Thin layer chromatography as a tool for the separation of biomolecules

Thin-Layer Chromatography as a tool for the separation of biomolecules

Chromatography encompasses a diverse but related group of methods that permits the separation , isolation , identification and quantification of components in a mixture . Thin layer chromatography is a modern analytical separation method with extensive versatility , much of which is already utilized but there is vast potential for future development into areas where research is just beginning (Wall , 2005 ,

.2 . Thin layer chromatography is a sub-division of liquid chromatography , in which the mobile phase is a liquid and the stationary phase is situated

br as a thin layer on the surface of a flat plate . In TLC , the sample is applied as a small spot or streak to the origin of a thin sorbent layer supported on a glass , plastic or a metal plate . Of the three , glass is proved to be most popular , although aluminium or plastic is more advantageous in the sense that they are flexible and can easily be cut into any size with minimum disruption to the sorbent layer (Wall , 2005 br

.1 ) Various sorbents have been used including silica gel , cellulose alumina , polyamides , ion exchangers , and chemically bonded polar and non-polar phases . The mobile phase moves through the stationary phase through capilliary action (Fried and Sherma , 1999 ,

.1 , 9

Chromatographic techniques are powerful tools for the analysis of biomolecules . Various types of chromatography are useful , especially for analysing protein and nucleic acid . However , most methods tend to be complex due to complex nature of these molecules , thin layer , and high performance liquid chromatography are generally employed in the analysis of proteins and nucleic acids in biological materials such as food (Kenkel , 2003 ,

.476 ) Proteins vary in a number of their physical and chemical properties as a result of their amino acid sequences . The amino acid residues to the polypeptide backbone may be positively or negatively charged , neutral or polar or neutral or hydrophobic . In addition , the polypeptides are folded in definite secondary and tertiary structures to create a unique size , shape and distribution of residues on the surface of the protein . By exploiting the differences in properties between proteins in the mixture , a rationale technique to separate them can be designed (Luo , Andrade Caldwell , 1998 , 97-98

Thin-Layer Chromatography (TLC ) allows separation of PAHs with minimal sample clean up . Multiple samples and standards can be chromatographed in parallel on a single plate . Silica gel and alumina layers have used for the adsorption TLC separation of aromatic hydrocarbons , silica gels or cellulose layers impregnated with dymethylformamide for normal phase partition separations and acetylated cellulose layers for normal or reverse phase partition separations . Mixed alumina acetylated cellulose layers have also been used . However , these conventional TLC systems have not been completely successful in separate many complex PAH mixtures and HPTLC has been increasingly used rather than TLC (Rathore , 1993 br

.51 . High performance Thin Layer Chromatography (HPLTC ) has improved the sensitivity , speed , and separating power of TLC while preserving its traditional advantage of methodological simplicity , static...