Post on 19-Jan-2016
Chromatography
Chromatography
• Separation technique that relies on the affinity of components of a mixture to:-– A Mobile Phase– A Stationary Phase
• All types of chromatography rely on this– Paper chromatography– thin layer chromatography, – high performance liquid chromatography, – gas chromatography
Chromatography
• Mobile Phase– This is the phase that is a carrier for the
components as they travel through the stationary phase
• Stationary Phase– This is the phase that is stationary!!
Types of Chromatography
• Paper Chromatography• Thin layer chromatography (TLC)• High Performance Liquid Chromatography
(HPLC)• Gas Chromatography (GC)
there are more......................
Paper Chromatography
• Basic technique• Stationary Phase – Paper• Mobile Phase – Usually water• Sample preparation• Good for separating water soluble inks• Limited types of mobile phases can be used
therefore limit to what can be separated• Can be slower • Detection – naked eye or UV light
Paper chromatography.rv
Thin Layer Chromatography
• Improvement on paper technique• Stationary Phase – silica or alumina based
– Acidic, basic, neutral• Mobile Phase – solvents (varied polarities)• Good for:-
– Monitoring progress of a reaction– Identification of components in a mixture– Purity determination– Conditions for column separation techniques
• Semi-quantative • Can be quicker than paper – phases• Detection – UV light, chemical spray, iodine tank
Rf Values (retention factor)
• Distance travelled by component
• Distance travelled by mobile phase
• Specific to each plate• Comparable• Each component on
plate has unique Rf value
High Performance Liquid Chromatography
• Mobile phase – solvents/water/mixes• Stationary phases – silica based/many types for certain
applications• Sample preparation• Precise injection volumes• Composition/flow of mobile phase controlled• Oven temperature• Widely used in analytical laboratories• Quantify components – non volatile mixtures• Retention times
Gas Chromatography
• Mobile phase – unreactive gas (helium, nitrogen)• Stationary phases – polymer or liquid on an inert
solid support• Sample preparation• Precise injection volumes• Temperature/gas flow control• Widely used in analytical laboratories• Quantify components – volatile mixtures• Retention times
Columns
Injector
Autosampler
Detector
Data collection software
Column oven
AutosamplerData collection software
Column oven Keypad
Injector
Detector
Applications in Pharma Industry
Chromatogram
The Future• Predictive software• Screening• Saving resources