.

••
••
••
70 ••
Temperature Programming ••
Isothermal separation. ••
If the oven temperature is kept constant during the separation, the process is •
called isothermal GC chromatography.
One should know that the GC separation occurs inside the column where
the temperature is strictly controlled by the temperature of the oven. Oven
.••
temperature means column temperature and vise versa, the terms are used
interchangeably. The temperature ofthe column influences very strongly ••
the efficiency of the separation.
••
Column temperature is determined on the basis ofthe boiling points ofthe
samples' components. If the oven temperature is higher than the boiling

points ofthe eluants, they reside mostly in gaseous- mobile phase, elute too

.•
fast and separation is not efficient. If the temperature is too low, compounds
reside mostly in the stationary phase, elute too slow, peaks are broad and
••
separation is not efficient. Usually the oven temperature is below or close to
the lowest boiling point of the sample components.
••
When the range of the boiling points of the mixture exceeds 100 0 C, it is
••
very difficult to choose a proper temperature for the column. In such a case
new method is introduced in which the column temperature varies during
••
the process of separation.

••
Variable oven temperature
•
••
70
., ~
...,.,

~
,..

III
~
-

.,
~

lit 71

.,.,...

III Temperature programming is the controlled change ofthe column

temperature during the process of analysis. This method is extremely
effective in the separation of complex mixtures with wide range of boiling

...... points .

If during the process of separation the temperature of the column

....,­ fluctuates in a controlled manner the process is called temperature

programmed gas chromatography ( T P CG) . T P GC is some times

....,- acknowledged as a separate analytical technique, however it is a natural

development of gas chromatography where the separation occurs at non­

.,.,...
isothermal conditions .
Although the application of T P G C is relatively simple, the theory
of this techniques leads to complicated equations and difficult analysis. As

.,., it is in other chromatographic techniques, the theory of the process is not

yet fully developed, and the equations in use now are empirical, derived

.,., from experimental data .

One can note that the temperature increases over the entire length of
the column, and thus special engineering care is taken to insure that the

.,
lilt

lilt
temperature at each part of the column changes at the same rate as the rest
of the column .

....

.,.,
Advantages of T P GC
lilt
....
The advantages ofT P GC can be illustrated by comparison of a

.,. chromatograms eluted at isothermal and temperature programed conditions.

Sample is a mixture of components with a very broad range of boiling
points. Elution starts with an oven temperature below the lowest boiling
....,
....
....
point of the components, and linearly increasing to the temperature

.,
.,. 71

....

".,
.
.,.
11.
1l1li ~ '! .!
"I: ~ ~
I! .¥f =
 ....
..

@II

..
....

72 ......
......
corresponding to the higher boiling point of the mixture.

At low oven temperature, early peaks, representing volatile components

with low boiling points, emerge as sharp symmetrical peaks, which depicts
higher column efficiency. With the increase of the temperature compounds
with higher boiling will start to evaporate and enter the mobile phase
moving as compact bands at the end of the column.

The main advantage of T P GC is the possibility to accommodate the

....
separation of samples with wide range ofboiling points. Secondly, and .­

most importantly, one achieves a shortening of the analytical time. .­

.­
Isothennal elution of a mixture, oven temperature 95 0 C; injector 220 0 C; .­
detector temperature 230 0 C.
.­
Same sample mixture eluted under programmed temperature:
.­
Initial value 95 0 C, Initial time 0 min, Rate 10 0 C / min
Final value 115 0 C, Final time 0 min
.­
WIll
is demonstrated in Figure 44.
.­
The figure above shows isothennal (a) and temperature
programmed (b) elution of a sample of heptane ( b. p. 110 0 C ), octane ( b.p.

130 0 C), and decane (b.p. 160 0 C); instmment HP 5890, column
Chromososrb W 5% DSCM.

T P GC requires a sophisticated control of the oven temperature,

......
WIll

gadget to perfonn the required temperature changes, and feed back

infonnation on temperature at each moment and every point of the controlled
volume.
Temperature programming allows the proper selection of
temperature resulting in total separation of all mixtures' components, and at
the same time sharply decreasing the analysis time.

72
..,

.,
.,

.,
.,

.,
.,

..,­

.,
.,
As one can notice in the figure bellow the elution time for the
73

.,
.,
last peak is significantly shorter than in isothennal elution. Shortening the
analytical time has a great economical effect.

.,
.,
Choosing proper values for the parameters of TPGC

.,
.,
Column length

In T P GC instruments, the length of the column is chosen considering the

.,
., required resolution. Packed columns are usually 2 - 3 meters long. Longer
columns are not beneficial in temperature progrannned instruments because

.,

.,
.,

they requires bigger ovens and imposes a limit on the heating rate of the
column.

.,
., Initial temperature

.,
., The initial temperature is chosen considering the lower boiling point of the
components. Initial temperature is nonnally lower than the lowest boiling

.,.,
point of the mixture. The lower initial temperature has negligible effect upon
the resolution of the higher boilers since these solutes are essentially frozen
and thus unaffected by the early parts of the temperature program.
.,., Rate of temperature increase

.,., The selection of the rate oftemperature increase is a compromise between

.,., resolution and analysis speed. The heating rate provides the same function in
temperature progrannned analysis as the operating temperature does in

.,., isothennal analysis .

.,
., .

.,
.,

.,

II!
II!
II!
 t!!!!"
~
t!!'
t!!'
f!!'
f!J!I'
fI/IJ
f!J!I'
74 fI/IJ
At lower rates, analysis time will be too long for the high boilers and band
ft!II
width deterioration will take place. At high rates, severe loss of resolution
f!!I'
will occur.
f!J!I'
Final temperature f!!I'
f!!I'
The final temperature is the highest temperature which the oven reaches by f!!I'
this temperature rate. It has to be selected being closely below the boiling
point of the highest boiling point of the mixture. One has to note that even at
.,.,
f!!I'

temperature lower than the boiling point of a compound, its molecules will be

distributed between gaseous phase and stationary phase, and will propagate .,
through the column.

.,.,
fI!!I/J
Qualitative evaluations in T P GC
.,
Identification of the peaks in T P CG is performed the same way
as the identification of the peaks in isothermal elution. If the retention times
......
of two compounds eluted at the same conditions (all parameters of the
temperature program should be similar) are equivalent, this is accepted as a
proof that these compounds are identical.

Retention time is defined also as the time which particular compound spends
..
inside the column during the separation process.

Identification of the compound by external standard at TP conditions.

If a standard has the same retention time as one ofthe peaks ofthe T P GC

chromatogram, compound corresponding to this peak is accepted as identical

74
 75
to the standard.
One has to be aware that all parameters in the temperature program
for the elution of the sample have to be identical to the parameter of the
program for elution of the standards.

One ramp temperature program

Most popular temperature program include a linear increase of the

temperature from a lower to the higher point. This type ofprogram can also
include an initial platform of constant temperature and/or one platform of
constant temperature at the final temperature. See Figure 45.

Change of the oven temperature during the process of separation ­

temperature program - is created by the operator of the GC instrument,

considering the properties of the sample.

To build a program one has to enter the values for the following parameters:

Initial temperature This is the temperature of the column at the beginning of

the analysis. One has to make an intelligent choice based on the boiling

points of the mixture. Initial temperature is similar or lower than the lowest

boiling point.

Initial time This parameter determines the time, during which the

thermostat has to hold the initial temperature constant. This time is

programmed in minutes. If one enters 0.5 that means that for 30 seconds the

oven temperature will be kept at the initial value. The length of this period

depends on the number of eluants with lower boiling points, and how close

those boiling points are. If one enters 0 min (from the Fig. 45-case a) or c)

75
 f!!!'"
f!/IIJ
f!!"
f!!"
f!!IIIJ

76
then the temperature of the oven will start to increases at once.
.....
f!!IIIJ
f!!1I'

f!!!II'
"

f!!!II'
Rate Rate is the increase of the oven temperature in degrees l

f!!IIJ
Celsius per minute. There are limits of the maximum rate increase. Some ,
f!!IIJ
instruments tolerate increase of up to 30 0 Cor 40 0 C maximum.
fI!!IIJ
Final value Final value is the maximum oven temperature reached in fI!!IIJ
this separation process. fI!!IIJ,
fI!!IIJ.
Final time Final time is the time period during which the oven is kept at fI!!IIJ
temperature of the final value.
,.,.,
fI!!IIJ

Run time is the time taken to complete the separation of the sample mixture.

It is a sum of the initial time, time needed to reach final value, and the final

time. During this time the process of separation and analysis is completed.

The temperature program will automatically return the oven to the initial

temperature ready for the next run or experiment.

Total time includes the run time, and the time needed to cool the oven to the

initial value, and to achieve equilibrium. The instrument cools down for an

uncontrolled period. After this period has elapsed the conditions are adjusted

to those at the beginning of the run.

76
,

..
~

.,.,
~ 77

...... Two ramps temperature program

.... More complicated mixtures can be eluted under a two ramp temperature

......
program. In this case the oven temperature is increased in two consecutive steps. To
create two ramps program one has to enter values for the following parameters:

First ramp:

...... 1.
2.
Initial value - starting temperature of the oven;
Initial time - time during which the initial temperature is constant;

......
3. Rate - the increase of the column temperature;
4. Final value - maximum temperature reached with this rate;
5. Final time the time period of oven temperature at final value .

......

Second ramp:

6. Rate A - new rate of increase of column temperature;

......
7. Final value A - is the maximum temperature reached at this rate of increase of
oven temperature;
8. Final time A - is the time period during which the oven temperature is held at
it final value .

...... After the final time is finished the run is competed. Uncontrolled cooling and
equilibrium times are needed to bring the oven temperature at the initial conditions,

...... ready to start the separation of another sample .

Example: Set-points for two ramps program:

....
..
Initial value
Initial time
80 0 C
0.5 min

..
..
..

..
..

.•

77

78
Rate 20 0 C / min. Rate A 15 0 C
Final value 140 0 C. Final value 180 0 C
Final time 1 min Final time 2 min

See Figure 47 for the graphical presentation of two ramps program

Three ramps temperature program

Temperature programming in modem instruments is designed to

give the technologist the opportunity to create even more sophisticated
temperature programs - with three ramps. The goal is to achieve full
separation at shortest analytical time. In this case oven temperature is
increased in three steps with different rates and different final times. To
create a three ramps program one has to enter values for the following
parameters:

First ramp:

1. Initial value - starting temperature ofthe oven;

2. Initial time - time during which the initial temperature is constant;
3. Rate - rate of increase of the column temperature;
4. Final value - maximum oven temperature reached with this rate;
5. Final time - time period with oven temperature at final value.

Second ramp:
6. Rate A - new rate of increase of column temperature;
7. Final value A - maximum temperature reached at this rate of increase of
oven temperature;
8. Final time A - time period during which the oven temperature is held at it=s
final value.

78
III!

III!

III!

III!

III!

lit

-e 79

~
~ Third ramp
~
9. Rate B- new rate of temperature increase of the oven;
II! 10. Final value B - maximum temperature (or final temperature) achieved with
III! this rate;
II! 11. Final time B - period of time at which the oven is held at temperature final
II! value B.
12. After the final time is completed, the run is stopped .. Uncontrolled cooling
III! and equilibrium times are needed to bring the oven temperature at the initial
III! conditions, ready to start the separation of another sample. The cooling is
~ initiated automatically and no specific command is needed.
III!
Example: Example set-points for three ramps program:
III!
III!
First ramp Initial value 80 0 C
~
Initial time 0.5 min
~
Rate 20 0 C / min,
~ Final value 140 °C.
~ Final time 1 min
~ Second ramp
II! Rate A
II! Final value A 180 0 C
II! Final time A 2 min.
~ Third Ramp
II! RateB °c
Final value B 200 C
-e Final time B 1 min
0

-e See Figure 50.

-e There are several additional requirements imposed to Temperature
-e
-e 79
-e
ae
ae
-e
e
-e
e
 ...
@II

..
..
..

@II

..
..
..

.....
80
Programmed GC instrument which are not required for an instrument not
furnished with temperature programming. These requirements are:

Reqnirements for TP GC
......
1. Stationary phase
......
As has been observed it was a complicated business to choose a proper
liquid phase for a separation at isothermal conditions and this task is even ......
harder for T P GC. Liquid phase must remain a liquid even at the
maximum operating temperature, and not evaporate. the vaporization of the ......
liquid phase is often referred to as bleeding. Ifthe column bleeds it
produces noise. A common limit to the maximum vapor pressure is 10 -6 g
liquid per milliliter of carrier gas. Use oflow liquid loading and narrow
......
columns helps avoid bleeding.
.....
Solidification ofthe liquid phase is also unadvisable. Freezing point or low
viscosity determines the minimum operating temperature of liquid phase. .....
2. Purity of the carrier gas.

For isothermal GC it is not very essential to use high purity inert gas. On

the contrary, Temperature Programmed GC requires high purity carrier gas.

Therefore, special oxygen traps and moister traps are very common

accessories to each of the modem GC instruments.

Separate column, detector, and the injection port thermostats.

It is very essential for a temperature programmed GC instrument to

80
~

•­
•­

•...

e
e
81
have separate heating systems for the column, the detector and the injection
e port.
e The thermostat that controls the temperature of the oven in T P GC
e is a relatively complex and advanced apparatus. It includes a computer chip,
which collects the information, controls the temperature and executes the
e program. There are also thermocouples, embedded in the walls of the oven,
e which measure temperature in different parts of the oven, and sent this
e information to the controller and to the electrical circuit to switch the heater
e ON and OF. Separate electrical circuit controls the upper temperature limit
e of the column. There is also a heater indicator, that allows the technologist
e to monitor constantly the temperature of the oven.
e
e 2. Wide boiling points
e
e Majority of the samples of natural products, such as crude oil extract, plant
e and animal oil extracts, food products and others, have very wide rage of
e boiling point. In many cases the boiling points of the new products are even
not known. Isothermal elution of such type of samples will require low
e starting temperature to accommodate volatile components. Viscose
e components will need very long elution times several hours.
e
e Temperature Programmed GC can analyze complex samples much faster.
e Low initial temperature will favor the elution of volatile components.
e Majority of the components with higher boiling points will stay frozenln
Ie the beginning of the column. The increase of the oven temperature will
Ie facilitate the elution of the higher boilers. As the oven temperature
Ie increases furthelmore viscose samples' components will enter the gaseous
Ie phase, and propagate through the column. Final oven temperature is
e
e 81
e
e
e
e
e
e
~
 ""
~

,.
".

,.
".

(IIJ
(IIJ
82 (IIJ
determined upon the stability of the liquid phase. (IIJ
(IIJ
3. Sample injection
~
Isothermal gas chromatography requires fast injection of the sample into ~
the injection port. The temperature of the injection port is settled above the
fI!IJ
boiling point of the higher boiler in the sample mixture. The reason for this t!'
is that all the sample components evaporate fast and enter the column as a t!'
compact band or as a "plug". This is critical if one wants to obtain better IJf!IJ
separation and sharp and distinguished peaks. ~
Fast injection of the sample is not critical for Temperature ~
Programmed gas chromatography. Volatile compounds will evaporate and ~
enter the column at once, but more viscose components will enter the f!!IJ
column slowly. They can even solidify at the beginning of the column, if
IJf!IJ
oven temperature, at this moment, is significantly lower than their boiling
~
temperature. With the increase of the oven temperature components with
appropriate boiling point will start to vaporize and enter the mobile phase. ~
(I!fJ
One can summarize that the main requirements for Temperature f!!J
Programmed Gas Chromatography in the following manner: f!!'
~
l. Separate thermostats ~
2. Computer-programmer
3. Low mass oven
4. Suitable liquid phase
5. Pure carrier gas

82
 83
Summary

Particle size of the solid support. Column efficiency is improved by the

use of small unifonn particles for solid support.

Flow rate. For maximum efficiency the column must be operated at the
optimum flow rate. This is expressed on the van Deemster plot. The minimum
HETP determines the optimum linear gas velocity.

Carrier gas. Carrier gas should have high molecular weight to achieve
optimal efficiency.

Liquid phase. Liquid phase should have low viscosity, low vapor pressure
with good solubility for the sample components.

Pressure. Optimal efficiency is obtained at low inlet to outlet ratio. The

majority of the gas chromatographs work at an outlet pressure of one
atmosphere.

Resolution can usually be improved by lowering the column temperature.

Column temperature is a very important parameter in the operation of a GC

instrument. It deserves special attention and will be discussed in detail in the
chapter on temperature programming. At this point one can summarize that
both the partition coefficient, and the diffusion coefficients are temperature
sensitive, therefore the entire process of separation is temperature dependent.
Decrease of the column temperature increases the resolution between the
peaks. Increase of the column temperature shortens the elution time.

83