UNIT-IV

GAS CHROMATOGRAPHY

Points to be covered in this topic

- INTRODUCTION
- THEORY
- INSTRUMENTATION
- DERIVATIZATION
- ADVANTAGES
- DISADVANTAGES
- APPLICATIONS

INTRODUCTION

Gas Chromatography

A.T. James and P. Martin first time used the gas chromatography technique in 1952 for separating long cha
Gases and vaporisable substances can also be separated by gas chromatography based on differential ad
In gas chromatography, gas is used as the mobile phase and solid or liquid is used as the stationary phase
When the stationary phase is solid, it is known as Gas Solid Chromatography (GSc) and when the stationa
Gas Solid In gas chromatography, a moving gas phase is passed over a stationary sorbent to separate the

This technique is similar to that of liquid-liquid chromatography with the only exception that in the former a m

Inlet
Column
Power Switch
GAS CHROMATOGRAPHY
Detector
Control Panel (Touchscreen)
Oven
PRINCIPLE

In gas chromatography, the substance to be analysed is partitioned between the mobile and stationary pha
During the separation, the sample is vaporised and carried through the column by the mobile gas phase (i.e
The different components get separated based on their vapour pressure and affinities for the stationary pha

The affinity of a component towards the stationary phase is termed as distribution constant (Kc), which is a

Where.
Kc = [A]s / [A]m

[A]s = concentration of component A in the stationary phase

[A]m = concentration of component A in the mobile phase.

Movement of different components through the column is controlled by the distribution constant (Kc), thus t

Schematic Representation of the Chromatographic Process

(Direction of mobile-phase flow)

(Concentration of solute in mobile phase)
(Concentration of solute in stationary phase)
(Detector Chromatogram)

THEORY

The separation of compounds in a mixture is based on different polarities in a direct (interaction with station
The gas chromatography column consists of solid support that is covered with a high-boiling liquid in a thin
In the example, compound "X" has a higher affinity towards the stationary phase compared to compound "O
Compound "O" elutes before compound "X" because it displays a lower boiling point and a weaker interacti

Stationary phase (liquid phase)

Solid Support
Compound mixture
Carrier Gas (mobile phase)

Factors that influence the outcome in the GC

Vapor Pressure of the Compound

The higher the boiling point of the compound is, the lower the vapor pressure will be.
Thus, the compound with higher boiling point migrates slower through the column resulting in a longer reten
Low boiling solvents like diethyl ether, dichloromethane, ethyl acetate, methanol, etc. are usually used to di

Polarity of the Compound and the Polarity of the Column

The stronger the interaction of the compound with the stationary phase is going to be increasing the retenti
Very polar compounds (i.e., alcohols, amines) often cause tailing on polar columns (or older columns).
A variety of column polarities is available for the separation of compounds using gas chromatography.

INSTRUMENTATION

The following components make up the instrumentation of gas chromatography:

1. Carrier gas maintained at a high pressure and delivered at a rapid and reproducible rate,
2. Sample injector,
3. Separation columns,
4. Detectors,
5. Thermostated chambers for regulating the temperature of column and detectors,
6. Amplifier and recorder system.

Carrier Gas
Carrier gases like hydrogen, helium, nitrogen, and air are widely used.
Hydrogen is most advantageous but also dangerous to use.
Helium is inert and thermally conductive but expensive.
Nitrogen is cheap but reduces sensitivity.
Air is used when oxygen is useful to the detector or separation.

Carrier Gas selection criteria include inertness, purity, suitability for detector, column performance, safety, a
Sample Injector
Used to introduce sample reproducibly and vaporize it rapidly.
Liquid samples injected through self-sealing rubber septum using hypodermic syringe.
Solid samples dissolved in volatile liquids or directly injected if liquefiable.
Gas samples introduced using special gas sampling valves.

Separation Columns
Made of glass or metal tubing, diameter around 4.8 mm.
Lengths range from a few cm to 100 m.
Types: Packed columns, open tubular (capillary) columns, support-coated open tubular, wall coated open t

Detectors
Common detectors include Flame Ionisation Detector (FID) and Thermal Conductivity Detector (TCD).
Detector properties: high sensitivity, low volume, rapid and linear response, stability, and flow/temperature

Katharometer uses thermal conductivity changes to detect compounds.

Flame Ionisation Detector relies on change in flame conductivity caused by burning compounds.
Thermal Conductivity Detector measures heat loss of filament due to thermal conductivity of surrounding ga
Thermionic Emission Detector uses fuel-poor hydrogen plasma and low temperature flame, selective for nit
Electron Capture Detector detects electron affinity, sensitive to halogenated compounds.

DERIVATIZATION
Derivatization improves separation or detection.
Two types: Pre-column (modifies sample to be more volatile/thermostable) and Post-column (improves det
Pre-column derivatization examples: converting carboxylic acids, sugars, phenols, alcohols to less polar co
Pretreatment of solid support improves stationary phase film uniformity, reduces peak tailing.

TEMPERATURE PROGRAMMING
Column temperature is increased gradually during analysis.
Improves peak shapes, resolution, and reduces analysis time.

ADVANTAGES
Rapid analysis, high efficiency, sensitive detectors, small sample volume, non-destructive, high quantitative
DISADVANTAGES
Limited to volatile and thermally stable samples, not suited for preparative chromatography, structural analy

APPLICATIONS
Qualitative analysis, purity check, impurity detection, quantitative analysis (direct comparison, calibration cu

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Introduction
Used to separate complex chemical mixtures.
Previously called High Pressure Liquid Chromatography.
Offers better resolution, less time, more accuracy, precision, and sensitivity compared to classical liquid ch
Uses high pressure to pump mobile phase through packed column.

Principle
Separation based on partition behavior between mobile phase and stationary phase packed with small part
Four separation methods: adsorption, partition, ion exchange, exclusion.

Theory
HPLC uses high pressure pump unlike gravity-fed classical chromatography.
Mobile phase is polar or non-polar solvent mixture.
Compounds interact differently with stationary phase leading to separation.
Weak interaction leads to early elution, strong interaction to late elution.

Instrumentation
Includes solvent reservoir with degassing, pump system, sample injection, columns, detectors, recorder, da

Solvent reservoir holds mobile phase, degassing removes gases to avoid pump/detector issues.

Pumps types include syringe pump, reciprocating pump, pneumatic pump.

Sample injection systems include septum injector, stop-flow septum-less injection, micro-volume sampling
Columns include guard column (protects analytical column) and analytical column (main separation column

Detectors include UV detector (common), fluorescence detector, refractive index detector, multipurpose de

Strip chart recorder records detector signals over time.

Data handling system automates analysis.

Advantages
Simple, fast, sensitive, high resolution, accurate, precise, small solvent amount, reproducible, useful for qua

Disadvantages
Requires UV-absorbing chromophore, some compounds need derivatization, stationary phase pH stability

Applications
Stability studies, bioassays, dosage form design, cosmetic industry quality control, isolation of natural produ