GAS CHROMATOGRAPHY

GC is useful for compounds which are naturally volatile or can be converted into a volatile form. GC has been a widely used method due to its high resolution, low detection limits, accuracy and high resolution limits, and short analytical time. Retention of a compound is determined by its vapour pressure and volatility which, in turn, depends on its interaction with stationary phase.

Two types of stationary phases are commonly used in GC are solid absorbent (Solid Gas Chromatography [GSD]), and liquid coated on solid support (Gas-Liquid Chromatography [GLC]). In GSC same material (usually silica, alumina, or activated carbon) act as both the stationary phase and support phase. GLC uses liquid phases such as polymers, hydrocarbons, fluorocarbons, liquid crystals, and molten organic salts to coat the solid support materials.

 Calcin

diatomaceous earth graded into appropriate size ranges used as a stationary phase because it is stable inorganic substance.  Components of a typical GC system consists five components: a gas cylinder as a mobile phase source, a sample injector, a column, a detector, and a computer for data acquisition.  The mobile phase used in GC usually inert gas such as nitrogen, helium, hydrogen, argon, carbon dioxide, nitrous oxide and ammonia.

 The

carrier gas should be of high purity, and the flow must be tightly controlled to ensure optimum column efficiency and reproducibility of test results.  Samples are introduced in to GC using hypodermic syringe or automated samplers.  Each injection port is heated to very high temperatures to derivatize it into more volatile form by derivatization reactions like silylation, alkylation, and acylation.

Silylation is most common technique which replaces active hydrogen on compounds with alkylsilyl group. This substitution results in a more volatile form that is less polar and more thermally stable. Retention of compounds in a GC column can be adjusted by changing temperature. The column temperature affects volatility of compounds and thus the degree of their interaction with the stationary phase.

High performance Liquid Chromatography

 HPLC

uses small rigid support and special mechanical pumps producing high pressure to pass the mobile phase through the column.  HPLC column may be used several times without regeneration.  The resolution achieved by HPLC is superior than any other forms of liquid chromatography.  There are five commonly used separation techniques in liquid chromatography.  They include adsorption, partition, ion exchange, affinity and size exclusion.

 Each

is characterized by a unique combination of stationary phase and mobile phase. ______________________________________