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Chromatography
 Definition

It is the method of separation of components in sample being analyzed by bringing two

immiscible phases ; one mobile and one stationary in contact with each other

Carrier
(Mobile
Integrator /
Phase) Recorder /
Computer
Sample

Detector
Column

 Types of Chromatography

Chromatography

Gas Chromatography High Performance Liquid Chrmatography

Gas Liquid Chromatography Partition Chromatography

Adsorption Chromatography

Gas Solid Chromatography

Ion Exchange Chromatography

Gel Permeation Chromatography

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Overview of Gas Chromatograph
 Sample Injection
Stage 1

Column Detector
Supply of carrier GasSample Injection System Column Detector
Integrator / Recorder / Computer
Oven

Carrier

Hydrogen Sample In Packed

TCD
Sample
Loop
Sample Out

Detector
FIDColumn
Stage 2
Nitrogen Capillary

Carrier

Helium FPD

Sample In Sample
Loop

Sample Out
CHROMATOGRAM

RT1 RT2

Conc
 Peak Lumping
CHROMATOGRAM ANALYSIS

PEAK DETECTION Method

Forced Gating Slope Detection External

Standard

Noise Calculation Normalized

Standard

Peak Width Indirect External

Standard

Peak Lumping

Tangent Skim

Base Line Definition

Suspend End

End = crest plus

Area Rejection
 Peak Lump

Tangent Skim

B
2

A
D

Peak End = Valley Peak End = Base

Area Measured Area Measured

Peak End
Peak End
 Suspend End
Chromatography Principles

Chromatography is the method in which two immiscible phases are brought in contact with
each other with one phase being stationary and the other mobile. The sample mixture introduced
Various
into the mobile phase undergoes a series of interactions many baselines determined
times between stationary and
mobile phase as it is carried through the stationary phase by the mobile phase. Interactions
exploit the differences in the physical and chemical properties of the sample. These differences
govern the rate of migration of individual components under the influence of mobile phase
moving through the column containing the stationary phase. Separated components emerge in
order of increasing interaction with the stationary phase. The least retarded component elutes
first and the most retained component elutes last

The mobile phase can be Gas or Liquid where as the stationary phase can only be solid or
liquid.

Chromatography employing gaseous mobile phase is known as GAS CHROMATOGRAPHY

It can be broadly classified into two types
 GLC GAS LIQID CHROMATOGRAPHY when the stationary phase is a liquid
 GSC GAS SOLID CHROMATOGRAPHY when the stationary phase is a solid
Suspend End
Chromatography employing On mobile phase is known as HPLC Suspend
a liquid i.e. HIGHEnd Off
PERFORMANCE LIQUID CHROMATOGRAPHY
It can be broadly classified into
 Partition Chromatography
 End = Crest Plus
Adsorption Chromatography
 Ion Exchange Chromatography
 Gel Permeation Chromatography
These can at times employ liquid mobile phase at very high pressure of about 5000 psi and is
hence also known as High Pressure Liquid Chromatography
Crest

Gas Chromatograph
Basically a GC consists of six parts
 Supply of carrier gas
 Sample Injection System
 Column
 Detector Specified End
 Integrator / Recorder
 Oven

Carrier Gas
The carrier gas provides the mobile phase in the system and is generally supplied by high
pressure cylinders. The carrier gas should be of high purity free any organic impurities which
can give erroneous reading
Generally Nitrogen or Hydrogen or helium are the most popular choice of carrier gasses.
End = Crest +

Sample Injection System

 The sample must be introduced as a vapour in the smallest possible volume and in minimum
amount of time without decomposition or fractionation occurring. Both quantity and manner of
introduction should be reproducible with high degree of precision. Large volume of sample
spreads over a greater length and overloads the column. Smaller sample gives a better peak
shape
A calibrated sample loop is provided to ensure reproducible sample injection volume

Column
Two types of columns are available
 Packed
 Capillary

Detector
Once the components elutes from the column it is detected in the Detector
The following types of detectors are available in an On-line Chromatograph
 Flame Ionization Detector ( FID )
 Thermal Conductivity Detector ( TCD )
 Flame Photometric Detector ( FPD )
The output of a detector plotted on a recorder is known as a chromatogram of concentration
versus time
The temperature of detector should be high enough to prevent condensation of sample vapour

Integrator
The Integrator provides method to analyze the chromatogram and identify the component and
enables the plot of the chromatogram

Oven
Oven provides constant temperature to the system and houses the column.

Chromatogram
The chromatogram which is a plot of concentration versus time is a series of peaks and valleys
of varying area and sizes. Each peak corresponds to a particular component and the time at
which it occurs is known as the Retention Time ( RT ) of the component. The Retention time
depends upon the characteristics of the component and the carrier gas flow rate. The size of peak
depends upon the concentration of the component.
RT1 RT2

Conc
 Online Gas Chromatograph

The on-line GC includes an electronic housing , an insulated air heated oven compartment a
purge air Panel and a pressure control panel

GC Controller

Time

Purge Air Electronic

Panel Pressure
controller

Oven

The Oven Compartment houses an isothermal oven which contains analytical columns ,
detectors and sample valves
The GCC contains card cage with PCB’s , the Temperature Controller and one or more of the
following detector amplifiers FID , TCD , FPD. The front panel has Liquid Crystal Display
( LCD) keypad and controller board

Below the GCC are the Purge Air panel and the Pressure control Panel
The Purge Air panel contains the heater air and purge air gauges and regulators
The Pressure Control panel contains the Electronic Pressure Controller ( EPC ) or a group of
mechanical regulators and gauges

The whole system is certified to Zone 1 Group IIC T3 hazardous area classification as per
CENELEC

In the event of purge air failure alarm is issued to alert the operator

Sample Flow Description

Sample Valves provide control and measurement of samples

The valves are air actuated by an electrical solenoid valve
An operational cycle consists if 3 modes
 Sample Purge
 Sample Inject
 Backflush

Sample Purge
Here the sample flows through the sample loop to vent

Sample Inject
The valve switched and the carrier gas sweeps the sample loop through the
sample/backflush valve , through Column 1 , through Column 2 and finally to detector
where it is converted to electrical signal
Column 1 performs preliminary separation to remove heavier non-measured components
which are then backflushed to vent
Column 2 separates the components to be measured which elutes sequentially
The selector valve determines which components will be measured. The sample output
from column 2 goes through column 3 to the detector for measurement . When the valve
switches the components not being measured is flushed through to selector vent

Backflush
Here the carrier gas backflushes the Column 1 to vent
 Flame Ionization Detector

It consists of a polarized jet , a collector and a chamber in which hydrogen fuel and sample are
burnt in air . A current whose magnitude is proportional to the concentration of the sample flows
between the jet and the collector.
A thermocouple senses the flame temperature and in event of flame failure the ignitor assembly
will re-ignite the flame. The output of detector is sent for signal processing

FID is used for ppm level measurement and is responsive to sample that can burn in hydrogen to
produce a signal
It cannot be used for measurement of sample of Oxygen Nitrogen CO2 etc
For %age level analysis a TCD is generally preferred

Thermal Conductivity Detector

TCD cell consists of single metal block with a pair of matched filaments RT1 & RT2 forming a
part of Wheatstone Bridge. The reference carrier gas flows over RT1 while the sample and
carrier gas from column flows over RT2 . When only carrier gas flows in both filaments they
have the same temperature and the bridge is balanced . As sample flows there is change in
filament temperature which changes the electrical resistance of the filament and causes an
imbalance of the bridge which produces an output
 Chromatogram Analysis

Method Tables

A Method table is a program used to control an analyser during an analysis and contains all
instructions and data required to identify each component of the sample and produce a
meaningful analysis report

In order for analysis to be accurate the GC must be calibrated first

Calibration provides a response factor which converts the individual components peak area or
height into an actual component concentration value . This response factor is generated by one
of the following three calibration method
 External Standard
 Normalized Standard
 Indirect External Standard

External Standard
Here the controller calibrates the analyzer against an external calibration standard of
precisely known component concentration ( CC ). Each component must be contained in
the calibration stream so that detector response can be determined for each component
independent of the other components. The system performs an analysis of the calibration
stream obtaining corrected values of peak area ( A ) and peak height ( H ) of each
component . The Response Factor ( RF ) is then calculated as
RF = CC/A for peak area analysis
or
RF = CC/H for peak height analysis

Normalized
The controller calibrates the analyzer against the external standard then calculates the
sum of all peaks and determines the percent contribution of each component.

The Response Factor ( RF ) is then calculated as

RFi = (Atot/Ai)/Cci
where i is for individual component.

Indirect External Standard

This mode is used when components cannot be individually calibrated . It uses a
previously developed set of response factor to calculate the relationship between the key
component and the other components
Here we provide the concentration of a specific component in a calibration stream and
the Relative Response Factor of each component to be measured . The system performs
analysis of the calibration stream to obtain corrected peak area value for the calibration
component (AC) . A K-factor is then calculated by the formula

K= (CCc/AC)(1/RRFc)
where subscript “c” is for calibration component
Component Concentration is the obtained as
CCi = (Ai)(K)(RRFi)
where “i” is for individual component

Calibrating the key components updates only the K-factor while other components are
updated through the relationship of various components
 Peak Detection

The GC measures peak in two ways

 Forced Gating
 Slope Detection

Forced Gating
This method is used when the components elution time is known . Here the time is
specified during which only the peak area is measured

Slope Detection
It is a method of determining peaks starting and ending point. The controller continously
scans for changes which indicate the start of peak , the peak maxima and the end of peak
It utilises the following function
 Noise Calculation
 Peak Width
 Peak Lumping
 Tangent Skim
 Baseline Definition
 Suspend End
 End=crest plus
 Area Rejection

Noise Calculation
It used to determine the noise level during the period of inactivity. This helps in deciding
when the slope of chromatogram indicates a peak change and not a random fluctuation
due to noise
Peak Width
It is used to calculate the minimum slope of the start of peak
Peak Lumping
It allows grouping of consecutive peaks into one peak
Tangent Skim
If a small peak overrides a large peak this is used to identify the two peaks . It searches
for the tangent termination point which begins in the valley point
Baseline Definition
If the controller is unable to befine whether the end of a peak is a valley point or baseline
the controller will give wrong result . This enables to define the ambiguity as baseline or
a valley
Suspend End
It permits suspension of automatic baseline definition
End= crest+
It forces the end of the peak to be crest of the peak plus the time specified in the code
Area Rejection
It is used to specify the peaks of area which are to be rejected from the analysis
 GC Utilities

1 SAMPLE CONDITIONING MIN NORM. MAXI

 FLOW RATE LIQUID 50
SAMPLE CC/MIN
 FLOW RATE VAPOUR 50 100
SAMPLE CC/MIN
 INLET TEMPERATURE BUBBLE POINT
LIQUID SAMPLE or 150 OC
 INLET TEMPERATURE 150 OC
VAPOUR SAMPLE
 INLET PRESSURE LIQUID 30
SAMPLE ( BAR )
 INLET PRESSURE VAPOUR 30 PSI
SAMPLE
 OUTLET PRESSURE ATMOS. 30
LIQUID SAMPLE (BAR)
 OUTLET PRESSURE ATMOS. 30 PSI
VAPOUR SAMPLE ( BAR )

2 UTILITIES REQUIREMENT PRESS. FLOW TEMP

INSTRUMENT AIR 60 - 100 PSIG 4.5 - 9.5 SCFM AMBIENT

VALVE AIR REQUIREMENT
PURGE INSTRUMENT AIR
FID PURIFIER
CARRIER GAS 6 BAR 150 - 200 CC/MIN AMBIENT
BURNER FUEL ( HYDROGEN 40 CC/MIN
FOR FID )
HYDROGEN PURITY 99.9995%
BURNER AIR 400 CC/MIN