A Research on Numerous Techniques and Alternatives of Spectroscopy and Chromatographic Evaluation | Original Article

ABSTRACT:

Chromatographic methods utilize partitioning a sample between two phases, one of which is the stationary phase and the other one is the mobile phase. Equipment for liquid chromatography includes a column, where separation takes places, pump, which generates a mobile phase flow, and detector, where the separated compounds are detected. Chromatography is a method which separates molecules based on differences in their structures, namely in size, in presence of charged, polar, and non-polar groups or moieties interacting specifically with an affinity column. After separation in a chromatographic column, particles are eluted to the detector and they are detected as chromatographic peaks. There are several physico-chemical principles of separation which can be employed in liquid chromatography: ion exchange chromatography, which separates molecules according to their charges; hydrophobic and reversed phase chromatography, which separate according to hydrophobicity of molecules; gel-permeation chromatography, which separates molecules according to their size; and affinity chromatography, which is based on specific interactions between the solute and molecules attached to the column.