recorder shows a straight line. As a solute zone passes through
the recorder, pen moves across the chart by a distance proportional to the
total mass of the component in the zone. When another solute zone is eluted,
the pen moves further across the chart. The chromatogram produced by
integrating detector consists of series of steps in which the distance between
consecutive level portions of the curve is proportional to the total mass of the
solute corresponding to that step. The titrating burette is an example of an
integrating detector.

Fig. 3.20: Integral chromatogram.

A differentiating detector gives a response proportional to the concentration or

mass flow rate of the eluted component. The most familiar example of a
detector responding to concentration is the thermal conductivity detector
(TCD). The most familiar example of a detector responding to mass flow rate
is flame ionization detector (FED). The chromatogram produced by a
differentiating detector consists of a series of peaks, each of which
corresponds to a different solute. The area under each peak is proportional to
the total mass of that solute (see Fig. 3.21). Differentiating detectors are most
118 commonly used because of their convenience and accuracy.
Unit 3 Gas Chromatography

Fig. 3.21: Differential chromatogram.

Let us first consider a detector responding to concentration. Ideally, we have

the following relationship

R = K1C … (3.13)

where, R is detector response in millivolts,

K1 is a constant of proportionality, and

C is the concentration of the solute passing through the detector.

If the detector response is plotted against time t, we get a curve shown in Fig.
3.22.

Fig. 3.22: Detector response vs. time.

If the shaded area under curve is A, substituting the value of R as given in Eq.
3.13, we get
t2 t2

A  K Cdt  K  Cdt
t1
1 1
t1
(3.14)

Now let us consider a solute zone in the form of a plug in which the component
concentration is constant (Fig. 3. 23) and equal to M/V where, M is the total
mass of a solute in the plug and V is the plug volume.

Fig. 3.23: “Plug” flow. 119

Block 1 Solvent Extraction and Chromatographic Methods

If C is constant, then
t2
A  K 1C dt  K 2 C t2  t1   K1 Ct
 … (3.15)
t1

But C = M/V, therefore,

A = (K1 M/V) ∆t … (3.16)

However, V= F, t, where F is carrier gas flow rate. Thus, ∆t e= V/F

And A = (K1 M/V)(V/F) = K1M/F … (3.17)

Thus, finally we obtain

A = K1M/F

Eq. 3.17 shows that peak area is directly proportional to the total mass of
component. The above equation also shows that for a detector responding to
concentration, the peak area is inversely proportional to the carrier gas flow
rate. Thus, for accurate quantitative analysis with TCD, the flow rate must be
kept constant. For detectors responding to mass flow rate (dm/dt), such as the
FID, we have

 dm 
R  K2   … (3.18)
 dt 

where, K2 is a constant of proportionality, m is the instantaneous mass of

component within the detector, and R and t have their earlier meaning. By
similar reason:
t2 t2 t2
A t Rdt  t K 2 dm/dt  dt  K 2 t dm/dt  dt … (3.19)
1 1 1

Cancelling dt within the integral sign, and integrating, we obtain

A=K2m … (3.20)

Eq. 3.20 shows that for a detector responding to mass flow rate, the peak
area is proportional to the total mass of the eluted solute. However, unlike the
concentration detector, the peak area for a mass flow detector is independent
of carrier gas flow rate. Constant flow rate is not as critical for FID as for a
TCD.

 Detector Sensitivity

The sensitivity of a detector depends upon its operating principle.

Sensitivity or detectability has been defined as the ratio of the smallest
detectable quantity measured at twice the noise level to the peak width
at base in seconds. For example, a 0.1 ng (10–10 g) sample with a 60 sec
peak width at base would show a sensitivity of 1.6 × 10 –12g/sec If the
column temperature is lowered and the same sample now has a 180 sec
peak width at base (due to longer retention time) the resulting sensitivity
would be 0.55 × 10–12 g/sec.
120
Unit 3 Gas Chromatography

 Detector response

The response of a detector is the quantity of a signal generated by a

given amount of the solute.

 Noise and minimum detectable quantity

Detector sensitivity can be made as large as desired when the electrical

output of a detector is increased to almost any desired value by
electronic amplification. However, electrical noise inherent in the detector
and electronics is also amplified and a point is reached where the noise
level is high enough to obscure the detector’s response. Therefore, the
noise level limits the concentrations or mass flow rate of solutes that can
be detected.

Fig. 3.24: Minimum detectable quantity.

The minimum detectable quantity is that amount which gives a detector

response equal to twice the noise level.

Linear detector range

Accurate quantitative analysis depends upon a linear relation between

concentration and detector response. The linear range of a detector may be
defined as the ratio of the largest to the smallest concentration within which
the detector is linear. There is a variety of detectors used in a gas
chromatograph. Here, it may not be possible to go into the details of the
different detectors. The design, working and utility of some of the important
detectors may be essential to know to make a proper choice of the detector.
Keeping this in mind, some of the detectors that are being briefly discussed
here are

1. Thermal Conductivity Detector (TCD)

2. Flame Ionization Detector (FID)

3. Electron Capture Detector (ECD)

4. Helium Ionisation Detector (HID)

5. Flame Emission Detector (FED)

1. Thermal Conductivity Detector (Katharometer) TCD

The thermal conductivity detector is based on the principle that a hot

body will lose heat at a rate which depends upon the composition of the
surrounding gas. Thus, the rate of heat loss can be used as a measure
of the gas composition. The TCD operates by comparing the thermal 121
Block 1 Solvent Extraction and Chromatographic Methods

conductivity of a sample component as reference against the carrier gas.

Fig. 3.25 shows a typical TC cell consisting of a spiral metal filament
supported inside a cavity within a metal block.

Fig. 3.25: Schematic of TC cell.

The filament is made of a material whose electrical resistance varies

greatly with temperature. It has a high temperature coefficient of
resistance. A constant current is passed through the filament causing its
temperature to rise with pure carrier gas. With pure carrier gas flowing
the heat loss is constant and thus, the filament temperature is also
constant. If the gas composition changes, e.g. when solute peak
emerges, the filament temperature changes, causing a corresponding
change in electrical resistance. It is this resistance change which is
measured by a Wheatstone bridge circuit (Fig. 3.26).

Fig. 3.26: T C Wheatstone bridge circuit.

The TCD is universal i.e., it will respond to any solute that has a thermal
conductivity different than the carrier gas. It is mainly used for analyzing
inorganic gases such as the components of air and its impurities. The
sensitivity ranges from large percentages of components down to a few parts
per million.

2. Flame Ionisation Detector (FID)

In operation as the solute elutes from the column, it is mixed with hydrogen
and then combusted. In the combustion process, transitional ions are
122 created that can be collected by a metal conductor.
Unit 3 Gas Chromatography

Fig. 3.27: Principle and structure of flame ionisation detector.

These collected ions cause an electronic signal which can be amplified and
displayed on a recorder. Ideally, the final combustion products are carbon
dioxide and water so that they can be safely vented to the atmosphere. The
number of these transitional ions formed is proportional to the total number
of solute molecules passing through the detector. The FID will respond to
organic compounds only i.e., those solutes that have a carbon-hydrogen
bond and are combustible. Table 3.4 lists the solutes for which FID does not
respond. The lack of response to air and water makes the FID especially
suitable for the analysis of air pollutants or aqueous samples such as
alcoholic beverages, biological materials etc. Similarly, the absence of a
“solvent peak” makes carbon disulphide a convenient solvent for use with
the FID.

Table 3.4: Compounds Giving Little or No Response in the Flame

Ionization Detector

He CS2 NH3
Ar COS CO
Kr H2S CO2
Ne SO2 H2O
Xe NO SiCl4
O2 N2O SiHCl3
N2 NO2 SiF4

In addition to normal carrier gas , auxiliary gases of hydrogen and air are
needed to fuel the flame. The hydrogen flame, being perhaps the most
popular detector in gas chromatography, is extremely sensitive and under
proper conditions, the detector is capable for responding to a few parts per
billion of the solute component. In general, good sensitivity and stability are
obtained with a carrier gas flow of 30 cm3/min, hydrogen flow of 30cm 3/min
and air flow at 300 cm3/min. 123
Block 1 Solvent Extraction and Chromatographic Methods

Fig. 3.28: FID sensitivity vs H2 flow.

The FID has the widest linear range of any detectors in common use (106 to
107). The combination of high sensitivity and wide linear range makes the
FID the choice in trace analysis.

3. Electron Capture Detector (ECD)

The characteristic features of an ECD are as follows:

i) Specific

ii) Selective to halogens

iii) Sensitive to a few parts per trillion

The electron capture detector measures the loss of signal rather than a
positively produced electrical current. As the nitrogen carrier gas flows
through the detector, a tritium or Ni63 source (characteristics given in Table
3.5) ionizes the nitrogen molecules and slow electrons are formed. The slow
electrons migrate to the anode under the fixed voltage which is formed as “cell
voltage”. The slow electrons produce a steady current. If a sample
containing electron absorbing solutes is introduced, this current will be
reduced. The loss of current is a measure of the amount and electron affinity
of the solute.

Table 3.5: Isotope Characteristics

3 63
Characteristics H Ni
Half Life 12.5 125 years
Type of Decay β β
Particle Energy 18 kev 67 kev
Specific Activity 9,800 curie 5 curie/gram
Physical Properties Gas Solid
Fall Temp. Limitation 220° C 200° C
Radio Toxicity Very low Moderate
Cost Per Curie $0.024 $17.5
124
Unit 3 Gas Chromatography

The ECD is extremely sensitive to certain solutes, such as alkyl halides,

conjugated carbonyls, nitriles, nitrates and organometals, but is virtually
insensitive to hydrocarbons, alcohols, ketones etc. Selective sensitivity to
halides makes ECD especially valuable for the analysis of pesticides, Both
H3 (tritium)and Ni63have been employed in the ECD. The linearity
response for the analysis of lindane (a pesticide) using H3 and Ni63 is
shown in Fig. 3.29.
Peak Height Response

Fig. 3.29: H3 and Ni63 linearity response.

4. Helium Ionisation Detector (HID)

Some of the characteristics of this detector are given below.

i) Universal

ii) Photo-ionization

iii) Sensitive to fractional parts per million

The helium ionisation detector is a universal detector except of course, it will

not sense helium. The combination of the tritium beta radiation and high field
gradient (4000 volts per cm) raise the helium to a metastable state with an
ionisation potential of 19.8 eV. All solutes having a lower ionisation potential
will be ionized giving rise to a positive signal. The HID was developed for the
ultratrace analysis of permanent gases.

5. Flame Emission Detector (Alkali Flame Detector)

The flame emission detector consists of a normal flame detector plus the
addition of a small alkali salt pellet placed at the burner set with precise
control of the hydrogen and air flow rates. This detector can be made to give
an enhanced response to phosphorus containing compounds. The three major
differences between the standard FID and alkali flame detector are the alkali
salt tip, the air flow rate and the electrode configuration. Both CsBr and
Rb2SO4 have been used with good results. These salts give a selectivity of 125
Block 1 Solvent Extraction and Chromatographic Methods

5000 L for phosphorus and 50/L for nitrogen over normal hydrocarbons.
Alkali flame detectors operate at air flow rate of approximately 130 cm3/min
which is significantly below that of a normal FID (300-400 cm3/min). Since
constant air and hydrogen flow rates are important for the stability of this
detector, flow controllers are required. The alkali flame detector has been
used extensively in the analysis of organo-phosphorus pesticides and for
nitrogen containing solutes. The design of the detector is given in Fig. 3.30.

Fig. 3.30: Alkali flame detector.

SAQ 4
What are the essential requirements of a carrier gas? How are water and
hydrocarbon impurities removed from the carrier gas before its use?

SAQ 5
What are the major advantages of open tubular columns over packed
columns?

SAQ 6
What will be the effect on separation, in general, if there is an excess liquid
loading in the column?

SAQ 7
What is porapak? What is the advantage in its use in the column?

SAQ 8
What is the minimum detectable quantity of a detector? How is it affected with
the noise level of the detector?onfigurations of C–2 carbon atoms in (+)-
isoserine and ()-glyceric acid.

126
Unit 3 Gas Chromatography

SAQ 9
What type of detector will you suggest for the analysis of the following by gas
chromatography?
i) Organochlorine pesticides
ii) Inorganic gaseous pollutants in air
iii) Organophosphorus pesticides

3.4 SAMPLING AND INTRODUCTION OF SAMPLE

INTO GC UNIT
For analytical results to have any validity or meaning, adequate sampling
procedures should be adopted. Sampling is the process of extracting from
large quantity of material a small portion which is truly the representative of
the composition of the whole material under scrutiny. Proper sampling in GC
assumes a particular significance because here we deal with the analysis of
gaseous substances or components which are volatile.

The sampling process involves several steps:

1. Removal of an aliquot from a bulk quantity which is representative of that

bulk.

2. If necessary, alteration of the physical state of the sample (e.g. that of

gas) so that it can be manipulated.

3. Transportation to the laboratory for analysis.

4. Storage prior to analysis.

5. Introduction of the sample into the GC unit for analysis.

3.4.1 Hazards in Sampling

In the sampling, handling and storage of the gaseous and volatile materials,
there are some hazards involved and due precautions should be taken. The
analyst must, as a general rule, try to obtain a prior knowledge of the material
which is expected to be analysed. The analyst must wear adequate protective
clothing during sampling. Adequate antidodes etc must be kept ready before
proceeding for sampling work. No naked flame should be allowed near a
sampling area, if the sample is known to contain inflammable components.
Many gases/ vapors may be toxic. Pressure changes may release excessive
quantities of gas. Spontaneous ignition of inflammable gases may occur.
Sudden vaporisation of liquefied gases may take place. Volatile and
inflammable liquids may pose sufficient dangers to a careless analyst. Using a
facemask/ goggles during sampling is highly recommended.

3.4.2 Introduction of the Sample into the GC Unit

The introduction of the sample into the GC unit is the first step in the
chromatographic process. Its efficiency, therefore, gets reflected in the overall 127
Block 1 Solvent Extraction and Chromatographic Methods

efficiency of the separation procedure and the accuracy and precision of the
qualitative and quantitative results. In this regard, some aspects given below
need to be considered.

 Sample size.

 Injection systems.

 Sample injection port.

A. Sample size

The sample volume may vary depending upon the amount and rate of
evaporation of the sample in the sample port in the case of liquids/
solutions of solids or on the pressure of gas/ vapor sample contained in
the sample container. In the case of gases, it is a common practice to
vary the amount of sample by introducing different volumes at a constant
pressure. The sample size will obviously be related to the kind of solute-
stationary phase interaction (linear or non-linear) and consequently to
the shape of the final chromatographic peak obtained.

B. Injection systems

Various kinds of injection systems have been incorporated in commercial

GC units. Almost as important as column choice is the choice of injection
method. The various approaches are mentioned below:

i) Direct injection: Injection into the packed column.

ii) Split injection and splitless injection: These are adopted for
capillary GC.

iii) Solid injection (moving needle): In this case, the solvent is

evaporated before injection, resulting in improved separation of
fast eluting components.

iv) On-column injection: The sample is introduced jnto the column as a

liquid, reducing sample decomposition and giving reliable
quantitative analytical results. This is considered as being an ideal
solution for quantitative capillary GC.

v) Valve injection: Reproducibility of the valve injection volume is

superior to any other injection technique. Valve injection can easily
be automated which makes it very appropriate for routine on - line
analysis.

vi) Thermal desorption cold trap injection: Gaseous samples are

concentrated on an adsorbent tube. The components are
concentrated on a cold trap and then injected into the GC unit by
rapid heating.

vii) vii) Purge and trap injection: Aqueous samples are purged with
inert gas and the components are concentrated on a cold trap.
Entrained water is frozen out in the condenser. The components
are injected by rapid injection of the cold trap for multidimensional
128
Unit 3 Gas Chromatography

analysis, multiple switching intelligent controllers are available. The

sample is pre-separated on a pre-column. Coeluting components
are trapped on a cold trap and reinjected into a column for further
separation and analysis.

C. Sample injection port

The general requirements for an injection port are controlled

temperature, low volume and inertness in construction. The temperature
must be maintained within the proper range to vaporise all the solutes
instantly but not too hot to cause thermal decomposition of labile solutes.

The entire injection port must also be swept with carrier gas to ensure
that all of the vaporized solutes are moved quickly into the column. This
means that its internal portion must be kept to a minimum volume. The
materials of construction of the injection port should be such that no
reaction with the solutes occurs. It must be as inert as much as possible
to minimize any possibility of catalytic decomposition because of contact
with exposed surface area.

The syringes used to inject samples are usually of small volumes. The
normal ranges are from zero to 1 microlitre, zero to 5 microlitres and
zero to 10 microlitres. In the one microlitre syringe, the sample is totally
confined to the

needle and can never be observed by the operator. For the other two
syringes, the sample enters and its level can be directly measured in the
barrel of the syringe. The barrel is calibrated in microlitres and fractions
of microlitres.

Fig. 3.31: Sampling syringe.

Sample introduction for liquids on packed columns is often accomplished

with a microsyringe through a self-sealing silicon septum as shown in
Fig.3.32.

Syringe
needle

Fig. 3.32: Simplified injection port for on-column injection. 129

Block 1 Solvent Extraction and Chromatographic Methods

In this figure, the column is lined up colinearly with the syringe needle
providing for two possible injection modes: on-column or flash
vaporization.

For on-column operation, the column is positioned as shown in Fig. 3.32

with the column packing beginning at a position just reached by the
needle. When the syringe is pushed as far as it will go into the port, its
contact will be delivered into the first part of the column packing, ideally
on a small glass wool plug used to hold the packing in the column.

In the second configuration, the column is placed so that its front end
and its packing barely extends into the injection port and cannot be
reached by the syringe needle. Efficient sampling for this configuration
requires that the solutes evaporate quickly (flash vaporization) when
injected into the port.

For samples that exist as gases under normal conditions, a gas

sampling valve, shown in Fig 3.33, is used for sample injection.

Fig. 3.33: Gas sampling valve.

As carrier gas passes through the valve on the way to the column, a
sample can be filling the sample loop. When the actuator is turned the
carrier gas stream is redirected through the sample loop and sweeps the
sample into the column. The loop itself may have a volume as low as one
quarter of milliliter or as large as 10 milliliters. The choice of loop size is
made to be compatible with the analyst's experimental needs.

3.5 APPLICATIONS
Gas chromatography has numerous applications. It is used for both qualitative
and quantitative analysis. Before discussing applications, it should be clear in
130 mind that the column in a gas chromatograph is a separating tool. But the gas
Unit 3 Gas Chromatography

chromatograph as a whole with a detector and a recorder gives the information

about the eluting bands. Thus, the instrument provides both qualitative and
quantitative information about the components of a sample. The discussion
under this section will be taken up under three heads

1. Identification of compounds (Qualitative Analysis)

2. Quantitative analysis
3. Some examples of applications

3.5.1 Identification of Compounds (Qualitative Analysis)

It should be known that with a particular column if all the variables such as
temperature and flow rate are carefully controlled, the retention time or
retention volume of a solute is a property of that solute, just as its boiling point
or refractive index is a property. This amounts to the fact that retention
behaviour can be used to identify a compound. If one wishes to identify the
unknown compounds in a mixture, the standards are run under identical
conditions and by matching the chromatograms of unknowns and standards,
the former are identified. At this point it may be important to point out that such
type of identification is not the strong point of gas chromatograph. There are
instruments like mass spectrometer which can provide more information about
the unknown. Basically GC is a good separation technique. If it is hyphenated
with instruments like mass spectrometer or infrared spectrometer, the system
provides an ideal set up for identification of the unknowns present in a mixture.
The mass and the ir spectra provide valuable information about the column
eluants. There are various other approaches which are used in conjunction
with GC to identify the separated components. The discussion on these is not
being taken up here.

3.5.2 Quantitative Analysis

The chromatogram obtained on a recorder chart can be used to measure
quantitatively the concentration of components in a mixture. In general, three
methods are used for quantitative evaluation:

i) Area normalization method: In this method, it is assumed that the

entire sample is eluted from the column. The area of each peak is
measured and percent composition is obtained by dividing the individual
peak area by the total area of all the peaks and multiplying by 100. The
value so obtained will be acceptable only if the detector response is the
same (particularly for FED) for all the components of the mixture. If not,
the detector response factor for each component needs to be
established and appropriate corrections made in the measured areas.

ii) Internal standardization method: In this method, known amounts of

sample and standard are mixed and chromatographed. The peak areas
for sample component and for standard are measured and ratios of both
peak areas are determined. Either area ratios are plotted against weight
ratios to obtain a graph or area ratios for unknown are compared directly
with those for the known amounts. Thus, accurately known quantity of
the internal standard is added to the unknown sample and this mixture is
chromatographed and area ratios are measured. 131
Block 1 Solvent Extraction and Chromatographic Methods

The requirements for a suitable internal standard are summarized as follows:

 The standard must be a compound, which is well separated from all

components of the mixture being analyzed.

 The standard must not react with any component of the sample, nor it
should influence the physical properties of the other components, e.g.
volatility.

 It should yield a symmetrical chromatographic peak.

 It must be chemically similar to the unknown.

 It should have concentration comparable to the components of

interest.

iii) Comparison method: In this method, a synthetic mixture containing

known quantities of the components of interest in the range of
concentration expected in the unknown sample is prepared and
analyzed. The values for the peak areas for different known volumes of
synthetic blends are estimated and a calibration curve is plotted An
exact quantity of the unknown sample is then injected and the peak
areas so calculated are used to read from the calibration curve the
concentration of the component in the unknown mixture.

3.5.3 Some Examples of Applications

There are too many applications of GC to be listed here. It has been estimated
that perhaps 20 % of the known chemical compounds and almost 80 % of the
organics can be analysed by GC. Some of the important applications from
different areas are being discussed to illustrate the versatility of GC.
1. Bacterial identifications
Long chain fatty acids found in the bacterial cell can be used to
distinguish between various microorganisms. Fatty acids with chain
length from Cl0 to C20 can be separated and estimated on a 3 m glass
column of 2 mm internal diameter packed with 3% SP–2100 DOH at
oven temperature of 150°C to 225°C with nitrogen gas at a flow rate of
20 rnL/min.
2. Environmental analysis

a) Water analysis: The organic pollutants in water are concentrated

from water samples by solvent extraction or by purge and trap
technique. The volatile pollutants are analysed on 80/100-mesh
carbopack C/0.2 % carbowax 1500 column.

b) Air analysis: The analysis of organic vapours in the industrial air

environment for the assessment of exposure to workers is done as
follows:
i) Organic vapours are collected on a charcoal adsorbent with a
portable pump.
ii) Desorption from charcoal is done in a closed vial with carbon
132 disulphide.
Unit 3 Gas Chromatography

iii) Analysis of the desorbed sample is done on a GC using a 6

m, 3 mm internal diameter S.S column packed with 10 % SP–
1000 by temperature programming from 100°C to 200°C. By
this method, pollutants such as vinyl chloride, xylenes and
aromatic hydrocarbons can be estimated.

c) Clinical and toxicological analysis: Toxicologists have

recognised the usefulness of GC for the analysis of toxic
substances. The analysis of lidocaine and diphenhydramine has
been done using flame ionisation detection. A 15m x 0.25mm i.d.
5% methylsilicone (DB-5) column has been used, temperature
programmed from 180°C to 230°C at 5°C min–1 and helium used
as a carrier gas.

d) Forensic toxicology: It is highly specialised branch of analytical

chemistry concerned primarily with the analysis of specimen from
different organs of the human body for toxic substances. A simple
GC system utilising four columns and three liquid phases ( SE–30 ,
Hallcomid M-18, and Carbowax 6000), complemented by a direct
solvent extraction scheme designed to detect common poisons,
drugs, and human metabolites to a sensitivity limit of 2 µg/ml in
blood, urine and tissue is developed.

SAQ 10
For the identification of components present in a mixture, it is recommended
to use GC-MS instead of GC only. Explain why.

SAQ 11
Name the three methods which are used for quantitative analysis by gas
chromatography.

3.6 SUMMARY
Gas chromatography where gas as the mobile phase carries away the sample
through a column, separations result and the different components appear at
different time. Depending upon the nature of the stationary phase, GC can be
classified into two categories as GSC and GLC. The present unit basically
discusses the topic in three parts.

The first part is devoted to a recapitulation of fundamentals of gas

chromatography. The significance of different terms and concepts has been
mainly dealt under two heads-column efficiency and solvent efficiency.

The second part deals with instrumentation including the characteristics of

carrier gas. It begins with carrier gas followed by discussion on the different
types of columns and the stationary phases contained in these columns and
ends with a critical evaluation of commonly used detectors with a gas
chromatograph. 133
Block 1 Solvent Extraction and Chromatographic Methods

The last part deals with utility of the technique for analysis. Here, the
importance and methodology of sampling and the introduction of sample in the
GC unit are described. Some aspects of use of the technique both for
qualitative and quantitative analysis are discussed. It is difficult to describe
even some of the representative applications of gas chromatography because
of their numbers. However, some typical applications from toxicology and
environmental analysis are cited.

3.7 TERMINAL QUESTIONS

1. Name the parameters which have to be given due consideration to
improve the column efficiency.

2. Comment on the effect of temperature on solvent efficiency.

3. What should be the characteristics of an optimum support for the liquid

phase?

4. What are the basic requirements of a liquid to be used as a stationary

phase in a gas chromatograph?

5. Mention the characteristics of a detector which are indicative of its

usefulness.

6. What are the general requirements of an injection port in a GC unit?

7. How are the aromatic hydrocarbons determined in air of an industrial

area using gas chromatography? Mention the different steps.

3.8 ANSWERS
Self Assessment Questions
1. The resolution of chromatographic peaks is related to the following two
factors:

i) Column efficiency, and

ii) Solvent efficiency.

2. i) False

ii) True

iii) False

3. Increased column temperature leads to faster separation but poor

resolution.

4. The carrier gas should be inert, dry and virtually free of impurities. It
should not react with any portion of the instrument and solutes to be
separated. Water and hydrocarbon impurities are removed by installing a
molecular sieve filter between the gas cylinder and the instrument.

5. A major advantage of open tubular columns over packed columns is the

total number of plates that can be obtained. In open tubular columns,
134
Unit 3 Gas Chromatography

much longer columns can be used because they have high

permeabilities that is they are open with small resistance.

6. The amount of liquid phase used should be just enough to coat particles
with a thin uniform layer. Excess of liquid phase collects in pools
between particles and the efficiency of the column decreases.

7. Porapak is a porous polymer composed of ethylvinylbenzene cross-

linked with divinylbenzene to form a structure of a discrete pore size.
Porapak column contains only the porous polymer beads of a definite
mesh range and no liquid phase or solid support is used. The porous
polymer beads serve the function of both liquid phase and solid support.

8. The minimum detectable quantity of a detector is that amount which

gives a detector response equal to twice the noise level.

9. The detectors suggested are

i) For organochlorine pesticides–Electron Capture Detector(ECD)

ii) For inorganic gaseous pollutants in air–Thermal Conductivity

Detector (TCD)

iii) For organophosphorus pesticides–Alkali Flame Detector.

10. Gas chromatography is a very versatile technique as regards the

separation of the components of a mixture. For identification, one of the
best criteria is retention time/ volume. It is not entirely satisfactory. Mass
spectrometer is a very good technique for actual identification.
Therefore, if the two techniques are properly hyphenated the
experimental set up becomes a very powerful tool for the identification of
unknown compounds present in a mixture.

11. The three methods which are used for the quantitative analysis by gas
chromatography are

i) Area normalization method,

ii) Internal standard method, and
iii) Comparison method.

Terminal Questions
1. The different parameters which require due consideration to improve the
column efficiency are given below.

i) Particle diameter

ii) Flow rate

iii) Nature of carrier gas

iv) Type of liquid phase

v) Amount of liquid phase

vi) Pressure 135

Block 1 Solvent Extraction and Chromatographic Methods

vii) Temperature

viii) Column diameter

2. Solvent efficiency is measured by α, the relative retention. It is ratio of

adjusted retention time or partition coefficients.

X 2 K
α =  2
X 1 K1

α is temperature dependent, it may be constant over a limited

temperature range. The distribution constant decreases with increasing
temperature i.e, the fraction of the solute in the gas phase will increase
and hence, the elution time will decrease. This results in decreased
separation since it is the liquid phase which performs the separation. No
separation occurs in the gas phase. To achieve better separations, lower
temperature should be used. Lower temperatures means more liquid
phase interactions, more separation and longer time of analysis.

3. The characteristics the optimum support for the liquid phase are as
follows:

i) A large specific surface area (1 – 20 sq m/ g).

ii) A pore structure with uniform pore diameter in the range of 10µ or
less.

iii) Inertness that means a minimum of chemical and adsorptive

interaction with the solutes.

iv) Regularly shaped particles, uniform in size for efficient packing.

v) Mechanical strength–it should not crush on handling.

4. The basic requirements of a liquid phase are as under

i) It should exhibit different solubility for the components present in

the mixture.

ii) It should have low vapour pressure (0.01–0.1mm) at operating

temperatures for a reasonable column life.

iii) It should be thermally stable.

iv) It should be chemically inert towards solutes of interest at the

column temperature.

5. The following characteristics are indicative of the usefulness of a

detector.

i) Adequate selectivity

ii) Sensitivity and detectability

iii) A linear response to solutes over several orders of magnitude

iv) Operative in a temperature range from room temperature to at least

136 400ºC
Unit 3 Gas Chromatography

v) High reliability and ease of operation

vi) Short response time which is independent of flow rate

vii) Similarity in response towards all solutes

viii) Nondestructive for solutes

ix) Noise and minimum detectable quality.

6. The general requirements of an injection port are that it should be able to

maintain it at constant temperature. It should have low volume capacity
and made of material which is quite inert to different substances.

7. The analysis of aromatic hydrocarbons in the industrial environment is

done by following under mentioned steps.

i) Organic vapours are collected on a charcoal adsorbent using a

portable pump.

ii) Desorption from charcoal is done in a closed vial with carbon

disulfide.

iii) The analysis is done using a gas chromatograph.

Further Reading
1. Quantitative Analysis, By R. A. Day Jr. and A. L. Underwood, Prentice
and Hall.

2. Principles of Instrumental Analysis, By D. A. Skoog, F. J. Holler and T. A.

Nieman, Thomson.

3. Instrumental Methods of Chemical Analysis, By G. W. Ewing, McGraw

Book Company.

4. Basic Concepts of Analytical Chemistry, By S. M. Khopkar, Wiley

Eastern Limited.

5. Analytical Chemistry, By G. D. Christian, John Wiley and Sons.

6. Instrumental Methods of Analysis, By H. H. Willard, L. L. Merritt (Jr) , J.

A. Dean and F. A35

7. Settle (Jr).

137